• Title/Summary/Keyword: reference gene

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Effectiveness of Krill Oil in Regulating Skin Moisture

  • Yoon-Seok Chun;Jongkyu Kim;Ji-Hoon Lim;Namju Lee;Sae-kwang Ku
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.359-368
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    • 2023
  • This study aims to explore the impact of Krill Oil (KO, SuperbaTM Boost) on skin moisturization regulation. The research involved five groups: an intact control, a reference group (L-AA 100 mg/kg), and KO groups (400, 200, and 100 mg/kg), each comprising ten mice. Oral administration was conducted for 8 weeks (56 days), during which changes in body weight, hyaluronan, collagen type 1 (COL1), transforming growth factor-β1 (TGF-β1), ceramide, and water contents were analyzed in dorsal back skin tissue. Real-time PCR was employed to assess gene expression related to hyaluronic acid synthesis (HAS1, HAS2, HAS3), COL1 synthesis (COL1A1 and COL1A2), and TGF-β1. Results demonstrated that KO administration significantly increased hyaluronan content, hyaluronic acid synthesis (HAS1, HAS2, HAS3), COL1 content, COL1 synthesis (COL1A1 and COL1A2), TGF-β1 content, TGF-β1 mRNA expression, ceramide content, and water content in a concentration-dependent manner compared to the intact control. Importantly, no discernible disparities were noted between the KO and L-AA groups, even though they received equivalent oral dosages. This study accentuates the potential utility of exogenous KO in the regulation of skin moisture, thus positioning it as a promising avenue for the development of nutricosmetics. Future research endeavors should delve into the role of KO in safeguarding against both intrinsic and extrinsic aging-related skin manifestations, as well as its potential to ameliorate skin wrinkles, in conjunction with its moisturizing attributes.

Current status and prospects of molecular marker development for systematic breeding program in citrus (감귤 분자육종을 위한 분자표지 개발 현황 및 전망)

  • Kim, Ho Bang;Kim, Jae Joon;Oh, Chang Jae;Yun, Su-Hyun;Song, Kwan Jeong
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.261-271
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    • 2016
  • Citrus is an economically important fruit crop widely growing worldwide. However, citrus production largely depends on natural hybrid selection and bud sport mutation. Unique botanical features including long juvenility, polyembryony, and QTL that controls major agronomic traits can hinder the development of superior variety by conventional breeding. Diverse factors including drastic changes of citrus production environment due to global warming and changes in market trends require systematic molecular breeding program for early selection of elite candidates with target traits, sustainable production of high quality fruits, cultivar diversification, and cost-effective breeding. Since the construction of the first genetic linkage map using isozymes, citrus scientists have constructed linkage maps using various DNA-based markers and developed molecular markers related to biotic and abiotic stresses, polyembryony, fruit coloration, seedlessness, male sterility, acidless, morphology, fruit quality, seed number, yield, early fruit setting traits, and QTL mapping on genetic maps. Genes closely related to CTV resistance and flesh color have been cloned. SSR markers for identifying zygotic and nucellar individuals will contribute to cost-effective breeding. The two high quality citrus reference genomes recently released are being efficiently used for genomics-based molecular breeding such as construction of reference linkage/physical maps and comparative genome mapping. In the near future, the development of DNA molecular markers tightly linked to various agronomic traits and the cloning of useful and/or variant genes will be accelerated through comparative genome analysis using citrus core collection and genome-wide approaches such as genotyping-by-sequencing and genome wide association study.

Noninvasive Prenatal Diagnosis using Cell-Free Fetal DNA in Maternal Plasma: Clinical Applications

  • Yang, Young-Ho;Han, Sung-Hee;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.1-16
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    • 2011
  • Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.

Promoter -202 A/C Polymorphism of Insulin-like Growth Factor Binding Protein-3 Gene and Non-small Cell Lung Cancer Risk (인슐린양 성장 인자 결합 단백-3 유전자 -202 좌위의 다형성에 따른 비소세포폐암의 위험도)

  • Moon, Jin Wook;Chang, Yoon Soo;Han, Chang Hoon;Kang, Shin Myung;Park, Moo Suk;Byun, Min Kwang;Chung, Wou Young;Park, Jae Jun;Yoo, Kyeong Nam;Shin, Ju Hye;Kim, Young Sam;Chang, Joon;Kim, Sung Kyu;Kim, Hee Jung;Kim, Se Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.359-366
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    • 2005
  • Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.

Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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A familial case of limb-girdle muscular dystrophy with CAV3 mutation

  • Lee, Seungbok;Jang, Sesong;Shim, Youngkyu;Kim, Woo Joong;Kim, Soo Yeon;Cho, Anna;Kim, Hunmin;Kim, Jong-Il;Lim, Byung Chan;Hwang, Hee;Choi, Jieun;Kim, Ki Joong;Chae, Jong Hee
    • Journal of Genetic Medicine
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    • v.16 no.2
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    • pp.67-70
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    • 2019
  • Limb-girdle muscular dystrophy (LGMD) is a group of muscular dystrophies that has extremely heterogeneous clinical features and genetic background. The caveolin-3 gene (CAV3) is one of the causative genes. LGMD appears as a clinical continuum, from isolated skeletal muscle involvement to long QT syndrome. Here we report two patients without apparent muscle weakness in a family with CAV3 mutation. A 7-month-old Korean boy visited our muscle clinic because of an incidental finding of elevated serum creatine kinase (CK) concentration (680 IU/L, reference range, 20-270 IU/L) without clinical symptoms. The patient was born after an uneventful pregnancy and showed normal developmental milestones. He developed pseudohypertrophy of his calf muscle during the follow-up. We obtained a muscle biopsy at age 14 months, which showed size variations and degenerating/regenerating myofibers with endomysial fibrosis and immunohistochemical evidence of normal dystrophin. Under the impression of LGMD, we performed target panel sequencing and identified a heterozygous in-frame mutation of CAV3, c.307_312delGTGGTG (p.Val103_Val104del). Immunohistochemical staining of muscle indicated complete loss of caveolin-3 compared with normal control muscle, which supported the variant's pathogenicity. We performed segregation analysis and found that the patient's mother had the same variant with elevated serum CK level (972 IU/L). We report on autosomal dominant familial caveolinopathy caused by a pathogenic variant in CAV3, which was asymptomatic until the fourth decade. This case highlights the utility of next generation sequencing in the diagnosis of muscular dystrophies and the additive role of muscle biopsy to confirm the variants.

Selective Isolation and Phylogeny of the Yeast Species Associated with Aloe vera and Aloe saponaria (알로에 베라(A. vera)와 알로에 사포나리아(A. saponaria)로 부터 효모의 분리 및 계통분석)

  • Choi, Sungchang;Kim, Myung-Uk;Kim, Jong-Shik
    • Korean Journal of Environmental Agriculture
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    • v.32 no.3
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    • pp.240-243
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    • 2013
  • BACKGROUND: Several yeast species have potential applications in biotechnology and the identification of such yeast species is of great interest. The first step in the identification of yeasts is the establishment of an effective isolation method. Thus, we compared the efficacy of different yeast media in the isolation of yeast associated with Aloe vera and Aloe saponaria. METHODS AND RESULTS: In this study, we spread homogenized A. vera and A. saponaria leaves onto 4 different yeast selective media containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose. We observed high selectivity for yeast and many colonies on media. We isolated 67 yeast strains from A. vera and 42 yeast strains from A. saponaria. We used phylogenetic analysis to identify the yeast isolates based on ITS region sequencing and performed sequence analysis on representative isolates from each agar plate. Further, we compared the sequences obtained with reference sequences. The yeast species isolated from A. vera were as follows: 56 isolates of Meyerozyma, 9 isolates of Cryptococcus, and 1 isolate each of Rhodotorula and Sporobolomyces. Those isolated from A. saponaria were as follows: 41 isolates of Rhodosporidium and 1 isolate of Sporobolomyces. CONCLUSION(S): All the isolates obtained using large agar plate containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose were identified as yeast. Therefore, we concluded that this method is useful for selective screening of yeast species.

Assessment of Genetic Diversity and Relationships Between Korean Cattle and Other Cattle Breeds by Microsatellite loci (Microsatellite loci 분석에 의한 한우와 타 품종간의 유전적 유연관계)

  • Yoon, D.H.;Park, E.W.;Lee, S.H.;Lee, H.K.;Oh, S.J.;Cheong, I.C.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.341-354
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    • 2005
  • For the genetic assessment of the cattle breeds including Hanwoo, eleven microsatellite markers on ten bovine autosomes were genetically characterized for 618 individuals of nineteen cattle breeds; North Eastern Asian breeds (Korean cattle, Korean Black cattle, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European Bas taurus (Angus, Hereford, Charolais, Holstein, Limousin), African Bas taurus (N'Dama, Baoule), African Bas indicus (Kavirondo Zebu, White Fulani), Asian Bas indicus (Sahiwal, Nelore) and one Bali cattle, Bas banteng as an outbreed-reference population. Allele frequencies derived from the genotyping data were used in estimating heterozygosities, gene diversities and genetic distances. The microsatellite loci were highly polymorphic, with a total of 162 different alleles observed across all loci. Variability in allele numbers and frequencies was observed among the breeds. The average expected heterozygosity of North Eastern Asian breeds was higher than those of European and African taurines, but lower than those of Asian and African indicines. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of the phylogenetic trees. The genetic distances between North Eastern Asian cattle breeds and Bas indicus were similar with those between European Bas taurus and Bas indicus, and African Bas taurus and Bas indicus, respectively. The clusters were clearly classified into North Eastern Asian, European and African taurines groups as well as different cluster with Chinese mainland breeds, firstly out-grouping with Bas indicus. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bas primigenius in Europe and Bas indicus in India and North Eastern Asian Bas taurus may be have separate domestication from European and African Bas taurus.

Detection of Red Sea Bream Iridovirus (RSIV) from marine fish in the Southern Coastal Area and East China Sea (남.서해안과 동중국해 자연산 어류에서 Red Sea Bream Iridovirus (RSIV)의 검출)

  • Lee, Wol-La;Kim, Seok-Ryel;Yun, Hyun-Mi;Kitamura, Shin Ichi;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.20 no.3
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    • pp.211-220
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    • 2007
  • Red sea bream iridovirus disease (RSIVD) cause massive economic losses in marine aquaculture industry in Korea. The causative agent of this disease (RSIV) infects a wide range of fish species. The aims of this study were to monitor RSIV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Polymerase Chain Reaction) PCR results showed that RSIV were detected in 39 (24.3%) out of 160 fish. MCP gene sequences of viral strains isolated in this study were closely related to that of a reference strain, red seabream-K, belonging to Megalocytivirus subgroup Ⅲ. The results suggest that some of wild marine fishes are RSIV carriers and may spread the pathogen directly to fish farmed in coastal area.

Studies on the safety of Brucella abortus RB51 vaccine I. Comparison of the biochemical and genetic characteristics of Brucella abortus RB51 vaccine strains (부루세라백신(RB51)의 안전성에 관한 연구 I. Brucella abortus RB51 백신균주의 생화학적 및 유전학적 성상비교)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Kang, Seung-won;Kim, Jong-yeom;Yoon, Yong-dhuk;Cho, Sang-nae;Yoo, Han-sang;Olsen, Steven C.
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.533-541
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    • 2000
  • Biochemical and genetic analysis were carried out to investigate the potential recovery of pathogenecity or related mutations of Brucella abortus RB51 vaccine strains. RB51 strains were recovered from commercial vaccines, including related seed stocks from private companies in Republic of Korea, strain from USA, a reference strain from C university and a field isolate (Daehungjin) from aborted dairy cow after RB51 vaccination were compared with two identified virulent wild strains (S2308 and a field strain isolated from dairy cow in Korea) at the same conditions. All the strains examined, except identified pathogenic strains, revealed the identical characteristics to the original RB51 in biochemical properties, antigen and bacteriophage typing. Outer membrane protein (OMP) profiles from strains of RB51 showed the same patterns with standard RB51 in SDS-PAGE. In addition, Western blotting with the brucella specific monoclonal antibody also indicated that all the vaccine strains were completely deficient in their LPS compared to the pathogenic Br abortus strains. The differences in DNA structures among strains were also possible to detect after PCR. All vaccine strains, except S19, S1119-3, S1075, S544 and Br suis, were amplified a 178bp DNA fragment of eri-gene, and 364bp of IS711 elements. In contrast, 498bp DNA product was only found with Br abortus. Overall evidences in the present study confirmed that the RB51 strains for vaccine production in Korea did not originated from the phenomena of possible recovery of pathogenicity or related to any potential mutation event at all.

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