• Title/Summary/Keyword: Essential Amino Acid

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Induction of Midbrain Dopaminergic Phenotype in Nurr 1-Over expressing Human Neural Stem Cells (사람 신경 간세포에서 도파민 신경세포 분화유도에 대한 Nurr 1 유전자의 역할 규명)

  • Kim, Han-Jip;Lee, Haksup;Kim, Hyon-Chang;Min, Churl-Ki;Lee, Myung-Ae;Kim, Seung-Up;Han, Jin;Youm, Jae-Boum;Kim, Nari;Park, Won, Sun;Kim, Taeho;Kim, Euiyong;Han, Il-Yong
    • KSBB Journal
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    • v.20 no.5 s.94
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    • pp.363-370
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    • 2005
  • Neural stem cells (NSCs) of the central nervous system (CNS) have raised a great interest not only for their importance in basic neural development but also for their therapeutic potentials in neurologically degenerative diseases such as Parkinson's, Alzheimer and stroke. During the CNS development, two molecular cascades determine specification of midbrain dopamine system. In one pathway, FGF-8, sonic hedgehog and transcription factor Nurr1 specify dopamine neurotransmitter phenotype. In the other, transcription factors $Lm{\times}lb\;and\;Pt{\times}3$ are required for induction of dopaminergic neurons. In Nurr1 knockout mouse, tyrosine hydroxylase (TH) positive cells fail to appear in substantia nigra, indicating that Nurr1 is essential in specification of dopaminergic cell phenotype. In this study, we used the immortalized human NSCs retrovirally transduced with Nurr1 gene to probe the Nurr1 mediated mechanism to induce dopamine phenotype. While Nurr1 over-expression alone did not generate dopamine phenotype in NSCs, applications of retinoid and forskolin induced expression of TH and AADC mRNAs. In addition, co-cultures of Nurr1 expressing NSCs with human astrocytes induced a marked increase of TH expression. In this co-culture system, the addition of retinoid and forskolin dramatically increased expression of TH. These results indicate that the immortalized human NSCs with Nurr1 gene could have a clinical utility for cell replacement for the Parkinson patients.

Partial Interferon-γ Receptor Deficiency in Patients with Disseminated Tuberculosis (파종성 결핵 환자에서 interferon-γ 수용체의 부분결핍에 관한 연구)

  • Hwang, Jung Hye;Koh, Won-Jung;Lee, Shin Hye;Kim, Eun Joo;Kang, Eun Hae;Suh, Gee Young;Chung, Man Pyo;Kim, Hojoong;Kwon, O Jung
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.11-17
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    • 2005
  • Background : Interferon-gamma ($IFN-{\gamma}$) is essential in the immune response to mycobacterial infections, and a complete or partial deficiency in the $IFN-{\gamma}$ receptor 1 ($IFN{\gamma}R1$) or the $IFN-{\gamma}$ receptor 2 ($IFN{\gamma}R2$) have been reported to confer susceptibility to a disseminated infection with nontuberculous mycobacteria. However, similar mutations in the $IFN-{\gamma}$ receptor have not been specifically examined in the patients with clinical tuberculosis. Methods : This study searched for mutations in the $IFN-{\gamma}$ receptor gene that resulted in a partial $IFN-{\gamma}$ receptor deficiency in six patients with disseminated tuberculosis. The previously identified $IFN{\gamma}R1$ and $IFN{\gamma}R2$ coding regions were sequenced after amplification. Results : There was no partial $IFN{\gamma}R1$ deficiency including a homozygous recessive missense mutation causing an amino-acid substitution in the extracellular domain of the receptor (I87T) and a hotspot for small deletions (818delT, 818del4, 818insA) found in any of the patients. In addition, a partial $IFN{\gamma}R2$ deficiency of the homozygous missense mutation (R114C) was not found in any of the patients. Conclusion : Genetic defects causing a partial $IFN-{\gamma}$ receptor deficiency were not identified in our patients with disseminated tuberculosis.

Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs) (철 저장 단백질 ferritin과 kinesin 1 결합 규명)

  • Jang, Won Hee;Jeong, Young Joo;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.6
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    • pp.698-704
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    • 2016
  • The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

Characterization of Physiological Properties in Vibrio fluvialis by the Deletion of Oligopeptide Permease (oppA) Gene (Vibrio fluvialis oligopeptide permease (oppA) 유전자 deletion에 의한 생리적 특성)

  • Ahn Sun Hee;Lee Eun Mi;Kim Dong Gyun;Hong Gyoung Eun;Park Eun Mi;Kong In Soo
    • Journal of Life Science
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    • v.16 no.1
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    • pp.131-135
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    • 2006
  • Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.

Roles of the Insulin-like Growth Factor System in the Reproductive Function;Uterine Connection (Insulin-like Growth Factor Systems의 생식기능에서의 역할;자궁편)

  • Lee, Chul-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.247-268
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    • 1996
  • It has been known for a long time that gonadotropins and steroid hormones play a pivotal role in a series of reproductive biological phenomena including the maturation of ovarian follicles and oocytes, ovulation and implantation, maintenance of pregnancy and fetal growth & development, parturition and mammary development and lactation. Recent investigations, however, have elucidated that in addition to these classic hormones, multiple growth factors also are involved in these phenomena. Most growth factors in reproductive organs mediate the actions of gonadotropins and steroid hormones or synergize with them in an autocrine/paracrine manner. The insulin-like growth factor(IGF) system, which is one of the most actively investigated areas lately in the reproductive organs, has been found to have important roles in a wide gamut of reproductive phenomena. In the present communication, published literature pertaining to the intrauterine IGF system will be reviewed preceded by general information of the IGF system. The IGF family comprises of IGF-I & IGF-II ligands, two types of IGF receptors and six classes of IGF-binding proteins(IGFBPs) that are known to date. IGF-I and IGF-II peptides, which are structurally homologous to proinsulin, possess the insulin-like activity including the stimulatory effect of glucose and amino acid transport. Besides, IGFs as mitogens stimulate cell division, and also play a role in cellular differentiation and functions in a variety of cell lines. IGFs are expressed mainly in the liver and messenchymal cells, and act on almost all types of tissues in an autocrine/paracrine as well as endocrine mode. There are two types of IGF receptors. Type I IGF receptors, which are tyrosine kinase receptors having high-affinity for IGF-I and IGF-II, mediate almost all the IGF actions that are described above. Type II IGF receptors or IGF-II/mannose-6-phosphate receptors have two distinct binding sites; the IGF-II binding site exhibits a high affinity only for IGF-II. The principal role of the type II IGF receptor is to destroy IGF-II by targeting the ligand to the lysosome. IGFs in biological fluids are mostly bound to IGFBP. IGFBPs, in general, are IGF storage/carrier proteins or modulators of IGF actions; however, as for distinct roles for individual IGFBPs, only limited information is available. IGFBPs inhibit IGF actions under most in vitro situations, seemingly because affinities of IGFBPs for IGFs are greater than those of IGF receptors. How IGF is released from IGFBP to reach IGF receptors is not known; however, various IGFBP protease activities that are present in blood and interstitial fluids are believed to play an important role in the process of IGF release from the IGFBP. According to latest reports, there is evidence that under certain in vitro circumstances, IGFBP-1, -3, -5 have their own biological activities independent of the IGF. This may add another dimension of complexity of the already complicated IGF system. Messenger ribonucleic acids and proteins of the IGF family members are expressed in the uterine tissue and conceptus of the primates, rodents and farm animals to play important roles in growth and development of the uterus and fetus. Expression of the uterine IGF system is regulated by gonadal hormones and local regulatory substances with temporal and spatial specificities. Locally expressed IGFs and IGFBPs act on the uterine tissue in an autocrine/paracrine manner, or are secreted into the uterine lumen to participate in conceptus growth and development. Conceptus also expresses the IGF system beginning from the peri-implantation period. When an IGF family member is expressed in the conceptus, however, is determined by the presence or absence of maternally inherited mRNAs, genetic programming of the conceptus itself and an interaction with the maternal tissue. The site of IGF action also follows temporal (physiological status) and spatial specificities. These facts that expression of the IGF system is temporally and spatially regulated support indirectly a hypothesis that IGFs play a role in conceptus growth and development. Uterine and conceptus-derived IGFs stimulate cell division and differentiation, glucose and amino acid transport, general protein synthesis and the biosynthesis of mammotropic hormones including placental lactogen and prolactin, and also play a role in steroidogenesis. The suggested role for IGFs in conceptus growth and development has been proven by the result of IGF-I, IGF-II or IGF receptor gene disruption(targeting) of murine embryos by the homologous recombination technique. Mice carrying a null mutation for IGF-I and/or IGF-II or type I IGF receptor undergo delayed prenatal and postnatal growth and development with 30-60% normal weights at birth. Moreover, mice lacking the type I IGF receptor or IGF-I plus IGF-II die soon after birth. Intrauterine IGFBPs generally are believed to sequester IGF ligands within the uterus or to play a role of negative regulators of IGF actions by inhibiting IGF binding to cognate receptors. However, when it is taken into account that IGFBP-1 is expressed and secreted in primate uteri in amounts assessedly far exceeding those of local IGFs and that IGFBP-1 is one of the major secretory proteins of the primate decidua, the possibility that this IGFBP may have its own biological activity independent of IGF cannot be excluded. Evidently, elucidating the exact role of each IGFBP is an essential step into understanding the whole IGF system. As such, further research in this area is awaited with a lot of anticipation and attention.

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Methylenetetrahydrofolate Reductase C677T Polymorphism in Gastric Cancer (위암에서 Methylenetetrahydrofolate Reductase C677T의 유전자 다형성)

  • Seo Won;Park Won Cheol;Lee Jeong Kyun;Kim Jeong Jung
    • Journal of Gastric Cancer
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    • v.5 no.1
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    • pp.10-15
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    • 2005
  • Purpose: Recently the role of vitamins, folate in particular, has been emphasized in the maintenance of health. Folate deficiency is known to give rise to developmental delay, immature vascular disease, neural tube defect, acute leukemia, atherosclerotic vascular disease, delivery defects, breast cancer, and particularly gastrointestinal neoplasia. Methylenetetrahydrofolate reductase (MTHFR) is an essential enzyme in folate metaboism, and influences DNA synthesis and DNA methylation. Generally, folate deficiency is associated with gastrointestinal neoplasms. The amino-acid- changing and enzyme-activity-reducing nucleotide polymorphism (766C$\rightarrow$T/ Ala222Val) has been described in the MTHFR polymorphism and leads to low enzyme activity that may reduce the capacity of DNA methylation and possibly uracil mis-incorporation into DNA. These processes may be critical in the oncogenic transformation of human cells, especially in colorectal carcinomas. We investigated the relationship between the MTHFR polymorphism in gastric cancer and the tumor site, the smoking history, and the alcoholic history. Materials and Methods: Ninety-six (96) individuals with gastric cancer and 287 healthy persons were analyzed. Blood sampling was performed, PCR-RFLP was analyzed, and MTHFR polymorphism genotypes of C/C, C/T, and T/T were obtained and analyzed statistically for their correlation. Results: In the gastric cancer group there were 69 ($72\%$) males and 27 ($28\%$) females. There were also 58 cases ($60\%$) involving the gastric lower body, 20 cases ($21\%$) the gastric mid-body, and 18 cases ($19\%$) the gastric upper body. In the control group there were 169 ($59\%$) males and 118 ($41\%$) females. Among the gastric cancer, 56 ($61\%$) smoking patients, 40 ($39\%$) non-smoking patients, 45($47\%$) alcoholic patients, 51 ($53\%$) non-alcoholic patients. In the gastric cancer group, MTHER polymorphisms were C/C in 18 ($19\%$) cases, C/T in 59 ($61\%$) cases, T/T in 19 ($20\%$) cases. In the control group polymorphisms were C/C 116 ($40\%$) cases, C/T 103 ($36\%$) cases, and T/T 68 ($24\%$) cases (P=0.045). In cases of lower gastric body cancer, polymorphisms were C/C in 16 ($24\%$) C/C in 16 ($24\%$) cases and C/T or T/T in 42 ($72\%$) cases. In cases of upper and mid-body cancer, polymorphisms were C/C in 2 ($5\%$) cases and C/T or T/T 36 ($95\%$) cases (P=0.006). In the non-smoking patient group, polymorphisms were C/C in 5 (12%) cases and C/T or T/T in 35 ($88\%$) cases. In the smoking patient group, C/C in 13 ($23\%$) cases and C/T or T/T in 43 ($77\%$) cases (P=0.189). In the non-alcoholic patient group, polymorphisms were C/C in 6 ($12\%$) cases and C/T or T/T in 45 ($88\%$) cases. In the alcoholic patient group, polymorphisms were C/C in 12 ($26\%$) cases and C/T or T/T in 33 ($74\%$) cases (P=0.063) Conclusion: MTHFR polymorphisms are associated with gastric cancer and tumor site, but not with smoking and alcohol drinking.

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Studies on nutrient sources, fermentation and harmful organisms of the synthetic compost affecting yield of Agaricus bisporus (Lange) Sing (양송이 수량(收量)에 미치는 합성퇴비배지(合成堆肥培地)의 영양원(營養源), 발효(醱酵) 및 유해생물(有害生物)에 관((關)한 연구(硏究))

  • Shin, Gwan-Chull
    • The Korean Journal of Mycology
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    • v.7 no.1
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    • pp.13-73
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    • 1979
  • These studies were conducted to investigate nutrient sources and supplementary materials of synthetic compost media for Agaricus bisporus culture. Investigation were carried out to establish the optimum composition for compost of Agaricus bisporus methods of out-door fermentation and peakheating with rice straw as the main substrate of the media. The incidence and flora of harmful organisms in rice straw compost and their control were also studied. 1. When rice straw was used as the main substrate in synthetic compost as a carbon source. yields were remarkably high. Fermentation was more rapid than that of barley straw or wheat straw, and the total nitrogen content was high in rice straw compost. 2. Since the morphological and physico-chemical nature of Japonica and Indica types of rice straw are greatly dissimilar. there were apparent differences in the process of compost fermentation. Fermentation of Indica type straw proceeded more rapidly with a shortening the compost period, reducing the water supply, and required adding of supplementary materials for producing stable physical conditions. 3. Use of barley straw compost resulted in a smaller crop compared with rice straw. but when a 50%, barley straw and 50% rice straw mixture was used, the yield was almost the same as that using only rice straw. 4. There were extremely high positive correlations between yield of Agaricus bisporus and the total nitrogen, organic nitrogen, amino acids, amides and amino sugar nitrogen content of compost. The mycerial growth and fruit body formation were severely inhibited by ammonium nitrogen. 5. When rice straw was used as the main substrate for compost media, urea was the most suitable source of nitrogen. Poor results were obtained with calcium cyanamide and ammonium sulfate. When urea was applied three separate times, nitrogen loss during composting was decreased and the total nitrogen content of compost was increased. 6. The supplementation of organic nutrient activated compost fermentation and increased yield of Agaricus bisporus. The best sources of organic nutrients were: perilla meal, sesame meal, wheat bran and poultry manure, etc. 7. Soybean meal, tobacco powder and glutamic acid fermentation by-products which were industrial wastes, could be substituted for perilla meal, sesame meal and wheat bran as organic nutrient sources for compost media. B. When gypsum and zeolite were added to rice straw. physical deterioration of compost due to excess moisture and caramelization was observed. The Indica type of straw was more remarkable in increase of yield of Agricus bisporus by addition of supplementing materials than Japonica straw. 9. For preparing rice straw compost, the best mixture was prepared by 10% poultry manure, 5% perilla meal, 1. 2 to 1. 5% urea and 1% gypsum. At spring cropping, it was good to add rice bran to accelerate heat generation of the compost heap. 10. There was significantly high positive correlation (r=0.97) between accumulated temperature and the decomposition degree of compost during outdoor composting. The yield was highest at accumulated temperatures between 900 and $1,000^{\circ}C$. 11. Prolonging the composting period brought about an increase in decomposition degree and total nitrogen content, but a decrease in ammonium nitrogen. In the spring the suitable period of composting was 20 to 25 days. and about 15 days in autumn. For those periods, the degree of decomposition was 19 to 24%. 12. Compactness of wet compost at filling caused an increase in the residual ammonium nitrogen. methane and organic acid during peak heating. There was negative correlation between methane content and yield (r=0.76)and the same was true between volatile organic acid and yield (r=0.73). 13. In compost with a moisture content range between 69 to 80% at filling. the higher the moisture content, the lower the yield (r=0.78). This result was attributed to a reduction in the porosity of compost at filling the beds. The optimum porosity for good fermentation was between 41 and 53%. 14. Peak heating of the compost was essential for the prevention of harmful microorganisms and insect pests. and for the removal of excess ammonia. It was necessary to continue fer mentatiion for four days after peak heating. 15. Ten species of fungi which are harmful or competitive to Agaricus bisporus were identified from the rice compost, including Diehliomyces microsporus, Trichoderma sp. and Stysanus stemoites. The frequency of occurrance was notably high with serious damage to Agaricus bisporus. 16. Diehliomyces microsporus could be controlled by temperature adjustment of the growing room and by fumigating the compost and the house with Basamid and Vapam. Trichoderma was prevented by the use of Bavistin and Benomyl. 17. Four species of nematodes and five species of mites occured in compost during out-door composting. These orgnanisms could be controlled through peakheating compost for 6 hours at $60^{\circ}C$.

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