• Title/Summary/Keyword: 6S RNA

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Effects of Chaenomelis Fructus Extract on the Alzheimer's Disease Mice Model Induced by $\betaA$ (목과의 $\betaA$로 유도된 Alzheimer's Disease 생쥐 모델에 미치는 영향)

  • Jung In Chul;Lee Sang Ryong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1795-1804
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    • 2004
  • This research investigated the effect of the Chaenomelis fructus(CMF) on Alzheimer's disease. The effects of the CMF extract on the behavior in the Morris water maze experiment; the expression of IL-1β, TNF-α, ROS on the microglial cell; IL-1β mRNA, TNF-α mRNA, CD68/GFAP and MDA on the brain tissue; the infarction area of the hippocampus, and brain tissue injury in the mice with Alzheimer's disease induced by βA were investigated. The CMF extract group showed a significant inhibitory effect on the memory deficit on the mice with Alzheimer's disease induced by βA in the Morris water maze experiment. The CMF extract group suppressed the over-expression of IL-1β, TNF-α, IL-1β and TNF-α mRNA, ROS, MDA, CD68/GFAP in the mice with Alzheimer's disease induced by βA. The CMF extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by [3A. This study suggest that CMF may be effective for the prevention and treatment of Alzheimer's disease.

Regulation of S100G Expression in the Uterine Endometrium during Early Pregnancy in Pigs

  • Choi, Yo-Han;Seo, Hee-Won;Shim, Jang-Soo;Kim, Min-Goo;Ka, Hak-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.44-51
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    • 2012
  • Calcium ions play an important role in the establishment and maintenance of pregnancy, but molecular and cellular regulatory mechanisms of calcium ion action in the uterine endometrium are not fully understood in pigs. Previously, we have shown that calcium regulatory molecules, transient receptor potential vanilloid type 5 (TRPV6) and calbindin-D9k (S100G), are expressed in the uterine endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner, and that estrogen of conceptus origin increases endometrial TRPV6 expression. However, regulation of S100G expression in the uterine endometrium and conceptus expression of S100G has been not determined during early pregnancy. Thus, we investigated regulation of S100G expression by estrogen and interleukin-$1{\beta}$ (IL1B) in the uterine endometrium and conceptus expression of S100G during early pregnancy in pigs. We obtained uterine endometrial tissues from day (D) 12 of the estrous cycle and treated with combinations of steroid hormones, estradiol-$17{\beta}$ ($E_2$) and progesterone ($P_4$), and increasing doses of IL1B. Real-time RT-PCR analysis showed that $E_2$ and IL1B increased S100G mRNA levels in the uterine endometrium, and conceptuses expressed S100G mRNA during early pregnancy, as determined by RT-PCR analysis. To determine if endometrial expression of S100G mRNA during the implantation period was affected by the somatic cell nuclear transfer (SCNT) procedure, we compared S100G mRNA levels in the uterine endometrium from gilts with SCNT-derived conceptuses with those from gilts with conceptuses derived from natural mating on D12 of pregnancy. Real-time RT-PCR analysis showed that levels of S100G mRNA in the uterine endometrium from gilts carrying SCNT-derived conceptuses was significantly lower than those from gilts carrying conceptuses derived from natural mating. These results showed that S100G expression in the uterine endometrium was regulated by estrogen and IL1B of conceptus origin, and affected by the SCNT procedure during early pregnancy. These suggest that conceptus signals regulate S100G, an intracellular calcium transport protein, for the establishment of pregnancy in pigs.

Cold Shock Response and Low Temperature Stable Transcript of DEAD-box RNA Helicase in Bacillus subtilis (DEAD-box RNA Helicase 유전자가 결핍된 Bacillus subtilis의 저온 충격 반응성과 저온 안정성 전사물)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.289-294
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    • 2011
  • We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

Occurrence of Indicator Bacteria and Identification of Total Coliforms Using 16S rRNA Gene in Drinking Spring Water in Seoul (서울 약수터의 지표세균 분포 및 16S rRNA 염기서열을 이용한 총대장균군 동정 및 계통분석)

  • Yoon, Tae-Ho;Lee, Hyang;Choi, Geum-Sook;Lee, Seung-Joo;Lee, Mok-Young;Eo, Soo-Mi
    • Journal of Environmental Health Sciences
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    • v.39 no.6
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    • pp.513-521
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    • 2013
  • Objectives: This study was performed in order to detect indicator bacteria in drinking spring water (DSW) samples in Seoul Metropolitan City, and to identify their genus through 16S rRNA sequencing and then assessing the genetic relation of their strains. Methods: For indicator bacteria detection and identification of total coliforms, we analyzed DSW between the spring and summer seasons. In particular, DSW samples were chosen from sites repeatedly found unsatisfactory in recent years. Results: Heterotrophic plate counts of DSW in the spring and summer season were investigated in the range of 0-550 and 0-800 CFU/mL, respectively. Total coliforms of these were 0-1,900 and 0-2,100 CFU/100mL, fecal coliforms were 0-600 and 0-550 CFU/100mL, and Escherichia coli were 0-7 and 0-326 MPN/100mL. The detection ratio of fecal pollution indicators and that of fecal coliforms increased to 58.6% in the summer from 12.5% in the spring and Escherichia coli increased to 51.4% from 4.7%. As a result of genetic analysis on the isolated bacteria, the genus of total coliforms was classified in the order of Enterobacter spp. 12.7%, Serratia spp. 7.3%, E. hermanii 6.4%, Rahnella spp. 5.5%, Hafnia spp. 4.5%, Escherichia coli 3.6%, Klebsiella spp. 3.6% in the spring season. In the summer season, it was classified in order of Klebsiella spp. 16.6%, Enterobacter spp. 13.0%, Escherichia coli 11.0%, Serratia spp. 8.6%, Raoultella spp. 7.0%, Kluyvera spp. 5.6% and Citrobacter spp. 3.0%. Conclusions: The increase of fecal pollution in summer indicates that special attention to drinking DSW is required.

Antibacterial Activity against Salmonella enteritidis JK-15 and LPS Changes Caused by Rose Flower Extracts (장미꽃 추출물에 의한 식중독 세균 Salmonella enteritidis JK-15에 대한 살균활성 및 그에 따른 LPS 변화)

  • Song, You-Jin;Cho, Yun-Seok;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.318-323
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    • 2009
  • The aim of this work was to investigate the antibacterial effect of the food-poisoning bacterium, Salmonella enteritidis JK-15 exposed to rose extracts. Initially, the isolate S. enteritidis JK-15 was enriched and isolated from stale food. BIOLOG and 16S rRNA analyses revealed that strain S. enteritidis JK-15 was 98% similar to the S. enteritidis species cluster; therefore we have designated this strain as S. enteritidis JK-15. Bactericidal effects of S. enteritidis JK-15 exposed to rose extracts ranging from 5 mg/ml to 100 mg/ml were monitored, and complete bactericidal effects were achieved within 6 h at 100 mg/ml and 12 h at 50 mg/ml, respectively. SDSPAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain S. enteritidis JK-15 treated to different concentrations and exposing periods of rose extracts in exponentially growing cultures. Scanning electron microscopic analysis, demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with rose extracts.

Antibiofilm Activity and Binding Specificity of Polyclonal DNA Aptamers on Staphylococcus aureus and Escherichia coli

  • Arizah Kusumawati;Apon Zaenal Mustopa;Rifqiyah Nur Umami;Adi Santoso;I Wayan Teguh Wibawan;Agus Setiyono;Mirnawati Bachrum Sudarwanto
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.328-336
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    • 2022
  • Aptamers are short, chemically synthesized, single-stranded DNA or RNA oligonucleotides that fold into unique three-dimensional structures. In this study, we aim to determine the antibiofilm activity and binding specificity of the six polyclonal DNA aptamers (S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20) on Staphylococcus aureus BPA-12 and Escherichia coli EPEC 4. Aptamer S15K6 showed the highest percentage of antibiofilm activity against S. aureus BPA-12 (37.4%) as shown by the lowest OD570 value of 0.313. Aptamer S15K20 showed the highest percentage of antibiofilm activity against E. coli EPEC 4 (15.4%) as shown by the lowest OD570 value of 0.515. Aptamers S15K13 and S15K20 showed antibiofilm activities against both S. aureus BPA-12 and E. coli EPEC4, and thus potentially have broad reactivity. Furthermore, based on the binding capacity and Kd values from our previous study, the binding specificity assay of selected polyclonal DNA aptamers (S15K3 and S15K15) against S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, S. agalactiae, E. coli MHA-6, and Listeria monocytogenes were performed using qPCR. Aptamers S15K3 and S15K15 showed specific binding to S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, and S. agalactiae, but could not bind to E. coli MHA-6 and L. monocytogenes. Therefore, this study showed that the polyclonal DNA aptamers have antibiofilm activity and were able to bind to S. aureus BPA-12 and E. coli EPEC 4 bacteria.

Construction of a Hammerhead Ribozyme that Cleaves Rice Black-Streaked Dwarf Virus RNA (흑조위축병 바이러스 RNA를 절단하는 망치머리형 라이보자임의 제작)

  • Kim, Ju-Kon;Sohn, Seong-Han;Lee, Sug-Soon;Hwang, Young-Soo;Park, Jong-Sug
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.522-527
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    • 1995
  • To develop an antiviral agent for the rice black-streaked dwarf virus (RBSDV), a hammerhead type ribozyme, which has a potential target site on the genome segment 3, was designed. Oligonucleotides for the ribozyme and its substrate were synthesized, annealed, and cloned into a plasmid pBluescript II KS(+). Ribozyme and substrate RNAs were then synthesized by in vitro transcription with $T_3$ RNA polymerase, obtaining RNAs in expected size, 193 and 182 nucleotides, respectively. The substrate RNA was efficiently cleaved into two fragments when incubated with the ribozyme at $55^{\circ}C$, while the cleavage was not detected at $37^{\circ}C$. In addition, the segment 3 RNA of RBSDV was also cleaved into two fragments by the same ribozyme at $55^{\circ}C$. Taken together, our results demonstrated that the hammerhead ribozyme has an in vitro endonucleolytic activity and may be used as an antiviral agent in transgenic plants.

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Anaerobic Ammonium-Oxidizing Bacteria in Cow Manure Composting

  • Wang, Tingting;Cheng, Lijun;Zhang, Wenhao;Xu, Xiuhong;Meng, Qingxin;Sun, Xuewei;Liu, Huajing;Li, Hongtao;Sun, Yu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1288-1299
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    • 2017
  • Composting is widely used to transform waste into valuable agricultural organic fertilizer. Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the global nitrogen cycle, but their role in composting remains poorly understood. In the present study, the community structure, diversity, and abundance of anammox bacteria were analyzed using cloning and sequencing methods by targeting the 16S rRNA gene and the hydrazine oxidase gene (hzo) in samples isolated from compost produced from cow manure and rice straw. A total of 25 operational taxonomic units were classified based on 16S rRNA gene clone libraries, and 14 operational taxonomic units were classified based on hzo gene clone libraries. The phylogenetic tree analysis of the 16S rRNA gene and deduced HZO protein sequences from the corresponding encoding genes indicated that the majority of the obtained clones were related to the known anammox bacteria Candidatus "Brocadia," Candidatus "Kuenenia," and Candidatus "Scalindua." The abundances of anammox bacteria were determined by quantitative PCR, and between $2.13{\times}10^5$ and $1.15{\times}10^6$ 16S rRNA gene copies per gram of compost were found. This study provides the first demonstration of the existence of anammox bacteria with limited diversity in cow manure composting.

Molecular Characterization and Expression Analysis of Interferon Regulatory Factor 8 (IRF8) in the Black Rockfish Sebastes schlegelii (조피볼락(Sebastes schlegelii) Interferon Regulatory Factor 8 (IRF8)의 분자유전학적 특성 및 발현 분석)

  • Yang, Hyerim;Kwon, Hyukjae;Lee, Seongdo;Bathige, S.D.N.K;Kim, Myoung-Jin;Lee, Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.3
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    • pp.302-310
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    • 2017
  • Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mRNA expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cDNA sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kDa and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mRNA was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mRNA expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qRT-PCR results showed that SsIRF8 mRNA expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mRNA in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents.

Changes of RNA and Protein During Callus Induction and Plant Regeneration from Perilla frutescens (들깨로부터 Callus의 유기와 재분화에 따른 단백질 및 RNA의 변화)

  • 정상훈;양선경;김현경;정대수;조영수;김도훈
    • Journal of Life Science
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    • v.9 no.1
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    • pp.29-34
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    • 1999
  • Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

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