• 제목/요약/키워드: protein tissues

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Sequence and Expression Analysis of c-fos Proto-oncogene in Korean Cattle (HANWOO) (한우 c-fos 유전자의 염기서열 및 발현분석)

  • Yu, S.L.;Chung, H.J.;Jung, K.C.;Lee, J.H.;Cho, K.W.;Choi, J.G.;Na, K.J.;Sang, B.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.891-900
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    • 2003
  • Cellular FOS(c-fos) protein is a transcription factor that forms heterodimers mostly with c-jun family and stimulates the transcription of genes containing AP-1 regulatory elements. This c-fos expression can control growth and differentiation of various precursor cells including myoblasts. The controls by c-fos gene have been identified for affecting skeletal muscle fiber traits which are the key determinants of meat quality in pigs. As a first step for identifying the relationship between c-fos gene and meat quality traits in cattle, we fully sequenced 1,443 bp of Hanwoo c-fos mRNA and analyzed expression patterns from various organs and muscle tissues. The sequence identities of Hanwoo c-fos with that of human, pig and mouse showed 89.8%, 93.3% and 87%, respectively. Analyses of the northern blot showed high c-fos expressions were obtained in spleen and rib muscle from 7 organs and 9 different parts of muscles investigated. These results presented here can be used as a valuable marker for meat quality related traits in cattle with further verification.

Optogenetics: a New Frontier for Cell Physiology Study (광유전학: 세포 생리 연구를 위한 새로운 frontier)

  • Byun, Jonghoe
    • Journal of Life Science
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    • v.25 no.8
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    • pp.953-959
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    • 2015
  • Optogenetics is the combination of optical and molecular strategies to control designated molecular and cellular activities in living tissues and cells using genetically encoded light-sensitive proteins. It involves the use of light to rapidly gate the membrane channels that allows for ion movement. Optogenetics began with the placing of light-sensitive proteins from green algae inside specific types of brain cells. The cells can then be turned on or off with pulses of blue and yellow light. Using the naturally occurring algal protein Channelrhodopsin-2 (ChR2), a rapidly gated light-sensitive cation channel, the number and frequency of action potentials can be controlled. The ChR2 provides a way to manipulate a single type of neuron while affecting no others, an unprecedented specificity. This technology allows the use of light to alter neural processing at the level of single spikes and synaptic events, yielding a widely applicable tool for neuroscientists and biomedical engineers. An improbable combination of green algae, lasers, gene therapy and fiber optics made it possible to map neural circuits deep inside the brain with a precision that has never been possible before. This will help identify the causes of disorders like depression, anxiety, schizophrenia, addiction, sleep disorder, and autism. Optogenetics could improve upon existing implanted devices that are used to treat Parkinson’s disease, obsessive-compulsive disorder and other ailments with pulses of electricity. An optogenetics device could hit more specific subsets of brain cells than those devices can. Applications of optogenetic tools in nonneuronal cells are on the rise.

Anti-Proliferative Effect of Ethanol on Normal and Cancer Cells (정상세포와 암세포의 증식 억제에 대한 에탄올의 영향)

  • Oh, Myung-Ju;Kim, Ji-Hyun;Park, Su-Hyun;Jeong, Young-Hwa;Wang, Kun;Cho, Byung-Wook;Jhun, Byung-H.
    • Journal of Life Science
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    • v.22 no.4
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    • pp.538-544
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    • 2012
  • Ethanol is known as being carcinogenic to humans. In addition, the anti-proliferative effects of ethanol have been described for a variety of tissues and cells. In this study, we investigated the anti-proliferative effects of ethanol on various cancer cells, particularly on oncogenic $ras$-transformed or-injected cells. Ethanol treatment inhibited the cell proliferation of normal control cells, but did not suppress the proliferation of various cancer cells and oncogenic $ras$-transformed cells. Furthermore, ethanol treatment did not interfere with DNA synthesis, which was induced by microinjecting the oncogenic $H-Ras^{V12}$ protein. The anti-proliferative effect of ethanol was rescued by antioxidants, such as $N$-acetylcysteine and 4-methlpyrazole. These results suggest that ethanol cytotoxicity is exerted through free radical formation, and that the anti-proliferative action site of ethanol cytotoxicity either lies upstream, or is independent of Ras.

Substitution of soybean meal with detoxified Jatropha curcas kernel meal: Effects on performance, nutrient utilization, and meat edibility of growing pigs

  • Li, Yang;Chen, Ling;Zhang, Yuhui;Wu, Jianmei;Lin, Yan;Fang, Zhengfeng;Che, Lianqiang;Xu, Shengyu;Wu, De
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.888-898
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    • 2018
  • Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

Effects of dietary supplementation of lipid-coated zinc oxide on intestinal mucosal morphology and expression of the genes associated with growth and immune function in weanling pigs

  • Song, Young Min;Kim, Myeong Hyeon;Kim, Ha Na;Jang, Insurk;Han, Jeong Hee;Fontamillas, Giselle Ann;Lee, Chul Young;Park, Byung-Chul
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.403-409
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    • 2018
  • Objective: The present study was conducted to investigate the effects of a lipid-coated zinc oxide (ZnO) supplement Shield Zn (SZ) at the sub-pharmacological concentration on intestinal morphology and gene expression in weanling pigs, with an aim to gain insights into the mechanism of actions for SZ. Methods: Forty 22-day-old weanling pigs were fed a nursery diet supplemented with 100 or 2,500 mg Zn/kg with uncoated ZnO (negative control [NC] or positive control [PC], respectively), 100, 200, or 400 mg Zn/kg with SZ for 14 days and their intestinal tissues were taken for histological and molecular biological examinations. The villus height (VH) and crypt depth (CD) of the intestinal mucosa were measured microscopically following preparation of the tissue specimen; expression of the genes associated with growth and immune function was determined using the real-time quantitative polymerase chain reaction. Results: There was no difference in daily gain, gain:feed, and diarrhea score between the SZ group and either of NC and PC. The VH and VH:CD ratio were less for the SZ group vs NC in the jejunum and duodenum, respectively (p<0.05). The jejunal mucosal mRNA levels of insulin-like growth factor (IGF-I) and interleukin (IL)-10 regressed and tended to regress (p = 0.053) on the SZ concentration with a positive coefficient, respectively, whereas the IL-6 mRNA level regressed on the SZ concentration with a negative coefficient. The mRNA levels of IGF-I, zonula occludens protein-1, tumor necrosis $factor-{\alpha}$, IL-6, and IL-10 did not differ between the SZ group and either of NC and PC; the occludin and transforming growth $factor-{\beta}1$ mRNA levels were lower for the SZ group than for PC. Conclusion: The present results are interpreted to suggest that dietary ZnO provided by SZ may play a role in intestinal mucosal growth and immune function by modulating the expression of IGF-I, IL-6, and IL-10 genes.

Overlapping Region of p53/Wrap53 Transcripts: Mutational Analysis and Sequence Similarity with microRNA-4732-5p

  • Pouladi, Nasser;Kouhsari, Shideh Montasser;Feizi, Mohammadali Hosseinpour;Gavgani, Reyhaneh Ravanbakhsh;Azarfam, Parvin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3503-3507
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    • 2013
  • Background: Although the majority of investigations concerned with TP53 and its protein have focused on coding regions, recently a set of studies highlighted significant roles of regulatory elements located in p53 mRNA, especially 5'UTR. The wrap53${\alpha}$ transcript is one of those that acts as a natural antisense agent, forming RNA-RNA hybrids with p53 mRNA and protecting it from degradation. Materials and Methods: In this study, we focused on the mutation status of exon $1{\alpha}$ of the WRAP53 gene (according to exon 1 of p53) in 160 breast tumor tissue samples and conducted a bioinformatics search for probable miRNA binding site in the p53/wrap53 overlapping region. Mutations were detected, using single stranded conformation polymorphism (SSCP) and sequencing. We applied the miRBase database for prediction of miRNAs which target overlapping region of p53/wrap53 transcripts. Results: Our results showed all samples to have wild type alleles in exon 1 of TP53 gene. We could detect a novel and unreported intronic mutation (IVS1+56, G>C) outside overlapping regions of p53/wrap53 genes in breast cancer tissues and also predict the presence of a binding site for miR-4732-5p in the 5'UTR of Wrap53 mRNA. Conclusions: From our findings we propose designing further studies focused on overexpression of miRNA-4732-5p and introducing different mutations in the overlapping region of wrap53 and p53 genes in order to study their effects on p53 and its ${\Delta}N$ isoform (${\Delta}$40p53) expression. The results may provide new pieces in the p53 targeting puzzle for cancer therapy.

Molecular Characterization and Expression Analysis of a Glutathione S-Transferase cDNA from Abalone (Haliotis discus hannai) (북방전복 (Haliotis discus hannai)에서 분리한 Glutathione S-transferase 유전자의 분자생물학적 고찰 및 발현분석)

  • Moon, Ji Young;Park, Eun Hee;Kong, Hee Jeong;Kim, Dong-Gyun;Kim, Young-Ok;Kim, Woo-Jin;An, Cheul Min;Nam, Bo-Hye
    • The Korean Journal of Malacology
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    • v.30 no.4
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    • pp.399-408
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    • 2014
  • Glutathione S-transferases (GSTs) are a superfamily of detoxification enzymes that primarily catalyze the nucleophilic addition of reduced glutathione to both endogenous and exogenous electrophiles. In this study, we isolated and characterized a full-length of alpha class GST cDNA from the abalone (Haliotis discus hannai). The abalone GST cDNA encodes a 223-amino acid polypeptide with a calculated molecular mass of 25.8 kDa and isoelectric point of 5.69. Multiple alignments and phylogenetic analysis with the deduced abalone GST protein revealed that it belongs to the alpha class GSTs and showed strong homology with disk abalone (Haliotis discus discus) putative alpha class GST. Abalone GST mRNA was ubiquitously detected in all tested tissues. GST mRNA expression was comparatively high in the mantle, gill, liver, and digestive duct, however, lowest in the hemocytes. Expression level of abalone GST mRNA in the mantle, gill, liver, and digestive duct was 182.7-fold, 114.8-fold, 4675.8-fold, 406.1-fold higher than in the hemocytes, respectively. Expression level of abalone GST mRNA in the liver was peaked at 6 h post-infection with Vibrio parahemolyticus and decreased at 12 h post-infection. While the expression level of abalone GST mRNA in the hemocytes was drastically increased at 3 h post-infection with Vibrio parahemolyticus. These results suggest that abalone GST is conserved through evolution and may play roles similar to its mammalian counterparts.

H9 Inhibits Tumor Growth and Induces Apoptosis via Intrinsic and Extrinsic Signaling Pathway in Human Non-Small Cell Lung Cancer Xenografts

  • Kim, Min-Je;Kwon, Sae-Bom;Ham, Seung Hoon;Jeong, Eui-Suk;Choi, Yang-Kyu;Choi, Kang Duk;Hong, Jin Tae;Jung, Seung Hyun;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.648-657
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    • 2015
  • H9, a novel herbal extract, demonstrated cytotoxicity in A549 non-small cell lung cancer (NSCLC) cell lines. In this study, we investigated whether H9, and/or co-treatment with an anticancer drug, pemetrexed (PEM), inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. The mice were separated into groups and administered H9 and PEM for 2 weeks. Protein and mRNA levels were detected using western blotting and reverse transcription polymerase chain reaction, respectively; immunohistochemistry (IHC) was also performed on the tumor tissues. H9 and co-treatment with PEM induced the cleavage of proapoptotic factors, such as caspase-3, caspase-8, caspase-9, and poly(ADP)-ribose polymerase (PARP). Expression levels of cell-death receptors involving Fas/FasL, TNF-related apoptosisinducing ligands (TRAIL), and TRAIL receptors were increased by H9 and co-treatment with PEM. Furthermore, analysis of levels of cell-cycle modulating proteins indicated that tumor cells were arrested in the G1/S phase. In addition, the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt survival signaling pathways were inhibited by H9 and co-treatment with PEM. In conclusion, H9 and co-treatment with PEM inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. These results indicate that H9 and co-treatment with PEM can be used as an anticancer therapy in NSCLC.

Identification and characterization of the phytocystatin family from Brassica rapa

  • Hong, Joon-Ki;Hwang, Jung-Eun;Park, Tae-Ho;Zang, Yun-Xiang;Lee, Sang-Choon;Kwon, Soo-Jin;Mun, Jeong-Hwan;Kim, Hyun-Uk;Kim, Jin-A;Jin, Mi-Na;Kim, Jung-Sun;Lee, Soo-In;Lim, Myung-Ho
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.317-327
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    • 2008
  • Phytocystatins, which are inhibitors of plant cysteine peptidases, are involved in the regulation of protein turnover and in the defense against insect pests and pathogens. Extensive searches in the Brassica rapa genome allowed the prediction of at least eight different phytocystatin genes on seven chromosomes in the B. rapa genome. Structure comparisons based on alignments of the all BrCYS ($\underline{B}$. $\underline{r}apa$ $phyto{\underline{cys}}tatin$) proteins using the CLUSTALW program revealed conservation of the three consensus motifs known to interact with the active site of cysteine peptidases. According to the phylogenetic analysis based on the deduced amino acid sequences, the eight BrCYS proteins were divided into several clusters related to the orthologous phytocystatin. The predicted three-dimensional structure models of the eight BrCYS proteins demonstrate that all of these proteins are similar to the reported crystal structure of oryzacystatin-I (OC-I). Digital northern and RT-PCR analyses indicated that the eight BrCYS genes exhibit different expression patterns in B. rapa tissues and respond differently to abiotic stimuli. The differences in gene structure and expression between the eight BrCYS genes suggest that these proteins may play diverse physiological roles in B. rapa and may interact with cysteine peptidases through different mechanisms.

The Effect of PDGF-Loaded Biodegradable Membrane on Early Healing Stage in Guided Tissue Regeneration (흡수성 차폐막의 치주조직 재생에 혈소판유래 성장인자가 미치는 영향)

  • Rhyu, In-Chul;Bae, Kyoo-Hyun;Seol, Yang-Jo;Ku, Young;Lee, Seung-Jin;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.507-519
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    • 1999
  • The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

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