• Title/Summary/Keyword: nad5 gene

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Complete Mitochondrial Genome of Haplorchis taichui and Comparative Analysis with Other Trematodes

  • Lee, Dongmin;Choe, Seongjun;Park, Hansol;Jeon, Hyeong-Kyu;Chai, Jong-Yil;Sohn, Woon-Mok;Yong, Tai-Soon;Min, Duk-Young;Rim, Han-Jong;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.719-726
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    • 2013
  • Mitochondrial genomes have been extensively studied for phylogenetic purposes and to investigate intra- and interspecific genetic variations. In recent years, numerous groups have undertaken sequencing of platyhelminth mitochondrial genomes. Haplorchis taichui (family Heterophyidae) is a trematode that infects humans and animals mainly in Asia, including the Mekong River basin. We sequenced and determined the organization of the complete mitochondrial genome of H. taichui. The mitochondrial genome is 15,130 bp long, containing 12 protein-coding genes, 2 ribosomal RNAs (rRNAs, a small and a large subunit), and 22 transfer RNAs (tRNAs). Like other trematodes, it does not encode the atp8 gene. All genes are transcribed from the same strand. The ATG initiation codon is used for 9 protein-coding genes, and GTG for the remaining 3 (nad1, nad4, and nad5). The mitochondrial genome of H. taichui has a single long non-coding region between trnE and trnG. H. taichui has evolved as being more closely related to Opisthorchiidae than other trematode groups with maximal support in the phylogenetic analysis. Our results could provide a resource for the comparative mitochondrial genome analysis of trematodes, and may yield genetic markers for molecular epidemiological investigations into intestinal flukes.

Role of Osmotic and Salt Stress in the Expression of Erythrose Reductase in Candida magnoliae

  • Park, Eun-Hee;Lee, Ha-Yeon;Ryu, Yeon-Woo;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1064-1068
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    • 2011
  • The osmotolerant yeast, Candida magnoliae, which was isolated from honeycomb, produces erythritol from sugars such as fructose, glucose, and sucrose. Erythrose reductase in C. magnoliae (CmER) reduces erythrose to erythritol with concomitant oxidation of NAD(P)H. Sequence analysis of the 5'-flanking region of the CmER gene indicated that one putative stress response element (STRE, 5'-AGGGG-3'), found in Saccharomyces cerevisiae, exists 72 nucleotides upstream of the translation initiation codon. An enzyme activity assay and semiquantitative reverse transcription polymerase chain reaction revealed that the expression of CmER is upregulated under osmotic and salt stress conditions caused by a high concentration of sugar, KCl, and NaCl. However, CmER was not affected by osmotic and oxidative stress induced by sorbitol and $H_2O_2$, respectively. The basal transcript level of CmER in the presence of sucrose was higher than that in cells treated with fructose and glucose, indicating that the response of CmER to sugar stress is different from that of GRE3 in S. cerevisiae, which expresses aldose reductase in a sugarindependent manner. It was concluded that regulation of CmER differs from that of other aldose reductases in S. cerevisiae.

Induction of the Proteasome Subunits by Xanthohumol Compounds from Hops (Hop 유래 Xanthohumol 화합물에 의한 Proteasome계의 유도발현)

  • Lee, Hyang-Rim;Lee, Yong-Rok;Kwak, Mi-Kyoung
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.481-487
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    • 2010
  • The proteasome plays a major role in the degradation of abnormal proteins within the cell. Therefore, repressed proteasome function is accepted as one of factors contributing the pathogenesis of multiple degenerative diseases. In the present study, we have observed that xanthohumol C, which is one of prenylated flavonoids from hops, increases the expression of the proteasome subunits through the Nrf2 pathway. Treatment of murine renal epithelial TCMK-1 cells with xanthohumol C and its methoxymethoxy-derivative elevated the expression of the Antioxidant Response Element (ARE)-driven reporter gene, as well as Nrf2-target genes including NAD(P)H: quinoneoxidoreductaes 1 (Nqo1). Transcript levels for the catalytic subunits of the proteasome Psmb5 and Psmb6 were increased by these compounds. The activation of the psmb5 promoter by xanthohumol C was abolished when the ARE in this promoter was mutated, indicating that proteasome induction was mediated by the Nrf2-ARE pathway. These results suggest that xanthohumol compounds from hops have a potential benefit on various oxidative stress-associated human diseases through the induction of the proteasome.

Two Human Cases of Echinococcus ortleppi Infection in the Lung and Heart in Vietnam

  • De, Nguyen Van;Minh, Pham Ngoc;Duyet, Le Van;Bich, Nguyen Ngoc;Son, Trinh Nam;Jung, Bong-Kwang;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.58 no.4
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    • pp.451-456
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    • 2020
  • This is a report of 2 cases of human hydatidosis caused by Echinococcus ortleppi in Vietnam. The patients were a 12-year-old male (case 1) having a cyst of 10.0×9.0 cm size in the lung and a 50-year-old female with a 3.0×3.3 cm-sized cyst in the heart. Eosinophilia was 33.7% in the male and 45.8% in the female patient. C-reactive protein was increased to 16.5 mg/L in the male and 18.2 mg/L in the female. Both patients were positive for ELISA at OD=2.5 and 3.1, respectively. Echinococcus protoscolices were collected from the cysts by amniocentesis and surgery. The protoscolices were identified as E. ortleppi by morphology and analysis of mitochondrial NADH dehydrogenase 1 (nad1) gene sequence. Both patients were cured by surgical resection of the hydatid cyst combined with albendazole medication. The E. ortleppi infection in lung is the second report, and the other in the heart is the first in Vietnam.

Monitoring of the Distribution of Ambient Air Particles in Seoul Using a Cascade Impactor and the Particle Toxicity

  • Park, Eun-Jung;Kim, Dae-Seon;Park, Kwang-Sik
    • Environmental Analysis Health and Toxicology
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    • v.25 no.2
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    • pp.99-109
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    • 2010
  • The distribution of ambient air particles varies according to climate, industries, and other sources. In this study, ambient air particles (less than 12.1 ${\mu}m$) were monitored from February to August, 2007 as 12 different fractions sorted by a cascade impactor. Particles in the size range from 0.33 ${\mu}m$ to 0.76 ${\mu}m$ comprised the main fraction of ambient air particles in Seoul, Korea. On the day of an Asian dust event, the particle fraction size increased to 1.25~2.5 ${\mu}m$. The different sized particle fractions were also monitored for metals and were found to contain toxic heavy metals including Pb, Cd, Hg, Cr and As. Particle preparations were significantly cytotoxic when exposed to cultured BEAS-2B cells. Microarray analysis of the treated cells indicated a significant up-regulation of a number of genes associated with oxidative stress, including metallothionein, heme oxygenase-1, heat shock protein 70, and NAD(P)H dehydrogenase-1.

Identification and Characterization of Protein Encoded by orf382 as $\small{L}$-Threonine Dehydrogenase

  • Ma, Fei;Wang, Tianwen;Ma, Xingyuan;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.748-755
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    • 2014
  • In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).

Isolation and Characterization of a Formate Dehydrogenase cDNA in Poplar (Populus alba ${\times}$ P. glandulosa) (현사시나무에서 Formate Dehydrogenase cDNA의 분리와 특성 구명)

  • Bae, Eun-Kyung;Lee, Hyoshin;Lee, Jae-Soon;Choi, Young-Im;Yoon, Seo-Kyung;Eo, Soo Hyung
    • Journal of Korean Society of Forest Science
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    • v.102 no.3
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    • pp.331-337
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    • 2013
  • Formate dehydrogenase (FDH), catalyzing the oxidation of the formate ion to carbon dioxide, is known as the stress protein in response to drought, low temperature and pathogen infection. To study the functions of FDH in poplar (Populus alba ${\times}$ P. glandulosa), we isolated a FDH cDNA (PagFDH1) and examined its expressional characteristics. The PagFDH1 is 1,499 base pairs long and encodes a putative 388 amino acid protein with an expected molecular mass of 42.5 kDa. The PagFDH1 protein has N-terminal mitochondria signal peptide and $NAD^+$ binding domain. Southern blot analysis indicated that a single copy of the PagFDH1 is present in the poplar genome. PagFDH1 is expressed highly in the suspension cells (especially in the lag and early exponential phases) and moderately in roots, flowers and leaves. ABA-mediated enhanced expression of PagFDH1 in response to drought and salt stress treatments indicates that the gene product could play an important role in the development of stress resistant trees.

Supplementation with psyllium seed husk reduces myocardial damage in a rat model of ischemia/reperfusion

  • Lim, Sun Ha;Lee, Jongwon
    • Nutrition Research and Practice
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    • v.13 no.3
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    • pp.205-213
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    • 2019
  • BACKGROUND/OBJECTIVES: Myocardial infarction (MI) is caused by extensive myocardial damage attributed to the occlusion of coronary arteries. Our previous study in a rat model of ischemia/reperfusion (I/R) demonstrated that administration of arabinoxylan (AX), comprising arabinose and xylose, protects against myocardial injury. In this study, we undertook to investigate whether psyllium seed husk (PSH), a safe dietary fiber containing a high level of AX (> 50%), also imparts protection against myocardial injury in the same rat model. MATERIALS/METHODS: Rats were fed diets supplemented with PSH (1, 10, or 100 mg/kg/d) for 3 d. The rats were then subjected to 30 min ischemia through ligation of the left anterior descending coronary artery, followed by 3 h reperfusion through release of the ligation. The hearts were harvested and cut into four slices. To assess infarct size (IS), an index representing heart damage, the slices were stained with 2,3,5-triphenyltetrazolium chloride (TTC). To elucidate underlying mechanisms, Western blotting was performed for the slices. RESULTS: Supplementation with 10 or 100 mg/kg/d of PSH significantly reduces the IS. PSH supplementation (100 mg/kg/d) tends to reduce caspase-3 generation and increase BCL-2/BAX ratio. PSH supplementation also upregulates the expression of nuclear factor erythroid 2-related factor 2 (NRF2), and its target genes including antioxidant enzymes such as glutathione S-transferase mu 2 (GSTM2) and superoxide dismutase 2 (SOD2). PSH supplementation upregulates some sirtuins ($NAD^+$-dependent deacetylases) including SIRT5 (a mitochondrial sirtuin) and SIRT6 and SIRT7 (nuclear sirtuins). Finally, PSH supplementation upregulates the expression of protein kinase A (PKA), and increases phosphorylated cAMP response element-binding protein (CREB) (pCREB), a target protein of PKA. CONCLUSIONS: The results from this study indicate that PSH consumption reduces myocardial I/R injury in rats by inhibiting the apoptotic cascades through modulation of gene expression of several genes located upstream of apoptosis. Therefore, we believe that PSH can be developed as a functional food that would be beneficial in the prevention of MI.

Occurrence of Apple Scar Skin viroid and Relative Quantity Analysis Using Real-time RT-PCR (Apple Scar Skin viroid 발생상황 및 Real-time RT-PCR을 이용한 상대정량 분석)

  • Kim, Dae-Hyun;Kim, Hyun-Ran;Heo, Seong;Kim, Se-Hee;Kim, Min-A;Shin, Il-Sheob;Kim, Jeong-Hee;Cho, Kang-Hee;Hwang, Jeong-Hwan
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.247-253
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    • 2010
  • Apple scar skin viroid (ASSVd) is one of the smallest viral pathogens infecting fruits, especially apple, and causes a significant damage to fruit trees. ASSVd usually induced the skin-dapple ring symptoms, but in 'Fuji' varieties, corked spot were occurred on the fruit skin in 2009. This new symptom will be of great helpful to diagnosis ASSVd in sight. ASSVd was surveyed in apple and pear from 2009 to 2010 in Korea, and ASSVd was identified in 20 out of 1,193 trees. The infection rate was 1.7%. To screen the infectivity of ASSVd among apple cultivars, real-time RT-PCR was applied followed by designing of ASSVd specific primers based on highly conserved regions of several ASSVd isolates including Korean isolate. NADH dehydrogenase subunit 5 (nad 5) gene, which is mRNA of the mitochondrial gene, was used for internal control. In this study, ASSVd infected apples were classified into 12 groups depending on different symptoms and symptom severity (scaring, rusting or malformation). Taken together, this study suggested that real-time PCR analysis was more sensitive to detect the low copy of ASSVd on early viroid infected apple skins than regular RT-PCR method.

Protection by Chrysanthemum zawadskii extract from liver damage of mice caused by carbon tetrachloride is maybe mediated by modulation of QR activity

  • Seo, Ji-Yeon;Lim, Soon-Sung;Park, Ji-A;Lim, Ji-Sun;Kim, Hyo-Jung;Kang, Hui-Jung;YoonPark, Jung-Han;Kim, Jong-Sang
    • Nutrition Research and Practice
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    • v.4 no.2
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    • pp.93-98
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    • 2010
  • Our previous study demonstrated that methanolic extract of Chrysanthemum zawadskii Herbich var. latilobum Kitamura (Compositae) has the potential to induce detoxifying enzymes such as NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) (NQO1, QR) and glutathione S-transferase (GST). In this study we further fractionated methanolic extract of Chrysanthemum zawadskii and investigated the detoxifying enzyme-inducing potential of each fraction. The fraction (CZ-6) shown the highest QR-inducing activity was found to contain (+)-(3S,4S,5R,8S)-(E)-8-acetoxy-4-hydroxy-3-isovaleroyloxy-2-(hexa-2,4-diynyliden)-1,6-dioxaspiro [4,5] decane and increased QR enzyme activity in a dose-dependent manner. Furthermore, CZ-6 fraction caused a dose-dependent enhancement of luciferase activity in HepG2-C8 cells generated by stably transfecting antioxidant response element-luciferase gene construct, suggesting that it induces antioxidant/detoxifying enzymes through antioxidant response element (ARE)-mediated transcriptional activation of the relevant genes. Although CZ-6 fraction failed to induce hepatic QR in mice over the control, it restored QR activity suppressed by $CCl_4$ treatment to the control level. Hepatic injury induced by $CCl_4$ was also slightly protected by pretreatment with CZ-6. In conclusion, although CZ-6 fractionated from methanolic extract of Chrysanthemum zawadskii did not cause a significant QR induction in mice organs such as liver, kidney, and stomach, it showed protective effect from liver damage caused by $CCl_4$.