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Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
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    • v.31 no.4
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    • pp.161-170
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    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

The Effects of Air-borne Particulate Matters on the Alveolar Macrophages for the TNF-α and IL-1β Secretion (미세분진이 흰쥐의 폐포대식세포에서 TNF-α와 IL-1β의 형성에 미치는 효과)

  • Li, Tian Zhu;Lee, Soo-Jin;Park, Se-Jong;Chang, Byung-Joon;Lee, Jong-Hwan;Kim, Kil-Soo;Lee, Myoung-Heon;Choe, Nong-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.5
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    • pp.554-563
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    • 2006
  • Background: PM is known to induce various pulmonary diseases, including asthma, cancer, fibrosis and chronic bronchitis. Despite the epidemiological evidence the pathogenesis of PM-related pulmonary diseases is unclear. Methods: This study examined the effects of PM exposure on the secretion of $TNF-{\alpha}$ and $IL-1{\beta}$ in the cultured alveolar macrophages. The cultured primary alveolar macrophages were treated with the medium, PM ($5{\sim}20{\mu}g/cm^2$), LPS (5ng/ml), and PM with LPS for 24h and 48h respectively. ELISA was used to assay the secreted $TNF-{\alpha}$ and $IL-{\beta}$ in the culture medium. Western blotting was used to identify and determine the level of proteins isolated from the culture cells. The cells cultured in the $Lab-Tek^{(R)}$ chamber slides were stained with immunocytochemical stains. Results: PM induced $TNF-{\alpha}$ and $IL-1{\beta}$ secretion in the culturing alveolar macrophages, collected from the SPF and inflammatory rats. However, the effects were only dose-dependent in the inflammatory macrophages. When the cells were co-treated with PM and LPS, there was a significant synergistic effect compared with the LPS in the both cell types. Conclusion: PM might be play an important role in the induction and/or potentiation of various lung diseases by oversecretion of $TNF-{\alpha}$ and $IL-1{\beta}$.

Reproductive Ecology of the Silver Pomfret Pampus argenteus on the West Coast of Korea (한국 서해산 병어, Pampus argenteus의 번식생태)

  • Chung, Ee-Yung;Bae, Joo-Seung;Kang, Hee-Woong;Lee, Hwang-Bok;Lee, Ki-Young
    • Development and Reproduction
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    • v.12 no.2
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    • pp.169-181
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    • 2008
  • Reproductive ecology of the silver pomfret, Pampus argenteus were investigated by histological observations and morphometric data. Samples were collected by the stow net at the coastal area of Jaun-Do, Muan-gun, Korea, from January to December, 2006. P. argenteus is dioecious, the ovary is composed of many ovarian lobules, showing a pair of saccular structure, and the testis is composed of many seminiferous lobules, showing a pair of lobular structure. From February (growing stage) to September (after spawning), monthly changes in the gonadosomatic index, hepatosomatic index, and condition factor in females and males showed similar patterns with the gonad developmental phases. Judging from the results of their indice, it is assumed that spawning in females and males occur from May to July. The reproductive cycle can be classified into five successive stages in females: early growing stage (February to March), late growing stage (March to April), mature stage (March to July), ripe and spent stage (May to July), and recovery and resting stage (July to February); in males, the cycle can be divided into four successive stages: growing stage (February to April), mature stage (March to June), ripe and spent stage (May to July), and recovery and resting stage (July to February). According to the frequency distributions of egg diameters in the breeding season, P. argenteus is presumed to be spring-summer spawning species and polycyclic species to spawn 2 times or more during one spawning season. Number of total eggs in absolute fecundity were proportional to body length and body weight, respectively. Number of total eggs in absolute fecundity per body weight were also proportional to the body length, but if the increase of body weight considerably increased, rather total eggs in relative fecundity decreased with the increase of body weight. Percentage of first sexual maturity of P. argenteus were over 50% in females and males of 12.1 to 15.0 cm in body length, and 100% for fishes over 18.1 cm in length. Therefore, both sexes were regarded to be sexually mature at one year of age.

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Protective Effect of the Inhibition of PAF Remodeling and Adhesion Molecule on the Oxidative Stress of the Lungs of Rats Given Endotoxin Intratracheally (내독소에 의해 유도된 급성 폐 손상에서 PAF Remodelling 및 Adhesion Molecule의 억제가 폐장내 Oxidative Stress에 미치는 영향)

  • Shin, Tae Rim;Na, Bo Kyung;Lee, Young Man
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.3
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    • pp.276-284
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    • 2005
  • Backgraound : There have been many reports on the pathogenesis of sepsis-induced acute respiratory distress syndrome(ARDS) but, the precise mechanism has not been elucidated. This study examined the protective effect of an inhibition of platelet activating factor(PAF) remodeling and the adhesion molecule on the oxidative stress of the lungs in rats with an endotoxin induced acute lung injury(ALI). Methods : ALI was induced in Sprague-Dawley rats by instilling an E-coli endotoxin into the trachea. Ketotifen and fucoidan were used respectively to inhibit PAF remodeling and adhesion molecule. The lung leak index, lung myeloperoxidase(MPO) activity, bronchoalveolar lavage(BAL) fluid neutrophil count and lyso PAF acetyltransferase activity(AT), were measured and an ultrastructural study and cytochemical electron microscopy were performed. Results : The lung leak index, lung MPO activity, BAL fluid neutrophil count and lyso PAF AT activity was higher in the endotoxin-treated rats. In addition, severe destruction of the pulmonary architecture and increased hydrogen peroxide production were identified. These changes were reversed by ketotifen. However, fucoidan did not appear to have any protective effects. Conclusion : The inhibition of PAF remodeling appeared to be effective in decreasing the endotoxin-induced ALI. In addition, this effect might be derived from the inhibition of neutrophilic oxidative stress. However, the inhibition of the adhesion molecules by fucoidan appeared to be ineffective in decreasing the endotoxin-induced ALI.

Long-tenn Evaluation of Extruded Pellet Diets Compared to Raw Fish Moist Pellet Diet for Growing Flounder, Paralichthys olivaceus (넙치 미성어 건조 배합사료 및 습사료의 장기사육 평가)

  • Kim Kang-Woong;Kang Yong Jin;Kim Kyong-Min;Lee Hae Young;Kim Kyoung-Duck;Bai Sungchul C.
    • Journal of Aquaculture
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    • v.18 no.4
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    • pp.225-230
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    • 2005
  • This experiment was conducted to compare the effects of extruded pellets and raw fish-based pellet on olive flounder Paralichthys. olivaceus. Six diets were prepared for this study: two formulated extruded pellets (FEP1 & FEP2), three commercially available extruded pellets (CEP1, CEP2 & CEP3) and moist pellet (MP). Weight gain offish fed FEP1 and CEP3 were significantly higher (P<0.05) than those of fish fed FEP2, CEP1, CEP2 and MP, while that of fish fed MP was not significantly different (f<0.05) from those of fish fed the FEP2, CEP1 and CEP2. Feed efficiency of fish fed CEP2 was significantly lower (P<0.05) than those of fish fed FEP1, FEP2, CEP1, CEP3 and MP. There was no significant difference in protein efficiency ratio and hepatosomatic index between fish fed FEP1 and CEP3, and among fish fed FEP2, CEP1 and CEP2. There was no significant difference in condition factor among fish fed the FEP1 and CEP3, and between fish fed FEP2, CTP1 and MP. However, fish fed MP had a lower survival rate than fish fed the other five EP These results suggest that diet FEPl could be developed to replace MP for the owing stage of flounder without adverse effects on growth performance.

Anti-Obesity Effect of Pine Cone (Pinus koraiensis) Supercritical Extract in High-Fat Diet-Induced Obese Mice (고지방식이로 유도한 비만 Mice에서 잣송이 초임계 추출물의 항비만 효과)

  • Lee, Dasom;Lee, Minhee;Kim, Hyesook;Jeong, Tuk-Rai;Yang, Hyun-Pil;Hyun, Heo Seok;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.12
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    • pp.1701-1707
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    • 2016
  • The present study investigated the anti-obesity effect of pine cone (PC, Pinus koraiensis) supercritical extract in high-fat diet (HFD)-induced obese mice. Male C57BL/6J mice were treated with HFD, HFD+catechin, and HFD+PC [two different doses, 20 mg/kg body weight (b.w.) and 100 mg/kg b.w.] in each AIN93G supplement for 8 weeks. Treatment of HFD mice with both low and high doses of PC significantly reduced body weight gain compared to HFD mice. Liver weight of mice was reduced in both the low and high dose PC-supplemented groups (24.19% and 19.83%, respectively). Total adipose tissue weight of mice was reduced in both the low and high dose PC-supplemented groups (45.54% and 62.66%, respectively). Serum total cholesterol, triglyceride, LDL cholesterol, and HDL cholesterol were reduced in the low and high dose PC-supplemented groups, and ratios of HDL cholesterol to LDL cholesterol increased by 94.55% in the high dose PC-supplemented group. Serum leptin was significantly reduced in the low and high dose PC-supplemented groups (28.14% and 62.72%, respectively). These results were supported by genetic expression of protein and enzymes related to lipid metabolism assessed by real-time PCR. There was significant reduction of lipid regulatory transcription factors such as $PPAR-{\gamma}$, C/EBP, and SREBP and lipid enzymes such as fatty acid synthesis and lipoprotein lipase in the low and high dose PC-supplemented groups. However, there was no statistical difference between low and high dose PC treatments. These results suggest that pine cone supercritical extract supplementation is able to regulate serum lipid profiles by reducing total cholesterol, triglyceride, and LDL cholesterol levels, followed by regulation of expression of lipid metabolic factors, resulting in reduction of weight gain in HFD-induced obese mice.

Characteristics and Risk Assessment of Heavy Metals in the Stormwater Runoffs from Industrial Region Discharged into Shihwa Lake (시화호 산업지역 강우유출수 내 중금속 유출특성 및 위해성 평가)

  • Ra, Kongtae;Kim, Joung-Keun;Lee, Jung-Moo;Lee, Seung-Yong;Kim, Eun-Soo;Kim, Kyung-Tae
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.17 no.4
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    • pp.283-296
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    • 2014
  • The distribution of heavy metals in the stormwater runoffs from industrial sites around Shihwa Lake that implements the total pollutant load management system (TPLMS) was studied to characterize the temporal changes of metal concentrations and to assess the ecological risk in dissolved and particulate phases of the selected metals. The dissolved Co and Ni concentration demonstrated first flush and tended to decrease with increasing of the duration of rainfall. The intensity of precipitation was found to be the main controlling factor of particulate metals in the stromwater runoffs. The particulate concentration of Pb accounted for 97.2% so the particulate phase was its main form. Other metals followed the sequence: Pb>Cu>Cd>Co>Zn>Ni. The particulate-dissolved partitioning coefficient ($K_d$) indicated that the $K_d$ of Pb were bigger than that of other metals because the metal Pb in the stormwater runoffs is quickly removed into the particulate phase. In a single day rainfall event, total runoff fluxes for total metals as the sum of dissolved and particulate forms through only two sewer outlets were 2.21 kg for Co, 30.5 kg for Ni, 278.3 kg for Cu, 398.3 kg for Zn, 0.39 kg for Cd and 40.0 kg for Pb, respectively. Given the annual rainfall, the number of rain days and the basin area for total pollutant load management system (TPLMS) of Shihwa area, enormous amount of non-point metal pollutants were entered into Lake with any treatment. The dissolved metals (e.g., Ni, Cu and Zn) in the stormwater runoffs exceeded the acute water quality criteria. Additionally, all metals were significantly enriched in the particulate phase and exceeded the PEL criteria of sediment quality guidelines (SQGs). These results indicated that the heavy metals in the stormwater runoffs may pose a very high ecological risk to the coastal environments and ecosystem.

Soil Fertility Evaluation with Adoption of Soil Map Database for Tobacco Fields (토양도 자료를 활용한 연초 경작지의 비옥도 평가)

  • Hong, Soon-Dal;Park, Hyo-Taek
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.2
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    • pp.95-108
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    • 1999
  • Field experiments were conducted in the 101 tobacco fields(51 fields in 1985 and 50 fields in 1986) of chief tobacco producing counties of Chungbuk province(Jincheon, Eumseong, Goesan, and Joongweon counties), Chungnam province(Cheonweon county), and Kyongbuk province (Cheongdo, Seongju, and Andong counties) for two years from 1985 to 1986 in order to evaluate soil fertility using chemical properties and soil map database. Pot experiments also on the same soils were conducted and the results were compared to those of field experiments. The yield of tobacco in the plots of no fertilization was considered as a basic factor representing the soil fertility and was evaluated by nineteen independent variables, that was 9 chemical properties and 10 soil map databases. These independent variables were classified into two groups, 11 quantitative indexes and 9 qualitative indexes, and were analyzed by multiple linear regression(MLR) of SAS by REG and GLM models. The yield of tobacco in the plot of no fertilization showed high variations, e.g. the difference between minimum and maximum yields was about 5.0-5.5 times in the pot experiment and 8.2-14.9 times in the field experiment. The indexes indicating close link between yield of tobacco and soil chemical indexes, was selected but it was not well matched by the years or between pot and field experiments. Also, the standardized partial regression coefficients of quantitative indexes for the yield of field were less than 1.0, suggesting that it is difficult to develop an available single index for the evaluation of soil fertility. Evaluation for the soil fertility of field by MLR was better than that of single regression and it was gradually improved by adding chemical properties, quantitative indexes, and qualitative indexes of soil map. For example, the coefficient of determination ($R^2$) of MLR for the yield of 1985 was increased to 0.422 with chemical indexes, 0.503 by addition of quantitative indexes, and 0.633 by the additional adding of qualitative indexes of soil map, compared to 0.244 of single index, $NO_3-N$ content of soil. Consequently, it is assumed that this approach by MLR with quantitative and qualitative indexes including chemical properties and soil map databases was available as an evaluation model of soil fertility for tobacco field.

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Proving Causation With Epidemiological Evidence in Tobacco Lawsuits (담배소송에서 역학적 증거에 의한 인과관계의 증명에 관한 소고)

  • Lee, Sun Goo
    • Journal of Preventive Medicine and Public Health
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    • v.49 no.2
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    • pp.80-96
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    • 2016
  • Recently, a series of lawsuits were filed in Korea claiming tort liability against tobacco companies. The Supreme Court has already issued decisions in some cases, while others are still pending. The primary issue in these cases is whether the epidemiological evidence submitted by the plaintiffs clearly proves the causal relationship between smoking and disease as required by civil law. Proving causation is difficult in tobacco lawsuits because factors other than smoking are involved in the development of a disease, and also because of the lapse of time between smoking and the manifestation of the disease. The Supreme Court (Supreme Court Decision, 2011Da22092, April 10, 2014) has imposed some limitations on using epidemiological evidence to prove causation in tobacco lawsuits filed by smokers and their family members, but these limitations should be reconsidered. First, the Court stated that a disease can be categorized as specific or non-specific, and for each disease type, causation can be proven by different types of evidence. However, the concept of specific diseases is not compatible with multifactor theory, which is generally accepted in the field of public health. Second, when the epidemiological association between the disease and the risk factor is proven to be significant, imposing additional burdens of proof on the plaintiff may considerably limit the plaintiff's right to recovery, but the Court required the plaintiffs to provide additional information such as health condition and lifestyle. Third, the Supreme Court is not giving greater weight to the evidential value of epidemiological study results because the Court focuses on the fact that these studies were group-level, not individual-level. However, group-level studies could still offer valuable information about individual members of the group, e.g., probability of causation.

Glycoprotein Isolated from Morus indica Linne Has an Antioxidative Activity and Inhibits Signal Factors Induced by Bisphenol A in Raw 264.7 Cells (뽕잎 당단백질의 항산화능과 Raw 264.7 세포에 있어서 bisphenol A에 유도된 신호전달인자의 억제)

  • Shim, Jae-Uoong;Lee, Sei-Jung;Oh, Phil-Sun;Lim, Kye-Taek
    • Korean Journal of Food Science and Technology
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    • v.39 no.2
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    • pp.209-216
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    • 2007
  • The present study investigated anti-oxidative and anti-inflammatory activity of glycoprotein isolated from Morus Indica Linne (MIL glycoprotein). We found that MIL glycoprotein has a molecular weight of 32 kD and consists of carbohydrate (40.03%) and protein (59.97%), and that it has a strong scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical $({\cdot}OH)$, and superoxide anion $(O_2{\cdot}\;^-)$ radicals. In addition, MIL glycoprotein had a stable character and an optimal DPPH radical scavenging activity in the alkaline and neutral pH solution, and up to at 105. However, the results indicated that it has a minimal scavenging activity in the metal ionic solution ($Ca^{2+}$, $Mn^{2+}$, and $Mg^{2+}$) in the presence of EDTA. In addition, we further investigated whether MIL glycoprotein scavenges oxygen radicals and blocks inflammation-related signals in the bisphenol A (BPA)-stimulated Raw 264.7 cells. The results in this study showed that it has a character to scavenge the productions of reactive oxygen species (ROS) and nitric oxide (NO) dose-dependently. Also it blocked the activities of inflammation-related signals such as nuclear factor-kappa B ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS). For example, it had an inhibitory effect on the activation of $NF-{\kappa}B$ (p50) and iNOS proteins at 200 ${\mu}g/mL$ MIL glycoprotein. Here, we speculate that MIL glycoprotein is one of natural antioxidants and of modulators of the BPA-induced inflammation.