• Title/Summary/Keyword: Succinate

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Association Analysis of Myosin Heavy-chain Genes mRNA Transcription with the Corresponding Proteins Expression of Longissimus Muscle in Growing Pigs

  • Men, X.M.;Deng, B.;Tao, X.;Qi, K.K.;Xu, Zi Wei
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.457-463
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    • 2016
  • The goal of this work was to investigate the correlations between MyHC mRNA transcription and their corresponding protein expressions in porcine longissimus muscle (LM) during postnatal growth of pigs. Five DLY ($Duroc{\times}Landrace{\times}Yorkshire$) crossbred pigs were selected, slaughtered and sampled at postnatal 7, 30, 60, 120, and 180 days, respectively. Each muscle was subjected to quantity MyHCs protein contents through an indirect enzyme-linked immunosorbent assay (ELISA), to quantity myosin heavy-chains (MyHCs) mRNA abundances using real-time polymerase chain reaction. We calculated the proportion (%) of each MyHC to total of four MyHC for two levels, respectively. Moreover, the activities of several key energy metabolism enzymes were determined in LM. The result showed that mRNA transcription and protein expression of MyHC I, IIa, IIx and IIb in LM all presented some obvious changes with postnatal aging of pigs, especially at the early stage after birth, and their mRNA transcriptions were easy to be influenced than their protein expressions. The relative proportion of each MyHC mRNA was significantly positively related to that of its corresponding protein (p<0.01), and MyHC I mRNA proportion was positively correlated with creatine kinase (CK), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) activities (p<0.05). These data suggested that MyHC mRNA transcription can be used to reflect MyHC expression, metabolism property and adaptive plasticity of porcine skeletal muscles, and MyHC mRNA composition could be a molecular index reflecting muscle fiber type characteristics.

Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (Anser anser domestica) Gene Expression

  • Ji, Hong;Wang, Jianfa;Liu, Juxiong;Guo, Jingru;Wang, Zhongwei;Zhang, Xu;Guo, Li;Yang, Huanmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.3
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    • pp.423-432
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    • 2013
  • Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.

1H NMR-based metabolite profiling of diet-induced obesity in a mouse mode

  • Jung, Jee-Youn;Kim, Il-Yong;Kim, Yo-Na;Kim, Jin-Sup;Shin, Jae-Hoon;Jang, Zi-Hey;Lee, Ho-Sub;Hwang, Geum-Sook;Seong, Je-Kyung
    • BMB Reports
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    • v.45 no.7
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    • pp.419-424
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    • 2012
  • High-fat diets (HFD) and high-carbohydrate diets (HCD)-induced obesity through different pathways, but the metabolic differences between these diets are not fully understood. Therefore, we applied proton nuclear magnetic resonance ($^1H$ NMR)-based metabolomics to compare the metabolic patterns between C57BL/6 mice fed HCD and those fed HFD. Principal component analysis derived from $^1H$ NMR spectra of urine showed a clear separation between the HCD and HFD groups. Based on the changes in urinary metabolites, the slow rate of weight gain in mice fed the HCD related to activation of the tricarboxylic acid cycle (resulting in increased levels of citrate and succinate in HCD mice), while the HFD affected nicotinamide metabolism (increased levels of 1-methylnicotineamide, nicotinamide-N-oxide in HFD mice), which leads to systemic oxidative stress. In addition, perturbation of gut microflora metabolism was also related to different metabolic patterns of those two diets. These findings demonstrate that $^1H$ NMR-based metabolomics can identify diet-dependent perturbations in biological pathways.

Mass Cultivation and Secondary Metabolite Analysis of Rhodobacter capsulatus PS-2 (광합성세균 Rhodobater capsulatus PS-2의 대량배양 최적화 및 대사산물 분석)

  • Bong, Ki Moon;Kim, Jong Min;Yoo, Jae-Hong;Park, In Chul;Lee, Chul Won;Kim, Pyoung Il
    • KSBB Journal
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    • v.31 no.3
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    • pp.158-164
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    • 2016
  • Plant growth promoting (PGP) hormones, which are produced in a small quantity by bacteria, affect in plant growth and development. PGPs play an important role on the crop productivity in agricultural field. In this study, a photosynthetic bacterial strain producing the PGP was isolated from paddy soil. Bacterial isolate was gram negative, rod-shaped and motility positive. From the 16s rRNA gene sequence analysis, the isolate was identified as Rhodobacter capsulatus PS-2. The mass cultivation of R. capsulatus PS-2 was optimized by considering of the carbon, nitrogen and inorganic salt sources. Optimal medium composition was determined as Na-succinate 4.5 g, yeast extract 5 g, $K_2HPO_4$ 1 g, $MgSO_4$ 5 g, per liter. From the result of 500 L fermentation for 2 days using the optimal medium, the viable cells were $8.7{\times}10^9cfu/mL$. R. capsulatus PS-2 strain produced the carotenoid and indole-3-acetic acid (IAA). The carotenoid extraction and quantitative analysis were performed by HCl-assisting method. Total carotenoid contents from R. capsulatus PS-2 culture broth were measured as $7.02{\pm}0.04$ and $6.93{\pm}0.05mg/L$ under photoheterotrophic and chemoheterotrophic conditions, respectively. To measure the productivity of IAA, colorimetric method was employed using Salkowski reagent at optical density 535 nm. The results showed that the highest content of IAA was $197.44{\pm}5.92mg/L$ in the optimal medium supplemented with 0.3% tryptophan.

Evaluation of Candidate Housekeeping Genes for the Normalization of RT-qPCR Analysis using Developing Embryos and Prolarvae in Russian Sturgeon Acipenser gueldenstaedtii (러시아 철갑상어(Acipenser gueldenstaedtii) 발생 시료의 RT-qPCR 분석을 위한 내재 대조군 유전자의 선정)

  • Nam, Yoon Kwon;Lee, Sang Yoon;Kim, Eun Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.1
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    • pp.95-106
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    • 2018
  • To evaluate appropriate reference genes for the normalization of quantitative reverse transcription PCR (RT-qPCR) data with embryonic and larval samples from Russian sturgeon Acipenser gueldenstaedtii, the expression stability of eight candidate housekeeping genes, including beta-actin (ACTB), elongation factor-1A (EF1A), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone 2A (H2A), ribosomal protein L5 (RPL5), ribosomal protein L7 (RPL7), succinate dehydrogenase (SDHA), and ubiquitin-conjugating enzyme E2 (UBE2A), were tested using embryonic samples from 12 developmental stages and larval samples from 11 ontogenic stages. Based on the stability rankings from three statistic software packages, geNorm, NormFinder, and BestKeeper, the expression stability of the embryonic subset was ranked as UBE2A>H2A>SDHA>GAPDH>RPL5>EF1A>ACTB>RPL7. On the other hand, the ranking in the larval subset was determined as UBE2A>GAPDH>SDHA>RPL5>RPL7>H2A>EF1A>AC TB. When the two subsets were combined, the overall ranking was UBE2A>SDHA>H2A>RPL5>GAPDH>EF1A>ACTB>RPL7. Taken together, our data suggest that UBE2A and SDHA are recommended as suitable references for developmental and ontogenic samples of this sturgeon species, whereas traditional housekeepers such as ACTB and GAPDH may not be suitable candidates.

Factors Relating to Induced Systemic Resistance in Watermelon by Plant Growth-Promoting Pseudomonas spp.

  • Lee, Yong-Hoon;Lee, Wang-Hyu;Lee, Du-Ku;Shim, Hyeong-Kwon
    • The Plant Pathology Journal
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    • v.17 no.3
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    • pp.174-179
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    • 2001
  • The plant growth-promoting Pseudomonas strains, WR8-3 (Pseudomonas fluorescens), WR9-11 (Pseudomonas sp.) and WR9-16 (P.putida), which induced resistance systematically in watermelon to gummy stem rot were investigated on their induced systemic resistance(ISR)-related characteristics. The pyoverdine production was repressed in the standard succinate medium by increasing the concentration of $\textrm{FeCL}_3$. But the iron-binding ability on chrome azurol S agar media (CAS) was observed only in the strains, WR8-3 and WR9-16. When the two strains were mutated, the resulting iron-binding siderophore-negative mutants, WR8-3m and WR 9-16m, failed to promote the growth of watermelon and to induce resistance. The strains, WR8-3 and WR 9-16, slightly inhibited the growth of Didymella bryoniae at a low concentration of $\textrm{FeCL}_3$ on Kong's medium B, but not to exert control dffect. The strain WR9-11 showed antagonism in the concentration of $\textrm{FeCL}_3$ from 0 to $1,000\mu\textrm{M}$. When the crude lipoplysaccharide of each strain was treated in the rhizosphere of watermelon, mean lesion area was similar to that of the untreated control. The strains, WR9-11 and WR9-16 produced some level of hydrogen cyanide (HCN). Salicylic acid production was not detected in all of the strains.

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Effects of Cyclophosphamide in the Epididymis of the Rat I. Caput (Cyclophosphamide가 흰쥐의 부정소에 미치는 영향 I. 두부)

  • Cho, Kwang-Phil;Kim, Saeng-Gon;Jung, Hae-Man;Kim, Jeong-Sang;Kim, Young-Gon;Rho, Young-Bok
    • Applied Microscopy
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    • v.22 no.1
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    • pp.89-102
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    • 1992
  • This research was undertaken to determine the effects of the anticancer and immunosuppressive drug cyclophosphamide (CP) on the epididymal caput of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group, 5 weeks group were treated with saline (control group) or CP at doses of 20 mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymis, the mitochondrial outer and inner membranes were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also swollen, and the number of Golgi vesicles were increased, respectively. It is suggested that treatment with CP alters the specific cell organelles in the epididymis. CP caused changes in protein concentrations in caput of epididymis after CP treatment. Total proteins of 32 to 37 species such as lactate dehydrogenase, carnitine acetyltransferase and succinate dehydrogenase were expressed in the caput fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymis. In contrast to the control group, in particular carnitine acetyltransferase and the other 9 proteins in the caput fluid were decreased or disappeared, respectively, whereas lactate dehydrogenase and the other 5 proteins in the caput fluid were increased or synthesized, respectively. The other proteins are not showed distinctive difference. These alterations could be direct mediated by toxic effects of the drug on the epithelium or be secondary to changes in the spermatozoa as a result of the CP treatment.

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Effects of Mercury on the Differentiation Cerebral Neuron of Chick Embry (III) (수은이 계배 대뇌의 신경세포 분화에 미치는 영향(III))

  • Jeong, Hae-Man;Kim, Saeng-Gon;Cho, Kwang-Phil
    • Applied Microscopy
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    • v.27 no.1
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    • pp.87-100
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    • 1997
  • To investigate the effects of mercuric chloride $(HgCl_2)$ on the differentiation of the cerebral neuron of chick embryo 10 days, the ultrastructural changes in nerve cells injected with a various doses of mercuric chloride were observed with transmission electron microscope. The enzyme activity of the some dehydrogenases, cerebral proteins and adenosine triphosphate (ATP) were also analyzed. The results obtained are as follows; The ultrastructural changes in 1.0 mg-injected group, the nuclear membranes were irregular, outer of mitochondria membrances dispressioned, their cristae were destroyed. In 2.0 mg-injected group, the nuclear envelops were destroyed and divided, were not observed organelle except of few ribosome, the RER and mitochondria. The number of polypeptide bands were separated by SDS-PAGE in the normal group were 38 bands. According to the in creased dose of mercuric chloride, contends of the bands were increased in 4 bands, but were decreased in 1 band. The activities of dehydrogenases were declined by increasing the dose of mercuric chloride. Lactate dehydrogenase (LDH) activity fatted to 61% in 2.0 mg-injected group. Malate dehydrogenase (MDH) activity fatted to 90% in 1.0 mg-injected group, greatly to 76% in 2.0 mg-injected group. Succinate dehydrogenase (SDH) activity decreased to 79% in 1.0 mg-injected group and greatly to 62% in 2.0 mg-injected group. ATP content in 1.0 mg-injected group was almost near to the normal level, but it was increased greatly in 2.0 mg-injected group.

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Fishing power estimation of biodegradable traps in the East Sea (동해 생분해성 통발의 어획성능 평가)

  • Bae, Bong-Seong;An, Heui-Chun;Jeong, Eui-Cheol;Park, Hae-Hoon;Park, Seong-Wook;Park, Chang-Doo
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.46 no.4
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    • pp.292-301
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    • 2010
  • For an effective management of fisheries resources, it is very important that to make clean inhabitation environment and to preserve fisheries resources. The material which is mainly used as fishing gear in modern times, is polyethylene, polypropylene, polyamide, etc., chemical fiber. And lost fishing gears which are make of these, occur ghost fishing and ocean pollution. To solve these problem, we development biodegradable fishing trap using the polybutylene succinate (PBS). Developed traps are for red snow crab (Chionoecetes japonicus) and shrimp, major traps in the East Sea, and we carried out fishing research using two kind traps in the coastal sea of Ayajin-port (Goseong) to analysis fishing efficiency of PE trap and PBS trap. As a result for fishing experiment (year 2005-2006) of red snow crab trap, two kind traps were almost the same in catches and length composition. During a experiment, parts of meshes, used for over 1 year, were cut by biodegradation. As a result for fishing experiment (year 2007) of shrimp trap, northern shrimp (Pandalus eous), coonstripe shrimp (Pandalus hypsinotus) and morotoge shrimp (Pandalopsis japonica) were catched, and the almost is northern shrimp. Two kind traps were almost the same in catches and length composition. In accordance with these result, it is recommended that the developed traps are have to commercialized because the fishing powers of PE traps and PBS traps were same. But biodegradation speed is have to controled in consideration of ocean microorganism volume and traps life.

Taurine Regulates Mitochondrial Function During 7,12-Dimethyl Benz[a]anthracene Induced Experimental Mammary Carcinogenesis

  • Vanitha, Manickam Kalappan;Priya, Kalpana Deepa;Baskaran, Kuppusamy;Periyasamy, Kuppusamy;Saravanan, Dhravidamani;Venkateswari, Ramachandran;Mani, Balasundaram Revathi;Ilakkia, Aruldass;Selvaraj, Sundaramoorthy;Menaka, Rajendran;Geetha, Mahendran;Rashanthy, Nadarajah;Anandakumar, Pandi;Sakthisekaran, Dhanapal
    • Journal of Pharmacopuncture
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    • v.18 no.3
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    • pp.68-74
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    • 2015
  • Objectives: The present study was undertaken to determine the modulatory effect of taurine on the liver mitochondrial enzyme system with reference to mitochondrial lipid peroxidation (LPO), antioxidants, major tricarboxylic acid cycle enzymes, and electron transport chain enzymes during 7,12-dimethyl benz[a]anthracene (DMBA) induced breast cancer in Sprague-Dawley rats. Methods: Animals in which breast cancer had been induced by using DMBA (25 mg/kg body weight) showed an increase in mitochondrial LPO together with decreases in enzymic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST)), non-enzymic antioxidants (reduced glutathione (GSH), vitamin C, and vitamin E), in citric acid cycle enzymes (isocitrate dehydrogenase (ICDH), alpha ketoglutarate dehydrogenase (alpha KDH), succinate dehydrogenase (SDH) and malate dehydrogenase (MDH)), and in electron transport chain (ETC) complexes. Results: Taurine (100 mg/kg body weight) treatment decreased liver mitochondrial LPO and augmented the activities/levels of enzymic, and non-enzymic antioxidants, tricarboxylic acid cycle enzymes and ETC complexes. Conclusion: The results of our present study demonstrated the chemotherapeutic efficacy of taurine treatment for DMBA-induced breast carcinomas.