• 제목/요약/키워드: NADPH-generating system

검색결과 17건 처리시간 0.032초

Convenient Assay of O2- Generated on Potato Tuber Tissue Slices Treated with Fungal Elicitor by Electron Spin Resonance - No Secondary Oxidative Burst Induction by H2O2 Treatment

  • Park, Hae-Jun;Doke, Noriyuki
    • The Plant Pathology Journal
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    • 제21권3호
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    • pp.283-287
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    • 2005
  • Since the discovery of generation of $O_2^-$ in plant, many evidence for the oxidative burst (OXB) has been accumulated in various combinations of plant and pathogen or elicitor systems. $O_2^-$ generating system responsible for the OXB was coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in microsomal fraction isolated from sliced aged potato tuber slices which were treated by hyphal wall components elicitor from Phytophthora infestans (HWC). We developed new assay method for quantitative measurement of oxygen radical $O_2^-$ by using electron spin resonance (ESR) analysis during elicitor­induced OXB on the surface of plant tissues. The ESR analysis using an $O_2^-$ trapper, Tiron (1,2-dihydroxy-3,5­benzenedisulfonic acid), provided a convenient assay for detecting only $O_2^-$ during elicitor-induced OXB producing various active oxygen species (AOS) on plant tissue surface. Tiron was oxidized to Tiron semiquinon radical by $O_2^-$. Quantity of the radical signal was measured by specific spectra on ESR spectroscopy. The level of $O_2^-$ was high in from surface of potato tuber tissue treated with hyphal cell wall elicitor (HWC) from Phytophthora infestans. There was no secondary OXB induction by $H_2O_2$ treatment in plant.

돼지 간중의 Monooxygenase 가 Diazinon 의 분해에 미치는 영향 (The degradation o Diazinon by hepatic monooxygenase of Pig)

  • 류종국;이규승
    • 한국환경농학회지
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    • 제9권2호
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    • pp.153-159
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    • 1990
  • 돼지간을 두 fraction($105,000{\time}g$ pellet, superatant)으로 나누어 Diazinon의 분해에 관여하는 두 효소(Monooxygenase와 esterase)의 활성 및 저해제(EPN, Beam, PBO)첨가에 따른 Diazinon의 분해억제 능력을 비교 검토하였다. 1. 돼지간의 microsomal fraction과 soluble fraction에서 M. O.와 β-esterase 활성은 20분과 60분에 최고활성을 나타내었다. 2. Microsomal fraction에서의 M. O.활성은 NADPH generating system의 첨가로 1.6배의 활성증가와 지속성이 확인되었다. 3. Microsomal fraction에서의 Diazinon분해량은 30분에 최고량(0.060nM/ mg/ min)을 보였으나, soluble fraction에서는 60분에 최고량(0.018nM/ mg/ min)을 보여 1/3수준이었다. 4. M. O.활성에 미치는 Diazinon에 대한 EPN, Beam, PBO의 첨가효과는 Beam(14%)< EPN(15%)

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Comparative GC-MS Based In vitro Assays of 5α-Reductase Activity Using Rat Liver S9 Fraction

  • Lee, Su-Hyeon;Lee, Dong-Hyoung;Lee, Jeong-Ae;Lee, Won-Yong;Chung, Bong-Chul;Choi, Man-Ho
    • Mass Spectrometry Letters
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    • 제3권1호
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    • pp.21-24
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    • 2012
  • $5{\alpha}$-Dihydrotestosterone (DHT) is the primary active metabolite of testosterone, catalyzed by $5{\alpha}$-reductase ($5{\alpha}R$) in the skin, prostate, and liver. In this study, the $5{\alpha}R$ activity in rat liver S9 fraction in the presence of a NADPH-generating system was evaluated and compared by gas chromatography-mass spectrometry (GC-MS)-based in vitro assays. Testosterone and a $5{\alpha}R$ inhibitor, finasteride, were added to the S9 fractions and incubated at $37^{\circ}C$ for 1 h. Both testosterone and DHT were quantitatively measured and compared with two different GC-MS-based steroid profiling techniques. DHT was not detected by conventional GC-MS analysis in the absence of finasteride when the concentration of testosterone in the S9 fraction was less than $0.2{\mu}M$, whereas the isotope-dilution GC-MS (GC-IDMS) system was able to evaluate the $5{\alpha}R$ activity. Because the S9 fraction contains more reactive enzymes and is easier to collect from tissues compared with a microsomal solution, the combination of the S9 fraction and GC-IDMS technique may be a promising assay for evaluating the $5{\alpha}R$ activity in large-scale clinical studies.

잡초종(雜草種)에 대한 Paraquat 독성(毒性) : 내성종(耐性種)과 감수성(感受性) 종간(種間)의 반응(反應) 차이(差異) (Paraquat Toxicity in Weed Species : Difference in Physiological Responses between Tolerant and Susceptible Species)

  • 강병화;심상인
    • 한국잡초학회지
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    • 제15권3호
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    • pp.224-231
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    • 1995
  • Paraquat에 대한 잡초종(雜草種)의 다양한 반응(反應)은 여러 가지 요인에 의한다. Paraquat에 대하여 내성종(耐性種)과 감수성종(感受性種)은 가시적(可視的)인 피해율(被害率)의 차이(差異)를 비롯하여 체내(體內) 물질(物質) 변화(變化)의 차이(差異)는 물론 체내(體內) 효소(酵素) 활성(活性)의 차이(差異) 등을 나타났다. 내성종(耐性種)은 paraquat에 의한 photosystem II의 활성(活性) 저하(低下) 정도가 감수성종(感受性種)보다 작았으며, 지질(脂質)의 과산화(過酸化) 정도(程度) 역시 내성종(耐性種)에서 낮았다. 체내(體內) 물질(物質) 중 엽록소(葉綠素)는 paraquat에 의해 줄어들었으나 환원당(還元糖)의 경우는 paraquat의 농도(濃度)가 높아질수록 증가하였다. 항산화제(抗酸化劑)인 glutathione의 경우 처리농도(處理濃度)가 증가함에 따라 함량(含量)이 증가하는 경향이었으나 내성종(耐性種)과 감수성종(感受性種)의 구분은 뚜렷하지 않았다. 항산화효소(抗酸化酵素)인 glutathione reductase의 활성(活性)은 내성종(耐性種)에서 높게 나타났으며, superoxide dismutase의 경우는 paraquat의 처리농도(處理濃度)가 증가함에 따라 그 활성(活性)이 증가하였다. 체내대사(體內代謝)의 주요 효소(酵素)들인 catalase, NADPH-cytochrome c reductase, malate dehydrogenase의 활성(活性)은 catalase의 경우 저농도(低濃度)의 paraquat에 의해서는 활성(活性) 저하(低下) 정도(程度)가 낮은 반면 나머지 두 효소(酵素)는 저농도(低濃度)에서부터 급격한 활성저하(活性低下)가 나타났다.

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황백의 주요 구성 화합물에 의한 약물대사효소 및 약물수송단백 저해능 평가 (Inhibition of Drug-metabolizing Enzyme and Drug Transporter by Major Components of Phellodendri cortex)

  • 구혜영;김현미;손지홍;유광현
    • 한국해양바이오학회지
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    • 제1권3호
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    • pp.213-217
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    • 2006
  • 본 연구는 황백에 함유되어 있는 주요 화합물인 berberine, palmatine, limonin 및 rutaecarpine의 CYP2D6 및 p-glycoprotein 활성에 대한 저해정도를 탐색함으로써, 황백을 다른 양약과 병용시 약물상호작용을 유발할 수 있는 가능성을 평가하고자 하였다. 인체 간 마이크로좀 시료에 CYP2D6 동효소의 기질약물인 dextromethorphan과 NADPH 재생성계 및 저해제 ($200{\mu}M$)를 첨가한 후 반응시켜 생성된 대사물을 LC/MS/MS를 이용하여 정량하여 CYP2D6 동효소 활성의 변화를 평가하였다. 또한 약물수송단백인 p-glycoprotein의 활성은 L-MDR1 세포주를 이용한 calcein AM 축적 실험을 통하여 평가하였다. 그 결과 식물 알카로이드인 berberine에서 강력한 CYP2D6 활성 저해능을 관찰하였으며, 저해 효과는 농도 의존적으로 증가하였으며, mechanism-based 저해 기전을 나타내었다. 그러나 limonine과 rutaecarpine은 CYP2D6 저해 활성을 보이지 않았고, p-glycoprotein 기능에 대해서는 평가한 어떤 화합물도 저해 활성을 나타내지 않았다. 황백의 주요 성분인 berberine의 CYP2D6 활성 저해능을 고려할 때, 황백을 CYP2D6 기질약제와 병용시 약물상호작용을 유발할 가능성을 보여준다. 이러한 황백의 CYP2D6를 매개로한 임상적인 약물상호작용 가능성은 임상시험을 통하여 추가적인 검정이 필요할 것으로 사료된다.

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A case report of chronic granulomatous disease presenting with aspergillus pneumonia in a 2-month old girl

  • Lee, Eun;Oh, Seak-Hee;Kwon, Ji-Won;Kim, Byoung-Ju;Yu, Jin-Ho;Park, Chan-Jeoung;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • 제53권6호
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    • pp.722-726
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    • 2010
  • Chronic granulomatous disease (CGD) is an uncommon inherited disorder caused by mutations in any of the genes encoding subunits of the superoxide-generating phagocyte NADPH oxidase system, which is essential for killing catalase producing bacteria and fungi, such as $Aspergillus$ species, $Staphylococcus$ $aureus$, $Serratia$ $marcescens$, $Nocardia$ species and $Burkholderia$ $cepacia$. In case of a history of recurrent or persistent infections, immune deficiency should be investigated. Particularly, in the case of uncommon infections such as aspergillosis in early life, CGD should be considered. We describe here a case of CGD that presented with invasive pulmonary aspergillosis in a 2-month-old girl. We confirmed pulmonary aspergillosis noninvasively through a positive result from the culture of bronchial alveolar lavage fluid, positive serological test for $Aspergillus$ antigen and radiology results. She was successfully treated with Amphotericin B and recombinant IFN-${\gamma}$ initially. Six weeks later after discharge, she was readmitted for pneumonia. Since there were infiltrates on the right lower lung, which were considered as residual lesions, voriconazole therapy was initiated. She showed a favorable response to the treatment and follow-up CT showed regression of the pulmonary infiltrates.

Pharmacokinetics of LB20304, a New Fluoroquinolone, in Rats and Dogs

  • Seo, Mi-Kyeong;Lee, Sun-Hwa;Choi, Yun-Jeong;Jeong, Yi-Na;Lee, Sung-Hack;Kim, In-Chull;Lee, Yong-Hee
    • Archives of Pharmacal Research
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    • 제19권5호
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    • pp.359-367
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    • 1996
  • The pharmacokinetics of LB20304 was investigated following intravenous (IV) and oral administration to rats and dogs. Additionally, in vitro metabolism and serum protein binding studies were also conducted. The total body clearance, apparent volume of distribution, terminal half-life, and extent of bioavailability were 21.8 ml/min/kg, 2265 ml/kg, 93.6 min, and 30.8% for rats; and 7.95 ml/min/kg, 4144 ml/kg, 363 min, and 81.1% for dogs, respectively. LB20304 was stable in the liver microsome containing NADPH generating system and its serum protein binding was 58.5-65.8% for rats, 19.1-29.6% for dogs, and 56.9-59.6% for humans. Its tissue concentration levels in lever, stomach, small intestine, and kidney were 9.5 to 26.1 times greater than plasma level, but the concentration in testis was quite low and that in brain was negligible in rats. The 48 hr urinary recovery of the dose was 44% for IV dosing and 14% for oral dosing, shereas the 48 hr biliary recovery of the dose was 6.4% for IV dosing and 4.5% for oral dosing in rats. In summary, the pharmacokinetic properties of LB20304 were characterized by its good oral absorption, long plasma half-life, and good tissue distribution.

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