• Title/Summary/Keyword: RNA quality control

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Correlation between Heart-type Fatty Acid-binding Protein Gene Polymorphism and mRNA Expression with Intramuscular Fat in Baicheng-oil Chicken

  • Wang, Yong;He, Jianzhong;Yang, Wenxuan;Muhantay, Gemenggul;Chen, Ying;Xing, Jinming;Liu, Jianzhu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1380-1387
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    • 2015
  • This study aims to determine the polymorphism and mRNA expression pattern of the heart-type fatty acid-binding protein (H-FABP) gene and their association with intramuscular fat (IMF) content in the breast and leg muscles of Baicheng oil chicken (BOC). A total of 720 chickens, including 240 black Baicheng oil chicken (BBOC), 240 silky Baicheng oil chicken (SBOC), and 240 white Baicheng oil chicken (WBOC) were raised. Three genotypes of H-FABP gene second extron following AA, AB, and BB were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) strategy. The G939A site created AA genotype and G956A site created BB genotype. The content of IMF in AA genotype in breast muscle of BBOC was significantly higher than that of AB (p = 0.0176) and the genotype in leg muscle of WBOC was significantly higher than that of AB (p = 0.0145). The G939A site could be taken as genetic marker for higher IMF content selecting for breast muscle of BBOC and leg muscle of WBOC. The relative mRNA expression of H-FABP was measured by real-time PCR at 30, 60, 90, and 120 d. The IMF content significantly increased with age in both muscles. The mRNA expression level of H-FABP significantly decreased with age in both muscles of the three types of chickens. Moreover, a significant negative correlation between H-FABP abundance and IMF content in the leg muscles of WBOC (p = 0.035) was observed. The mRNA expression of H-FABP negatively correlated with the IMF content in both breast and leg muscles of BOC sat slaughter time.

Sequence and Expression Analysis of c-fos Proto-oncogene in Korean Cattle (HANWOO) (한우 c-fos 유전자의 염기서열 및 발현분석)

  • Yu, S.L.;Chung, H.J.;Jung, K.C.;Lee, J.H.;Cho, K.W.;Choi, J.G.;Na, K.J.;Sang, B.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.891-900
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    • 2003
  • Cellular FOS(c-fos) protein is a transcription factor that forms heterodimers mostly with c-jun family and stimulates the transcription of genes containing AP-1 regulatory elements. This c-fos expression can control growth and differentiation of various precursor cells including myoblasts. The controls by c-fos gene have been identified for affecting skeletal muscle fiber traits which are the key determinants of meat quality in pigs. As a first step for identifying the relationship between c-fos gene and meat quality traits in cattle, we fully sequenced 1,443 bp of Hanwoo c-fos mRNA and analyzed expression patterns from various organs and muscle tissues. The sequence identities of Hanwoo c-fos with that of human, pig and mouse showed 89.8%, 93.3% and 87%, respectively. Analyses of the northern blot showed high c-fos expressions were obtained in spleen and rib muscle from 7 organs and 9 different parts of muscles investigated. These results presented here can be used as a valuable marker for meat quality related traits in cattle with further verification.

Effects of enzymolysis and fermentation of Chinese herbal medicines on serum component, egg production, and hormone receptor expression in laying hens

  • Mei Hong Jiang;Tao Zhang;Qing Ming Wang;Jin Shan Ge;Lu Lu Sun;Meng Qi Li;Qi Yuan Miao;Yuan Zhao Zhu
    • Animal Bioscience
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    • v.37 no.1
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    • pp.95-104
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    • 2024
  • Objective: In the present study, we aimed to investigate the effects of enzymolysis fermentation of Chinese herbal medicines (CHMs) on egg production performance, egg quality, lipid metabolism, serum reproductive hormone levels, and the mRNA expression of the ovarian hormone receptor of laying hens in the late-laying stage. Methods: A total of 360 Hy-Line Brown laying hens (age, 390 days) were randomly categorized into four groups. Hens in the control (C) group were fed a basic diet devoid of CHMs, the crushed CHM (CT), fermented CHM (FC), and enzymatically fermented CHM (EFT) groups received diets containing 2% crushed CHM, 2% fermented CHM, and 2% enzymatically fermented CHM, respectively. Results: Compared with crushed CHM, the acid detergent fiber, total flavonoids, and total saponins contents of fermented CHM showed improvement (p<0.05); furthermore, the neutral and acid detergent fiber, total flavonoids, and total saponins contents of enzymatically fermented CHM improved (p<0.05). At 5 to 8 weeks, hens in the FC and EFT groups showed increased laying rates, haugh unit, albumin height, yolk color, shell thickness, and shell strength compared with those in the C group (p<0.05). Compared with the FC group, the laying rate, albumin height, and Shell thickness in the EFT group was increased (p<0.05). Compared with the C, CT, and FC groups, the EFT group showed reduced serum total cholesterol and increased serum luteinizing hormone levels and mRNA expressions of follicle stimulating hormone receptor and luteinizing hormone receptor (p<0.05). Conclusion: These results indicated that the ETF group improved the laying rate and egg quality and regulated the lipid metabolism in aged hens. The mechanism underlying this effect was likely related to cell wall degradation of CHM and increased serum levels of luteinizing hormone and mRNA expression of the ovarian hormone receptor.

Effect of Dietary Fermented Garlic Solution on Performance, Egg Quality and Blood Composition in Finishing Period of Laying Hens (마늘발효액의 첨가 급여가 산란계의 산란말기 생산성, 계란 품질 및 혈액성상에 미치는 영향)

  • Lim, Chun Ik;Kang, Chang Won;Chun, Hyeon Soo;Choi, Ho Sung;Ryu, Kyeong Seon
    • Korean Journal of Poultry Science
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    • v.45 no.3
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    • pp.201-207
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    • 2018
  • An experiment was conducted to investigate the effect of fermented garlic solution (FGS) on the performance, egg quality and blood profiles of laying hens in the finishing period. In total, 432 Lohmann Brown hens aged 79 weeks were equally distributed into four dietary treatments with six replicate. Hens were fed the basal diet containing 2,750 kcal/kg of ME and 16% of CP, which was supplemented with either 0% (control), 0.05%, 0.10% and 0.20% FGS from 79 to 83 weeks old. Laying performance, egg quality, yolk fatty acids and serum characteristics were analyzed at the end of experiment. Egg production and feed conversion was numerically improved in FGS supplementation treatments compared to those in the control, but were not statistically different. The albumen height and Haugh unit showed significant increase (P<0.05) in the FGS supplementation groups. The concentration of saturated fatty acid decreased in the yolks of birds fed FGS (P<0.01), whereas the unsaturated fatty acid (UFA) and mono-UFA contents were significantly higher (P<0.01) in those treatments than in the control. Significantly lower natural fat and cholesterol in serum were observed in birds fed the 0.20% FGS supplementation diet (P<0.01). However, the high-density lipoprotein (HDL) cholesterol increased in both the 0.10% and 0.20% FGS supplementation groups. In addition, interleukin-2 mRNA and CD4+/CD8+ level in serum which were cellular immunity indicators showed statistical differences (P<0.01) among treatments and a higher concentration in the 0.10% and 0.20% FGS groups than in the control. Thus, it can be concluded that dietary supplementation of FGS improved egg quality and stimulated immune response in mature laying hens.

Light modulates the transcriptomic accumulation of anthocyanin biosynthetic pathway genes in red and white grapes

  • Puspa Raj Poudel ;Kazuya Koyama ;Nami Goto-Yamamoto
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.292-2999
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    • 2022
  • Anthocyanin, an important component in the grape berry skin, strongly affects grape quality. The transcription factors VvMYBA1 and VvMYBA2 (VvMYBA1/2) control anthocyanin biosynthesis. In addition, cultivation and environmental factors, such as light, influence anthocyanin accumulation. The present study aimed to clarify the effect of shading (reduced light condition) on the transcriptomic regulation of anthocyanin biosynthesis using a red-wine grape cultivar, Vitis vinifera 'Pinot Noir', and its white mutant, 'Pinot Blanc', caused by the deletion of the red allele of VvMYBA1/2. The grape berry skins were analyzed for anthocyanin content and global gene transcription accumulation. The microarray data were later validated by quantitative real-time PCR. A decisive influence of VvMYBA1/2 on the expression of an anthocyanin-specific gene, UDP glucose: flavonoid 3-O-glucosyltransferase, was observed as expected. In contrast, upstream genes of the pathway, which are shared by other flavonoids, were also expressed in 'Pinot Blanc', and the mRNA levels of some of these genes decreased in both cultivars on shading. Thus, the involvement of light-sensitive transcription factor(s) other than VvMYBA1/2 was suggested for the expression control of the upstream genes of the anthocyanin biosynthetic pathway. Furthermore, it was suggested that the effects of these factors are different among isogenes.

Biological Control of Lettuce Sclerotinia Rot by Bacillus subtilis GG95 (길항미생물 Bacillus subtilis GG95를 이용한 상추 균핵병의 생물학적 방제)

  • Lee, Hyun-Ju;Kim, Jin-Young;Lee, Jin-Gu;Hong, Soon-Sung
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.225-230
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    • 2014
  • Sclerotinia sclerotiorum, a plant pathogenic fungus, can cause serious yield and quality losses in the winter lettuce field. For biological control of S. sclerotiorum, soil-born microorganisms that inhibit the mycelia growth of S. sclerotiorum and Fusarium oxysporum were isolated from diseased soil. Among the isolates, bacterial isolate, GG95, which was identified as Bacillus subtilis according to the morphological, physiological characteristics and by 16S rRNA similarity, showed the highest level of inhibitory activity. The growth conditions for B. subtilis GG95 were optimized in TSB media (pH 7) by culturing at $28^{\circ}C$ for 24 hrs. Maltose or fructose and peptone were selected as the best carbon and nitrogen sources, respectively. Greenhouse experiment was performed to test effectiveness of B. subtilis GG95 in the control sclerotinia rot. Drench application ($1{\times}10^8cfu/mL$, 3 times) of the bacterial culture broth to lettuce showed an effectiveness value of 88%, suggesting that B. subtilis GG95 would be a promising biocontrol agent for control of sclerotinia rot.

Effects of Dietary Thiazolidinedione Supplementation on Growth Performance, Intramuscular Fat and Related Genes mRNA Abundance in the Longissimus Dorsi Muscle of Finishing Pigs

  • Chen, X.;Feng, Y.;Yang, W.J.;Shu, G.;Jiang, Q.Y.;Wang, X.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.7
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    • pp.1012-1020
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    • 2013
  • The objective of this study was to investigate the effect of dietary supplementation with thiazolidinedione (TZD) on growth performance and meat quality of finishing pigs. In Experiment 1, 80 castrated finishing pigs (Large White${\times}$Landrace, BW = 54.34 kg) were randomly assigned to 2 treatments with 5 replicates of 8 pigs each. The experimental pigs in the 2 groups were respectively fed with a diet with or without a TZD supplementation (15 mg/kg). In Experiment 2, 80 castrated finishing pigs (Large White${\times}$Landrace, BW = 71.46 kg) were divided into 2 treatments as designed in Experiment 1, moreover, carcass evaluations were performed. The results from Experiment 1 showed that TZD supplementation could significantly decreased the average daily feed intake (ADFI) (p<0.05) during 0 to 28 d, without impairing the average daily gain (ADG) (p>0.05). In Experiment 2, the ADG was significantly increased by TZD supplementation during 14 to 28 d and 0 to 28 d (p<0.05) and the feed:gain ratio (F:G) was significantly decreased by TZD supplementation during 0 to 28 d (p<0.05). Compared with the control group, TZD group had significantly higher serum triglyceride (TG) concentration at 28h and serum high-density lipoprotein (HDL) levels at 14 d (p<0.05). Moreover, there was an apparent improvement in the marbling score (p<0.10) and intramuscular fat (IMF) content (p<0.10) of the longissimus dorsi muscle in pigs treated by TZD supplementation. Real-time RT-PCR analyses demonstrated that pigs of TZD group had higher mRNA abundance of $PPAR{\gamma}$ coactivator 1 (PGC-1) (p<0.05) and fatty acid-binding protein 3 (FABP3) (p<0.05) than pigs of control group. Taken together, these results suggested that dietary TZD supplementation could improve growth performance and increase the IMF content of finishing pigs through regulating the serum parameters and genes mRNA abundance involved in fat metabolism.

Effect of dietary supplementation with Spirulina on the expressions of AANAT, ADRB3, BTG2 and FASN genes in the subcutaneous adipose and Longissimus dorsi muscle tissues of purebred and crossbred Australian sheep

  • Kashani, Arash;Holman, Benjamin William Behrens;Nichols, Peter David;Malau-Aduli, Aduli Enoch Othniel
    • Journal of Animal Science and Technology
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    • v.57 no.3
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    • pp.8.1-8.8
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    • 2015
  • Background: The demand for healthy, lean and consistent meat products containing low saturated fatty acid content and high quality polyunsaturated fatty acids (PUFA), especially long-chain (${\geq}C_{20}$) omega-3 PUFA, has increased in recent times. Fat deposition is altered by both the genetic background and dietary supplements, and this study aimed to assess the effect of dietary Spirulina supplementation levels on the mRNA expression patterns of genes controlling lipid metabolism in the subcutaneous adipose tissue (SAT) and Longissimus dorsi (ld) muscle of Australian crossbred sheep. Methods: Twenty-four weaned lambs belonging to four breeds under the same management conditions were maintained on ryegrass pasture and fed three levels of Spirulina supplement (control, low and high). In terms of nutrient composition, Spirulina is a nutrient-rich supplement that contains all essential amino acids, vitamins and minerals. It also is a rich source of carotenoids and fatty acids, especially gamma-linolenic acid (GLA) that infer health benefits. After slaughter, subcutaneous adipose tissue (SAT) and ld samples were subjected to mRNA extraction and reverse transcription using quantitative polymerase chain reaction (RT-qPCR) to assess the mRNA expression levels of the Aralkylamine N-acetyltransferase (AANAT), Adrenergic beta-3 receptor (ADRB3), B-cell translocation gene 2 (BTG2) and Fatty acid synthase (FASN) genes, which are associated with lipid metabolism. Results: Both low and high Spirulina supplementation levels strongly up-regulated the transcription of all the selected genes in both SAT and ld tissues (mostly in the subcutaneous adipose), but sheep breed and sex did not influence the gene expression patterns in these tissues. Conclusions: The evidence indicates that high Spirulina supplementation level resulted in a decrease in intramuscular fat content in Australian purebred and crossbred sheep due to the enhanced production of melatonin in sheep muscle tissues and strong up-regulation of mRNA expression of BTG2 in SAT which negatively affected fat deposition. In contrast, low Spirulina supplementation level strongly up-regulated the ADRB3 and FASN genes responsible for fat production. These findings are consistent with the observed phenotypic data suggesting that low Spirulina supplementation level can increase lamb production, with higher long-chain PUFA content.

Chronic persistent post-surgical pain following staging laparotomy for carcinoma of ovary and its relationship to signal transduction genes

  • Saxena, Ashok Kumar;Chilkoti, Geetanjali T;Chopra, Anand K;Banerjee, Basu Dev;Sharma, Tusha
    • The Korean Journal of Pain
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    • v.29 no.4
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    • pp.239-248
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    • 2016
  • Background: The present study was undertaken to evaluate the incidence of chronic persistent post-surgical pain (CPPP) and the role of signal transduction genes in patients undergoing staging laparotomy for carcinoma ovary. Methods: The present observational study was undertaken following institutional ethical committee approval and informed consent from all the participants. A total 21 patients of ASA grade I to III with age 20-70 years, scheduled for elective staging laparotomy for carcinoma ovary were included. Patients were excluded if had other causes of pain, cognitive dysfunction or chronic neurological disorders. Statistical analysis of pool data was done using SPSS version-17. For various scales like GPE, PDQ, NPSI, the visual analogue scale (VAS), global perceived effect (GPE), the pain DETECT questionnaire (PDQ), and neuropathic pain symptoms inventory (NPSI), one factor repaeted measure ANOVA applied with simple contrast with baseline as on post-operative day 1 (considered as reference and compared with subsequent time-interval), and the P values were adjusted according to "Bonferroni adjustments". In patients with CPPP, the ${\Delta}ct$ values of mRNA expressions of genes at the end of postoperative day 90 were compared with the baseline control values by one factor repeated ANOVA. P value < 0.005 significant. Results: The present study demonstrates 38.1% (8 out of 21 patients) incidence of CPPP. The functional status and quality of life as were observed to be significantly diminished in all patients with chronic pain. An up-regulation in the mRNA expression of signal transduction and a positive correlation was noted between the mRNA expression of signal transduction genes and VAS score in all patients with CPPP at the end of postoperative day 90. Conclusions: The reported incidence of CPPP in patients with carcinoma ovary was 38.1%. An up-regulation and positive correlation between mRNA expression of signal transduction genes and VAS score depicts its potential role in the pathogenesis of CPPP.

Inhibition of mitoNEET induces Pink1-Parkin-mediated mitophagy

  • Lee, Seunghee;Lee, Sangguk;Lee, Seon-Jin;Chung, Su Wol
    • BMB Reports
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    • v.55 no.7
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    • pp.354-359
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    • 2022
  • MitoNEET, a mitochondrial outer membrane protein containing the Asn-Glu-Glu-Thr (NEET) sequence, controls the formation of intermitochondrial junctions and confers autophagy resistance. Moreover, mitoNEET as a mitochondrial substrate undergoes ubiquitination by activated Parkin during the initiation of mitophagy. Therefore, mitoNEET is linked to the regulation of autophagy and mitophagy. Mitophagy is the selective removal of the damaged or unnecessary mitochondria, which is crucial to sustaining mitochondrial quality control. In numerous human diseases, the accumulation of damaged mitochondria by impaired mitophagy has been observed. However, the therapeutic strategy targeting of mitoNEET as a mitophagy-enhancing mediator requires further research. Herein, we confirmed that mitophagy is indeed activated by mitoNEET inhibition. CCCP (carbonyl cyanide m-chlorophenyl hydrazone), which leads to mitochondrial depolarization, induces mitochondrial dysfunction and superoxide production. This, in turn, contributes to the induction of mitophagy; mitoNEET protein levels were initially increased before an increase in LC3-II protein following CCCP treatment. Pharmacological inhibition of mitoNEET using mitoNEET Ligand-1 (NL-1) promoted accumulation of Pink1 and Parkin, which are mitophagy-associated proteins, and activation of mitochondria-lysosome crosstalk, in comparison to CCCP alone. Inhibition of mitoNEET using NL-1, or mitoNEET shRNA transfected into RAW264.7 cells, abrogated CCCP-induced ROS and mitochondrial cell death; additionally, it activated the expression of PGC-1α and SOD2, regulators of oxidative metabolism. In particular, the increase in PGC-1α, which is a major regulator of mitochondrial biogenesis, promotes mitochondrial quality control. These results indicated that mitoNEET is a potential therapeutic target in numerous human diseases to enhance mitophagy and protect cells by maintaining a network of healthy mitochondria.