• Molecules and Cells
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• 제39권7호
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• pp.543-549
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• 2016

MicroRNAs (miRNAs) have been reported to be involved in many neurodegenerative diseases. The present study focused on the role of hsa-miR-144-3p in one of the neuro-degenerative diseases, Parkinson's disease (PD). Our study showed a remarkable down-regulation of miR-144-3p expression in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-treated SH-SY5Y cells. MiR-144-3p was then overexpressed and silenced in human SH-SY5Y cells by miRNA-mimics and miRNA-inhibitor transfections, respectively. Furthermore, ${\beta}$-amyloid precursor protein (APP) was identified as a target gene of miR-144-3p via a luciferase reporter assay. We found that miR-144-3p overexpression significantly inhibited the protein expression of APP. Since mitochondrial dysfunction has been shown to be one of the major pathological events in PD, we also focused on the role of miR-144-3p and APP in regulating mitochondrial functions. Our study demonstrated that up-regulation of miR-144-3p increased expression of the key genes involved in maintaining mitochondrial function, including peroxisome proliferator-activated receptor ${\gamma}$ coactivator-$1{\alpha}$(PGC-$1{\alpha}$), nuclear respiratory factor 1 (NRF-1) and mitochondrial transcription factor A (TFAM). Moreover, there was also a significant increase in cellular ATP, cell viability and the relative copy number of mtDNA in the presence of miR-144-3p overexpression. In contrast, miR-144-3p silencing showed opposite effects. We also found that APP overexpression significantly decreased ATP level, cell viability, the relative copy number of mtDNA and the expression of these three genes, which reversed the effects of miR-144-3p overexpression. Taken together, these results show that miR-144-3p plays an important role in maintaining mitochondrial function, and its target gene APP is also involved in this process.

• 비교문화연구
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• 제20권
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• pp.5-27
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• 2010

This paper studies the relation between canonical/variegated babble (CB/VB) and early speech in an infant acquiring Mandarin Chinese from 9 to 17 months. The infant was audio-and video-taped in her home almost every week. The data analyzed here come from 1,621 utterances extracted from 23 sessions ranging from 30 minutes to one hour, from age 00:09;07 to 01:05;27. The data was digitized, and segments from 23 sessions were transcribed in narrow IPA and coded for analysis. Babble was coded from age 00:09;07 to 01:00;00, and words were coded from 01:00;00 to 01:05;27, proto-words appeared at 11 months, and some babble was still present after 01:10;00. 3821 segments were counted in CB/VB utterances, plus the segments found in 899 word tokens. The data transcription was completed and checked by the author and was rechecked by two other researchers who majored in Chinese phonetics in order to ensure the reliability, we reached an agreement of 95.65%. Mandarin Chinese is phonetically very rich in consonants, especially affricates: it has aspirated and unaspirated stops in labial, alveolar, and velar places of articulation; affricates and fricatives in alveolar, retroflex, and palatal places; /f/; labial, alveolar, and velar nasals; a lateral;[h]; and labiovelar and palatal glides. In the child's pre-speech phonetic repertoire, 7 different consonants and 10 vowels were transcribed at 00:09;07. By 00:10;16, the number of phones was more than doubled (17 consonants, 25 vowels), but the rate of increase slowed after 11 months of age. The phones from babbling remained active throughout the child's early and subsequent speech. The rank order of the occurrence of the major class types for both CB and early speech was: stops, approximants, nasals, affricates, fricatives and lateral. As expected, unaspirated stops outnumbered aspirated stops, and front stops and nasals were more frequent than back sounds in both types of utterances. The fact that affricates outnumbered fricatives in the child's late babble indicates the pre-speech influence of the ambient language. The analysis of the data also showed that: 1) the phonetic characteristics of CB/VB and early meaningful speech are extremely similar. The similarities of CB/VB and speech prove that the two are deeply related; 2) The infant has demonstrated similar preferences for certain types of sounds in the two stages; 3) The infant's babbling was patterned at segmental level, and this regularity was similarly evident in the early speech of children. The three types being coronal plus front vowel; labial plus central and dorsal plus back vowel exhibited much overlap in the phonetic forms of CB/ VB and early speech. So the child's CB/ VB at this stage already shared the basic architecture, composition and representation of early speech. The evidence of similarity between CB/VB and early speech leaves no doubt that phones present in CB/VB are indeed precursors to early speech.

• 한국육종학회지
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• 제40권4호
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• pp.387-393
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• 2008

Ds 삽입변이체로 부터 5,400개의 FST를 분석한 결과, intragenic FST가 48.1%로 2,597개, intergenic FST가 25.6%로 1,383개였으며 hot spot을 포함한 origin insertional sequence는 1,350개로 25%로 나타났다. Intragenic FST로서 선발된 2,597개를 이용하여 Ds와의 유전자형을 분석한 결과 53.6%인 1,393개가 heterozygous 혹은 homozygous 계통으로 나타났으며 이들에 대한 염색체상의 분포도는 3번 염색체에서 422 계통으로 가장 많은 분포도를 보여주었고 다른 염색체는 56 계통에서 157 계통 범위 내에 포함되었다. 1차적으로 유전자형 분석이 끝난 1,393개의 유전자들 중에서 expressed protein 등 알려지지 않은 것은 40.6%로 566개였으며 TIGR DB에서 염기서열의 유사성 검색을 통해 유전자의 명칭이 알려진 것은 59.4%인 827개로 나타났다.

• Molecules and Cells
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• 제41권12호
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• pp.1008-1015
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• 2018

$I{\kappa}B$, a cytoplasmic inhibitor of nuclear factor-${\kappa}B$ ($NF-{\kappa}B$), is reportedly degraded via the proteasome. However, we recently found that long-term incubation with proteasome inhibitors (PIs) such as PS-341 or MG132 induces $I{\kappa}B{\alpha}$ degradation via an alternative pathway, lysosome, which results in $NF-{\kappa}B$ activation and confers resistance to PI-induced lung cancer cell death. To enhance the anti-cancer efficacy of PIs, elucidation of the regulatory mechanism of PI-induced $I{\kappa}B{\alpha}$ degradation is necessary. Here, we demonstrated that PI up-regulates nuclear factor (erythroid-derived 2)-like 2 (Nrf2) via both de novo protein synthesis and Kelch-like ECH-associated protein 1 (KEAP1) degradation, which is responsible for $I{\kappa}B{\alpha}$ degradation via macroautophagy activation. PIs increased the protein level of light chain 3B (LC3B, macroautophagy marker), but not lysosome-associated membrane protein 2a (Lamp2a, the receptor for chaperone-mediated autophagy) in NCI-H157 and A549 lung cancer cells. Pretreatment with macroautophagy inhibitor or knock-down of LC3B blocked PI-induced $I{\kappa}B{\alpha}$ degradation. PIs up-regulated Nrf2 by increasing its transcription and mediating degradation of KEAP1 (cytoplasmic inhibitor of Nrf2). Overexpression of dominant-negative Nrf2, which lacks an N-terminal transactivating domain, or knock-down of Nrf2 suppressed PI-induced LC3B protein expression and subsequent $I{\kappa}B{\alpha}$ degradation. Thus, blocking of the Nrf2 pathway enhanced PI-induced cell death. These findings suggest that Nrf2-driven induction of LC3B plays an essential role in PI-induced activation of the $I{\kappa}B$/$NF-{\kappa}B$ pathway, which attenuates the anti-tumor efficacy of PIs.

• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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• pp.257-264
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• 2019

Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

• Journal of Plant Biotechnology
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• 제46권3호
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• pp.180-188
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• 2019

옥신은 식물의 생장과 발달 과정에서 중요한 조절자로서 기능한다. 옥신 신호전달 과정은 3개의 주요 옥신 반응 전사인자인 Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), 그리고 small auxin up RNA (SAUR) 유전자에 의해 조절된다. 특히, Aux/IAA는 옥신 신호에 반응하여 빠르게 축적되는 수명이 짧은 핵 단백질이다. 이 실험에서 우리는 현사시 나무(Populus alba ${\times}$ P. glandulosa)로 부터 PagAux/IAA1 유전자를 분리하고 발현 특성을 분석하였다. PagAux/IAA1 cDNA는 4개의 보존된 도메인과 2개의 nuclear localization sequence (NLS)을 포함한 200개의 아미노산을 암호화하고 있다. Southern blot 분석으로 현사시나무 genome에 PagAux/IAA1 유전자가 single copy로 존재하는 것을 확인하였다. PagAux/IAA1 유전자는 잎과 꽃에서 특이적으로 발현되었다. 그리고 PagAux/IAA1 유전자는 현탁배양세포의 생장 과정에서 초기 지수생장기에 발현되었다. PagAux/IAA1 유전자의 발현을 분석한 결과, 건조와 염 스트레스 및 식물호르몬인 ABA 처리에 의해 발현이 감소된 반면 저온 스트레스, 형성층의 세포 분열 과정 그리고 식물호르몬인 GA와 JA 처리에서 발현이 증가하였다. 따라서 PagAux/IAA1 유전자가 현사시나무에서 저온 스트레스 반응뿐 아니라 생장 과정에 관여할 것으로 판단된다.

• 한국응용과학기술학회지
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• 제38권5호
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• pp.1292-1301
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• 2021

본 연구는 섬애약쑥(Artemisia argyi H.)의 항가려움증 활성 가능성을 확인하기 위한 연구로써 가려움증 관련인자 등을 측정하였다. 섬애약쑥은 열수로 추출(Artemisia argyi H. distilled water extract 이하, AAD)하여 MTT assay로 세포독성을 측정하였고 항 가려움증 활성을 확인하기 위하여 IL-4와 IL-31 관련 전사인자 발현 및 단백질 생성을 측정하였으며, 히스타민의 발현을 측정하였다. 그 결과 25, 50, 100 ㎍/㎖의 농도에서는 유의한 세포독성이 나타나지 않는 것을 확인하였다. 가려움증 연관 유전인자인 IL4의 경우 25 ㎍/㎖의 농도에서 약 12%, 50 ㎍/㎖의 농도에서 약 26%, 100 ㎍/㎖의 농도에서 약 61%로 유의하게 감소하였으며, IL31의 경우 50 ㎍/㎖의 농도에서 약 33%, 100 ㎍/㎖의 농도에서 약 33%로 유의하게 감소하였다. 연관된 단백질 측정의 경우 각각 50 ㎍/㎖와 100 ㎍/㎖의 농도에서 IL-4는 약 34% 및 약 69%, IL-31은 약 36% 및 약 37% 유의하게 감소하였다. 이 결과는 ADD가 항가려움증을 위한 소재로써 가능성을 보여 소재 개발을 위한 기초자료로 제공될 수 있을 것으로 사료된다.

• 한국약용작물학회지
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• 제28권6호
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• pp.395-411
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• 2020

Background: This study investigated the anti-obesity effect of the flavonoid rich fraction (FRF) and its constituent, rutin obtained from the leaf of Morus alba L., on the lipid accumulation mechanism in 3T3-L1 adipocyte and C57BL/6 mouse models. Methods and Results: In Oil Red O staining, FRF (1,000 ㎍/㎖) treatments showed inhibition rate of 35.39% in lipid accumulation compared to that in the control. AdipoRedTM assay indicated that the triglyceride content in 3T3-L1 adipocytes treated with FRF (1,000 ㎍/㎖) was reduced to 23.22%, and free glycerol content was increased to 106.04% that of the control. FRF and its major constituent, rutin affected mRNA gene expression. Rutin contributed to the inhibition of Sterol regulatory element binding protein-1c (SREBP-1c) gene expression, and inhibited the transcription factors SREBP-1c, peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer binding protein α (C/EBPα), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). In addition, the effect of FRF administration on obesity development in C57BL/6 mice fed high-fat diet (HFD) was investigated. FRF suppressed weight gain, and reduced liver triglyceride and leptin secretion. FRF exerted potential anti-inflammatory effects by improving insulin resistance and adiponectin levels, and could thus be used to help counteract obesity. The mRNA expressions of PPAR-γ, FAS, ACC, and CPT-1 were determined in liver tissue. Quantitative real-time PCR analysis was also performed to evaluate the expression of IL-1β, IL-6, and TNF-α in epididymal adipose tissue. Compared to the control group, mice fed the HFD showed the up-regulation in PPAR-γ, FAS, IL-6, and TNF-α genes, and down-regulation in CPT1 gene expression. FRF treatement markedly reduced the expression of PPAR-γ, FAS, IL-6, and TNF-α compared to those in HFD control, whereas increased the expression level of CPT1. Conclusions: These results suggest that the FRF and its major active constituent, rutin, can be used as effective anti-obesity agents.

• Animal Bioscience
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• 제34권8호
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• pp.1392-1402
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• 2021

Objective: The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells. Methods: Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5×10⁵ cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (MyoD) expression and growth-related gene expression in JBP muscle cells, respectively. Results: We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (EGF+bFGF) than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of collagen type XXI alpha 1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells. Conclusion: Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.

• 한국미생물·생명공학회지
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• 제49권4호
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• pp.485-492
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• 2021

본 연구는 오크라 추출물의 미백 효과를 검증하여 화장품 소재로서 활용가능성을 확인하고자 하였다. 먼저, 오크라 열수 및 70% 에탄올 추출물의 미백효과를 tyrosinase의 효소 억제 활성으로 측정한 결과, 최종 농도인 1,000 ㎍/ml 농도에서 22.2%, 32.8%의 저해활성 효과를 보였다. 세포 차원에서 미백효과를 측정하기 위해 오크라 열수 및 에탄올 추출물의 세포 생존율을 melanoma cell (B16F10)에서 MTT assay법을 이용하여 측정하였다. 그 결과, 오크라 열수 및 70% 에탄올 추출물에서 100 ㎍/ml 농도에서 95% 이상의 생존율을 보였으며, 세포 독성이 나타나지 않은 농도 이하에서 멜라닌 생합성을 확인하기 위해 실험을 진행하였으며, 농도 의존적으로 멜라닌 합성을 저해하는 것을 확인하였다. 오크라 열수 및 70% 에탄올 추출물의 단백질 발현억제 효과를 5, 10, 50, 100 ㎍/ml의 농도에서 western blot으로 측정하였으며, 양성대조군으로 β-actin을 사용하였다. 그 결과, 오크라 열수 추출물은 100 ㎍/ml 농도에서 MITF, tyrosinase, TRP-1, TRP-2 인자들은 각각 88.1%, 24.8%, 62.2%, 42.9%의 효과를 나타내었다. 오크라 70% 에탄올 추출물은 100 ㎍/ml 농도에서 MITF, tyrosinase, TRP-1, TRP-2 인자들은 각각 65.3%, 58.3%, 66.2%, 65.3%의 효과를 나타내었다. 결론적으로 오크라 열수 및 70% 에탄올 추출물의 미백 효과가 검증되었으며, 기능성 화장품 소재로서 활용가능성을 확인하였다.