• 대한수의학회지
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• 제61권2호
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• pp.13.1-13.8
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• 2021

Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 10^5.7 50% tissue culture infectious dose (TCID₅₀)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

• Journal of Veterinary Science
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• 제19권6호
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• pp.735-743
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• 2018

We investigated the effect of transforming growth factor beta 1 ($TGF-{\beta}1$) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with $TGF-{\beta}1$ at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of $TGF-{\beta}1$. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by $TGF-{\beta}1$ stimulation was dose and time dependent. $TGF-{\beta}1$ was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in $TGF-{\beta}1$ treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the $TGF-{\beta}1$-treated scaffolds. Together, our results suggest that $TGF-{\beta}1$ has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

• Nutrition Research and Practice
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• 제17권6호
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• pp.1128-1142
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• 2023

BACKGROUND/OBJECTIVES: Inonotus obliquus has been used as antidiabetic herb around the world, especially in the Russian and Scandinavian countries. Diabetes is widely believed to be a key factor in Alzheimer's disease (AD), which is widely considered to be type III diabetes. To investigate whether I. obliquus can also ameliorate AD, it would be interesting to identify new clues for AD treatment. We tested the anti-AD effects of raw Inonotus obliquus polysaccharide (IOP) in a mouse model of AD (3×Tg-AD transgenic mice). MATERIALS/METHODS: SPF-grade 3×Tg-AD mice were randomly divided into three groups (Control, Metformin, and raw IOP groups, n = 5 per group). β-Amyloid deposition in the brain was analyzed using immunohistochemistry for AD characterization. Gene and protein expression of pertinent factors of the ubiquitin-proteasome system (UPS) was determined using real-time quantitative polymerase chain reaction and Western blotting. RESULTS: Raw IOP significantly reduced the accumulation of amyloid aggregates and facilitated UPS activity, resulting in a significant reduction in AD-related symptoms in an AD mouse model. The presence of raw IOP significantly enhanced the expression of ubiquitin, E1, and Parkin (E3) at both the mRNA and protein levels in the mouse hippocampus. The mRNA level of ubiquitin carboxyl-terminal hydrolase isozyme L1, a key factor involved in UPS activation, also increased by approximately 50%. CONCLUSIONS: Raw IOP could contribute to AD amelioration via the UPS pathway, which could be considered as a new potential strategy for AD treatment, although we could not exclude other mechanisms involved in counteracting AD processing.

• Clinical Nutrition Research
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• 제12권1호
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• pp.7-20
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• 2023

Creating a complex balance between dietary composition, circadian rhythm, and the hemostasis control of energy is important for managing diseases. Therefore, we aimed to determine the interaction between cryptochrome circadian clocks 1 polymorphism and energy-adjusted dietary inflammatory index (E-DII) on high-sensitivity C-reactive protein in women with central obesity. This cross-sectional study recruited 220 Iranian women aged 18-45 with central obesity. The 147-item semi-quantitative food frequency questionnaire was used to assess the dietary intakes, and the E-DII score was calculated. Anthropometric and biochemical measurements were determined. By polymerase chain response-restricted length polymorphism method, cryptochrome circadian clocks 1 polymorphism was assigned. Participants were categorized into three groups based on the E-DII score, then categorized according to cryptochrome circadian clocks 1 genotypes. The mean and standard deviation of age, BMI, and high-sensitivity C-reactive protein (hs-CRP) were 35.61 ± 9.57 years, 30.97 ± 4.16 kg/m², and 4.82 ± 5.16 mg/dL, respectively. The interaction of the CG genotype and E-DII score had a significant association with higher hs-CRP level compared to GG genotype as the reference group (β, 1.19; 95% CI, 0.11-2.27; p value, 0.03). There was a marginally significant association between the interaction of the CC genotype and the E-DII score with higher hs-CRP level compared to the GG genotype as the reference group (β, 0.85; 95% CI, -0.15 to 1.86; p value, 0.05). There is probably positive interaction between CG, CC genotypes of cryptochrome circadian clocks 1, and E-DII score on the high-sensitivity C-reactive protein level in women with central obesity.

• 생명과학회지
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• 제29권12호
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• pp.1305-1313
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• 2019

감초(Glycyrrhiza inflata)의 뿌리에서 분리된 Licochalcone (LC)은 항염증 및 항종양과 같은 많은 약리학적 효과를 가지고 있다. 현재까지 LCC는 구강암과 방광암에서 연구되었지만 폐암에서의 연구는 밝혀진 바 없다. 또한, 암에서 LCC에 의해 유도된 autophagy에 대한 연구는 없었다. 본 연구는 gefitinib-민감성 또는 내성을 갖는 폐암 세포에 대한 LCC의 효과 및 작용 메커니즘을 조사하기 위해 고안되었다. MTT 분석 데이터는 LCC가 비소세포폐암 세포주인 HCC827 (gefitinib-민감성) 및 HCC827GR (gefitinib-내성)에서 세포생존율을 유의하게 억제함을 보여주었다. 흥미롭게도, Annexin V/7-aminoactinomycin D 이중 염색 및 세포주기 분석에서 가장 높은 농도의 LCC 처리는 apoptosis를 유도하는 비율이 약 10%였다. LCC는 비소세포폐암 세포주에서 세포주기 G2/M 관련 단백질인 cyclin B1 및 cdc2의 발현을 감소시킴으로써 G2/M 정지를 야기하였다. LCC의 처리는 autophagy marker 단백질인 microtubule-associated protein 1 light chain 3 (LC3) 및 autophagy과정에 관여하는 단백질인 autophagy-related gene (Atg)5의 발현을 증가시킴으로써 autophagy를 유도하였다. 또한, LCC는 reactive oxygen species (ROS)의 생성을 증가시켰으며, ROS 억제제인 N-acetyl-L-cysteine (NAC)에 의해 세포생존율이 부분적으로 회복되었다. Western blotting 분석에서, NAC과 LCC의 동시처리에 의해 cdc2의 발현이 증가하고 LC3의 발현은 감소되었다. 이러한 결과는 LCC가 비소세포폐암에서 ROS-의존적 G2/M 정지 및 autophagy를 유도함으로써 항종양 효과에 기여할 수 있음을 나타낸다. 결론적으로, LCC 치료는 비소세포폐암에 대한 잠재적 치료제로서 유용할 수 있다.

• Genomics & Informatics
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• 제10권2호
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• pp.110-116
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• 2012

Recent several studies have shown that the genetic variation of SCN5A is related with atrioventricular conduction block (AVB); no study has yet been published in Koreans. Therefore, to determine the AVB-associated genetic variation in Korean patients, we investigated the genetic variation of SCN5A in Korean patients with AVB and compared with normal control subjects. We enrolled 113 patients with AVB and 80 normal controls with no cardiac symptoms. DNA was isolated from the peripheral blood, and all exons (exon 2-exon 28) except the untranslated region and exon-intron boundaries of the SCN5A gene were amplified by multiplex PCR and directly sequenced using an ABI PRISM 3100 Genetic Analyzer. When a variation was discovered in genomic DNA from AVB patients, we confirmed whether the same variation existed in the control genomic DNA. In the present study, a total of 7 genetic variations were detected in 113 AVB patients. Of the 7 variations, 5 (G87A-A29A, intervening sequence 9-3C>A, A1673G-H558R, G3578A-R1193Q, and T5457C-D1819D) have been reported in previous studies, and 2 (C48G-F16L and G3048A-T1016T) were novel variations that have not been reported. The 2 newly discovered variations were not found in the 80 normal controls. In addition, G298S, G514C, P1008S, G1406R, and D1595N, identified in other ethnic populations, were not detected in this study. We found 2 novel genetic variations in the SCN5A gene in Korean patients with AVB. However, further functional study might be needed.

• 한국어병학회지
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• 제20권2호
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• pp.109-117
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• 2007

본 연구에서는 국내 연구보고는 아직 없는 실정이나 양식터봇의 연쇄구균 병원체로 보고 된 바가 있는 제주 양식넙치에서 분리되는 γ 용혈성의 연쇄구균인 Streptococcus parauberis의 특성을 제시하고자 하였다. S. parauberis에 대한 분리균주의 탄소원을 기질로 이용하는 특성은 Annette and Collllins (1990)가 보고한 결과와 유사하였다.또한 분리된 균주들을 계통분류학적 분석 결과 Accesion number AY584477 하나의 그룹으로 묶였지만 S. iniae와는 다른 그룹으로 묶였다. 그러나 Collins가 Streptococcus iniae (X58306)와 S. parauberis (X89967)이 한 그룹으로 묶인다고 발표한 것과는 다른 결과를 나타냈다. 16S rRNA gene에 대한 염기서열 분석 결과는 S. parauberis (AY942572)와 98%이상의 상동성을 나타내어 S. parauberis로 동정하였다. S. parauberis는 S. uberis type Ⅱ로 분류되었으나 Williams and Collins (1990)에 의해서 S. parauberis라는 종명이 제시되었다. 이러한 결과로 볼 때 제주도 양식넙치의 연쇄구균증 병원체로 S. parauberis로 판단되나 이번 실험에서 감염실험이 수행되지 않아 앞으로 본 병원체에 대한 감염 실험과 병리학적인 연구가 필요하다고 사료되어진다.

• 한국어류학회지
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• 제18권4호
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• pp.283-292
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• 2006

넙치 (Paralichthys olivaceus) 신장에서 추출한 mRNA로부터 cDNA library를 제작하고 이를 이용하여 EST (Expressed sequence tag) 분석을 하였다. 넙치의 신장 cDNA library에서 무작위로 선별한 390개의 EST를 조사한 결과, 250개의 EST는 이미 밝혀진 유전자와 유사성이 있는 것으로 나타났으며, 140개의 EST는 새로운 유전자로 밝혀졌다. 유전자의 기능이 밝혀진 250개의 EST 중 14 (5.6%)개의 EST는 이미 알려진 넙치 EST와 상동성이 있는 유전자로 확인되었고, 198 (79.2%)개의 EST는 다른 생물에서 알려진 유전자와 상동성이 있는 것으로 나타났다. 그러나 38 (15.2%)개의 EST는 전혀 기능이 알려지지 않은 새로운 유전자로 밝혀졌다. EST 분석은 새로운 유전자 뿐만 아니라 기능적으로 중요한 유전자를 탐색하는데도 아주 강력한 연구 방법이다. 이에 따라 본 연구에서는 넙치 신장 EST 분석을 통해 C-, L-type lectin과 MHC class II-invariant chain like proteins (li) 같은 면역기능과 관련이 있는 여러 개의 유전자를 확인하였고, 이들 유전자들은 질병감염 후 유전자 발현에서 나타나는 차이를 분석하는데 이용되는 cDNA microarray 연구에 유용하게 사용될 것으로 보인다.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.787-801
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• 2001

The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely　identified the cDNA for a novel protein from cultured　PDL fibroblasts using subtraction hybridization between gingival fibroblasts and　PDL fibroblasts. The purpose　of this study was to determine the regulation by growth factors and cytokines on　PDLsl7 gene expression in　cultured human periodontal ligament cells and observe the immunohistochemical　localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

• Asian-Australasian Journal of Animal Sciences
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• 제31권8호
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• pp.1088-1097
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• 2018

Objective: It is commonly accepted that adiponectin binds to its two receptors to regulate fatty acid metabolism in adipocytes. To better understand their functions in the regulation of intramuscular adipogenesis in goats, we cloned the three genes (adiponectin [AdipoQ], adiponectin receptor 1 [AdipoR1], and AdipoR2) encoding these proteins and detected their mRNA distribution in different tissues. We also determined the role of AdipoQ in the adipogenic differentiation of goat skeletal muscle satellite cells (SMSCs). Methods: SMSCs were isolated using 1 mg/mL Pronase E from the longissimus dorsi muscles of 3-day-old female Nanjiang brown goats. Adipogenic differentiation was induced in satellite cells by transferring the cells to Dulbecco's modified Eagle's medium supplemented with an isobutylmethylxanthine, dexamethasone and insulin cocktail. The pEGFP-N1-AD plasmid was transfected into SMSCs using Lipofectamine 2000. Expression of adiponectin in tissues and SMSCs was detected by quantitative polymerase chain reaction and immunocytochemical staining. Results: The three genes were predominantly expressed in adipose and skeletal muscle tissues. According to fluorescence and immunocytochemical analyses, adiponectin protein expression was only observed in the cytoplasm, suggesting that adiponectin is localized to the cytoplasm of goat SMSCs. In SMSCs overexpressing the AdipoQ gene, adiponectin promoted SMSC differentiation into adipocytes and significantly (p<0.05) up-regulated expression of AdipoR2, acetyl-CoA carboxylase, fatty-acid synthase, and sterol regulatory element-binding protein-1, though expression of CCAAT/enhancer-binding $protein-{\alpha}$, peroxisome proliferator-activated receptor ${\gamma}$, and AdipoR1 did not change significantly. Conclusion: Adiponectin induced SMSC differentiation into adipocytes, indicating that adiponectin may promote intramuscular adipogenesis in goat SMSC.