• Title/Summary/Keyword: Development Cycle

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Ultrasensitive Enzymeimmunoassay for Testosterone in Human Saliva (사람 타액내 Testosterone의 초감도 효소면역측정법)

  • 윤용달;전은현;이창주;도병록;이준영
    • Development and Reproduction
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    • v.4 no.1
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    • pp.115-123
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    • 2000
  • A few enzymeimmunoassay (EfA) for testosterone (T) have been reported but was not suitable for all biological samples. The present study was designed to develop a rapid, ultrasensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected at 06:00~09:00 hour during one menstrual cycle from 18 normally menstruating women and on 09:00~10:00 hour from 20 normal men. The present study shows an established EIA for testosterone, using horseradish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxymethyloxime (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by a mixed anhydride method in order to facilitate separation of bound and free steroids. The established EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay. The sensitivity of the established EIA was less than 25 pg/tube. The correlation coefficients between the expected T-values and observed T-values measured by EIA or RIA were r=0.985 and r=0.941 respectively. The cross reactivity of antiserum in EIA was a little higher than that of RIA, especially by 5 ${\alpha}$-DHT. The intra- and inter-assay precisions of the present EIA were similar to those of RIA. The present study also demonstrates that the normal T-values in saliva of Korean male & female samples are 265.65${\pm}$15.80 pmol/l and 109.74${\pm}$ 12.01 pmol/l, respectively. The present EIA seems to be established and suitable for use in the endocrinological studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine assay of ordinary laboratory with a conventional spectrophotometer.

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Caspase-3 Activation is Associated with Granulosa Cell Apoptosis during Follicular Atresia in Porcine Ovary (돼지 폐쇄난포내 과립세포의 자연세포사 시 캐스파제-3의 활성화)

  • Kim, Jong-Min;Chung, Jin-Yong;Kim, Ji-Young;Oh, Seung-Hoon;Song, Kang-Won;Do, Byoung-Rok;Kim, Sang-Soo;Jung, Jin;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.10 no.1
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    • pp.1-7
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    • 2006
  • Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.

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Anti-adipogenic Activity of Cortex ulmi pumilae Extract in 3T3-L1 Preadipocytes (유근피 추출물의 3T3-L1지방전구세포의 분화 억제 효능에 관한 연구)

  • Jeong, Hyun Young;Jin, Soojung;Nam, Soo Wan;Hyun, Sook Kyung;Kim, Sung Gu;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.24 no.2
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    • pp.137-147
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    • 2014
  • Cortex ulmi pumilae, the cortex of Ulmus davidiana var. japonica, has been used in traditional folk medicine for its anti-inflammatory effect. Although its various bioactivities such as anti-inflammatory, anti-microbial, and anti-cancer, have been reported, the anti-adipogenic activity of cortex ulmi pumilae remains unclarified. In the present study, we investigated the effect of cortex ulmi pumilae extract on adipocyte differentiation in 3T3-L1 preadipocytes. Treatment with cortex ulmi pumilae extract significantly reduced the formation of lipid droplets and triglyceride content in a dose-dependent manner; this is associated with an inhibition of the adipogenic transcription factors, CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$). In addition, cortex ulmi pumilae extract treatment during the early stage of adipogenesis showed more efficient anti-adipogenic activity than treatment during other stages of adipogenesis. Cortex ulmi pumilae extract also inhibited cell proliferation and induced G1 arrest of 3T3-L1 cells in the early stage of adipogenesis. This was associated with upregulated expression of Cdk inhibitor p21 and downregulated expression of cyclin E and phospho-Rb, indicating that cortex ulmi pumilae extract blocks mitotic clonal expansion by cell cycle regulation. Taken together, these results suggest that cortex ulmi pumilae extract possesses anti-adipogenic activity through the inhibition of adipocyte differentiation by blocking mitotic clonal expansion.

Gene Expression Profiling of SH-SY5Y Human Neuroblastoma Cells Treated with Ginsenoside Rg1 and Rb1 (Ginsenoside Rg1 및 Rb1을 처리한 신경세포주(SH-SY5Y세포)의 유전자 발현양상)

  • Lee, Joon-Noh;Yang, Byung-Hwan;Choi, Seung-Hak;Kim, Seok-Hyun;Chai, Young-Gyu;Jung, Kyoung-Hwa;Lee, Jun-Seok;Choi, Kang-Ju;Kim, Young-Suk
    • Korean Journal of Biological Psychiatry
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    • v.12 no.1
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    • pp.42-61
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    • 2005
  • Objectives:The ginsenoside Rg1 and Rb1, the major components of ginseng saponin, have neurotrophic and neuroprotective effects including promotion of neuronal survival and proliferation, facilitation of learning and memory, and protection from ischemic injury and apoptosis. In this study, to investigate the molecular basis of the effects of ginsenoside on neuron, we analyzed gene expression profiling of SH-SY5Y human neuroblastoma cells treated with ginsenoside Rg1 or Rb1. Methods:SH-SY5Y cells were cultured and treated in triplicate with ginsenoside Rg1 or Rb1($80{\mu}M$, $40{\mu}M$, $20{\mu}M$). The proliferation rates of SH-SY5Y cells were determined by MTT assay and microscopic examination. We used a high density cDNA microarray chip that contained 8K human genes to analyze the gene expression profiles in SH-SY5Y cells. We analyzed using the Significance Analysis of Microarray(SAM) method for identifying genes on a microarray with statistically significant changes in expression. Results:Treatment of SH-SY5Y cells with $80{\mu}M$ ginsenoside Rg1 or Rb1 for 36h showed maximal proliferation compared with other concentrations or control. The results of the microarray experiment yielded 96 genes were upregulated(${\geq}$3 fold) in Rg1 treated cells and 40 genes were up-regulated(${\geq}$2 fold) in Rb1 treated cells. Treatment with ginsenoside Rg1 for 36h induced the expression of some genes associated with protein biosynthesis, regulation of transcription or translation, cell proliferation and growth, neurogenesis and differentiation, regulation of cell cycle, energy transport and others. Genes associated with neurogenesis and neuronal differentiation such as SCG10 and MLP increased in ginsenoside Rg1 treated cells, but such changes did not occur in Rb1-group. Conclusion:Our data provide novel insights into the gene mechanisms involved in possible role for ginsenoside Rg1 or Rb1 in mediating neuronal proliferation or cell viability, which can elicit distinct patterns of gene expression in neuronal cell line. Ginsenoside Rg1 have more broad and strong effects than ginsenoside Rb1 in gene expression and related cellular physiology. In addition, we suggest that SCG10 gene, which is known to be expressed in neuronal differentiation during development and neuronal regeneration during adulthood, may have a role in enhancement of activity dependent synaptic plasticity or cytoskeletal regulation following treatment of ginsenoside Rg1. Further, ginsenoside Rg1 may have a possible role in regeneration of injured neuron, promotion of memory, and prevention from aging or neuronal degeneration.

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The Influential Factor Analysis in the Technology Valuation of The Agri-Food Industry and the Simulation-Based Valuation Analysis (농식품 산업의 기술평가 영향요인 분석과 시뮬레이션 기반 기술평가 비교)

  • Kim, Sang-gook;Jun, Seung-pyo;Park, Hyun-woo
    • Journal of Technology Innovation
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    • v.24 no.4
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    • pp.277-307
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    • 2016
  • Since 2011, DCF(Discounted Cash Flow) method has been used initiatively for valuating R&D technology assets in the agricultural food industry and recently technology valuation based on royalties comparison among technology transfer transactions has been also carried out in parallel when evaluating the technology assets such as new seed development technologies. Since the DCF method which has been known until now has many input variables to be estimated, sophisticated estimation has been demanded at the time of technology valuation. In addition, considering more similar trading cases when applying sales transaction comparison or industry norm method based on information of technology transfer royalty, it is an important issue that should be taken into account in the same way in the Agri-Food industry. The main input variables used for technology valuation in the Agri-Food industry are life cycle of technology asset, the financial information related to the Agri-Food industry, discount rate, and technology contribution rate. The latest infrastructure building and data updating related to technology valuation has been carried out on a regular basis in the evaluation organization of the Agri-Food segment. This study verifies the key variables that give the most important impact on the results for the existing technology valuation in the Agri-Food industry and clarifies the difference between the existing valuation result and the outcome by referring the support information that is derived through the latest input information applied in DCF method. In addition, while presenting the scheme to complement fragment information which the latest input data just influence result of technology valuation, we tried to perform comparative analysis between the existing valuation results and the evaluated outcome after the latest of reference data for making a decision the input values to be estimated in DCF. To perform these analyzes, it was first selected the representative cases evaluated past in the Agri-Food industry, applied a sensitivity analysis for input variables based on these selected cases, and then executed a simulation analysis utilizing the key input variables derived from sensitivity analysis. The results of this study is to provide the information which there are the need for modernization of the data related to the input variables that are utilized during valuating technology assets in the Agri-Food sector and for building the infrastructure of the key input variables in DCF. Therefore it is expected to provide more fruitful information about the results of valuation.

Alternative Concept to Enhance the Disposal Efficiency for CANDU Spent Fuel Disposal System (CANDU 사용후핵연료 처분시스템 효율향상 개념 도출)

  • Lee, Jong-Youl;Cho, Dong-Geun;Kook, Dong-Hak;Lee, Min-Soo;Choi, Heui-Joo
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.9 no.3
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    • pp.169-179
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    • 2011
  • There are two types of nuclear reactors in Korea and they are PWR type and CANDU type. The safe management of the spent fuels from these reactors is very important factor to maintain the sustainable energy supply with nuclear power plant. In Korea, a reference disposal system for the spent fuels has been developed through a study on the direct disposal of the PWR and CANDU spent fuel. Recently, the research on the demonstration and the efficiency analyses of the disposal system has been performed to make the disposal system safer and more economic. PWR spent fuels which include a lot of reusable material can be considered being recycled and a study on the disposal of HLW from this recycling process is being performed. CANDU spent fuels are considered being disposed of directly in deep geological formation, since they have little reusable material. In this study, based on the Korean Reference spent fuel disposal System (KRS) which was to dispose of both PWR type and CANDU type, the more effective CANDU spent fuel disposal systems were developed. To do this, the disposal canister for CANDU spent fuels was modified to hold the storage basket for 60 bundles which is used in nuclear power plant. With these modified disposal canister concepts, the disposal concepts to meet the thermal requirement that the temperature of the buffer materials should not be over $100^{\circ}C$ were developed. These disposal concepts were reviewed and analyzed in terms of disposal effective factors which were thermal effectiveness, U-density, disposal area, excavation volume, material volume etc. and the most effective concept was proposed. The results of this study will be used in the development of various wastes disposal system together with the HLW wastes from the PWR spent fuel recycling process.

Study on Basic Requirements of Geoscientific Area for the Deep Geological Repository of Spent Nuclear Fuel in Korea (사용후핵연료 심지층처분장부지 지질환경 기본요건 검토)

  • Bae, Dae-Seok;Koh, Yong-Kwon;Park, Ju-Wan;Park, Jin-Baek;Song, Jong-Soon
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.10 no.1
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    • pp.63-75
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    • 2012
  • This paper gives some basic requirements and preferences of various geological environmental conditions for the final deep geological repository of spent nuclear fuel (SNF). This study also indicates how the requirements and preferences are to be considered prior to the selection of sites for a site investigation as well as the final disposal in Korea. The results of the study are based on the knowledge and experience from the IAEA and NEA/OECD as well as the advanced countries in SNF disposal project. This study discusses and suggests preliminary guideline of the disposal requirements including geological, mechanical, thermal, hydrogeological, chemical and transport properties of host rock with long term geological stabilities which influence the functions of a multi-barrier disposal system. To apply and determine whether requirements and preferences for a given parameter are satisfied at different stages during a site selection and suitability assessment of a final disposal site, the quantitative criteria in each area should be formulated with credibility through relevant research and development efforts for the deep geological environment during the site screening and selection processes as well as specific studies such as productions of safety cases and validation studies using a generic underground research laboratory (URL) in Korea.

Efficacy of Frozen-Thawed ET in Patients with Old Age or Non-Pregnant in Fresh ET Cycles (고령 환자와 신선주기 배아이식에서 임신에 실패한 환자에서 동결-융해 배아이식의 효용성)

  • Choi, Su Jin;Lee, Sun Hee;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.237-243
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    • 2006
  • Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

mRNA Expression Differences of uPA, uPAR in Eutopic Endometrium of Advanced Stage Endometriosis Patients (중증 자궁내막증 환자의 자궁내막과 정상인 자궁내막에서 uPA, uPAR mRNA 발현의 차이에 관한 연구)

  • Hur, Sung Eun;Lee, Ji Young;Lee, Woon Jung;Chung, Hye-Sung;Chung, Hye Won
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.229-236
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    • 2006
  • Objective: We investigated the expression of uPA and uPAR in eutopic endometrium of advanced stage endometriosis and control patients. Methods: The 33 endometriosis patients and 32 controls were enrolled. Endometrial samples were obtained from 65 premenopausal women aged 29-44 years, undergoing laparoscopic surgery or hysterectomy for non-malignant lesions. Sufficient samples were collected from 33 patients with endometriosis stage Ill and IV and 32 controls without endometriosis confirmed by laparoscopic surgery. The mRNA expression of uPA and uPAR from eutopic endometrium were analyzed by RT-QC PCR. Results: The mRNAs of uPA and uPAR were expressed in eutopic endometrium from endometriosis and normal controls throughout the menstrual cycle. Uterine endometrium from women with endometriosis expresses significantly (p<0.05) higher levels of u-PA mRNA than endometrium from normal women without endometriosis in the proliferative phase. There were no significant differences in expression of uPAR in eutopic endometrium between controls and endometriosis patients. Conclusion: These results suggest that eutopic endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation because of greater u-PA mRNA expression than endometrium from women without endometriosis. Thus, increased proteolytic activity may be one etiology for the invasive properties of the endometrium resulting in the development of endometriosis.

Structural Behavior of Mixed $LiMn_2O_4-LiNi_{1/3}Co_{1/3}Mn_{1/3}O_2$ Cathode in Li-ion Cells during Electrochemical Cycling

  • Yun, Won-Seop;Lee, Sang-U
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2011.05a
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    • pp.5-5
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    • 2011
  • The research and development of hybrid electric vehicle (HEV), plug-in hybrid electric vehicle (PHEV) and electric vehicle (EV) are intensified due to the energy crisis and environmental concerns. In order to meet the challenging requirements of powering HEV, PHEV and EV, the current lithium battery technology needs to be significantly improved in terms of the cost, safety, power and energy density, as well as the calendar and cycle life. One new technology being developed is the utilization of composite cathode by mixing two different types of insertion compounds [e.g., spinel $LiMn_2O_4$ and layered $LiMO_2$ (M=Ni, Co, and Mn)]. Recently, some studies on mixing two different types of cathode materials to make a composite cathode have been reported, which were aimed at reducing cost and improving self-discharge. Numata et al. reported that when stored in a sealed can together with electrolyte at $80^{\circ}C$ for 10 days, the concentrations of both HF and $Mn^{2+}$ were lower in the can containing $LiMn_2O_4$ blended with $LiNi_{0.8}Co_{0.2}O_2$ than that containing $LiMn_2O_4$ only. That reports clearly showed that this blending technique can prevent the decline in capacity caused by cycling or storage at elevated temperatures. However, not much work has been reported on the charge-discharge characteristics and related structural phase transitions for these composite cathodes. In this presentation, we will report our in situ x-ray diffraction studies on this mixed composite cathode material during charge-discharge cycling. The mixed cathodes were incorporated into in situ XRD cells with a Li foil anode, a Celgard separator, and a 1M $LiPF_6$ electrolyte in a 1 : 1 EC : DMC solvent (LP 30 from EM Industries, Inc.). For in situ XRD cell, Mylar windows were used as has been described in detail elsewhere. All of these in situ XRD spectra were collected on beam line X18A at National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory using two different detectors. One is a conventional scintillation detector with data collection at 0.02 degree in two theta angle for each step. The other is a wide angle position sensitive detector (PSD). The wavelengths used were 1.1950 ${\AA}$ for the scintillation detector and 0.9999 A for the PSD. The newly installed PSD at beam line X18A of NSLS can collect XRD patterns as short as a few minutes covering $90^{\circ}$ of two theta angles simultaneously with good signal to noise ratio. It significantly reduced the data collection time for each scan, giving us a great advantage in studying the phase transition in real time. The two theta angles of all the XRD spectra presented in this paper have been recalculated and converted to corresponding angles for ${\lambda}=1.54\;{\AA}$, which is the wavelength of conventional x-ray tube source with Cu-$k{\alpha}$ radiation, for easy comparison with data in other literatures. The structural changes of the composite cathode made by mixing spinel $LiMn_2O_4$ and layered $Li-Ni_{1/3}Co_{1/3}Mn_{1/3}O_2$ in 1 : 1 wt% in both Li-half and Li-ion cells during charge/discharge are studied by in situ XRD. During the first charge up to ~5.2 V vs. $Li/Li^+$, the in situ XRD spectra for the composite cathode in the Li-half cell track the structural changes of each component. At the early stage of charge, the lithium extraction takes place in the $LiNi_{1/3}Co_{1/3}Mn_{1/3}O_2$ component only. When the cell voltage reaches at ~4.0 V vs. $Li/Li^+$, lithium extraction from the spinel $LiMn_2O_4$ component starts and becomes the major contributor for the cell capacity due to the higher rate capability of $LiMn_2O_4$. When the voltage passed 4.3 V, the major structural changes are from the $LiNi_{1/3}Co_{1/3}Mn_{1/3}O_2$ component, while the $LiMn_2O_4$ component is almost unchanged. In the Li-ion cell using a MCMB anode and a composite cathode cycled between 2.5 V and 4.2 V, the structural changes are dominated by the spinel $LiMn_2O_4$ component, with much less changes in the layered $LiNi_{1/3}Co_{1/3}Mn_{1/3}O_2$ component, comparing with the Li-half cell results. These results give us valuable information about the structural changes relating to the contributions of each individual component to the cell capacity at certain charge/discharge state, which are helpful in designing and optimizing the composite cathode using spinel- and layered-type materials for Li-ion battery research. More detailed discussion will be presented at the meeting.

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