• Title/Summary/Keyword: De novo

Search Result 421, Processing Time 0.034 seconds

Dynamic DNA Methylation Change of Dnmt1o 5'-Terminal Region during Preimplantation Development of Cloned Pig (돼지 체세포 복제란 초기발달 과정 중 Dnmt1o 상류 영역의 다이내믹한 DNA 메틸화 변화)

  • Ko, Yeoung-Gyu;Kim, Sung-Woo;Cho, Sang-Rae;Do, Yoon-Jung;Kim, Jae-Hwan;Kim, Sang-Woo;Kim, Hyun;Park, Jae-Hong;Park, Soo-Bong
    • Reproductive and Developmental Biology
    • /
    • v.36 no.1
    • /
    • pp.7-12
    • /
    • 2012
  • DNA methyltransferase 1 (Dnmt1) gene contains three different isoform transcripts, Dnmt1s, Dnmt1o, and Dnmt1p, are produced by alternative usage of multiple first exons. Dnmt1o is specific to oocytes and preimplantation embryos, whereas Dnmt1s is expressed in somatic cells. Here we determined that porcine Dnmt1o gene had differentially methylated regions (DMRs) in 5'-flanking region, while those were not found in the Dnmt1s promoter region. The methylation patterns of the porcine Dnmt1o/Dnmt1s DMRs were investigated using bisulfite sequencing and pyrosequencing analysis through all preimplantation stages from one cell to blastocyst stage in in vivo or somatic cell nuclear transfer (SCNT). The Dnmt1o DMRs contained 8 CpG sites, which located in -640 bp to -30 bp upstream region from transcription start site of the Dnmt1o gene. The methylation status of 5 CpGs within the Dnmt1o DMRs were distinctively different at each stage from one-cell to blastocyst stage in the $in$ $vivo$ or SCNT, respectively. 55.62% methylation degree of the Dnmt1o DMRs in the $in$ $vivo$ was increased up to 84.38% in the SCNT embryo, moreover, $de$ $novo$ methylation and demethylation occurred during development of porcine embryos from the one-cell stage to the blastocyst stage. However, the DNA methylation states at CpG sites in the Dnmt1s promoter regions were hypomethylated, and dramatically not changed through one-cell to blastocyst stage in the $in$ $vivo$ or SCNT embryos. In the present study, we demonstrated that the DMRs in the promoter region of the porcine Dnmt1o was well conserved, contributing to establishment and maintenance of genome-wide patterns of DNA methylation in early embryonic development.

Characterization and Functional Study of PyrR Orthologues from Genome Sequences of Bacteria (세균 게놈 유래성 PyrR Orthologue의 기능 분석)

  • 김사열;조현수;설경조;박승환
    • Microbiology and Biotechnology Letters
    • /
    • v.31 no.2
    • /
    • pp.103-110
    • /
    • 2003
  • The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.

A Case of Interstitial Pneumonitis Caused by Leflunomide (Leflunomide에 의한 약물유발 간질성 폐렴 1예)

  • Lee, Jung Hwa;Cheon, Won Seok;Seo, Young Il;Eom, Kwang-Seok;Jang, Seung Hun;Bahn, Joon-Woo;Kim, Dong-Gyu;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
    • /
    • v.58 no.1
    • /
    • pp.83-88
    • /
    • 2005
  • Leflunomide is a new disease modifying anti rheumatic drug (DMARD) for the treatment of active rheumatoid arthritis. Its mechanism of action differs from other DMARDs in that it inhibits the de novo pyrimidine synthesis by inhibiting dihydroorotate dehydrogenase and therefore prevents the proliferation of activated lymphocytes. As it has been prescribed worldwide, there is a great deal of much concerns regarding its potential adverse effects. Because leflunomide has an active metabolite with a long elimination half-life of approximately 2 weeks, serious adverse reactions may occur even after the leflunomide treatment has been stopped. The profile of serious reactions includes liver dysfunction, hematological disorders, severe skin reactions and respiratory dysfunction. Respiratory dysfunctions with leflunomide therapy are very rare and its incidence is lower than that of methotrexate therapy. However, there are reports in Japan showing that 5 patients died of interstitial pneumonitis and another 11 patients developed serious lung complications associated with leflunomide. This suggests the possibility of fatal respiratory toxicity of leflunomide. There are no reports of interstitial pneumonitis associated with leflunomide in Korea. We report a case of a 62-year old woman who developed interstitial pneumonitis, which might have been induced by leflunomide during the treatment of rheumatoid arthritis.

Transcriptome profiling and comparative analysis of Panax ginseng adventitious roots

  • Jayakodi, Murukarthick;Lee, Sang-Choon;Park, Hyun-Seung;Jang, Woojong;Lee, Yun Sun;Choi, Beom-Soon;Nah, Gyoung Ju;Kim, Do-Soon;Natesan, Senthil;Sun, Chao;Yang, Tae-Jin
    • Journal of Ginseng Research
    • /
    • v.38 no.4
    • /
    • pp.278-288
    • /
    • 2014
  • Background: Panax ginseng Meyer is a traditional medicinal plant famous for its strong therapeutic effects and serves as an important herbal medicine. To understand and manipulate genes involved in secondary metabolic pathways including ginsenosides, transcriptome profiling of P. ginseng is essential. Methods: RNA-seq analysis of adventitious roots of two P. ginseng cultivars, Chunpoong (CP) and Cheongsun (CS), was performed using the Illumina HiSeq platform. After transcripts were assembled, expression profiling was performed. Results: Assemblies were generated from ~85 million and ~77 million high-quality reads from CP and CS cultivars, respectively. A total of 35,527 and 27,716 transcripts were obtained from the CP and CS assemblies, respectively. Annotation of the transcriptomes showed that approximately 90% of the transcripts had significant matches in public databases.We identified several candidate genes involved in ginsenoside biosynthesis. In addition, a large number of transcripts (17%) with different gene ontology designations were uniquely detected in adventitious roots compared to normal ginseng roots. Conclusion: This study will provide a comprehensive insight into the transcriptome of ginseng adventitious roots, and a way for successful transcriptome analysis and profiling of resource plants with less genomic information. The transcriptome profiling data generated in this study are available in our newly created adventitious root transcriptome database (http://im-crop.snu.ac.kr/transdb/index.php) for public use.

Clinical Application of M-number for Aortic Cannulas During Cardiopulmonary Bypass (심폐바이패스시 대동맥캐늘라에 대한 M-NUMBER의 임상 적용)

  • 김원곤;박성식
    • Journal of Chest Surgery
    • /
    • v.29 no.5
    • /
    • pp.510-516
    • /
    • 1996
  • Cardiopulmonary bypass cannulas are usually characterized by the French number. However this de- scription provides only the external diameter of the cannula, which gives no information about the press- ure-flow characteristics of the cannula itself. A standardized system to describe the pressure-flow characteristics of a given cannula has recently been proposed and has been termed the M-number It is reported that the pressure-flow characteristics of a particular cannula can be determined from a novo- gram or chart, if the experimentally derived M-number of the cannula is known. In this regard, we conducted an investigation to analyze correlation between experimentally and clinical y derived M-numbers using three different sizes of pediatric aortic cannulas in fifty cardiac patients on cardiopulmonary bypass. The clinical and experimental M-numbers showed a strong correlation. The clinical M-numbers were typically 0.)5 to 0.55 greater than the experimental M-numbers. The clinical M-numbers also showed an inverse relationship to the temperature change of the patient, most probably due to an increase in blood viscosity from hypothermia. This inverse clinical M-numbersltemperature re- lationship was more marked in higher M-number cannulas. The clinical data obtained in this study suggest that the experimentally derived M-numbers correlated strongly with the clinical performance of the cannula with the significant influence of the temperature.

  • PDF

In vivo protein expression changes in mouse livers treated with dialyzed coffee extract as determined by IP-HPLC

  • Yoon, Cheol Soo;Kim, Min Keun;Kim, Yeon Sook;Lee, Suk Keun
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.40
    • /
    • pp.44.1-44.17
    • /
    • 2018
  • Background: Coffee extract has been investigated by many authors, and many minor components of coffee are known, such as polyphenols, diterpenes (kahweol and cafestol), melanoidins, and trigonelline, to have anti-inflammatory, anti-oxidant, anti-angiogenic, anticancer, chemoprotective, and hepatoprotective effects. Therefore, it is necessary to know its pharmacological effect on hepatocytes which show the most active cellular regeneration in body. Methods: In order to determine whether coffee extract has a beneficial effect on the liver, 20 C57BL/6J mice were intraperitoneally injected once with dialyzed coffee extract (DCE)-2.5 (equivalent to 2.5 cups of coffee a day in man), DCE-5, or DCE-10, or normal saline (control), and then followed by histological observation and IP-HPLC (immunoprecipitation high performance liquid chromatography) over 24 h. Results: Mice treated with DCE-2.5 or DCE-5 showed markedly hypertrophic hepatocytes with eosinophilic cytoplasms, while those treated with DCE-10 showed slightly hypertrophic hepatocytes, which were well aligned in hepatic cords with increased sinusoidal spaces. DCE induced the upregulations of cellular proliferation, growth factor/RAS signaling, cellular protection, p53-mediated apoptosis, angiogenesis, and antioxidant and protection-related proteins, and the downregulations of NFkB signaling proteins, inflammatory proteins, and oncogenic proteins in mouse livers. These protein expression changes induced by DCE were usually limited to the range ± 10%, suggesting murine hepatocytes were safely reactive to DCE within the threshold of physiological homeostasis. DCE-2.5 and DCE-5 induced relatively mild dose-dependent changes in protein expressions for cellular regeneration and de novo angiogenesis as compared with non-treated controls, whereas DCE-10 induced fluctuations in protein expressions. Conclusion: These observations suggested that DCE-2.5 and DCE-5 were safer and more beneficial to murine hepatocytes than DCE-10. It was also found that murine hepatocytes treated with DCE showed mild p53-mediated apoptosis, followed by cellular proliferation and growth devoid of fibrosis signaling (as determined by IP-HPLC), and subsequently progressed to rapid cellular regeneration and wound healing in the absence of any inflammatory reaction based on histologic observations.

Current research on seed oil biosynthesis (식물 종자지방 합성대사 연구의 최근 동향)

  • Kim, Hyun Uk;Lee, Kyeong-Ryeol;Kim, Eun Ha;Jung, Su-Jin;Roh, Kyung Hee;Kang, Han Chul;Kim, Jong-Bum
    • Journal of Plant Biotechnology
    • /
    • v.41 no.1
    • /
    • pp.10-18
    • /
    • 2014
  • Seed oils (triacylglycerols) of plants are used as a source of essential fatty acids and nutrition for human. In addition, triacylglycerols have been used as industrial raw materials and biofuels. Triacylglycerols are mainly accumulated in seeds by complicated biochemical pathways. Fatty acids are synthesized in the plastids and transported into the endoplasmic reticulum for synthesizing triacylglycerols. It is known for a long time that biosynthesis of triacylglycerols is performed by a de novo synthesis, the Kennedy pathway. However recent studies have revealed that phosphatidylcholine, a major component of cell membrane, plays a central role for triacylglycerols biosynthesis. Phosphatidylcholine is a key regulator determining the relative proportions of unsaturated fatty acids in seeds. It may be a major carrier for the fluxes of fatty acids from the plastid to the endoplasmic reticulum. This finding further suggests that studies of the functions of enzymes involved in the fluxes of fatty acids from phosphatidylcholine to triacylglycerols elucidate the specialized subdomains in the endoplasmic reticulum for triacylglycerols biosynthesis.

Effects of Feeding Organic Diets with Different Fatty Acid Composition Ratio on CLA and Fatty Acid Contents in Raw Milk of Holstein-Friesian Dairy Cows (지방산 조성 비율이 다른 유기농 사료 급여가 홀스타인 원유의 CLA 및 지방산 함량에 미치는 영양)

  • Nam, In-Sik;Lim, Yang-Cheon;Nam, Ki-Taeg
    • Korean Journal of Organic Agriculture
    • /
    • v.25 no.1
    • /
    • pp.219-231
    • /
    • 2017
  • The aim of this study was to determine the effects of feeding of different organic diets (based on fatty acid contents) on CLA and fatty acid concentration of raw milk. Total two hundred and ninety Holstein cows were divided into control, treatment 1 and treatment 2 groups according to the parity and milk yield. Control diet was contained higher C16:00, C18:2 and SFA. Treatment 1 diet was higher in C18:1, C18:2, UFA and treatment 2 diet was higher in MUFA and C18:3 and PUFA. The results indicated that the C16:0 concentration in raw milk was greater in the group of treatment 2 than in control and treatment 1 (p<0.05). The concentration of CLA in treatment 2 was higher compared to treatment 1 and control groups (p<0.05). The ration of n-3/n-6 was higher in treatment 2 group compared with control and treatment 1 (p<0.05). In conclusion, feeding 100% of grass feed for dairy cows increases CLA and n-3 content in milk. From now on, basis on our study, development of functional milk will increase a health of children and old people.

The Role of Surgical Resection in the Treatment of Newly-Diagnosed Supratentorial Lobar Glioblastoma in Adults (성인에서 천막상부, 두개엽에 위치한 원발성 교모세포종의 치료에서 종양 절제의 역할)

  • Rhee, Jong Joo;Ahn, Jae Sung;Jeon, Sang Ryong;Kim, Jeong Hoon;Ra, Young Shin;Kim, Chang Jin;Lee, Jung Kyo;Kwun, Byung Duk
    • Journal of Korean Neurosurgical Society
    • /
    • v.30 no.sup2
    • /
    • pp.221-227
    • /
    • 2001
  • Objective : The therapeutic impact of tumor resection in glioblastomas is poorly defined and still questionable. Therefore, we conducted the current study to verify the role of tumor resection in the treatment of these highly malignant tumors. Methods : A retrospective study was performed(1990-1999) to compare the treatment results of surgical resection plus radiotherapy(130 patients) with those of stereotactic biopsy plus radiotherapy(19 patients) in glioblastomas. Only adult patients with supratentorial, de novo glioblastoma located in one lobe were included. Survival time/rate was analysed with Kaplan-Meier method, and prognostic variables were obtained from the univariate log-rank test and the multivariate Cox's proportional hazards model. Results : The resection group and the biopsy group did not differ in terms of age, gender, duration of symptoms, presenting symptoms, tumor location, tumor side, tumor size, and the frequency of midline shift. Patients in the biopsy group more often were found to have worse preoperative Karnofsky performance status(KPS)(p=0.001). On univariate analysis, age, KPS, and tumor side were associated with survival(p=0.0053, 0.0001, and 0.0331 respectively). Median survival time and 1-year survival rate were also statistically improved by tumor resection ; resection group - 13 months and 61.2%, and biopsy group - 8 months and 19.7%, respectively(p=0.0001). In patients with midline shift of the tumor, resection was highly effective comparing to biopsy(p=0.0001), but in patients without midline shift, external beam radiation alone was as effective as tumor resection(p=0.0605). Other prognostic variables did not affect survival. On multivariate analysis after variable selection, survival was independently associated with KPS(p=0.001), but not the surgical resection(p=0.2837). Even in biopsy group with midline shift of the tumor, survival rate was not different from that seen after tumor resection(p=0.3505). Conclusions : Radiotherapy alone was as effective as tumor resection plus radiotherapy in patients without midline shift of the tumor. Although there was not statistically significant, tumor resection looked like effective in patients with midline shift. For supratentorial, lobar glioblastoma patients without mass effect of the tumor, biopsy with radiotherapy is one of rational treatment strategies. We consider that tumor resection should be performed in patients with pretreatment midline shift.

  • PDF

Anterior Vaginal Wall Sling for Female Stress Urinary Incontinence (여성 복압성 요실금에서 질전벽슬링수술의 효과)

  • Shin, Hong-Seok;Yoo, Jin-Wook;Jung, Hee-Chang;Park, Tong-Choon
    • Journal of Yeungnam Medical Science
    • /
    • v.18 no.1
    • /
    • pp.59-66
    • /
    • 2001
  • Background: The purpose of this study was to determine the efficacy and safety of the anterior vaginal wall sling in the management of women with stress urinary incontinence. Materials and Methods: From January 1998 to December 1999, 42 patients(31 with genuine stress urinary incontinence and 11 with mixed urinary incontinence, 38 with anatomical incontinence and 4 with intrinsic sphincteric deficiency) underwent anterior vaginal wall sling at Yeungnam University Hospital were studied retrospectively. The mean age was 49.3 years(ranging from 34 to 66 years of age) and the mean follow-up period was 29.4 months(ranging from 16 to 40 months). Intra- and postoperative complication. success rate and patient's satisfaction were evaluated. Results: The mean operation time was 79 minutes(ranging from 65 to 124 minutes) and the mean hospital stay was 5.1 days(ranging from 4 to 10 days). Mean postoperative Foley catheter drainage was 2.1 days(ranging from 1 to 5 days). As a complication. bladder perforation occurred in one patient(2.4%). residual urine sensation developed in seven patients(16.7%), and suprapubic pain was complained in five patients(11.9%), which improved gradually. Vaginal epithelial inclusion cyst occurred in one patient(2.4%) at postoperative 31 months. Four(9.4%) patients with de novo instability were improved by anticholinergics medication. The success rate was 92.9% and 38 patients(90.5%) were satisfied with this procedure. Conclusion: We consider that the anterior vaginal wall sling to be a safe and effective surgical procedure for the treatment of female stress urinary incontinence, but a longer follow-up is necessary to determine long term effect.

  • PDF