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Development and Validation of the Analytical Method for Oxytetracycline in Agricultural Products using QuEChERS and LC-MS/MS (QuEChERS법 및 LC-MS/MS를 이용한 농산물 중 Oxytetracycline의 잔류시험법 개발 및 검증)

  • Cho, Sung Min;Do, Jung-Ah;Lee, Han Sol;Park, Ji-Su;Shin, Hye-Sun;Jang, Dong Eun;Cho, Myong-Shik;Jung, ong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.227-234
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    • 2019
  • An analytical method was developed for the determination of oxytetracycline in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After the samples were extracted with methanol, the extracts were adjusted to pH 4 by formic acid and sodium chloride was added to remove water. Dispersive solid phase extraction (d-SPE) cleanup was carried out using $MgSO_4$ (anhydrous magnesium sulfate), PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed with LC-MS/MS using ESI (electrospray ionization) in positive ion MRM (multiple reaction monitoring) mode. The matrix-matched calibration curves were constructed using six levels ($0.001{\sim}0.25{\mu}g/mL$) and coefficient of determination ($r^2$) was above 0.99. Recovery results at three concentrations (LOQ, $10{\times}LOQ$, and $50{\times}LOQ$, n=5) were from 80.0 to 108.2% with relative standard deviations (RSDs) less than of 11.4%. For inter-laboratory validation, the average recovery was in the range of 83.5~103.2% and the coefficient of variation (CV) was below 14.1%. All results satisfied the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for oxytetracycline determination in agricultural commodities. This study could be useful for safety management of oxytetracycline residues in agricultural products.

Development of a Simultaneous Analytical Method for Determination of Insecticide Broflanilide and Its Metabolite Residues in Agricultural Products Using LC-MS/MS (LC-MS/MS를 이용한 농산물 중 살충제 Broflanilide 및 대사물질 동시시험법 개발)

  • Park, Ji-Su;Do, Jung-Ah;Lee, Han Sol;Park, Shin-min;Cho, Sung Min;Kim, Ji-Young;Shin, Hye-Sun;Jang, Dong Eun;Jung, Yong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.2
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    • pp.124-134
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    • 2019
  • An analytical method was developed for the determination of broflanilide and its metabolites in agricultural products. Sample preparation was conducted using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method and LC-MS/MS (liquid chromatograph-tandem mass spectrometer). The analytes were extracted with acetonitrile and cleaned up using d-SPE (dispersive solid phase extraction) sorbents such as anhydrous magnesium sulfate, primary secondary amine (PSA) and octadecyl ($C_{18}$). The limit of detection (LOD) and quantification (LOQ) were 0.004 and 0.01 mg/kg, respectively. The recovery results for broflanilide, DM-8007 and S(PFP-OH)-8007 ranged between 90.7 to 113.7%, 88.2 to 109.7% and 79.8 to 97.8% at different concentration levels (LOQ, 10LOQ, 50LOQ) with relative standard deviation (RSD) less than 8.8%. The inter-laboratory study recovery results for broflanilide and DM-8007 and S (PFP-OH)-8007 ranged between 86.3 to 109.1%, 87.8 to 109.7% and 78.8 to 102.1%, and RSD values were also below 21%. All values were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food and Drug Safety Evaluation guidelines (2016). Therefore, the proposed analytical method was accurate, effective and sensitive for broflanilide determination in agricultural commodities.

Transfer Learning using Multiple ConvNet Layers Activation Features with Principal Component Analysis for Image Classification (전이학습 기반 다중 컨볼류션 신경망 레이어의 활성화 특징과 주성분 분석을 이용한 이미지 분류 방법)

  • Byambajav, Batkhuu;Alikhanov, Jumabek;Fang, Yang;Ko, Seunghyun;Jo, Geun Sik
    • Journal of Intelligence and Information Systems
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    • v.24 no.1
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    • pp.205-225
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    • 2018
  • Convolutional Neural Network (ConvNet) is one class of the powerful Deep Neural Network that can analyze and learn hierarchies of visual features. Originally, first neural network (Neocognitron) was introduced in the 80s. At that time, the neural network was not broadly used in both industry and academic field by cause of large-scale dataset shortage and low computational power. However, after a few decades later in 2012, Krizhevsky made a breakthrough on ILSVRC-12 visual recognition competition using Convolutional Neural Network. That breakthrough revived people interest in the neural network. The success of Convolutional Neural Network is achieved with two main factors. First of them is the emergence of advanced hardware (GPUs) for sufficient parallel computation. Second is the availability of large-scale datasets such as ImageNet (ILSVRC) dataset for training. Unfortunately, many new domains are bottlenecked by these factors. For most domains, it is difficult and requires lots of effort to gather large-scale dataset to train a ConvNet. Moreover, even if we have a large-scale dataset, training ConvNet from scratch is required expensive resource and time-consuming. These two obstacles can be solved by using transfer learning. Transfer learning is a method for transferring the knowledge from a source domain to new domain. There are two major Transfer learning cases. First one is ConvNet as fixed feature extractor, and the second one is Fine-tune the ConvNet on a new dataset. In the first case, using pre-trained ConvNet (such as on ImageNet) to compute feed-forward activations of the image into the ConvNet and extract activation features from specific layers. In the second case, replacing and retraining the ConvNet classifier on the new dataset, then fine-tune the weights of the pre-trained network with the backpropagation. In this paper, we focus on using multiple ConvNet layers as a fixed feature extractor only. However, applying features with high dimensional complexity that is directly extracted from multiple ConvNet layers is still a challenging problem. We observe that features extracted from multiple ConvNet layers address the different characteristics of the image which means better representation could be obtained by finding the optimal combination of multiple ConvNet layers. Based on that observation, we propose to employ multiple ConvNet layer representations for transfer learning instead of a single ConvNet layer representation. Overall, our primary pipeline has three steps. Firstly, images from target task are given as input to ConvNet, then that image will be feed-forwarded into pre-trained AlexNet, and the activation features from three fully connected convolutional layers are extracted. Secondly, activation features of three ConvNet layers are concatenated to obtain multiple ConvNet layers representation because it will gain more information about an image. When three fully connected layer features concatenated, the occurring image representation would have 9192 (4096+4096+1000) dimension features. However, features extracted from multiple ConvNet layers are redundant and noisy since they are extracted from the same ConvNet. Thus, a third step, we will use Principal Component Analysis (PCA) to select salient features before the training phase. When salient features are obtained, the classifier can classify image more accurately, and the performance of transfer learning can be improved. To evaluate proposed method, experiments are conducted in three standard datasets (Caltech-256, VOC07, and SUN397) to compare multiple ConvNet layer representations against single ConvNet layer representation by using PCA for feature selection and dimension reduction. Our experiments demonstrated the importance of feature selection for multiple ConvNet layer representation. Moreover, our proposed approach achieved 75.6% accuracy compared to 73.9% accuracy achieved by FC7 layer on the Caltech-256 dataset, 73.1% accuracy compared to 69.2% accuracy achieved by FC8 layer on the VOC07 dataset, 52.2% accuracy compared to 48.7% accuracy achieved by FC7 layer on the SUN397 dataset. We also showed that our proposed approach achieved superior performance, 2.8%, 2.1% and 3.1% accuracy improvement on Caltech-256, VOC07, and SUN397 dataset respectively compare to existing work.

Diagnostic Usefulness of Serum Level of Cyfra 21-1, SCC Antigen and CEA in Lung Cancer (폐암에서 혈중 Cyfra 21-1, SCC 항원 및 CEA의 진단적 유용성)

  • Kim, Kyoung-Ah;Lee, Me-Hwa;Koh, Youn-Suck;Kim, Seon-Hee;Lim, Chae-Man;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Moon, Dae-Hyuk
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.6
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    • pp.846-854
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    • 1995
  • Background: Cytokeratin 19 is a subunit of cytokeratin intermediate filament expressed in simple epithelia such as respiratory epithelial cells and their malignant counterparts. An immunoradiometric assay is available to detect a fragment of the cytokeratin, referred to as Cyfra 21-1 in the serum. This study was conducted to evaluate the clinical utility of this new marker in the diagnosis of lung cancer compared with established markers of squamous cell carcinoma antigen (SCC Ag) and carcino-embryonic antigen(CEA). In addition, we compared the diagnostic sensitivity and specificity of Cyfra 21-1 with those of SCC Ag in squamous cell carcinoma of the lung. We also measured the level of Cyfra 21-1 in the different stages of squamous cell carcinoma of the lung. Method: We measured Cyfra 21-1(ELSA-CYFRA 21-1), SCC Ag(ABBOTT SCC RIABEAD) and CEA(ELSA2-CEA) in 79 patients with primary lung cancer and in 78 persons as a comparison group including 32 patients with pulmonary tuberculosis, 23 patients with benign lung disease and 23 cases with healthy individual. Cyfra 21-1 is measured by a solid-phase immunoradiometric assay(CIS Bio International, France) based on the two-site sandwich method. SCC Ag is measured by a radioimmunoassay(Abbott Laboratories, USA). CEA is measured by a immunoradiometric assay(CIS Bio International, France). All data were expressed as the mean$\pm$standard deviation. Results: 1) The mean value of Cyfra 21-1 was $18.38{\pm}3.65\;ng/mL$ in the lung cancer and $1.l6{\pm}0.53\;ng/mL$ in the comparison group(p<0.0001). SCC Ag was $3.53{\pm}6.06\;ng/mL$ in the lung cancer and $1.19{\pm}0.5\;ng/mL$ in the comparison group(p<0.01). CEA was $35.03{\pm}13.9\;ng/mL$ in the lung cancer and $2.89{\pm}1.01\;ng/mL$ in the comparison group(p<0.0001). 2) Cyfra 21-1 level in squamous cell carcinoma($31.52{\pm}40.13\;ng/mL$) was higher than that in adenocarcinoma($2.41{\pm}1.34\;ng/mL$)(p<0.0001) and small cell carcinoma($2.15{\pm}2.05\;ng/mL$)(p=0.007). SCC Ag level in squamous cell carcinoma($5.1{\pm}7.68\;ng/mL$) was higher than that in adenocarcinoma($1.36{\pm}0.69\;ng/mL$)(p=0.009) and small cell carcinoma($1.1{\pm}0.24\;ng/mL$) (p=0.024). 3) The level of Cyfra 21-1 was not correlated with the progression of stage in squamous cell carcinoma of the lung. 4) Using the cut-off value of 3.3ng/mL, the diagnostic sensitivity of Cyfra 21-1 was 83% in squamous cell carcinoma, 22% in adenocarcinoma and 17% in small cell carcinoma. The sensitivity of SCC Ag and CEA were 39% and 20%, respectively in squamous cell carcinoma, 11% and 39% in adenocarcinoma, and 0% and 33% in small cell carcinoma. 5) Comparison of the receiver operating characteristics curves(ROC curve) for Cyfra 21-1, SCC Ag and CEA revealed that Cyfra 21-1 showed highest diagnostic sensitivity among them in the diagnosis of lung cancer. Conclusion: Cyfra 21-1 is thought to be a better tumor marker for the diagnosis of lung cancer than SCC Ag and CEA, especially in squamous cell carcinoma of the lung.

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