• Title/Summary/Keyword: M-1-1 Protocol

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Effects of Neurotrophic Factors on the Generation of Functional Dopamine Secretory Neurons Derived from in vitro Differentiated Human Embryonic Stem Cells (신경성장촉진 인자가 인간 배아줄기세포 유래 도파민 분비 신경세포형성에 미치는 영향)

  • Lee, Keum-Sil;Kim, Eun-Young;Shin, Hyun-Ah;Cho, Hwang-Yoon;Wang, Kyu-Chang;Kim, Yong-Sik;Lee, Hoon-Taek;Chung, Kil-Saeng;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.19-27
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    • 2004
  • Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

Effect of dietary mannanoligosaccharide supplementation on nutrient digestibility, hindgut fermentation, immune response and antioxidant indices in dogs

  • Pawar, Mahesh M.;Pattanaik, Ashok K.;Sinha, Dharmendra K.;Goswami, Tapas K.;Sharma, Kusumakar
    • Journal of Animal Science and Technology
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    • v.59 no.5
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    • pp.11.1-11.7
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    • 2017
  • Background: Use of prebiotics in companion animal nutrition is often considered advantageous over probiotics because of the ease of handling, ability to withstand processing and storage etc. While most of the studies on prebiotic use in dogs have been done with processed food as basal diet, the response in relation to homemade diet feeding is not very well explored. Methods: The study was conducted to evaluate the effects of dietary mannanoligosaccharide (MOS) supplementation on nutrient digestibility, hindgut fermentation, immune response and antioxidant indices in dogs. Ten Spitz pups were divided into two groups: control (CON) with no supplementation, and experimental (MOS) wherein the basal diet was supplemented with MOS at 15 g/kg diet. All dogs were fed on a home-prepared diet for a period of 150 days. The study protocol included a digestion trial, periodic blood collection and analysis for lipid profile and erythrocytic antioxidants. Immune response of the animals was assessed towards the end of the feeding period. Results: Results revealed no significant (P > 0.05) variations in palatability score, intake and apparent digestibility of nutrients between the groups. Faecal score, faeces voided, faecal pH, concentrations of ammonia, lactate and short-chain fatty acids were comparable (P > 0.05) between the two groups. Cell-mediated immune response, assessed as delayed-type of hypersensitivity response, was significantly higher (P < 0.05) in the MOS group. The percent of lymphocyte sub-populations CD4+ and ratio of CD4+:CD8+ were also significantly (P < 0.05) higher in MOS group. The serum IgG levels were similar (P > 0.05) in both the groups. Supplementation of MOS lowered (P < 0.05) serum total- and LDL- cholesterol levels, when compared with the control group. The erythrocytic antioxidant indices were similar (P > 0.05) between the two groups. Conclusions: The results indicated that supplementation of MOS at the rate of 15 g/kg in the diet of dog augmented the cell-mediated immune response and serum lipid profile without any influences on digestibility of nutrients, hindgut fermentation and antioxidants indices.

Vegetation Structure and Soil Properties of Ilex cornuta Population in Jeju Island (제주도 호랑가시나무개체군의 식생구조와 토양특성)

  • Kwon, Hye-Jin;Lee, Ji-Hye;Kim, Mu-Yeol;Lee, Jeong-Ho;Song, Ho-Kyung
    • Korean Journal of Environment and Ecology
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    • v.25 no.1
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    • pp.10-16
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    • 2011
  • This study was carried out to analyze the vegetation and soil characteristic, and ordination of the Ilex cornuta population, in the Yongsu Reservoir of Jeju Island. The Ilex cornuta population was classified into Cudrania tricuspidata dominant population, Sageretia theezans dominant population, and Mallotus japonicus dominant population. In the study sites, soil organic matter(O.M.), total nitrogen(T.N.), available phosphorus($P_2O_5$), changeable potassium, changeable calcium, changeable magnesium and soil pH were 14.62~17.35%, 0.39~0.51%, 8.83~20.15mg/kg, 0.44~0.64cmol+/kg, 5.79~6.87cmol+/kg, 3.43~4.19cmol+/kg and 5.41~5.80, respectively. The Mallotus japonicus dominant population was mainly found in the high percentage of available phosphorus, organic matter. It was also showed the lowest concentration of exchangeable K, and Mg. Cudrania tricuspidata dominant population and Sageretia theezans dominant population were found in the low percentage of available phosphorus, organic matter and there were also showed the highest concentration of exchangeable K, and Mg. It is one of the evidence that Ilex cornuta community indicated an intensive site management likes clearance of creeper is proper habitat management protocol. Thus, further researches should be followed to determine the other disturbance factors as management techniques.

Measurement of Microbial Protein Supply in Murrah Buffaloes (Bubalus bubalis) Using Urinary Purine Derivatives Excretion and PDC Index

  • Dipu, M.T.;George, S.K.;Singh, P.;Verma, A.K.;Mehra, U.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.347-355
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    • 2006
  • A study was conducted to predict the rumen microbial protein production based on urinary excretion of purine derivatives in buffaloes fed a diet of wheat straw and concentrate (40:60) at four fixed levels of feed intake. (95, 80, 60 and 40% of preliminary voluntary feed intake) following experimental protocol of IAEA (Phase I). The buffaloes were allocated according to a $4{\times}4$ latin square design. The urinary allantoin, uric acid, total PD excretion (mmol/d) in treatments L-95, L-80, L-60 and L-40 was 20.13, 16.00, 12.96 and 9.17; 1.88, 2.12, 2.11 and 2.15; 22.01, 18.12, 15.07 and 11.32, respectively and were significantly (p<0.05) different among treatments except for uric acid. The rate of PD excretion (mmol/d) was positively correlated with the digestible organic matter intake. Variations were observed in PD and creatinine concentration in spot samples collected at 6-hour interval. However, daily PD:Creatinine ratio (PDC index) appears to be a reasonably good predictor of microbial-N supply. The contribution of basal purine excretion to total excretion of purine derivatives (PD) was determined in pre-fasting period followed by a fasting period of 6 d (Phase II). Daily PD and creatinine excretion (mmol/kg $W^{0.75}$) during fasting averaged 0.117 and 0.456 respectively for buffaloes. The excretion rates of PD decreased significantly (p<0.01) during fasting compare to pre-fasting period, the urinary creatinine excretion remained almost similar. Except for creatinine, plasma concentration of target parameters significantly (p<0.01) declined during fasting. Likewise, glomerular filtration rate (GFR) and renal clearance of allantoin and uric acid also decreased. Based on the PD excretion rates during fasting and at different levels of feed intake obtained in this study, a relationship between daily urinary PD excretion (Y-mmol) and microbial purine absorption (X-mmol) was developed for buffaloes as Y = 0.74X+0.117 kg $W^{0.75}$. The microbial N supply (g/kg DOMI) remained statistically similar irrespective of dietary treatment. The results showed that excretion of urinary purine derivatives is positively correlated with the levels of feed intake in Murrah buffaloes and thus, estimation of urinary purine derivatives and PDC index could be used to determine microbial nitrogen supply when there is large variation in level of feed intake.

The Effects of Discrepancy in Reconstruction Algorithm between Patient Data and Normal Database in AutoQuant Evaluation: Focusing on Half-Time Scan Algorithm in Myocardial SPECT (심근 관류 스펙트에서 Half-Time Scan과 새로운 재구성법이 적용된 정상군 데이터를 기반으로 한 정량적 분석 결과의 차이 비교)

  • Lee, Hyung-Jin;Do, Yong-Ho;Cho, Seong-Wook;Kim, Jin-Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.122-126
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    • 2014
  • Purpose: The new reconstruction algorithms (NRA) provided by vendor aim to shorten the acquisition scan time. Whereas depending on the installed version AutoQuant program used for myocardial SPECT quantitative analysis did not contain the normal data that NRA is applied. Thus, the purpose of this paper is to compare the results according to AutoQuant versions in myocardial SPECT applied NRA and half-time scan (HT). Materials and Methods: Rest Tl and stress MIBI data of total 80 (40 men, 40 women) patients were gathered. Data were applied HT acquisition and ASTONISH (Philips) software which is NRA. Modified autoquant of SNUH and old version of AutoQuant (full-time scan) provided by company were compared. Comparison groups were classified as coronary artery disease (CAD), 24 hrs delay and almost normal patients who have a simple pain patient. Perfusion distribution aspect, summed stress score (SSS), summed rest score (SRS), extent and total perfusion deficit (TPD) of each 25 patient who have above diseases were compared and evaluated. Results: The case of CAD, when using re-edited AutoQuant (HT) SSS and SRS showed about 30% reduction (P<0.0001), Extent showed about 38% reduction and TPD showed about 30% reduction in the tendency (P<0.0001). In the score of the perfusion, especially on the part of infero-medium, infero-apical, lateral-medium and lateral-apical regions were the biggest change. The case of the 24 hrs delay patient SRS (P=0.042), Extent (P=0.018) and TPD (P=0.0024) showed about 13-18% reduction. And the case of simple pain patient, comparison of 4 results showed about 5-7% reduction. Conclusion: This study was started based on expectation that results could be affected by normal patient data. Normal patient data is possible to change by race and gender. It was proved that combination of new reconstruction algorithm for reducing scan time and analysis program according to scan protocol with NRA could also be affected to results. Clinical usefulness of gated myocardial SPECT is possibly increased if each hospital properly collects normal patient data for their scan acquisition protocol.

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Dosimetry of the Low Fluence Fast Neutron Beams for Boron Neutron Capture Therapy (붕소-중성자 포획치료를 위한 미세 속중성자 선량 특성 연구)

  • Lee, Dong-Han;Ji, Young-Hoon;Lee, Dong-Hoon;Park, Hyun-Joo;Lee, Suk;Lee, Kyung-Hoo;Suh, So-Heigh;Kim, Mi-Sook;Cho, Chul-Koo;Yoo, Seong-Yul;Yu, Hyung-Jun;Gwak, Ho-Shin;Rhee, Chang-Hun
    • Radiation Oncology Journal
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    • v.19 no.1
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    • pp.66-73
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    • 2001
  • Purpose : For the research of Boron Neutron Capture Therapy (BNCT), fast neutrons generated from the MC-50 cyclotron with maximum energy of 34.4 MeV in Korea Cancer Center Hospital were moderated by 70 cm paraffin and then the dose characteristics were investigated. Using these results, we hope to establish the protocol about dose measurement of epi-thermal neutron, to make a basis of dose characteristic of epi-thermal neutron emitted from nuclear reactor, and to find feasibility about accelerator-based BNCT. Method and Materials : For measuring the absorbed dose and dose distribution of fast neutron beams, we used Unidos 10005 (PTW, Germany) electrometer and IC-17 (Far West, USA), IC-18, ElC-1 ion chambers manufactured by A-150 plastic and used IC-l7M ion chamber manufactured by magnesium for gamma dose. There chambers were flushed with tissue equivalent gas and argon gas and then the flow rate was S co per minute. Using Monte Carlo N-Particle (MCNP) code, transport program in mixed field with neutron, photon, electron, two dimensional dose and energy fluence distribution was calculated and there results were compared with measured results. Results : The absorbed dose of fast neutron beams was $6.47\times10^{-3}$ cGy per 1 MU at the 4 cm depth of the water phantom, which is assumed to be effective depth for BNCT. The magnitude of gamma contamination intermingled with fast neutron beams was $65.2{\pm}0.9\%$ at the same depth. In the dose distribution according to the depth of water, the neutron dose decreased linearly and the gamma dose decreased exponentially as the depth was deepened. The factor expressed energy level, $D_{20}/D_{10}$, of the total dose was 0.718. Conclusion : Through the direct measurement using the two ion chambers, which is made different wall materials, and computer calculation of isodose distribution using MCNP simulation method, we have found the dose characteristics of low fluence fast neutron beams. If the power supply and the target material, which generate high voltage and current, will be developed and gamma contamination was reduced by lead or bismuth, we think, it may be possible to accelerator-based BNCT.

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The Extended Site Assessment Procedure Based on Knowledge of Biodegradability to Evaluate the Applicability of Intrinsic Remediation (자연내재복원기술(Intrinsic Remediation)적용을 위한 오염지역 평가과정 개발)

  • ;Robert M. Cowan
    • Journal of Korea Soil Environment Society
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    • v.2 no.3
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    • pp.3-21
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    • 1997
  • The remediation of contamiated sites using currently available remediation technologies requires long term treatment and huge costs, and it is uncertain to achieve the remediation goal to drop contamination level to either back-ground or health-based standards by using such technologies. Intrinsic remediation technology is the remediation technology that relies on the mechanisms of natural attenuation for the containment and elimination of contaminants in subsurface environments. Initial costs for the intrinsic remediation may be higher than conventional treatment technologies because the most comprehensive site assessment for intrinsic remediation is required. Total remediation cost, however may be the lowest among the presently employed technologies. The applicability of intrinsic remediation in the contaminated sites should be theroughly investigated to achieve the remedial goal of the technology. This paper provides the frame of the extended site assessment procedure based on knowledge of biodegradability to evaluate the applicability of intrinsic remediation. This site assessment procedure is composed of 5 steps such as preliminary site screening, assessment of the current knowledge of biodegradability, selecting the appropriate approach, analyzing the contaminant fate and transport and planning the monitoring schedule. In the step 1, followings are to be decided 1) whether to go on the the detailed assessment or not based on the rules of thumb concerning the biodegradability of organic compounds, 2) which protocol document is selected to follow for detailed site assessment according to the site characteristics, contaminants and the relative distance between the contamination and potential receptors. In the step 2, the database for biodegradability are searched and evaluated. In the step 3, the appropriate biodegradability pathways for the contaminated site is selected. In the step 4, the fate and transport of the contaminants at the site are analyzed through modeling. In the step 5, the monitoring schedule is planned according to the result of the modeling. Through this procedure, users may able to have the rational and systematic informations for the application of intrinsic remediation. Also the collected data and informations can be used as the basic to re-select the other remediation technology if it reaches a conclusion not to applicate intrinsic remediation technology at the site from the site assessment procedure.

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Corrections on CH4 Fluxes Measured in a Rice Paddy by Eddy Covariance Method with an Open-path Wavelength Modulation Spectroscopy (개회로 파장 변조 분광법과 에디 공분산 방법으로 논에서 관측된 CH4 플럭스 자료의 보정)

  • Kang, Namgoo;Yun, Juyeol;Talucder, M.S.A.;Moon, Minkyu;Kang, Minseok;Shim, Kyo-Moon;Kim, Joon
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.17 no.1
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    • pp.15-24
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    • 2015
  • $CH_4$ is a trace gas and one of the key greenhouse gases, which requires continuous and systematic monitoring. The application of eddy covariance technique for $CH_4$ flux measurement requires a fast-response, laser-based spectroscopy. The eddy covariance measurements have been used to monitor $CO_2$ fluxes and their data processing procedures have been standardized and well documented. However, such processes for $CH_4$ fluxes are still lacking. In this note, we report the first measurement of $CH_4$ flux in a rice paddy by employing the eddy covariance technique with a recently commercialized wavelength modulation spectroscopy. $CH_4$ fluxes were measured for five consecutive days before and after the rice transplanting at the Gimje flux monitoring site in 2012. The commercially available $EddyPro^{TM}$ program was used to process these data, following the KoFlux protocol for data-processing. In this process, we quantified and documented the effects of three key corrections: (1) frequency response correction, (2) air density correction, and (3) spectroscopic correction. The effects of these corrections were different between daytime and nighttime, and their magnitudes were greater with larger $CH_4$ fluxes. Overall, the magnitude of $CH_4$ flux increased on average by 20-25% after the corrections. The National Center for AgroMeteorology (www.ncam.kr) will soon release an updated KoFlux program to public users, which includes the spectroscopic correction and the gap-filling of $CH_4$ flux.

Analysis of Telomere Length and Telomerase Activity of Tissues in Korean Native Chicken (한국 재래닭의 주령별 각 조직의 텔로미어 함량과 텔로머레이스 활성도 분석)

  • Jung G.S.;Cho E.J.;Choi D.S.;Lee M.J.;Park C.;Jeon I.S.;Sohn S.H.
    • Korean Journal of Poultry Science
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    • v.33 no.2
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    • pp.97-103
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    • 2006
  • Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Even though telomere length and telomerase activity have been studied extensively, very little is known to analyze the telomere dynamics in chicken cells. This study was carried out to analyze the telomere distribution and telomerase activity of Korean Native Chicken cells along with aging. The cells were collected from brain, heart, liver, kidney and germinal tissues during physiological stages. Telomere distribution was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was performed by Telomeric Repeat Amplification Protocol (TRAP) assay. In results, the telomeres of chicken were found at the ends of all chromosomes with the interstitial telomeres on chromosomes 1, 2 and 3. The amount of telomeres on chicken cells was decreased along with aging in most tissues. Furthermore, the telomere quantity was significantly different among tissues. The relative amount of telomeres in proliferous cells such as testis cells had much more than those of liver, brain, heart, blood and kidney cells. The telomerase activity was down-regulated in cells of brain, heart and liver tissues. Whereas gonadal cells showed a constitutive activity of telomerase during all stage of life. In conclusions, the telomere quantity and telomerase activity in chicken are closely relate to cell proliferation and tissue specificity during developmental stages and aging. There is also closely correlated between the amounts of telomeric DNA and telomerase activity in chicken tissues.