• Title/Summary/Keyword: DNA in tail

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Protective and Anti-Pathology Effects of Sm Fructose-1,6-Bisphosphate Aldolase-Based DNA Vaccine against Schistosoma mansoni by Changing Route of Injection

  • Saber, Mohamed;Diab, Tarek;Hammam, Olft;Karim, Amr;Medhat, Amina;Khela, Mamdouh;El-Dabaa, Ehab
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.155-163
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    • 2013
  • This study aimed to evaluate the efficacy of fructose-1,6-bis phosphate aldolase (SMALDO) DNA vaccination against Schistosoma mansoni infection using different routes of injection. The SMALDO has been cloned into the eukaryotic expression vector pcDNA3.1/V5-His TOPO-TA and was used in injecting Swiss albino mice intramuscularly (IM), subcutaneously (SC), or intraperitoneally (IP) ($50{\mu}g/mouse$). Mice vaccinated with non-recombinant pcDNA3.1 served as controls. Each group was immunized 4 times at weeks 0, 2, 4, and 6. Two weeks after the last booster dose, all mice groups were infected with 80 S. mansoni cercariae via tail immersion. At week 8 post-infection, animals were sacrificed for assessment of parasitological and histopathological parameters. High anti-SMALDO IgG antibody titers were detected in sera of all vaccinated groups (P<0.01) compared to the control group. Both the IP and SC vaccination routes resulted in a significant reduction in worm burden (46.2% and 28.9%, respectively, P<0.01). This was accompanied by a significant reduction in hepatic and intestinal egg counts (41.7% and 40.2%, respectively, P<0.01) in the IP group only. The number of dead eggs was significantly increased in both IP and IM groups (P<0.01). IP vaccination recorded the highest significant reduction in granuloma number and diameter (54.7% and 29.2%, respectively, P<0.01) and significant increase in dead miracidia (P<0.01). In conclusion, changing the injection route of SMALDO DNA vaccination significantly influenced the efficacy of vaccination. SMALDO DNA vaccination via IP route could be a promising protective and antipathology vaccine candidate against S. mansoni infection.

Comet Assay to Detect the DNA Breakages in the Tissue of the Purple Clam ( Saxidomus purpuratus) and the Blood of the Olive Flounder (Paralichthys olivaceus) Exposed to 5 PAHs

  • Lee, Taek-Kyun;Kim, So-Jung;Park, Eun-Seok;Rora Oh;Yun, Hee-Young;Man Chang
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.159-159
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    • 2003
  • Comet assay is a potential monitoring tool because DNA strand breakage may be produced by a wide range of agents. The comet assay, also called the single-cell gell electrophoresis (SCGE) assay, is rapid and sensitive method for the detection of DNA damage in cells. This study was performed for the identification of DNA damage in the cells from flounders and clams exposed to PAHs. As a control experiments, flounder and clam cells were exposed to $H_2O$$_2$. The cells exposed to $H_2O$$_2$ were displayed a typical nuclei movement DNA damage of cells were significantly increased when the isolated cells from the blood of flounders and the tissue of clams were in vitro exposed to the different concentrations (5, 10, 50, 100 ppb) of five kinds of PAHs (benzo[a]pyrene, pyrene, fluoranthene, anthrancene, and phenanthrene). For the in vivo test, flounders and clams were exposed to the different concentrations of BaP for 4 days. The results showed that DNA strand breakage was effected by the concentration of BaP and the duration of exposure. In high concentration of BaP, the mean tail lengths of nuclei was longer than it In low concentration, while the mean size of head DNA decreased. In this research, both in vitro and in vivo genotoxicity of PAHs could be biomonitored by the comet assay. Especially, clams and flounders seem to be useful as materials for monitoring genotoxic damage by comet assay.

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Effects of Gamma-ray Irradiation on Growth Characteristics and DNA Damage in Licorice (Glycyrrhiza uralensis) (감마선 조사가 감초(Glycyrrhiza uralensis)의 초기 생육 및 DNA 손상에 미치는 영향)

  • Ryu, Jaihyunk;Im, Seung Bin;Kim, Dong Sub;Ahn, Joon-Woo;Kim, Jin-Baek;Kim, Sang Hoon;Kang, Si-Yong
    • Journal of Radiation Industry
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    • v.8 no.2
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    • pp.89-95
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    • 2014
  • This study was conducted to determine the optimal dose of gamma-ray on the growth and nucleus DNA damage for mutation breeding in licorice. Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy). Significant decreases in germination rate (%), survival rate (%) and growth characteristics (plant height, number of leaves, root length and fresh weight) were observed by dose of increased. $LD_{50}$ (lethal dose) was approximately 400 Gy to 500 Gy. Also, reduction doses ($RD_{50}$) of plant height, number of leaves, root length and flash weight were 428 Gy, 760 Gy, 363 Gy and 334 Gy, respectively. It is supplest that the optimal dose of gamma irradiation for licorice mutation induction might be about 400 Gy in this study. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 300 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 1000 Gy, the rate of head DNA was decreased significantly from 92.88% to 73.09%. Tail length(${\mu}m$) was increased as the dose of increased over 300 Gy. Growth characteristics (Germination rate, Survival rate, plant height, number of leaves, root length and fresh weight) were highly negatively ($P{\leq}0.01$) correlated with dose. While the tail length was highly positively ($P{\leq}0.01$) correlated with dose.

Establishment of the High-Throughput Hair Roots' DNA Isolation System and Verification of Its Appicability for Hanwoo Traceability Using the 11 Microsatellite Makes (대량 모근 시료 DNA 분리 체계 확립과 11 microsatellite maker를 사용하는 한우 생산이력제로의 적용가능성 검증)

  • Lim, Hyun-Tae;Lee, Sang-Ho;Yoo, Chae-Kyoung;Sun, Du-Won;Cho, In-Cheol;Yoon, Du-Hak;Yang, Dae-Young;Cheong, Il-Cheong;Lee, Jung-Gyu;Jeon, Jin-Tae
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.91-99
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    • 2010
  • We used a multiplex PCR primer set composed of 11 microsatellite (MS) markers and two sexing markers for gender detection. Genomic DNA extracted from hair roots of 3,510 Hanwoo were genotyped. Based on the 11MS markers, no animals had identical genotypes(TGLA227, BM2113, TGLA53, ETF10, SPS115, TGLA122, ETH3, ETH225, BM1824 and INRA23). The expected probability of identity among genotypes of random individuals (PI), the probability of identity among genotypes from random half-sibs ($PI_{half-sibs}$) and among genotypes of random individuals, and the probability of identity among genotypes from random sibs ($PI_{sibs}$) were estimated as $1.31{\times}10^{-23}$, $2.52{\times}10^{-16}$and $1.09{\times}10^{-6}$, respectively using the API-CALC program, version 1.0. We successfully completed the genotype analysis of 3,510 Hanwoo with a 3.93% genotyping failure rate. It was revealed that extracting DNA from the hair root was a time-efficient and cost-effective method to collect specimens for DNA isolation from live animals. This method also minimized stress for the animals during specimen collection. Among the hair roots from the back, belly, upper tail and lower tail, 5~13 hair roots of the lower tail led to the best genotype analysis results. Finally, we established a 96-well-format method of DNA preparation applicable for high- throughput genotype analysis.

Morphological Description and Molecular Identification of Juvenile Peristedion liorhynchus (Peristediidae, Pisces) from Jeju-do Island, Korea (한국 제주도 근해에서 출현한 황성대과(Peristediidae) 어류 1미기록종, Peristedion liorhynchus 치어의 형태기재 및 분자동정)

  • Jang, Seo-Ha;Kim, Jin-Koo;Yu, Hyo-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.794-799
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    • 2020
  • In April 2019, a single Peristediidae juvenile (14.1 mm SL) was collected from the waters off of Jeju-do Island, Korea. DNA barcoding identified the juvenile as Peristedion liorhynchus, an unrecorded species in Korea. P. liorhynchus has eight dorsal fin spines, 22 dorsal and 19 anal fin rays, and a long third pectoral fin ray that passes through the middle of the tail at the juvenile stage. Juvenile also have large heads, extensive head spination, and serrated edges on the ocular and parietal spines. This is the first record of P. liorhynchus in Korea; therefore, we propose the new Korean name, "Nam-bang-hwang-seong-dae".

Influence of Mercury on the Repair of Ionizing Radiation-induced DNA Damage in Coelomocytes of Eisenia fetida (이온화 방사선에 의해 손상된 Eisenia fetida 체강세포의 DNA 수복에 수은이 미치는 영향)

  • Ryu, Tae-Ho;Nili, Mohammad;An, Kwang-Guk;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.3
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    • pp.236-240
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    • 2011
  • Mercury known as quicksilver, is the most common cause of heavy metal toxicity. Toxicity caused by excessive mercury exposure is now being recognized as a widespread environmental problem and is continuing to attract a great deal of public concerns. The mercury genotoxicity could be its effect on DNA repair mechanisms, which constitute the defense system designated to protect genome integrity. The objective of this study is to confirm that mercuric chloride inhibits the repair of gamma ray-induced DNA damage. The earthworm of Eisenia fetida was chosen for this study because it is an internationally accepted model species for toxicity testing with a cosmopolitan distribution. Experiments were done to identify the levels of DNA damage and the repair kinetics in the coelomocytes of E. fetida irradiated with 20 Gy gamma rays alone or with gamma rays after 40 mg $kg^{-1}$ $HgCl_2$ treatment by means of the single cell gel electrophoresis assay. The Olive tail moments were measured during 0~96 hours after irradiation. The repair time in the animals treated with the combination of $HgCl_2$ and ionizing radiation was nearly five times longer than that in the animals treated with ionizing radiation alone. Also, E. fetida exposed to mercury showed a statistically lower repair efficiency of gamma ray-induced DNA damage. The results suggest that the mercury could even have deleterious effects on the DNA repair system. Influence of mercury on the DNA repair mechanisms has been confirmed by this study.

Use of the In Vivo Single-cell Gel Electrophoresis Assay for Evaluating Genotoxicity in Clam (Single-cell Gel Electrophoresis Assay에 의한 대합에서의 In Vivo 유전독성 평가)

  • Kim Il-Yang;Hyun Chang-Kee
    • Toxicological Research
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    • v.20 no.3
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    • pp.225-232
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    • 2004
  • The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01~1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.

Does conventional freezing affect sperm DNA fragmentation?

  • Le, Minh Tam;Nguyen, Thai Thanh Thi;Nguyen, Tung Thanh;Nguyen, Trung Van;Nguyen, Tam An Thi;Nguyen, Quoc Huy Vu;Cao, Thanh Ngoc
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.2
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    • pp.67-75
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    • 2019
  • Objective: Sperm cryopreservation has been widely used in assisted reproductive technology, as it offers great potential for the treatment of some types of male infertility. However, cryopreservation may result in changes in membrane lipid composition and acrosome status, as well as reductions in sperm motility and viability. This study aimed to evaluate sperm DNA fragmentation damage caused by conventional freezing using the sperm chromatin dispersion test. Methods: In total, 120 fresh human semen samples were frozen by conventional methods, using SpermFreeze Solution as a cryoprotectant. Routine semen analysis and a Halosperm test (using the Halosperm kit) were performed on each sample before freezing and after thawing. Semen parameters and sperm DNA fragmentation were compared between these groups. Results: There was a significant decrease in sperm progressive motility, viability, and normal morphology after conventional freezing (32.78%, 79.58%, and 3.87% vs. 16%, 55.99%, and 2.55%, respectively). The sperm head, midpiece, and tail defect rate increased slightly after freezing. Furthermore, the DNA fragmentation index (DFI) was significantly higher after thawing than before freezing (19.21% prior to freezing vs. 22.23% after thawing). Significant increases in the DFI after cryopreservation were observed in samples with both normal and abnormal motility and morphology, as well as in those with normal viability. Conclusion: Conventional freezing seems to damage some sperm parameters, in particular causing a reduction in sperm DNA integrity.

Antioxidant Effect of Garlic Supplement against Exercise-Induced Oxidative Stress in Rats (운동으로 유발된 산화 스트레스와 마늘의 항산화 작용)

  • Yoon, Gun-Ae
    • Journal of Nutrition and Health
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    • v.40 no.8
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    • pp.701-707
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    • 2007
  • This study was to investigate lipid peroxidation, antioxidant enzyme activity and DNA damage after exercise, and the protective effect of garlic against exercise-induced oxidative stress. Male Sprague-Dawley rats(4 weeks old) were randomly divided into three groups of 6 rats each; control group(Con) without garlic and exercise, Ex group with exercise alone, and Ex-G group with 2% garlic and exercise. For 4 weeks, rats were given diets containing 15% corn oil and 1% cholesterol with or without garlic. The swimming was selected as a model for exercise performance. Rats swam for 40 min a day, for 5 days a week. Group Ex and Ex-G showed significant lowering in body weight gain and fat accumulation compared to control. No significant changes were observed in levels of plasma cholesterol and triglyceride among three groups, demonstrating that exercise and garlic had no effects on changes of blood lipid. This finding of blood lipid seems to be due to higher plant sterol content in corn oil. The DNA tail moment of lymphocytes showed greater tendency in Ex and Ex-G than in control, but garlic supplements failed to suppress DNA damages. Compared to control, Ex had higher plasma TBARS which was lowered to the control's level in Ex-G with 2% garlic supplementation(p<0.05). Ex-G led to a higher hepatic superoxide dismutase(SOD) activity than control and Ex(p<0.05). Activity of hepatic catalase was also increased in Ex-G, while in Ex it was significantly low(p<0.05). It seemed that TBARS levels were related to the activities of SOD and catalase, and that garlic contributed to increasing the enzyme activities and led to decrease of TBARS. These results demonstrate that lipid peroxidation and DNA damage occur as a consequences of oxidative stress after exercise, and that antioxidant defense against oxidative stress could be enhanced by garlic supplementation through the induction of antioxidant enzymes. However, further investigations should be done on the garlic effect on DNA damage.

Characterization of a Low Molecular Weight Heat-Shock Protein cDNA Clone from Nicotiana tabacum

  • Park, Soo-Min;Joe, Myung-Kuk;Hong, Choo-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.04a
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    • pp.18-18
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    • 1999
  • We characterized a cDNA clone for a low molecular weight heat-shock protein (LMW HSP) from tobacco named TLHS-l. Nucleotide sequence determination of TLHS-1 identified an open reading frame for 159 amino acids. To the upstream of the open reading frame, a sequence of 124 nucleotides was determined. To the 3' downstream of the open reading frame, 212 nucleotides were identified which carried poly(A)-tail. Comparison of the open reading frame and hydropathy plot of TLHS-1 with the previously reported class I LMW HSPs showed high identity which classified TLHS-1 as a class I LMW HSP cDNA clone. We proposed that there are six consensus regions in class I LMW HSPs. RNA blot hybridization for TLHS-1 showed a typical expression pattern of heat-shock-inducible gene from three common tobacco cultivars. The open reading frame of TLHS-1 was overexpressed in Escherichia coli. TLHS-1 protein confers thermal protection of other proteins in vitro and in vivo. Thermal induced aggregation of citrate synthase was reduced by purified TLHS-1 protein, and thermal death rate at $50^{\circ}C$ was reduced in E. coli expressing TLHS-l. From these data, we can expect that TLHS-1 acts as a molecular chaperone.perone.

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