• Journal of Korean Society of Forest Science
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• v.84 no.1
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• pp.77-86
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• 1995

To effectively modify tree function with genetic engineering, transgenes must be expressed at the proper level in the appropriate tissues at suitable developmental stages. Toward understanding the spatial and temporal expression of transgenes in woody plants, transgene expression was evaluated in three greenhouse-grown, transgenic lines of Populus alba ${\times}$ P. grandidentata hybrid clone 'Hansen'. All transgenic poplar lines possess constructs containing the bacterial nopaline synthase(nos) promoter linked to a neomycin phosphotransferase II(NPT II) selectable marker gene. In addition, each transgenic poplar line contains one of the following gene constructs : 1) a wound-inducible potato proteinase inhibitor II (pin2) promoter linked to a chloramphenicol acetyltransferase(CAT) reporter gene. 2) a nos promoter linked to a PIN2 structural gene : or 3) a Cauliflower Mosaic Virus 35s promoter linked to a PIN2 structural gene. Polymerase chain reaction(PCR) was used to verify the presence of foreign genes in the poplar genome. Enzyme-linked immunosorbent assays(ELISAs) were used to evaluate organ specific expression of the nos-NPT II construct. NPT II expression was detected in leaves, petioles, stems, and roots of transgenic poplar, thereby indicating that the nos promoter is potentially effective for general constitutive expression of transgenes. NPT expression varied among transgenic poplar lines and among organs for one transgenic line, Tr15. With Tr15, NPT II levels were highest in older leaves and petioles. These results indicate that screening of several transgenic lines may be required to identify lines with optimal transgene expression.

• Animal cells and systems
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• v.2 no.1
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• pp.117-122
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• 1998

D-raf, a Drosophila homolog of the human c-raf-1, is known as a signal transducer in cell proliferation and differentiation. A previous study found that the D-raf gene expression is regulated by the DNA replication-related element (DRE)/DRE-binding factor (DREF) system. In this study, we found the sequences homologous to transcription factor C/EBP, MyoD, STAT and Myc recognition sites in the D-raf promoter. We have generated various base substitutional mutations in these recognition sites and subsequently examined their effects on D-raf promoter activity through transient CAT assays in Kc cells with reporter plasmids p5'-878DrafCAT carrying the mutations in these binding sites. Through gel mobility shift assay using nuclear extracts of Kc cells, we detected factors binding to these recognition sites. Our results show that transcription factor C/EBP, STAT and Myc binding sites in D-raf promoter region play a positive role in transcriptional regulation of the D-raf gene and the Myo D binding site plays a negative role.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.271-276
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• 2008

Ribosome stalling by nascent sticky peptide has been reported in several organisms across the kingdom. To test whether small subunit (SSU) rRNA is involved in this phenomenon, we developed a genetic system that utilized the specialized ribosome system to isolate SSU rRNA mutants that enable ribosomes to bypass the SecM-derived sticky peptide in protein synthesis. In this system, CAT-SecM mRNA, which encodes CAT protein containing the sticky peptide derived from SecM, is only translated by specialized ribosomes. These ribosomes were shown to transiently stall on CAT-SecM mRNA followed by the synthesis of the sticky peptide. Expression of specialized ribosomes resulted in the decreased steady-state level of CAT-SecM mRNA, which is consistent with a notion that ribosome stalling induces mRNA degradation. Isolation and characterization of SSU rRNA mutations using this genetic system that are sufficient to circumvent ribosome stalling induced by the SecM-derived sticky peptide will provide evidence of SSU rRNA function in mRNA cleavage.

• Journal of Veterinary Clinics
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• v.37 no.4
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• pp.208-212
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• 2020

A 6-year-old castrated male Russian Blue cat was presented for evaluation of dyschezia. Abdominal ultrasound revealed hyperechoic nodules in both kidneys, heterogeneous mass in abdomen, and extensive mesenteric thickening with multiple hypoechoic nodules. Computed tomography showed multiple hypodense lesions in both kidneys and diffuse nodular infiltration around the mesentery. Fine needle aspirates (FNA) acquired under ultrasound guidance from the mesentery consisted of large lymphocytes which have round to irregular nuclei with granular chromatin, prominent nucleoli and a small amount of basophilic cytoplasm. Polymerase chain reaction (PCR) for antigen receptor gene rearrangement result of FNA sample revealed a T-cell malignancy. The cat died from acute renal failure after 1 cycle of modified Madison-Wisconsin L-CHOP protocol. Postmortem examination revealed bilaterally enlarged lumpy-bumpy shaped kidneys. Histopathologic examination showed an infiltration of malignant lymphocytes into the renal parenchyma and mesentery. Immunohistochemical staining of the renal sample displayed a negative expression of CD3, PAX5, MUM-1, and CD79. The clinical features and prognosis of the cat with renal lymphoma with mesenteric lymphomatosis have been described in this report.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.5
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• pp.520-529
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• 2014

The purpose of this study is to investigate the protective effects of Yukmijiwhang-tang(YM) water extracts against the UVB irradiation on the human keratinocyte HaCaT cells. We observed the effects of YM on the oxidative stress, gene expression of pro-inflammatory cytokine such as TNF-${\alpha}$ and IL-$1{\beta}$, and matrix metalloproteinase-9 in UVB-irradiated HaCaT cells. On the effects of oxidative stress and antioxidant function on the treatment with YM, The activity of xanthine oxidase(XO) was significantly decreased by treatment of YM in all the concentrations(p<0.01). The activity of superoxide dismutase(SOD) and catalase(CAT) was significantly increased by treatment of YM in a dose dependent manner(p<0.05 and p<0.01). DPPH radical was erased by treatment of YM under dose of $500{\mu}g/m{\ell}$ concentration. Treatment of HaCaT cells with YM had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(p<0.05, p<0.01, p<0.001). Gelatin zymography assay showed that YM downregulated the MMP-9 activity in UVB-irradiated HaCaT cells. RT-PCR analysis revealed that YM suppressed the expression of IL-$1{\beta}$ and MMP-9 however, it has no effects on the expression of TNF-${\alpha}$ and MMP-3. Our study suggests that Yukmijiwhang-tang exert protective actions on the UVB-irradiated HaCaT cells largely by anti-oxidative and anti-inflammatory processes.

• Journal of Haehwa Medicine
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• v.23 no.2
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• pp.5-13
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• 2015

Objectives : The anti-inflammatory and antioxidant effects of water extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) were investigated. The effects of SSJ and GSJ were compared. Methods : We performed cell viability assay in HaCaT cells and RAW 264.7 cells using 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide(MTT) assay. We measured chemokines(regulated on activation normal T-cell expression and secreted ; RANTES/CCL5, interferon-inducible protein; IP-10/CXCL10, macrophage-derived chemokine; MDC/CCL22) in HaCat cells, also we measured cytokines (tumor necrosis factor-${\alpha}$; TNF-${\alpha}$, interleukin-6; IL-6) and nitric oxide(NO) production in RAW 264.7 cells using enzyme-linked immunosorbent assay(ELISA) and NO assay. Western blot assay was used to evaluate the expression for inducible nitric oxide synthase(iNOS) in RAW 264.7 cells. Results : SSJ and GSJ did not affect the cell viability at the concentrations treated ($0-800{\mu}g/ml$). As a result of SSJ and GSJ treatment in HaCat cells stimulated by TNF-${\alpha}$(10 ng/ml) and interferon(IFN)-${\gamma}$(10 ng/ml), the production of RANTES and IP-10 was inhibited significantly. However there was no significant difference in the secretion of MDC. And in RAW 264. 7 cells stimulated by lipopolysaccharide(LPS, $1{\mu}g/ml$), SSJ and GSJ treatment significantly inhibited the secretion of TNF-${\alpha}$ and IL-6 and the production of NO. The expression of iNOS was also decresed by SSJ and GSJ treatment in RAW 264. 7 cells. Compared with SSJ, GSJ was superior to SSJ in inhibition of RANTES, IP-10, TNF-${\alpha}$, IL-6 and NO production at the concentration of $200{\mu}g/ml$. Conclusion : Both SSJ and GSJ have anti-inflammatory and antioxidant effects. And GSJ has better effects than SSJ.

• Journal of Animal Science and Technology
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• v.58 no.11
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• pp.39.1-39.12
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• 2016

Background: The current study investigates the anti-stress effects of clove (Eugenia caryophyllus) extracts (0, 200, 400, and 600 mg/kg) on serum antioxidant biomarkers, immune response, immunological organ growth index, and expression levels of acute phase proteins (APPs); ovotransferrin (OVT), ceruloplasmin (CP), ceruloplasmin (AGP), C-reactive protein (CRP), and serum amyloid-A (SAA) mRNA in the immunological organs of 63-d-old male black-meated Silkie fowls subjected to 21 d chronic heat stress at $35{\pm}2^{\circ}C$. Results: The results demonstrated that clove extract supplementation in the diet of Silkie fowls subjected to elevated temperature (ET) improve growth performance, immune responses, and suppressed the activities of glutathion peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and thioredoxin reductase (TXNRD); reduced serum malonaldehyde (MDA) and glutathione (GSH) concentrations when compared with fowls raised under thermoneutral condition (TC). Upon chronic heat stress and supplementation of clove extracts, the Silkie fowls showed a linear increase in GSH-Px, SOD, CAT, and TXNRD activities (P = 0.01) compared with fowls fed diets without clove extract. ET decreased (P < 0.05) the growth index of the liver, spleen, bursa of Fabricius and thymus. However, the growth index of the liver, spleen, bursa of Fabricius and thymus increased significantly (P < 0.05) which corresponded to an increase in clove supplemented levels. The expression of OVT, CP, AGP, CRP, and SAA mRNA in the liver, spleen, bursa of Fabricius and thymus were elevated (P < 0.01) by ET compared with those maintained at TC. Nevertheless, clove mitigates heat stress-induced overexpression of OVT, CP, AGP, CRP and SAA mRNA in the immune organs of fowls fed 400 mg clove/kg compared to other groups. Conclusions: The results showed that clove extracts supplementation decreased oxidative stress in the heat-stressed black-meated fowls by alleviating negative effects of heat stress via improvement in growth performance, antioxidant defense mechanisms, immunity, and regulate the expression of acute phase genes in the liver and immunological organs.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.4
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• pp.663-668
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• 2011

Galgeun-tang (GGT) has been a great source for treating cold diseases in the folk medicine recipe. Carbon tetrachloride ($CCl_4$) is one type of hepatotoxin that can eventually cause liver injury. During the experiment, we first studied the protective effects of GGT against $CC_4$-induced hepatotoxicity. GGT was pretreated for 3 h, and 1% $CCl_4$ was added to mouse primary liver cells. After 4 h, ROS generation and expression of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)) were analyzed by FACS and real time PCR. Also, the activities of ALT and LDH were measured using cultured medium. The hepatic levels of TNF-alpha and iNOS, which are related to inflammation and stress response gene, HSP72 and HO-1 were analyzed by PCR or real time PCR. Liver tissues were analyzed by HE stain. From the observation, we discovered that GGT treatment protects $CCl_4$-induced hepatotoxicity, and that GGT pretreatment decreases ROS generation, TNF-alpha and iNOS expression. However, gene expression of CAT, SOD, GPx, HSP72 and HO-1 were increased by GGT. These results lead to the conclusion that GGT has protective effects against $CCl_4$-induced hepatotoxicity.

• Animal Bioscience
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• v.34 no.11
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• pp.1766-1775
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• 2021

Objective: The oxidative stress status and changes of chicken ovary tissue after shading were studied, to determine the mechanism of the effect of shading on follicular development. Methods: Twenty healthy laying hens (40 weeks old) with uniform body weight and the same laying rate were randomly divided into two groups (the shading group and normal light group). In the shading group, the cage was covered to reduce the light intensity inside the cage to 0 without affecting ventilation or food intake. The normal lighting group received no additional treatment. After 7 days of shading, oxidative stress related indicators and gene expression were detected. Results: Analysis of paraffin and ultrathin sections showed that apoptosis of ovarian granulosa cells (GCs) increased significantly after light shading. Enzyme linked immunosorbent assay results revealed that the levels of total antioxidant capacity, malondialdehyde, superoxide dismutase (SOD), glutathione, catalase (CAT), and other substances in the sera, livers, ovaries, and follicular GCs of laying hens increased significantly after shading for 7 days; and reactive oxygen species (ROS) levels in the livers of laying hens also increased significantly. ROS in the serum, ovarian and GCs also increased. After shading for 7 days, the levels of 8-hydroxy-2 deoxyguanosine in the sera and ovarian tissues of laying hens increased significantly. Cell counting kit-8 detection showed that the proliferation activity of GCs in layer follicles decreased after shading for 7 days; the expression level of the anti-apoptotic gene B-cell lymphoma-2 in ovarian tissue and follicular GCs was significantly reduced, and the expression levels of pro-apoptotic caspase 3 (casp3), and SOD, glutathione peroxidase 2 (GPX2), and CAT were all significantly increased. Conclusion: Oxidative stress induced by shading light has a serious inhibitory effect on follicular development during reproduction in laying hens.

• Korean Journal of Acupuncture
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• v.38 no.1
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• pp.32-42
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• 2021

Objectives : In this study, we investigated the neuroprotective effects of ethanol extract of Lonicera japonica flower buds (EELJ) on glutamate-induced neurotoxicity in mouse hippocampus-derived neuronal HT22 cells. Methods : After analyzing the cytoprotective effect of EELJ on glutamate in HT22 cells, the inhibitory effect of apoptosis was studied using flow cytometry. In order to analyze the antioxidant efficacy of EELJ, the levels of reactive oxygen species (ROS) and glutathione (GSH) were investigated, and the effects on the activities of superoxide dismutase (SOD) and catalase (CAT) were also analyzed. Furthermore, the effect of EELJ on the expression of apoptosis regulators such as Bax and Bcl-2 in glutamate-treated HT22 cells was investigated. Results : According the current results, pretreatment with EELJ significantly reduced glutamate-induced loss of cell viability and release of lactate dehydrogenase. EELJ also markedly attenuated glutamate-induced generation of intracellular ROS, which was associated with increased levels of GSH, and activity of SOD and CAT in glutamate-stimulated HT22 cells. In addition, EELJ was strikingly inhibited glutamate-induced apoptosis in HT22 cells. Furthermore, the expression of pro-apoptotic Bax was increased and the expression of anti-apoptotic Bcl-2 was decreased in glutamate-treated HT22 cells, while in the presence of EELJ, their expressions were maintained at the control levels. Conclusions : These findings indicate that EELJ protects glutamate-induced cytotoxicity in HT22 hippocampal neurons through antioxidant activity. Therefore, although identification of biologically active substances of EELJ and re-evaluation through animal experiments is necessary, this natural substance is a promising candidate for further research in preventing and treating oxidative stress-mediated neurodegenerative diseases.