• Journal of Pharmaceutical Investigation
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• 제26권1호
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• pp.33-41
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• 1996

In order to study the effect of phenobarbital(PB) on the hepatic transport of diltiazem(DTZ), $Ca^{2+}$ channel blocker, we used isolated hepatocytes of rat which was intraperitoneally pretreated with phenobarbital sodium(75 mg/kg) for four days once a day. For the isolation of rat liver cells, a modification of the two step procedure of Seglen was used. DTZ was dissolved in incubation buffer to the final DTZ concentrations of 200, 400, 600, 800 and 1000 ng/ml in order to elucidate the uptake characteristics of DTZ by hepatocytes. Reactions were stopped at 10, 20, 30, 45, 60, 90, 120 and 300 sec. The initial velocity was determined by disappearance of diltiazem in the hepatocyte suspension. On the other hand, to determine the effect of PB on the in vitro hepatic intrinsic clearance of DTZ we obtained the metabolism rates of DTZ in the control and the PB-pretreated rat hepatocyte at various time intervals. According to pretreatment with PB, the size of hepatocyte and the amount of protein per $10^6$ cells were significantly (p<0.01) increased from $26.92{\pm}0.1364\;m$ to $35.31{\pm}1.00\;m$ and from $468{\pm}6.5\;{\mu}g/10^6$ cells to $628.8{\pm}12.1{\mu}g/10^6$ cells, respectively. In the case or hepatic uptake of diltiazem, $K_m$ was not different in the normalization by cell numbers and increased from $2.90\;{\mu}M\;to\;13.89\;{\mu}M$ in the normalization by protein amount. $V_max$ was increased regardless of normalization by protein amount and cell numbers, from $1.21\;{\mu}mole/min \;{\cdot}\;mg\;protein\;to\;3.96\;{\mu}mole/min\;{\cdot}\;mg\;protein\;and\;from\;2.38\;{\mu}mole/min\;{\cdot}\;10^6\;cells\;to\;2.83\;{\mu}mole/min\;{\cdot}\;10^6\;cells$, respectively. The in vitro hepatic intrinsic clearance of DTZ was significantly (p<0.01) increased from $0.640{\pm}0.038\;ml/mim\;{\cdot}\;10^6\;cells\;to\;2.385{\pm}0.212\;ml/min\;{\cdot}\;10^6\;cells$ due to PB-pretreatment. These results suggest that the uptake of DTZ by hepatocyte is extremely fast and PB enhances the hepatic intrinsic metabolic clearance of DTZ.

• Journal of Ginseng Research
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• 제35권2호
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• pp.209-218
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• 2011

Ginseng has been used as a general tonic agent to invigorate the human body as an adaptogenic agent. In a previous report, we have shown that ginseng contains a novel glycolipoprotein called gintonin. The main function of gintonin is to transiently enhance intracellular free $Ca^{2+}$ $[Ca^{2+}]_i$ levels in animal cells. The previous method for gintonin isolation included multiple steps using organic solvents. In the present report, we developed a simple method for the preparation of crude gintonin from ginseng root as well as stem and leaf, which produced a higher yield of gintonin than the previous one. The yield of gintonin was 0.20%, 0.29%, and 0.81% from ginseng root, stem, and leaf, respectively. The apparent molecular weight of gintonin isolated from stem and leaf through sodium dodecyl sulfate polyacrylamide gel electrophoresis was almost same as that from root but the compositions of amino acids, carbohydrates or lipids differed slightly between them. We also examined the effects of crude gintonin from ginseng root, stem, and leaf on endogenous $Ca^{2+}$-activated $Cl^-$ channel (CaCC) activity of Xenopus oocytes through mobilization of $[Ca^{2+}]_i$. We found that the order of potency for the activation of CaCC was ginseng root > stem > leaf. The $ED_{50}$ was $1.4{\pm}1.4$, $4.5{\pm}5.9$, and $3.9{\pm}1.1$ mg/mL for root, stem and leaf, respectively. In the present study, we demonstrated for the first time that in addition to ginseng root, ginseng stem and leaf also contain gintonin. Gintonin can be prepared from a simple method with higher yield of gintonin from ginseng root, stem, and leaf. Finally, these results demonstrate the possibility that ginseng stem and leaf could also be utilized for ginstonin preparation after a simple procedure, rather than being discarded.

• Asian-Australasian Journal of Animal Sciences
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• 제32권4호
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• pp.494-500
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• 2019

Objective: Feed consumption contributes a large percentage for total production costs in the poultry industry. Detecting genes associated with feeding traits will be of benefit to improve our understanding of the molecular determinants for feed efficiency. The objective of this study was to identify candidate genes associated with feed conversion ratio (FCR) via genomewide association study (GWAS) using sequence data imputed from single nucleotide polymorphism (SNP) panel in a Chinese indigenous chicken population. Methods: A total of 435 Chinese indigenous chickens were phenotyped for FCR and were genotyped using a 600K SNP genotyping array. Twenty-four birds were selected for sequencing, and the 600K SNP panel data were imputed to whole sequence data with the 24 birds as the reference. The GWAS were performed with GEMMA software. Results: After quality control, 8,626,020 SNPs were used for sequence based GWAS, in which ten significant genomic regions were detected to be associated with FCR. Ten candidate genes, ubiquitin specific peptidase 44, leukotriene A4 hydrolase, ETS transcription factor, R-spondin 2, inhibitor of apoptosis protein 3, sosondowah ankyrin repeat domain family member D, calmodulin regulated spectrin associated protein family member 2, zinc finger and BTB domain containing 41, potassium sodium-activated channel subfamily T member 2, and member of RAS oncogene family were annotated. Several of them were within or near the reported FCR quantitative trait loci, and others were newly reported. Conclusion: Results from this study provide valuable prior information on chicken genomic breeding programs, and potentially improve our understanding of the molecular mechanism for feeding traits.

• Fisheries and Aquatic Sciences
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• 제24권11호
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• pp.360-369
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• 2021

Paralytic shellfish toxins (PSTs) and tetrodotoxin (TTX) are neurotoxins that display pharmacological activity that is similar to that of specific sodium channel blockers; they are the principle toxins involved in shellfish and puffer fish poisoning. In Korea, puffer fish is a very popular seafood, and several cases of accidental poisoning by TTX have been reported. Therefore, it is necessary to determine whether puffer fish poisoning incidents are caused by PSTs or by TTX. In this study, we used mouse bioassay (MBA) and liquid chromatograph-tandem mass spectrometry (LC-MS/MS) to determine the presence of PSTs and TTX in puffer fish from an area near Mireuk-do, Tong-Yeong on the southern coast of Korea from January through March, 2014. The toxicity of PSTs and TTX extracts prepared from three organs of each specimen was analyzed by MBA. Most of the extracts killed mice with typical signs of TTX and PSTs. The LC-MS/MS analysis of seven specimens of Takifugu pardalis and Takifugu niphobles, each divided into muscles, intestines, and liver, were examined for TTX. In T. pardalis, the TTX levels were within the range of 1.3-1.6 ㎍/g in the muscles, 18.8-49.8 ㎍/g in the intestines, and 23.3-96.8 ㎍/g in the liver. In T. niphobles, the TTX levels were within the range of 2.0-4.5 ㎍/g in the muscles, 23.9-71.5 ㎍/g in the intestines, and 28.1-114.8 ㎍/g in the liver. Additionally, the toxicity profile of the detected PSTs revealed that dcGTX3 was the major component in T. pardalis and T. niphobles. When PSTs were calculated as saxitoxin equivalents the levels were all less than 0.5 ㎍/g, which is below the permitted maximum standard of 0.8 ㎍/g. These findings indicate that the toxicity of T. pardalis and T. niphobles from the southern coast of Korea is due mainly to TTX and that PSTs do not exert an effect.

• 한국응용곤충학회지
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• 제53권2호
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• pp.149-156
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• 2014

2009~2011년 동안 국내 주요 배추 재배지 5개 지역(평창, 홍천, 봉화, 무주, 제주)에서 살충제에 대한 복숭아혹진딧물의 저항성 발달 정도를 조사하고, 야외 개체군에 적용 가능한 살충제 저항성 마커를 개발하기 위해 본 연구를 수행하였다. 조사된 5개 지역 개체군 모두 여러 살충제 종류에 대하여 다양한 저항성을 보였다. 다양한 저항성을 보인 5개 지역 야외 개체군으로부터 여라 살충제에 대하여 복합적으로 저항성을 보이는 복합저항성 계통(MR)을 선발하였고, 이 MR과 모든 지역 채집 개체군에 대해 등전점전기영동과 정량염기서열분석(quantitative sequencing, QS)을 통하여 에스터레이즈 과발현과 살충제 작용점 내 돌연변이를 확인하였다. MR을 포함한 모든 야외 개체군에서 에스터레이즈의 과발현과 아세틸콜린에스터레이즈 1 유전자(ace1)의 StoF 돌연변이를 확인할 수 있었다. 넉다운 저항성 돌연변이로 잘 알려진 파라 타입 나트륨 채널 유전자(para)의 LtoF 돌연변이는 모든 지역 채집 개체군은 물론 비펜스린에 대해 3,461배 저항성을 보이는 MR에서도 발견되지 않았다. 그 외에 MtoL 돌연변이를 발견하였는데, 생물검정 결과 저항성 수준과 돌연변이 발생 빈도가 일치하였다. 따라서 생물검정 대신, 이러한 분자 마커를 활용 한다면 더 효율적으로 살충제 저항성 평가가 가능할 것이다. 이러한 분자 마커들(ace1의 StoF, para의 MtoL)은 정량염기서열분석, PCR amplification of specific alleles (PASA) 등의 진단 방법에 쉽게 응용이 가능 하고, 이러한 방법은 야외 복숭아혹진딧물 저항성 관리에 적용이 가능 할 것이다.

• Journal of Chest Surgery
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• 제41권4호
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• pp.405-416
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• 2008

배경: 흉부 및 흉복부 대동맥의 수술 중 대동맥 차단은 허혈성 척수 손상에 의한 하반신 마비와 같은 심각한 합병증을 유발할 수도 있어 수술 중 허혈성 척수손상을 예방하기 위한 여러 방법의 연구가 계속 되고 있다. 최근에 허혈성 대뇌 손상 모델에서 신경조직의 막전위 의존성 나트륨채널 길항제가 대뇌 보호 효과가 있다는 보고가 있다. 본 연구는 토끼의 허혈성 척수손상 모델에서 나트륨채널 길항제인 페니토인과 저체온의 척수보호효과를 비교해 보고자 시행되었다. 대상 및 방법: 뉴질랜드산 토끼의 신동맥직하부에서 복부대동맥을 25분간 차단하는 방식으로 척수허혈을 유도하였으며 각 군당 8마리씩 네 군으로 나누었다. 대조군과(S39) 저체온군은(S37) 대동맥 차단시간 동안 직장온도를 각기 $39^{\circ}C$와 $37^{\circ}C$로 일정하게 유지하면서 $22^{\circ}C$ 생리적 식염수만 2 mL/min의 속도로 연속 주입하였으며, 정상체온 및 저체온 페니토인 군은(P39, P37) 앞의 두 군과 동일한 방법으로 하되 생리적 식염수에 페니토인을 녹여 주입하였다(100 mg/50 mL). 수술 후 24시간 및 72시간이 경과한 다음 Tarlov scoring을 통해 신경학적 평가를 시행하였고 마지막 평가 후에는 객관적으로 신경손상의 정도를 정량화하기 위해 척수를 고정 처리하였다. 결과: 페니토인의 역행성 주입에 따른 심각한 문제는 없었으며 대조군에(S39) 속한 모든 동물은 완전 또는 심한 하반신 마비 소견을 보였다. 페니토인과(P39) 저체온(S37)군 모두 대조군에 비해 신경학적 평가는 유사한 정도로 우수한 결과를 보였다(p<0.05). 조직 병리학적 검사 결과, 대조군에 속한 모든 동물은 척수 회백질에서 심한 신경조직 괴사 때 보이는 전형적인 특징을 보여주었으며, TUNEL 염색에 양성인 신경세포도 높은 빈도로 관찰되었으나, 저체온 또는 페니토인 투여 군에서는 괴사현상이 유의한 정도로 감소하였으며, 상대적으로 매우 낮은 빈도의 TUNL 염색 양성세포가 관찰되었다(p<0.05). 그러나 저체온과 페니토인을 병용했을 때의 부가적인 척수보호효과를 조사해 본 결과 신경학적 평가와 조직병리학적 결과 모두 유의한 수준의 부가적인 효과는 없었다. 걸론: 결론적으로, 토끼의 허혈성 척수 손상 모델을 이용하여 페니토인과 저체온의 신경보호효과를 알아본 결과 신경학적 평가와 조직병리학적 검사 결과 모두 부가적인 효과는 보여주지 못했지만 각각의 경우 유사한 정도의 신경보호효과를 보여주었다.