• 제목/요약/키워드: PO-DNA

Search Result 73, Processing Time 0.025 seconds

Antibiotics Assay of Doxycycline in Food System using Stripping Voltammetry

  • Ly, Suw Young;Lee, Chang Hyun
    • Journal of the Korean Applied Science and Technology
    • /
    • v.33 no.4
    • /
    • pp.726-733
    • /
    • 2016
  • A voltammetric analysis of doxycycline was developed using DNA immobilized onto a carbon nanotube paste electrode (PE). An anodic peak current was indicated at 0.2 V (versus Ag/AgCl) in a 0.1M $NH_4H_2PO_4$ electrolyte solution. The linear working range of the cyclic and square wave stripping voltammetry was obtained to $1-27ngL^{-1}$ with an accumulation time of 800 s. Final analytical parameters were optimized to be as follows: amplitude, 0.35 V; frequency, 500 Hz; and pH, 5.43. Here detection limit was found to be $0.45ngL^{-1}$, this result can be applied in foods systems and in the biological diagnostics

Isolation and Characterization of an Alkalophilic Cellulolytic Bacterium Pseudomonas sp. (호알칼리성 섬유소분해세균 Pseudomonas sp.의 분리 및 특성)

  • Lim, Sang-Ho;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
    • /
    • v.25 no.1
    • /
    • pp.124-130
    • /
    • 1998
  • An alkalophilic bacterium, the strain AC-711 as a potent producer of alkaline cellulase, was selected among many isolates from soil environments. Morphological, physiological and chemical characteristics of the strain AC-711 suggested that it belongs to the genus Pseudomonas according to the Bergey's Manual of Systematic Bacteriology, however the G+C mol% (54.43) of its chromosomal DNA is lower than the normal values of the genus. The major cell wall fatty acids were determined as 15:0 and 17:0 anteiso. The production of alkaline CMCase by the strain was maximal when grown on the mediun containing 1% carboxymethyl cellulose, 0.1% $KH_2PO_4$, 0.02% $CoCl_2$, 0.02% Tween 80, 0.5% $Na_2CO_3$, 0.8% yeast extract, pH 10.3 at $30^{\circ}C$ for 3 days, and the most of enzyme was excreted into culture mediun.

  • PDF

Study on Measurement of Carcinogen-DNA adducts in exfoliated urothelial cells among workers by 32P-postlabelling methods (근로자의 뇨중 상피세포에서 32P-postlabeling에 의한 발암물질의 DNA adducts측정방법에 대한 연구)

  • Lee, Jin Heon;Roh, Jaehoon;Talaska, Glenn
    • Journal of Korean Society of Occupational and Environmental Hygiene
    • /
    • v.10 no.1
    • /
    • pp.1-17
    • /
    • 2000
  • Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.

  • PDF

BIOACTIVE PEPTIDES DERIVED FROM FOOD PROTEINS AND PREVENTION OF LIFE-STYLE RELATED DISEASES

  • Yoshikawa Masaaki
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
    • /
    • 2001.12a
    • /
    • pp.69-73
    • /
    • 2001
  • Two opioid peptides, YPLDL and YPLDLF, were isolated from enzymatic digests of spinach ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO) and named rubiscolin-5 and -6, respectively. These peptides were selective for delta-receptor and the latter was about 3 times more potent than the former. After oral administration in mice at the dose of 100 mg/kg, rubiscolin-6 showed analgesic activity in tail pinch test. It also stimutated learning performance at the same dose in passive avoidance experiment using step-through apparatus. An immunostimulating peptide, MITLAIPVNKPGR, was isolated from a trypsin digest of soybean protein and named soymetide. Immunostimulating activy of soymetide was mediated by fMLP receptor. Interestingly, after oral administration in rats at a dose of 300 mg/kg (po.), soymetide-4 (MITL) protected alopecia (hair-loss) induced by etoposide, a cancer chemotherapy agent. Stimulation of IL-1 release by the peptide was involved in the mechanism. Ovokinin(2-7), RADHPF, is a vasorelaxing peptide released from ovalbumin by the action of chymotrypsin. It lowered blood pressure of spontaneously hypersensive rats (SHR) after oral administration at a dose of 10 mg/kg. RPLKPW, which was designed by replacing 4 amino acid residues in ovokinin(2-7), exhibited hypotensive activity at a dose of 0.1 mg/kg (po.). This peptides was introduced into 3 homologous sites in soybean beta-conglycinin alpha' subunit by site-directed mutagenesis of the cDNA and expressed in E. coli. The minimum effective dose for hypotensive activity of the genetically modified beta-conglycinin alpha' subunit was 10 mg/kg (po.), which is about 1/200 that of ovalbumin.

  • PDF

Removal Behavior of Biological Nitrogen and Phosphorus and Prediction of Microbial Community Composition with Its Function, in an Anaerobic-Anoxic System form Weak Sewage

  • LEE, JIN WOO;EUI SO CHOI;KYUNG IK GIL;HAN WOONG LEE;SANG HYON LEE;SOO YOOUN LEE;YONG KEUN PARK
    • Journal of Microbiology and Biotechnology
    • /
    • v.11 no.6
    • /
    • pp.994-1001
    • /
    • 2001
  • An easier way of understanding the BNR system was proposed from the study on substrate, nutrient removal tendency, microbial community and its metabolic function by applying the municipal settled sewage. During the anaerobic period, the phosphorus release rate per VFACOD we varied depending on the phosphorus content in the sludge. When the phosphorus content in the sludge was $6\%$ VSS, according to influent VFACOD, the phosphorus release rate and PHA production were $0.35 gPO_4P/gVFACOD$ and 1.0 gPHA/gVFACOD, respectively. The $NO_3N$ requirement for the phosphorus uptake as an electron acceptor was about $0.5 gNO_3N/gPO_4P_{uptake}$ based on the proposed equation with PHA, biomass, production, and the concentration of phosphorus release/uptake. Bacterial-community analysis of the sludge, as determined by FISH and 16SrDNA characterization FISH, revealed that the beta-subclass proteobacteria were the most abundant group ($27.9\%$ of the proteobacteria-specific probe EUB338), and it was likely that representative of the beta-subclass played key roles in activated sludge. The next dominant group found was the gamma-protebacteria ($15.4\%$ of probe EUB338). 16S rDNA clone library analysis showed that the members of${\beta}$- and ${\gamma}$-proteobacteria were also the most abundant groups, and $21.5\%$ (PN2 and PN4) and $15.4\%$ (PN1 and PN5) of total clones were the genera of denitrifying bacteria and PAO, respectively. Prediction of the microbial community composition was made with phosphorus content (Pv, $\%$ P/VSS) in wasted sludge and profiles of COD, PHA, $PO_4P,\;and\;NO_3N$ in an anaerobic-anoxic SBR unit. Generally, the predicted microbial composition based upon metabolic function, i.e., as measured by stoichiometry, is fairly similar to that measure by the unculturable dependent method. In this study, a proposal was made on he microbial community composition that was more easily approached to analyze the reactor behavior.

  • PDF

Genetic Analysis of Sasang Constitution by Amp-FLP Method (Amp-FLP을 이용(利用)한 사상체질(四象體質)의 유전적(遺傳的) 분석(分析) 연구(硏究))

  • Cho, Dong-Wuk;Ahn, Sun-Kyung;Ki, Do-Kyung;Kim, Dae-Won;Ji, Sang-En;Lee, Eui-Ju;Hong, Seok-Chull;Koh, Byung-Hee;Cho, Whang-Sung
    • Journal of Sasang Constitutional Medicine
    • /
    • v.9 no.2
    • /
    • pp.163-173
    • /
    • 1997
  • Amp-FLP is a one of the most frequently used human genetic analysis methods which adopts STR and VNTR typings. In this study, 100 genomic DNA samples of Taeum, Soyang and Soum constitution group were analysed by Amp-FLP method. The allele distribution of three STR loci(TH01, vWA and CSF1PO) and one VNTR locus(MCT118) of each different constitution group were investigated and the allele distribution was statistically evaluated. It was found out that the allele distribution of MCT118 and THO1 loci was not significantly different among different constitutions. However, the allele distribution of vWA showed p-value of 0.02056(Soyang group) and $2.41{\times}10^{-10}$(Soum group) which is much lower than significant level of p-value 0.05. Also, p-value of CSF1PO in Soum group was found out to be $4.65{\times}10^{-17}$. Therefore, it is expected that vWA and CSF1PO loci can be used as an indicator for gnenetic difference of different constiturion if the same result is obtained with sufficient number of samples.

  • PDF

Trends in the rapid detection of infective oral diseases

  • Ran-Yi Jin;Han-gyoul Cho;Seung-Ho Ohk
    • International Journal of Oral Biology
    • /
    • v.48 no.2
    • /
    • pp.9-18
    • /
    • 2023
  • The rapid detection of bacteria in the oral cavity, its species identification, and bacterial count determination are important to diagnose oral diseases caused by pathogenic bacteria. The existing clinical microbial diagnosis methods are time-consuming as they involve observing patients' samples under a microscope or culturing and confirming bacteria using polymerase chain reaction (PCR) kits, making the process complex. Therefore, it is required to analyze the development status of substances and systems that can rapidly detect and analyze pathogenic microorganisms in the oral cavity. With research advancements, a close relationship between oral and systemic diseases has been identified, making it crucial to identify the changes in the oral cavity bacterial composition. Additionally, an early and accurate diagnosis is essential for better prognosis in periodontal disease. However, most periodontal disease-causing pathogens are anaerobic bacteria, which are difficult to identify using conventional bacterial culture methods. Further, the existing PCR method takes a long time to detect and involves complicated stages. Therefore, to address these challenges, the concept of point-of-care (PoC) has emerged, leading to the study and implementation of various chair-side test methods. This study aims to investigate the different PoC diagnostic methods introduced thus far for identifying pathogenic microorganisms in the oral cavity. These are classified into three categories: 1) microbiological tests, 2) microchemical tests, and 3) genetic tests. The microbiological tests are used to determine the presence or absence of representative causative bacteria of periodontal diseases, such as A. actinomycetemcomitans, P. gingivalis, P. intermedia, and T. denticola. However, the quantitative analysis remains impossible, and detecting pathogens other than the specific ones is challenging. The microchemical tests determine the activity of inflammation or disease by measuring the levels of biomarkers present in the oral cavity. Although this diagnostic method is based on increase in the specific biomarkers proportional to inflammation or disease progression in the oral cavity, its commercialization is limited due to low sensitivity and specificity. The genetic tests are based on the concept that differences in disease vulnerability and treatment response are caused by the patient's DNA predisposition. Specifically, the IL-1 gene is used in such tests. PoC diagnostic methods developed to date serve as supplementary diagnostic methods and tools for patient education, in addition to existing diagnostic methods, although they have limitations in diagnosing oral diseases alone. Research on various PoC test methods that can analyze and manage the oral cavity bacterial composition is expected to become more active, aligning with the shift from treatment-oriented to prevention-oriented approaches in healthcare.

Characteristics and breeding of a new oyster mushroom (Pleurotus ostreatus) variety 'Chunhwashim' by using cytoplasmic hybrid (세포질전환 기법에 의한 신품종 느타리 '천화심'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Oh, Youn-Lee
    • Journal of Mushroom
    • /
    • v.13 no.2
    • /
    • pp.129-134
    • /
    • 2015
  • Hybrid strains was selected by crossing between dikaryotic strain ASI 0628 (Dagul) bred from ASI 2596 (Suhan No.3) and ASI 2782(Black pileus mutant) and monokaryotic strain ASI 2344-84. The strain 84 that shown the best cultural characteristics was selected to be a new cytoplasmic hybrid variety and named as 'Chunhwashim'. The 'Chunhwashim' shown incompatibility among them forming the confrontation growth line against parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho). Fruiting body produced about $161.4{\pm}4.8g$ per bottle. And also the individual generation of fruit body is multiple than chunchu2ho as 40.9. The 'Chunhwashim' was cytoplasmic hybrid included hybrid DNA bands of parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho) by Random Amplified Polymorphic DNA (RAPD) primer, and mitochondria DNA band of monokaryotic ASI 2344 (chunchu 2ho) using mitochondria microsatellite DNA marker. This new cytoplasmic hybrid variety 'Chunhwashim' of oyster mushroom is characterized by multiple of individual generation and deeply grey color of pileus.

Isolation, Identification and Optimal Cultrul Condition of Antioxidant Producing Bacterium Isolated from the Marine Sources

  • Kim, Man-Chul;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
    • /
    • 2005.10a
    • /
    • pp.343-346
    • /
    • 2005
  • The isolated strain, SC2-1 was Gram-positive, catalase positive, facultatively anaerobic, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and sodium chloride required for the bacteria growth. The radical scavenging activity of the culture supernatants was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) method. This bacterium was identified based on cellular fatty acids analysis and 16S rDNA sequencing then named Exiguobacterium sp. SC2-1. The optimum culture conditions for production of antioxidant were $25^{\circ}C,$ pH 7.8 and NaCl concentration were 4%. The modified optimal medium compositions were maltose 2.5% (w/v), yeast extract 1.5% (w/v) and $KH_2PO_4$ 0.05% (w/v). Free radical scavenging activity of under optimal culture conditions were 93%.

  • PDF

Theoretical Studies on the Photocycloaddition Reaction of Psoralen with Thymidine

  • Kim, Ja-Hong;Oh, Se-Woung;Lee, Yoon-Sup;Shim, Sang-Chul
    • Bulletin of the Korean Chemical Society
    • /
    • v.8 no.4
    • /
    • pp.298-300
    • /
    • 1987
  • The theoretical studies on the photocycloaddition reaction of 5,7-dimethoxycoumarin and 4',5'-dihydropsoralen with thymidine were carried out as a model for photosensitizing reaction of psoralen with DNA. The results are in reasonable agreement with experimental observations. The photoadducts between dimethoxycoumarin and thymidine were predicted to be $C_{4}$-cycloadducts through the cycloaddition of 3,4-pyrone double bond of dimethoxycoumarin to 5,6 double bond of thymidine. The major photoadduct of 4',5'-dihydropsoralen with thymidine has the anti head-to-head stereochemistry.