• Title/Summary/Keyword: DNA in tail

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Candidate Genes with Ovulation by Differential Display PCR in Small Tail Han Sheep

  • Liu, Shufang;Li, Hongbin;Song, Xuemei;Wang, Aihua;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1229-1233
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    • 2006
  • To find the candidate genes concerned with ovulation rate of sheep, Differential Display Reverse Transcription Polymerase Chain Reaction was employed to find the differently expressed cDNA controlling ovulation in the Small Tail Han sheep of polyembryony and in Tan sheep of single birth. Twenty-four primer pairs of three anchored primers and eight arbitrary primers were assembled to amplify the specialized bands from these sheep. Positive cross tests were applied to optimize the ascertainable PCR conditions in which different special bands can be identified by silver strain in one PCR tube. After eliminating the false positive PCR products by Northern hybridization, 24 differential display bands were acquired from the ovary in the Small Tail Han sheep. These EST bands were sequenced and 18 different ESTs were found in which five ESTs had several copies and 13 ESTs had only one copy. Comparing these ESTs with homologous sequences by BLAST in the GenBank, there were six ESTs with known open reading frame (ORF) and function, three ESTs with known ORF and no function, and 9 ESTs without homologous sequence. These ESTs partly represent several genes such as NOS2, tensin, TCRA, CDKN1A, ESR1 and ACTB which express especially in Small Tail Han sheep.

The Improvement of Chaga Mushroom (Inonotus Obliquus) Extract Supplementation on the Blood Glucose and Cellular DNA Damage in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발한 당뇨쥐에 있어서 차가버섯(Inonotus Obliquus)의 혈당 및 DNA 손상 개선효과)

  • Park, Yoo-Kyoung;Kim, Jung-Shin;Jeon, Eun-Jae;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.42 no.1
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    • pp.5-13
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    • 2009
  • Mushrooms have become a largely untapped source of powerful new pharmaceutical products that poses anti-inflammatory, and antimutagenic, and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protecting cellular components against free radical. The aim of this study was to investigate the protective effect of chaga mushroom against diabetes, via the mitigation of oxidative stress and reduction of blood glucose, in streptozotocin-induced diabetic rats. Rats were rendered diabetic by intravenous administration of STZ through tail at a dose of 50 mg/kg. Animals were allocated into four groups with 8 rats each. The control and diabetic control group were fed with standard rat feed. The other diabeic groups, the low chaga extract group and the high chaga extract group were fed ad libitum using 0.5 g/kg and 5 g/kg of chaga mushroom extract, respectively, for 4 weeks. The blood glucose levels in the two chaga extract groups showed a tendency to decrease but did not reach statistical significance after the supplementation. Leukocyte DNA damage, expressed as tail length, was found to be significantly lower in the high chaga extract group than in the diabetic control group (p > 0.05). Plasma level of total radical-trapping antioxidant potential (TRAP) was tend to be higher in the high chaga extract group compared with the diabetic control group. Erythrocyte antioxidant enzyme activities of two groups did not differ. Although we did not obtain beneficial effect on lowering blood glucose levels in the STZ-induced diabetic rats, this results suggest that the chaga mushroom extracts may initially act on protecting endogenous DNA damage in the short-term experiment.

Characterization of an Easter Lily Calmodulin cDNA Clone (백합실물에서 하나으 Calmodulin cDNA 클론 연구)

  • Kim, Seong-Ryong;An, Gyu-Heung
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.9-13
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    • 1996
  • A clone, LCM1, which encodes calmodulin (CaM) was isolated and characterized from monocot lily (Lilium longiflorum Thunb.) plants. The clone is 681 bps and contains the 447 bp coding region, 8 bp leader sequence, 210 bp 3'-untraslated region, and a poly(A) tail. The coding region of 149 amino acids encodes a protein of predicted Mr 17 kD. Comparison of the LCM1 amino acid sequence with other CaMs revealed that the protein is highly conserved among various living organisms. The expression level of calmodulin gene in lily was studied by RNA blot analysis. The LCM1 mRNA was present in all tissues tested. However, a higher level of calmodulin was observed in anther and floral bud. The level of calmodulin mRNA in anther was about 10 times higher than that in anther was about 10 times higher than that in vegetative tissues. The anther preferential expression of CaM in lily is currently investigated in dicot plants.

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Morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae from Chinese pipistrelle bats and viviparid snails in Thailand

  • Thitichai Arttra;Pheravut Wongsawad;Chalobol Wongsawad;Nattawadee Nantarat;Preeyaporn Butboonchoo;Jong-Yil Chai
    • Parasites, Hosts and Diseases
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    • v.62 no.1
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    • pp.85-97
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    • 2024
  • This study aimed to describe the morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae isolated in Thailand. Adult flukes were isolated from the Chinese pipistrelle bat (Hypsugo sp.), and cercariae were detected in the viviparid snail (Filopaludina martensi martensi) from Chiang Mai province. The morphological characteristics were observed and described using conventional methods, and the molecular characteristics with internal transcribed spacer 2 (ITS2) and 28S rDNA gene sequences. The adult flukes were fusiform, 0.84-0.98 mm in length, and 0.37-0.49 mm in width, and were distinguishable from other species by the presence of longitudinal uterine coils. The cercariae were nonvirgulate xiphidiocercariae, with the oral sucker bigger than the acetabulum, the tail without fin fold, a body size of 117.5-138.3×48.3-52.2 ㎛, and a tail size of 100.7-103.7×15.0-18.9 ㎛. Molecular studies revealed that the adults and cercariae shared 99.3% (ITS2) and 99.6% (28S rDNA) homology with each other. They were phylogenetically close to P. longiforme with an identity of 94.5% for ITS2 and 98.7% for 28S rDNA. This study provides new information on the natural definitive host and first intermediate host of P. longiforme in Thailand. The discovery of its cercarial stage in Filopaludina snails highlights the importance of monitoring the associated second intermediate host and prevention and control of this potentially zoonotic trematode.

Non-histone protein HMGB1 inhibits the repair of damaged DNA by cisplatin in NIH-3T3 murine fibroblasts

  • Yusein-Myashkova, Shazie;Ugrinova, Iva;Pasheva, Evdokia
    • BMB Reports
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    • v.49 no.2
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    • pp.99-104
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    • 2016
  • The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

Isolation and Characterization of UV-inducible gene in Eukaryotic cells

  • Choi, In-Soon
    • Journal of Life Science
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    • v.11 no.1
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    • pp.52-56
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    • 2001
  • The present study intends to characterize the DNA damage-inducible responses in eukaryotic cells. The fission yeast, S. pombe, which displays efficient DNA repair systems, was used in this study as a model system for higher eukaryotes. To study UV-inducible responses in S. pombe, five UV-inducible cDNA clones were isolated from S. pombe by using subtration hybridization method. To investigate the expression of isolated genes, the cellular levels of the transcripts of these genes were determined by Northern blot analysis after UV-irradiation. The transcripts of isolated gene (UV130) increased rapidly and reached maximum accumulation after UV-irradiation. Compared to the message levels of control, the levels of maximal increase were approximately 5 fold to UV-irradiation. In order to investigation whether the increase of UV130 transcripts was a specific results of UV-irradiation, UV130 transcript levels were examined after treating the cells to Methylmethane sulfonate (MMS). The transcripts of UV130 were not induced by treatment of 0.25% MMS. These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of these genes. To characterize the structure of UV130 gene, nucleotide sequences were analyzed. The nucleotide sequence of 1,340 nucleotide excluding poly(A) tail contains one open reading frame, which encodes a protein of 270 amino acids. The predicted amino acid sequences of UV130 do not exhibit any significant similarity to ther known sequences in the database.

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Structural and Functional Insight into Proliferating Cell Nuclear Antigen

  • Park, So Young;Jeong, Mi Suk;Han, Chang Woo;Yu, Hak Sun;Jang, Se Bok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.637-647
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    • 2016
  • Proliferating cell nuclear antigen (PCNA) is a critical eukaryotic replication accessory factor that supports DNA binding in DNA processing, such as DNA replication, repair, and recombination. PCNA consists of three toroidal-shaped monomers that encircle double-stranded DNA. The diverse functions of PCNA may be regulated by its interactions with partner proteins. Many of the PCNA partner proteins generally have a conserved PCNA-interacting peptide (PIP) motif, located at the N- or C- terminal region. The PIP motif forms a 310 helix that enters into the hydrophobic groove produced by an interdomain-connecting loop, a central loop, and a C-terminal tail in the PCNA. Post-translational modification of PCNA also plays a critical role in regulation of its function and binding partner proteins. Structural and biochemical studies of PCNA-protein will be useful in designing therapeutic agents, as well as estimating the outcome of anticancer drug development. This review summarizes the characterization of eukaryotic PCNA in relation to the protein structures, functions, and modifications, and interaction with proteins.

Assessment of Nucleus-DNA Damage in Red Pepper Cells Treated with γ-Radiation through Comet Assay (Comet 분석을 통한 방사선처리 고추세포의 핵 DNA 손상평가)

  • An, Jung-Hee;Back, Myung-Hwa;Kim, Jae-Sung;Jeong, Jeong-Hag;Kwon, Soon-Tae
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.225-230
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    • 2004
  • We employed single cell gel electrophoresis method (comet assay) to analyze the degree of nucleus-DNA damage in the leaves of red pepper (Capsicum annuum L.) seedlings exposed to $^{60}$ CO v-radiation stress. Nucleus-DNA damage was measured as the ratio of tail length (T) to head length (H) in individual comet image isolated from pepper leaf cell. The T/H ratio of control-cells and treated-cells at 50 or 100 Gy were 1.28 and 3.54 or 3.39, respectively, suggesting that nuclei of pepper cells were severely damaged in the integrity of DNA strand by the treatment of enhanced v-radiation. The percentage of head-DNA in control-cells was 76.8%, whereas those of 50 and 100 Gy treated-cells were 55.9% and 59.9%, respectively. Pretreatment of low dose (4 to 20 Gy) radiation to seeds decreased DNA-damage in the leaves of seedlings treated with high dose radiation at 50 or 100 Gy. In this experiment, we developed a sensitive, reliable and rapid method for evaluating genotoxic effect in the nuclei of plant cells by employing comet assay.

Four months of magnetized water supplementation improves glycemic control, antioxidant status, and cellualr DNA damage in db/db mice (제2형 당뇨 모델 db/db 마우스에서 4개월의 자화수 섭취 후 혈당, 항산화 상태 및 세포 DNA 손상 개선 효과)

  • Lee, Hye-Jin;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.49 no.6
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    • pp.401-410
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    • 2016
  • Purpose: Water is magnetically charged upon contact with a magnet. Although magnetic water products have been promoted since the 1930's, they have not received wide acceptance since their effectiveness is still in question; however, some have reported their therapeutic effects on the body, especially the digestive, nervous, and urinary systems. Methods: In this study, the effect of magnetized water on glycemic control of 14 diabetic mice (CB57BK/KsJ-db/db) in comparison with 10 control mice (CB57BK/KsJ-db/+(db/+)) was investigated. Seven diabetic control (DMC) mice and seven diabetic mice + magnetized water (DM+MW) were kept for 16 weeks, followed by intraperitoneal glucose tolerance test (IPGTT). Weekly blood glucose was measured from tail veins. Blood obtained from heart puncture was used for HbA1c analysis. Results: Blood glucose level showed a significant difference starting from the $10^{th}$ week of study ($496.1{\pm}10.2mg/dl$ in DMC vs. $437.9{\pm}76.9mg/dl$ in DM+MW). Blood glucose followed by IPGTT showed no significant difference between groups at 0, 30, 60, 90, and 120 min, although glucose level at 180 min was significantly reduced in DM+MW mice. Plasma insulin level in DM+MW groups was only 39.5% of that of DMC groups ($5.97{\pm}1.69ng/ml$ in DMC vs. $2.36{\pm}0.94ng/ml$ in DM+MW). Levels of HbA1c were 12.4% and 9.7% in DMC and DM+MW groups, respectively. Conclusion: These results show the promising therapeutic effect of magnetized water in regulating blood glucose homeostasis; however, long-term supplementation or mechanistic study is necessary.

The Protective Effects of Ethanol Extract of Wild Simulated Ginseng on Carbon Tetrachloride Induced Acute Hepatic Injury in Mouse (사염화탄소 유발 급성 간독성 생쥐모델에서 산양삼 에탄올 추출물의 간 보호 효과)

  • Lee, Soo-Min;Park, Sun-Young;Jang, Gi-Seuk;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.41 no.8
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    • pp.701-710
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    • 2008
  • The wild simulated ginseng (WSG) has been effectively used in folk medicine as a remedy against hepatic disease, hypertension and arthritic disease. However, there is still lack of scientific proof about its antioxidant capability. The present study has been conducted to evaluate the protective role of the WSG ethanol extract in the CCl4-induced oxidative stress and resultant hepatic disfunction in ICR mice. The electron donating abilities and IC50 of WSG etnanol extract were 76.86 ${\pm}$ 1.06% and 33.3 ${\mu}g$/mL (that of ascobic acid was 16.5 ${\mu}g$/mL), respectively. Total antioxidant status of WSG extract was 2.13 ${\pm}$ 0.06 mmoL/mg, while the values of ascorbic acid and BHT were 3.63 ${\pm}$ 0.06 and 3.12 ${\pm}$ 0.02, respectively. ICR mice (aged 3weeks) were fed for 4 weeks on AIN-93M diet and had free access to food and water. The animals were divided into three groups: normal group (intraperitoneally (i.p) injected with PBS at 100 ${\mu}L$/mouse), group C; CCl4-induced and without any treatment. (i.p injected only PBS, 100 ${\mu}L$ /mice), group G; CCl4-induced and treated with WSG (i.p injected with 5 mg WSG extract per mouse, suspended in 100 ${\mu}L$ phosphate buffer). After the i.p. injection of WSG or PBS (5 times for 7weeks), all mice were administered CCl4 in olive oil at the last day of the experiment, except for normal group. The normal group was administered only olive oil. Determination of plasma triglyceride, total cholersterol, fasting glucose and GPT activity was performed using automatic blood analyzer. To evaluate the protective effect against the oxidative stress, DNA fragmentation and TBARS were determined in blood leucocytes and RBC and hepatocyte, respectively. Body and organs weights and food intake did not show significant differences among the groups. Blood total cholesterol of group G was similar to that of normal group, which was the lowest in group C. The fasting blood glucose level was the highest in normal group (205.20 ${\pm}$ 135.24), which were decreased in group C (134.2 ${\pm}$ 79.31) and group G (126.48 ${\pm}$ 77.05). TBARS values in a red blood cell and hepatic tisuue homogenate were lower in group G comparing to the group C. DNA% in tail, tail length (TL) and tail moment (TM) of blood leucoocytes showed the highest values in group C (20.11 ${\pm}$ 2.47, 17.36 ${\pm}$ 2.58, 94.11 ${\pm}$ 12.29) and they were significantly diminished in group G (9.63 ${\pm}$ 1.19, 7.04 ${\pm}$ 1.50, 38.64 ${\pm}$ 7.60). In conclusion, wild simulated ginseng might be a protective agent against the oxidative stress.