• Title/Summary/Keyword: late gene

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Expression Analysis of Lily Type Lectin Isotypes in the Rock Bream, Oplegnathus fasciatus: in the Tissue, Developmental Stage and Viral Infection

  • Lee, Young Mee;Yang, In Jung;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Noh, Gyeong Eon;Kim, Woo-Jin;Kim, Kyung-Kil
    • Development and Reproduction
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    • v.20 no.4
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    • pp.297-304
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    • 2016
  • Lectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

Effect of Leptin on the Expression of Lipopolysaccharide-Induced Chemokine KC mRNA in the Mouse Peritoneal Macrophages

  • Lee, Dong-Eun;Kim, Hyo-Young;Song, In-Hwan;Kim, Sung-Kwang;Seul, Jung-Hyun;Kim, Hee-Sun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.722-729
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    • 2004
  • Leptin is an adipocyte-secreted hormone and its plasma levels correlate with total body fat mass, however, it also plays a regulatory role in immunity, inflammation, and hematopoiesis. Chemokine is known as a chemoattractant cytokine in inflammatory reaction, but its role in leptin reaction has not been well studied. In this study, the direct effect of leptin on the expression of chemokine mRNAs and lipopolysaccharide (LPS)-induced chemokine KC mRNA in mouse peritoneal macrophages was investigated. Leptin did not induce the expression of lymphotactin, RANTES, eotaxin, MIP-1$\beta$, MIP-1$\alpha$, MIP-2, MCP-1, IP-10, TCA-3, and KC mRNA in mouse peritoneal macrophages, and had no direct effect on the expression of these LPS-induced chemokine mRNAs except KC mRNA. The synergistic effect of leptin on the expression of LPS-induced KC mRNA occurred late in the time course of response to LPS. The increased expressions of Ob-Rb mRNA and leptin receptor protein were detected during the LPS treatment. Leptin produced a substantial increase in the stability of the LPS-induced KC mRNA, and the synergistic effect of leptin on LPS-induced KC mRNA expression was further augmented by cycloheximide (CHX). Pyrrolidine dithiocarbamate (PDTC) did not block the synergistic effect of leptin on LPS-induced KC mRNA expression in mouse peritoneal macrophages. These data suggest that although leptin has no direct effect on the expression of lymphotactin, RANTES, eotaxin, MIP-1$\beta$, MIP-1$\alpha$, MIP-2, MCP-1, IP-10, TCA-3, and KC mRNA in mouse peritoneal macrophages, the synergistic effect of leptin on the expression of LPS-induced KC mRNA has the possibility that LPS might induce the expression of the Ob-Rb receptor or an unknown gene(s) that sensitizes macrophages to the synergistic function of leptin. Therefore, further studies are necessary to examine leptin as a regulatory factor of chemokine production.

Breeding of a New Japanese Apricot (Prunus mume Siebold et Zucc.) Cultivar 'Okjoo' with High Yields (다수성 매실 품종 '옥주')

  • Kim, Yoon-Kyeong;Kang, Sam-Seok;Choi, Jang-Jeon;Cho, Kwang-Sik;Won, Kyeong-Ho;Lee, Han-Chan;Choi, Jin-Ho
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.912-916
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    • 2014
  • Japanese apricot (Prunus mume Siebold and Zucc.) is a deciduous tree of the family Rosaceae, and it has long been used as a folk remedy for cough and dyspepsia. A new cultivar 'Okjoo' was developed from a cross between 'Gyokuei' and 'Rinsyu' carried out at the National Institute of Horticultural & Herbal Science in 1993. It w as s elected for good shape, large size and high yield capacity in 2006, and then it was granted official patent No. 4556 in 2013. It blooms 4 days and 2 days earlier than 'Gyokuei' and 'Rinsyu', respectively. Its flower petal color is pink, and the pollen amount is negligible. Its S-genotype, determined using Polymerase Chain Reaction with a S-RNase gene-specific primer pair, is $S_3S_6$. The average optimum harvest time of 'Okjoo' is late June. The fruit is round in shape and its suture is shallow. Average fruit weight is 18.5 g, and it contains total soluble solids $7.66^{\circ}Brix$ and titratable acidity at 4.81%. Fruit skin color is green. Sometimes only the light side of the fruits seems to develop blush. The incidence levels of scab (Cladosporium carpophilum Thumen) and bacterial shot hole (Xanthomonas arboricola pv. Pruni) are quite low. Consequently, 'Okjoo' seems to be a promising new cultivar for Japanese apricot growers.

Effect of Safflower and Safflower Seed Extract on Osteogenic Differentiation of MC3T3E1 Cells (홍화, 홍화씨 추출물이 MC3T3E1 세포의 골분화 과정에 미치는 영향)

  • Yu, Sung-ryul;Shin, Seon-mi
    • The Journal of Internal Korean Medicine
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    • v.36 no.4
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    • pp.518-526
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    • 2015
  • Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.

Effects of Estradiol-$17{\beta}$ and Nonylphenol on mRNA Expression of Estrogen Receptor-related Receptor $\beta$ Like 1 and Early Embryogensis in Sea Urchin, Strongylocentrotus nudus (Estradiol-$17{\beta}$와 Nonylphenol이 둥근성게(Strongylocentrotus nudus) 초기 배발생과 Estrogen Receptor-related Receptor $\beta$ Like 1 mRNA 발현에 미치는 영향)

  • Jung, Yu-Jung;Maeng, Se-Joeng;Sohn, Young-Chang
    • Development and Reproduction
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    • v.11 no.3
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    • pp.179-185
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    • 2007
  • The estrogens and estrogenic endocrine disrupting chemicals(EDCs) function through a steroid nuclear receptor-mediated process and subsequently regulate the transcription of mRNA for a number of target proteins. The estrogen receptor-related receptors(ERRs), which are structurally similar to estrogen receptors, are members of orphan nuclear receptor in the nuclear receptor superfamily and their functions are known to be involved in the formation of extra-embryonic ectoderm. To investigate effects of EDCs on early embryogenesis and ERR gene expression in marine invertebrates, we examined morphological changes and the mRNA expression of $ERR{\beta}$ like 1 in sea urchin Strongylocentrotus nudus exposed to estradiol-$17{\beta}(E_2)$ or nonylphenol(NP). The $E_2$ and NP-exposed embryos showed a delayed development compared to control embryos. Furthermore, they showed abnormal embryonic developments at late stages, i.e., blastular, gastrula and plutei stages. The mRNA level of $ERR{\beta}$ like 1 at the gastrula stage was significantly lower in $E_2$ and NP-exposed embryos than those of control group. These results suggest that NP and $E_2$ are potent chemicals causing abnormal embryonic development of S. nudus through at least in part down-regulated $ERR{\beta}$ like 1.

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Clinical Features, Response to Treatment, Prognosis, and Molecular Characterization in Korean Patients with Inherited Urea Cycle Defects

  • Yoo, Han-Wook;Kim, Gu-Hwan;Seo, Eul-Ju
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.2 no.1
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    • pp.77-79
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    • 2002
  • The urea cycle, consisting of a series of six enzymatic reactions, plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Five well characterized diseases have been described, resulting from an enzymatic defect in the biosynthesis of one of the normally expressed enzyme. This presentation will focus on two representative diseases; ornithine transcarbamylase(OTC) deficiency and citrullinemia(argininosuccinate synthetase deficiency). OTC deficiency is one of the most common inborn error of urea cycle, which is inherited in X-linked manner. We identified 17 different mutations in 20 unrelated Korean patients with OTC deficiency; L9X, R26P, R26X, T44I, R92X, G100R, R141Q, G195R, M205T, H214Y, D249G, R277W, F281S, 853 del C, R320X, V323M and 10 bp del at nt. 796-805. These mutations occur at well conserved nucleotide sequences across species or CpG hot spot. The L9X and R26X lead to the disruption of leader sequences, required for directing mitochondrial localization of the OTC precursor. Their phenotypes are severe, and neonatal onset. The G100R, R277W and V323M mutations were uniquely identified in patients with late onset OTC deficiency. The other genotypes are associated with neonatal onset. Out of 20 patients with OTC deficiency, only 6 patients are alive; two were liver transplanted, and normal in growth and development at 2, 4 years after transplantation respectively. Citrullinemia is an autosomal recessive disease, caused by the mutations in the argininosuccinate synthetase(ASS) gene. We identified in 3 major mutations in 11 unrelated Korean patients with citrullinemia; G324S, $IVS6^{-2}$ A to G, and 67 bp ins at nt 1125-1126. Among these, the 67 base pair insertion mutation is novel. The allele frequency of each mutation is; G324S(45%), IVS6-2 A to G(32%), and 67 base pair insertion(14%). All patients are diagnosed at neonatal or infantile age. Interestingly, two patients presented with stroke like episode. Out of 11 patients, 5 patients died. Among 6 patients alive, one patient was successfully liver transplanted.

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첨가제의 종류와 동해방지제의 농도에 따른 피조개, Scapharca broughtonii D형 유생의 냉동보존 효과 비교

  • Jo, Pil-Gue;Choi, Youn-Hee;Kang, Kyoung-Ho;Kho, Kang-Hee;Go, Chang-Soon;Kim, Byong-Hak;Lim, Han-Kyu;Choi, Cheol-Young;Chang, Young-Jin
    • The Korean Journal of Malacology
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    • v.18 no.2
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    • pp.77-82
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    • 2002
  • This study was performed to find out the optimal kind of additive and concentration of ethylene glycol (EG) as the cryoprotectant on the D-shaped larvae of arkshell, Scapharca broughtonii. The larvae in straws was carried in the programed freezer set uo at 0$^{\circ}C$, frozen to -12$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min, held for 10 minutes, seeded at -12$^{\circ}C$, finally refrozen to -35$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min and then directly plunged into liquid nitrogen. The survival of larvae frozen in 1.0 M and 2.0 M ethylene glycol added with 0.5 M sucrose were high (50.5 ${\pm}$ 1.3% and 51.9 ${\pm}$ 1.7%, respectively) in the early D-shaped larvae cryopreserved in 1 M and 2 M ethylene glycol diluted with 0.2 M and 0.5 M fructose, glucose and sucrose. In the late D-shaped larvae were cryopreserved according to five concentrations of ethylene glycol added with 0.5 M sucrose, the survival of larvae frozen in 2.0 M ethylene glycol was the highest as 51.9 ${\pm}$ 1.7%. The morphological differences in cells between unfrozen and frozen-thawed D-shaped larvae were not found by the scanning and transmission electron microscopy.

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Speciation of the Two Species of the Genus Hyla (Anura) in Korea (한국산 청개구리속 2조의 종분화에 관한 연구)

  • 양서영;박병상
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.11-20
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    • 1988
  • A comparison between two species of Hyla japonica and H. suweonesis in Korea was carried out to clarify their morphological, genetic and ecological differences, to estimate presumed divergent time between them, and to elucidate their probable process and mechanism of speciation. The results are as follows; 1.The breeding season of H. japonica was earlier than that of H. suweonensis about one and half months. 2.H.japonica was distributed all over the country, while H. suweonensis was restricted in Kyonggi plains. 3.On the extemal morphometric characters, differences were hardiy seen between two species. But on the discriminant function analysis, there was apparent differences between them. 4.Genic variation based on 24 loci showed that H. japonica populations were about two fold more variable than that of H. suweonensis populations. 5.Genetic relatedness between two species was rather remote (S =0.520, D=0.634) 6.Their presumed divergent time was estimated to be about three and a quarter mfllon years. It is presumed that H. suweonensis was originated from H. japonica during the period of late pleocene and early pleistocene. 7.Premating isolating mechanisrns (breeding season, mating call, and microhabitat segregation) may be operating to prevent gene ifow between them.

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Morphological and Genetic Characteristics of Colletotrichum gloeosporioides Isolated from Newly Emerging Static-Symptom Anthracnose in Apple

  • Jeon, Yongho;Cheon, Wonsu
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.34-34
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    • 2014
  • Filamentous fungi of the genus Colletotrichum (teleomorph, Glomerella) are considered major plant pathogens worldwide. Cereals, legumes, vegetables, and fruit trees may be seriously affected by this pathogen (1). Colletotrichum species cause typical disease symptoms known as anthracnoses, characterized by sunken necrotic tissue, where orange conidial masses are produced. Anthracnose appears in both developing and mature plant tissues (2). We investigated disease occurrence in apple orchards from 2013 to 2014 in northern Gyeongbuk province, Korea. Typical anthracnose with advanced symptoms was observed in all apple orchards studied. Of late, static fruit spot symptoms are being observed in apple orchards. A small lesion, which does not expand further and remains static until the harvesting season, is observed at the beginning of fruit growth period. In our study, static symptoms, together with the typical symptoms, were observed on apples. The isolated fungus was tested for pathogenicity on cv. 'Fuji apple' (fully ripe fruits, unripe fruits, and cross-section of fruits) by inoculating the fruits with a conidial suspension ($10^5$ conidia/ml). In apple inoculated with typical anthracnose fungus, the anthracnose symptoms progressed, and dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. However, in apple inoculated with fungi causing static symptoms, the size of the spots did not increase. Interestingly, the shape and size of the conidia and the shape of the appressoria of both types of fungi were found to be similar. The conidia of the two types of fungi were straight and cylindrical, with an obtuse apex. The culture and morphological characteristics of the conidia were similar to those of C. gloeosporioides (5). The conidia of C. gloeosporioides germinate and form appressoria in response to chemical signals such as host surface wax and the fruitripening hormone ethylene (3). In this study, the spores started to germinate 4 h after incubation with an ethephon suspension. Then, the germ tubes began to swell, and subsequently, differentiation into appressoria with dark thick walls was completed by 8 h. In advanced symptoms, fungal spores of virtually all the appressoria formed primary hyphae within 16 h. However, in the static-symptom fungus spores, no primary hyphae formed by 16 h. The two types of isolates exhibited different growth rates on medium containing apple pectin, Na polypectate, or glucose as the sole carbon. Static-symptom fungi had a >10% reduction in growth (apple pectin, 14.9%; Na polypectate, 27.7%; glucose, 10.4%). The fungal isolates were also genetically characterized by sequencing. ITS regions of rDNA, chitin synthase 1 (CHS1), actin (ACT), and ${\beta}$-tubulin (${\beta}t$) were amplified from isolates using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, ACT-512F and ACT-783R, and T1 and ${\beta}t2$ (5), respectively. The resulting sequences showed 100% identity with sequences of C. gloeosporioides at KC493156, and the sequence of the ${\beta}$t gene showed 100% identity with C. gloeosporioides at JX009557.1. Therefore, sequence data from the four loci studied proves that the isolated pathogen is C. gloeosporioides. We also performed random amplified polymorphic DNA-PCR, which showed clearly differentiated subgroups of C. gloeosporioides genotypes. The clustering of these groups was highly related to the symptom types of the individual strains.

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Changes in Oral Microbiota in Patients Receiving Radical Concurrent Chemoradiotherapy for The Head and Neck Squamous Cell Carcinoma

  • Kim, Jin Ho;Choi, Yoon Hee;An, Soo-Youn;Son, Hee Young;Choi, Chulwon;Kim, Seyeon;Chung, Jin;Na, Hee Sam
    • International Journal of Oral Biology
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    • v.43 no.1
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    • pp.13-21
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    • 2018
  • Radiotherapy (RT) is a mainstay in the treatment of head and neck squamous cell carcinoma (HNSCC). For locally advanced HCSCC, concurrent chemoradiotherapy (CCRT) benefits HCSCC patients in terms of better survival and loco-regional control. In this study, we evaluated changes in oral microbiota in patients, who received CCRT for head and neck cancer. Oral rinsed samples were weekly collected before and during CCRT and at 4 weeks following treatment from HNSCC patients, who had received 70 Gy of radiation delivered to the primary sites for over 7 weeks and concurrent chemotherapy. Oral microbiota changes in three patients were analyzed by next-generation sequencing using 16S rRNA 454 pyrosequencing. On an average, 15,000 partial 16S rRNA gene sequences were obtained from each sample. All sequences fell into 11 different bacterial phyla. During early CCRT, the microbial diversity gradually decreased. In a patient, who did not receive any antibiotics during the CCRT, Firmicutes and Proteobacteria were the most abundant phylum. During the early CCRT, proteobacteria gradually decreased while Firmicutes increased. During the late CCRT, firmicutes gradually decreased while Bacteroides and Fusobacteria increased. In all the patients, yellow complex showed a gradual decrease, while orange and red complex showed a gradual increase during the CCRT. At 4 weeks after CCRT, the recovery of oral microbiota diversity was limited. During CCRT, there was a gradual increase in major periodontopathogens in association with the deterioration of the oral hygiene. Henceforth, it is proposed that understanding oral microbiota shift should provide better information for the development of effective oral care programs for patients receiving CCRT for HNSCC.