• Title/Summary/Keyword: Gene testing

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Evaluation of Loop Mediated Isothermal Amplification Based Methods for the Detection of African Swine Fever Virus from Food Waste

  • Siwon Lee;Junhwa Kwon;Su Hyang Kim;Jin-Ho Kim;Jaewon Jung;Kyung-Jin Lee;Ji-Yeon Park;Taek-Kyun Choi;Jun-Gu Kang;Tae Uk Han
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.334-339
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    • 2022
  • African swine fever virus (ASFV) is a highly contagious and lethal pathogen that poses a threat to the global pork industry. The World Organization for Animal Health (WOAH) has placed strict surveillance measures for ASFV. The possibility of long-term survival of ASFV in raw meat or undercooked pork has been reported. Accordingly, the problem of secondary infection in food waste from households or waste disposal facilities has emerged, raising the need for ASFV monitoring of food waste. However, most of the previously reported ASFV gene detection methods are focused on clinical monitoring of pigs. There are very few cases in which their application in waste has been verified. Since ASFV diagnosis requires rapid monitoring and immediate action, loop-mediated isothermal amplification (LAMP) may be suitable, but this requires conformity assessment for LAMP to be used as a diagnostic technique. In this study, six LAMP methods were evaluated, and two methods (kit and manual) were recommended for use in diagnosing ASFV in food waste.

Anti-oxidant Activity and Skin Barrier Function Measured in Different Solvent Fractions Obtained from Agrimoniae Herba (용아초(龍芽草) 유기용매 분획물의 항산화 및 피부 장벽 개선 효과)

  • Jegeun Han;Min Ju Kim;Hae-Jin Park;Kyung-Ha Lee;Seong-Soo Roh
    • The Korea Journal of Herbology
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    • v.38 no.3
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    • pp.19-26
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    • 2023
  • Objectives : Agrimoniae Herba is a herbal medicine widely distributed in Asia and contains flavonoids including catechin, quercitrin, rutin, hyperoside, and quercetin. This study aimed to investigate the anti-oxidant activity and skin barrier function of different solvent fractions (Hexane; methylene chloride, MC; ethyl acetate, EA; n-butanol, Bu; Water) obtained from Agrimoniae Herba. Methods : Anti-oxidant activity of different solvent fractions obtained from Agrimoniae Herba was investigated through total polyphenol contents, total flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity measurements. Then, filament aggregating protein (Filaggrin), Type I collagen, ceramide synthase (CERS) 3, and CERS4 were analyzed to evaluate the skin barrier strengthening effect of different solvent fractions obtained from Agrimoniae Herba on UVB-stimulated HaCaT cells. Results : As a result of measuring total polyphenol contents, total flavonoid contents, DPPH free radical scavenging activity, and ABTS radical scavenging activity, antioxidant activity was found to be excellent in the order of EA > Bu > MC > Hexane > Water. As a result of measuring mRNA gene expression of Type I collagen, Filaggrin, CERS3, and CERS4 after UVB-stimulated was applied to HaCaT cells treated with different solvent fractions obtained from Agrimoniae Herba, it was found to increase significantly in the Bu-treated group. Conclusion : Our findings show that the Bu sample obtained from Agrimoniae Herba has excellent anti-oxidant ability, which increases Type I collagen, Filaggrin, and ceramide synthetase in UVB-stimulated HaCaT cells to control the skin barrier improvement effect.

Assessment of lactic acid bacteria isolated from the chicken digestive tract for potential use as poultry probiotics

  • Merisa Sirisopapong;Takeshi Shimosato;Supattra Okrathok;Sutisa Khempaka
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1209-1220
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    • 2023
  • Objective: The use of probiotics as an alternative to antibiotics in animal feed has received considerable attention in recent decades. Lactic acid bacteria (LAB) have remarkable functional properties promoting host health and are major microorganisms for probiotic purposes. The aim of this study was to characterize LAB strains of the chicken digestive tract and to determine their functional properties for further use as potential probiotics in poultry. Methods: A total of 2,000 colonies were isolated from the ileum and cecal contents of the chickens based on their phenotypic profiles and followed by a preliminary detection for acid and bile tolerance. The selected 200 LAB isolates with exhibited well-tolerance in acid and bile conditions were then identified by sequencing the 16S rDNA gene, followed by acid and bile tolerance, antimicrobial activity, adhesion to epithelial cells and additional characteristics on the removal of cholesterol. Then, the two probiotic strains (L. ingluviei and L. salivarious) which showed the greatest advantage in vitro testing were selected to assess their efficacy in broiler chickens. Results: It was found that 200 LAB isolates that complied with all measurement criteria belonged to five strains, including L. acidophilus (63 colonies), L. ingluviei (2 colonies), L. reuteri (58 colonies), L. salivarius (72 colonies), and L. saerimneri (5 colonies). We found that the L. ingluviei and L. salivarius can increase the population of LAB and Bifidobacterium spp. while reducing Enterobacteria spp. and Escherichia coli in the cecal content of chickens. Additionally, increased concentrations of valeric acid and short chain fatty acids were also observed. Conclusion: This study indicates that all five Lactobacillus strains isolated from gut contents of chickens are safe and possess probiotic properties, especially L. ingluviei and L. salivarius. Future studies should evaluate the potential for growth improvement in broilers.

A Novel SLC25A15 Mmutation Causing Hyperornithinemia-Hyperammonemia-Homocitrullinuria Syndrome (Hyperornithinemia-hyperammonemia-homocitrullinuria 증후군을 유발하는 SLC25A15 유전자의 새로운 변이)

  • Jang, Kyung Mi;Hyun, Myung Chul;Hwang, Su-Kyeong
    • Journal of the Korean Child Neurology Society
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    • v.25 no.3
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    • pp.204-207
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    • 2017
  • Hyperornithinemia-hyperammonemia-homocitrullinuria syndrome (HHH syndrome) is a neurometabolic disorder with highly variable clinical severity ranging from mild learning disability to severe encephalopathy. Diagnosis of HHH syndrome can easily be delayed or misdiagnosed due to insidious symptoms and incomplete biochemical findings, in that case, genetic testing should be considered to confirm the diagnosis. HHH syndrome is caused by biallelic mutations of SLC25A15, which is involved in the urea cycle and the ornithine transport into mitochondria. Here we report a boy with spastic paraplegia and asymptomatic younger sister who have compound heterozygous mutations of c.535C>T (p.R179*) and c.116C>A (p.T39K) in the SLC25A15 gene. We identified that p.T39K mutation is a novel pathogenic mutation causing HHH syndrome and that p.R179*, which is prevalent in Japanese and Middle Eastern heritage, is also found in the Korean population.

Imaging Techniques and Differential Diagnosis for Inflammatory Bowel Disease (염증성 장질환의 영상기법 및 감별진단)

  • Kyoung Doo Song
    • Journal of the Korean Society of Radiology
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    • v.84 no.3
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    • pp.536-549
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    • 2023
  • The two main types of inflammatory bowel disease (IBD) are Crohn's disease and ulcerative colitis. Currently, when IBD is suspected, CT enterography is widely used as an initial imaging test because it can evaluate both the bowel wall and the outside of the bowel, helping to differentiate IBD from other diseases. When IBD is suspected, it is necessary to distinguish between Crohn's disease and ulcerative colitis. In most cases this is not difficult; however, in some cases, it is difficult and such cases are called IBD-unclassified. CT findings are often non-specific for ulcerative colitis, making it difficult to differentiate it from other diseases using imaging alone. In contrast, characteristic CT findings for Crohn's disease are often helpful in diagnosis, although diseases, such as tuberculous enteritis can mimic Crohn's disease. Recently, mutations in the gene encoding a prostaglandin transporter called SLCO2A1 have been discovered as the cause of the disease in some patients with multiple ulcers and strictures, similar to Crohn's disease. Therefore, genetic testing is being used to make a differential diagnosis.

Predicting tensile strength of reinforced concrete composited with geopolymer using several machine learning algorithms

  • Ibrahim Albaijan;Hanan Samadi;Arsalan Mahmoodzadeh;Danial Fakhri;Mehdi Hosseinzadeh;Nejib Ghazouani;Khaled Mohamed Elhadi
    • Steel and Composite Structures
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    • v.52 no.3
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    • pp.293-312
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    • 2024
  • Researchers are actively investigating the potential for utilizing alternative materials in construction to tackle the environmental and economic challenges linked to traditional concrete-based materials. Nevertheless, conventional laboratory methods for testing the mechanical properties of concrete are both costly and time-consuming. The limitations of traditional models in predicting the tensile strength of concrete composited with geopolymer have created a demand for more advanced models. Fortunately, the increasing availability of data has facilitated the use of machine learning methods, which offer powerful and cost-effective models. This paper aims to explore the potential of several machine learning methods in predicting the tensile strength of geopolymer concrete under different curing conditions. The study utilizes a dataset of 221 tensile strength test results for geopolymer concrete with varying mix ratios and curing conditions. The effectiveness of the machine learning models is evaluated using additional unseen datasets. Based on the values of loss functions and evaluation metrics, the results indicate that most models have the potential to estimate the tensile strength of geopolymer concrete satisfactorily. However, the Takagi Sugeno fuzzy model (TSF) and gene expression programming (GEP) models demonstrate the highest robustness. Both the laboratory tests and machine learning outcomes indicate that geopolymer concrete composed of 50% fly ash and 40% ground granulated blast slag, mixed with 10 mol of NaOH, and cured in an oven at 190°F for 28 days has superior tensile strength.

Effect of NaCl on the Stability of Oncolytic Vaccinia Virus (항암 백시니아 바이러스의 안전성에 대한 염화나트륨의 효과)

  • Kim, Seong-Geun;Ran, Gui Shao;Kwon, Hyuk-Chan;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.26 no.1
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    • pp.23-33
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    • 2016
  • Pexa-Vec (JX-594) is a specific cancer-targeted oncolytic and immunotherapeutic vaccinia virus. The purpose of this study was to develop methods to maximize the stability of Pexa-Vec. In short-term instability testing, viral activity was rapidly decreased both at 4℃ and at room temperature (RT), but it was completely restored after sonication followed by vortex. Long-term stability testing of Pexa-Vec in the following liquid formulations was performed: (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6, (C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, and (E) 30 mM Tris/pH 8.6, 15% sucrose. Viral activity decreased less than 2 log10 at 4℃, and RT was observed in 3 days in B, while viral activity was not decreased even after 4–8 weeks at 4℃ and at 1 week in RT in A, suggesting that neutral pH may be essential to maintain virus stability. The addition of 15% sucrose into A (D) significantly increased viral stability at −20℃, 4℃, or RT, and it was also observed at pH 8.6 (E). The addition of 150 mM NaCl into D (C) significantly increased viral stability in addition to the sucrose effect at 4℃ or RT. Accordingly, the viral activity in formulation C was maintained for 1.5 years at 4℃, and for 1-2 weeks in RT. In conclusion, we propose that formulation C can provide the most adequate condition for the proper storage of vaccinia oncolytic virus.

Angiotensin-converting Enzyme Gene Polymorphism and Cerebrovascular Disease in Korean population (한국인의 ACE(Angiotensin-converting Enzyme) 유전자의 다형성과 뇌혈관 질환과의 관계에 대한 연구)

  • Lee Jin Woo;Lee Kyung Jin;Rho Sam Woong;Kim Jae Jong;Bae Hyung Sup;Hong Moo Chang;Shin Min Kyu;Kim Young Suk;Bae Hyun Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.724-728
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    • 2002
  • Angiotensin-converting enzyme (ACE) gene polymorphism, which consists of presence (insertion, I) or absence (deletion, D) of a 250-bp fragment, is associated with ischemic heart disease, renovascular disease, systemic lupus erythematosus. Subjects with the DD genotype have higher levels of circulating ACE than subjects with the II genotype and show an increased tendency towards vascular wall thickness and contribute to the development of vascular disease. But the association between I/D polymorphism of the ACE gene and cerebrovascular disease is still controversial. The aim of this study was to determine whether the DNA polymorphism of the ACE are associated with cerebrovascular disease in Korean population. The study group comprised 377 Korean patients admitted to Kyunghee Oriental Medical Center in the year of 2000 for the treatment of brain infarction or brain hemorrhage. Magnetic resonance imaging(MRI) was performed for each patient to determine the stroke phenotype, infarction or hemorrhage. The 183 subjects without evidence of brain infarction or brain hemorrhage were selected from the some ethnical population(control group). Venous blood samples were drawn from each subject for the extraction of DNA. Genotypes of ACE were determined by polymerase chain reaction amplification of the genomic DNA. Case and control genotype frequencies were compared by chi-square testing. Both the patients and the controls were classified respectively into 4 groups: age less than forty years, age forty one to fifty, age fifty one to sixty, age greater than sixty years. There were no significant differences in the distributions of ACE genotypes among the patients with infarction, with hemorrhage and controls (Infarction: D/D 15.8%, I/D 46.7%, I/I 37.5%, Hemorrhage: D/D 15.1%, I/D 46.5%, I/I 38.4%, Control: D/D 18.6%, I/D 50.3%, I/I 31.2%). There was a significant difference in the distribution of ACE genotypes between the age greater than sixty year subgroup of patient with brain hemorrhage and the control (Hemorrhage: D/D 0%, I/D 55.6%, I/I 44.4%, Control: D/D 13.0%, I/D 63.0%, I/I 23.9%; Pearson Chi-Square value 5.956, P<0.05). Furthermore, the frequency of the ACE D/D type declined with increasing age both in the patient and control group (Patient group: age < 50 D/D 21.5%, age > 50 D/D 14.42%; Control group: age < 50 D/D 21.0%, age > 50 D/D 14.2%). In conclusion there is no clear association between ACE polymorphism and cerebrovascular disease in Korean population. Although, there was a tendency for the frequency of the ACE D/D type declined with increasing age in both patients and controls.

Diagnostic testing for Duchenne/Becker Muscular dystrophy using Dual Priming Oligonucleotide (DPO) system (Dual Priming Oligonucleotide (DPO) system을 이용한 듀시엔/베커형 근이영양증 진단법)

  • Kim, Joo-Hyun;Kim, Gu-Hwan;Lee, Jin-Joo;Lee, Dae-Hoon;Kim, Jong-Kee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.15-20
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    • 2008
  • Purpose : Large exon deletions in the DMD gene are found in about 60% of DMD/BMD patients. Multiplex PCR has been employed to detect the deletion mutation, which frequently generates noise PCR products due to the presence of multiple primers in a single reaction as well as the stringency of PCR conditions. This often leads to a false-negative or false-positive result. To address this problematic issue, we introduced the dual primer oligonucleotide (DPO) system. DPO contains two separate priming regions joined by a polydeoxyinosine linker that results in high PCR specificity even under suboptimal PCR conditions. Methods : We tested 50 healthy male controls, 50 patients with deletion mutation as deletion-positive patient controls, and 20 patients with no deletions as deletion-negative patient controls using DPO-multiplex PCR. Both the presence and extent of deletion were verified by simplex PCR spanning the promoter region (PM) and 18 exons including exons 3, 4, 6, 8, 12, 13, 17, 19, 43-48, 50-52, and 60 in all 120 controls. Results : DPO-multiplex PCR showed 100% sensitivity and specificity for the detection a deletion. However, it showed 97.1% sensitivity and 100% specificity for determining the extent of deletions. Conclusion : The DPO-multiplex PCR method is a useful molecular test to detect large deletions of DMD for the diagnosis of patients with DMD/BMD because it is easy to perform, fast, and cost-effective and has excellent sensitivity and specificity.

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Population-Based Newborn Hearing Impairment Screening Test Using GJB2 Mutation Analysis

  • Lee, Kyung-Ok;Jeong, Su-Jin;Byun, Ji-Young;Kim, Jeong-Sook;Lee, Hye-Jung;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.113-121
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    • 2007
  • Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

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