This study was conducted to reset the maximum residue limit (MRL) for didecyldimethylammonium chloride (DDAC) in broiler chickens. The disinfectant containing DDAC (10%, w/w) was diluted 160 times and evenly sprayed on the bodies of twenty-four broiler chickens at a rate of 15 mL per day per bird for 7 days. After the disinfectant treatment, tissue samples were collected from six broiler chickens at 0.25, 1, 3 and 5 days, respectively. Residual DDAC concentrations in poultry tissues were determined using LC-MS/MS. Correlation coefficient (0.99 >), the limits quantification (LOQ, 2.0~10.0 μg/kg), recoveries (86.9~118.6%), and coefficient of variations (<19.98%) were satisfied the validation criteria of Korean Ministry of Food and Drug Safety. In all tissues except for liver, DDAC was detected more than LOQ at 5 days after the disinfectant treatment. In liver tissues, DDAC was detected more than LOQ at 3 days after treatment. According to the European Medicines Agency’s guideline on determination of withdrawal periods, withdrawal period of DDAC in poultry tissues was established to 26 days. In conclusion, the developed analytical method is sensitive and reliable for detecting DDAC in poultry tissues. When DDAC disinfectant is sprayed on a poultry house in the presence of broiler chickens, it is necessary to keep the disinfectant from contacting the body of the livestock.
This study investigated ethopabate (EPB) residues in edible tissues of broiler chickens given in drinking water and established the withdrawal time (WT) of EPB in poultry tissues. Twenty-four healthy Ross broiler chickens were orally administered with EPB at the concentration of 3.8 mg/L for 14 days (EPB-1, n=24) and 15.2 mg/L for 7 days (EPB-2, n=24) through drinking water, respectively. After the drug treatment, tissue samples were collected from six broiler chickens at 0, 1, 3, and 5 days, respectively. EPB residue concentrations in poultry tissues were determined using LC-MS/MS. Correlation coefficient values ranged from 0.9980 to 0.9998, and the limits of detection and quantification (LOQ) were 0.03~0.09 and 0.1~0.3 μg/kg, respectively. Mean recoveries in muscle, liver, kidney and skin/fat tissues were 95.9~109.8, 108.7~115.3, 89.9~96.6 and 86.7~96.8%, respectively, and coefficient of variations were less than 17.11%. At the end of the drug-administration period (0 day), EPB was detected at levels under the LOQ in all tissues from both the EPB-1 and EPB-2 groups. According to the results of EPB residue in Ross broiler tissues, withdrawal periods of both EPB-1 and EPB-2 in poultry tissues were established to 0 day. In conclusion, the developed analytical method is suitable for the detection of EPB in poultry tissues, and the estimated WT of EPB in poultry tissues will contribute to ensuring the safety of Ross broiler chickens.
From 2020, Korean Animal and Plant Quarantine Agency has reset the withdrawal time (WT) for veterinary drugs typically used in livestock in preparation for the introduction of positive list system (PLS) program in 2024. This study was conducted to reset the MRL for amprolium (APL) in broiler chickens as a part of PLS program introduction. Forty-eight healthy Ross broiler chickens were orally administered with APL at the concentration of 60 mg/L (APL-1, n=24) for 14 days and 240 mg/L (APL-2, n=24) for 7 days through drinking water, respectively. After the drug treatment, tissue samples were collected from six broiler chickens at 0, 1, 3 and 5 days, respectively. Residual APL concentrations in poultry tissues were determined using LC-MS/MS. Correlation coefficient (0.99 >), the limits quantification (LOQ, 0.3~5.0 μg/kg), recoveries (81.5~112.4%), and coefficient of variations (<15.5%) were satisfied the validation criteria of Korean Ministry of Food and Drug Safety. In APL-1, APL in all tissues except for kidney was detected less than LOQ at 3 days after drug treatment. In APL-2, APL in liver and kidney was detected more than LOQ at 5 days after treatment. According to the European Medicines Agency’s guideline on determination of withdrawal periods, withdrawal periods of APL-1 and APL-2 in poultry tissues were established to 3 and 2 days, respectively. In conclusion, the developed analytical method is sensitive and reliable for detecting APL in poultry tissues. The estimated WT of APL in poultry tissues is longer than the current WT recommendation of 2 days for APL in broiler chickens.
본 연구는 소의 가식부위(근육, 신장, 간장, 지방) 중에 서 세팔렉신을 효과적으로 정량분석하기 위한 LC-MS/MS 법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준 용액을 이용하여 검량성을 작성한 결과, r2 > 0.999 이상의 직선성을 나타내었으며, 세팔렉신에 대한 검출한계와 정량한계는 각각 2~10과 6~30 μg/kg으로 나타났다. 또한, 회 수율은 83.9~106.8%로 나타났으며, 상대표준편차는 2.3~ 14.8%로 나타나 정확성이 우수하였다. 이는 식품의약품안 전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LCMS/ MS법은 향후 소의 가식부위 중 세팔렉신을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.
본 연구는 우유 중에서 덱사메타손을 효과적으로 정량 분석하기 위한 LC-MS/MS법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출 한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준용액을 이용하여 검량성을 작성한 결과, r2 > 0.999 이상의 직선성을 확인하였고, 덱사 메타손에 대한 검출한계와 정량한계는 각각 0.15와 0.5 ng/ mL이었다. 또한, 회수율은 98.9-109.6%로 나타났으며, 상대표준편차는 1.7-4.4%로 나타나 정확성이 우수하였으며, 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 우유 중 덱사메타손을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.
Schwann cells play an important role in peripheral nerve regeneration. Upon nerve injury, Schwann cells are activated and produce various proinflammatory mediators including IL-6, LIF and MCP-1, which result in the recruitment of macrophages and phagocytosis of myelin debris. However, it is unclear how the nerve injury induces Schwann cell activation. Recently, it was reported that necrotic cells induce immune cell activation via toll-like receptors (TLRs). This suggests that the TLRs expressed on Schwann cells may recognize nerve damage by binding to the endogenous ligands secreted by the damaged nerve, thereby inducing Schwann cell activation. To explore the possibility, we stimulated iSC, a rat Schwann cell line, with damaged neuronal cell extracts (DNCE). The stimulation of iSC with DNCE induced the expression of various inflammatory mediators including IL-6, LIF, MCP-1 and iNOS. Studies on the signaling pathway indicate that NF-xB, p38 and JNK activation are required for the DNCE-induced inflammatory gene expression. Furthermore, treatment of either anti-TLR3 neutralizing antibody or ribonuclease inhibited the DNCE-induced proinflammatory gene expression in iSC. In summary, these results suggest that damaged neuronal cells induce inflammatory Schwann cell activation via TLR3, which might be involved in the Wallerian degeneration after a peripheral nerve injury.
The mesencephalic trigeminal nucleus (Mes V) contains cell bodies of primary afferent sensory neurons that relay proprioceptive information from the periodontium and masticatory muscles and function as typical sensory neurons or potentially as integrative interneurons. In the present study, we studied these two potential functions using combined experimental approaches of retrograde labeling and whole cell patch clamp recording. Mes V neurons that presumably originate from periodontal nerve fibers in subsets of Mes V nucleus were identified by retrograde labeling with a fluorescent dye, DiI, which was applied onto inferior alveolar nerve. These cells were elliptical perikarya shaped cells about 40μmin diameter. In these neurons, we measured high voltage-activated calcium channel (HVACC) currents GABAв agonist, baclofen, inhibited calcium currents, and the HVACC currents inhibition by baclofen was voltage-dependent, exhibited prepulse facilitation, indicating that it was mediated by Gi/Go protein. Taken together, our results demonstrate that Mes V neurons not only have cell bodies originating from periodontium, but also receive synaptic inputs including GABAergic neurons suggesting that Mes V neurons function as both primary sensory neurons and integrative interneurons.
Background : Korean mountain ginseng(Panax ginseng C.A. Meyer) are difficult to clinically apply because of its scarcity and high cost. Advances in plant biotechnology have made it possible to produce mountain ginseng extracts on a large scale using adventitious root cultures in bio-reactors. This study investigated the variations of ginsenoside compounds composition and biological activities of wild ginseng adventitious roots by fermentation process. Methods and Results : Wild ginseng adventitious roots with five days fermentation using four strain of bacteria(Leuconostoc mesenteroides(KACC 15744), Bacillus circulans(KACC 15822), Bacillus licheniformis(KACC 15823), Bacillus subtilis subsp. inaquosorum(KACC 17047)). Ginsenoside contents was analysed by using HPLC. To examine the antioxidant activity associated with biological functions, radical scavenging analyses DPPH, ABTS and SOD-like activity analyses were conducted. The total phenolic and flavonoid contents were evaluated to determine the antioxidant activity increment. The result showed increased total ginsenoside contents by fermentation process. In particular, B. licheniformis showed the highest ginsenoside contents. Regarding ginseng fermented with B. licheniformis, values of 70.6 ± 1.4%, 44.3 ± 1.7%, and 88.4 ± 1.3% were measured using DPPH, ABTS, and SOD-like antioxdiant activity analyses, respectively. The total phenolic contents in ginseng fermented with B. licheniformis was 184.5 ± 0.9 ㎍·GAE/㎖, and the total flavonoid contents was 108.5 ± 1.8 ㎍·QE/㎖ in ginseng fermented with L. mesenteroides. Conclusion : The high activity of β-glucosidase was selected bacteria. The four types of lactic acid bacteria examined, the use of B. licheniformis to ferment ginseng resulted in greatest increase in biological activities and ginsenoside contents.