• 제목/요약/키워드: ITS marker

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Polymorphism of Insulin-like Growth Factor-I Gene in 13 Pig Breeds and its Relationship with Pig Growth and Carcass Traits

  • Wang, Wenjun;Huang, Lusheng;Chen, Kefei;Gao, Jun;Ren, Jun;Ai, Huashui;Lin, Wanhua
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.10
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    • pp.1391-1394
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    • 2002
  • The polymorphism of insulin-like growth factor-I (IGF-I) in 13 pig breeds (total n=559) was detected by PCR-Hha I- RFLP, and allele A (151 bp and 28 bp) or allele B (116 bp, 35 bp and 28 bp) were observed. In these pig breeds, it was found that European pig breeds carried high frequencies of allele B, while Chinese native pig breeds carried high frequencies of allele A. Meanwhile the role of porcine IGF-I was investigated in 117 Nanchang White pigs and 360 Large Yorkshire pigs. Eight traits about growth and carcass were recorded for analyzing the associations between IGF-I gene polymorphism and performance quantitative traits. In the Nanchang White pigs, those with AA genotype generally had higher birth weight than those with AB genotype (p<0.05), but all these genotypes had no significant effect on the other traits which had been analyzed. In Large Yorkshire pigs, those with BB genotype had higher 2 months and 6 months body weight than those with AA genotype (p<0.05), and had a thicker hind-back-fat thickness and mid-back-thickness than those with AB and BB genotypes (p<0.05). And those with BB genotype were the thinnest in Large Yorkshire. Furthermore, pigs with AA genotype had a lower lean percentage than those with AB and BB genotypes (p<0.01), and the lean percentage of those with BB genotype was the highest. Based on these results, it is possible to make the IGF-I gene locus into the application of marker-assisted selection programmes.

Smad4 Expression in Hepatocellular Carcinoma Differs by Hepatitis Status

  • Yao, Lei;Li, Fu-Jun;Tang, Zhi-Qiang;Gao, Shuang;Wu, Qe-Quan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1297-1303
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    • 2012
  • Aims: Primary hepatocellular carcinoma (HCC) is a common malignancy often related to hepatitis viral infection. Smad4 is known to mediate the TGF-${\beta}$ pathway to suppress tumorigenesis. However, the function of Smad4 in HCC is still controversial. In this study we compared levels of Smad4 in HCC tissues with or without hepatitis virus infection and adjacent normal-appearing liver. Methods: Samples from HCC patients were analyzed for Smad4 protein and mRNA expression by immunohistochemistry (IHC), RT-PCR and Western blotting. Results: We found that tumor tissues expressed less Smad4 mRNA and protein than the adjacent tissues. Most HCC tumor tissues were negative for Smad4 in IHC staining, while the majority of adjacent tissues were positively stained. Interestingly, protein levels were higher in HCC tissues with viral hepatitis than those without virus infection. Suppression of expression appeared closely related to HCC, so that Smad4 appears to function as a tumor suppressor gene (TSG). Conclusion: Patients with hepatitis viral infection, at higher risk for HCC, exhibited increased Smad4 protein expression suggesting hepatitis virus may modulate Smad4 expression, which is functionally distinct from its putative role as a TSG. Smad4 expression may thus be an applicable marker for diagnosis and/or a target to develop therapeutic agents for HCC.

Diagnostic Value of Fecal Calprotectin as a Screening Biomarker for Gastrointestinal Malignancies

  • Khoshbaten, Manouchehr;Pishahang, Parinaz;Nouri, Mohammad;Lashkari, Alireza;Alizadeh, Mahasti;Rostami-Nejad, Mohammad
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1667-1670
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    • 2014
  • Background: Calprotectin in feces seems to be a more sensitive marker for gastrointestinal (GI) cancers than fecal occult blood, but its specificity may be too low for screening average risk populations. This study aims at evaluating the diagnostic value of fecal calprotectin as a screening biomarker for GI malignancies. Materials and Methods: In a case-control study, 100 patients with GI malignancies (50 patients with colorectal cancer and 50 patients with gastric cancer) and 50 controls were recruited in Tabriz Imam Reza and Sina hospitals during a 24-month period. One to two weeks after the last endoscopy/colonoscopy, fecal specimens were collected by the patients and examined by ELISA method for quantitative measurement of calprotectin content. The results were compared between the three groups. Results: The mean fecal calprotectin level was $109.1{\pm}105.3$ (2.3-454.3, median:74), $241.1{\pm}205.2$ (3.4-610.0, median:19.3) and $45.9{\pm}55.1{\mu}g/g$ (1.3-257.1, median:19.3) in gastric cancer, colorectal cancer and control group, respectively, the differences being significant (p<0.001) and remaining after adjustment for age. The optimal cut-off point for fecal calprotectin was ${\geq}75.8{\mu}g/g$ for distinguishing colorectal cancer from normal cases (sensitivity and specificity of 80% and 84%, respectively). This value was ${\geq}41.9{\mu}g/g$ for distinguishing gastric cancer from normal cases (sensitivity and specificity of 62%). Conclusions: Our results revealed that fecal calprotectin might be a useful and non-invasive biomarker for distinguishing colorectal cancer from non-malignant GI conditions. However, due to low sensitivity and specificity, this biomarker may not help physicians distinguishing gastric cancer cases from healthy subjects.

Influence of Adipocytokines and Periprostatic Adiposity Measurement Parameters on Prostate Cancer Aggressiveness

  • Zhang, Qiang;Sun, Li-Jiang;Qi, Jun;Yang, Zhi-Gang;Huang, Tao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1879-1883
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    • 2014
  • Background: The relationship between obesity and prostate cancer aggressiveness is controversial in recent studies, partly because BMI is the only generally applied marker of obesity. Our study aimed at evaluating the correlation of periprostatic fat (PF) on magnatic resonance imaging (MRI) and adipocytokines with prostate cancer aggressiveness. Patients and method: A total of 184 patients who underwent radical retropubic prostatectomy (RRP) were analyzed retrospectively; different fat measurements on MRI slices and levels of adipocytokines were compared with the clinical and pathologic factors using SSPS ver.13.0. Result: The PF rates showed a statistically significant variation (p=0.019, 0.025) among groups, that is to say, more adipose tissue was distributed in periprostatic areas of high risk patients. Logistic regression analysis adjusted for age revealed a statistically association between the PF, the ratio and the risk of having high-risk disease (p=0.031, 0.024). The levels of IL-6, leptin and c-reactive protein (CRP) significantly increased with the aggressiveness of prostate cancer, and also with PF and its ratio. The strongest correlation was seen between IL-6 and PF (Pearson r coefficient=0.67, P<0.001). No association was observed between adipocytokines and BMI. Conclusion: Periprostatic adiposity not only affects prostate cancer aggressiveness, but also influences the secretion of adipocytokines. IL-6, PF and CRP have promoting effects on progression of prostate cancer.

Quantitative Determination of Compounds from Akebia quinata by High-Performance Liquid Chromatography

  • Yen, Nguyen Thi;Thu, Nguyen Van;Zhao, Bing Tian;Lee, Jae Hyun;Kim, Jeong Ah;Son, Jong Keun;Choi, Jae Sui;Woo, Eun Rhan;Woo, Mi Hee;Min, Byung Sun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.7
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    • pp.1956-1964
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    • 2014
  • To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D (1), 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2), 3-caffeoylquinic acid (3) and calceolarioside B (4). Particular attention was focused on the main compound, 3-caffeoylquinic acid (3), which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2) was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

Acute toxicity evaluation of drought-tolerant transgenic rice Agb0103 to Daphnia magna

  • Oh, Sung-Dug;Lee, Bum Kyu;Park, Soo-Yun;Yun, Doh-Won;Sohn, Soo-In;Chang, Ancheol;Suh, Sang Jae
    • Korean Journal of Agricultural Science
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    • v.43 no.2
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    • pp.205-214
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    • 2016
  • A drought-tolerant transgenic rice (Agb0103) was developed using a pepper methionine sulfoxide reductase (CaMsrB2) under the control of rice Rab21 promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. Commercialization of genetically modified (GM) crops will require the evaluation of risks associated with the release of GM crops. With the potential problems associated to GM crops safety testing, the investigation of their effects on non-target organisms is necessary for environmental risk research. This study was carried out to assess acute toxicity of a GM crop using the water flea (Daphnia magna) for non-target organism risk evaluation. The effect of acute toxicity on Daphnia magna of Agb0103 rice and a non-GM rice, Ilmibyeo, were investigated at different concentrations (0, 625, 1,250, 2,500, 5,000, and 10,000 mg/L). The Agb0103 rice used for the test was confirmed to express the CaMsrB2/PAT gene by the PCR and ELISA. Daphnia magna feeding tests showed no significant differences in cumulative immobility or abnormal response with either Agb0103 rice or non-GM rice. The 48hr-EC50 values showed no difference between Agb0103 rice (2243 mg/L) and non-GM rice (2694 mg/L). These results suggest that there is no significant difference in toxicity to Daphnia magna between Agb0103 rice and its non-GM counterpart.

Phenotypic and genotypic screening of rice accessions for salt tolerance

  • Reddy, Inja Naga Bheema Lingeswar;Kim, Sung-Mi;Yoon, In Sun;Kim, Beom-Gi;Kwon, Taek-Ryoun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.188-188
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    • 2017
  • Rice (Oryza sativa L.) is one of the major crops that is seriously impacted by global soil salinization. Rice is among those crops where most of the high-yielding cultivars are highly sensitive to salinity. The key to a plant survival under NaCl salt stress is by maintaining a high $K^+/Na^+$ ratio in its cells. Selection for salinity tolerance genotypes of rice based on phenotypic performance alone is less reliable and will delay in progress in breeding. Recent advent of molecular markers, microsatellites or simple sequence repeats (SSRs) were used to find out salt tolerant rice genotypes. In the current experiment phenotyping and genotyping studies were correlated to differentiate different rice accessions for salinity tolerance. Eight rice accessions along with check plant Dongjin were screened by physiological studies using Yoshida solution with 50mM NaCl stress condition. The physiology studies identified four tolerant and four susceptible accessions based on their potassium concentration, sodium concentration, $K^+/Na^+$ ratio and biomass. 17 SSR markers were used to evaluate these rice accessions for salt tolerance out of which five molecular markers were able to discriminate tolerant accessions from the susceptible accessions. Banding pattern of the accessions was scored comparing to the banding pattern of Dongjin. The study identifies accessions based on their association of $K^+/Na^+$ ratio with molecular markers which is very reliable. These markers identified can play a significant role in screening large set of rice accessions for salt tolerance; these markers can be utilized to improve salt tolerance of commercial rice varieties with marker-assisted selection (MAS) approach.

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Identification and molecular characterization of downy mildew resistant gene candidates in maize (Zea mays subsp. Mays)

  • Kim, Jae Yoon;Kim, Chang-Ho;Kim, Kyung Hee;Lee, Byung-Moo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.113-113
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    • 2017
  • Downy mildew (DM), caused by several species in the Peronosclerospora and Scleropthora genera, is a major maize (Zea mays L.) disease in tropical or subtropical regions. DM is an obligate parasite species in the higher plants and spreads by oospores, wind, and mycelium in seed surface, soil, and living hosts. Owing to its geographical distribution and destructive yield reduction, DM is one of the most severe maize diseases among the maize pathogens. Positional cloning in combination with phenotyping is a general approach to identify disease resistant gene candidates in plants; however, it requires several time-consuming steps including population or fine mapping. Therefore, in the present study, we suggest a new combination strategy to improve the identification of disease resistant gene candidates. Downy mildew (DM) resistant maize was selected from five cultivars using the spreader row technique. Positional cloning and bioinformatics tools identified the DM resistant QTL marker (bnlg1702) and 47 protein coding genes annotations. Eventually, 5 DM resistant gene candidates, including bZIP34, Bak1, and Ppr, were identified by quantitative RT-PCR without fine mapping of the bnlg1702 locus. Specifically, we provided DM resistant gene candidates with our new strategy, including field selection by the spreader row technique without population preparation, the DM resistance region identification by positional cloning using bioinformatics tools, and expression level profiling by quantitative RT-PCR without fine mapping. As whole genome information is available for other crops, we propose applying our novel protocol to other crops or for other diseases with suitable adjustment.

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An Assay Method for Screening Inhibitors of Prolyl 4-hydroxylase in Immortalized Rat Hepatic Stellate HSC-T6 Cells

  • Choi, Hwa-Jung;Soh, Yun-Jo
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.261-265
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    • 2007
  • Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase (P4H) which plays a crucial role in the synthesis of all collagens. Considering the role of collagen and its significance in many clinically important diseases such as liver fibrosis, a great deal of attention has been directed toward the development of an assay at cell-based system. The reason is that cell-based assay system is more efficient than enzyme-based in vitro system and takes much less time than in vivo system. Several assay procedures developed for P4H are laborious, time-consuming and not feasible for the massive-screening. Here, we report the cell-based assay method of prolyl 4-hydroxylase in immortalized rat hepatic stellate HSC-T6 cells. To optimize the cell culture condition to assay for HYP content, various concentrations of reagents were treated for different times in HSC-T6 cells. Our data showed that the treatment with ascorbate in a hypoxic condition for 24 h resulted in the maximal increase of HYP by 1.8 fold. Alternatively, cobalt chloride ($5\;{\mu}M$) and ascorbate ($50\;{\mu}M$) in normoxic states exhibited similar effect on the production of HYP as in a hypoxic condition. Therefore, cobalt chloride can be substituted for a hypoxic condition when an anaerobic chamber is not available. Rosiglitazone and HOE077, known as inhibitors of collagen, synthesis decreased P4H enzyme activity by 32.3% and 15%, respectively, which coincided with previous reports from liver tissues. The level of the smooth muscle ${\alpha}$-actin, a marker of activated stellate cells, was significantly increased under hypoxia, suggesting that our experimental condition could work for screening the anti-fibrotic compounds. The assay procedure took only 3 days after treatment with agents, while assays from the primary stellate cells or liver tissues have taken several weeks. Considering the time and expenses, this assay method could be useful to screen the compounds for the inhibitor of prolyl 4-hydroxylase.

High $T_c$ SQUID system for biological immunoassays

  • Enpuku, K.
    • 한국초전도학회:학술대회논문집
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    • v.10
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    • pp.7-7
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    • 2000
  • A high $T_c$ SQUID system is developed for the application to biological immunoassay. In this application, magnetic nanoparticles are used as magnetic markers to perform immunoassay, i.e., to detect binding reaction between an antigen and its antibody. The antibody is labeled with ${\gamma}-Fe_2O_3\;(or\;Fe_3O_4)$ nanoparticles, and the binding reaction can be magnetically detected by measuring the magnetic field from the nanoparticles. Design and set up of the system is described. The system consists of (1) SQUID magnetometer or gradiometer made of 30-deg. bicrystal junctions, (2) field and compensation coils to apply the magnetic field of about 1 mT, (3) special Dewar to realize a 2 mm-distance between the SQUID and the sample, (4) two layers of cylindrical shielding to reduce the extemal magnetic noise to about 1/100, and (5) an electric slider to move the sample with a speed of 10 mm/sec. The sensitivity of the system is studied in terms of detectable magnetic flux. For the measurement bandwidth from 0.2 Hz to 10 Hz, minimum-detectable amplitude of the magnetic flux is $0.8\;m\;{\Phi}_o$ and $0.25\;m{\Phi}_o$ for the magnetometer and the gradiometer, respectively, when the magnetic field of 1 mT is applied. The difference between them is due to the residual environmental noise, and the applied magnetic field does not increase the system noise. The corresponding weight of the magnetic markers is 1 ng and 310 pg, respectively. An experiment is also conducted to measure antigen-antibody reaction with the present system. It is shown that the sensitivity of the present system is 10 times better than that of the conventional method using an optical marker. A one order of magnitude improvement of sensitivity will be realized by the sophistication of the present system.

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