• Title/Summary/Keyword: DNA protection assay

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Antioxidative Activity and Protection of Oxidative Chromosomal Damage by Vegetables, Fruits Extract and Their Functional Liquid Formulation (야채 및 과일추출물의 항산화작용과 산화적 염색체손상에 대한 억제효과)

  • 이승철;허찬;이승현;김현표;허문영
    • YAKHAK HOEJI
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    • v.48 no.2
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    • pp.111-116
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    • 2004
  • The ethanol extracts of mixed vegetables (Bioactive Vegetables, BV), mixed fruits (Bioactive Fruits, BF) and their liquid formulation (Chungpae Plus , CP) were evaluated for their antioxidative and antigenotoxic activities. They were shown to possess the significant free radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and were revealed to show the inhibitory effect of lipid peroxidation as measured by malondialdehyde (MDA) formation although the potencies were not higher than those of vitamin C. They did not possess any pro-oxidant effect on bleomycin-Fe(III) dependent DNA degradation, whereas vitamin C showed strong pro-oxidant effect. Furthermore, oral administration of BV and BF inhibited micronucleated reticulocyte (MNRET) formation of mouse peripheral blood induced by KBrO3 treatment in vivo. CP also showed significant inhibition under same experimental condition. Therefore, the liquid formulation (CP) containing BV and BF may be a useful natural antioxidative and antigenotoxic agent by scavenging free radicals, inhibition of lipid peroxidation and protecting chromosomal damage.

Replication and encapsidation of recombinant Turnip yellow mosaic virus RNA

  • Shin, Hyun-Il;Kim, In-Cheol;Cho, Tae-Ju
    • BMB Reports
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    • v.41 no.10
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    • pp.739-744
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    • 2008
  • Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus that infects mainly Cruciferae plants. In this study, the TYMV genome was modified by inserting an extra subgenomic RNA promoter and a multiple cloning site. This modified TYMV was introduced into Nicotiana benthamiana using a Agrobacterium-mediated T-DNA transfer system (agroinfiltration). When a gene encoding $\beta$-glucuronidase or green fluorescent protein was expressed using this modified TYMV as a vector, replication of the recombinant viruses, especially the virus containing $\beta$-glucuronidase gene, was severely inhibited. The suppression of replication was reduced by co-expression of viral silencing suppressor genes, such as tombusviral p19, closteroviral p21 or potyviral HC-Pro. As expected, two subgenomic RNAs were produced from the recombinant TYMV, where the larger one contained the foreign gene. An RNase protection assay revealed that the recombinant subgenomic RNA was encapsidated as efficiently as the genuine subgenomic RNA.

Synthesis and Characterization of Degradable Polycationic Polymers as Gene Delivery Carriers

  • Kim, Hyun-Jin;Kwon, Min-Sung;Choi, Joon-Sig;Kim, Bo-Hye;Yoon, Jae-Keun;Kim, Kwan;Park, Jong-Sang
    • Bulletin of the Korean Chemical Society
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    • v.28 no.1
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    • pp.63-67
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    • 2007
  • Biodegradable cationic poly(ester-amide) polymers were synthesized by double-monomer method, that showed excellent solubility in many organic solvents and water. Different degradation patterns were obtained by the regulation of monomer ratios and overall long period of time of DNA protection up to 12 days was shown by PicoGreen reagent assay. Good transfection profiles in the presence of serum and very low toxicity on mammalian cells may allow these polymers to become suitable for long-term gene delivery systems and therapeutic applications.

Wisteria Vein Mosaic Virus Detected for the First Time in Iran from an Unknown Host by Analysis of Aphid Vectors

  • Valouzi, Hajar;Hashemi, Seyedeh-Shahrzad;Wylie, Stephen J.;Ahadiyat, Ali;Golnaraghi, Alireza
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.87-97
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    • 2020
  • The development of reverse transcription-polymerase chain reaction using degenerate primers against conserved regions of most potyviral genomes enabled sampling of the potyvirome. However, these assays usually involve sampling potential host plants, but identifying infected plants when they are asymptomatic is challenging, and many plants, especially wild ones, contain inhibitors to DNA amplification. We used an alternative approach which utilized aphid vectors and indicator plants to identify potyviruses capable of infecting common bean (Phaseolus vulgaris). Aphids were collected from a range of asymptomatic leguminous weeds and trees in Iran, and transferred to bean seedlings under controlled conditions. Bean plants were tested serologically for potyvirus infections four-weeks postinoculation. The serological assay and symptomatology together indicated the presence of one potyvirus, and symptomology alone implied the presence of an unidentified virus. The partial genome of the potyvirus, encompassing the complete coat protein gene, was amplified using generic potyvirus primers. Sequence analysis of the amplicon confirmed the presence of an isolate of Wisteria vein mosaic virus (WVMV), a virus species not previously identified from Western Asia. Phylogenetic analyses of available WVMV sequences categorized them into five groups: East Asian-1 to 3, North American and World. The Iranian isolate clustered with those in the World group. Multiple sequence alignment indicated the presence of some genogroup-specific amino acid substitutions among the isolates studied. Chinese isolates were sister groups of other isolates and showed higher nucleotide distances as compared with the others, suggesting a possible Eastern-Asian origin of WVMV, the main region where Wisteria might have originated.

Phylogeny, Morphology and Pathogenicity of Biscogniauxia mediterranea Causing Charcoal Canker Disease on Quercus brantii in Southern Iran

  • Samaneh, Ahmadi;Fariba, Ghaderi;Habiballah, Charehgani;Soraya, Karami;Dariush, Safaee
    • Research in Plant Disease
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    • v.28 no.4
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    • pp.209-220
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    • 2022
  • Charcoal canker of oak, which has recently increased in southern Iran, could pose a serious threat to the entire forest ecosystem in the near future. In addition, it seems that climate change and its consequences, such as drought in the southern regions of Iran, have exacerbated this phenomenon. Consequently, the objective of this study was to identify the fungal pathogens that could cause charcoal canker disease in the oak forests of South Zagros. It was also sought to find associations between changes in the occurrence/exacerbation of charcoal canker disease under non and intense drought stress in non-inoculated or inoculated Quercus brantii seedlings. In total, 120 isolates were obtained from eight oak forests located in the Zagros Mountains of Southern Iran, Kohgiluyeh & Boyer-Ahmad and Fars provinces, which were classified as Biscogniauxia mediterranea based on morphological assessment. Subsequently, molecular assay confirmed the result by phylogenetic inference of internal transcribed spacer-rDNA regions, α-actin, and β-tubulin genes. The results of the pathogenicity test showed that the response of isolates of B. mediterranea (Iran-G1 and Iran-M70) was varied in different environments for the measured necrotic lesion length. In comparison with the control moisture treatments (non-stress), the necrotic lesion length in inoculated treatments increased under intense drought stress. In general, inoculated oak seedlings' exposure to water-deficient stress by the pathogen of B. mediterranea could affect the spread/severity of the charcoal canker disease.

Differentially Expressed Genes by Methylmercury in Neuroblastoma cell line using suppression subtractive hybridization (SSH) and cDNA Microarray

  • Kim, Youn-Jung;Chang, Suk-Tai;Yun, Hye-Jung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.187-187
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    • 2003
  • Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, two methods, cDNA Microarray and SSH, were performed to assess the expression profile against MeHg and to identify differentially expressed genes by MeHg in neuroblastoma cell line. TwinChip Human-8K (Digital Genomics) was used with total RNA from SH-SY5Y (human neuroblastoma cell line) treated with solvent (DMSO) and 6.25 uM (IC50) MeHg. And we performed forward and reverse SSH method on mRNA derived from SH-SY5Y treated with DMSO and MeHg (6.25 uM). Differentially expressed cDNA clones were sequenced and were screened by dot blot and ribonuclease protection assay to confirm that individual clones indeed represent differentially expressed genes. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as environmental pollutants.

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Molecular Cloning and Identification of Novel Genes, Gomsin, Characteristically Expressed in Snailfish, Liparis tanakae (꼼치에서 특징적으로 발현되는 새로운 유전자 곰신의 분리 및 동정)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.6 no.1
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    • pp.7-16
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    • 2002
  • In order to obtain the specific genes of snailfish a subtracted cDNA library was constructed, and analysed by sequencing and GenBank search. Among them C90-171 clone was turned out to be genes showing low homology and nonredundant genes. This novel clone was named Gomsin(C90-171). Gomsin was shown to be intensely expressed in the epithelial cells, some mesenchymal cells, and sheaths of muscle bundles in the result of immunohistochemistry. In the cross reaction assay of Gomsin antibody against various human tissues, the Gomsin was strongly expressed in the ductal and acinar cells of salivary glands, which was similar to the expression patterns of proline-rich proteins(PRPs) of human. The antibody raised against the Gomsin was clearly cross-reacted with human salivary PRPs and also recombinant proteins of human PRPs in the Western blot and immunoprecipitation analysis. Contrast to the salivary PRPs, the Gomsin was not easily degraded in the mixed saliva, but rapidly attacked on the cultured keratocytes in vitro. The simulated protein structure of Gomsin was similar to the whorled pattern of PRPs, even though the amino acid sequence of Gomsin was quite different from those of PRPs. These data suggest that the Gomsin is a characteristic matrix protein in the skin and body of snailfish, which is also utilized for the tissue protection in the similar way to the PRPs of human muco-secretory organs.

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Biological Activities of Flavonoid Glycosides Isolated from Angelica keiskei (신선초에서 분리된 flavonoid glycosides의 생리활성)

  • Shim, Jae-Seok;Kim, Seung-Deok;Kim, Tae-Seok;Kim, Kyung-Nam
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.78-83
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    • 2005
  • Recently, much attention has been focused on plant antioxidants, because they are expected to protect against oxidative damage, possibly preserving biological functions of cells. Antioxidant compounds were isolated from Angelica keiskei through extraction with 80% EtOH, and fractionations were carried out sequentially with n-hexane, chloroform, ethyl acetate, n-butanol, and water. Two active compounds were isolated from ethyl acetate fraction by silica gel column chromatography, and were identified as isoquercitrin ($quercetin-3-O-{\beta}-D-glucose$) and hyperoside ($quercetin-3-O-{\beta}-D-glucose$). Isoquercitrin and hyperoside showed strong antioxidative potency, as revealed by evaluation of their ABTS, DPPH, OH, and $H_{2}O_{2}$ radical-scavenging activities, and ex vivo DNA damage-protecting effects.

Natural Occurrence of Tomato leaf curl New Delhi virus in Iranian Cucurbit Crops

  • Yazdani-Khameneh, Sara;Aboutorabi, Samaneh;Shoori, Majid;Aghazadeh, Azin;Jahanshahi, Parastoo;Golnaraghi, Alireza;Maleki, Mojdeh
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.201-208
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    • 2016
  • The main areas for field-grown vegetable production in Iran were surveyed during the years of 2012-2014 to determine the occurrence of begomoviruses infecting these crops. A total of 787 leaf samples were collected from vegetables and some other host plants showing virus-like symptoms and tested by an enzymelinked immunosorbent assay (ELISA) using polyclonal antibodies produced against Tomato yellow leaf curl virus (TYLCV). According to the ELISA results, 81 samples (10.3%) positively reacted with the virus antibodies. Begomovirus infections were confirmed by polymerase chain reaction (PCR) using previously described TYLCV-specific primer pair TYLCV-Sar/TYLCV-Isr or universal primer pair Begomo-F/Begomo-R. The PCR tests using the primer pair TYLCV-Sar/TYLCV-Isr resulted in the amplification of the expected fragments of ca. 0.67-kb in size for ELISA-positive samples tested from alfalfa, pepper, spinach and tomato plants, confirming the presence of TYLCV. For one melon sample, having a week reaction in ELISA and no reaction in PCR using TYLCV-specific primers, the PCR reaction using the primer pair Begomo-F/Begomo-R resulted in the amplification fragments of the expected size of ca. 2.8 kb. The nucleotide sequences of the DNA amplicons derived from the isolate, Kz-Me198, were determined and compared with other sequences available in GenBank. BLASTN analysis confirmed the begomovirus infection of the sample and showed 99% identities with Tomato leaf curl New Delhi virus (ToLCNDV); phylogenetic analysis supported the results of the database searches. This study reports the natural occurrence of TYLCV in different hosts in Iran. Our results also reveal the emergence of ToLCNDV in Iranian cucurbit crops.

Study on Antitumor Activity and Radioprotective effects of Kamisasammaekmundongtang (가미사삼맥문동탕(加味沙蔘麥門冬湯)이 항암활성(抗癌活性)과 방사선부작용(放射線副作用) 억제효과(抑制效果)에 미치는 영향(影響))

  • Park, Yang-chun;Kim, Byeong-tak
    • Journal of Haehwa Medicine
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    • v.8 no.1
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    • pp.403-424
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    • 1999
  • To evaluate the antitumor activity, antimetastatic and radioprotective effects of Kamisasammaekmundongtang(KSMT), studies were done experimentally. The results were obtained as follows: 1. In cytotoxicity against P388, A549 and B16-F10, KSMT was not showed satisfiable cytotoxicity as compared with control. 2. In Inhibitory effect on activity of DNA topoisomerase I, KSMT has strong inhibitory effect. 3. The inhibitory effect on adhesion of A549 to complex extracellular matrix was significantly increased at 0.5mg/ml, 1mg/ml of KSMT. 4. The T/C% was 122 in KSMT treated group in S-180 bearing ICR mice. 5. In antiangiogenetic effect on CAM assay, inhibitory rate was 33% in KSMT treated group. 6. In pulmonary colonization assay, a number of colonies in the lungs were decreased significantly in KSMT treated group as compared with control group. 7. By FACS analysis of splenic leukocyte after exposure to radiation by linear accelerator, T-helper cell, B cell and macrophage in KSMT treated group were significantly increased while splenocytes were decreased in control group. 8. In histological changes of jejunum of $Bald{\setminus}C$ mice after exposure to radiation by linear accelerator, exclusion and fusion of villi were decreased as compared with control group. But in duodenum and ileum, exclusion and fusion of villi were not decreased as compared with control group. 9. WBC, PLT were increased in KSMT treated group as compared with control group after exposure to radiation by linear accelerator, but the increasing effect was not significant. Above results suggest that KSMT may be useful in prevention of cancer metastasis and protection from damage by radiotherapy. But the further study of KSMT would be demanded.

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