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Molecular Cloning and Sequencing of the Ecdysteroid UDP-Glucosyl-transferase Gene, EGT, from Bombyx mori Nuclear Polyhedrosis Virus K1

  • Park, Hye-Jin;Chung, Eun-Hwa;Lee, Kwang-Sik;Han, Ji-Hee;Lee, Seong-Jin;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.37-41
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    • 2001
  • The ecdysteroid UDP-glucosyltransferase (egt) gene isolated from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain was compared to its homologue from Autographa californica NPV (AcNPV) and Bm NPV T3. The egt gene of BmNPV-K1 encoded 506 amino acid open reading frame, and was 99.6% identical at the amino acid level and 99.2% identical at the nucleotide level to BmNPV T3. The BmNPV-K1 egt gene showed highly identity to AcNPV and BmNPV T3 strain. The BmNPV-K1 egt gene was different from amino acid sequence at 2 positions, 19 and 72, in BmNPV T3. The genomic location of egt gene in the BmNPV-K1 was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Northern hybridization analysis. Transcripts of the egt of Bm NPV-K1 peaked around 12 hrs postinfection (p.i.) and reduced at 24 hrs p.i.

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Wide phenotypic variations in Charcot-Marie-Tooth 1A neuropathy with rare copy number variations on 17p12

  • Kanwal, Sumaira;Choi, Byung-Ok;Kim, Sang-Beom;Koo, Hea-Soo;Kim, Jee-Young;Hyun, Young-Se;Lee, Hye-Jin;Chung, Ki-Wha
    • Animal cells and systems
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    • v.15 no.4
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    • pp.301-309
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    • 2011
  • Charcot-Marie-Tooth disease (CMT) is clinically heterogeneous hereditary motor and sensory neuropathies with genetic heterogeneity, age-dependent penetrance, and variable expressivity. Rare copy number variations by nonrecurrent rearrangements have recently been suggested to be associated with Charcot-Marie-Tooth 1A (CMT1A) neuropathy. In our previous study, we found three Korean CMT1A families with rare copy number variations (CNVs) on 17p12 by nonrecurrent rearrangement. Careful clinical examinations were performed in all the affected individuals with rare CNVs (n=19), which may be the first full study of a subject from a large CMT1A family with nonrecurrent rearrangement. The clinical phenotype showed no significant difference compared with common CMT1A patients, but with variable phenotypes. In particular, a broad intrafamilial phenotypic spectrum was observed within the same family, which may suggest the existence of a genetic modifier. This study may broaden the understanding of the role of CNVs in the pathogenesis of CMT.

Expression of Coat Color Associated Genes in Korean Brindle Cattle by Microarray Analysis

  • Lee, Hae-Lee;Park, Jae-Hee;Kim, Jong Gug
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.99-107
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    • 2015
  • The aim of the present study was to identify coat color associated genes that are differentially expressed in mature Korean brindle cattle (KBC) with different coat colors and in Hanwoo cows. KBC calves, before and after coat color appearance, were included. Total cellular RNA was isolated from the tail hair cells and used for microarray. The number of expressed coat color associated genes/probes was 5813 in mature KBC and Hanwoo cows. Among the expressed coat color associated genes/probes, 167 genes were the coat color associated genes listed in the Gene card database and 125 genes were the pigment and melanocyte genes listed in the Gene ontology_bovine database. There were 23 genes/probes commonly listed in both databases and their expressions were further studied. Out of the 23 genes/probes, MLPH, PMEL, TYR and TYRP1 genes were expressed at least two fold higher (p<0.01) levels in KBC with brindle color than either Hanwoo or KBC with brown color. TYRP1 expression was 22.96 or 19.89 fold higher (p<0.01) in KBC with brindle color than either Hanwoo or KBC with brown color, respectively, which was the biggest fold difference. The hierarchical clustering analysis indicated that MLPH, PMEL, TYR and TYRP1 were the highly expressed genes in mature cattle. There were only a few genes differentially expressed after coat color appearance in KBC calves. Studies on the regulation and mechanism of gene expression of highly expressed genes would be next steps to better understand coat color determination and to improve brindle coat color appearance in KBC.

Protective Immune Reponses Induced by Non-infectious L-particles of Equine Herpesvirus Type-1: Implication of Cellular Immunity

  • Mohd Lila Mohd Azmi;Field, Hugh-John;Frazer Rixon;Lauchlan, John-Mc
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.11-19
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    • 2002
  • Mice immunized with equine herpesvirus type-1(EHV-1) L-particles skewed a significant increase (p<7.75) in serum antibody titers. Upon a booster dose four weeks lateral antibody titers increased significantly. Interestingly, immunization via intravenous or intramuscular route induced significantly higher (p<0.75) antibody titers. However, mice iummunized with UV-treated L-particles, visions or immunization via intranasal route induced lower antibody titers. Upon challenge inoculation with wildtype EHV-1, our data showed there was a poor correlation between antibody titers and protection against virus replication. Therefore, the role of cell-mediated immunity Inwards protection was investigated. As predicted, the strongest cell-mediated immunity, as measured by delayed-hypersensitivity test, was detected in mice immunized with live virus particles. The magnitude of cell-mediated immune response correlated with the efficacy of L-particles as immunizing agent. The highest efficacy, as indicated in mice immunized via intranasal routed was highly correlated with cell-mediated immunity. A similar phenomenon was also demonstrated in mice immunized intranasally with UV-treated L-particles. However, the degree of protection was reduced when mice immunized intravenously or intramuscularly with UV-treated L-particles. In conclusion, protection conferred in these animals was highly implicated by immune cells and the least by antibodies. The route of immunization and the nature of the antigen also contributed to the efficacy of L-particles as immunizing agent. In contrast to that of herpes simplex virus type 1, our data showed EHV-1 non-infectious L-particles are highly suitable for immunization of the host against EHV-1 disease.

Production and Reaction Properties of Phytase by Saccharomyces cerevisiae CY strain (Saccharomyces cerevisiae CY 균주에 의한 Phytase의 생성과 반응특성)

  • Seo, Sung-Won;In, Man-Jin;Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.228-232
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    • 2005
  • A yeast strain producing phytase, isolated from a mash of Korean traditional Yakju, was identified as a strain of Saccharomyces cerevisiae and designated as Saccharomyces cerevisiae CY strain. Phytase was produced by CY strain both intracellularly and extracellularly. Total phytase activity by the shaking culture was about two times higher than that of the static culture. The portion of extracellular phytase to total phytase activity ranged between 23 and 49 percent, depending on the glucose concentration in the culture medium. Phytase production was reached at approximately 1 U/ml as total phytase activity and the maximum intracellular phytase activity was 0.17-0.19 U/mg-DCW at late logarithmic growth phase. The optimum reaction pH and temperature of intracellular phytase were 3.5 and $40^{\circ}C$, respectively. Over 95% of the phytate was degraded by growing cells after 36 hours yeast cell culture and about 90% of total phytate was effectively degraded by suspending the whole cell with the biomass of 0.4 mg-DCW/ml-reaction solution after 12 hours degradation reaction.

The Properties of Glucose Isomerase Produced by Streptomyces luteogriseus TH34 (Streptomyces luteogriseus TH34가 생산하는 Glucose Isomerase의 특성)

  • 홍승서;백진기;이현수;국승욱;박관화
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.405-412
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    • 1991
  • The enzymes were immobilized by treating the microbial cells in 0.05% chitosan and 0.28% glutaraldehyde solution. The activity of immobilized cell was about 535 IGIC/g. Glucose isomerase was purified by 6.5 times after homogenization using 60% $(NH_4)_2S0_4$ fractionation, DEAE-cellulose and Sephadex G-150 gel filtration. The molecular weight of enzyme was about 140,000 when it was measured by HPLC and the purified enzyme had only one band by electrophoresis. It showed good enzyme activity at pH 7.5 and $75^{\circ}C$. The optimum conditions for enzyme reactions were shifted to pH 7.0 and $80^{\circ}C$ when the enzyme was immobilized. The enzyme reaction was activated by the addition of 5~10 mM magnesium ion and the thermostability was improved by the addition of 0.25 mM cobalt ion. The enzyme activity was competitively inhibited by sugar alcohols.

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The Effects of Tongkwansan on the Changes of Granulocytes and Nasal Tissue in Allergic Rhinitis Mouse Model (통관산(通關散)이 알레르기성 비염 동물 모델의 과립구 및 조직학적 변화에 미치는 영향)

  • Park, Jong-Won;Nam, Hae-Jeong;Kim, Yoon-Bum
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.19 no.3 s.31
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    • pp.75-89
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    • 2006
  • Background & Objectives : Rhinitis is an inflammation of nasal mucosa and the symtoms are watery rhinorrhea, sneezing, itchy nose, and nasal obstruction. Rhinitis is classified into allergic rhinitis and nonallergic. Allergic rhinitis is an immune reaction by allergen, and vasomotor rhinitis which is nonallergic noninfectious hypersensitive reaction. The incidence of allergic rhinitis has increased and the rate of vasomotor rhinitis is high. However there have been no studies about vasomotor rhinitis compared with allergic rhinitis. And there have been no studies so far performed on the effect of Tongkwansan. Therefore this study is aimed to find out the effects of Tongkwansan on allergic rhinitis and vasomotor rhinitis. Materials and Methods : Fifteen BALC/c mouses divided into three groups : normal group, control group and sample group. To induce the allergic rhinitis in control group and sample group, mouses were sensitized intrapertioneally 0.1 % ovalvumin solution three times at intervals of 1 week. Then intranasal sensitization was performed by diffusing 0.1 % ovalbumin solution 3 times at intervals of 2 days. After that time, mouses in the sample group were oral and administration treated by Tongkwansan for 28 days. We observed changes in nasal mucosa and submucosa; also changes in the segment of leucocyte, erythrocyte, neutrophil, lymphocyte, monocyte, eosinophil, IL-4 and $IFN-{\gamma}$ in blood. We used the statistical methods of ANOVA test(p<0.05). Results : There were no significant changes statistically in leucocyte, erythrocyte, neutrophil, lymphocytem, monocyte, eosinophil, IL-4 and $IFN-{\gamma}$ in blood(p<0.05). Hypertrophy of epithelium in nasal mucosa and expansion of glandular cells in nasal submucosa were decreased in treated group when compared with control group. Conclusion : According to above results, it is supposed that Tongkwansan has significant effects on vasomotor rhinitis which is nonallergic and noninfectious.

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Effects of Feeding Nattokinase as Natural Feed Additives on Milk Production and Blood Metabolites in Lactating Dairy Cows (천연 사료첨가제 Nattokinase 공급에 따른 젖소의 산유능력 및 혈액성상에 미치는 영향)

  • Lim, Dong-Hyun;Park, Joong-Kook;Kim, Hyeon-Shup;Ki, Kwang-Seok;Lee, Hyun-June;Kwon, Eung-Gi;Kim, Mi-Kyoung;Kim, Chang-Hyun;Kim, Sang-Bum
    • Korean Journal of Organic Agriculture
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    • v.19 no.4
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    • pp.553-563
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    • 2011
  • This experiment was conducted to determine the effect of nattokinase (NK) additives on milk production and composition, and blood metabolites in dairy cows. The two kinds of nattokinase with high fibrinolytic activity were produced by two strains of bacteria, Bacillus amyloliquefacines (NK1) and Bacillus subtilis (NK2). Total fifteen Holstein cows (average $1.83{\pm}0.37$ parity; average milk yield $23.2{\pm}3.2$ kg/d) were randomly assigned to three treatments (5 animals per treatment). Cows were fed TMR supplemented with 0g, 100g and 100g for control, NK1 and NK2 treatment, respectively for 4 weeks. Milk yield was significantly higher (p<0.05) for NK1 (22.89 kg/d) than for control (21.07 kg/d) and NK2 (21.36 kg/d). Somatic cell counts in NK treatments were significantly lower than that in control group (58,000 vs. 21,000 and 35,000 cells/ml, control vs. NK1 and NK2). Serum ALT levels in all treatment were similar to the range of 32.00~35.83 IU/L, but AST levels in NK1 (85.67 IU/L) was significantly decreased compared with those in control and NK2 (121.67 and 117.67 IU/L respectively). Serum T-CHO levels in NK1 (145.33 mg/dl) was significantly decreased (p<0.05) compared with that in control (179.00 mg/dl) and NK2 (176.17 mg/dl). This finding showed that NK1 additives could possibly have a positive effect in lactation performance of mid-lactation dairy cows by increasing milk yield, reducing somatic cell count, improving liver function and decreasing cholesterol in blood.

Anti-inflammatory activities of a herbal preparation (HemoHIM) in colitis induced by trinitrobenzene sulfonic acid in rats (Trinitrobenzene sulfonic acid에 의해 유발된 랫드의 대장염에서 HemoHIM의 항염증 효과)

  • Lee, Hae-June;Kim, Se-Ra;Moon, Chang-Jong;Kim, Jong-Choon;Bae, Chun-Sik;Kang, Seong-Soo;Jung, U-Hee;Park, Hae-Ran;Jo, Sung-Kee;Kim, Sung-Ho
    • Korean Journal of Veterinary Research
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    • v.47 no.1
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    • pp.19-24
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    • 2007
  • The cause and pathogenesis of inflammatory bowel disease remain unknown and no definitetherapy exists until now. The present study was conducted to investigate the anti-inflammatory effectsof a herbal preparation (HemoHIM) in colitis induced by 30 mg of trinitrobenzene sulfonic acid (TNBS)in rats. Sprague-Dawley rats were divided into 5 groups. Each group was treated with 1 mg ofHemoHIM/ml of drinking water, 4 mg of HemoHIM/ml of drinking water, 50 mg of HemoHIM/kgof body weight (i.p. once every other day) or 10 mg/kg of HemoHIMof body weight (i.p. onceevery other day) from the next day. After 2 weeks, rats were sacrificed and morphologic featuresof colons were examined. Ulceration, adhesion, thickening and dilatation were noticed in the colonicmucosa after TNBS instillation. Intraperitoneal injection of HemoHIM (50 and 100 mg/kg of bodyweight) showed the anti-inflammatory effect on adhesion, thickening, dilatation, ulceration, and theinhibition effect on damage score by 72.7% and 90.9%, respectively. Histologically, the colon of TNBS-treated rat showed inflammatory cell infiltration by polymorphonuclear cells, multiple erosive lesionsignificant improvement in these symptoms. The results obtained suggest marked anti-inflamatoryactivity of the HemoHIM at the dose levels examined.

Long Noncoding RNA MHRT Protects Cardiomyocytes against H2O2-Induced Apoptosis

  • Zhang, Jianying;Gao, Caihua;Meng, Meijuan;Tang, Hongxia
    • Biomolecules & Therapeutics
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    • v.24 no.1
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    • pp.19-24
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    • 2016
  • Acute myocardial infarction (AMI) remains a leading cause of morbidity and mortality worldwide. The exploration of new biomarkers with high sensitivity and specificity for early diagnosis of AMI therefore becomes one of the primary task. In the current study, we aim to detect whether there is any heart specific long noncoding RNA (lncRNA) releasing into the circulation during AMI, and explore its function in the neonatal rat cardiac myocytes injury induced by $H_2O_2$. Our results revealed that the cardiac-specific lncRNA MHRT (Myosin Heavy Chain Associated RNA Transcripts) was significantly elevated in the blood from AMI patients compared with the healthy control ($^*p<0.05$). Using an in vitro neonatal rat cardiac myocytes injury model, we demonstrated that lncRNA MHRT was upregulated in the cardiac myocytes after treatment with hydrogen peroxide ($H_2O_2$) via real-time RT-PCR (qRT-PCR). Furthermore, we knockdowned the MHRT gene by siRNA to confirm its roles in the $H_2O_2$-induced cardiac cell apoptosis, and found that knockdown of MHRT led to significant more apoptotic cells than the non-target control ($^{**}p<0.01$), indicating that the lncRNA MHRT is a protective factor for cardiomyocyte and the plasma concentration of MHRT may serve as a biomarker for myocardial infarction diagnosis in humans AMI.