• Title/Summary/Keyword: ITS-PCR

Search Result 2,032, Processing Time 0.03 seconds

Evaluation of the Accuracy of the $EasyTest^{TM}$ Malaria Pf/Pan Ag, a Rapid Diagnostic Test, in Uganda

  • Chong, Chom-Kyu;Cho, Pyo Yun;Na, Byoung-Kuk;Ahn, Seong Kyu;Kim, Jin Su;Lee, Jin-Soo;Lee, Sung-Keun;Han, Eun-Taek;Kim, Hak-Yong;Park, Yun-Kyu;Cha, Seok Ho;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
    • /
    • v.52 no.5
    • /
    • pp.501-505
    • /
    • 2014
  • In recent years, rapid diagnostic tests (RDTs) have been widely used for malaria detection, primarily because of their simple operation, fast results, and straightforward interpretation. The Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag is one of the most commonly used malaria RDTs in several countries, including Korea and India. In this study, we tested the diagnostic performance of this RDT in Uganda to evaluate its usefulness for field diagnosis of malaria in this country. Microscopic and PCR analyses, and the Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag rapid diagnostic test, were performed on blood samples from 185 individuals with suspected malaria in several villages in Uganda. Compared to the microscopic analysis, the sensitivity of the RDT to detect malaria infection was 95.8% and 83.3% for Plasmodium falciparum and non-P. falciparum, respectively. Although the diagnostic sensitivity of the RDT decreased when parasitemia was ${\leq}500\;parasites/{\mu}l$, it showed 96.8% sensitivity (98.4% for P. falciparum and 93.8% for non-P. falciparum) in blood samples with parasitemia ${\geq}100\;parasites/{\mu}l$. The specificity of the RDT was 97.3% for P. falciparum and 97.3% for non-P. falciparum. These results collectively suggest that the accuracy of the Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag makes it an effective point-of-care diagnostic tool for malaria in Uganda.

Comparison of Biological Characteristics on the Organic Waste-treated Lysimeter Soil by RFLP, PLFA, and CLSU (RFLP, PLFA, CLSU를 이용한 폐기물연용토양의 토양미생물 특성 평가 비교)

  • Jang, Kab-Yeul;Weon, Hang-Yeon;Lee, Kang-Hyo;Kwon, Sun-Ik;Kong, Won-sik;Suh, Jang-sun;Sung, Jae-Mo
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.41 no.6
    • /
    • pp.415-418
    • /
    • 2008
  • The application of sludge wastes into agricultural fields has been increasing annually in Korea. In particular, sewage sludge application has been widely accepted in decades. Sewage sludge application aid in the recycling of essential nutrients and act as a source of organic matter improving the structure and water-holding properties of the soil. The efficient use of sludge wastes, however, requires an individual assessment of waste products. This study assessed the biological characteristics of organic waste-treated lysimeter soils and develop its indicator to assess the soil health of organic waste-treated lysimeter soils. Several analytical techniques more recently developed such as restriction fragment length polymorphism (RFLP), phospholipid fatty acid (PLFA), and community level substrate utilization (CLSU) fingerprints allow for detailed analyses of soil microbial communities. PLFA and RFLP was, therefore, used in the study to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. PLFA, RFLP and CLSU have been utilized to assess microbial characteristics of the lysimeter soils with four different sludge wastes for eight consecutive years. Each of these methods was analyzed for a different aspect of soil microbial characteristics. The study would disclose those methods yielded highly reproductive results for each soil and allow distinguishing the soils based on the structures of specific geneand PLFA-pools more than CLSU fingerprints. PLFA methods, especially, revealed the same relative similarities of the treated soils based on cluster analysis of the biological characteristics. Pig manure compost-treated soil, however, was only the same relative resemblance among the three methods. These results indicated that PLFA easily assessed the biological soil characterization.

CLINICAL IMPLICATIONS OF TELOMERASE ACTIVITY IN ORAL SQUAMOUS CELL CARCIMOMA (구강편평세포암에서 telomerase 활성도의 임상적 연관성에 관한 연구)

  • Shim, Yu-Jin;Kim, Myung-Jin;Nahm, Dong-Seok;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.27 no.4
    • /
    • pp.289-300
    • /
    • 2001
  • Telomerase is a ribonucleoprotein that synthesizes telomere repeats. It has been reported that activation of telomerase was associtated with immortalization, proliferative activity and carcinogenesis. Recently, telomerase activity has been extensively studied in many kinds of malignant tumors for clinical diagnostic and/or prognostic utilities. In neuroblastoma, breast carcinoma, gastric carcinoma, non-small cell lung carcinoma, close relationship has been reported between high telomerase activity and lymph node metastasis, tumor aggressiveness and poor prognosis. The purpose of this study is to investigate the clinical implication of telomerase activity assay as an adjunctive factor in decision-making on neck node management, speedy pre-operative judging on histologic malignancy grading. Thus we performed semi-quantitative assay of telomerase activity using Telomerase PCR ELISA $kit^{(R)}$(Boeringer Manheim, Germany) and evaluated correlation between telomerase activity and tumor size, neck node metastasis, Anneroth malignancy score and influence of pre-operative chemotherapy on its activity in 27 cases of oral squamous cell carcinomas and 18 cases of normal oral epithelium. Also, correlation between telomerase activities and PCNA indices was evaluated. The results were obtained as follows: 1. The telomerase activities were detected in 24 specimens out of 27 oral squamous cell carcinoma specimens (88.9%) and in 5 specimens out of 18 normal oral epithelium specimens (27.8%). The mean value of telomerase activities was $0.9793{\pm}0.3428$ in 24 oral squamous cell carcinoma specimens and $0.4855{\pm}0.1117$ in 5 normal oral epithelium specimens. The positivity rate and mean value of telomerase activities in oral squamous cell carcinoma specimens were significantly higher than those of normal oral epithelium specimens (p<0.05). 2. There was no significant correlation between total Anneroth malignancy score and telomerase activity (p>0.05), but points of mitosis index and depth of invasion were significantly correlated with telomerase activities (p<0.05). 3. The positive immunohistochemical staining for PCNA(proliferating cell nuclear antigen) was observed in 26 specimens out of 27 oral squamous cell carcinoma specimens and mean value of PCNA indices of 26 specimens was $53.67{\pm}26.46$. PCNA indices were significantly correlated with telomerase activities (p<0.05). 4. The mean value of telomerase activities was significantly higher in pathologic T3/T4 group than in T1/T2 group (p<0.01). There was no significant difference of mean value of telomerase activities between pathologic neck node positive group and negative group (p> 0.05). Pre-operative chemotherapy significantly lowered the telomerase activities (p<0.05). The above results suggested telomerase activity could be used as diagnostic marker and adjunctive parameter for judging on histologic malignancy in oral squamous cell carcinoma.

  • PDF

Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli (사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구)

  • 이우길;김영호;양중익;노현모
    • Korean Journal of Microbiology
    • /
    • v.28 no.2
    • /
    • pp.91-97
    • /
    • 1990
  • Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

  • PDF

Reidentification of Comamonas sp. Strain DJ-12 and Analysis of its pcbABC2D2 Genes Responsible for Degradation of 4-Chlorobiphenyl. (Comamonas sp. Strain DJ-12 의 재동정 및 4-Chlorobiphenyl 분해유전자 pcbABC2D2 의 분석)

  • 이준훈;박동우;강철희;채종찬;이동훈;김치경
    • Korean Journal of Microbiology
    • /
    • v.40 no.2
    • /
    • pp.121-126
    • /
    • 2004
  • Comamonas sp. strain DJ-12 is a 4-chlobiphenyl(4CB)-degrading bacterium that was reidentified from Pseudomonas sp. DJ-12. The genomic DNA was isolated from the strain DJ-12 and amplified by PCR with primers for cloning pcbABCD genes responsible for degradation of 4CB. The amino acid sequences deduced from the nucleotide sequences of pcbA1, pcbA2, pcbA3, pcbA4, pcbB, pcbC2, and pcbD2 genes showed 91, 87, 99, 87, 97, 90 and 87% homologies with those of Pseudomonas sp. KKS102, respectively. The pcbC1D1 genes that are involved in the degradation of (4-chloro)1,2-dihydroxybiphenyl produced from 4CB by pcbAB gene products were previously reported in the recombinant plasmid pCU1 from Pseudomonas sp. DJ-12. However, the pcbC2D2 genes in the plasmid pCT4 and pCT5 cloned from Comamonas sp. DJ-12 in this study showed 51 and 62% homologies with those of pcbC1D1 in their nucleotide sequences. The pcbC1D1 and pcbC2D2 genes were found by Southern hybridization to be located at different loci on the chromosome of DJ-12 strain. These results indicate that Comamonas sp. strain DJ-12 has two different sets of pcbCD genes responsible for deg-radation of (4-chloro)1,2-dihydroxybiphenyl.

The Study on the Sexual Difference in the Cause and the Time of Casualty and in the Size of the Fairy Pitta (Pitta nympha) through DNA Analysis in Republic of Korea (DNA 분석에 의한 팔색조의 암수 구분 및 암수별 피해 현황 그리고 크기 차이에 관한 연구)

  • Kim, Eun-Mi;Jeon, Yeon-Seon;Jeong, Gil-Sang;Kim, Se-Jae;Kang, Chang-Wan;Oh, Mi-Rea;Noh, Pu-Reum;Won, Hyun-Kyu
    • Journal of Environmental Science International
    • /
    • v.23 no.8
    • /
    • pp.1447-1453
    • /
    • 2014
  • The differentiation of sex is important for species preservation. However, Fairy Pitta is sexually monomorphic and sex of an individual is indistinguishable with its external characteristics. We determined the sex of Fairy Pitta through DNA analysis and investigated the causes and time of injury and mortality and the size based on sex. We collected 21 samples at Jeju Island, Korean Peninsula from 2004 to 2013 and extracted DNA from them and amplified chromo helicase DNA-binding gene from Z and W chromosomes through Polymerase Chain Reaction (PCR). We confirmed their sex with the banding pattern through Agarose gel electrophoresis, i.e. male (ZZ): one banded and female (ZW) two banded. We distinguished the sex of 17 of 21 samples resulting in 9 males and 8 females. Most casualties were recorded in adult of both sexes. Causes of injury and mortality proved that female casualties occurred from window strikes, dehydration, car accident, predation by natural enemies, and male occurred from window strikes, car accident and dehydration. The time of injury and mortality in adults differ by sex. There was no difference between sexes in any of the six size parameters. As the time of injury and mortality differ by sex, the survey on the role and ecological nature by sex in breeding season must be carried out in the future. External measurements may not be reliable for sexing of Fairy Pitta and other traits such as vocal or characteristics are required to identify the sex of individuals in the field.

Bioconverted Jeju Hallabong tangor (Citrus kiyomi × ponkan) peel extracts by cytolase enhance antioxidant and anti-inflammatory capacity in RAW 264.7 cells

  • Chang, Yun-Hee;Seo, Jieun;Song, Eunju;Choi, Hyuk-Joon;Shim, Eugene;Lee, Okhee;Hwang, Jinah
    • Nutrition Research and Practice
    • /
    • v.10 no.2
    • /
    • pp.131-138
    • /
    • 2016
  • BACKGROUND/OBJECTIVES: Citrus and its peels have been used in Asian folk medicine due to abundant flavonoids and usage of citrus peels, which are byproducts from juice and/or jam processing, may be a good strategy. Therefore, the aim of this study was to examine antioxidant and anti-inflammatory effects of bioconversion of Jeju Hallabong tangor (Citrus kiyomi ${\times}$ ponkan; CKP) peels with cytolase (CKP-C) in RAW 264.7 cells. MATERIALS/METHODS: Glycosides of CKP were converted into aglycosides with cytolase treatment. RAW 264.7 cells were pre-treated with 0, 100, or $200{\mu}g/ml$ of citrus peel extracts for 4 h, followed by stimulation with $1{\mu}g/ml$ lipopolysaccharide (LPS) for 8 h. Cell viability, DPPH radical scavenging activity, nitric oxide (NO), and prostagladin $E_2$ ($PGE_2$) production were examined. Real time-PCR and western immunoblotting assay were performed for detection of mRNA and/or protein expression of pro-inflammatory mediators and cytokines, respectively. RESULTS: HPLC analysis showed that treatment of CKP with cytolase resulted in decreased flavanone rutinoside forms (narirutin and hesperidin) and increased flavanone aglycoside forms (naringenin and hesperetin). DPPH scavenging activities were observed in a dose-dependent manner for all of the citrus peel extracts and CKP-C was more potent than intact CKP. All of the citrus peel extracts decreased NO production by inducible nitric oxide synthase (iNOS) activity and $PGE_2$ production by COX-2. Higher dose of CKP and all CKP-C groups significantly decreased mRNA and protein expression of LPS-stimulated iNOS. Only $200{\mu}g/ml$ of CKP-C markedly decreased mRNA and protein expression of cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Both 100 and $200{\mu}g/ml$ of CKP-C notably inhibited mRNA levels of $interleukin-1{\beta}$ ($IL-1{\beta}$) and IL-6, whereas $200{\mu}g/ml$ CKP-C significantly inhibited mRNA levels of $TNF-{\alpha}$. CONCLUSIONS: This result suggests that bioconversion of citrus peels with cytolase may enrich aglycoside flavanones of citrus peels and provide more potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by increasing radical scavenging activity and suppressing pro-inflammatory mediators and cytokines.

Analysis of Molecular Expression in Adipose Tissue-Derived Mesenchymal Stem Cells : Prospects for Use in the Treatment of Intervertebral Disc Degeneration

  • Jin, Eun-Sun;Min, Joongkee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
    • /
    • v.53 no.4
    • /
    • pp.207-212
    • /
    • 2013
  • Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.

Polymorphism of inter simple sequence repeat markers in Hypsizygus marmoreus (Inter Simple Sequence Repeat(ISSR) 마커를 활용한 느티만가닥버섯(Hypsizigus marmoreus) 종내 다형성 분석)

  • Oh, Youn-Lee;Nam, Yun-Geul;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon;Choi, In-Geol
    • Journal of Mushroom
    • /
    • v.15 no.4
    • /
    • pp.273-278
    • /
    • 2017
  • Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

Comparative Analyses for Aroma and Agronomic Traits of Native Rice Cultivars from Central Asia

  • Sarhadi, Wakil Ahmad;Hien, Nguyen Loc;Zanjani, Mehran;Yosofzai, Wahida;Yoshihashi, Tadashi;Hirata, Yutaka
    • Journal of Crop Science and Biotechnology
    • /
    • v.11 no.1
    • /
    • pp.17-22
    • /
    • 2008
  • Aromatic rice has become popular owing to its aroma. Growing demand for aromatic rice has spurred interest in the development of domestic cultivars that offer similar combinations of grain attributes such as texture, cooking characteristics, aroma, and taste. In this study, the most important agronomic attributes and aroma of 26 cultivars from Afghanistan, Iran, and Uzbekistan, and controls from Japan, Thailand, and India were characterized. Also $F_2$ populations derived from the cross between(Jasmine 85 aromatic$\times$Nipponbare non-aromatic) and(Jasmine 85$\times$Basmati 370 aromatic) were obtained. Tasting individual grains, cooking test, 1.7% KOH sensory test, and molecular marker analysis have been applied to distinguish between aromatic and non-aromatic rice. Diversity for some traits of agronomic importance, such as plant height was detected among countries, e.g. Afghan cultivars classified as tall, and Iranian and Uzbek intermediate and short, respectively. Differentiations of panicle, grain, leaf, basal internode, and culm dimension among rice cultivars, indicating the source of rice diversity in Central Asia. According to the results, 6 of 10, 2 of 7, and 0 of 6 of Afghan, Iranian, and Uzbek rice cultivars were scored as aromatic, respectively. Therefore, Afghan cultivars are a good source of aromatic rice germplasm for Central Asia. The expression between aromatic and non-aromatic, and aromatic and aromatic combinations has been evaluated. The observed segregation ratio of these crosses in the $F_2$ populations was tested by $x^2$ analysis against the expected ratio for a single gene. A segregation ratio of 3:1 between non-aromatic and aromatic combination has been detected, while segregation has not been detected between the aromatic and aromatic combinations. Also, parallel results were obtained from the tested aromatic rice cultivars. Thus, our results suggest that a single recessive gene controls aroma in all aromatic rice cultivars.

  • PDF