• Reproductive and Developmental Biology
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• 제30권3호
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• pp.181-187
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• 2006

최근 돼지의 장기를 사람에게 이식하는 이종간 장기 이식에 관한 연구가 급속히 발전되고 있다. 그러나 돼지의 장기를 이식할 경우 가장 큰 문제점 중의 하나는 돼지 genome 내에 존재하는 내인성 레트로바이러스(porcine endogenous retrovirus; PERV)가 인간에게 그대로 전이될 수 있다는 것이다. 이에 대한 대안으로 최근 활발히 연구되고 있는 RNA 간섭을 통한 PERV RNA의 발현을 최대한 억제하는 방법이 제안되고 있는데, RNA 간섭(RNA interference)은 double-standard RNA(dsRNA)가 상보적인 표적 mRNA를 분해하여 결과적으로 표적 단백질의 발현을 특이적으로 억제하는 현상을 의미한다. 본 연구에서는 PERV에 대한 RNA 간섭 현상을 일으키는 shRNA 유전자를 레트로바이러스 벡터를 이용하여 돼지세포에 RNA)가 상보적인 표적 mRNA를 분해하여 결과적으로 표적 단백질의 발현을 특이적으로 억제하는 현상을 의미하다. 도입한 후 PERV의 발현율 감소 여부를 조사하였다. 그 결과, gag-pol 유전자와 env 유전자 발현은 각각 대조군 세포의 4%와 10% 정도로 억제되었다. 한편, virus 입자의 생산에서 gag-pol 유전자는 대조군 세포에 비해 300배 이상 억제되었으며, env 유전자에서는 20만 배 이상 억제되었다. 이상의 결과를 미루어 볼 때 형질 전환 돼지를 이용한 이종 장기 이식에 있어서 RNA 간섭 현상을 이용한 PERV의 발현을 억제하는 시도는 생물학적안전성을 크게 증가시킬 수 있을 것으로 사료된다.

• 한국어병학회지
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• 제26권3호
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• pp.149-161
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• 2013

바이러스성 출혈성 패혈증을 일으키는 VHSV는 국내의 넙치양식에 심각한 피해를 주고 있다. 본 연구에서는 우리나라에서 분리한 8종의 VHSV 분리주들의 glycoprotein (G)의 염기서열을 분석하여 Korean VHSV 분리주들과 기보고된 일본 및 중국의 분리주들을 계통발생학적으로 비교하여 G-protein의 8 부위(G34, C528, A755, T834, T951, T1147, T1221, T1336)에서 국내 VHSV 분리주들의 특이적인 서열을 확인하였고, 전 세계적으로 분리된 VHSV의 분리주들과의 비교결과 8 부위 중 3 부위 (A755, T834, T1221)의 국내VHSV 분리주 특이적 염기서열들을 확인하였다. 또한 중국 유래 VHSV 분리주의 특이적 염기서열 11 부위 (C498, C545, C548, C642, G648, T678, A738, C804, C834, G851, C1139)와 일본 유래 VHSV 분리주의 특이적 염기서열 1부위 (C179)를 확인하였다. 이상의 연구결과는 국내 특이적인 염기서열을 확보하여, 구역화된 수산생물 질병의 예찰, 모니터링 및 질병 관리에 유용하게 사용될 것으로 사료된다.

• Genes and Genomics
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• 제40권11호
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• pp.1169-1180
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• 2018

Cave shrimps from the genera Typhlatya, Stygiocaris and Typhlopatsa (TST complex) comprises twenty cave-adapted taxa, which mainly occur in the anchialine environment. Anchialine habitats may undergo drastic environmental fluctuations, including spatial and temporal changes in salinity, temperature, and dissolved oxygen content. Previous studies of crustaceans from anchialine caves suggest that they have possessed morphological, behavioral, and physiological adaptations to cope with the extreme conditions, similar to other cave-dwelling crustaceans. However, the genetic basis has not been thoroughly explored in crustaceans from anchialine habitats, which can experience hypoxic regimes. To test whether the TST shrimp-complex hypoxia adaptations matched adaptive evolution of mitochondrial OXPHOS genes. The 13 OXPHOS genes from mitochondrial genomes of 98 shrimps and 1 outgroup were examined. For each of these genes was investigated and compared to orthologous sequences using both gene (i.e. branch-site and Datamonkey) and protein (i.e. TreeSAAP) level approaches. Positive selection was detected in 11 of the 13 candidate genes, and the radical amino acid changes sites scattered throughout the entire TST complex phylogeny. Additionally, a series of parallel/convergent amino acid substitutions were identified in mitochondrial OXPHOS genes of TST complex shrimps, which reflect functional convergence or similar genetic mechanisms of cave adaptation. The extensive occurrence of positive selection is suggestive of their essential role in adaptation to hypoxic anchialine environment, and further implying that TST complex shrimps might have acquired a finely capacity for energy metabolism. These results provided some new insights into the genetic basis of anchialine hypoxia adaptation.

• Biomolecules & Therapeutics
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• 제27권4호
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• pp.414-422
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• 2019

There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

• Animal Bioscience
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• 제34권1호
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• pp.93-101
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• 2021

Objective: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. Methods: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. Results: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. Conclusion: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

• 식품과학과 산업
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• 제55권3호
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• pp.276-283
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• 2022

비타민D3는 칼슘 대사를 조절하며 뼈의 건강을 담당하는 중요한 호르몬이지만 다양한 면역세포에 작용하여 면역반응을 조절할 수 있다는 것이 알려지면서 자가면역질환, 알레르기질환, 감염 등 여러 질병과의 상관관계에 대한 관심이 증가되었다. 비타민D3의 활성 형태인 1,25(OH)2D3는 면역세포에 의해 전환될 수 있으며 핵-호르몬 수용체인 VDR과 결합하여 세포 내 다양한 유전자 발현을 조절함으로써, 세포의 형질과 기능을 바꿀 수 있다. 특히, 최근 다양한 연구를 통해 과도한 면역 반응을 조절하는 Treg 세포의 분화를 증가시키고 그 기능을 향상시키는 기전을 통해 염증성 사이토카인을 감소시키며, 외부 병원균을 제거할 수 있는 AMP의 생산을 유도함으로써 자가면역질환, 알레르기, 감염성 질환 등 다양한 질환에 긍정적 역할을 할 수 있음이 확인되었다. 특히, 최근 COVID-19 에 의한 글로벌 팬데믹을 경험하면서, 중증 질환 및 사망에 이르는 환자들과 비타민D의 상관관계 연구, 다양한 나라들에서 시행된 비타민D 치료의 임상결과들을 통해, 비타민D 보충 기반의 면역조절의 전략이 유리할 수 있음을 알 수 있었다. 전세계 현대인들의 비타민D 결핍은 최근 50년 간 지속적으로 증가하고 있으며, 자외선 노출로 합성할 수 있는 비타민D의 양이 충분하지 않은 부분을 식품 또는 보충제의 형태로 섭취하는 것이 여러 자가면역질환과 알레르기, 감염성 질병을 예방하는데 도움이 될 것이다.

• Journal of Microbiology and Biotechnology
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• 제33권2호
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• pp.188-194
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• 2023

Microbacterium elymi KUDC0405^T was isolated from the rhizosphere of Elymus tsukushiensis from the Dokdo Islands. The KUDC0405^T strain was Gram-stain-positive, non-spore forming, non-motile, and facultatively anaerobic bacteria. Strain KUDC0405^T was a rod-shaped bacterium with size dimensions of 0.3-0.4 × 0.7-0.8 ㎛. Based on 16S rRNA gene sequences, KUDC0405^T was most closely related to Microbacterium bovistercoris NEAU-LLE^T (97.8%) and Microbacterium pseudoresistens CC-5209^T (97.6%). The dDDH (digital DNA-DNA hybridization) values between KUDC0405^T and M. bovistercoris NEAU-LLE^T and M. pseudoresistens CC-5209^T were below 17.3% and 17.5%, respectively. The ANI (average nucleotide identity) values among strains KUDC0405^T, M. bovistercoris NEAU-LLE^T, and M. pseudoresistens CC-5209^T were 86.6% and 80.7%, respectively. The AAI (average amino acid identity) values were 64.66% and 64.97%, respectively, between KUDC0405^T and its closest related type strains. The genome contained 3,596 CDCs, three rRNAs, 46 tRNAs, and three non-coding RNAs (ncRNAs). The genomic DNA GC content was 70.4%. The polar lipids included diphosphatydilglycerol, glycolipid, phosphatydilglycerol, and unknown phospholipid, and the major fatty acids were anteiso-C_17:0 and iso-C_16:0. Strain KUDC0405^T contained MK-12 as the major menaquinone. Based on genotypic, phylogenetic, and phenotypic properties, strain KUDC0405^T should be considered a novel species within the genus Microbacterium, for which we propose the name M. elymi sp. nov., and the type strain as KUDC0405^T (=KCTC 49411^T, =CGMCC1.18472^T).

• The Korean Journal of Physiology and Pharmacology
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• 제27권4호
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• pp.417-426
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• 2023

The TRPM4 gene encodes a Ca²⁺-activated monovalent cation channel called transient receptor potential melastatin 4 (TRPM4) that is expressed in various tissues. Dysregulation or abnormal expression of TRPM4 has been linked to a range of diseases. We introduced the hemagglutinin (HA) tag into the extracellular S6 loop of TRPM4, resulting in an HA-tagged version called TRPM4-HA. This TRPM4-HA was developed to investigate the purification, localization, and function of TRPM4 in different physiological and pathological conditions. TRPM4-HA was successfully expressed in the intact cell membrane and exhibited similar electrophysiological properties, such as the current-voltage relationship, rapid desensitization, and current size, compared to the wild-type TRPM4. The presence of the TRPM4 inhibitor 9-phenanthrol did not affect these properties. Furthermore, a wound-healing assay showed that TRPM4-HA induced cell proliferation and migration, similar to the native TRPM4. Co-expression of protein tyrosine phosphatase, non-receptor type 6 (PTPN6 or SHP1) with TRPM4-HA led to the translocation of TRPM4-HA to the cytosol. To investigate the interaction between PTPN6 and tyrosine residues of TRPM4 in enhancing channel activity, we generated four mutants in which tyrosine (Y) residues were substituted with phenylalanine (F) at the N-terminus of TRPM4. The YF mutants displayed properties and functions similar to TRPM4-HA, except for the Y256F mutant, which showed resistance to 9-phenanthrol, suggesting that Y256 may be involved in the binding site for 9-phenanthrol. Overall, the creation of HA-tagged TRPM4 provides researchers with a valuable tool to study the role of TRPM4 in different conditions and its potential interactions with other proteins, such as PTPN6.

• IMMUNE NETWORK
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• 제23권6호
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• pp.45.1-45.22
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• 2023

Interstitial lung disease (ILD) involves persistent inflammation and fibrosis, leading to respiratory failure and even death. Adult tissue-derived mesenchymal stem cells (MSCs) show potential in ILD therapeutics but obtaining an adequate quantity of cells for drug application is difficult. Daewoong Pharmaceutical's MSCs (DW-MSCs) derived from embryonic stem cells sustain a high proliferative capacity following long-term culture and expansion. The aim of this study was to investigate the therapeutic potential of DW-MSCs in experimental mouse models of ILD. DW-MSCs were expanded up to 12 passages for in vivo application in bleomycin-induced pulmonary fibrosis and collagen-induced connective tissue disease-ILD mouse models. We assessed lung inflammation and fibrosis, lung tissue immune cells, fibrosis-related gene/protein expression, apoptosis and mitochondrial function of alveolar epithelial cells, and mitochondrial transfer ability. Intravenous administration of DWMSCs consistently improved lung fibrosis and reduced inflammatory and fibrotic markers expression in both models across various disease stages. The therapeutic effect of DW-MSCs was comparable to that following daily oral administration of nintedanib or pirfenidone. Mechanistically, DW-MSCs exhibited immunomodulatory effects by reducing the number of B cells during the early phase and increasing the ratio of Tregs to Th17 cells during the late phase of bleomycin-induced pulmonary fibrosis. Furthermore, DW-MSCs exhibited anti-apoptotic effects, increased cell viability, and improved mitochondrial respiration in alveolar epithelial cells by transferring their mitochondria to alveolar epithelial cells. Our findings indicate the strong potential of DW-MSCs in the treatment of ILD owing to their high efficacy and immunomodulatory and anti-apoptotic effects.

• 농업과학연구
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• 제49권4호
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• pp.1015-1023
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• 2022

Production of high-quality horses is important to make the horse industry grow. Grazing during the growing period can be an important factor affecting the production of high-quality horses. The objective of this study was to determine the effects of grazing on growing horses by analyzing their blood components and intestinal microbiota. Twelve growing horses for evaluating blood components and ten growing horses for evaluating intestinal microbiota were raised for about seven months and separated by two treatments: grazing vs. stable. Complete blood count, blood chemistry, and creatine kinase levels were analyzed as blood components and a 16s rRNA gene sequence analysis was performed to analyze intestinal microbiota. Calcium ions tended to be lower in the group with grazing treatment. Alkaline phosphatase and creatine kinase tended to be higher in the group with grazing treatment. These results indicate that grazing can provide horses with more exercise than staying in stables. At the phylum level, Firmicutes/Bacteroidetes ratios in grazing and stable groups were 4.2 and 6.5, respectively. Because various studies have reported that a. high Firmicutes/Bacteroidetes ratio indicates obesity, the method of raising horses might affect their physical ability. At the species level, rates of Clostridium butyricum in grazing and stable groups were 3.2% and 13.1%, respectively. Some strains of C. butyricum can cause several diseases such as botulism. These results indicate that grazing can positively affect growing horses in terms of blood components and intestinal microbiota. Moreover, grazing can be helpful to make growing horses healthy through proper exercise.