• Mycobiology
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• 제34권4호
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• pp.209-213
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• 2006

Agrocybe cylindracea, an edible mushroom belonging to Bolbitiaceae, Agaricales, is widely used as invaluable medicinal material in the oriental countries. This study was initiated to find the genes expressed during the fruiting body formation of A. cylindracea. The cDNAs expressed differentially during fruiting body morphogenesis of A. cylindracea were isolated through subtractive hybridization between vegetative mycelia and fruiting bodies. The cDNAs expressed in the fruiting body morphogenesis of A. cylindracea were cloned and twenty genes were identified. Eleven were homologous to genes of known functions, three were homologous to genes in other organism without any function known. Six were completely novel genes specific to A. cylindracea so far examined. Some genes with known functions were a pleurotolysin, a self-assembling poreforming cytolysins; Aa-Pril and Pir2p, specifically induced genes during fruiting initiation of other mushroom, Agrocybe aegerita; an amino acid permease; a cytochrome P450; a MADS-box gene; a peptidylprolyl isomerase; and a serine proteinase. For other clones, no clear function was annotated so far. We believe the first report of the differentially expressed genes in fruiting process of A. cylindracea will be great helps for further research.

• 한국응용곤충학회지
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• 제63권1호
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• pp.33-42
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• 2024

복숭아혹진딧물(Myzus persicae)은 다식성으로 담배, 감자, 고추, 배추, 복숭아 등에 심각한 피해를 입히는 대표적인 농업해충이다. 본 연구에서는 국내 복숭아혹진딧물 야외개체군의 λ-cyhalothrin, imidacloprid 및 flupyradifurone에 대한 약제 저항성 발달 수준과 점 돌연변이(R81T, L1014F, M918L)의 발생 여부를 확인하였다. 또한, qRT-PCR을 통해 사이토크롬 P450 유전자인 CYP6CY3 발현량을 확인하였다. 그 결과, λ-cyhalothrin은 저항성비(Resistance Ratio, RR)가 12개 모든 지역이 > 200으로 높은 저항성을 보였다. Imidacloprid와 flupyradifurone은 YS, UR, HY, 그리고 WJ 개체군에서 > 200의 저항성비로 높은 저항성을 나타냈다. R81T는 12개 집단 중 약 50%, L1014F는 약 33.3%, M918L은 100%에서 발현하였다. 또한 qRT-PCR을 통해 imidacloprid 저항성 개체에서 subunit CYP6CY3의 발현량이 높게 나타난 것을 확인하였다. 이러한 결과는 M918L 점 돌연변이는 λ-cyhalothrin 저항성 진단마커로, R81T와 CYP6CY3의 높은 발현은 imidacloprid 저항성 진단마커로 활용이 가능하다는 것을 시사한다.

• 한국미생물·생명공학회지
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• 제38권4호
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• pp.475-480
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• 2010

Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

• Journal of Ginseng Research
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• 제39권2호
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• pp.105-115
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• 2015

Background: Alcoholic steatosis is the earliest and most common liver disease, and may precede the onset of more severe forms of liver injury. Methods: The effect of Korean Red Ginseng extract (RGE) was tested in two murine models of ethanol (EtOH)-feeding and EtOH-treated hepatocytes. Results: Blood biochemistry analysis demonstrated that RGE treatment improved liver function. Histopathology and measurement of hepatic triglyceride content verified the ability of RGE to inhibit fat accumulation. Consistent with this, RGE administration downregulated hepatic lipogenic gene induction and restored hepatic lipolytic gene repression by EtOH. The role of oxidative stress in the pathogenesis of alcoholic liver diseases is well established. Treatment with RGE attenuated EtOH-induced cytochrome P450 2E1, 4-hydroxynonenal, and nitrotyrosine levels. Alcohol consumption also decreased phosphorylation of adenosine monophosphate-activated protein kinase, which was restored by RGE. Moreover, RGE markedly inhibited fat accumulation in EtOH-treated hepatocytes, which correlated with a decrease in sterol regulatory element-binding protein-1 and a commensurate increase in sirtuin 1 and peroxisome proliferator-activated receptor-a expression. Interestingly, the ginsenosides Rb2 and Rd, but not Rb1, significantly inhibited fat accumulation in hepatocytes. Conclusion: These results demonstrate that RGE and its ginsenoside components inhibit alcoholic steatosis and liver injury by adenosine monophosphate-activated protein kinase/sirtuin 1 activation both in vivo and in vitro, suggesting that RGE may have a potential to treat alcoholic liver disease.

• 동의생리병리학회지
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• 제17권1호
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• pp.91-100
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• 2003

In the present study, we investigated the protective effect of the Lycii Fructus water extracts (LFE) against CCl4-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of LFE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation, In addition, the pretreatment of LFE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The LFE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of LFE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the LFE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

• 동의생리병리학회지
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• 제17권2호
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• pp.297-307
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• 2003

In the present study, we investigated the protective effect of the Puerarie Radix water extract (PRE) against CCl₄-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of PRE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation. In addition, the pretreatment of PRE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The PRE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of PRE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the PRE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

• Environmental Analysis Health and Toxicology
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• 제21권3호
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• pp.291-299
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• 2006

The experiments investigated whether early exposure to testosterone propionate (TP) during prepuberty alters testis development in Sprague-Dawley male rats. We performed Hershberger assay using the stimulated weanling male rats by OECD protocols, cDNA microarray, and Western blot. TP was subcutaneously injected to uncastrated Sprague-Dawley male rat of 22 days old for 10 consecutive days at doses of 0.4, 0.8, 1.0, 1.2, 1.6 mg/kg per day. At necropsy, the following tissues were removed and weighed: combined testes, epididymides (Epi), Cowper's glands (COW), levator am, and bulbocavernosus muscles (LABC), seminal vesicles, together with coagulating gland (SV) and ventral prostate (VP). We found that TP increased the weights of Epi, VP, SV, COW, and LABC, while testis was decreased in a dose-dependent manner. In cDNA microarray analysis of testis, there were significant reductions in the expression of cytochrome P450 11A (CYP11A), the rate-limiting enzyme of steroidogenesis. Taken together these results, TP exposure before puberty in male rats may produce the delay in testis development by inhibiting the CYP11A gene expression.

• Journal of Ginseng Research
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• 제44권1호
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• pp.123-134
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• 2020

Background: The lepidopteran Asiatic corn borer (ACB), Ostrinia furnacalis (Guenee), has caused huge economic losses throughout the Asian-Western Pacific region. Usually, chemical pesticides are used for the control, but excessive use of pesticides has caused great harm. Therefore, the inartificial ecotypic pesticides to ACB are extremely essential. In our previous study, we found that panaxadiol saponins (PDS) can effectively reduce the harm of ACB by causing antifeedant activity. Therefore, it is necessary to reveal the biological molecular changes in ACB and the functionary mechanism of PDS. Methods: We analyzed the global transcription of ACB with different PDS concentration treatment (5 mg/mL, 10 mg/mL, and 25 mg/mL) by high-throughput sequencing and de novo transcriptome assembly method. Results: PDS treatment could cause the changes of many gene expressions which regulate its signal pathways. The genes in peroxisome proliferator-activated receptor (PPAR) signaling pathway were significantly downregulated, and then, the downstream fatty acid degradation pathway had also been greatly affected. Conclusion: Through this experiment, we hypothesized that the occurrence of antifeedant action of ACB is because the PDS brought about the downregulation of FATP and FABP, the key regulators in the PPAR, and the downregulation of FATP and FABP exerts further effects on the expression of SCD-1, ACBP, LPL, SCP-X, and ACO, which leads to the disorder of PPAR signaling pathway and the fatty acid degradation pathway. Not only that, PDS treatment leads to enzyme activity decrease by inhibiting the expression of genes associated with catalytic activity, such as cytochrome P450 and other similar genes.

• KSBB Journal
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• 제29권1호
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• pp.58-66
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• 2014

Demand for in vitro pharmacological evaluation and toxicity test using human hepatocytes has been increasing. In USA and Europe, human hepatocytes obtained from donated whole liver unsuitable for transplantation were distributed to researchers and deposited in cell bank facility as cryopreserved vial. In Korea, however, incidence of transplantation- inappropriate whole liver has been quite low and the whole livers almost have so severe liver disease such as fatty or fibrotic liver that cannot meet the demand. In this study we aimed to isolate human hepatocytes from liver resection surgery-originated partial liver, and assure the isolated human hepatocytes and its cryopreserved hepatocytes to be qualified for the in vitro pharmacological evaluation and drug toxicity tests. We compared those with commercially available human hepatocyte, BD $GenTest^{TM}$ by cell morphology, hepatic gene expression, urea synthesis, albumin secretion, ammonia removal, and cytochrome P450 induction activities. Changes in hepatotoxic gene expression after cryopreservation are evaluated with a typical hepatotoxic drug, acetaminophen. Consequently, the fresh hepatocytes from the partial liver and its cryopreserved hepatocytes expressed their intrinsic hepatic functions well and showed equal hepatotoxicity gene expression trend regardless to cryopreservation. Therefore, liver resection surgery-originated partial liver can be used as a useful source of human hepatocytes for various pharmacological and hepatotoxicity test.

• 한국작물학회지
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• 제67권1호
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• pp.41-52
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• 2022

1. 본 연구에서는 우리밀 품종인 금강밀과 염 저항성을 가지는 돌연변이 라인 2020-s1340을 재료로 200 mM 염 스트레스 처리에 따른 전사체 발현을 확인하였다. QuantSeq을 통해 23,634,438개의 reads가 생산되었고 7,331,269개의 reads가 mapping됐다. 2. 염 스트레스 상황에서 총 282개의 DEG가 확인이 되었고 이러한 DEGs는 UDP-glucosyltransferase, receptor kinase-like protein, Lectin receptor-like kinases, cytochrome P450등의 단백질들을 코딩하는 유전자들이다. 이러한 DEGs는 염 저항성과 관련된 후보 유전자들이 될 수 있다. 염 저항성과 관련하여 역할이 밝혀지지 않은 유전자들은 추후 연구를 통해 확인이 필요하다. 3. GO연구에서는 DEGs를 세가지 범주로 분류하였으며 대부분 식물체 내 세포 기초 경로와 관련된 GO term들이 주로 되었으며 각각 범주에 있어서 biological process, molecular process에서는 single-organism process (GO: 0044699), single-organism metabolic process (GO:0044710), oxidation-reduction process (GO:0055114), copper ion transport (GO:0006825), copper ion transmembrane transport (GO:0035434), alternative oxidase activity (GO:0009916) GO term들이 유의성이 높게 나타났다. 4. 이러한 QuantSeq의 분석결과는 밀에 관한 염에 의해 발현되는 전사 발현에 대한 이해를 향상시킬 수 있다. 또한 염 스트레스 반응의 복잡한 분자 메커니즘에 대한 좋은 통찰력을 제공하고 염분 스트레스에 대한 작물 내성의 유전적 개선을 위한 실질적인 토대를 마련할 수 있을 것이다.