• Title/Summary/Keyword: PCR condition

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Expression Patterns of Growth Related Genes in Juvenile Red Spotted Grouper (Epinephelus akaara) with Different Growth Performance after Size Grading

  • Mun, Seong Hee;You, Jin Ho;Oh, Hyeon Ji;Lee, Chi Hoon;Baek, Hea Ja;Lee, Young-Don;Kwon, Joon Yeong
    • Development and Reproduction
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    • v.23 no.1
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    • pp.35-42
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    • 2019
  • Fish shows great difference in growth rate between individuals during larval development and early growth. This difference seriously reduces the production efficiency in fish culture. Growth hormone (GH)/Insulin-like growth factor 1 (IGF1) system is said to play some pivotal roles in fish growth. In this study, we investigated differences of GH, IGF1 and GHR gene expressions in juvenile red spotted grouper (Epinephelus akaara) with different growth performance. Red spotted groupers were reared under the same environmental condition (water temperature $24{\pm}1^{\circ}C$, natural light) for 96 days after hatching. They were divided into 3 groups by size (fast growing, middle growing and slow growing groups: FGG, MGG, and SGG, respectively). RNA was extracted from the brain, liver and muscle tissues from each group, and target gene expression was examined by real-time PCR. In the brain with pituitary gland, expression of GH gene in FGG was significantly higher than the expression in SGG, but the expression of IGF1 and GHR genes in the muscle was highest in SGG. Difference of GHR and IGF1 mRNA in the liver between groups with different growth performance was less clear than that in other tissues, although level of IGF1 mRNA was higher in SGG than in MGG. These results suggest that hormonal governing of growth is not the same in fast growing and slow growing fish, and size grading could cause a shift of hormonal state and growth pattern in this species.

Effect of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia

  • Zhao, Chuan;Guo, Yushu;Wang, Ruoxi;Cheng, Cheng;Chen, Xiangmei
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.6
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    • pp.517-523
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    • 2021
  • The present study aims to investigate the impact of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia. Thirty Sprague-Dawley rats were randomly divided into five groups, including normal diet group, and diabetes model (DM) group, DM + metformin treatment (DMM) group, DMM + hypoxia treatment (DMMH) group and DMMH + hydrogen-rich water (DMMHR) group. We found that the levels of lactic acid, pyruvate and lactate dehydrogenase were significantly lower in the blood of DMMHR group than DMMH group. Superoxide dismutase and glutathione levels in liver and heart were significantly higher in DMMH group after hydrogen-rich water treatment, while malondialdehyde and oxidized glutathione levels were decreased in DMMHR group when compared with DMMH group, which indicates that hydrogen-rich water could reduce oxidative stress. qPCR analysis demonstrated that that pro-apoptotic genes Bax/Caspase-3 were upregulated in DM group and metformin treatment suppressed their upregulation (DMM group). However, hypoxic condition reversed the effect of metformin on apoptotic gene expression, and hydrogen-rich water showed little effect on these genes under hypoxia. HE staining showed that hydrogen-rich water prevented myocardial fiber damages under hypoxia. In summary, we conclude that hydrogen-rich water could prevent lactate accumulation and reduce oxidant stress in diabetic rat model to prevent hypoxia-induced damages. It could be served as a potential agent for diabetes patients with metformin treatment to prevent lactic acidosis and reduce myocardial damages under hypoxic conditions.

Hyperglycemia aggravates decrease in alpha-synuclein expression in a middle cerebral artery occlusion model

  • Kang, Ju-Bin;Kim, Dong-Kyun;Park, Dong-Ju;Shah, Murad-Ali;Kim, Myeong-Ok;Jung, Eun-Jung;Lee, Han-Shin;Koh, Phil-Ok
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.195-202
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    • 2018
  • Hyperglycemia is one of the major risk factors for stroke. Hyperglycemia can lead to a more extensive infarct volume, aggravate neuronal damage after cerebral ischemia. ${\alpha}$-Synuclein is especially abundant in neuronal tissue, where it underlies the etiopathology of several neurodegenerative diseases. This study investigated whether hyperglycemic conditions regulate the expression of ${\alpha}$-synuclein in middle cerebral artery occlusion (MCAO)-induced cerebral ischemic injury. Male Sprague-Dawley rats were treated with streptozotocin (40 mg/kg) via intraperitoneal injection to induce hyperglycemic conditions. MCAO were performed four weeks after streptozotocin injection to induce focal cerebral ischemia, and cerebral cortex tissues were obtained 24 hours after MCAO. We confirmed that MCAO induced neurological functional deficits and cerebral infarction, and these changes were more extensive in diabetic animals compared to non-diabetic animals. Moreover, we identified a decrease in ${\alpha}$-synuclein after MCAO injury. Diabetic animals showed a more serious decrease in ${\alpha}$-synuclein than non-diabetic animals. Western blot and reverse-transcription PCR analyses confirmed more extensive decreases in ${\alpha}$-synuclein expression in MCAO-injured animals with diabetic condition than these of non-diabetic animals. It is accepted that ${\alpha}$-synuclein modulates neuronal cell death and exerts a neuroprotective effect. Thus, the results of this study suggest that hyperglycemic conditions cause more serious brain damage in ischemic brain injuries by decreasing ${\alpha}$-synuclein expression.

Alteration of Apoptosis during Differentiation in Human Dental Pulp-Derived Mesenchymal Stem Cell

  • Lee, Hyeon-Jeong;Park, Byung-Joon;Jeon, Ryoung-Hoon;Jang, Si-Jung;Son, Young-Bum;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.2-9
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    • 2019
  • Because mesenchymal stem cells (MSCs) maintain distinct capacities with respect to self-renewal, differentiation ability and immunomodulatory function, they have been highly considered as the therapeutic agents for cell-based clinical application. Of particular, differentiation condition alters characteristics of MSCs, including cellular morphology, expression of gene/protein and cell surface molecule, immunological property and apoptosis. However, the previous results for differentiation-related apoptosis in MSCs have still remained controversial due to varied outcomes. Therefore, the present study aimed to disclose periodical alterations of pro- and anti-apoptosis in MSCs under differentiation inductions. The human dental pulp-derived MSCs (DP-MSCs) were differentiated into adipocytes and osteoblasts during early (1 week), middle (2 weeks) and late (3 weeks) stages, and were investigated on their apoptosis-related changes by Annexin V assay, qRT-PCR and western blotting. The ratio of apoptotic cell population was significantly (p < 0.05) elevated during the early to middle stages of differentiations but recovered up to the similar level of undifferentiated state at the late stage of differentiation. In the expression of mRNA and protein, whereas expressions of pro-apoptosis-related makers (BAX and BAK) were not altered in any kind and duration of differentiation inductions, anti-apoptosis marker (BCL2) was significantly (p < 0.05) elevated even at the early stage of differentiations. The recovery of apoptotic cell population at the late stage of differentiation is expected to be associated with the response by elevation of anti-apoptotic molecules. The present study may contribute on understanding for cellular mechanism in differentiation of MSCs and provide background data in clinical application of MSCs in the animal biotechnology to develop effective and safe therapeutic strategy.

Isolation and Identification of Alkali-tolerant Bacteria from Near-Shore Soils in Dokdo Island

  • Namirimu, Teddy;Kim, Jinnam;Zo, Young-Gun
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.105-115
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    • 2019
  • Saline or alkaline condition in soil inhibits growth of most crop plants and limits crop yields in many parts of the world. Augmenting an alkaline soil with alkali-tolerant bacteria capable of promoting plant growth can be a promising approach in expanding fertile agricultural land. Near-shore environments of Dokdo Island, a remote island located in the middle of the East Sea, appear to have patches of seawater-influenced haloalkaline soil that is unsupportive for growth of conventional plants. To exploit metabolic capacities of alkali-tolerant bacteria for promoting plant growth in saline or alkaline soils, we isolated of alkali-tolerant bacteria from near-shore soil samples in Dokdo and investigated properties of the isolates. Alkali-tolerant bacteria were selectively cultivated by inoculating suspended and diluted soil samples on a plate medium adjusted to pH 10. Fifty colonies were identified based on their $GTG_5$-PCR genomic fingerprints and 16S rRNA gene sequences. Most isolates were affiliated to alkali-tolerant and/or halotolerant genera or species of the phyla Firmicutes (68%), Proteobacteria (30%) and Actinobacteria (2%). Unlike the typical soil bacterial flora in the island, alkali-tolerant isolates belonged to only certain taxa of terrestrial origin under the three phyla, which have traits of plant growth promoting activities including detoxification, phytohormone production, disease/pest control, nitrogen-fixation, phosphate solubilization or siderophore production. However, Firmicutes of marine origin generally dominated the alkali-tolerant community. Results of this study suggest that haloalkaline environments like Dokdo shore soils are important sources for plant growth promoting bacteria that can be employed in bio-augmentation of vegetation-poor alkaline soils.

Investigation of effect of zirconia on osseointegration by surface treatments (지르코니아 표면처리가 골유착에 미치는 영향)

  • Jeong, Jin-Woo;Song, Young-Gyun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.37 no.1
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    • pp.23-30
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    • 2021
  • Purpose: The aim of this study was to investigate effect of zirconia on osseointegration and Surface appearance by surface treatments using various acid solution. Materials and Methods: The prepared zirconia disks were treated with hydrofluoric acid solution and photo-assisted etching under various condition. The surface was analyzed by SEM and the surface roughness was analyzed by using surface profiler. The osteogenic effect of MC3T3-E1 cells was assessed via fluorescent staining observation and reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Various roughness were obtained according to the surface treatment method. The surface roughness increased in the group treated with hydrofluoric acid solution, but that had week network structure. In the method using photo-assisted etching, the surface roughness increased in micro units. Cell reaction showed better results in the photo-assisted etching group than in the hydrofluoric acid-treated group (P < 0.05). And it showed even osteoblastic cell distribution in photo-assisted etching group. Conclusion: As a result, the photo-assisted etching method is more effective than the simple acid solution treatment for zirconia treatment for osseointegration.

The Effects of steam heat processing of Helianthus tuberosi Rhizoma on Blood glucose lowering (국우(菊芋) 증자가 혈당강하작용에 미치는 영향)

  • Kim, Jin-Woo;Ha, Mi-Ae;Shin, Yong-Wook
    • The Korea Journal of Herbology
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    • v.32 no.5
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    • pp.39-46
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    • 2017
  • Objective : This study was designed to evaluate the hypoglycemic effects of Helianthus tuberosi Rhizoma extracts and its optimum Heat processing conditions Methods : We investigated the Salivary ${\alpha}$-amylase, pancreas ${\alpha}$-amylase and ${\alpha}$-glucosidase inhibitory activities of extracts from Steam Heated Helianthus tuberosi Rhizoma Ext. The inhibitory activities of a 50% EtOH extract of Steam Heated Helianthus tuberosi Rhizoma Ext against ${\alpha}$-glucosidases were evaluated in this study. Inhibiting these enzymes involved in the absorption of disaccharides significantly decreases the postprandial increase in blood glucose level after a mixed carbohydrate diet. Furthermore, the postprandial blood glucose lowering effect of Steam Heated Helianthus tuberosi Rhizoma Ext. was compared to a known type 2 diabetes drug(Acarbose(R)) in a mice model. Steam Heated Helianthus tuberosus L. Ext significantly reduced the blood glucose increase after glucose loading. Results : The results were confirmed by real-time PCR that after treated with Streptozotocin in L6 cells, induced expression of GLUT4, after the steamed Helianthus tuberosus L. Ext. treated, observed its expression was increased. Steam Heated Helianthus tuberosus L Ext treated 4 hours in L6 cells, cytotoxicity was measured in MTT assay. Its toxicity were 5.7%, 9% and 11.3% at the treatment concentration $12.5{\mu}g/m{\ell}$, $25{\mu}g/m{\ell}$, the $50{\mu}g/m{\ell}$ respectively. Conclusions : Overall, the results of this study indicate that Hypoglycemic effect of Helianthus tuberosi Rhizoma caused by the Steam heat treatment, the optimum Heat processing condition is steamming at $121^{\circ}C$ for 30 min, and it will provide the basis for developing a useful dietary supplement for controlling postprandial hyperglycemia.

Development of transgenic disease-resistance root stock for growth of watermelon.(oral)

  • S.M. Cho;Kim, J.Y.;J.E. Jung;S.J. Mun;S.J. Jung;Kim, K.S.;Kim, Y.C.;B.H. Cho
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.65.2-65
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    • 2003
  • To protect the plant against several soil-borne pathogens, we are currently constructing disease-resistant transgenic root stock for the growth of cucurbitaceae vegetable plants, watermelon and gourd. We made a watermelon cDNA library from Cladosporium cucumerinum-Infected leaves for substractive hybriazation and differential screening. We isolated the several pathogen inducible cDNA clones, such as caffeoyl-CoA-methyltransferase, LAA induced protein, receptor-like kinase homolog, hydroxyproline-rich glycoprotein, catalase, calmodulin binding protein, mitochondrial ATPase beta subunit, methyl tRNA synthetase and WRKY transcription factors. We previously obtained CaMADS in pepper and galactinol synthase ( CsGolS) in cucumber that were confirmed to be related with disease-resistance. CaMADS and CsGolS2 were transformed into the inbred line 'GO701-2' gourd, the inbred line '6-2-2' watermelon and the Kong-dye watermelon by Agrobacterium tumerfaciens LBA4404. Plant growth regulators (zeatin, BAP and IAA) were used for shoot regeneration and root induction for optimal condition. Putative transgenic plants were selected in medium containing 100mg/L kanamycin and integration of the CaMADS and CsGO/S2 into the genomic DNA were demonstrated by the PCR analysis. We isolated major soil-borne pathogens, such as Monosporascus cannonballus, Didymella bryoniae, Cladosporium cuvumerinum from the cultivation area of watermelon or root stock, and successfully established artificial inoculation method for each pathogen. This work was supported by a grant from BioGreen 21 program, Rural Development Administration, Republic of Korea.

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DNA Array Analysis of Changes in Gene Expression Profile in DHEA-induced PCO

  • Yu, Jeong-Min;Yoo, Seong-Jin;Kim, Do-Rim;Youm, Mi-Young, Kim, Jee-Yun;Kang, Sung-Goo
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.112-112
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    • 2002
  • Under normal conditions, women produce a single dominant follicle that participates in a single ovuation each menstrual cycle. But Polycystic ovary syndrome(PCOS) conditions, folliculogenesis does not proceed normally. This condition leads to the accumlation of large numbers of small graffian follicles in which the theca interstitial cells (TIC) produce abnormally large amounts of androgen. PCOS is probably the most common endocrine disorder, affecting women of reprodutive age with 5-10% prevalence estimate. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries are clinical hallmarks of women with PCOS. Its etiology remains unknown. To investigate the gene expression pattern of ovary in PCO-induced rat, we used cDNA expression analysis. Total RNA was extracted from the ovary of PCO-induced rat and reverse-transcribed in the presence of[$\alpha$$^{32}$P]-dATP Which were hybridized to Atlas$^{TM}$ Rat Toxicology 1.2 array (Clontech) representing approximately 1176 rat genes. We compared gene expression between ovary of pco-induced immature female rats and control. Differential gene expression profiles were revealed (LIFR-alpha, ADRA1A, Heat shock 90-kDa protein A, PDGFRA). Reverse transcription-polymerase chain reaction(RT-PCR) was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the altered expression of genes and PCO is a matter of further investigation. This study was supported by Korea Science and Engineering Foundation(KOSEF)

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Ginsenoside Re prevents 3-methyladenine-induced catagen phase acceleration by regulating Wnt/β-catenin signaling in human dermal papilla cells

  • Gyusang Jeong;Seung Hyun Shin;Su Na Kim;Yongjoo Na;Byung Cheol Park;Jeong Hun Cho;Won-Seok Park;Hyoung-June Kim
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.440-447
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    • 2023
  • Background: The human hair follicle undergoes cyclic phases-anagen, catagen, and telogen-throughout its lifetime. This cyclic transition has been studied as a target for treating hair loss. Recently, correlation between the inhibition of autophagy and acceleration of the catagen phase in human hair follicles was investigated. However, the role of autophagy in human dermal papilla cells (hDPCs), which is involved in the development and growth of hair follicles, is not known. We hypothesized that acceleration of hair catagen phase upon inhibition of autophagy is due to the downregulation of Wnt/β-catenin signaling in hDPCs, and that components of Panax ginseng extract can increase the autophagic flux in hDPCs. Methods: We generated an autophagy-inhibited condition using 3-methyladenine (3-MA), a specific autophagy inhibitor, and investigated the regulation of Wnt/β-catenin signaling using the luciferase reporter assay, qRT-PCR, and western blot analysis. In addition, cells were cotreated with ginsenoside Re and 3-MA and their roles in inhibiting autophagosome formation were investigated. Results: We found that the unstimulated anagen phase dermal papilla region expressed the autophagy marker, LC3. Transcription of Wnt-related genes and nuclear translocation of β-catenin were reduced after treatment of hDPCs with 3-MA. In addition, treatment with the combination of ginsenoside Re and 3-MA changed the Wnt activity and hair cycle by restoring autophagy. Conclusions: Our results suggest that autophagy inhibition in hDPCs accelerates the catagen phase by downregulating Wnt/β-catenin signaling. Furthermore, ginsenoside Re, which increased autophagy in hDPCs, could be useful for reducing hair loss caused by abnormal inhibition of autophagy.