• Title/Summary/Keyword: gene expression

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Gut Microbiome and Gut Immunity in Broiler Chickens Fed Allium hookeri Root Powder from Day 10 to 28 (육계 사료 내 삼채뿌리분말 첨가가 장내 미생물 및 장관면역에 미치는 영향)

  • Woonhak Ji;Inho Cho;Sang Seok Joo;Moongyeong Jung;Chae Won Lee;June Hyeok Yoon;Su Hyun An;Myunghoo Kim;Changsu Kong
    • Korean Journal of Poultry Science
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    • v.50 no.3
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    • pp.171-185
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    • 2023
  • This study was conducted to investigate the effects of supplementation of Allium hookeri (AH) root powder on the gut microbiome, immunity, and health in broiler chickens fed experimental diets from d 10 to 28. A total of 60 10-day-old Ross 308 broilers were weighed and assigned to two dietary treatments with 5 birds per cage in a randomized complete block design based on body weight. The two experimental diets consisted of a control diet based on corn-soybean meal and the control diet supplemented with 0.3% AH root powder. All birds were fed ad libitum with experimental diets and water for 18 d. At 28 d, two birds near the median weight from each cage were selected for cecal content and small intestinal tissue sample collection. The addition of AH changed the gut microbiome by increasing probiotic candidate beneficial bacteria such as Enterococcaceae, Lactobacillaceae, Limosilactobacillus, Cuneatibacter, and Ruminoccoides. Regarding gut immunity, the supplementation of AH resulted in changes in intestinal immune cells, including reduced CD3+CD4+ T cells, which are a type of helper T cell, in the small intestine of birds (P=0.049). Additionally, there was a tendency to increase the expression of antioxidant function-related gene such as GPX2 (P=0.060), but no significant changes were observed in cytokines such as IL1b, IL6, and IL10. Overall, the addition of AH root powder may have positive effects on the microbiome of the chickens. This may help promote gut health in broiler chickens at the age of d 10 to 28.

Comparison of Agrobacterium-mediated Transformation Efficiency in 43 Korean Wheat Cultivars (국내 밀 43개 품종에 대한 아그로박테리움 형질전환 효율성 검정)

  • Jae Yoon Kim;Geon Hee Lee;Ha Neul Lee;Do Yoon Hyun
    • Journal of Practical Agriculture & Fisheries Research
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    • v.25 no.4
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    • pp.138-147
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    • 2024
  • Agrobacterium-mediated transformation (AMT) is a method that allows for the stable integration of DNA fragments into the plant genome. Transgenic plants generated through AMT typically exhibit a lower copy number of the transgene compared to those induced by particle bombardment. Furthermore, AMT offers a straightforward and efficient approach for generating transgenic plants. While the transformation efficiency of wheat is comparatively lower than that of other monocot plants such as Rice (Oryza sativa L.) and Maize (Zea mays L.), the cultivars 'Bobwhites' and 'Fielder' are commonly employed for wheat transformation. To date, there have been no reported instances of successful development of transgenic plants using Korean wheat varieties through AMT. This study aims to assess the transformation efficiency of 43 Korean wheat cultivars using the GUS assay, with the goal of identifying suitable Korean wheat cultivars for AMT. The pCAMBIA1301 vector, carrying the β-glucuronidase (GUS) gene, was incorporated into Agrobacterium strain EH105. Following the inoculation of Agrobacterium into immature embryos, GUS assays were conducted 'Saeol', 'Jopum', and 'Jonong' showed 100% (the number of embryos showing GUS spots/the number of embryos used for AMT) among 43 cultivars. In addition, cultivars with more than 70% were 'Saekeumgang', 'Jojung', 'Tapdong', 'Anbaek', 'Dabun', 'Sugang', 'Keumgang', 'Jeokjung', 'Seodun', 'Joeun', 'Dajung', and 'Baekjung'. It seems that the 15 cultivars above showed the possibility of using AMT. On the other hand, 'Yeonbaek', 'Goso', 'Baekgang', and 'Johan' showed less than 20% and GUS spots were not observed in 'Gru', 'Gobun', 'Milseong', and 'Shinmichal-1'. This study explores transient GUS expression in Korean wheat cultivars seven days after AMT. The observed initial high efficiency of transient transformation suggests the potential for subsequent stable transformation efficiency. Korean wheat cultivars demonstrating elevated transient transformation efficiency could serve as promising candidates for the development of stable transgenic wheat.

Ethanol extract of Aster glehni exhibits anti-inflammatory and anti-oxidant effects in RAW 264.7 cells and Caenorhabditis elegans (섬쑥부쟁이 에탄올 추출물이 대식세포와 예쁜꼬마선충에서의 항염증 및 항산화 효과 )

  • Mi-Kyung Seo;Han-Na Chu;Da-Bin Lee;Haeng-Ran Kim;In-Seon Hwang;Yong-Jin Jeong;Sung-Ran Yoon;Seok-Seong Kang;Kyeong-A Jang;Min-Sook Kang
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1095-1106
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    • 2023
  • This study investigated the anti-oxidative and anti-inflammatory effects of Aster glehni (AG) extract in RAW 264.7 cells and Caenorhabditis elegans. The total polyphenol and flavonoid contents were higher in the ethanol extracts than in the hot water extracts. As a result of measuring the moisture contents (%) and extraction yields (%) of AG and drying A. glehni for processing (DAG), 70% ethanol, which has the highest percentage of extraction yield, was selected as the final solvent. DPPH radical scavenging activity showed higher antioxidant activity of ethanol extracts of DAG than AG. The cytotoxicity assay of the AG or DAG ethanol extracts was treated at different concentrations (25, 50, and 100 ㎍/mL), and cell viability rates were higher than 80% at all concentrations. The LPS-stimulated nitric oxide (NO) production in RAW 264.7 was significantly reduced at all concentrations of AG and DAG groups. As a result of measuring the gene expression of iNOS, which induces NO production, the AG or DAG group decreased by 33% and 32%, compared with the phosphate buffer saline (PBS) group. Under inflammatory stress conditions, the survival rate of C. elegans treated with AG or DAG ethanol extract with LPS showed concentration-dependent improvement in survival rate compared with the PBS group. Considering these results, AG could potentially be developed as an antioxidant and anti-inflammatory functional food material.

Research on Immune Responses Induced by Salmonella Typhimurium Infectionin CRIP1-Deficient Condition (CRIP1결손조건 하에서 Salmonella Typhimurium 감염에 의해 유도되는 면역반응에 관한 연구)

  • Dongju Seo;Se-Hui Lee;Sun Park;Hyeyun Kim;Jin-Young Yang
    • Journal of Life Science
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    • v.34 no.1
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    • pp.48-58
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    • 2024
  • Salmonella is a common food-borne intracellular bacterial pathogen that has triggered significant public health concerns. Salmonella hosts' genetic factors play a pivotal role in determining their susceptibility to the pathogen. Cysteine-rich intestinal protein 1 (CRIP1), a member of LIM/double zinc finger protein family, is widely expressed in humans, such as in the lungs, spleen, and especially the gut. Recently, CRIP1 has been reported as a key marker of several immune disorders; however, the effect of CRIP1 on bacterial infection remains unknown. We aimed to elucidate the relationship between Salmonella infection and CRIP1 gene deficiency, as Salmonella spp. is known to invade the Peyer's patches of the small intestine, where CRIP1 is highly expressed. We found that CRIP1-deficient conditions could not alter the characteristics of bone marrow-derived myeloid cells in terms of phagocytosis on macrophages and the activation of costimulatory molecules on dendritic cells using ex vivo differentiation. Moreover, flow cytometry data showed comparable levels of MHCII+CD11b+CD11c+ dendritic cells and MHCII+F4/80+CD11b+ macrophages between WT and CRIP1 knockout (KO) mice. Interestingly, the basal population of monocytes in the spleen and neutrophils in MLNs is more abundant in a steady state of CRIP1 KO mice than WT mice. Here, we demonstrated that the CRIP1 genetic factor plays dispensable roles in host susceptibility to Salmonella Typhimurium infections and the activation of myeloid cells. In addition, differential immune cell populations without antigen exposure in CRIP1 KO mice suggest that the regulation of CRIP1 expression may be a novel immunotherapeutic approach to various infectious diseases.

Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level (전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.

Study on skin anti-inflammatory activity of fig (Ficus carica L.) fruit extract fractions (무화과(Ficus carica L.) 열매 추출 분획의 피부 항염증 활성 연구)

  • Hee Joon Kwon;Geun soo Lee;Jin Hwa Kim;Soon Woo Kwon;Hyung seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.416-423
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    • 2023
  • Figs has known to have antioxidant, whitening, anti-inflammatory, and antibacterial effects in their leaves, roots, stems, latex, and fruits. In order to develop cosmetic materials based on natural products, we have studied on the skin activity of the ficin in latex as well as the whitening function of the fruit extract with 70% ethanol, and used it as a raw material for released cosmetic product. However, there is little research on the demand for the development of new eutectic solvent extraction methods and its ability to control skin inflammation and psoriasis regulation. Thus, in this study, we evaluated the effectiveness of fig fruit extracts and fractions using eutectic solvent extraction for skin inflammation control and psoriasis. First, fig fruits were extracted under optimal eutectic solvent conditions and fractionated with n-hexane, dichloromethane, ethyl acetate, and butanol. First, the antioxidant activity and inhibition of nitric oxide (NO) production were confirmed in mouse macrophage RAW264.7 cells. In addition, as a result of observing the mRNA expression through RT-PCR, pro-inflammatory cytokines such as TNF-α, IL1α, and IL-1β were suppressed significantly in the hexane, dichloromethane, and ethyl acetate fractions. In addition, it was confirmed in TNF-α stimulated HaCaT keratinocyte model. Finally, chemokine CC motif ligand 20 (CCL20), marker gene of human psoriasis skin disease, was significantly suppressed in the hexane, dichloromethane, and ethyl acetate fractions. These results suggested its anti-inflammatory and skin soothing effect and the possibility of development as an excellent skin soothing natural cosmetic material in the future through future clinical trials.