• Title/Summary/Keyword: M13 DNA

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Two Cases of MELAS Syndrome Manifesting Variable Clinical Cour (다양한 임상경과를 보인 멜라스(MELAS, mitochondrial encephalopathy, lactic acidosis, and stroke-like episode) 증후군 2례)

  • Choi, Seo Yeol;Lee, Seung-Ho;Myung, Na-Hye;Lee, Young-Seok;Yu, Jeesuk
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.2
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    • pp.102-108
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    • 2016
  • Mitochondrial encephalopathy, lactic acidosis, and stroke-like episode (MELAS) syndrome is one of mitochondrial encephalopathy. As the early clinical manifestations can be variable, it is important to suspect the disease, especially in patients with multiple organ dysfunctions. A boy was diagnosed with epilepsy when he was 9 years old. Two years later, severe headache and blurred vision developed suddenly. On examination, left homonymous hemianopsia was detected with corresponding cerebral parenchymal lesions in right temporo-occipito-parietal areas. MELAS syndrome was confirmed by genetic test, which showed m.3243 A>G mitochondrial DNA mutation. Multivitamins including coenzyme Q10 were added to anticonvulsant. He experienced 4 more events of stroke-like episodes over 5 years, but he is able to perform normal daily activities. A 13-year-old boy was brought to the hospital due to suddenly developed respiratory arrest and asystole associated with pneumonia. Past medical history revealed that he had multiple medical problems such as epilepsy, failure-to-thrive, optic atrophy, and deafness. He has been on valproic acid as an anticonvulsant which was prescribed from local clinic. He recovered after the resuscitation, but his cognition and motor function were severely damaged. He became bed-ridden. He was diagnosed with MELAS syndrome by brain MRI, muscle biopsy, and clinical features. Genetic test did not reveal any mitochondrial gene mutation. Four years later, he expired due to suddenly developed severe metabolic acidosis combined with hyperglycemic hyperosmolar nonketotic coma. The clinical features of MELAS syndrome are variable. Early diagnosis before the presentation to the grave clinical course may be important for the better clinical outcome.

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ANTIBIOTIC SUSCEPTIBILITY OF BACTERIA ISOLATED FROM MAXILLARY SINUSITIS LESION (상악동염 병소 부위에서 세균의 분리 동정 및 항생제 감수성에 대한 연구)

  • Choi, Young-Og;Kim, Su-Gwan;Kim, Hak-Kyun;Kim, Yong-Jong;Choi, Dong-Kook;Kim, Mi-Kwang;Park, Soon-Nang;Kim, Min-Jung;Kook, Joong-Ki
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.5
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    • pp.436-446
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    • 2006
  • The purpose of this study was to isolate and identify the bacteria in chronic maxillary sinusitis (CMS) lesions from 3 patients and to determine the antimicrobial susceptibility of them against 10 antibiotics. One of them was odontogenic origin and the others were non-odontogenic origin. Pus samples were collected by needle aspiration from the lesions and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, CO2, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene (16S rDNA) nucleotide sequencing method. To test the sensitivity of the bacteria isolated from the maxillary sinusitis lesions against seven antibiotics, penicillin G, amoxicillin, tetracycline, ciprofloxacin, cefuroxime, erythromycin, clindamycin, and vancomycin, minimum inhibitory concentration (MIC) was performed using broth dilution assay. Our data showed that enterobacteria such as Enterobacter aerogenes (30%), Klebsiella pneumoniae (25%), and Serratia marcescens (15%) were predominately isolated from the lesion of non-odontogenic CMS of senile patient (70 year old). Streptococcus spp. (40.3%), Actinomyces spp. (27.4%), P. nigrescens, M. micros, and P. anaerobius strains were isolated in the lesion of odontogenic CMS. In the lesion of non-odontogenic CMS, Streptococcus spp. (68.4%), Rothia spp. (13.2%), and Actinomyces sp. (10.5%) were isolated. The susceptibility pattern of 10 antibiotics was determined according to the host of the bacteria strains ratter than the kinds of bacterial species. Even though the number of CMS was limited as three, these results indicate that antibiotic susceptibility test must be accompanied with treatment of CMS. The combined treatment of two or more antibiotics is better than single antibiotic treatment in the presence of multidrug-resistant bacteria in the CMS lesions.

Evaluation and Assessment of Biosafety for Bacillus thuringiensis (Bt)-transgenic Rice: Responses of Daphnia magna Fed on Bt-transgenic Rice Variety (해충저항성 Bacillus thuringiensis (Bt) 벼의 환경위해성 평가: 해충저항성 Bt벼가 물벼룩(Daphnia magna)에 미치는 영향)

  • Oh, Sung-Dug;Shin, Hye-Chul;Sohn, Soo-In;Lee, Ki-Jong;Kim, Hyo-Jin;Ryu, Tae-Hun;Lee, Jang-Yong;Park, Beom-Seok;Kweon, Soon-Jong;Suh, Seok-Cheol;Park, Jong-Sug
    • Journal of Applied Biological Chemistry
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    • v.54 no.4
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    • pp.296-302
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    • 2011
  • Insect-resistant transgenic rice was developed by inserting the mCry1Ac1 a modified gene from the soil bacterium Bacillus thuringiensis (Bt). For biosafety assessment, we studied the effects on survival of cantor Daphnia magna, a commonly used as a model organism in ecotoxicological studies. D. magna fed on Bt rice and its near non-genetically modified (GM) counterparts (Nakdong) grown in the same environment (100% ground rice suspension). The Bt rice was comfirmed to have the insertion of T-DNA and protein expression by the polymerase chain reaction and ELISA analysis. Feeding study showed similar cumulative immobility and abnormal response of D. magna between Bt rice and non-GM counterparts. 48 h-$EC_{50}$ values of Bt rice and non-GM rice showed 4,429 and 2,889 mg/L respectively. The rice no observed effect concentration (NOEC) values for D. magna was suggested 1,000 mg/L. We conclude that the tested Bt-rice and Nakdong similar cumulative immobility for D. magna the widely used model organism. We found out that there is strong possibility that the growth of Bt rice didn't affect to non-target insects.

Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee;Chun, En-Mi;Bae, Myung-Ae;Seu, Young-Bae;Song, Kyung-Sik;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.27-34
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    • 2000
  • The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions

  • Perez, Moises Roberto Vallejo;Contreras, Hugo Ricardo Navarro;Herrera, Jesus A. Sosa;Avila, Jose Pablo Lara;Tobias, Hugo Magdaleno Ramirez;Martinez, Fernando Diaz-Barriga;Ramirez, Rogelio Flores;Vazquez, Angel Gabriel Rodriguez
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.381-392
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    • 2018
  • Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

Roles of CYP1A1 and CYP2E1 Gene Polymorphisms in Oral Submucous Fibrosis

  • Yaming, Punyo;Urs, Aadithya Basavaraj;Saxena, Alpana;Zuberi, Mariyam
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3335-3340
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    • 2016
  • Background: Oral submucous fibrosis (OSF) is a precancerous condition with a 4 to13% malignant transformation rate. Related to the habit of areca nut chewing it is mainly prevalent in South-east Asian countries where the habit of betel quid chewing is frequently practised. On chewing, alkaloids and polyphenols are released which undergo nitrosation and give rise to N-nitrosamines which are cytotoxic agents. CYP450 is a microsomal enzyme group which metabolizes various endogenous and exogenous chemicals including those released by areca nut chewing. CYP1A1 plays a central role in metabolic activation of these xenobiotics, whereas CYP2E1 metabolizes nitrosamines and tannins. Polymorphisms in genes that code for these enzymes may alter their expression or function and may therefore affect an individuals susceptibility regarding OSF and oral cancer. The present study was therefore undertaken to investigate the association of polymorphisms in CYP1A1 m2 and CYP2E1 (RsaI/PstI) sites with risk of OSF among areca nut chewers in the Northern India population. A total of 95 histopathologically confirmed cases of OSF with history of areca nut chewing not less than 1 year and 80, age and sex matched controls without any clinical signs and symptoms of OSF with areca nut chewing habit not less than 1 year were enrolled. DNA was extracted from peripheral blood samples and polymorphisms were analyzed by PCR-RFLP method. Gene polymorphism of CYP1A1 at NcoI site was observed to be significantly higher (p = 0.016) in cases of OSF when compared to controls. Association of CYP1A1 gene polymorphism at NcoI site and the risk of OSF (Odd's Ratio = 2.275) was also observed to be significant. However, no such association was observed for the CYP2E1 gene polymorphism (Odd's Ratio = 0.815). Our results suggest that the CYP1A1 gene polymorphism at the NcoI site confers an increased risk for OSF.

Protection of Infection and Eradication Activity of Culture Product by Pediococcus pentosaceus CBT SL4 Showing Antimicrobial Activity against Helicobacter pylori (Helicobacter pylori에 대한 항균활성을 나타내는 Pediococcus pentosaceus CBT SL4 배양물의 감염방어 및 제균활성)

  • Hong, Un-Pyo;Chung, Myung-June;Kim, Soo-Dong;Oh, Eun-Taex;So, Jae-Seong;Chung, Chung-Il
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.779-783
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    • 2004
  • New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.

Purification of Methioninase from Pseudomonas putida and Its Effect on the Uptake of ^11C-Methionine in Vivo. (Pseudomonas putida 유래 Methioninase의 정제 및 생체내 ^11C-Methionine 섭취에 미치는 영향)

  • 변상성;박귀근
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.377-382
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    • 2003
  • Purification of methioninase resulted in a yield of 69%, and SDS-PAGE analysis of the purified product revealed a single band of approximately 43 kDa in molecular weight. in vitro experiments with cancer cells incubated in methionine-free media demonstrated an increase in $^{11}$ C-methionine uptake to 25.8$\pm$1.1% at 6 hr, 31.8$\pm$0.8% at 24 hr, and 62.2$\pm$0.6% at 48hr, compared to controls. Treatment of the cancer cells with purified methioninase showed no decrease in survival after a 2 hr incubation with 0.01 U/ml, but survival of RR1022 cells decreased 30% after 24 to 48 hr incubation. SKOV-3 cells showed a 5% and 14% decrease in survival with 0.1 and 1 U/ml methioninase after 24 hr. After 48hr survival decreased 15% and 24% with 0.1 and 1 U/ml methioninase. Measurements of $^{11}$ C-methionine uptake in RR1022 cells demonstrated no change at 2 hr, but a 13.7$\pm$4.7% and 40.7$\pm$2.6% increase in uptake at 24 and 48 hr, respectively. SKOV-3 cells also showed no change at 2 hr, but had a 17.7$\pm$7.2% and 38.9$\pm$4.9% increase in $^{11}$ C-methionine uptake after 24 hr and 48 hr treatment with methioninase, respectively. $^{11}$ C-methionine PET imaging revealed clear visualization of both the tumors and contralateral infectious lesions. Administration of rMET appeared to result in a slight increase in tumor:nontumor contrast on $^{11}$ C-methionine PET images. Injection of purified methioninase also produced PET images where tumor uptake was higher than that of infectious lesions.

Assessment of the level and identification of airborne molds by the type of water damage in housing in Korea (국내 주택에서 물 피해 유형에 따른 부유곰팡이 농도 수준 평가 및 동정 분석)

  • Lee, Ju Yeong;Hwang, Eun Seol;Lee, Jeong-Sub;Kwon, Myunghee;Chung, Hyen Mi;Seo, SungChul
    • Journal of odor and indoor environment
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    • v.17 no.4
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    • pp.355-361
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    • 2018
  • Mold grows more easily when humidity is higher in indoor spaces, and as such is found more often on wetted areas in housing such as walls, toilets, kitchens, and poorly managed spaces. However, there have been few studies that have specifically assessed the level of mold in the indoor spaces of water-damaged housing in the Republic of Korea. We investigated the levels of airborne mold according to the characteristics of water damage types and explored the correlation between the distribution of mold genera and the characteristics of households. Samplings were performed from January 2016 to June 2018 in 97 housing units with water leakage or condensation, or a history of flooding, and in 61 general housing units in the metropolitan and Busan area, respectively. Airborne mold was collected on MEA (Malt extract agar) at flow rate of 100 L/min for 1 min. After collection, the samples were incubated at $25^{\circ}C$ for 120 hours. The cultured samples were counted and corrected using a positive hole conversion table. The samples were then analyzed by single colony culture, DNA extraction, gene amplification, and sequencing. By type of housing, concentrations of airborne mold were highest in flooded housing, followed by water-leaked or highly condensed housings, and then general housing. In more than 50% of water-damaged housing, the level of airborne mold exceeded the guideline of Korea's Ministry of Environment ($500CFU/m^3$). Of particular concern was the fact that the I/O ratio of water-damaged housing was greater than 1, which could indicate that mold damage may occur indoors. The distribution patterns of the fungal species were as follows: Penicillium spp., Cladosporium spp. (14%), Aspergillus spp. (13%) and Alternaria spp. (3%), but significant differences of their levels in indoor spaces were not found. Our findings indicate that high levels of mold damage were found in housing with water damage, and Aspergillus flavus and Penicillium brevicompactum were more dominant in housing with high water activity. Comprehensive management of flooded or water-damaged housing is necessary to reduce fungal exposure.

Effects of 2-Deoxy-D-Glucose on Metabolic Status, Proliferative Capacity and Growth Rate of FSall Tumor: Observations made by In Vivo $^{31}P$-Nuclear Magnetic Resonance Spectroscopy and Flow Cytometry (2-DDG가 FSa II 종양의 성장속도와 증식 능력, 신진대사에 미치는 영향 ; $^{31}P$-자기공명 분광기와 유세포 분석기를 이용한 연구)

  • Chang Hyesook;Choi Eun Kyung;Cho Jeong Gill;Lim Tee-Hwon;Lee Tae-Keun;Yi Yun;Cho Young Joo;Kim Gon Sup
    • Radiation Oncology Journal
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    • v.9 no.1
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    • pp.1-6
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    • 1991
  • The effect of 2-deoxy-d-glucose (2-DDG) on $C_3H$ mouse fibrosarcoma(FSall) was studied. Metabolic status, especially for energy metabolism, was studied using in vivo $^{31}P$-MRS, proliferative capacity was observed on flow cytometry(FC) and growth rate was measured after transplantation of $10^6$ viable tumor cells in the dorsum of foot of $C_3Hf/Sed$ mice. One gram of 2-DDG Per kg of body weight was injected intraperitoneally on 12th day of implantation. Average tumor size on 12th day of implantion was $250mm^3$. Growth rate of Fsall tumor was measured by tumor doubling time and slope on semilog plot. After 2-DDG injection, growth rate slowed down. Tumor doubling time between tumor age 5-12 days was 0.84 days with slope 0.828 and tumor doubling time between tumor age 13-28 days was 3.2 days with slope 0.218 in control group. After 2-DDG injection, tumor doubling time was elongated to 5.1 days with slope 0.136. The effect of 2-DDG studied in vivo $^{31}P$-MRS suggested that the increase of phosphomonoester (PME) and inorganic phosphate (Pi) by increasing size of tumor, slowed down after 2-DDG injection. Flow cytometry showed significantly increased S-phase and $G_2+M$ phase fraction suggesting increased proliferative capacity of tumor cells in the presence of 2-DDG. Authors observed an interesting effect of 2-DDG on FSall tumor and attempt to utilize as an adjunct for radiotherapy.

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