• 한국축산식품학회지
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• 제41권6호
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• pp.1022-1035
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• 2021

This study estimated the effect of exposure to propiconazole through implementation and residues in finishing pigs. We analyzed the expression of fibrosis-related genes and performed histological analysis of the blood, liver, kidney, muscle, ileum, and fat tissues. The animals were exposed for 28 d to different concentrations of propiconazole (0.09, 0.44, 0.88, 4.41, and 8.82 mg/kg bw/d). Quantitative, gene expression, and histological analyses in tissues were performed using liquid chromatography mass spectrometry, real-time PCR, and Masson's trichrome staining, respectively. Final body weight did not differ among groups. However, genes involved in fibrosis were significantly differentially regulated in response to propiconazole concentrations. Glucose, alanine aminotransferase, and total bilirubin levels were significantly increased compared with those in the control group, while alkaline phosphatase level was decreased (p<0.05) after exposure to propiconazole. The residue limits of propiconazole were increased in the finishing phase at 4.41 and 8.82 mg/kg bw/d. The liver, kidney, and ileum showed blue staining after propiconazole treatment, confirmed by Masson's trichrome staining. In conclusion, these findings suggest that propiconazole exposure disturbs the expression of fibrosis-related genes. This study on dietary propiconazole in pigs can provide a basis for determining maximum residue limits and a better understanding of metabolism in pigs and meat products.

• 대한수의학회지
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• 제60권4호
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• pp.209-213
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• 2020

This study examined the residue of dl-methylephedrine hydrochloride (MEP) on the muscle of pigs administered orally with MEP 12 g/ton feed for seven consecutive days. Twenty healthy cross swine were administered MEP. Four treated animals were selected arbitrarily to be sacrificed at 1, 2, 3, 4, and 5 days after treatment. MEP residue concentrations in the muscle were determined by liquid chromatography coupled with tandem mass spectrometry. The drug was extracted from muscle samples using 10 mM ammonium formate in acetonitrile followed by clean-up with n-hexane. The analyte was separated on an XBridgeTM hydrophilic interaction liquid chromatography column using 10 mM ammonium formate in deionized distilled water and acetonitrile. The correlation coefficient (R2) of the calibration curve was 0.9974, and the limits of detection and quantification were 0.05 and 0.15 ㎍/kg, respectively. The recoveries at three spiking levels were 94.5-101.2%, and the relative Standard Deviations was less than 4.06%. In the MEP-treated group, MEP residues on one day post-treatment were below the maximum residue limit in the muscle. The developed method is sensitive and reliable for the detection of MEP in porcine muscle tissues. Furthermore, it exhibits low quantification limits for animal-derived food products destined for human consumption.

• 대한수의학회지
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• 제63권3호
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• pp.23.1-23.9
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• 2023

This study was conducted to investigate tiamulin (TML) residues in the edible tissues of orally dosed broiler chickens and to re-establish the withdrawal time (WT). Thirty-six healthy Ross broiler chickens were administered 0.5 (TML-1) and 2.5 kg (TML-2) per ton feed, respectively, of the drug containing TML 78 g/kg for 10 days. Twenty-four tissue samples were collected from 6 chickens in each of the TML-1 and TML-2 groups on 0, 1, 3, and 5 days after drug administration, respectively. The residual concentrations of TML were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The correlation coefficient of the calibration curves was 0.9978 to 0.9998, and the limits of detection and the limits of quantification (LOQ) were in the range of 0.03 to 0.06, and 0.1 to 0.2 ㎍/kg, respectively. Recoveries ranged between 89.0% to 116.7%, and the coefficients of variation were less than 13.9%. After the drug administration, TML in the TML-1 and TML-2 groups was detected above the LOQ in 1 and 6 samples of liver, respectively, at day 0, and in 1 liver sample from both groups on day one. At 3 days after administration, TML was detected below the LOQ in all samples of TML-1 and TML-2. The calculated WT of TML in both TML-1 and TML-2 using the WT calculation program WT 1.4 was 0 days. In conclusion, the developed analytical method is suitable for detection, and the calculated WT of TML in poultry edible tissues is shorter than the current recommended WT of 7 days for TML in broiler chickens.

• 한국수산과학회지
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• 제57권1호
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• pp.41-52
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• 2024

Serotransferrin cDNA from the Siberian sturgeon Acipenser baerii was isolated and its expression patterns during early life intervals were characterized. It contained an ORF encoding a 708-aa-long polypeptide, including a 19-aa signal peptide. Bioinfomatic analysis and 3D modeling indicated a typical bi-lobal structure with conserved iron-coordinating residues. During embryonic development, the potential transition of maternally provisioned transcripts to zygotically de novo transcribed ones occurred around blastula stage. The transferrin mRNA levels peaked at stages responsible for pronephros, heart and erythropoietic component differentiation. After hatching, the transferrin mRNA expression gradually increased at early ontogenic phases (0 to 3 DPH) corresponding to the periods in which prolarvae exhibited increased blood circulation and liver differentiation. The expression decreased at subsequent stages in which prolarvae exhibited benthic movement. The tissue distribution assay indicated liver-predominant expression at fingerling stage. From the microinjection-based challenge with Aeromonas hydrophila at day-0 and day-7, the transcriptional response was modulated toward upregulation, in which the amounts induced at 6, 12 and 24 HPI were greater in prolarvae injected at day-7 than at day-0. Therefore, transferrin plays important roles in both early development and host protective responses to pathogens in the Siberian sturgeon.

• BMB Reports
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• 제40권5호
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• pp.715-722
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• 2007

A new full-length cDNA encoding hyoscyamine $6\beta$-hydroxylase (designated as aah6h, GenBank Accession No. EF187826), which catalyzes the last committed step in the scopolamine biosynthetic pathway, was isolated from young roots of Anisodus acutangulus by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of aah6h was 1380 bp and contained a 1035 bp open reading frame (ORF) encoding a deduced protein of 344 amino acid residues. The deduced protein had an isoelectric point (pI) of 5.09 and a calculated molecular mass of about 38.7 kDa. Sequence analyses showed that AaH6H had high homology with other H6Hs isolated from some scopolamine-producing plants such as Hyoscyamus niger, Datura metel and Atropa belladonna etc. Bioinformatics analyses results indicated AaH6H belongs to 2-oxoglutarate-dependent dioxygenase superfamily. Phylogenetic tree analysis showed that AaH6H had closest relationship with H6H from A. tanguticus. Southern hybridization analysis of the genomic DNA revealed that aah6h belonged to a multi-copy gene family. Tissue expression pattern analysis firstly founded that aah6h expressed in all the tested tissues including roots, stems and leaves and indicated that aah6h was a constitutive-expression gene, which was the first reported tissue-independent h6h gene compared to other known h6h genes.

• 대한수의학회지
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• 제38권1호
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• pp.43-52
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• 1998

서방형 돼지성장호르몬(sustained-release formulation of porcine somatotropin, PST-SR)을 1주 간격으로 6차례 피하 및 근육주사하고 혈액과 조직중의 돼지 성장호르몬(PST)과 insulin-like growth factor 1(IGF-1)의 농도를 측정하여 다음과 같은 결과를 얻었다. 대조군의 혈중 PST와 IGF-1의 농도는 각각 2.41과 95.2 ng/ml 이었다. 1. PST-SR을 투여한 후 PST의 혈중농도는 8시간만에 최대에 도달하여(30 ng/ml) 곧 감소하였다. 혈중농도 반감기(decay half life)는 91~227시간이었다. IGF-1의 혈중농도는 투여후 12시간에 최대에 도달하였으며(165 ng/ml), 이후 서서히 감소되었고 반감기는 77~99시간이었다. 2. 혈중 PST농도-시간의 자료는 제재에서 PST가 유리되는 과정에는 두단계 즉, 투여후 24시간까지의 유리속도가 빠른 단계와 그 이후의 유리속도가 느린 단계가 있음을 보여주었다. 3. 여섯번의 반복투여기간에는 PST의 혈중농도는 투여직후 증가하여 24시간 이후 다음 투여전까지 지속적으로 감소되는 패턴이 반복되었고, 최종투여후 1주일경에는 정상수준으로 회복되었다. 반면에 투여가 반복됨에 따라 매 투여직후의 PST의 혈중 최고치는 다소 증가되는 경향을 보였다(20~40 ng/ml). IGF-1의 혈중농도는 투여가 반복됨에 따라 누적적인 증가현상이 뚜렷하였으며, 이후 2주일후 까지도 정상농도보다 높게 유지되었다(200ng/ml). 임상용량 투여군에서 PST 및 IGF-1의 혈중농도는 투여경로에 따른 차이는 나타나지 않았다. 4. 최종(6번째) 투여후 6, 8, 10, 14일에 조사한 간장, 신장, 소장, 근육, 지방 및 주사부위의 조직중의 PST 농도는 6일째에 이미 대조군 수준으로 회복되었다. IGF-1의 경우 최종투여후 6일에는 간장, 신장, 소장, 지방조직에서 정상보다 높은 농도로 잔류하나 이후 14일까지 모두 대조군 수준으로 감소되었다. 5. 이상의 결과는 본 실험에서 사용된 서방형 PST제제는 최소 1주간 유효성이 유지되며, 동시에 PST는 투여 6일째에, IGF-1은 투여후 14일에 정상수준으로 회복됨을 보여주고 있다.

• 농약과학회지
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• 제19권3호
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• pp.174-184
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• 2015

OECD Test guideline 501에 따라, 벼에서의 [$^{14}C$]butachlor의 식물대사 시험을 실시하였다. 시험토양은 국내 논토양의 대표적인 토성인 양토였으며, 사용지침서상의 사용법에 준하여 이앙 6일 후 [$^{14}C$]butachlor를 입제로 조제하여 1.5 kg/ha 수준으로 토양 수면에 처리하였다. 시료는 시험물질 처리 후 85일과 126일에 채취하였다. 85일 시료는 이삭, 벼 잎, 뿌리로 126일 후 시료는 현미, 왕겨, 볏짚, 뿌리로 분리하였다. 모든 식물 부위의 총 방사성 잔류량은 0.01 mg/kg 이상이었다. 그 후, 대사산물과 모화합물를 확인하기 위하여 극성이 다른 유기용매와 물 혼합용액을 이용하여 시료를 추출 후 Radio-HPLC로 분석하였다. 벼 추출액 중 butachlor는 검출되지 않았고, 5개의 잠정적 주요 대사산물(M1, M2, M3, M4 그리고 M5)이 검출되었으며, 대사산물 중 M4는 2,6-diethylanline로 동정되었다. 토양 추출액에서 N-(butoxymethyl)-N-(2,6-diethyl phenyl)acetamide, 2,6-diethylaniline, M2, M3 그리고 M5가 검출되었으며 butachlor의 농도는 낮은 수준 (약 0.03 mg/kg)으로 검출되었다.

• 생명과학회지
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• 제28권4호
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• pp.389-397
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• 2018

당 단백질에 붙어 있는 당사슬 구조는 형질전환 돼지 유즙으로 분비되는 의약용 단백질의 생물학적 활성, 안정성 그리고 안전성에 영향을 줄 수 있다. 형질전환 동물을 이용한 치료용 당 단백질 생산은 유선 세포에서 이루어지는 당사슬 부가능력에 의해 제한되며, 균일한 당사슬 형태를 가지는 당 단백질 생산은 도전 과제로 남아있다. ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) 유전자는 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 단백질 당화기작에 중요한 효소이지만, 돼지 당 전이효소에 대한 정보는 매우 제한적이다. 따라서, 돼지 B3GNT1 (pB3GNT1) 유전자를 클로닝하고 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 기능적 특성을 조사하였다. 몇가지 다른 프라이머를 사용하여 전체 전사영역(ORF)을 함유하는 부분적인 pB3GNT1 mRNA 염기서열을 간 조직으로부터 분리하였다. 클로닝 된 pB3GNT1의 ORF는 1,248개의 뉴클레오티드를 가지며, 415개 아미노산 잔기로 구성되어 있었다. pB3GNT1 유전자의 장기별 발현특성은 성돈 및 자돈의 여러 기관에서 분석하였다. pB3GNT1 mRNA 발현 수준은 심장, 소장 보다는 근육에서 높았지만 폐에서는 낮았다. pB3GNT1의 기능적 특성 분석을 위해 돼지 신장 세포주(PK-15)에서 pB3GNT1 유전자의 안정적인 발현을 확립하였다. 그 결과, PK-15 세포에서 pB3GNT1 발현에 의한 당화 패턴은 총 시알산 증가에는 영향을 미치지 않지만, poly-N-아세틸글루코사민은 증가하는 것으로 나타났다. 본 연구는 생물반응기로 형질전환 돼지를 이용할 때 희망하는 당사슬을 부가하여 치료 가능성을 높이며 개선된 활성을 나타내는 당단백질 생산에 도움이 될 것이다.

• 한국식품위생안전성학회지
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• 제33권1호
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• pp.58-64
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• 2018

본 연구는 소의 가식부위(근육, 신장, 간장, 지방) 중에서 세팔렉신을 효과적으로 정량분석하기 위한 LC-MS/MS법을 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, $r^2$ > 0.999 이상의 직선성을 나타내었으며, 세팔렉신에 대한 검출한계와 정량한계는 각각 2~10과 $6{\sim}30{\mu}g/kg$으로 나타났다. 또한, 회수율은 83.9~106.8%로 나타났으며, 상대표준편차는 2.3~14.8%로 나타나 정확성이 우수하였다. 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 소의 가식부위 중 세팔렉신을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.

• BMB Reports
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• 제39권3호
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• pp.297-303
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• 2006

As lipolytic enzymes, GDSL lipases play an important role in plant growth and development. In order to identify their functions and roles, the full-length cDNA of a GDSL lipase gene, designated BnLIP2, was isolated from Brassica napus L. BnLIP2 was 1,300 bp long, with 1,122 bp open reading frame (ORF) encoding 373 amino acid residues. Sequence analysis indicated that BnLIP2 belonged to GDSL family. Southern blot analysis indicated that BnLIP2 belonged to a small gene family in rapeseed genome. RT-PCR analysis revealed that BnLIP2 was a tissue-specific expressing gene during reproductive growth and strongly expressed during seed germination. BnLIP2 expression could not be detected until three days after germination, and it subsequently became stronger. The transcript of this gene was deficient in root of seedlings growing at different stages. When juvenile seedlings were treated by methyl jasmonate (MeJ), salicylic acid (SA) and naphthalene acetic acid (NAA), BnLIP2 expression could not be induced in root. Our study implicates that BnLIP2 probably plays an important role in rapeseed germination, morphogenesis, flowering, but independent of root growth and development.