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Effect of Verapamil on Cellular Uptake of Tc-99m MIBI and Tetrofosmin on Several Cancer Cells (수종의 암세포에서 Verapamil이 Tc-99m MIBI와 Tetrofosmin의 섭취에 미치는 영향)

  • Kim, Dae-Hyun;Yoo, Jung-Ah;Suh, Myung-Rang;Bae, Jin-Ho;Jeong, Shin-Young;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.85-98
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    • 2004
  • Purpose: Cellular uptake of $^{99}mTc$-sestamibi (MIBI) and $^{99}mTc$-tetrofosmin (TF) is low in cancer cells expressing multidrug resistance(MDR) by p-glycoprotein(Pgp) or multidrug related protein(MRP). Verapamil is known to increase cellular uptake of MIBI in MDR cancer cells, but is recently reported to have different effects on tracer uptake in certain cancer cells. This study was prepared to evaluate effects of verapamil on cellular uptake of MIBI and TF in several cancer cells. Materials and Methods: Celluar uptakes of Tc-99m MIBI and TF were measured in erythroleukermia K562 cell, breast cancer MCF7 cell, and human ovarian cancer SK-OV-3 cells, and data were compared with those of doxorubicin-resistant K562(Ad) cells. RT-PCR and Western blot analysis were used for the detection of mdr1 mRNA and Pgp expression, and to observe changes in isotypes of PKC enzyme. Effects of verapamil on MIBI and TF uptake were evaluated at different concentrations upto $200{\mu}M\;at\;1{\times}10^6\;cells/ml\;at\;37^{\circ}C$. Radioactivity in supernatant and pellet was measured with gamma counter to calculate cellular uptake ratio. Toxicity of verapamil was measured with MTT assay. Results: Cellular uptakes of MIBI and TF were increased by time in four cancer cells studied. Co-incubation with verapamil resulted in an increase in uptake of MIBI and TF in K562(Adr) cell at a concentration of $100{\mu}M$ and the maximal increase at $50{\mu}M$ was 10-times to baseline. In contrast, uptakes of MIBI and TF in K562, MCF7, SK-OV3 cells were decreased with verapamil treatment at a concentration over $1{\mu}M$. With a concentration of $200{\mu}M$ verapamil, MIBI and TF uptakes un K562 cells were decreased to 1.5 % and 2.7% of those without verapamil, respectively. Cellular uptakes of MIBI and TF in MCF7 and SK-OV-3 cells were not changed with $10{\mu}M$, but were also decreased with verapamil higher than $10{\mu}M$, resulting 40% and 5% of baseline at $50{\mu}M$. MTT assay of four cells revealed that K562, MCF7, SK-OV3 were not damaged with verapamil at $200{\mu}M$. Conclusion: Although verapamil increases uptake of MIBI and TF in MDR cancer cells, cellular uptakes were further decreased with verapamil in certain cancer cells, which is not related to cytotoxicity of drug. These results suggest that cellular uptakes of both tracers might differ among different cells, and interpretation of changes in tracer uptake with verapamil in vitro should be different when different cell lines are used.

Resolution of the Triacylglycerols Containing Conjugate Trienoic Acids into Their Molecular Species by HPLC in the Reversed-phase and Silver Ion Mode (Reversed-phase 및 $Ag^{+}$-HPLC에 의한 Conjugate Trienoic Acid 함유(含有) Triacylglycerol 분자종(分子種)의 상호분리(相互分離))

  • Kim, Seong-Jin;Woo, Hyo-Kyeng;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.18 no.3
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    • pp.197-213
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    • 2001
  • Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

Studies on the Effects of Caponization and Various Hormone Treatment on the Meat Production and Quality in Growing Chicken (닭에 있어서 거세(去勢) 및 Hormone 처리(處理)가 산육성(産肉性) 및 육질(肉質)에 미치는 영향(影響)에 관한 연구(硏究))

  • Ra, Kwang Yon
    • Korean Journal of Agricultural Science
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    • v.2 no.1
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    • pp.9-47
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    • 1975
  • These experiments were caried out to study the effects of caponization and various hormone treatments upon meat production and improvement of meat quality of growing chicken. Sixtyseven days old 160 New Hampshire cockerels were treated and growth rate, carcass yield, change of weight of individual organs, meat composition and change of amino acid were measured and analysed. Otherwise change of testis and thyroid gland by hormone treatment were investigated histologically. The results obtained were as follows. 1. The effectst of caponization and hormone treatment upon meat production were; 1) Body weight of cockerels in D. E. S. group without caponization was increased. upon 96.86% than initial period and A. C. T. H. group was 104.22% but other groups and all carponization groups were lighter than those of control group. 2) Weekly body gain of D. E. S. group without caponization was best showing the significance (102.69 g) and the group with caponization were lower than those groups without caponization. 3) Carcass yield was best in Testo. group without caponization (831.2 g) and the group with caponization were lower than the group without caponization. 4) Carcass rate was highest in A. C. T. H. group with caponization and (67.22%) lowest in Testo. group without caponization (63.37%), but any significance was not recognized. 2. The effects of caponizatitn and hormone treatments upon the coposition of meat and amino acids were; 1) Any significance was not recognized between treated and untreated group about change of moisture, crude protein, crude ash and glycogen contents in meat. 2) Fat co tent in muscle in the all treated groups were higher than that of control group. 3) Extracts of group without caponization were higher than those of groups with caponization. 4) Lysin contents were highest in D. E. S. group with caponization (11. 12/ 16.0 g N) and generelly Testo. group was lower compared with D. E. S. group. 5) Histidine and Arginine contents were higher in the groups with caponization than without caponization. 6) Aspartic acid content were higher in D. E. S. group and A. C. T. H. group without depend on caponization. 7) Treonine content was higher in Testo. group without caponization and in the group with caponization and without hormone treatment compared with those of control group without caponization. 8) Serine content was decreased in the group with caponization and increased by D. E. S. and A. C. T. H treatment groups and glutamic acid was also decreased in Testo. group with out caponization. 9) Cystine content was decreased by Testo. treatment and was not appeared in Testo. group without caponization. 10) Valine content was lower in control group with caponization but significance was not recognized between other groups and control group without caponization. 11) Glycine, Alanine, Methionine. Isoleucine, Leucine, Thyrosine and Phenylalanine contents were not so difference between hormone treated groups and control group without caponization. 3. The effects of caponization and hormone treatment upon the change of organs were: 1) The weight of all organs were heaviest in D. E. S. group without caponization (18.5g) and lightest in A. C. T. H. group without caponization (155. 3g) but no significance was recognized between hormone treatment groups. 2) Heart weight was heaviest in D. E. S. group without caponization (7.46 g) and lightest in Testo. group without caponization (5.95 g). 3) Liver weight was heaviest in D. E. S. group without caponization(32.89g) and lightest in hormone untreated group with caponization(29.66g). Significance was not recognized. 4) Spleen weight was heaivest in Testo. group with caponization (3.22 g) and lightest in D. E. S. group without caponization(2.00g) in contrast with the other groups. High significance was recognized among the groups (P<0.01). 5) Cloacal thymus weight was lightest in D. E. S. group with or without caponization compared with control group without caponization. High significance was recognized among the groups. 6) Muscle fat content was not appeared in A. C. T. H. group with caponization, but it was highly increased in D. E. S. group with or without caponization. 7) Testis weight was lightest in D. E. S. group (0.38g) compared with control group (2.66g). Significance was recognized among the groups. 8) Large intestine, small intestine and cecum weight and length were heavier and longer in D. E. S. group without caponization and control group without caponization was lighter than those of hormone treated groups. 4. The effects of caponization and hormone treatment upon histological change of testis and thyroid gland: 1) The histological change of testis was significantly appeared in D. E. S. group that seminifirous tubles was slowly atrophied, the funtion of spernatogenesis was ceased, spermatocyte was changed as degeneration by pyknosis and karyorrhexis and interstitial cell was also atrophied, but in Testo. and A. C. T. H. group were similar as control group. 2) The histological change of thyroid gland in Testo. and A. C. T. H. groups without caponization were similar to that of control group without caponization, but in D. E. S. group without caponization, was changed squamously. Thyroid gland of the groups with caponization, epithelium of was atrophied and changed squamously as degeneration by pyknosis and karyorrhexis and the function of thyroid gland was slowly ceased in colloid and in hormone treated group with caponization.

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National Survey of Sarcoidosis in Korea (유육종증 전국실태조사)

  • 대한결핵 및 호흡기학회 학술위원회
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.453-473
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    • 1992
  • Background: National survey was performed to estimate the incidence of sarcoidosis in Korea. The clinical data of confirmed cases were analysed for the practice of primary care physicians and pulmonary specialists. Methods: The period of study was from January 1991 to December 1992. Data were retrospectively collected by correspondence with physicians in departments of internal medicine, dermatology, ophthalmology and neurology of the hospitals having more than 100 beds using returning postcards. In confirmed and suspicious cases of sardoidosis, case record chart for clinical and laboratory findings were obtained in detail. Results: 1) Postcards were sent to 523 departments in 213 hospitals. Internal medicine composed 41%, dermatology 20%, ophthalmology 20% and neurology 19%. 2) Postcards were returned from 241 departments (replying rates was 48%). 3) There were 113 confirmed cases from 50 departments and 10 cases. The cases were composed from internal medicine (81%), dermatology (13%), ophthalmology (3%) and neurology (3%). 78 confirmed cases were analysed, which were composed from department of internal medicine (92%), dermatology (5%), and neurology (3%). 4) The time span for analysed cases was 1980 to 1992. one case was analysed in 1980 and the number gradually increased to 18 cases in 1991. 5) The majority of patients (84.4%) were in the age group of 20 to 49 years. 6) The ratio of male to female was 1 : 1.5. 7) The most common chief complains were respiratory symptoms, dermatologic symptoms, generalized discomforts, visual changes, arthralgia, abdominal pains, and swallowing difficulties in order. 16% of the patients were asymptomatic. 8) Mean duration between symptom onset and diagnosis was 2 months. 9) The most common symptoms were respiratory, general, dermatologic, ophthalmologic, neurologic and cardiac origin in order. 10) Hemoglobin, hematocrits and platelet were in normal range. 58% of the patients had lymphopenia measuring less than 30% of white cell count. The ratio of CD4 to CD8 lymphocytes was $1.73{\pm}1.16$ with range of 0.43 to 4.62. ESR was elevated in 43% of the cases. 11) Blood chemistry was normal in most cases. Serum angiotensin converting enzyme (S-ACE) was $66.8{\pm}58.6\;U/L$ with the range of 8.79 to 265 U /L. Proteinuria of more than 150 mg was found in 42. 9% of the patients. 12) Serum IgG was elevated in 43.5%, IgA in 45.5%, IgM in 59.1% and IgE in 46.7%. The levels of complement C3 and C4 were in the normal range. Anti-nuclear antibody was detected in 11% of the cases. Kweim test was performed in 3 cases, and in all cases the result was positive. 13) FVC was decreased in 17.3%, FEV1 in 11.5%, FEV1/FVC in 10%, TLC in 15.2%, and DLco in 64.7%. 14) PaO2 was decreased below 90 mmHg in 48.6% and PaCO2 was increased above 45 mmHg in 5.7%. 15) The percentage of macrophages in BAL fluid was $51.4{\pm}19.2%$, lymphocytes $44.4{\pm}21.1%$, and the ratio of CD4 to CD8 lymphocytes was $3.41{\pm}2.07$. 16) There was no difference in laboratory findings between male and female. 17) Hilar enlargement on chest PA was present in 87.9% (bilaterally in 78.8% and unilaterally in 9.1%). 18) According to Siltzbach's classification, stage 0 was 5%, stage 158.3%, stage 228.3%, and stage 38.3%. 19) Hilart enlargement on chest CT was present in 92.6% (bilaterally 76.4% and unilaterally in 16.2%). 20) HRCT was done in 16 cases. The most common findings were nodules, interlobular thickening, focal patchy infiltrations in order. Two cases was normal finding. 21) Other radiologic examinations showed bone change in one case and splenomegaly in two cases. 22) Gallium scan was done in 12 cases. Radioactivity was increased in hilar and mediastinal lymph nodes in 8 cases and in parenchyme in 2 cases. 23) The pathologic diagnosis was commonly performed by transbrochial lung biopsy (TBLB, 47.3%), skin and mediastinal lymph nodes biopsy (34.5%), peripheral lymph nodes biopsy (23.6%), open lung biopsy (18.2%) and bronchial biopsy in order. 24) The most common findings in pathology were non·caseating granuloma (100%), multi-nucleated giant cell (47.3%), hyalinized acellular scar (34.5%), reticulin fibrin network (20%), inclusion body (10.9%), necrosis (9.1%), and lymphangitic distribution of granuloma (1.8%) in order. Conclusion: Clinical, laboratory, radiologic and pathologic findings were summarized. This collected data will assist in finding a test for detection and staging of sarcoidosis in Korea in near future.

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