• Title/Summary/Keyword: Frequency specificity

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The Photoheterotrophic Growth of Bacteriochlorophyll Synthase-Deficient Mutant of Rhodobacter sphaeroides Is Restored by I44F Mutant Chlorophyll Synthase of Synechocystis sp. PCC 6803

  • Kim, Eui-Jin;Kim, Hyeonjun;Lee, Jeong K.
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.959-966
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    • 2016
  • Chlorophyll synthase (ChlG) and bacteriochlorophyll synthase (BchG) have a high degree of substrate specificity. The BchG mutant of Rhodobacter sphaeroides, BG1 strain, is photosynthetically incompetent. When BG1 harboring chlG of Synechocystis sp. PCC 6803 was cultured photoheterotrophically, colonies arose at a frequency of approximately 10-8. All the suppressor mutants were determined to have the same mutational change, ChlGI44F. The mutated enzyme ChlGI44F showed BchG activity. Remarkably, BchGF28I, which has the substitution of F at the corresponding 28th residue to I, showed ChlG activity. The Km values of ChlGI44F and BchGF28I for their original substrates, chlorophyllide (Chlide) a and bacteriochlorophyllide (Bchlide) a, respectively, were not affected by the mutations, but the Km values of ChlGI44F and BchGF28I for the new substrates Bchlide a and Chlide a, respectively, were more than 10-fold larger than those for their original substrates, suggesting the lower affinities for new substrates. Taken together, I44 and F28 are important for the substrate specificities of ChlG and BchG, respectively. The BchG activity of ChlGI44F and the ChlG activity of BchGF28I further suggest that ChlG and BchG are evolutionarily related enzymes.

DEVELOPMENT OF ANIMAL CELL EXPRESSION SYSTEM WHICH CONFERS POSITION-INDEPENDENT AND ENHANCED FOREIGN GENE EXPRESSION

  • Yoon, Yeup;Kim, Jong-Mook;Kim, Jung-seob;Oh, Sun-Mo;Kim, Jong-Il;Yoon, Jae-Seung;Baek, Kwang-Hee
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.178-181
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    • 2000
  • In order to develop the novel gene expression system, we introduced new control elements which could influence the foreign gene expression in animal cells. When the foreign genes are introduced into the genome of higher eukaryotic cells, the expressions from these integrated genes are often low and can vary greatly depending on the positions of the integration sites due to the complex nature of the chromatin structures (1). First we screened the various DNA sequence elements which can function as an insulator of gene expression from these position effects and can cooperate with the SV40 enhancer/promoter. Among the several DNA elements from the various sources, we identified the particular DNA element which confers the increased frequency of the positive colonies, assayed by the reporter gene from stable selections indicating significantly reduced position effects. This element also showed the several fold-increased expression level as well as the copy-number dependent expression with host cell specificity. Second we modified the transcription termination element where we introduced the specific terminator in combination with SV40 polyA signal. This modified terminator showed the increased efficiency and the level of the gene expression. By combining these two elements, we made the animal cell expression system and tested successfully for the recombinant protein productions of TGF ${\beta}$-soluble receptor, Antithrombin III, and single chain Pro-Urokinase. [Supported by grants from MOCIE]

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Massive Parallel Sequencing for Diagnostic Genetic Testing of BRCA Genes - a Single Center Experience

  • Ermolenko, Natalya A;Boyarskikh, Uljana A;Kechin, Andrey A;Mazitova, Alexandra M;Khrapov, Evgeny A;Petrova, Valentina D;Lazarev, Alexandr F;Kushlinskii, Nikolay E;Filipenko, Maxim L
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7935-7941
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    • 2015
  • The aim of this study was to implement massive parallel sequencing (MPS) technology in clinical genetics testing. We developed and tested an amplicon-based method for resequencing the BRCA1 and BRCA2 genes on an Illumina MiSeq to identify disease-causing mutations in patients with hereditary breast or ovarian cancer (HBOC). The coding regions of BRCA1 and BRCA2 were resequenced in 96 HBOC patient DNA samples obtained from different sample types: peripheral blood leukocytes, whole blood drops dried on paper, and buccal wash epithelia. A total of 16 random DNA samples were characterized using standard Sanger sequencing and applied to optimize the variant calling process and evaluate the accuracy of the MPS-method. The best bioinformatics workflow included the filtration of variants using GATK with the following cut-offs: variant frequency >14%, coverage ($>25{\times}$) and presence in both the forward and reverse reads. The MPS method had 100% sensitivity and 94.4% specificity. Similar accuracy levels were achieved for DNA obtained from the different sample types. The workflow presented herein requires low amounts of DNA samples (170 ng) and is cost-effective due to the elimination of DNA and PCR product normalization steps.

CCNA1 Promoter Methylation: a Potential Marker for Grading Papanicolaou Smear Cervical Squamous Intraepithelial Lesions

  • Chujan, Suthipong;Kitkumthorn, Nakarin;Siriangkul, Sumalee;Mutirangura, Apiwat
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7971-7975
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    • 2014
  • Background: From our previous study, we established that cyclin A1 (CCNA1) promoter methylation is strongly correlated with multistep progression of HPV-associated cervical cancer, suggesting potential use as a diagnostic maker of disease. Objectives: The purpose of the present study was to assess the prevalence of CCNA1 promoter methylation in residual cervical cells isolated from liquid-based cytology that underwent hrHPV DNA screening for cervical cancer, and then to evaluate this marker for diagnostic accuracy using parameters like sensitivity, specificity, predictive values and likelihood ratio. Methods: In this retrospective study, histopathology was used as the gold standard method with specimens separated into the following groups: negative (n=31), low-grade squamous intraepithelial lesions (LSIL, n=34) and high-grade squamous intraepithelial lesions or worse (HSIL+, n=32). The hrHPV was detected by Hybrid Capture 2 (HC2) and CCNA1 promoter methylation was examined by CCNA1 duplex methylation specific PCR. Results: The results showed the frequencies of CCNA1 promoter methylation were 0%, 5.88% and 83.33%, while the percentages of hrHPV were 66.67%, 82.35% and 100% in the negative, LSIL and HSIL+ groups, respectively. Although hrHPV infection showed high frequency in all three groups, it could not differentiate between the different groups and grades of precancerous lesions. In contrast, CCNA1 promoter methylation clearly distinguished between negative/LSIL and HSIL+, with high levels of all statistic parameters. Conclusion: CCNA1 promoter methylation is a potential marker for distinguishing between histologic negative/LSIL and HSIL+using cervical cytology samples.

A Clinical Study on Surgical Thyroid Nodules (외과적 갑상선 결절에 대한 임상적 고찰)

  • Park Hyeon-Jung;Moon Sang-Eun
    • Korean Journal of Head & Neck Oncology
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    • v.9 no.2
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    • pp.234-243
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    • 1993
  • The thyroid nodules are the most common endocrine disease requiring surgical management. Up to date, various diagnostic techniques and surgical management have been developed. Authors analysed 2285 cases of thyroid nodules who were treated at Department of Surgery, Pusan National University for the duration of 10 years from January 1980 to December 1989 and the results obtained were summerized as follows: 1) Patients were composed of 1727 cases(75.8%) of benign nodule and 558 cases(24.5%) of malignant nodule. Benign nodule was prevalent in forth and fifth decade comparing with malignant nodule was sixth and fifth decade. The sexual distribution revealed female preponderance with 1:10.8 in benign nodule and 1:9.3 in malignant nodule. 2) The histopathologic classfication of benign nodule in decreasing order of frequency were follicular adenoma 1009 cases(54.8%), adenomatous goiter 573 cases(33.3%), simple cyst 65 cases(3.8%), and Hashimoto's thyroiditis 52 cases(3.0%). The malignant disease were papillary adenocarcinoma 460 cases(82.4%), follicular adenocarcinoma 69 cases(12.4%), undifferentiated carcinoma 13 cases (2.0%), and medullary carcinoma 7 cases(1.0%). 3) Fine needle aspiration cytology was performed in 1758 cases and it showed 80.5% of sensitivity, 96.5% of specificity, 19.5% of false negative and 80% of accuracy. 4) The location of nodule was 87.7%, in unilateral, 12.2% in bilateral. 5) On the radioiodine scanning, the incidence of benign nodule with cold nodule was 83.9% and the incidence of benign nodule with hot nodule was 7.5%. The incidence of thyroid carcinoma with cold nodule was 88.3% and the incidence of thyroid carcinoma with hot nodule was 4.6%. 6) Most cases of benign nodules were treated with lobectomy 82.6%, subtotal thyroidectomy 10.1% and subtotal lobectomy 3.0%. Malignant nodules were treated with lobectomy and/or isthmusectomy 37.6%, total thyroidectomy 17.7%, subtotal thyroidectomy 15.4%, and any thyroidectomy and neck dissection 13.5%. 7) Postoperative complications were developed in 3.8% of benign nodules and 13.8% of malignant nodules.

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Improved Diagnostic Accuracy in Characterization of Adnexal Masses by Detection of Choline Peak Using 1H MR Spectroscopy in Comparison to Internal Reference at 3 Tesla

  • Malek, Mahrooz;Pourashraf, Maryam;Gilani, Mitra Modares;Gity, Masoumeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.5085-5088
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    • 2015
  • Background: The aim of this study was to assess the role of the presence of a choline peak in 3 Tesla 1H magnetic resonance spectroscopy (MRS) for differentiating benign from malignant adnexal masses. Materials and Methods: A total of 46 adnexal masses (23 malignant and 23 benign) underwent 1H MRS study prior to surgery to assess the presence of choline peak. Results: A choline peak was detected in 16 malignant masses (69.5%) and was absent in the other 7 (30.5%). A choline peak was only detected in 6 (26%) of the benign adnexal masses. The presence of an MRS choline peak had a sensitivity of 69.5%, a specificity of 74%, a positive predictive value (PPV) of 72.7%, and a negative predictive value (NPV) of 71% for diagnosing malignant adnexal masses. A significant difference between the frequency of mean choline peaks in benign and malignant adnexal masses was observed (P value < 0.01). Conclusions: A 1H MRS choline peak is seen in malignant adnexal masses more frequently than the benign masses, and may be helpful for diagnosing malignant adnexal masses.

A Study on the Criteria Appropriateness of Experimental Materials & Facilities in Home Economics Education of Secondary School (중등학교 가정과 실험학습 교구.설비기준의 적정성에 관한 연구)

  • 강혜숙;윤인경
    • Journal of Korean Home Economics Education Association
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    • v.5 no.1
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    • pp.61-85
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    • 1993
  • Because of the specificity of subjects belong to middle school curriculum of Home Economics Education of function training is very important. but it pointed out that the existing standards are out-dated, and because of it , it incongruent to our realities. Therefore, whenever the curriculum o education and the national textbook are reorgrnized periodically, the reform of standards of educational materials & facilities must be followed by it. But in my opinion, in korean middle school, especially in Home Economics Education, educational facilities are not reformed at a proper time. Therefore to enhance the qualities of experimentation/practice of Home Economics Education, in the first place, the preparation for the given conditions of standards of educational materials & facilities are needed. But the educational facilities of Home Economics Education have many problem not only in the qualitative aspect but also in the quantitative aspect. And in addition to it, lost of educational facilities which cannot match contents of national textbook of Home Economics Education are exist. Therefore this thesis aims at examining comprehensively the problem of experimentation /practice of experimental materials & facilities of Home Economics Education connect with contents of Home Economics Education, holding several problems of experimentation/practice of Home Economics Education in middle school, grouping ; the principles and ways concerning the innovation experimental materials & facilities of Home Economics Education by which many teachers and students make sure of more efficient teaching/studying, and obtaining basic data which guarantee the ability of innovation concerning problems of the education of experimentation/practice of Home Economics Education. Method are : 1)grasp our educational realities with my original(question). 2)Integrate/analyse results of 1) 3)check the frequency, percentage with methods of X$^2$-test, T-test, and ‘One-way Analysis of Variances’. 4)suggest my own alternative proposal.

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Research for general regulation acupoints prescription's in (<침금방(千金方)>의 침구처방용혈(鍼灸處方用穴)의 일반규율(一般規律) 연구(硏究))

  • Kim, Yun-Jin;Kim, Jae-Hyo;Li, Zhong-Ren;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.22 no.4
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    • pp.161-168
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    • 2005
  • Objective : <千金方(Qianjinfang)> which documents a great amount of prescription of Acupuncture and moxibustion takes an important role during the development of acupuncture and moxibustion. Thus, it have given rise us to the research-situation of Tang Dynasty China of previous period. This study was to outline the rules of choosing acupoints as well as the prescriptions of contemporary times in reference with <千金方(Qianjinfang)>, and to discuss the conception of acupuncture and moxibustion in <千金方(Qianjinfang)>. Method : Based on <千金方(Qianjinfang)>, the parameters were categorized to a certain scope, syndromes in different type were classified. Also numbers of acupoints, route of meridians locations, utilities of special acupoints even prescription methods were statistically analyzed in reference with <千金方(Qianjinfang)>. Results & Conclusion : Acupuncture prescription in <千金方(Qianjinfang)> was mostly presented by single acupoint and this was basic prescription of its rule of choosing acupoints. Choosing acupoint for the majority of various diseases started from choosing meridians with the disorder, but no rules for choosing acupoints was clear. There was basically various methods in choosing acupoints, but little were used by Biao-li meridian choosing method (表裏經配穴). In the high frequency of use, wu-shu acupoints was higher used than other specific acupoints (特定穴). While wu-shu acupoints treated the major parts of the diseases, shu-mo acupoints were strictly used on system. Consequently, The theoretical basis originating the choosing rules of the acupoints and prescriptions was shown in <手金方(Qianjinfang)> as representative clinical reference, through which it was valuable to analyze the prescription rule and specificity of the acupoints.

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Sleep Apnea Detection Using a Piezo Snoring Sensor: A Pilot study (코골이용 압전센서를 이용한 수면무호흡 검출에 관한 예비 연구)

  • Urtnasan, Erdenebayar;Lee, Hyo-Ki;Kim, Hojoong;Lee, Kyoung-Joung
    • Journal of Biomedical Engineering Research
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    • v.35 no.4
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    • pp.75-80
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    • 2014
  • This paper proposed a method that can automatically classify sleep apnea by using features extracted from pulse rate variability(PRV) signals induced from piezo snoring sensor for patients with obstructive sleep apnea(OSA). We have extracted eight features(NN, SDNN, RMSSD, NN10, NN50, LF, HF and LF/HF ratio) based on time and frequency analyses of PRV. Sleep apnea was classified by a linear discriminant analysis(LDA). A performance was evaluated using snore recordings from 13 patients with OSA (ages: $54.5{\pm}10.5$ years, body mass index: $26.3{\pm}2.5kg/m^2$, apnea-hypopnea index: $19.2{\pm}6.0/h$). The sensitivity and specificity were $78.9{\pm}0.9%$ and $78.9{\pm}0.9%$ for training set and $77.7{\pm}10.9%$ and $79.0{\pm}2.8%$ for test set, respectively. Our study demonstrated the feasibility of implementing a piezo snoring sensor based on a portable device as a simple and cost-effective solution for contributing to the OSA screening.

A novel F-box protein with leucine-rich repeats affects defecation frequency and daumone response in Caenorhabditis elegans

  • Kim, Sung-Moon;Jang, Sang-Ho;Son, Na-Rae;Han, Ching-Tack;Min, Kwan-Sik;Lee, Hak-Kyo;Hwang, Sue-Yun
    • Animal cells and systems
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    • v.16 no.4
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    • pp.280-288
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    • 2012
  • Targeted degradation of proteins through ubiquitin-mediated proteolysis is an important control mechanism in various cellular processes. The process of ubiquitin conjugation is achieved by three enzyme complexes, among which the ubiquitin ligase complex (E3) is in charge of substrate specificity. The SCF (SKP1-CUL1-F-box) family portrays the largest and the most characterized member of the E3 ligases. For each SCF complex, the ubiquitination target is recognized by the F-box protein subunit, which interacts with the substrate through a unique C-terminal domain. We have characterized a novel F-box protein CFL-1 that represents a single LRR-type F-box (FBXL) in the Caenorhabditis elegans genome. CFL-1 is highly homologous to FBXL20 and FBXL2 of mammals, which are known to regulate synaptic vesicle release and cell cycle, respectively. A green fluorescence protein (GFP)-reporter gene fused to the cfl-1 promoter showed restricted expression around the amphid and the anus. Modulation of CFL-1 activity by RNAi affected the time interval between defecations. RNAi-treated worms also exhibited reduced tendency to form dauer when exposed to daumone. The potential involvement of CFL-1 in the control of defecation and pheromone response adds to the ever expanding list of cellular processes controlled by ubiquitin-mediated proteolysis in C. elegans. We suggest that CFL-1, as a single LRR-type F-box protein in C. elegans, may portray a prototype gene exerting diverse functions that are allocated among multiple FBXLs in higher organisms.