• Title/Summary/Keyword: Gene testing

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Comparing In Vitro and In Vivo Genomic Profiles Specific to Liver Toxicity Induced by Thioacetamide

  • Kang, Jin-Seok;Jeong, Youn-Kyoung;Shin, Ji-He;Suh, Soo-Kyung;Kim, Joo-Hwan;Lee, Eun-Mi;Kim, Seung-Hee;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.252-260
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    • 2007
  • As it is needed to assay possible feasibility of extrapolation between in vivo and in vitro systems and to develop a new in vitro method for toxicity testing, we investigated global gene expression from both animal and cell line treated with thioacetamide (TAA) and compared between in vivo and in vitro genomic profiles. For in vivo study, mice were orally treated with TAA and sacrificed at 6 and 24 h. For in vitro study, TAA was administered to a mouse hepatic cell line, BNL CL.2 and sampling was carried out at 6 and 24 h. Hepatotoxicity was assessed by analyzing hepatic enzymes and histopathological examination (in vivo) or lactate dehydrogenase (LDH) assay and morphological examination (in vitro). Global gene expression was assessed using microarray. In high dose TAA-treated group, there was centrilobular necrosis (in vivo) and cellular toxicity with an elevation of LDH (in vitro) at 24 h. Statistical analysis of global gene expression identified that there were similar numbers of altered genes found between in vivo and in vitro at each time points. Pathway analysis identified several common pathways existed between in vivo and in vitro system such as glutathione metabolism, bile acid biosynthesis, nitrogen metabolism, butanoate metabolism for hepatotoxicty caused by TAA. Our results suggest it may be feasible to develop toxicogenomics biomarkers by comparing in vivo and in vitro genomic profiles specific to TAA for application to prediction of liver toxicity.

Virulence Genes of Helicobacter pylori in Gastritis, Peptic Ulcer and Gastric Cancer in Laos

  • Vannarath, Sengdao;Vilaichone, Ratha-Korn;Rasachak, Bouachanh;Mairiang, Pisaln;Yamaoka, Yoshio;Shiota, Seiji;Binh, Tran Thanh;Mahachai, Varocha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.9027-9031
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    • 2014
  • Background: Helicobacter pylori (H. pylori) infection is an established cause of peptic ulcers and gastric cancer. The aim of this study was to identify H. pylori genotypes and to examine their associations with geographical regions and gastritis, peptic ulcers and gastric cancer in Laos. Materials and Methods: A total of 329 Lao dyspeptic patients who underwent gastroscopy at Mahosot Hospital, Vientiane, Laos during December 2010 - March 2012 were enrolled. Two biopsy specimens (one each from the antrum and corpus) were obtained for CLO testing and only CLO test-positive gastric tissue were used to extract DNA. PCR and sequencing were identified for variants of the cagA and vacA genotypes. Results: Some 119 Laos patients (36.2%) were found to be infected with H. pylori including 83 with gastritis, 13 with gastric ulcers (GU), 20 with duodenal ulcers (DU) and 3 with gastric cancer. cagA was detected in 99.2%. East-Asian-type cagA (62%) and vacA s1c (64.7%) were predominant genotypes in Laos. vacA s1c-m1b was significantly higher in GU than gastritis (53.8% vs. 24.1%; P-value=0.04) whereas vacA s1a-m2 was significantly higher in DU than gastritis (40.0% vs. 16.9%; P-value=0.03). East-Asian-type cagA and vacA s1c were significantly higher in highland than lowland Lao (100% vs. 55.8%; P-value=0.001 and 88.2% vs. 61.5%, P-value=0.03 respectively). Conclusions: H. pylori is a common infection in Laos, as in other countries in Southeast Asia. The cagA gene was demonstrated in nearly all Laos patients, cagA and vacA genotypes being possible important factors in explaining H. pylori infection and disease outcomes in Laos.

Integrated RT-PCR Microdevice with an Immunochromatographic Strip for Colorimetric Influenza H1N1 virus detection

  • Heo, Hyun Young;Kim, Yong Tae;Chen, Yuchao;Choi, Jong Young;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.273-273
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    • 2013
  • Recently, Point-of-care (POC) testing microdevices enable to do the patient monitoring, drug screening, pathogen detection in the outside of hospital. Immunochromatographic strip (ICS) is one of the diagnostic technologies which are widely applied to POC detection. Relatively low cost, simplicity to use, easy interpretations of the diagnostic results and high stability under any circumstances are representative advantages of POC diagnosis. It would provide colorimetric results more conveniently, if the genetic analysis microsystem incorporates the ICS as a detector part. In this work, we develop a reverse transcriptase-polymerase chain reaction (RT-PCR) microfluidic device integrated with a ROSGENE strip for colorimetric influenza H1N1 virus detection. The integrated RT-PCR- ROSGENE device is consist of four functional units which are a pneumatic micropump for sample loading, 2 ${\mu}L$ volume RT-PCR chamber for target gene amplification, a resistance temperature detector (RTD) electrode for temperature control, and a ROSGENE strip for target gene detection. The device was fabricated by combining four layers: First wafer is for RTD microfabrication, the second wafer is for PCR chamber at the bottom and micropump channel on the top, the third is the monolithic PDMS, and the fourth is the manifold for micropump operation. The RT-PCR was performed with subtype specific forward and reverse primers which were labeled with Texas-red, serving as a fluorescent hapten. A biotin-dUTP was used to insert biotin moieties in the PCR amplicons, during the RT-PCR. The RT-PCR amplicons were loaded in the sample application area, and they were conjugated with Au NP-labeled hapten-antibody. The test band embedded with streptavidins captures the biotin labeled amplicons and we can see violet colorimetric signals if the target gene was amplified with the control line. The off-chip RT-PCR amplicons of the influenza H1N1 virus were analyzed with a ROSGENE strip in comparison with an agarose gel electrophoresis. The intensities of test line was proportional to the template quantity and the detection sensitivity of the strip was better than that of the agarose gel. The test band of the ROSGENE strip could be observed with only 10 copies of a RNA template by the naked eyes. For the on-chip RT-PCR-ROSGENE experiments, a RT-PCR cocktail was injected into the chamber from the inlet reservoir to the waste outlet by the micro-pump actuation. After filling without bubbles inside the chamber, a RT-PCR thermal cycling was executed for 2 hours with all the microvalves closed to isolate the PCR chamber. After thermal cycling, the RT-PCR product was delivered to the attached ROSGENE strip through the outlet reservoir. After dropping 40 ${\mu}L$ of an eluant buffer at the end of the strip, the violet test line was detected as a H1N1 virus indicator, while the negative experiment only revealed a control line and while the positive experiment a control and a test line was appeared.

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Investigation of Porcine Melanocortin-4 Receptor (MC4R) Polymorphism on Economic Traits (돼지 melanocortin-4 receptor (MC4R) 유전자의 경제형질과의 연관성에 관한 연구)

  • Kim Kwan-Suk;Shin Hee Young;Lee Joong-Jae;Hong Sung-Kwang;Choi Bong-Hwan;Kim Tae-Hun;Lee Hak-Kyo;Cho Byung-Wook
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.968-971
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    • 2005
  • The Melanocortin-4 receptor gene (MC4R) regulates the energy balance and the genetic basis of obesity. A polymorphism in the porcine melanocortin-4 receptor has previously shown to be associated with growth, fat deposition and feed intake. In this study, the polymorphism of the gene was studied in several pig breeds of Duroc, Landrace, Berkshire, and Yorkshire. The results showed that the frequencies of MC4R genotype varied among those breeds. Association analyses were also performed between the MC4R polymorphism and average daily gain, feed conversion ratio, backfat thickness and lean percentage phenotypes. The results strongly support that the MC4R polymorphism can be used DNA marker selection indicator for economically important traits for pig breeding program in Korea.

Population genetic structure of Sedum polytrichoides (Crassulaceae): Insights into barriers to gene flow (바위채송화(돌나물과)집단의 유전적 구조: 유전자 이동과 물리적 장벽에 관한 통찰)

  • Chung, Mi Yoon;Lopez-Pujol, Jordi;Chung, Myong Gi
    • Korean Journal of Plant Taxonomy
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    • v.46 no.4
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    • pp.361-370
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    • 2016
  • An area comprising Juwangsan National Park and its adjacent mountains (southeastern Korean Peninsula) is a good model system for testing the effects of physical barriers to gene flows in plant populations. We predicted that plant species consisting of isolated populations are genetically more differentiated than those that are rather continuously distributed. Most populations of Sedum polytrichoides occur in four isolated valleys, and we assessed the genetic variability and structures using twelve allozyme loci in ten populations. We also compared the present results to earlier findings pertaining to the two co-occurring herbs Hylotelephium ussuriense (${\equiv}$ Sedum ussuriense) (growing only in the four isolated valleys) and S. kamtschaticum (rather continuously distributed). We found moderate levels of within-population genetic variation in S. polytrichoides ($H_{e}=0.112$). Estimates of among-population divergence in S. polytrichoides were also moderate ($F_{ST}=0.250$) and, as expected, very similar to that of H. ussuriense (0.261) but considerably higher than the variation in S. kamtschaticum (0.165). An analysis of molecular variance (AMOVA) revealed that S. polytrichoides and H. ussuriense had higher percentages of among-valley variation (19% each) than S. kamtschaticum (4%). Most of this variation, as also indicated by the STRUCTURE program, was due to differences in genetic profiles between the two central valleys. We concluded that the genetic differences observed between species (S. kamtschaticum vs. S. polytrichoides and H. ussuriense) are mainly due to differences in their distribution within the study area.

Molecular Detection of Virulence Factors in Carbapenem-Resistant Pseudomonas aeruginosa Isolated from a Tertiary Hospital in Daejeon (대전지역의 3차 병원에서 분리된 Carbapenem 내성 Pseudomonas aeruginosa의 병독성 인자 검출)

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.301-308
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    • 2019
  • The emergence and spread of multidrug resistant (MDR) Pseudomonas aeruginosa is a critical problem worldwide. The pathogenesis of P. aeruginosa is due partly to the production of several cell-associated and extracellular virulence factors. This study examined the distribution of virulence factors and antimicrobial resistance patterns of carbapenem-resistant P. aeruginosa (CRPA) isolated from a tertiary hospital in Daejeon, Korea. Antimicrobial susceptibility testing was performed using the disk diffusion method, and PCR and DNA sequencing were performed to determine for the presence of virulence genes. In addition, the sequence type (ST) of MDR P. aeruginosa was investigated by multilocus sequence typing (MLST). Among 32 CRPA isolates, 14 (43.8%) were MDR and the major ST was ST235 (10 isolates, 71.4%). All isolates were positive for the presence of virulence genes and the most prevalent virulence genes were toxA, plcN, and phzM (100%). All isolates carried at least eight or more different virulence genes and nine (28.1%) isolates had 15 virulence genes. The presence of the exoU gene was detected in 71.4% of the MDR P. aeruginosa isolates. These results indicate that the presence of the exoU gene can be a predictive marker for the persistence of MDR P. aeruginosa isolates.

Lung Adenocarcinoma Gene Mutation in Koreans: Detection Using Next Generation Sequence Analysis Technique and Analysis of Concordance with Existing Genetic Test Methods (한국인의 폐선암 유전자 돌연변이: 차세대 염기서열 분석법을 이용한 검출 및 기존 유전자 검사법과의 일치도 분석)

  • Jae Ha BAEK;Kyu Bong CHO
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.1
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    • pp.16-28
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    • 2023
  • Lung adenocarcinoma accounts for about 40% of all lung cancers. With the recent development of gene profiling technology, studies on mutations in oncogenes and tumor suppressor genes, which are important for the development and growth of tumors, have been actively conducted. Companion diagnosis using next-generation sequencing helps improve survival with targeted therapy. In this study, formalin-fixed paraffin-embedded tissues of non-small cell lung cancer patients were subjected to hematoxylin and eosin staining for detecting genetic mutations that induce lung adenocarcinoma in Koreans. Immunohistochemical staining was also performed to accurately classify lung adenocarcinoma tissues. Based on the results, next-generation sequencing was applied to analyze the types and patterns of genetic mutations, and the association with smoking was established as the most representative cause of lung cancer. Results of next-generation sequencing analysis confirmed the single nucleotide variations, copy number variations, and gene rearrangements. In order to validate the reliability of next-generation sequencing, we additionally performed the existing genetic testing methods (polymerase chain reaction-epidermal growth factor receptor, immunohistochemistry-anaplastic lymphoma kinase (D5F3), and fluorescence in situ hybridiation-receptor tyrosine kinase 1 tests) to confirm the concordance rates with the next-generation sequencing test results. This study demonstrates that next-generation sequencing of lung adenocarcinoma patients simultaneously identifies mutation.

Molecular Characterization of Vibrio parahaemolyticus Isolates from Seawater, Fish Tanks, and Distributed Fishery Products in Jeju (제주지역 해수, 수족관물, 유통수산물에서 분리된 장염비브리오균의 분자생물학적 특성)

  • Man Jae Cho;Eunok Kang;Ye-seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.246-254
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    • 2023
  • V. parahaemolyticus causes waterborne and foodborne disease such as acute diarrhea. In this study, V. parahaemolyticus isolates from seawater, fish tanks, and distributed fishery products in Jeju were investigated for potential toxin or species-specific genes (tdh, trh, tlh, and toxR) using RT-PCR and their genetic characteristics were analyzed using Pulsed-field gel electrophoresis (PFGE). Overall, V. parahaemolyticus of 90 strains (36.7%), including 33 strains from seawater, 8 strains from fish tanks, and 50 strains from fishery products, were isolated from 245 samples. All V. parahaemolyticus strains did not detect the tdh gene, whereas all strains detected tlh or toxR genes. In addition, trh genes were detected in 3 strains from seawater and 1 strain from fishery products. Monthly quantitative testing of seawater revealed that V. parahaemolyticus was positively correlated with water temperature. The 90 strains of V. parahemolyticus obtained in this study showed by gene homology between types, ranging from 64.0-97.3%. Among these, thirteen types showed 100% homology between genes. These results indicate that continuous monitoring is needed to facilitate food poisoning epidemiological investigations because some isolated V. parahaemolyticus strains harbored toxin genes and V. parahaemolyticus strains isolated from seawater, fish tanks, and distributed fishery products showed genetic similarity.

The gene expression programming method to generate an equation to estimate fracture toughness of reinforced concrete

  • Ahmadreza Khodayari;Danial Fakhri;Adil Hussein, Mohammed;Ibrahim Albaijan;Arsalan Mahmoodzadeh;Hawkar Hashim Ibrahim;Ahmed Babeker Elhag;Shima Rashidi
    • Steel and Composite Structures
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    • v.48 no.2
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    • pp.163-177
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    • 2023
  • Complex and intricate preparation techniques, the imperative for utmost precision and sensitivity in instrumentation, premature sample failure, and fragile specimens collectively contribute to the arduous task of measuring the fracture toughness of concrete in the laboratory. The objective of this research is to introduce and refine an equation based on the gene expression programming (GEP) method to calculate the fracture toughness of reinforced concrete, thereby minimizing the need for costly and time-consuming laboratory experiments. To accomplish this, various types of reinforced concrete, each incorporating distinct ratios of fibers and additives, were subjected to diverse loading angles relative to the initial crack (α) in order to ascertain the effective fracture toughness (Keff) of 660 samples utilizing the central straight notched Brazilian disc (CSNBD) test. Within the datasets, six pivotal input factors influencing the Keff of concrete, namely sample type (ST), diameter (D), thickness (t), length (L), force (F), and α, were taken into account. The ST and α parameters represent crucial inputs in the model presented in this study, marking the first instance that their influence has been examined via the CSNBD test. Of the 660 datasets, 460 were utilized for training purposes, while 100 each were allotted for testing and validation of the model. The GEP model was fine-tuned based on the training datasets, and its efficacy was evaluated using the separate test and validation datasets. In subsequent stages, the GEP model was optimized, yielding the most robust models. Ultimately, an equation was derived by averaging the most exemplary models, providing a means to predict the Keff parameter. This averaged equation exhibited exceptional proficiency in predicting the Keff of concrete. The significance of this work lies in the possibility of obtaining the Keff parameter without investing copious amounts of time and resources into the CSNBD test, simply by inputting the relevant parameters into the equation derived for diverse samples of reinforced concrete subject to varied loading angles.

A Familial Case with Holt-Oram Syndrome with a Novel TBX5 Mutation

  • Lee, Beom Hee;Kim, Yoo-Mi;Kim, Gu-Hwan;Kim, Young-Hwue;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.9 no.2
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    • pp.98-100
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    • 2012
  • Holt-Oram syndrome (HOS) is the most common heart-hand syndrome, which is inherited in an autosomal dominant manner, but most cases are sporadic. This condition is characterized by upper-extremity malformations involving radial-ray, thenar, and carpal bones, and congenital heart malformations including atrial septal defect and ventricular septal defect. It is caused by mutations in the TBX5 gene. In this report, a Korean case with HOS is described, which is inherited from her father. A novel nonsense mutation, $p.Glu294^*$, was identified. This is the first Korean case with HOS confirmed by genetic testing.