• 제목/요약/키워드: Gene disruption

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Single-cell and spatial transcriptomics approaches of cardiovascular development and disease

  • Roth, Robert;Kim, Soochi;Kim, Jeesu;Rhee, Siyeon
    • BMB Reports
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    • 제53권8호
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    • pp.393-399
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    • 2020
  • Recent advancements in the resolution and throughput of single-cell analyses, including single-cell RNA sequencing (scRNA-seq), have achieved significant progress in biomedical research in the last decade. These techniques have been used to understand cellular heterogeneity by identifying many rare and novel cell types and characterizing subpopulations of cells that make up organs and tissues. Analysis across various datasets can elucidate temporal patterning in gene expression and developmental cues and is also employed to examine the response of cells to acute injury, damage, or disruption. Specifically, scRNA-seq and spatially resolved transcriptomics have been used to describe the identity of novel or rare cell subpopulations and transcriptional variations that are related to normal and pathological conditions in mammalian models and human tissues. These applications have critically contributed to advance basic cardiovascular research in the past decade by identifying novel cell types implicated in development and disease. In this review, we describe current scRNA-seq technologies and how current scRNA-seq and spatial transcriptomic (ST) techniques have advanced our understanding of cardiovascular development and disease.

Molecular Detection of Verotoxigenic Escherichia coli (VTEC) from Animal Feces for Screening VTEC-shedders

  • Kobayashi, Y.;Sato, M.;Taguchi, H.;Koike, S.;Nakatsuji, H.;Tanaka, K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권3호
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    • pp.423-427
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    • 2004
  • Seventy-six animals including cattle, sheep, horses, 6 species of zoo animals were employed for collection of fresh feces in order to detect verotoxigenic Esherichia coli (VTEC) by safe, quick and sensitive PCR-based molecular methods. Bacterial cell disruption with bead-beating followed by bacterial DNA purification with hydroxyapatide chromatography and gel filtration allowed DNA preparation from animal feces with high recovery and purity. A mountain goat was firstly shown by PCR and sequencing to shed verotoxin 2 gene (vt2) that was used to generate vt2 probe and second primer set for nested PCR to attempt more sensitive detection. Most sensitive nested PCR revealed that 45% of tested cattle and 47% of tested zoo animals were VTEC-positive, while least sensitive normal PCR detected VTEC from none of these animals except a mountain goat. Moderately sensitive detection by PCR in combination with hybridization suggested that the VTEC density varied between the VTEC-positive cattle.

NF-kB Activation by Disruption of Microtubule Array during Myogenesis of L6 Cells

  • Sangmyung Rhee;Lee, Kun-Ho;Hyockman Kwon
    • Animal cells and systems
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    • 제1권1호
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    • pp.63-69
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    • 1997
  • We have previously reported that NF-kB is involved in the regulation of nitric oxide synthase gene expression during differentiation of chick embryonic myoblasts. However, how NF-kB is timely activated during myogenesis remains elusive. One of the most prominent events in myogenesis is myoblast membrane fusion, which is accompanied with massive cytoskeletal reorganization. Here we show that the activity of NF-kB markedly increases in L6 rat myogenic cells that have just initiated morphological changes by treating nocodazole, a microtubule-disrupting agent. Furthermore, the induction of NF-kB activation was closely correlated with the myoblast fusion. In addition, a variety of agents that disrupt microtubules stimulated the myoblast fusion as well as the induction of NF-kB activation. In contrast, taxol, a microtubule-stabilizing agent, suppressed the induction of NF-kB activation and inhibited spontaneous differentiation of L6 cells as well. In addition, we found that the NF-KB in the cells consists of p50/p65 heterodimers. These results support the idea that reorganization of microtubule at early stages of differentiation plays a role as a signal for NF-KB activation during myogenesis.

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Crystal Structure of PAS factor from Vibrio vulnificus

  • Lee, Jun-Hyuck;Park, Seong-Hwan;Im, Young-Jun;Kim, Mun-Kyoung;Kang, Gil-Bu;Kim, Young-Ran;Rhee, Joon-Haeng;Eom, Soo-Hyun
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.72-72
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    • 2003
  • The PAS factor, whose gene has been cloned from V vulnifcus, is a protein secretion factor. Although the role of the PAS factor in Vibrio is still unknown, it may be involved with the bacterial protein secretion. The PAS factor is a 76 amino acid polypeptide, and its expression in E. coli cells makes the host cell membrane leaky, resulting in the excretion of periplasmic proteins into the culture medium. Highly expressed PAS factor is harmful to the cell, this may be due to a disruption of the membrane structure or function.

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Potential crosstalk of oxidative stress and immune response in poultry through phytochemicals - A review

  • Lee, M.T.;Lin, W.C.;Lee, T.T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권3호
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    • pp.309-319
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    • 2019
  • Phytochemicals which exist in various plants and fungi are non-nutritive compounds that exert numerous beneficial bioactive actions for animals. In recent years following the restriction of antibiotics, phytochemicals have been regarded as a primal selection when dealing with the challenges during the producing process in the poultry industry. The selected fast-growing broiler breed was more fragile when confronting the stressors in their growing environments. The disruption of oxidative balance that impairs the production performance in birds may somehow be linked to the immune system since oxidative stress and inflammatory damage are multi-stage processes. This review firstly discusses the individual influence of oxidative stress and inflammation on the poultry industry. Next, studies related to the application of phytochemicals or botanical compounds with the significance of their antioxidant and immunomodulatory abilities are reviewed. Furthermore, we bring up nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and nuclear factor kappa B ($NF-{\kappa}B$) for they are respectively the key transcription factors involved in oxidative stress and inflammation for elucidating the underlying signal transduction pathways. Finally, by the discussion about several reports using phytochemicals to regulate these transcription factors leading to the improvement of oxidative status, heme oxygenase-1 gene is found crucial for Nrf2-mediated $NF-{\kappa}B$ inhibition.

Effects of Antibiotics on the Uterine Microbial Community of Mice

  • Sang-Gyu Kim;Dae-Wi Kim;Hoon Jang
    • 한국발생생물학회지:발생과생식
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    • 제26권4호
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    • pp.145-153
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    • 2022
  • The gut microbiota is involved in the maintenance of physiological homeostasis and is now recognized as a regulator of many diseases. Although germ-free mouse models are the standard for microbiome studies, mice with antibiotic-induced sterile intestines are often chosen as a fast and inexpensive alternative. Pathophysiological changes in the gut microbiome have been demonstrated, but there are no reports so far on how such alterations affect the bacterial composition of the uterus. Here we examined changes in uterine microbiota as a result of gut microbiome disruption in an antibiotics-based sterile-uterus mouse model. Sterility was induced in 6-week-old female mice by administration of a combination of antibiotics, and amplicons of a bacteria marker gene (16S rRNA) were sequenced to decipher bacterial community structures in the uterus. At the phylum-level, Proteobacteria, Firmicutes, and Actinobacteria were found to be dominant, while Ralstonia, Escherichia, and Prauserella were the major genera. Quantitative comparisons of the microbial contents of an antibiotic-fed and a control group revealed that the treatment resulted in the reduction of bacterial population density. Although there was no significant difference in bacterial community structures between the two animal groups, β-diversity analysis showed a converged profile of uterus microbiotain the germ-free model. These findings suggest that the induction of sterility does not result in changes in the levels of specific taxa but in a reduction of individual variations in the mouse uterus microbiota, accompanied by a decrease in overall bacterial population density.

Myxococcus stipitatus DSM 14675의 melithiazol 생합성 유전자 분석 (Analysis of the Melithiazol Biosynthetic Gene Cluster in Myxococcus stipitatus DSM 14675)

  • 현혜숙;박수현;조경연
    • 한국미생물·생명공학회지
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    • 제44권3호
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    • pp.391-399
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    • 2016
  • Melithiazol은 점액세균 Melitangium lichenicola, Archangium gephyra, Myxococcus stipitatus에 의해 생산되는 항진균 물질이다. M. lichenicola의 melithiazol 생합성 유전자는 이미 알려져 있지만, A. gephyra와 M. stipitatus의 melithiazol 생합성 유전자들은 아직까지 밝혀져 있지 않다. 본 연구에서는 유전체 서열 분석과 돌연변이 분석을 통해 M. stipitatus DSM 14675 균주로부터 37.3 kb 크기의 melithiazol 생합성 유전자군을 발견하였다. 이 유전자군은 9개(MYSTI_04973−MYSTI_04965)의 유전자로 구성되어 있는데, 4개의 polyketide synthase 모듈과 3개의 non-ribosomal peptide synthase 모듈, 그리고 fumarylacetoacetate hydrolase, S-adenosylmethionine-dependent methyltransferase, nitrilase를 암호화하는 것으로 분석되었다. 플라스미드 삽입 돌연변이를 통해 MYSTI_04972 유전자 또는 MYSTI_04973를 불활성화시켰을 때 melithiazol 생산능이 상실되었다. MYSTI_04972부터 MYSTI_04965까지의 8개 유전자가 암호화하는 melithiazol 생합성 모듈의 구성은 M. lichenicola Me l46에서와 유사하였다. 하지만 첫 번째 유전자(MYSTI_04973)에 의해 암호화되는 로딩 모듈의 구성은 M. lichenicola Me l46과 달랐는데, 이러한 차이는 M. stipitatus 균주들이 어떻게 M. lichenicola Me l46과는 다른 구조의 melithiazol 유도체들을 생산하는지 설명해준다.

Anti-aging effects of Korean Red Ginseng (KRG) in differentiated embryo chondrocyte (DEC) knockout mice

  • Nam, Youn Hee;Jeong, Seo Yule;Kim, Yun Hee;Rodriguez, Isabel;Nuankaew, Wanlapa;Bhawal, Ujjal K.;Hong, Bin Na;Kang, Tong Ho
    • Journal of Ginseng Research
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    • 제45권1호
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    • pp.183-190
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    • 2021
  • Background: The circadian rhythm is the internal clock that controls sleep-wake cycles, metabolism, cognition, and several processes in the body, and its disruption has been associated with aging. The differentiated embryo chondrocyte (Dec) gene is related to circadian rhythm. To our knowledge, there are no reports of the relationship between dec gene expression and KRG effect. Therefore, we treated Dec gene knockout (KO) aging mice with KRG to study anti-aging related effects and possible mechanisms. Methods: We evaluated KRG and expression of Dec genes in an ototoxicity model. Dec genes expression in livers of aging mice was further analyzed. Then, we assessed the effects of DEC KO on hearing function in mice by ABR. Finally, we performed DNA microarray to identify KRG-related gene expression changes in mouse liver and assessed the results using KEGG analysis. Results: KRG decreased the expression of Dec genes in ototoxicity model, which may contribute to its anti-aging efficacy. Moreover, KRG suppressed Dec genes expression in liver of wild type indicating inhibition of senescence. ABR test indicated that KRG improved auditory function in aging mouse, demonstrating KRG efficacy on aging related diseases. Conclusion: Finally, in KEGG analysis of 238 genes that were activated and 158 that were inhibited by KRG in DEC KO mice, activated genes were involved in proliferation signaling, mineral absorption, and PPAR signaling whereas the inhibited genes were involved in arachidonic acid metabolism and peroxisomes. Our data indicate that inhibition of senescence-related Dec genes may explain the anti-aging efficacy of KRG.

Magnaporthe oryzae Effector AVR-Pii Helps to Establish Compatibility by Inhibition of the Rice NADP-Malic Enzyme Resulting in Disruption of Oxidative Burst and Host Innate Immunity

  • Singh, Raksha;Dangol, Sarmina;Chen, Yafei;Choi, Jihyun;Cho, Yoon-Seong;Lee, Jea-Eun;Choi, Mi-Ok;Jwa, Nam-Soo
    • Molecules and Cells
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    • 제39권5호
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    • pp.426-438
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    • 2016
  • Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

흰쥐 치수에서 Er:YAG laser에 의한 노출손상에 따른 조직학적 변화 및 CGRP 단백질의 발현 (A HISTOLOGICAL CHANGES AND CGRP EXPRESSION AFTER EXPOSURE INJURY BY ER:YAG LASER IN DENTAL PULP OF RAT)

  • 양재호;박종태;김규탁;김상봉;이난영;이상호;김흥중
    • 대한소아치과학회지
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    • 제35권1호
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    • pp.11-17
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    • 2008
  • 본 연구는 Er:YAG laser에 의한 치수의 초기 조직반응과 calcitonin gene-related peptide (CGRP) 함유 신경섬유의 발현을 비교 평가하고자 시행하였다. Er:YAG laser와 conventional bur를 이용하여 상악 제1대구치의 교합면에 와동을 형성하였다. 와동형성 후 48시간 후에 조직을 처리하여 hematoxylin-eosin 염색과 CGRP 면역조직화학을 시행한 후 다음과 같은 결과를 얻었다. 1. Conventional bur를 이용하여 삭제한 경우, 상아질 바닥면은 균열이 없는 평면의 균일한 모양을 보였으며, Er:YAG laser의 경우에는 상아질의 면은 bur에 비해서 파동형의 불규칙적인 형태와 바닥면에서 상아세관의 주행방향을 따라 미세한 균열을 보였다. 2. Er:YAG laser에 의해 손상 받은 치수에서는 혈관확장과 노출된 치수의 손상부위를 따라 bur에서 손상 받은 치수보다 더 많은 염증세포의 침윤과 상아모세포층의 파괴를 보였다. 3. 노출손상 후에 치수신경 말단에서 CGRP 발현은 bur와 laser군 모두에서 손상을 받지 않은 정상 치수에 비해서 증가된 양상을 보였으며, 그 증가된 정도는 laser의 경우에서 더 강하였다. 위의 결과들을 종합하면, Er:YAG laser가 치수손상을 최소화하여 적용되면 치아우식 제거와 와동형성과 같은 여러 수복치료 시 유용하게 사용될 수 있으리라 사료된다.

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