• Title/Summary/Keyword: crRNA

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Study on the Structure of 5S rRNA from Pseudomonas alcaligenes by Metallotripeptides (금속펩타이드를 이용한 Pseudomonas alcaligenes의 5S rRNA의 구조 연구)

  • Kim, Hee-Joung;Kim, Si-Wouk;Koh, Moon-Joo
    • Journal of the Korean Chemical Society
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    • v.46 no.1
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    • pp.46-51
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    • 2002
  • The recognition and cleavage of 5S rRNA from P. alcaligenes by metallopeptides to the form $Ni(II){\cdot}Gly$-Gly-His(Arg)COOH and $Cu(II){\cdot}Gly$-Gly-His(Arg)COOH were investigated. The results of RNA cleavage analyses suggest that metallopeptides selectively target the unpaired or unstably paired bases of stem-loop structure of 5S rRNA. The selectivity of metallopeptides was little affected by the species of metal ion, Ni(II) or Cu(II). When the result of cleavage by metallopeptides was compared with that of by metal complexes M(II)CR, the recognition by metallopeptides was more selective and structure specific. The cleavage data by metallopeptides and other metal complexes were used to probe the secondary structure of 5S rRNA from P. alcaligenes.

Anti-inflammatory effect of chloroform fraction of Coptidis rhizoma on the production of inflammatory mediators from LPS-stimulated BV2 microglial cells (황련 클로로포름 분획물의 뇌신경소교세포로부터 염증매개물질 생성억제 효능 연구)

  • Park, Yong-Ki;Lee, Kyuong-Yeol
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.109-116
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    • 2007
  • Objectives : In the present study, we investigated anti-inflammatory effects of chloroform fraction of Coptidis rhizoma (CR-C) on the production of inflammatory mediators such as nitric oxide (NO) and proinflammatory cytokines, tumor necrosis factor-alpha (TNF-${\alpha}$) and interleukin-1beta (IL-1${\beta}$) in LPS-stimulated BV2 microglial cells. Methods : Copriditis rhizoma was extracted with 80% methanol, and then extracted with chloroform. BV2 cells were pre-treated with CR-C, and stimulated with LPS. The cytotoxicity was determined by MTT assay. The production of NO and cytokines was measured by Griess assay and ELISA. The mRNA expression of inducible nirtic oxide synthase (iNOS) and cytokines were determined by RT-PCR. Results : CR-C significantly inhibited the production of NO. TNF-${\alpha}$ and IL-1${\beta}$ in a dose-dependent manner in LPS-stimulated BV2 cells. In addition, CR-C suppressed the mRNA expressions of iNOS and inflammatory cytokines induced by LPS stimulation. These results indicate that CR-C was involved in anti-inflammatory effects in activated microglia. Conclusion : The present study suggests that chloroform extract of Coptidis rhizoma can be useful as a potential anti-inflammatory agent for treatment of various neurodegenerative diseases.

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Effect of Chromium Picolinate on Growth Performance, Carcass Characteristics, Serum Metabolites and Metabolism of Lipid in Pigs

  • Xi, Gang;Xu, Zirong;Wu, Si-hung;Chen, Shijiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.258-262
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    • 2001
  • The study was conducted to evaluate the effects of chromium picolinate (CrP) on growth, carcass characteristics and serum metabolites in growing-finishing pigs. A total of 96 Landrace$\times$Yorkshire$\times$Duroc hybrid pigs, initial live weight about $38.12{\pm}00kg$, were randomly assigned to 2 groups (16 pigs per pen, 3 pens per group), each group had 48 pigs with an equal number of barrows and gilts. The pigs were fed the diet with or without $200{\mu}g/kg$ Cr from CrP. The results indicated that the addition of $200{\mu}g/kg$ CrP increased ADG by 3.58% and decreased feed conversion rate (FCR) by 3.00% compared to the control group. Pigs fed CrP had 7.58% (p<0.05) higher carcass lean percentage, 15.55% (p<0.05) larger longissimus muscle area (LMA) and 10.90% (p<0.05) lower back fat thickness, 15.17% (p<0.05) lower carcass fat percentage. In addition, the IGF-I level in serum was elevated by 79.20% (p<0.05), the Insulin and cortisol level decreased by 27.35% (p<0.05) and 34.58% (p<0.05) respectively with supplementation of CrP. Analysis of subcutaneous fat (10th rib) showed that the activity of hormone sensitive lipase (HSL) increased by 79.58% (p<0.05) and the activities of isocitrate dehydrogenase (ISD) and malate dehydrogenase (MDH) decreased significantly by 15.06% (p<0.05) and 54.53% (p<0.05) respectively in the $200{\mu}g/kg$ CrP group. The concentration of RNA, RNA/DNA in LMA increased by 31.89% (p<0.05) and 5.41% (p<0.05) respectively with the addition of CrP. These results suggest that CrP reduced fat deposits by decreasing lipogenic enzyme activities and increasing HSL activity and may have promoted muscle anabolic metabolism through elevated IGF-I levels.

Effect of Coptidis Rhizoma Steamed with Rice Wine on Gastroduodenal Mucosa of Mouse through Inhibiting iNOS Activation (주증황련(酒蒸黃連)이 iNOS 활성 억제를 통해 생쥐 위.십이지장 점막에 미치는 영향)

  • Kim, Myung-Ho;Lim, Seong-Woo
    • The Journal of Internal Korean Medicine
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    • v.35 no.3
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    • pp.262-273
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    • 2014
  • Objectives: This study was carried out to investigate the protective effect of Coptidis Rhizoma steamed with rice wine (CR) against gastroduodenal mucosal injury through inhibiting inducible nitric oxide synthase (iNOS) activation. Methods: In in vitro experiment, LPS-induced RAW 264.7 macrophages were treated with CR(0.4, 0.6, 0.8, 1.0 mg/ml) and iNOS mRNA expression and nitric oxide (NO) production were measured. In in vivo experiment normal group mice were treated with neither ethanol nor CR. Both control and sample group mice were orally administrated with ethanol. Five hours after ethanol administration control group mice were orally administrated with distilled water, sample group mice were orally administrated with CR. After three days administration, gastroduodenal mucosa of mice was observed histopathologically and iNOS, nuclear factor-kappa B (NF-${\kappa}B$) activation were observed immunohistochemically. Results: In in vitro experiment iNOS mRNA expression and NO production in LPS-induced RAW 264.7 macrophages were decreased by CR dose-dependently. In in vivo experiment, gastroduodenal mucosal injury was repaired by CR and iNOS, NF-${\kappa}B$ activation in gastroduodenal mucosa were decreased by CR. Conclusions: Coptidis Rhizoma steamed with rice wine has a protective effect against gastroduodenal mucosal injury through inhibiting iNOS activation.

생쥐 초기 배아에서 Aquaporin 8과 9의 발현에 관한 연구

  • 신현상;계명찬;강수만;이성은;이지원;강한승;김문규
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.68-68
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    • 2003
  • Aquaporin은 막관통 통로 단백질(transmembrane channel protein)로서, 삼투압의 농도구배에 따라 세포막을 가로질러 물분자를 이동시키는 기능을 하고 있다. 포유류 초기배아에서 포배강 형성은 영양외배엽세포에서 $Na^+ / K^+$ATPase에 의한 이온 농도 구배가 형성되면 auqaporin에 의해 물이 포배강으로 유입되면서 이루어진다. 본 연구에서는 생쥐 초기배아에서 반정량적인 역전사 중합효소 연쇄반응 방법(semi-quantitative RT-PCR)과 실시간 역전사 중합효소 연쇄반응 방법(real-time RT-PCR)을 통하여 AQP8과 9의 mRNA발현을 조사하고 다중 면역형광현미경 방법(confocal immunofluorescence microscopy)을 통해 단백질 발현양상을 분석하였다. AQP8 mRNA는 상실기까지 발현되지 않다가 포배기에 이르러 발현되었고 AQP9 mRNA는 수정란에서부터 발현되어 포배기에는 유의할 정도로 증가하였다. 따라서 AQP8 mRNA는 배아유전자가 활성화되어 나타나는 것이고 AQP9 mRNA는 모계유전자 기원임을 알 수 있었다. AQP8 단백질은 상실배 단계까지 발현되지 않다가 포배시기에 영양외배엽세포사이의 접합면에 발현되었고 AQP9 단백질은 상실배 시기에 할구 사이의 인접 부위에서 강하게 발현되었다가 포배시기에는 세포간의 접합면에 약하게 발현하는 경향을 나타내었다. 실시간 역전사 중합효소 연쇄반응 방법으로 조사한 결과 포배에서 물과 글리세롤을 통과시키는 AQP9는 mRNA의 발현양이 AQP8보다 약 4배 정도 많았다. 또한 포배기에 이르러서야 물만을 통과시키는 AQP8의 발현이 나타나는 것을 보아 포배강 형성시 외부에서 영양외배엽을 통해 포배강으로 유입되는 물의 이동(trans- trophectodermal water movements)에 AQP9보다 AQP8이 더 중요하게 관여할 것으로 사료된다., K, Pb, Cd, Cr, Co, Cu, Ni)을 측정하였다. 실험 조건1의 결과로서 각 국의 유아용 일회용 기저귀의 중금속 함량은 거의 유사한 경향을 나타내었으며 Cr, Zn, Pb, Ni, Mn, Mg, Li, K는 detection limit(2 ppm) 이하였고, Cd, Fe, Co, Cu, Ca, Al, Sr는 검출되었지만 기준치 이하였다. 실험 조건2의 결과로서 측정 항목(Cr, Sb, Cd, Pb, Ni, Co, Cu)중 Cr, Cd, Ni, Cu는 detection limit(0.1 ppm) 이하였고, Sb, Pb, Co는 검출되었지만 기준치 이하였다.았다. 4%의 경우에는 8$0^{\circ}C$이하로 온도를 낮추는 것이 좋은 상태를 나타내었다. 이와 같은 결과는 일반적으로 화학적 레팅을 4%, 7%에서한 선행결과와 상당히 다른 결과이다.염 농도가 증가할수록 감소 현상을 보였다.X>, 75BG30은 8.6$\mu\textrm{m}$, 75BG40은 7.02$\mu\textrm{m}$로 나타났다. 따라서 경화제 양에 관계없이 10$\mu\textrm{m}$ 이하로 나타나, 경화제 10$m\ell$만으로 미세한 크기를 얻을 수 있음을 알 수 있다. 젤리 강도 변화에 따른 차이는 300BF는 78.09$\mu\textrm{m}$ 300BG는 56.32$\mu\textrm{m}$로, 75BF나 75BG에 비하여 현저히 증가하여, 젤라틴의 젤리 강도는 캡슐 제조 조건의 주요한 변수임을 알 수 있다.추출물 투여시 혈당강하 및 혈중콜레스테롤 강하가 나타났으며, 상엽복합추출물 투여와 운동을 병행시 이러한 감소 효과가 더 뚜렷하게 나타났다.교육의 적임자로 보는 시각이 비교적 높았고 약 1/2정도는 영양교육에 참여하겠다는 의지를 가지고 있을 뿐만 아니라 실제로 영양지도를

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Correlation of Gene Expression between Adiponectin and Glucose Transporter 4 in Mouse Adipose Tissue (생쥐 지방조직에서의 아디포넥틴과 포도당수송체-4 유전자 발현의 상관관계)

  • Lee, Yong-Ho
    • Journal of Life Science
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    • v.24 no.8
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    • pp.895-902
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    • 2014
  • Adiponectin has been known to improve insulin sensitivity and elicit glucose uptake via increased glucose transporter 4 (GLUT4) translocation. In the current study, mRNA expression levels of adiponectin and GLUT4 were measured in subcutaneous adipose tissue from C57BL/6 mice fed normal (ND) or high-fat diet (HFD) until 16, 26, 36, 47, or 77 weeks of age starting from 6 weeks of age. Expression levels were also measured in mice with calorie restriction (CR) and in thiazolidinedione (TZD) treated mice. Using quantitative real-time PCR, we demonstrated that GLUT4 expression in adipose tissue significantly decreased in HFD mice groups and increased in CR (p<0.05) and TZD (p=0.007) groups while there was no difference in adiponectin mRNA expression levels between experimental and control groups. General linear regression models were used to assess the association of gene expression levels between adiponectin and GLUT4 and to determine whether adiponectin affects GLUT4 transcription. mRNA expression levels of adiponectin and GLUT4 are significantly associated each other in mice fed a ND (p<0.0001) or HFD (p<0.0001), in groups separated into each age and diet, and CR group (p=0.002), but not in TZD group (p=0.73). These results demonstrated that gene expression of adiponectin and GLUT4 is strongly associated, suggesting that there is a common regulatory mechanism for adiponectin and GLUT4 gene expression and/or adiponectin has a direct role in GLUT4 gene expression in adipose tissue.

Direct and Indirect Reduction of Cr(VI) by Fermentative Fe(III)-Reducing Cellulomonas sp. Strain Cellu-2a

  • Khanal, Anamika;Hur, Hor-Gil;Fredrickson, James K.;Lee, Ji-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1519-1525
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    • 2021
  • Hexavalent chromium (Cr(VI)) is recognized to be carcinogenic and toxic and registered as a contaminant in many drinking water regulations. It occurs naturally and is also produced by industrial processes. The reduction of Cr(VI) to Cr(III) has been a central topic for chromium remediation since Cr(III) is less toxic and less mobile. In this study, fermentative Fe(III)-reducing bacterial strains (Cellu-2a, Cellu-5a, and Cellu-5b) were isolated from a groundwater sample and were phylogenetically related to species of Cellulomonas by 16S rRNA gene analysis. One selected strain, Cellu-2a showed its capacity of reduction of both soluble iron (ferric citrate) and solid iron (hydrous ferric oxide, HFO), as well as aqueous Cr(VI). The strain Cellu-2a was able to reduce 15 μM Cr(VI) directly with glucose or sucrose as a sole carbon source under the anaerobic condition and indirectly with one of the substrates and HFO in the same incubations. The heterogeneous reduction of Cr(VI) by the surface-associated reduced iron from HFO by Cellu-2a likely assisted the Cr(VI) reduction. Fermentative features such as large-scale cell growth may impose advantages on the application of bacterial Cr(VI) reduction over anaerobic respiratory reduction.

Hexavalent Chromium Reduction by Bacteria from Tannery Effluent

  • Batool, Rida;Yrjala, Kim;Hasnain, Shahida
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.547-554
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    • 2012
  • Chromium is generated from several industrial processes. It occurs in different oxidation states, but Cr(III) and Cr(VI) are the most common ones. Cr(VI) is a toxic, soluble environmental contaminant. Some bacteria are able to reduce hexavalent chromium to the insoluble and less toxic Cr(III), and thus chromate bioremediation is of considerable interest. An indigenous chromium-reducing bacterial strain, Rb-2, isolated from a tannery water sample, was identified as Ochrobactrum intermedium, on the basis of 16S rRNA gene sequencing. The influence of factors like temperature of incubation, initial concentration of Cr, mobility of bacteria, and different carbon sources were studied to test the ability of the bacterium to reduce Cr(VI) under variable environmental conditions. The ability of the bacterial strain to reduce hexavalent chromium in artificial and industrial sewage water was evaluated. It was observed that the mechanism of resistance to metal was not due to the change in the permeability barrier of the cell membrane, and the enzyme activity was found to be inductive. Intracellular reduction of Cr(VI) was proven by reductase assay using cell-free extract. Scanning electron microscopy revealed chromium precipitates on bacterial cell surfaces, and transmission electron microscopy showed the outer as well as inner distribution of Cr(VI). This bacterial strain can be useful for Cr(VI) detoxification under a wide range of environmental conditions.

Effects of Dietary Chromium Methionine on Growth Performance, Carcass Composition, Meat Colour and Expression of the Colour-related Gene Myoglobin of Growing-finishing Pigs

  • Li, Y.S.;Zhu, N.H.;Niu, P.P.;Shi, F.X.;Hughes, C.L.;Tian, G.X.;Huang, R.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.7
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    • pp.1021-1029
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    • 2013
  • To investigate the effect of dietary chromium (Cr) as Cr methionine (CrMet) on growth performance, carcass traits, pork quality, meat colour and expression of meat colour-related genes in growing-finishing pigs, 189 crossbred Duroc${\times}$(Landrace${\times}$Yorkshire) growing-finishing pigs (male, castrated, average initial BW $74.58{\pm}1.52$ kg) were selected and randomly allocated into four groups. Dietary treatments per kg of feed were as follows: 0 (CT), 0.3 mg/kg (T1), 0.6 mg/kg (T2) and 0.9 mg/kg (T3) Cr (in the form of CrMet; as-fed basis), and each treatment was replicated five times with 8 to 10 pigs per replicate pen. During the 28 d of the experiment, both the ADG and the ADFI increased linearly (p<0.05) as the level of dietary Cr increased. The F/G ratio decreased linearly (p<0.05). As dietary Cr increased, loin muscle areas (linear, p = 0.013) and average backfat thickness (linear, p = 0.072) decreased. Shear force (linear, p = 0.070) and Commission Internationale de I'$\acute{E}$clairage (CIE) redness (quadratic, p = 0.028) were increased. In addition, CIE Lightness (quadratic, p = 0.053) were decreased as dietary Cr increased. As dietary Cr increased, total myglobin (Mb) content (quadratic, p = 0.015) and the mb mRNA levels (quadratic, p = 0.046) in longissimus muscles of pigs were up-regulated. In conclusion, supplementation of dietary Cr improved growth and meat colour, but increased shear force and decreased IMF reduced palatability of longissimus muscles. Moreover, the increasing total Mb content and mb mRNA levels indicated that CrMet dietary supplementation may improve meat colour via up-regulating expression of the mb gene.

Harnessing CRISPR-Cas adaptation for RNA recording and beyond

  • Gyeong-Seok Oh;Seongjin An;Sungchul Kim
    • BMB Reports
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    • v.57 no.1
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    • pp.40-49
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    • 2024
  • Prokaryotes encode clustered regularly interspaced short palindromic repeat (CRISPR) arrays and CRISPR-associated (Cas) genes as an adaptive immune machinery. CRISPR-Cas systems effectively protect hosts from the invasion of foreign enemies, such as bacteriophages and plasmids. During a process called 'adaptation', non-self-nucleic acid fragments are acquired as spacers between repeats in the host CRISPR array, to establish immunological memory. The highly conserved Cas1-Cas2 complexes function as molecular recorders to integrate spacers in a time course manner, which can subsequently be expressed as crRNAs complexed with Cas effector proteins for the RNA-guided interference pathways. In some of the RNA-targeting type III systems, Cas1 proteins are fused with reverse transcriptase (RT), indicating that RT-Cas1-Cas2 complexes can acquire RNA transcripts for spacer acquisition. In this review, we summarize current studies that focus on the molecular structure and function of the RT-fused Cas1-Cas2 integrase, and its potential applications as a directional RNA-recording tool in cells. Furthermore, we highlight outstanding questions for RT-Cas1-Cas2 studies and future directions for RNA-recording CRISPR technologies.