• Journal of Veterinary Science
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• v.22 no.5
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• pp.63.1-63.14
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• 2021

Background: Recently, mesenchymal stem cells therapy has been performed in dogs, although the outcome is not always favorable. Objectives: To investigate the therapeutic efficacy of mesenchymal stem cells (MSCs) using dog leukocyte antigen (DLA) matching between the donor and recipient in vitro. Methods: Canine adipose-derived MSCs (cA-MSCs) isolated from the subcutaneous tissue of Dog 1 underwent characterization. For major DLA genotyping (DQA1, DQB1, and DRB1), peripheral blood mononuclear cells (PBMCs) from two dogs (Dogs 1 and 2) were analyzed by direct sequencing of polymerase chain reaction (PCR) products. The cA-MSCs were co-cultured at a 1:10 ratio with activated PBMCs (DLA matching or mismatching) for 3 days and analyzed for immunosuppressive (IDO, PTGS2, and PTGES), inflammatory (IL6 and IL10), and apoptotic genes (CASP8, BAX, TP53, and BCL2) by quantitative real-time reverse transcriptase-PCR. Results: cA-MSCs were expressed cell surface markers such as CD90⁺/44⁺/29⁺/45^- and differentiated into osteocytes, chondrocytes, and adipocytes in vitro. According to the Immuno Polymorphism Database, DLA genotyping comparisons of Dogs 1 and 2 revealed complete differences in genes DQA1, DQB1, and DRB1. In the co-culturing of cA-MSCs and PBMCs, DLA mismatch between the two cell types induced a significant increase in the expression of immunosuppressive (IDO/PTGS2) and apoptotic (CASP8/BAX) genes. Conclusions: The administration of cA-MSCs matching the recipient DLA type can alleviate the need to regulate excessive immunosuppressive responses associated with genes, such as IDO and PTGES. Furthermore, easy and reliable DLA genotyping technology is required because of the high degree of genetic polymorphisms of DQA1, DQB1, and DRB1 and the low readability of DLA 88.

• Korean Journal of Breeding Science
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• v.40 no.4
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• pp.387-393
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• 2008

Over 7 individual rice (Oryza sativa L.) plants per a line were sowed and sampled by pooled sampling method for genomic DNA extraction. The 5,400 flanking sequence tags (FSTs) were analysed by adaptor PCR and direct sequencing. FST analysis showed that the intragenic FSTs, the intergenic FSTs, and the original insertional sequences including hot spot covered 48.1% (2,597), 25.6% (1,383), and 25% (1,350), respectively. The 2,597 intragenic FSTs were used for genotyping to determine whether they are heterozygous or homozygous, and 1,393 core lines were selected. Among them, 422 knockout genes were distributed on chromosome 3, while 56 - 157 intragenic FSTs scattered on other chromosomes. Among 1,393 FSTs, known genes such as transcription factor covered 59.4% (827), while unknown genes such as expressed protein covered 40.6% (566). RT-PCR indicated that some core lines had no expression or decreased expression level in their knockout genes. It means that core lines are very useful knockout lines for functional genomic studies.

• Parasites, Hosts and Diseases
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• v.57 no.4
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• pp.341-357
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• 2019

Acanthamoeba, one of free-living amoebae (FLA), remains a high risk of direct contact with this protozoan parasite which is ubiquitous in nature and man-made environment. This pathogenic FLA can cause sight-threatening amoebic keratitis (AK) and fatal granulomatous amoebic encephalitis (GAE) though these cases may not commonly be reported in our clinical settings. Acanthamoeba has been detected from different environmental sources namely; soil, water, hotspring, swimming pool, air-conditioner, or contact lens storage cases. The identification of Acanthamoeba is based on morphological appearance and molecular techniques using PCR and DNA sequencing for clinico-epidemiological purposes. Recent treatments have long been ineffective against Acanthamoeba cyst, novel anti-Acanthamoeba agents have therefore been extensively investigated. There are efforts to utilize synthetic chemicals, lead compounds from medicinal plant extracts, and animal products to combat Acanthamoeba infection. Applied nanotechnology, an advanced technology, has shown to enhance the anti-Acanthamoeba activity in the encapsulated nanoparticles leading to new therapeutic options. This review attempts to provide an overview of the available data and studies on the occurrence of pathogenic Acanthamoeba among the Association of Southeast Asian Nations (ASEAN) members with the aim of identifying some potential contributing factors such as distribution, demographic profile of the patients, possible source of the parasite, mode of transmission and treatment. Further, this review attempts to provide future direction for prevention and control of the Acanthamoeba infection.

• The Korean journal of helicobacter and upper gastrointestinal research
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• v.18 no.4
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• pp.277-279
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• 2018

전 세계적으로 Helicobacter pylori의 항생제 내성률은 지속적으로 증가하고 있으며, 기존의 제균 치료에 실패한 H. pylori 감염 환자들에 대한 효과적인 구제요법(rescue therapy)의 필요성 역시 증가하고 있다. 이 연구는 두 개 기관, 공개, 평행 그룹, 무작위 배정 연구로서 불응성 H. pylori 감염 환자들의 구제요법으로 유전자형 내성을 기반한 치료(genotype resistance-guided therapy)와 경험적 치료(empirical therapy) 중 어느 것이 보다 효과적인지를 비교하고자 하였다. 2012년 10월부터 2017년 9월까지 20세 이상의 불응성 H. pylori 감염 환자들을 대상으로 하였으며, 불응성 H. pylori 감염은 과거 두 종류 이상의 H. pylori 제균 치료를 받았음에도 불구하고 H. pylori 제균에 실패한 환자들로 정의하였다. 이들에게서 한 군은 14일간의 유전자형 내성을 기반한 순차 치료(n=21 in trial 1, n=205 in trial 2)를, 다른 한 군은 환자들의 과거 제균 치료 종류를 감안한 14일간의 경험적 순차 치료(n=20 in trial 1, n=205 in trial 2)를 시행하였다. 순차 치료법은 첫 7일은 esomeprazole 40 mg과 amoxicillin 1 g을 하루 두 번 복용한 다음, 나머지 7일은 esomeprazole 40 mg과 metronidazole 500 mg, 그리고 1) levofloxacin 250 mg 또는 2) clarithromycin 500 mg 또는 3) tetracycline 500 mg을 하루 두 번 복용하는 것으로 구성하였다. 23S ribosomal RNA (rRNA)나 gyrase A에 대한 내성 관련 돌연변이 여부는 direct sequencing을 통한 중합효소연쇄반응(polymerase chain reaction, PCR) 검사를 이용하였고, 제균 성공 여부는 요소호기검사를 통해 확인하였다. 일차 결과 지표는 치료 방법에 따른 제균율로 정하였다. Trial 1에서는 tetracycline 대신 doxycycline 100 mg을 사용하였는데, 제균 성공률이 유전자형 내성을 기반한 치료군에서는 17명(81%), 경험적 치료군에서는 12명(60%)으로 나타났다(P=0.181). 하지만, 다른 순차 치료군들과 비교하였을 때, doxycycline을 포함한 순차 치료군의 제균율이 현저히 낮은 것으로 나타나서(15/26, 57.7%) doxycycline을 포함한 순차 치료법은 종결하기로 하고, trial 2부터는 doxycycline 대신 tetracycline으로 교체하여 연구를 지속하였다. Trial 2의 intention-to-treat (ITT) 분석 결과, 유전자형 내성을 기반한 치료군에서는 160/205명(78%), 경험적 치료군에서는 148/205명(72.2%)으로 두 그룹 간의 통계적인 제균율의 차이는 보여주지 못하였다(P=0.170). 부작용 및 환자 순응도에서도 양 군 간의 의미 있는 차이는 없었다. 따라서, 두 종류 이상 H. pylori 제균 치료에 실패한 환자들이라고 할지라도 기존의 제균 치료력을 바탕으로 적절한 경험적 치료를 시행하는 것은 유전자형 내성을 기반한 치료 정도의 효과는 있으며 접근성, 비용, 환자들의 선호도 등의 여러 가지 부가적인 사항들을 고려할 때, 제균 치료력을 고려한 경험적 치료는 간단한 수준의 유전자형 내성을 기반한 치료의 대안으로 받아들여질 수 있을 것으로 제안하였다.

• Research in Plant Disease
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• v.26 no.4
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• pp.283-288
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• 2020

In September 2017, vein clearing and yellowing symptoms resembling those caused by viruses were observed on leaves of Malva verticillata in Chungnam, Korea. Nucleic acids were extracted from leaves of five symptomatic plants and tested by reverse transcription polymerase chain reaction using four virus specific primer pairs including malva vein clearing virus (MVCV). Amplicons of the expected size (600 bp) were obtained from total RNA of all samples using the MVCV-specific primers. To confirm the presence of MVCV in symptomatic plants, the DNA fragments from three samples were purified, and directly sequenced. BLAST analysis revealed that it shared the highest nucleotide identity (99%) with a MVCV isolate from tomato (Mexico). The virus isolates obtained from the third re-inoculated Chenopodium was designated as Cm1-5. Tissue from Cm1, Cm3, and Cm5 isolates was mechanically sap inoculated into 23 indicator plants. Cm3 isolate induced chlorotic local and mosaic symptoms in Althaea rosea. Phylogenetic analysis based on coat protein gene of 19 MVCV isolates from 6 different countries and plant species, did not correlated with either the geographical origin of the isolates, or pathogenicity. To our knowledge, this study first reports the natural occurrence of MVCV on M. verticillata in Korea and characterization of three Korean isolates of MVCV.

• Korean Journal of Food Science and Technology
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• v.53 no.1
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• pp.1-11
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• 2021

In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.

• Genomics & Informatics
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• v.20 no.3
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• pp.29.1-29.12
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• 2022

Several studies have shown associations between irinotecan toxicity and UGT1A genetic variations in colorectal and lung cancer, but only limited data are available for gastric cancer patients. We evaluated the frequencies of UGT1A polymorphisms and their relationship with clinicopathologic parameters in 382 Korean gastric cancer patients. Polymorphisms of UGT1A1*6, UGT1A1*27, UGT1A1*28, UGT1A1*60, UGT1A7*2, UGT1A7*3, and UGT1A9*22 were genotyped by direct sequencing. In 98 patients treated with irinotecan-containing regimens, toxicity and response were compared according to the genotype. The UGT1A1*6 and UGT1A9*22 genotypes showed a higher prevalence in Korean gastric cancer patients, while the prevalence of the UG1A1*28 polymorphism was lower than in normal Koreans, as has been found in other studies of Asian populations. The incidence of severe diarrhea after irinotecan-containing treatment was more common in patients with the UGT1A1*6, UGT1A7*3 and UGT1A9*22 polymorphisms than in controls. The presence of the UGT1A1*6 allele also showed a significant association with grade III-IV neutropenia. Upon haplotype and diplotype analyses, almost every patient bearing the UGT1A1*6 or UGT1A7*3 variant also had the UGT1A9*22 polymorphism, and all severe manifestations of UGT1A polymorphism-associated toxicity were related to the UGT1A9*22 polymorphism. By genotyping UGT1A9*22 polymorphisms, we could identify high-risk gastric cancer patients receiving irinotecan-containing chemotherapy, who would experience severe toxicity. When treating high-risk patients with the UGT1A9*22 polymorphism, clinicians should closely monitor them for signs of toxicity such as severe diarrhea or neutropenia.

• Journal of Life Science
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• v.33 no.8
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• pp.605-611
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• 2023

Recently, sporadic cases of human infection by genetic reassortants of H7Nx influenza A viruses have been reported; such viruses have also been continuously isolated from avian species. In this study, A/wild bird/South Korea/sw-anu/2023, a novel reassortant of the H7N1 avian influenza virus, was analyzed using full-genome sequencing and molecular characterization. Phylogenetic analysis showed that A/wild bird/South Korea/sw-anu/2023 belonged to the Eurasian lineage of H7Nx viruses. The polymerase basic (PB)2, PB1, polymerase acidic (PA), and nucleoprotein (NP) genes of these viruses were found to be closely related to those of avian influenza viruses isolated from wild birds, while the hemagglutinin (HA), neuraminidase (NA), matrix (M), and nonstructural (NS) genes were similar to those of avian influenza viruses isolated from domestic ducks. In addition, A/wild bird/South Korea/sw-anu/2023 also had a high binding preference for avian-specific glycans in the solid-phase direct binding assay. These results suggest the presence of a new generation of H7N1 avian influenza viruses in wild birds and highlight the reassortment of avian influenza viruses found along the East Asian-Australasian flyway. Overall, H7Nx viruses circulate worldwide, and mutated H7N1 avian viruses may infect humans, which emphasizes the requirement for continued surveillance of the H7N1 avian influenza virus in wild birds and poultry.

• Journal of Marine Life Science
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• v.8 no.2
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• pp.128-135
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• 2023

The present study was tried to identify whether the eel's larva was close to a conger (Conger myriaster), a pipe conger (Muraenesox cinereus) or four species of Anguilla. Experimental fishes were collected by set net in the gulf of enggang, Namhae, Korea from May to June. Their morphological characteristics were compared with adult fishes of a conger, a pipe conger and four species of Anguilla. For genetic classification, DNA was isolated and amplified by using 12S rRNA and 16S rRNA primer set. The PCR products were direct sequencing in both directions. The nucleotide sequences were analyzed using softwares. As results of morphological measurement on eel's larva, the percentages of head length and preanal length against total length were similar with a conger. Based on the nucleotide sequences, the phylogenetic tree also revealed a close relationship to a conger. Therefore, eel's larva, caught in Namhae from May to June, was identified into a conger's larva.

• Clinical and Experimental Pediatrics
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• v.51 no.2
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• pp.150-155
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• 2008

Purpose : It has been known that breast milk cause prolonged unconjugated hyperbilirubinemia. UGT1A1 is a important gene of uridine diphosphate glucuronosyltransferase (UGT) which has a major role of bilirubin metabolism. These findings suggest that there is a relationship between UGT1A1 gene mutation and prolonged jaundice of breast feeding infant. The aim of study was to investigate whether a polymorphism of the UGT1A1 gene exist in prolonged hyperbilirubinemia of breast milk feeding Korean infant. Methods : The genomic DNA was isolated from 50 full term Korean neonates, who had greater than a 10 mg/dL of serem bilirubin after 2 weeks of birth with no significant cause, and the other genomic DNA was isolated from 162 full term Korean neonates of the control population. Both group fed breast milk. We performed direct sequencing of TATA box and Gly71Arg polymorphism of the UGT1A1 gene. Results : Two of the 50 neonates with hyperbilirubinemia had AA polymorphism, and 40 had GA polymorphism. Five of the 129 neonates of the control group had AA polymorphism, and 4 had GA polymorphism. The allele frequency of G>A polymorphism in the hyperbilirubinemia group was 44.0%; it was significantly higher than 5.4% of the control group. TATA box polymorpism was not different both group significantly. Conclusion : Our result indicated that Gly71Arg polymorphism is associated with the prolonged hyperbilirubinemia of breast milk-feeding infant in Korean, while TATA box polymorphism is not associated with the prolonged hyperbilirubinemia of breast milk-feeding infant in Korean.