• Title/Summary/Keyword: molecular method

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Evaluation of Renal Pathophysiological Processes Induced by an Iodinated Contrast Agent in a Diabetic Rabbit Model Using Intravoxel Incoherent Motion and Blood Oxygenation Level-Dependent Magnetic Resonance Imaging

  • Yongfang Wang;Xin Zhang;Bin Wang;Yang Xie;Yi Wang;Xuan Jiang;Rongjia Wang;Ke Ren
    • Korean Journal of Radiology
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    • v.20 no.5
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    • pp.830-843
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    • 2019
  • Objective: To examine the potential of intravoxel incoherent motion (IVIM) and blood oxygen level-dependent (BOLD) magnetic resonance imaging for detecting renal changes after iodinated contrast-induced acute kidney injury (CI-AKI) development in a diabetic rabbit model. Materials and Methods: Sixty-two rabbits were randomized into 2 groups: diabetic rabbits with the contrast agent (DCA) and healthy rabbits with the contrast agent (NCA). In each group, 6 rabbits underwent IVIM and BOLD imaging at 1 hour, 1 day, 2 days, 3 days, and 4 days after an iohexol injection while 5 rabbits were selected to undergo blood and histological examinations at these specific time points. Iohexol was administrated at a dose of 2.5 g I/kg of body weight. Further, the apparent transverse relaxation rate (R2*), average pure molecular diffusion coefficient (D), pseudo-diffusion coefficient (D*), perfusion fraction (f) were calculated. Results: The D and f values of the renal cortex (CO) and outer medulla (OM) were significantly decreased compared to baseline values in the 2 groups 1 day after the iohexol injection (p < 0.05). A marked reduction in the D* values for both the CO and OM was also observed after 1 hour in each group (p < 0.05). In the OM, a persistent elevation of the R2* was detected for 4 days in the DCA group (p < 0.05). Histopathological changes were prominent, and the pathological features of CI-AKI aggravated in the DCA group until day 4. The D, f, and R2* values significantly correlated with the histological damage scores, hypoxia-inducible transcription factor-1α expression scores, and serum creatinine levels. Conclusion: A combination of IVIM and BOLD imaging may serve as a noninvasive method for detecting and monitoring CI-AKI in the early stages in the diabetic kidney.

A Study on KASP Analysis Based on SNP to Rapidly Identify Caviar-Producing Sturgeon Species (캐비어를 생산하는 철갑상어의 신속 종판별을 위한 SNP 기반 KASP 분석에 관한 연구)

  • Sun Hee Lee;Bo Reum Park;Hyung Il Kim;Sooyeul Cho;Kyung-Hun Son
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.209-220
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    • 2024
  • Cavier is graded as Beluga, Osetra, and Sevruga based on the species of sturgeon (Acipenser sinensis). In this study, we developed an analytical method for determining the grade of black caviar using DNA barcodes and KASP markers. To identify the sturgeon species, ten black caviar samples were collected, and a reference DNA barcode library was developed using five genes (namely, 16S ribosomal RNA, cytochrome b, cytochrome c oxidase subunit I, cytochrome c oxidase subunit II, and NADH dehydrogenase subunit 5 genes). To develop the KASP markers, we selected 11 markers that could distinguish between the five grades of black caviar. As a result, specific markers for each of the targeted caviars were clustered into FAM-positive sections. DNA barcoding and the KASP assay revealed that one Beluga, six Osetra, and three Sevruga were identified among the ten caviar samples. Moreover, we found that the sturgeon species were mislabeled in two products. Here, we aimed to develop a KASP assay based on SNP that allows rapid and easy identification of caviar grade. These methods are expected to contribute to preventing the distribution of illegal products.

Development of Geometric Calibration Method for Triple Head Pinhole SPECT System (삼중헤드 SPECT에서 기하학적 보정 기법의 개발)

  • Kim, Joong-Hyun;Lee, Jae-Sung;Lee, Won-Woo;Park, So-Yeon;Son, Ji-Yeon;Kim, Yu-Kyeong;Kim, Sang-Eun;Lee, Dong-Soo
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.1
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    • pp.61-69
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    • 2008
  • Purpose: Micro-pinhole SPECT system with conventional multiple-head gamma cameras has the advantage of high magnification factor for imaging of rodents. However, several geometric factors should be calibrated to obtain the SPECT image with good image quality. We developed a simplified geometric calibration method for rotating triple-head pinhole SPECT system and assessed the effects of the calibration using several phantom and rodent imaging studies. Materials and Methods: Trionix Triad XLT9 triple-head SPECT scanner with 1.0 mm pinhole apertures were used for the experiments. Approximately centered point source was scanned to track the angle-dependent positioning errors. The centroid of point source was determined by the center of mass calculation. Axially departed two point sources were scanned to calibrate radius of rotation from pinhole to center of rotation. To verify the improvements by the geometric calibration, we compared the spatial resolution of the reconstructed image of Tc-99m point source with and without the calibration. SPECT image of micro performance phantom with hot rod inserts was acquired and several animal imaging studies were performed. Results: Exact sphere shape of the point source was obtained by applying the calibration and axial resolution was improved. Lesion detectibility and image quality was also much improved by the calibration in the phantom and animal studies. Conclusion: Serious degradation of micro-pinhole SPECT images due to the geometric errors could be corrected using a simplified calibration method using only one or two point sources.

Development of an Efficient Method of Screening for Watermelon Plants Resistant to Fusarium oxysporum f. sp. niveum (수박 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Jo, Eun Ju;Lee, Ji Hyun;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.409-419
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    • 2015
  • This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.

Establishment of Pre-Harvest Residue Limit (PHRL) of Fungicides Azoxystrobin and Difenoconazole on Prunus mume fruits (매실 중 살균제 azoxystrobin과 difenoconazole의 생산단계 잔류허용기준 설정)

  • Lee, Dong Yeol;Kim, Yeong Jin;Park, Min Ho;Lee, Seung Hwa;Kim, Sang Gon;Kang, Nam Jun;Kang, Kyu Young
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.307-313
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    • 2013
  • This study was carried out to investigate the residual characteristics of fungicide azoxystrobin and difenoconazole in Prunus mume fruits, and establish pre-harvest residue limits (PHRL) based on dissipation and biological half-lives of fungicide residues. The fungicides were sprayed onto the crop at recommended dosage once and 3 times in 7 days interval, respectively. The samples were harvested at 0, 1, 2, 4, 6, 8, 10, 12 and 14 days after treatment. These residual pesticides were extracted with QuEChERS method, clean-up with $NH_2$ SPE cartridge, and residues were analyzed by HPLC/DAD and GLC/ECD, respectively. Method quantitative limits (MQL) of azoxystrobin were 0.03 mg $kg^{-1}$ and of difenoconazole were 0.006 mg $kg^{-1}$. Average recovery were $93.2{\pm}2.49%$, $85.5{\pm}1.97%$ for azoxystrobin at fortification levels at 0.3 and 1.5 mg $kg^{-1}$, and $100.8{\pm}6.74%$, $87.6{\pm}9.92%$ for difenoconazole at fortification levels at 0.06 and 0.3 mg $kg^{-1}$, respectively. The biological half-lives of azoxystrobin were 5.9 and 5.2 days at recommended dosage once and 3 times in 7 days interval, respectively. The biological half-lives of difenoconazole were 9.3 and 8.0 days at recommended dosage once and 3 times in 7 days interval, respectively. The PHRL of azoxystrobin and difenoconazole were recommended as 5.32 and 1.64 mg $kg^{-1}$ for 10 days before harvest, respectively.

Optimization of Analytical Methods for Octacosanol in Related Health-functional Foods with GC-MS (GC-MS를 이용한 건강기능식품 중 옥타코사놀 분석법 개발 연구)

  • Lee, Jin Hee;Oh, Mi Hyune;Lee, Kyung Jin;Kim, Yang Sun;Keum, Eun Hee;Park, Ji Eun;Cho, Mee Hyun;Seong, Min Hye;Kim, Sang A;Kim, Mee hye
    • Journal of Food Hygiene and Safety
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    • v.33 no.4
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    • pp.266-271
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    • 2018
  • The Ministry of Food and Drug Safety (MFDS) is amending its test methods for health-functional foods (dietary food supplements) to establish regulatory standards and specifications in Korea. In this regard, we continue our research on developing analytical methods for the items. Octacosanol is the major component of polycosanol and is a high-molecular-mass primary fatty alcohol, obtained from sugar cane wax. Previous researchers have shown that octacosanol can lower cholesterol and has antiaggregatory properties, cytoprotective uses, and ergogenic properties for human health. Recently, octacosanol products have been actively introduced into the domestic market because of their functional biological activity. We have developed a sensitive and selective test method for octacosanol that the TMS derivatives by means of gas-chromatographic-tandem mass spectrometry (GC-MS). The trimethylsilyl ether derivative of the target analyte showed excellent chromatographic properties. The procedure was validated in the range of $12.5{\sim}200{\mu}g/L$. Standard calibration curves presented linearity with the correlation coefficient ($r^2$) > 0.999, and the limits of detection (LOD) and limits of quantitation (LOQ) were $4.5{\mu}g/L$ and $13.8{\mu}g/L$, respectively. The high recoveries (92.5 to 108.8%) and precision (1.8 to 2.4%) obtained are in accordance with the established validation criteria. Our research can provide scientific evidence to amend the octacosanol test method for the Health-Functional Food Code.

Preparation of Soluble Dietary Fiber from Oak Wood (Quercus Mongolica) and Its Physiological Function in Rat Fed High Cholesterol Diets (참나무 (Quercus Mongolica)로부터 수용성 식이섬유소의 제조 및 기능성 검증)

  • 채영미;임부국;이종윤;김영희;이순재
    • Journal of Nutrition and Health
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    • v.36 no.1
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    • pp.9-17
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    • 2003
  • The preparation method of a soluble dietary fiber from oak wood (Quercus mongolica) and the effect of the soluble dietary fiber on physiological function in rat fed high cholesterol diets was investigated. The best condition for steam explosion method was 25 kgf/㎤ pressure for 6 min. The exploded samples were delignified by the filtration treatment with 1% NaOH for several times, which is the best condition. The enzymatic hydrolysis of Cellusoft cellulase was more effective than Onozuka R-10 cellulase. The manufactured soluble dietary fiber was assayed using gel permeation chromatography (GPC) and it was dissolved in water. Average molecular weight distribution of manufactured soluble dietary fiber was about 348-1,200 and it was assumed the oligomer form fraction. In order to compare the manufactured soluble dietary fiber with commercial soluble dietary fiber (pectin) on the physiological function, Sprague-Dawley male rats weighing 100$\pm$10 g were randomly assigned to one normal diet and five high cholesterol diet containing 1% cholesterol. The high cholesterol diet groups were classified to fiber free diet (FF group), 5% pectin (5P group), 10% pectin (l0P group), 5% manufactured soluble dietary fiber (5M group) and 10% manufactured soluble dietary fiber (10M group). Body weight gains in all soluble dietary fiber groups were lower than FF group. Food intakes were increased in all soluble dietary fiber groups than that of FF group. Food efficiency ratio (FER) was significantly decreased in all soluble dietary fiber groups than that of the FF group, and it was especially was highest in 10% supplemented soluble dietary fiber group. The weight of liver of the soluble dietary fiber supplemented groups were lower than those of the FF group, but weights of cecum and small intestine of all supplemented soluble dietary fiber groups were significantly increased, compared with that of FF group. The weights and water contents in feces were significantly increased by the soluble dietary fiber. The activity of the glutamic oxaloacetic transaminase in soluble dietary fiber groups were significantly decreased than those of FF group. The hepatic glutathione S-transferase activity in all soluble dietary fiber supplemented groups were higher than that of FF group. The physiological effects of the manufactured soluble dietary fiber are the same as the commercial soluble dietary fiber (pectin). The preparation method of the soluble dietary fiber from the oak chips suited to its purpose. (Korean J Nutrition 36(1) : 9~17, 2003)

Determination of Appropriate Sampling Frequency and Time of Multiple Blood Sampling Dual Exponential Method with $^{99m}Tc$-DTPA for Calculating GFR (사구체여과율 계산을 위한 $^{99m}Tc$-DTPA를 이용한 다중 채혈 이중지수법의 적정 채혈 횟수 및 시간의 선정)

  • Kim, Chung-Ho;O, Joo-Hyun;Chung, Yong-An;Yoo, Ie-Ryung;Sohn, Hyung-Sun;Kim, Sung-Hoon;Chung, Soo-Kyo;Lee, Hyoung-Koo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.1
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    • pp.33-39
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    • 2006
  • Purpose: To determine appropriate sampling frequency and time of multiple blood sampling dual exponential method with $^{99m}Tc$-DTPA for calculating glomerular filtration rate (GFR). Materials & Methods: Thirty four patients were included in this study. Three mCi of $^{99m}Tc$-DTPA was intravenously injected and blood sampling at 9 different times, 5ml each, were done. Using the radioactivity of serum, measured by gamma counter, the GFR was calculated using dual exponential method and corrected with the body surface area. Using spontaneously chosen 2 data points of serum radioactivity, 15 collections of 2-sample GFR were calculated. And 10 collections of 3-sample GFR and 12 collections of 4-sample GFR were also calculated. Using the 9-sample GFR as a reference value, degree of agreement was analyzed with Kendall's $\tau$ correlation coefficients, mean difference and standard deviation. Results: Although some of the 2-sample GFR showed high correlation coefficient, over or underestimation had evolved as the renal function change. The 10-120-240 min 3-sample GFR showed a high correlation coefficient (${\tau}=0.93$), minimal difference ($Mean{\pm}SD=-1.784{\pm}3.972$), and no over or underestimation as the renal function changed. The 4-sample GFR showed no better accuracy than the 3-sample GFR. Conclusions: In the wide spectrum of renal function, the 10-120-240 min 3-sample GFC could be the best choice for estimating the patients' renal function.

Occurrence of Meloidogyne incognita Infecting Resistant Cultivars and Development of an Efficient Screening Method for Resistant Tomato to the Mi-virulent Nematode (뿌리혹선충 저항성 토마토를 감염하는 Meloidogyne incognita의 발생 및 이 선충을 이용한 효율적인 저항성 검정법 확립)

  • Hwang, Sung Min;Park, Myung Soo;Kim, Jin-Cheol;Jang, Kyoung Soo;Choi, Yong Ho;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.217-226
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    • 2014
  • Root-knot symptoms were found on a commercial tomato cultivar carrying Mi, a resistance gene to root-knot nematodes including Meloidogyne incognita, M. arenaria, and M. javanica in 2012 at Buyeo, Chungnam Province in Korea. The isolate was identified as M. incognita based on molecular analyses using two species-specific primer sets. Pathogenicity of the isolate on one susceptible and three resistant tomato cultivars to the root-knot nematodes was tested. The nematode isolate showed strong pathogenicity on all the tested cultivars at all tested incubation temperatures. In addition, resistance degree of 33 commercial tomato cultivars, 8 susceptible and 25 resistant cultivars to root-knot nematodes, was also tested. Plants were determined as resistant when they suppressed the nematode reproduction. All the cultivars demonstrated strong susceptibility to the nematode regardless of resistance of the tomato cultivars. To our knowledge, this is the first report on the occurrence of Mi infecting M. incognita isolate in Korea. On the other hand, to construct an efficient screening method for selecting resistant breeding source to the nematode isolate, root-knot development of M. incognita on four tomato cultivars according to several conditions such as inoculum concentration, plant growth stage, and incubation period after transplant was investigated. Reproduction of the nematode on all the tested cultivars according to inoculum concentration increased in a dose-dependent manner. Except for inoculum concentration, there was no significant difference in reproduction level of the cultivars according to the other tested conditions. On the basis of the results, we suggest an efficient screening method for new resistant tomato to the nematode isolate.

Laboratory-scale fruiting body formation of Pleurotus ostreatus using the petri dish culture (느타리의 기내 자실체 형성 및 그 유도조건에 관한 연구)

  • Joh, Joong-Ho;Chu, Kyo-Sun;Kim, Beom-Gi;Kong, Won-Sik;Yoo, Young-Bok;Lee, Seung-Jae;Cho, Bong-Gum;Lee, Chang-Soo
    • Journal of Mushroom
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    • v.2 no.1
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    • pp.15-20
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    • 2004
  • Pleurotus ostreatus, the oyster mushroom, is one of the most widely cultivated and important edible mushrooms in the world. In order to study the developmental process of P. ostreatus and its regulatory mechanism, a new culturing method needs to be established for inducing the fruiting body and sporulation in the laboratory. In this study, we have examined whether the fruiting body of P. ostreatus can be formed on the plastic petri dish which are commonly used for cell culture in the laboratory. The strain was cultured on $60{\times}15mm$ plastic petri dish with potato dextrose agar media at $28^{\circ}C$ for mycelial growth and then at $18^{\circ}C$ for the formation of primordia and fruiting bodies within plant growth chamber. The development of primordia into fruiting bodies was achieved on cultured dishes under air ventilation. At the primordia stage, the normal formation of fruiting body was blocked by sealing the plastic dish with parafilm. The periods requiring for the formation of primordia and fruiting bodies were examined on the dish culture. About 96% and 76% of cultured samples formed primordia and fruiting bodies under the optimal conditions during ten weeks of culture, respectively. These culturing periods, however, were changed by the mechanical injury treatment to mycelia. As other factors affecting the fruiting body formation, the effects of light and cold shock have been tested. No fruiting formation was observed on the cultured dishes under the dark. The cold shock treatment by storing cultured dishes for one day at $4^{\circ}C$ did not have any significant effects in the fruiting body formation. Spores of fruiting bodies acquired from the petri dishes could be germinated on culture media at $28^{\circ}C$. These results suggest that the fruiting bodies of P. ostreatus can be formed on the experimental petri dish and this dish-culturing method is useful for understanding of the developmental process of P. ostreatus in the laboratory. Furthermore, the dish-culturing method is able to shorten the life cycle of P. ostreatus without requiring large area and expensive device.

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