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성충동약물치료 시행에 따른 소변 중 남성호르몬의 분석법 확립 (Validation of Analytical Method for Male Sex Hormone Monitoring in Urine due to the Chemical Castration)

  • 정수진;백승경;박선혜;손꽃잎;박용훈;이상기
    • 약학회지
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    • 제57권5호
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    • pp.330-336
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    • 2013
  • "The Act on Medication Treatment of Sexual Impulse of Sex Offenders" known as chemical castration has been effective since July 2011 in Korea. According to the law, monitoring of male sex hormone in urine is enforced to request National Forensic Service more than once a month after injection of medicine designed to reduce sex impulse. We established a rapid and sensitive method for the monitoring of testosterone (T) and epitestosterone (E) in human urine by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Three mL of urine was pretreated by solid-phase extraction for purification and performed enzymatic hydrolysis. The pretreated samples were extracted twice with 2 ml of ethyl acetate and n-hexane (2 : 3). The separation was applied on Thermo Hypersil GOLD C18 column ($1.9{\mu}m$, $100{\times}2.1mm$). A gradient elution of methanol and water of 0.1% formic acid were used as mobile phase and the retention time was less than 10 min. LC-MS/MS system coupled with an electrospray ionization source was performed in multiple reaction monitoring mode. The transitions of the analytes executed as following: m/z $289{\rightarrow}97$, 109 for T and E, m/z $292{\rightarrow}109$ for $T-d_3$ and $E-d_3$ as internal standards. The validation results of the method were satisfactory. The limits of detection were 0.05 ng/ml and the limits of quantification were 0.1 ng/ml. This method was successfully applied to real human urine sample. The developed method will be useful for monitoring T/E ratio in urine of sex offenders.

Changes in the Levels of Insulin-like Growth Factors (IGF-I and IGF-II) in Bovine Milk According to the Lactation Period and Parity

  • Kang, S.H.;Kim, J.U.;Kim, Y.;Han, K.S.;Lee, W.J.;Imm, J.Y.;Oh, S.;Park, D.J.;Moon, Y.I.;Kim, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권1호
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    • pp.119-123
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    • 2007
  • The objectives of this study were to characterize the changes occurring in the levels of insulin-like growth factors (IGF-I and IGF-II) in bovine milk during a one-year lactation period, and to determine the parameters affecting IGF content in bovine milk. Milk was collected individually from lactating Holstein cows (n=70), and IGF-I and -II levels were determined via radioimmunoassay, using 125I after acid-ethanol treatment. The proximate compositions of the milk samples were determined using a near-infrared milk analyzer. The data were analyzed by the GLM and CORR procedures using SAS software to determine significant differences (p<0.05) occurring within groups (dairy farms, lactation periods, season, and parity). We noted an approximately six-fold reduction in the IGF-I concentration (from 2,462.7 to 353.0 ng/ml) and a three-fold drop in the IGF-II concentration (from 929.1 to 365.7 ng/ml) in the bovine colostrum, between 6 h after parturition and 18 h after parturition. IGF-I and -II content, measured at the early, middle, and late stages of lactation did not change significantly throughout the entirety of the lactation period. Interestingly, parity did not significantly affect IGF-I content, but did significantly affect IGF-II content between the primiparous and multiparous cows. We also found there were no significant relationships between IGF-I and total protein content or somatic cell counts (p<0.05).

Extraction of Phthalate Esters in Environmental Water Samples Using Layered-Carbon Magnetic Hybrid Material as Adsorbent Followed by Their Determination with HPLC

  • Wang, Weina;Wu, Qiuhua;Zang, Xiaohuan;Wang, Chun;Wang, Zhi
    • Bulletin of the Korean Chemical Society
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    • 제33권10호
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    • pp.3311-3316
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    • 2012
  • In this paper, a layered-carbon-$Fe_3O_4$ (LC-$Fe_3O_4$) hybrid material was synthesized through a facile one-pot solvothermal method and used as the adsorbent for the preconcentration of some phthalate esters (dimethyl phthalate, diethyl phthalate, diallyl phthalate, diisobutyl phthalate and benzyl butyl phthalate) in water samples. The effects of the adsorbent dosage, extraction time, the solution pH and salinity on the adsorption of the phthalate esters (PAEs) were investigated. The magnetic nanocomposite adsorbent could remove and enrich the PAEs from water samples efficiently. After the adsorption, the analytes were desorbed and then determined by high performance liquid chromatography-ultraviolet detection. Under the optimum conditions, the enrichment factors of the method for the analytes were in the range from 161 to 180. A linear response with peak area as the quantification signal was observed in the concentration range from 0.5 to $100ng\;mL^{-1}$. The limits of detection (S/N = 3) of the method were between 0.08 and $0.1ng\;mL^{-1}$. The method was suitable for the determination of trace phthalate esters in environmental water samples.

배양된 사람치은각화상피세포의 미세구조 (CULTURED HUMAN ORAL KERATINOCYTES; ULTRASTRUCTURAL STUDY)

  • 권용대;이백수;주성숙
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제21권3호
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    • pp.231-239
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    • 1999
  • In oral and maxillofacial surgery, there are many cases requiring the graft of epidermal tissues such as maxillectomy, and vestibuloplasty. There have been so many challenges for the culture of the epidermal tissue. Observing the ultrastructure of the cultured human oral kertinocytes, we could compare this findings with that of in vivo ones. With that, we could find the differencies and similarities between cultured cells and in vivo ones, and evaluate the clinical applications of cultured tissue. Human gingiva was obtained and the specimen was explanted on 24-well plate. Two types of culture media were used in this culture system. One was for the growth of the keratinocytes (Media I), and the other was for the stratification (Media II). Media I had special ingredients for the epidermal growth. Those were 0.5% dimethyl sulfoxide (DMSO), 30ng/ml of epidermal growth factor (EGF), 30ng/ml of cholera toxin, and $5{\mu}g/ml$ of transferrin. We cultured the oral keratinocytes for 3 weeks, and at that time the cultured keratinocytes were processed to prepare the specimen for the TEM study. The results were as follows.; 1. In the phase contrast micrograph, epidermal outgrowth firstly appeared on the 3rd day after explantation, and the growing keratinocytes were activley mitotic, and had polygonal shape and increased N/C ratio. 2. In the phase contrast micrograph, the outer most cells exhibited areas where broad cytoplasmic processes extended out onto the culture subtratum(fan-like appaearance). 3. In the TEM micrographs, the cultured keratinocytes showed stratification. The cells were in elongated form, and there were no morphologic differencies among the layers usually found in the in vivo gingiva. 4. Most of cellular organelles underwent lysis, and keratohyaline granules were seen. Tonofibrils were dispersed in the cytoplasm. 5. The cells were interconnected by desmosomes, and their frequency of distribution was considered to be lower than that of in vivo keratinocytes. 6. We could conclude the cultured oral keratinocytes exhibited signs of terminal differentiation.

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청소년의 신체활동 강도가 비타민 D 결핍에 미치는 영향 (The Effect of the Strength of Physical Activity on Vitamin D Deficiency among Korean Adolescents)

  • 김지윤;최숙자;이윤정
    • 한국학교보건학회지
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    • 제30권2호
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    • pp.146-153
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    • 2017
  • Purpose: The purpose of this study was to identify the association between physical activity and vitamin D deficiency status among Korean adolescents. Methods: This is cross-sectional analysis of Korean adolescents aged 10-18 years from the 5th Korea National Health and Nutrition Examination Survey(KNHANES 2010-2012; n=2,384). We estimated the prevalence of vitamin D deficiency and identified the correlations with the strength of physical activity. $x^2$ test, t-test, and multiple logistic regression using complex sample analysis were done. The odds ratio of vitamin D deficiency by physical activity was calculated using complex sample multivariate logistic regression analysis. Results: The prevalence of vitamin D deficiency was 78.2%. The mean serum 25 (OH) D level in deficiency group and non-deficiency group was 14.4ng/mL, 23.7ng/mL respectively. Differences by age strata (the prevalence ranging from 68.5% to 86.3%) and gender (78.6% for boy and 84.5% for girl) were identified. Among adolescents aged 16-18 or girl in vigorous physical activity, a significant correlation between vitamin D deficiency and physical activity was observed, whereas no significant correlation were found among adolescents in moderate physical activity. Conclusion: In conclusion, vigorous physical activity was associated with vitamin D deficit in Korean adolescents. Thus, intervention programs enhancing vigorous activity than moderate activity need to be developed in schools and community.

LC-MS/MS를 이용한 담배 연기 중 Heterocyclic Amines의 분석 (Analysis of Heterocyclic Amines in Mainstream Cigarette Smoke using by LC-MS/MS)

  • 김익중;장기철;지상운;민혜정;김효근;황건중
    • 한국연초학회지
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    • 제30권1호
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    • pp.33-38
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    • 2008
  • This study was carried out to determine the analytical methods for heterocyclic amines(HAs) of the tobacco smoke by LC/MS/MS. HAs have been found in pyrolysate of protein and cooked food including protein, were known the Sugimura compound. HAs content of the smoke were known to exist very low ppb level. Especially, some of HAs are mutagenic and carcinogenic compounds. In according to IARC, the toxicity of N-heterocyclic amines classified IARC class 2A or 2B group. Precursors of these compounds are glutamic acid, protein and free amino acids including tryptophan, therefore, the precursors have been proved in cooked food continuously. This study was investigate multiple analysis methods for HAs and HAs contents of some commercial products. In this study, we used the linear type smoking machine for HAs analysis. At the ISO conditions, mainstream smoke was collected on cambridge filter pad, and then cambridge filter pad was extracted by 0.1% acetic acid. The extracted solution were passed cation exchange SPE cartridge to remove matrix, samples were analyzed using LC/MS/MS on MRM mode. From the result that optimized this methods, the correlation coefficient(R) of the individual compounds were good linearity over 0.999, recovery rate over 96% and the limit of detection were good values between 0.06 to 0.37 ng/mL, In addition, HAs content of some commercial products were in range of 0.02 to 43.8 ng/cig.

Mometasone Furoate Suppresses PMA-Induced MUC-5AC and MUC-2 Production in Human Airway Epithelial Cells

  • Poachanukoon, Orapan;Koontongkaew, Sittichai;Monthanapisut, Paopanga;Pattanacharoenchai, Napaporn
    • Tuberculosis and Respiratory Diseases
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    • 제80권1호
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    • pp.60-68
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    • 2017
  • Background: Mucus hypersecretion from airway epithelium is a characteristic feature of airway inflammatory diseases. Tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) regulates mucin synthesis. Glucocorticoids including mometasone fuorate (MF) have been used to attenuate airway inflammation. However, effects of MF on mucin production have not been reported. Methods: Effects of MF and budesonide (BUD) on the phorbol-12-myristate-13-acetate (PMA)-induction of mucin and TNF-${\alpha}$ in human airway epithelial cells (NCI-H292) were investigated in the present study. Confluent NCI-H292 cells were pretreated with PMA (200 nM) for 2 hours. Subsequently, the cells were stimulated with MF (1-500 ng/mL) or BUD (21.5 ng/mL) for 8 hours. Dexamethasone ($1{\mu}g/mL$) was used as the positive control. Real-time polymerase chain reaction was used to determine MUC2 and MUC5AC mRNA levels. The level of total mucin, MUC2, MUC5AC, and TNF-${\alpha}$ in culture supernatants were measured using enzyme-linked immunosorbent assay. Results: MF and BUD significantly suppressed MUC2 and MUC5AC gene expression in PMA-stimulated NCI-H292 cells. The inhibitory effects of the two steroid drugs were also observed in the production of total mucin, MUC2 and MUC5AC proteins, and TNF-${\alpha}$. Conclusion: Our findings demonstrated that MF and BUD attenuated mucin and TNF-${\alpha}$ production in PMA-induced human airway epithelial cells.

저온열탈착기술을 이용한 에틸렌 및 저분자 탄화수소 분석방법 연구 (A Study of Analytical Method for Ethylene and Low Weight Hydrocarbons (LWHC) using Thermal Desorption and Gas Chromatography-Flame Ionization Detector with (TD-GC-FID))

  • 김보원;김기현;김용현;안정현
    • 한국대기환경학회지
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    • 제30권1호
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    • pp.77-87
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    • 2014
  • In this study, an experimental approach to measure a suite of low weight hydrocarbons was investigated with an emphasis on ethylene (EL) along with many others (ethane (EA), propane (PA), propylene (PL), n-butane (BA), acetylene (AL), methyl acetylene (ML)). Their concentrations were quantified using GC-FID system equipped with thermal desorption (TD) system. The TD-based analysis was conducted using both Link Tube/Thermal Desorber (LT/TD) method and Modified Injection through a Thermal Desorption (MITD) method. The results of these analyses were evaluated in a number of respects. The system allowed the detection of all compounds except methane with the mean response factor (RF) of 10.28 (EA) to 11.94 (PL). The method detection limits of target compounds were seen in the range of 0.027 (ML) to 0.146 ng (BA). The emission flux of some environmental samples (fruits), when measured using a small flux chamber system, fell in the range of 0.14 (AL: Kiwi) to $181ng{\cdot}g^{-1}{\cdot}hr^{-1}$ (EL: Apple Peel). The results of this study confirm that the experimental approach developed in this study allows to accurately measure emissions of low weight hydrocarbons (LWHC) like ethylene from various natural and man-made source processes.

유도결합플라즈마 질량분석법에 의한 납 취급 근로자와 일반인의 혈장 중 납 분석 및 상관성 분석 (Analysis of Trace Level and Correlation of Lead in the Plasma of Field Workers and General Public by ICP-MS)

  • 이성배;양정선;최성봉;김남수;이병국;신호상
    • 한국산업보건학회지
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    • 제23권2호
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    • pp.65-74
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    • 2013
  • Objectives: This study attempted to develop a method to measure ultra-trace lead concentrations in plasma using Inductively Coupled Plasma Mass Spectrometry(ICP-MS) and to test whether plasma lead can be used as a biomarker for the biological monitoring of exposure to lead. Methods: Lead concentrations in 160 plasma samples of field workers and 42 plasma samples from the control group were measured by ICP-MS. Blood zinc protophorphyrin(ZPP) concentrations and urinary ${\delta}$-aminolevulinic acid${\delta}-ALA$) were measured for correlation analysis with plasma lead. Results: The mean lead level in the plasma of the workers exposed to lead at work were 786.1 ng/L. Plasma lead levels were not correlated with blood ZPP or urinary ${\delta}-ALA$ concentrations. Otherwise, plasma lead levels showed a good correlation coefficient of 0.400 with blood lead levels, and their correlation coefficient had a better value of 0.552 for the non-smoking and drinking group. In the general population group which was not exposed to lead in the workplace and was considered the control group, the mean concentration of plasma lead was 123.1 ng/L. The plasma lead levels for the general population group showed a good correlation coefficient of 0.520 with blood ZPP and urinary ${\delta}-ALA$ concentrations.

건폐탕(健肺陽)이 호흡기 뮤신의 생성 및 유전자 발현에 미치는 영향 (Effect of Geonpye-tang(GPT) on Production and Gene Expression of Respiratory Mucin)

  • 정병진;김호;서운교
    • 대한한방내과학회지
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    • 제30권4호
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    • pp.685-695
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    • 2009
  • Objectives : In this study, the author tried to investigate whether Geonpye-tang(GPT) significantly affects PMA-, EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells. Materials and Methods : Effects of the agent on PMA-, EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of GPT and treated with PMA (10ng/ml) or EGF (25ng/ml) or TNF-alpha (0.2nM), to assess both effect of the agent on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicity of the agent was assessed by examining the rate of survival and proliferation of NCI-H292 cells after treatment with the agent over 72 hrs (SRB assay). Results : (1) GPT significantly inhibited PMA-induced and EGF-induced MUC5AC mucin production from NCI-H292 cells. However, GPT did not affect TNF-alpha-induced MUC5AC mucin production. (2) GPT significantly inhibited the expression levels of PMA-, EGF- or TNF-alpha-induced MUC5AC genes in NCI-H292 cells (3) GPT did not show significant cytotoxicity to NCI-H292 cells. Conclusion : This result suggests that GPT can affect the production and gene expression of respiratory mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion. This can explain the traditional use of GPT in oriental medicine. Effects of GPT with their components should be further investigated using animal experimental models that reflect pathophysiology of airway diseases through future studies.

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