• Title/Summary/Keyword: synthetic gene

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Natural and synthetic pathogen associated molecular patterns modulate galectin expression in cow blood

  • Asiamah, Emmanuel Kwaku;Ekwemalor, Kingsley;Adjei-Fremah, Sarah;Osei, Bertha;Newman, Robert;Worku, Mulumebet
    • Journal of Animal Science and Technology
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    • v.61 no.5
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    • pp.245-253
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    • 2019
  • Pathogen-associated Molecular Patterns (PAMPs) are highly conserved structural motifs that are recognized by Pathogen Recognition receptors (PRRs) to initiate immune responses. Infection by these pathogens and the immune response to PAMPS such as lipopolysaccharide (LPS), Peptidoglycan (PGN), bacterial oligodeoxynucleotides [CpG oligodeoxynucleotides 2006 (CpG ODN2006) and CpG oligodeoxynucleotides 2216 (CpG ODN2216)], and viral RNA Polyinosinic-Polycytidylic Acid (Poly I:C), are associated with infectious and metabolic diseases in animals impacting health and production. It is established that PAMPs mediate the production of cytokines by binding to PRRs such as Toll-like receptors (TLR) on immune cells. Galectins (Gal) are carbohydrate-binding proteins that when expressed play essential roles in the resolution of infectious and metabolic diseases. Thus it is important to determine if the expression of galectin gene (LGALS) and Gal secretion in blood are affected by exposure to LPS and PGN, PolyI:C and bacterial CpG ODNs. LPS increased transcription of LGALS4 and 12 (2.5 and 2.02 folds respectively) and decreased secretion of Gal 4 (p < 0.05). PGN increased transcription of LGALS-1, -2, -3, -4, -7, and -12 (3.0, 2.3, 2.0, 4.1, 3.3, and 2.4 folds respectively) and secretion of Gal-8 and Gal-9 (p < 0.05). Poly I:C tended to increase the transcription of LGALS1, LGALS4, and LGALS8 (1.78, 1.88, and 1.73 folds respectively). Secretion of Gal-1, -3, -8 and nine were significantly increased in treated samples compared to control (p < 0.05). CpG ODN2006 did not cause any significant fold changes in LGALS transcription (FC < 2) but increased secretion of Gal-1, and-3 (p < 0.05) in plasma compared to control. Gal-4 was however reduced in plasma (p < 0.05). CpG ODN2216 increased transcription of LGALS1 and LGALS3 (3.8 and 1.6 folds respectively), but reduced LGALS2, LGALS4, LGALS7, and LGALS12 (-1.9, -2.0, -2.0 and; -2.7 folds respectively). Secretion of Gal-2 and -3 in plasma was increased compared to control (p < 0.05). Gal-4 secretion was reduced in plasma (p < 0.05). The results demonstrate that PAMPs differentially modulate galectin transcription and translation of galectins in cow blood.

Growth performance, carcass traits, muscle fiber characteristics and skeletal muscle mRNA abundance in hair lambs supplemented with ferulic acid

  • Pena-Torres, Edgar Fernando;Castillo-Salas, Candelario;Jimenez-Estrada, Ismael;Muhlia-Almazan, Adriana;Pena-Ramos, Etna Aida;Pinelli-Saavedra, Araceli;Avendano-Reyes, Leonel;Hinojosa-Rodriguez, Cindy;Valenzuela-Melendres, Martin;Macias-Cruz, Ulises;Gonzalez-Rios, Humberto
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.52-69
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    • 2022
  • Ferulic acid (FA) is a phytochemical with various bioactive properties. It has recently been proposed that due to its phytogenic action it can be used as an alternative growth promoter additive to synthetic compounds. The objective of the present study was to evaluate the growth performance, carcass traits, fiber characterization and skeletal muscle gene expression on hair-lambs supplemented with two doses of FA. Thirty-two male lambs (n = 8 per treatment) were individually housed during a 32 d feeding trial to evaluate the effect of FA (300 and 600 mg d-1) or zilpaterol hydrochloride (ZH; 6 mg d-1) on growth performance, and then slaughtered to evaluate the effects on carcass traits, and muscle fibers morphometry from Longissimus thoracis (LT) and mRNA abundance of β2-adrenergic receptor (β2-AR), MHC-I, MHC-IIX and IGF-I genes. FA increased final weight and average daily gain with respect to non-supplemented animals (p < 0.05). The ZH supplementation increased LT muscle area, with respect to FA doses and control (p < 0.05). Cross-sectional area (CSA) of oxidative fibers was larger with FA doses and ZH (p < 0.05). Feeding ZH increased mRNA abundance for β2-AR compared to FA and control (p < 0.05), and expression of MHC-I was affected by FA doses and ZH (p < 0.05). Overall, FA supplementation of male hair lambs enhanced productive variables due to skeletal muscle hypertrophy caused by MHC-I up-regulation. Results suggest that FA has the potential like a growth promoter in lambs.

Effects of amprolium hydrochloride on expression of drug metabolizing enzyme genes in olive flounder Paralichthys olivaceus (Amprolium hydrochloride가 넙치 Paralichthys olivaceus의 약물대사 유전자 발현에 미치는 영향)

  • Sang Hyup Park;Chang Han Kim;Jeong-wan Do;Hye-Sung Choi;Yi Kyung Kim
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.337-348
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    • 2023
  • This study was undertaken to evaluate the effect of amprolium hydrochloride on detoxification process of olive flounder Paralichthys olivaceus. A series of two experiment was performed based on the LD50 value obtained for amprolium. First, thirty flounder (average weight 230.27 g; average length 27.99 cm) was randomly allocated into five groups. Treatment was carried out using intra-muscular injection of amprolium at the dose levels of 4, 8, 16, and 32 mg/kg body weight. At 8, 24 and 48 h post injection, liver and kidney were collected for expression assay of drug metabolizing enzymes and pro-inflammatory cytokine genes. We found that the interleukin-1β (IL-1β) mRNA level were induced at 32 mg/kg and CYP1A genes showed the opposite pattern, while UDP-glucuronosyl-transferase (UGT1A7) and GST were significantly reduced in the liver. Moreover, the suppression of drug metabolizing enzymes and cytokine gene in the kidney was observed after treatment. Another treatment was carried out using intramuscular injection with 4, 8, 16, and 32 mg/kg and 60, 80, 100, 120 mg/kg body weight. At 6 days post injection, liver was collected. The IL-1β expression was markedly induced in the experimental group treated with 4 mg/kg. In addition, glutathione S-transferase (GST) mRNA level was higher in the group with 4 mg/kg. In conclusion, our data suggests that amprolium seem to cause direct or indirect physical, or biological toxicity of flounders, although this drug is considered one of the safest synthetic anticoccidial drugs of the livestock industry.

cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi (작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상)

  • Lee, Sang-Mong;Hwang, Jae-Sam;Park, Nam-Sook;Kim, Yong-Gyun;Kim, Keun-Ki;Son, Hong-Joo;Park, Hyun-Chul;Jin, Byung-Rae
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1193-1200
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    • 2010
  • The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

Comparison and Analysis between Human Breast Cancer Cells and Hepatoma Cells for the Effects of Xenobiotic Nuclear Receptors (Constitutive Androstane Receptor, Steroid and Xenobiotic Receptor, and Peroxisome-Proliferator-Activated Receptor γ ) on the Transcriptional Activity of Estrogen Receptor (유방암 세포와 간암세포에 있어서 에스트로겐 수용체의 전사조절기능에 대한 Xenobiotic 핵 수용체 (Constitutive Androstane Receptor, Steroid and Xenobiotic Receptor, Peroxisome-Proliferator-Activated Receptor γ )의 영향 비교분석)

  • 민계식
    • Journal of Life Science
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    • v.13 no.3
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    • pp.314-323
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    • 2003
  • The purpose of this study was to examine the effects of xenobiotic nuclear receptors, CAR, SXR, and PPAR${\gamma}$ on the transcriptional activity of estrogen receptor in human breast cancer cell lines and compare with those in human hepatoma cell line. Two different breast cancer cell lines, MCF-7 and MDA-MB-231 were cultured and effects of CAR, SXR, and PPAR${\gamma}$ on the ER-mediated transcriptional activation of synthetic (4ERE)-tk-luciferase reporter gene were analyzed. Consistent with the previous report, CAR significantly inhibited ER-mediated transactivation and SXR repressed modestly whereas the PPAR${\gamma}$ did not repress the ER-mediated transactivation. However, in breast cancer cells neither of the xenobiotic receptors repressed the ER-mediated transactivation. Instead, they tend to increase the transactivation depending on the cell type and xenobiotic nuclear receptors. In MCF-7, SXR but neither CAR nor PPAR${\gamma}$ slightly increased ER-mediated transactivation whereas in MDA-MB-231, CAR and PPAR${\gamma}$ but not SXR tend to increase the transactivation of the reporter gene. These results indicate that the effects of ER cross-talk by the CAR, SXR, and PPAR${\gamma}$ , are different in breast cancer cells from hepatoma cells. In conclusion, the transcriptional regulation by estrogen can involve different cross-talk interaction between estrogen receptor and xenobiotic nuclear receptors depending on the estrogen target cells.

Analysis of Stress Response of Domestic Chicken Breeds for the Development of a New Synthetic Parent Stock (국산 종계 개발을 위한 토종 계통들의 스트레스 반응 정도 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Ji Ae;Hong, Young Ho;Kim, Chong Dae
    • Korean Journal of Poultry Science
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    • v.42 no.2
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    • pp.157-167
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    • 2015
  • We compared the degrees of stress response of 12 domestic purebred chicken strains that have been bred at National Institute of Animal Science, RDA, Korea since 1980. As a physiological marker of stress response, the expression levels of heat shock protein (HSP)-70, HSP-$90{\alpha}$, HSP-$90{\beta}$, hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes and telomere length were measured by quantitative real-time polymerase chain reaction using the lymphocytes of 1,101 chickens. There was significant difference in HSP-70, HSP-$90{\alpha}$, HMGCR expression and telomere length among the strains. There was also significant difference in HSP-$90{\alpha}$, HSP-$90{\beta}$, and HMGCR expression between male and female chickens. Different age groups of chicken exhibited different expression levels of HSP-70, HSP-$90{\alpha}$ and telomere length. The results of the HSPs expression level suggested that, the strains of R, L and Y were highly resistant to stress, whereas the strains of S, O and W were susceptible to stress. Although the statistical differences in some of HSPs gene expression existed between genders, the HSP expression results varied in different strains that some opposed to the others, and there might be interaction between strains and genders, which conclude that there was no difference in stress response between male and female chickens. Moreover, despite of significant difference in some of HSPs expression level, it was considered that there was no difference in stress response between ages due to the inconsistent trends among HSP markers.

Biodegradation of Phenol by Comamonas testosteroni DWB-1-8 Isolated from the Activated Sludge of Textile Wastewater (섬유 폐수 활성 슬러지에서 분리한 Comamonas testosteroni의 생물학적 페놀 분해)

  • Kwon, Hae Jun;Choi, Doo Ho;Kim, Mi Gyeong;Kim, Dong-Hyun;Kim, Young Guk;Yoon, Hyeokjun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.30 no.2
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    • pp.156-161
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    • 2020
  • Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K2HPO4, 0.6% NaH2PO4, 0.1% NH4NO3, 0.015% MgSO4·7H2O, 0.001% FeSO4·7H2O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.

Effects of Hepcidin Hormone on the Gene Expression of Ferroportin and Divalent Metal Transporter 1 in Caco-2 Cells and J774 Cells (Caco-2 소장세포와 J774 대식세포에서 Hepcidin 호르몬이 철분 수송체 Ferroportin과 Divalent Metal Transporter 1의 유전자 발현에 미치는 영향)

  • Chae, Sun-Ju;Chung, Ja-Yong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.721-728
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    • 2008
  • Hepcidin is a peptide hormone produced by the liver, of which secretion is closely related to iron status in the body. However, little is known about the molecular mechanism(s) by which this peptide regulates body iron homeostasis. The purpose of this study was to determine the effects of hepcidin treatment within the physiological concentration range on the expressions of two different iron transporter proteins-ferroportin (FPN) and divalent metal transporter 1 (DMT1). Differentiated Caco-2 intestinal cells and macrophage J774 cells were treated with either synthetic hepcidin or hepcidin-rich fraction separated from human urine at the concentration of 10 nM and 100 nM for 24 hours. Results show that hepcidin treatment in differentiated Caco-2 cells or in J774 cells did not change the level of either FPN mRNA or DMT1 mRNA. On the other hand, hepcidin treatment at the dose of 100 nM significantly decreased the FPN protein levels and DMT1 protein levels in differentiated Caco-2 cells. Similarly, urinary hepcidin treatment (10 nM & 100 nM) also significantly decreased the levels of FPN and DMT1 proteins in J774 macrophage cells. These results showed that hepcidin might play an important role in the regulation of iron homeostasis by lowering the protein levels of iron transporter FPN and DMT1 both in enterocytes and in macrophage cells.

Detection of Auxotrophic Mutants form Valsa ceratosperma, the Causal Fungus of Apple Canker (사과나무 부란병균(腐爛病菌) Valsa ceratosperma에서의 Auxotrophic Mutants의 검출(檢出))

  • Hong, Yeon Gyu;Uhm, Jae Youl
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.119-126
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    • 1987
  • This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.

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Studies on Antioxidant, Anti-inflammatory and Whitening Effects of Oriental Herbal Extracts (Mix) including Eucommiae cortex (두충을 포함하는 한방추출물(Mix)의 항노화, 항염, 미백 효능 활성에 관한 연구)

  • Choi, Da Hee;Kim, Mi Ran;Kim, Min Young;Kim, Ho Hyun;Park, Sun-Young;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.37-47
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    • 2019
  • Recently, due to the increase in skin diseases caused by particulate matter, endocrine disruptor and environmental changes, the trend of development of cosmetic materials has been shifting to the more safe and effective ingredients based on natural materials rather than existing synthetic compounds like steroids and antihistamines. This study aimed to develop a new natural cosmetic materials using oriental herbs such as Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus. First, DPPH assay was performed to examine the antioxidative activity of the herbal extract (Mix) and 98.8% DPPH radical scavenging activity was confirmed at $400{\mu}g/mL$ concentration of it. In order to confirm the whitening efficacy of oriental herbal extracts(mix), the amount of melanin synthesized after stimulation of ${\alpha}-MSH$ with B16F10 cells was measured. Results showed that it was decreased to 27.1% comparing with the only ${\alpha}-MSH$ treated group, which confirmed the whitening efficacy. Also, both nitric oxide(NO) production and iNOS and COX-2 expression were significantly reduced in RAW264.7 macrophages activated by LPS in the presence of the extracts(Mix). The mRNA expression of the inflammatory cytokines such as $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ was also analyzed to confirm the inhibition effect of the extracts on inflammation. Finally, to confirm the enhancement of skin barrier function, the expression of claudin 1 gene, a tight junction protein, was observed using human keratinocyte HaCaT cells and increased as concentration dependent manner. From these results, it is concluded that the oriental herbal extracts(Mix) containing Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus is effective for antioxidant, anti-inflammation, skin whitening, and skin barrier and thus could be applied as a new natural cosmetic material.