• Title/Summary/Keyword: PCR detection

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Chromosomal Localization and Mutation Detection of the Porcine APM1 Gene Encoding Adiponectin (Adiponectin을 암호화하는 돼지 APM1 유전자의 염색체상 위치파악과 돌연변이 탐색)

  • Park, E.W.;Kim, J.H.;Seo, B.Y.;Jung, K.C.;Yu, S.L.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.537-546
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    • 2004
  • Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.

Polymorphisms in Glutamate Receptor, Ionotropic, N-methyl-D-aspartate 2B(GRIN2B) Genes of Autism Spectrum Disorders in Korean Population : Family-based Association Study (한국인 자폐스펙트럼장애에서 Glutamate Receptor, Ionotropic, N-methyl-D-Aspartate 2B(GRIN2B) 유전자 다형성-가족기반연구)

  • Yoo, Hee Jeong;Cho, In Hee;Park, Mira;Yoo, Hanik K.;Kim, Jin Hee;Kim, Soon Ae
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.289-298
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    • 2006
  • Objectives : Autism is a complex neurodevelopmental spectrum disorder with a strong genetic component. Previous neurochemical and genetic studies suggested the possible involvement of glutamate N-methyl-D-aspartate(NMDA) receptor in autism. The aim of study was to investigate the association between the NMDA2B receptor gene(GRIN2B) and autism spectrum disorders(ASD) in the Korean population. Methods : The patients with ASD were diagnosed with Autism Diagnostic Interview-Revised and Autism Diagnostic Observation Schedule based on DSM-IV diagnostic classification. The present study was conducted with the detection of four single nucleotide polymorphisms(SNPs) in GRIK2 and family-based association analysis of the single nucleotide polymorphisms in Korean ASD trios using transmission disequilibrium test (TDT). Results : One hundred twenty six patients with ASD and their biological parents were analyzed. 86.5% were male and 85.1% were diagnosed as autistic disorder. The mean age was $71.9{\pm}31.6$ months(range : 26-185 months). We found that rs1805247 showed significantly preferential transmission(TDT ${\chi}^2$=12.8, p<0.001) in ASD. Conclusion : One SNP in GRIN2B gene was significantly associated with ASD in the Korean population. This result suggests the possible involvement of glutamate NMDA receptor gene in the development of ASD.

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Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

Potential Contamination Sources on Fresh Produce Associated with Food Safety

  • Choi, Jungmin;Lee, Sang In;Rackerby, Bryna;Moppert, Ian;McGorrin, Robert;Ha, Sang-Do;Park, Si Hong
    • Journal of Food Hygiene and Safety
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    • v.34 no.1
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    • pp.1-12
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    • 2019
  • The health benefits associated with consumption of fresh produce have been clearly demonstrated and encouraged by international nutrition and health authorities. However, since fresh produce is usually minimally processed, increased consumption of fresh fruits and vegetables has also led to a simultaneous escalation of foodborne illness cases. According to the report by the World Health Organization (WHO), 1 in 10 people suffer from foodborne diseases and 420,000 die every year globally. In comparison to other processed foods, fresh produce can be easily contaminated by various routes at different points in the supply chain from farm to fork. This review is focused on the identification and characterization of possible sources of foodborne illnesses from chemical, biological, and physical hazards and the applicable methodologies to detect potential contaminants. Agro-chemicals (pesticides, fungicides and herbicides), natural toxins (mycotoxins and plant toxins), and heavy metals (mercury and cadmium) are the main sources of chemical hazards, which can be detected by several methods including chromatography and nano-techniques based on nanostructured materials such as noble metal nanoparticles (NMPs), quantum dots (QDs) and magnetic nanoparticles or nanotube. However, the diversity of chemical structures complicates the establishment of one standard method to differentiate the variety of chemical compounds. In addition, fresh fruits and vegetables contain high nutrient contents and moisture, which promote the growth of unwanted microorganisms including bacterial pathogens (Salmonella, E. coli O157: H7, Shigella, Listeria monocytogenes, and Bacillus cereus) and non-bacterial pathogens (norovirus and parasites). In order to detect specific pathogens in fresh produce, methods based on molecular biology such as PCR and immunology are commonly used. Finally, physical hazards including contamination by glass, metal, and gravel in food can cause serious injuries to customers. In order to decrease physical hazards, vision systems such as X-ray inspection have been adopted to detect physical contaminants in food, while exceptional handling skills by food production employees are required to prevent additional contamination.

Epidemiology and Clinical Characteristics of Parainfluenza Virus Type 4 in Korean Children: a Single Center Study, 2015-2017 (소아에서 파라인플루엔자 바이러스 4형의 역학 및 임상 양상에 대한 단일기관 연구: 2015-2017)

  • Sohn, Young Joo;Choi, Youn Young;Yun, Ki Wook;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.156-164
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    • 2018
  • Purpose: We aimed to identify the epidemiology and the clinical characteristics of human parainfluenza virus type 4 (HPIV-4) infection compared to HPIVs 1-3 infections in Korean children. Methods: We reviewed medical records of children with HPIV infection who visited Seoul National University Children's Hospital from 2015 to 2017. Detection of respiratory viruses was performed using real time-polymerase chain reaction (rt-PCR), which could differentiate HPIVs 1-4. Diagnosis was classified as a febrile illness, upper respiratory tract infection (URI), croup, bronchiolitis, or pneumonia. The epidemiology, demographic features, and clinical characteristics among HPIV types were compared. The clinical data were analyzed only for the previously healthy children. Results: Of the 472 children diagnosed with HPIV infection, 108 (22.9%) were previously healthy: 24 (22.2%), 19 (17.6%), 39 (36.1%), and 26 (24.1%) in HPIV types 1, 2, 3, and 4, respectively. The median age of children with HPIV-4 infection was 11 (0-195) months: the proportion of children aged < 2 years and 2 to < 5 years were 65.4% and 19.2%, respectively. Clinical diagnoses of HPIV-4 infection were bronchiolitis (38.5%), pneumonia (30.8%), and URI (30.8%). Croup was the most prevalent in HPIV-2 (21.1%) and none in HPIV-4 infection (P=0.026). Hospital admission rates among HPIV types were not significantly different (P>0.05). Conclusions: We observed seasonal peak of HPIV-4 infection in 2015 and 2017. HPIV-4 was a common respiratory pathogen causing lower respiratory tract infection in hospitalized children.

Double-processed ginseng berry extracts enhance learning and memory in an Aβ42-induced Alzheimer's mouse model (Aβ42로 유도된 알츠하이머 마우스 모델에서 이중 가공 인삼열매 추출물의 학습 및 기억 손실 개선 효과)

  • Jang, Su Kil;Ahn, Jeong Won;Jo, Boram;Kim, Hyun Soo;Kim, Seo Jin;Sung, Eun Ah;Lee, Do Ik;Park, Hee Yong;Jin, Duk Hee;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.160-168
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    • 2019
  • This study aimed to determine whether double-processed ginseng berry extract (PGBC) could improve learning and memory in an $A\hat{a}42$-induced Alzheimer's mouse model. Passive avoidance test (PAT) and Morris water-maze test (MWMT) were performed after mice were treated with PGBC, followed by acetylcholine (ACh) measurement and glial fibrillary acidic protein (GFAP) detection for brain damage. Furthermore, acetylcholinesterase (AChE) activity and choline acetyltransferase (ChAT) expression were analyzed using Ellman's and qPCR assays, respectively. Results demonstrated that PGBC contained a high amount of ginsenosides (Re, Rd, and Rg3), which are responsible for the clearance of $A{\hat{a}} 42$. They also helped to significantly improve PAT and MWMT performance in the $A{\hat{a}} 42-induced$ Alzheimer's mouse model when compared to the normal group. Interestingly, ACh and ChAT were remarkably upregulated and AChE activities were significantly inhibited, suggesting PGBC to be a palliative adjuvant for treating Alzheimer's disease. Altogether, PGBC was found to play a positive role in improving cognitive abilities. Thus, it could be a new alternative solution for alleviating Alzheimer's disease symptoms.

Effect of Verapamil on Cellular Uptake of Tc-99m MIBI and Tetrofosmin on Several Cancer Cells (수종의 암세포에서 Verapamil이 Tc-99m MIBI와 Tetrofosmin의 섭취에 미치는 영향)

  • Kim, Dae-Hyun;Yoo, Jung-Ah;Suh, Myung-Rang;Bae, Jin-Ho;Jeong, Shin-Young;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.85-98
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    • 2004
  • Purpose: Cellular uptake of $^{99}mTc$-sestamibi (MIBI) and $^{99}mTc$-tetrofosmin (TF) is low in cancer cells expressing multidrug resistance(MDR) by p-glycoprotein(Pgp) or multidrug related protein(MRP). Verapamil is known to increase cellular uptake of MIBI in MDR cancer cells, but is recently reported to have different effects on tracer uptake in certain cancer cells. This study was prepared to evaluate effects of verapamil on cellular uptake of MIBI and TF in several cancer cells. Materials and Methods: Celluar uptakes of Tc-99m MIBI and TF were measured in erythroleukermia K562 cell, breast cancer MCF7 cell, and human ovarian cancer SK-OV-3 cells, and data were compared with those of doxorubicin-resistant K562(Ad) cells. RT-PCR and Western blot analysis were used for the detection of mdr1 mRNA and Pgp expression, and to observe changes in isotypes of PKC enzyme. Effects of verapamil on MIBI and TF uptake were evaluated at different concentrations upto $200{\mu}M\;at\;1{\times}10^6\;cells/ml\;at\;37^{\circ}C$. Radioactivity in supernatant and pellet was measured with gamma counter to calculate cellular uptake ratio. Toxicity of verapamil was measured with MTT assay. Results: Cellular uptakes of MIBI and TF were increased by time in four cancer cells studied. Co-incubation with verapamil resulted in an increase in uptake of MIBI and TF in K562(Adr) cell at a concentration of $100{\mu}M$ and the maximal increase at $50{\mu}M$ was 10-times to baseline. In contrast, uptakes of MIBI and TF in K562, MCF7, SK-OV3 cells were decreased with verapamil treatment at a concentration over $1{\mu}M$. With a concentration of $200{\mu}M$ verapamil, MIBI and TF uptakes un K562 cells were decreased to 1.5 % and 2.7% of those without verapamil, respectively. Cellular uptakes of MIBI and TF in MCF7 and SK-OV-3 cells were not changed with $10{\mu}M$, but were also decreased with verapamil higher than $10{\mu}M$, resulting 40% and 5% of baseline at $50{\mu}M$. MTT assay of four cells revealed that K562, MCF7, SK-OV3 were not damaged with verapamil at $200{\mu}M$. Conclusion: Although verapamil increases uptake of MIBI and TF in MDR cancer cells, cellular uptakes were further decreased with verapamil in certain cancer cells, which is not related to cytotoxicity of drug. These results suggest that cellular uptakes of both tracers might differ among different cells, and interpretation of changes in tracer uptake with verapamil in vitro should be different when different cell lines are used.