• Title/Summary/Keyword: time-dependent effect

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Effects of Endocrine Disruptors (NP, DBP and BPA) on Sperm Characteristics and Development of IVF Embryos in Pig

  • Yuh, In Suh;Cheong, Hee Tae;Kim, Jong Taek;Park, In Chul;Park, Choon Keun;Yang, Boo Keun
    • Journal of Animal Science and Technology
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    • v.55 no.4
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    • pp.237-247
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    • 2013
  • This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, $30{\mu}M$), DBP (10, 50, $100{\mu}M$) and BPA (1, 5 or $10{\mu}g/ml$) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time. In combination effects, NP ($10{\mu}M$) + DBP ($10{\mu}M$) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; $30{\mu}M$, DBP; $100{\mu}M$) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$) on sperm characteristics were larger than those of NP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) (p<0.05). DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) decreased sperm characteristics compared with the low concentration combination (DBP $10{\mu}M$ + BPA $1{\mu}g/ml$, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP ($30{\mu}M$) + DBP ($100{\mu}M$), NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$), DBP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) or DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control. NP (${\geq}20{\mu}M$) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or $10{\mu}g/ml$ decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP ($100{\mu}M$) plus NP ($30{\mu}M$) and DBP ($100{\mu}M$) plus BPA ($10{\mu}g/ml$) did not affect on morulae and blastocyst development, but NP ($30{\mu}M$) plus BPA ($10{\mu}g/ml$) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.

Effect of Extraction Conditions on in vitro Antioxidant Activities of Root Bark Extract from Ulmus pumila L. (추출조건에 따른 유근피 추출물의 항산화 활성)

  • Kim, Jae-Min;Cho, Myoung-Lae;Seo, Kyu-Eun;Kim, Ye-Seul;Jung, Tae-Dong;Kim, Young-Hyun;Kim, Dan-Bi;Shin, Gi-Hae;Oh, Ji-Won;Lee, Jong Seok;Lee, Jin-Ha;Kim, Jong-Yae;Lee, Dae-Won;Lee, Ok-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1172-1179
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    • 2015
  • This study investigated optimal extraction conditions for application of Ulmus pumila L. as a natural antioxidant. U. pumila L. was extracted using ethanol (EtOH) at various concentrations (0, 40, and 80%) and extraction times (1, 2, and 3 h) at $70^{\circ}C$ and then evaluated for extraction yield, total phenolic contents, total flavonoid contents, as well as antioxidant activities [2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, reducing power, and oxygen radical absorbing capacity (ORAC)]. Antioxidant activities were correlated with total phenolic and flavonoid contents. Of the solvent conditions, 80% EtOH extracts for 3 h at $70^{\circ}C$ showed the highest total phenolic and flavonoid contents with strong antioxidant activities, although there were no significant time effects on DPPH and ABTS radical scavenging activities and reducing power. However, ORAC values of all EtOH extracts remarkably increased in a time-dependent manner. In addition, 80% EtOH extract for 3 h exhibited strong antioxidant effects on HDF and 3T3-L1 cells. Therefore, the antioxidant capacity of U. pumila L., may due to phenolic and flavonoid contents, and extraction conditions were 80% EtOH for 3 h at $70^{\circ}C$. This extract could be a good source for natural antioxidants.

A Regression-Model-based Method for Combining Interestingness Measures of Association Rule Mining (연관상품 추천을 위한 회귀분석모형 기반 연관 규칙 척도 결합기법)

  • Lee, Dongwon
    • Journal of Intelligence and Information Systems
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    • v.23 no.1
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    • pp.127-141
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    • 2017
  • Advances in Internet technologies and the proliferation of mobile devices enabled consumers to approach a wide range of goods and services, while causing an adverse effect that they have hard time reaching their congenial items even if they devote much time to searching for them. Accordingly, businesses are using the recommender systems to provide tools for consumers to find the desired items more easily. Association Rule Mining (ARM) technology is advantageous to recommender systems in that ARM provides intuitive form of a rule with interestingness measures (support, confidence, and lift) describing the relationship between items. Given an item, its relevant items can be distinguished with the help of the measures that show the strength of relationship between items. Based on the strength, the most pertinent items can be chosen among other items and exposed to a given item's web page. However, the diversity of the measures may confuse which items are more recommendable. Given two rules, for example, one rule's support and confidence may not be concurrently superior to the other rule's. Such discrepancy of the measures in distinguishing one rule's superiority from other rules may cause difficulty in selecting proper items for recommendation. In addition, in an online environment where a web page or mobile screen can provide a limited number of recommendations that attract consumer interest, the prudent selection of items to be included in the list of recommendations is very important. The exposure of items of little interest may lead consumers to ignore the recommendations. Then, such consumers will possibly not pay attention to other forms of marketing activities. Therefore, the measures should be aligned with the probability of consumer's acceptance of recommendations. For this reason, this study proposes a model-based approach to combine those measures into one unified measure that can consistently determine the ranking of recommended items. A regression model was designed to describe how well the measures (independent variables; i.e., support, confidence, and lift) explain consumer's acceptance of recommendations (dependent variables, hit rate of recommended items). The model is intuitive to understand and easy to use in that the equation consists of the commonly used measures for ARM and can be used in the estimation of hit rates. The experiment using transaction data from one of the Korea's largest online shopping malls was conducted to show that the proposed model can improve the hit rates of recommendations. From the top of the list to 13th place, recommended items in the higher rakings from the proposed model show the higher hit rates than those from the competitive model's. The result shows that the proposed model's performance is superior to the competitive model's in online recommendation environment. In a web page, consumers are provided around ten recommendations with which the proposed model outperforms. Moreover, a mobile device cannot expose many items simultaneously due to its limited screen size. Therefore, the result shows that the newly devised recommendation technique is suitable for the mobile recommender systems. While this study has been conducted to cover the cross-selling in online shopping malls that handle merchandise, the proposed method can be expected to be applied in various situations under which association rules apply. For example, this model can be applied to medical diagnostic systems that predict candidate diseases from a patient's symptoms. To increase the efficiency of the model, additional variables will need to be considered for the elaboration of the model in future studies. For example, price can be a good candidate for an explanatory variable because it has a major impact on consumer purchase decisions. If the prices of recommended items are much higher than the items in which a consumer is interested, the consumer may hesitate to accept the recommendations.

The Effect of Epigallocatechin-3-gallate on HIF-1 α and VEGF in Human Lung Cancer Cell Line (비소세포폐암주에서 저산소상태에 의해 유발된 HIFa-1 α와 VEGF의 발현증가에 미치는 Epigallocatechin-3-gallate의 억제 효과)

  • Song, Joo Han;Jeon, Eun Joo;Kwak, Hee Won;Lee, Hye Min;Cho, Sung Gun;Kang, Hyung Koo;Park, Sung Woon;Lee, Jae Hee;Lee, Byung Ook;Jung, Jae Woo;Choi, Jae Cheol;Shin, Jong Wook;Kim, Ki Jeong;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.3
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    • pp.178-185
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    • 2009
  • Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

Role of p-38 MAP Kinase in apoptosis of hypoxia-induced osteoblasts (저산소 상태로 인한 조골세포 고사사기전에서 p-38 MAP kinase의 역할에 관한 연구)

  • Yoon, Jeong-Hyeon;Jeong, Ae-Jin;Kang, Kyung-Hwa;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.33 no.3 s.98
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    • pp.169-183
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    • 2003
  • Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.

Association of Genetic Variations with Pemetrexed-Induced Cytotoxicity in Non-Small Cell Lung Cancer Cells (비소세포폐암 세포주에서 pemetrexed의 세포독성과 유전학적 다형성과의 상관성 조사)

  • Yoon, Seong-Ae;Choi, Jung-Ran;Kim, Jeong-Oh;Shin, Jung-Young;Zhang, XiangHua;Kang, Jin-Hyoung
    • Journal of Life Science
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    • v.20 no.1
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    • pp.103-112
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    • 2010
  • Pemetrexed has demonstrated clinical activity in non-small cell lung cancer (NSCLC) as well as other solid tumors. It transports into the cells via reduced folate carrier (RFC) and is polyglutamated by folypolyglutamate synthetase (FPGS). Pemetrexed directly inhibits several folate-dependent enzymes such as thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). We investigated the effects of genetic variations and the expression of RFC, FPGS, TS and DHFR enzymes on drug sensitivity to pemetrexed in NSCLC cells. Polymorphisms in RFC, FPGS, and DHFR were genotyped in four NSCLC cells - A549, PC14, HCC-1588, and H226. Real-time RT-PCR and Western blot was performed to evaluate mRNA transcripts and protein of these genes. The cytotoxicity of pemetrexed was measured by SRB assay. In PC14 and H226 cells, increased mRNA expressions of RFC and FPGS were associated with higher cytotoxicity to pemetrexed. 2R/2R genotype of TS and its increased mRNA expression were associated with drug resistance to pemetrexed in A549 cells, whereas 3R/3R genotype in TS with decreased mRNA expression was associated with higher sensitivity in H226 cells. After pemetrexed treatment, an inverse change of DHFR mRNA and protein expression was found. The strongest linkage disequilibrium (LD) was discovered between-1726C>T and -1188A>C SNP of DHFR gene. Our findings suggest the cytotoxic effect of pemetrexed may be associated with genetic polymorphisms and the expression level of genes involved in pemetrexed metabolisms in NSCLC cells.

Effect of Temperature on Development and Life Table Parameters of Tetranychus urticae Koch (Acari: Tetranychide) Reared on Eggplants (가지에서 온도별 점박이응애 발육특성 및 생명표 통계량)

  • Kim, Ju;Lee, Sang-Koo;Kim, Jeong-Man;Kwon, Young-Rip;Kim, Tae-Heung;Kim, Ji-Soo
    • Korean journal of applied entomology
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    • v.47 no.2
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    • pp.163-168
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    • 2008
  • Temperature dependent development of Tetranychus. urticae Koch was studied on the leaf of eggplant at 17, 22, 27, 32 and $37^{\circ}C$. T. urticae showed a minimum mortality at $27^{\circ}C$ and it increased at higher or lower temperatures than $27^{\circ}C$. The hatchability was low at 17 and $37^{\circ}C$. The duration of development decreased with increasing temperatures i.e., 5.3d at $37^{\circ}C$ and 25.8d at $17^{\circ}C$. Linear regression analysis of temperature vs. rate of development yielded the higher $r^2{\geq}0.88$ resulting in a good fit of the estimated line in the range of $17{\sim}37^{\circ}C$. Developmental zero temperature was $12.5^{\circ}C$ for the entire immature stage of female and $12.8^{\circ}C$ for that of male. Thermal constants were 80.5 and 74.7 degree days for those of female and male, respectively. Adult life span and oviposition period decreased with increasing temperatures. The number of eggs laid per female peaked at 141.0 eggs at $27^{\circ}C$, while that was a minimum 78.0 eggs at $37^{\circ}C$. Rate of hatchability, ratio of female, and $R_o$ were increased up to $27^{\circ}C$, and than declined thereafter. Intrinsic rate of natural increase (Rm) increased with rising temperatures and showed a maximum 0.5652 at $37^{\circ}C$. Also, ${\lambda}$ increased with increasing temperature. Doubling time (Dt) and generation time (T) decreased with increasing temperature.

A Meta Analysis of Using Structural Equation Model on the Korean MIS Research (국내 MIS 연구에서 구조방정식모형 활용에 관한 메타분석)

  • Kim, Jong-Ki;Jeon, Jin-Hwan
    • Asia pacific journal of information systems
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    • v.19 no.4
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    • pp.47-75
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    • 2009
  • Recently, researches on Management Information Systems (MIS) have laid out theoretical foundation and academic paradigms by introducing diverse theories, themes, and methodologies. Especially, academic paradigms of MIS encourage a user-friendly approach by developing the technologies from the users' perspectives, which reflects the existence of strong causal relationships between information systems and user's behavior. As in other areas in social science the use of structural equation modeling (SEM) has rapidly increased in recent years especially in the MIS area. The SEM technique is important because it provides powerful ways to address key IS research problems. It also has a unique ability to simultaneously examine a series of casual relationships while analyzing multiple independent and dependent variables all at the same time. In spite of providing many benefits to the MIS researchers, there are some potential pitfalls with the analytical technique. The research objective of this study is to provide some guidelines for an appropriate use of SEM based on the assessment of current practice of using SEM in the MIS research. This study focuses on several statistical issues related to the use of SEM in the MIS research. Selected articles are assessed in three parts through the meta analysis. The first part is related to the initial specification of theoretical model of interest. The second is about data screening prior to model estimation and testing. And the last part concerns estimation and testing of theoretical models based on empirical data. This study reviewed the use of SEM in 164 empirical research articles published in four major MIS journals in Korea (APJIS, ISR, JIS and JITAM) from 1991 to 2007. APJIS, ISR, JIS and JITAM accounted for 73, 17, 58, and 16 of the total number of applications, respectively. The number of published applications has been increased over time. LISREL was the most frequently used SEM software among MIS researchers (97 studies (59.15%)), followed by AMOS (45 studies (27.44%)). In the first part, regarding issues related to the initial specification of theoretical model of interest, all of the studies have used cross-sectional data. The studies that use cross-sectional data may be able to better explain their structural model as a set of relationships. Most of SEM studies, meanwhile, have employed. confirmatory-type analysis (146 articles (89%)). For the model specification issue about model formulation, 159 (96.9%) of the studies were the full structural equation model. For only 5 researches, SEM was used for the measurement model with a set of observed variables. The average sample size for all models was 365.41, with some models retaining a sample as small as 50 and as large as 500. The second part of the issue is related to data screening prior to model estimation and testing. Data screening is important for researchers particularly in defining how they deal with missing values. Overall, discussion of data screening was reported in 118 (71.95%) of the studies while there was no study discussing evidence of multivariate normality for the models. On the third part, issues related to the estimation and testing of theoretical models on empirical data, assessing model fit is one of most important issues because it provides adequate statistical power for research models. There were multiple fit indices used in the SEM applications. The test was reported in the most of studies (146 (89%)), whereas normed-test was reported less frequently (65 studies (39.64%)). It is important that normed- of 3 or lower is required for adequate model fit. The most popular model fit indices were GFI (109 (66.46%)), AGFI (84 (51.22%)), NFI (44 (47.56%)), RMR (42 (25.61%)), CFI (59 (35.98%)), RMSEA (62 (37.80)), and NNFI (48 (29.27%)). Regarding the test of construct validity, convergent validity has been examined in 109 studies (66.46%) and discriminant validity in 98 (59.76%). 81 studies (49.39%) have reported the average variance extracted (AVE). However, there was little discussion of direct (47 (28.66%)), indirect, and total effect in the SEM models. Based on these findings, we suggest general guidelines for the use of SEM and propose some recommendations on concerning issues of latent variables models, raw data, sample size, data screening, reporting parameter estimated, model fit statistics, multivariate normality, confirmatory factor analysis, reliabilities and the decomposition of effects.

Improvement of Radiosynthesis Yield of [11C]acetate ([11C]아세트산의 방사화학적 수율 증가를 위한 연구)

  • Park, Jun Young;Son, Jeongmin
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.2
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    • pp.74-78
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    • 2018
  • Purpose $[^{11}C]$acetate has been proved useful in detecting the myocardial oxygen metabolism and various malignancies including prostate cancer, hepatocellular carcinoma, renal cell carcinoma and brain tumors. The purpose of study was to improve the radiosynthesis yield of $[^{11}C]$acetate on a automated radiosynthesis module. Materials and Methods $[^{11}C]$acetate was prepared by carboxylation of grignard reagent, methylmagnesium chloride, with $[^{11}C]$$CO_2$ gas, followed by hydrolysis with 1 mM acetic acid and purification using solid phase extraction cartridges. The effect of the reaction temperature ($0^{\circ}C$, $10^{\circ}C$, $-55^{\circ}C$) and cyclotron beam time (10 min, 15 min, 20 min, 25 min) on the radiosynthesis yield were investigated in the $[^{11}C]$acetate labeling reaction. Results The maximum radiosynthesis yield was obtained at $-10^{\circ}C$ of reaction temperature. The radioactivities of $[^{11}C]$acetate acquired at $-10^{\circ}C$ reaction temperature was 2.4 times higher than those of $[^{11}C]$acetate acquired at $-55^{\circ}C$. Radiosynthesis yield of $[^{11}C]$acetate increased with increasing cyclotron beam time. Conclusion This study shows that radiosynthesis yield of $[^{11}C]$acetate highly dependent on reaction temperature. The best radiosynthesis yield was obtained in reaction of grignard reagent with $[^{11}C]$$CO_2$ at $-10^{\circ}C$. This radiolabeling conditions will be ideal for routine clinical application.

Influence of Hypoxia on Catecholamine Secretion Evoked by DMPP, McN-A-343, Excess $K^+$ and ACh from The Perfused Rat Adrenal Gland (저산소증이 흰쥐 관류부신에서 DMPP, McN-A-343, Excess $K^+$ 및 Ach의 카테콜아민 분비작용에 미치는 영향)

  • Lim Dong-Yoon;Heo Jae-Bong;Park Yoo Han
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.63-74
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    • 1995
  • It has been known that, during hypoxia, the adrenal medulla is activated to release catecholamines (CA) while hypoxia also inhibits high $K^+$ -induced CA secretion in the cultured bovine adrenal chromaffin cells. The present study was attempted to examine the effect of hypoxia on CA secretion evoked by chlinergic stimulation and membrane-depolarization from the isolated perfused rat adrenal glands and also to clarify its mechanism of action. For this purpose, using the isolated rat adrenal glands, the effects of hypoxia on CA release evoked by nicotinic ($N_1$) and muscarinic ($M_1$) receptor agonists, membrane-depolarizing agent, $Ca^{++}$-channel activator, intracellular $Ca^{++}$-releaser and ACh were determined. Experiments were carried out, perfusing Krebs solution pre-equilibrated with a gas mixture of 95% N_2$ and 5% $CO_2$. Hypoxia was maintained for $3{\sim}4$ hours through the experiments. Hypoxia gradually caused a time-dependent seduction in CA secretion evoked by DMPP ($100{\mu}M$), McN-A-343 ($100{\mu}M$), ACh (5.32 mM), Bay-K-8644 ($10{\mu}M$) and high $K^+$ (56 mM) respectively. How-ever, it did not affect CA secretion evoked by cyclopiazonic acid ($10{\mu}M$). Hypoxia itself also did fail to produce any influence on spontaneous secretory response of CA. These experimental results suggest that hypoxia depresses CA release evoked by both cholinergic stimulation and membrane-depolarization from the isolated rat adrenal medulla, and that this inhibitory activity may be due to the result of the direct inhibition of $Ca^{++}$ influx into the chromaffin cells without any effect on the calcium mobilization from the intracellular store.

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