• 제목/요약/키워드: molecular contamination

검색결과 147건 처리시간 0.025초

Utility of taxon-specific molecular markers for the species identification of herbarium specimens: an example from Desmarestia japonica (Phaeophyceae, Desmarestiales) in Korea

  • Lee, Sang-Rae;Lee, Eun-Young
    • Fisheries and Aquatic Sciences
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    • 제21권3호
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    • pp.8.1-8.6
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    • 2018
  • Desmarestia japonica (Phaeophyceae, Desmarestiales) was recently established from the Japanese ligulate Desmarestia and is morphologically similar to D. ligulata. This species has been reported only from Japan. However, the taxonomic reports based on additional regional distributions are needed to clarify this taxonomic entity and its species boundaries. Because Desmarestia species have restricted distributions in Korea, we reexamined herbarium specimens of D. ligulata deposited at the National Institute of Biological Resources (South Korea). To improve the amplification efficiency of the polymerase chain reaction and avoid contamination by the DNA of other organisms, we developed taxon-specific molecular markers suitable for DNA barcoding of Desmarestia species. Nuclear ribosomal small subunit RNA (18S rDNA) and mitochondrial cytochrome c oxidase 1 (cox1) regions were selected as target DNA. As a result, both were successfully isolated from herbarium specimens of D. japonica acquired over 10 years. These molecular markers provide useful genetic information for herbarium specimens for which conventional molecular analysis is challenging.

False-Positive Mycobacterium tuberculosis Detection: Ways to Prevent Cross-Contamination

  • Asgharzadeh, Mohammad;Ozma, Mahdi Asghari;Rashedi, Jalil;Poor, Behroz Mahdavi;Agharzadeh, Vahid;Vegari, Ali;Shokouhi, Behrooz;Ganbarov, Khudaverdi;Ghalehlou, Nima Najafi;Leylabadlo, Hamed Ebrahmzadeh;Kafil, Hossein Samadi
    • Tuberculosis and Respiratory Diseases
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    • 제83권3호
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    • pp.211-217
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    • 2020
  • The gold standard method for diagnosis of tuberculosis is the isolation of Mycobacterium tuberculosis through culture, but there is a probability of cross-contamination in simultaneous cultures of samples causing false-positives. This can result in delayed treatment of the underlying disease and drug side effects. In this paper, we reviewed studies on false-positive cultures of M. tuberculosis. Rate of occurrence, effective factors, and extent of false-positives were analyzed. Ways to identify and reduce the false-positives and management of them are critical for all laboratories. In most cases, false-positive is occurring in cases with only one positive culture but negative direct smear. The three most crucial factors in this regard are inappropriate technician function, contamination of reagents, and aerosol production. Thus, to reduce false-positives, good laboratory practice, as well as use of whole-genome sequencing or genotyping of all positive culture samples with a robust, extra pure method and rapid response, are essential for minimizing the rate of false-positives. Indeed, molecular approaches and epidemiological surveillance can provide a valuable tool besides culture to identify possible false positives.

상온 노화 후 고무가황물에서 왁스의 이동과 증발에 미치는 왁스의 분자량 분포 (Influence of Wax Molecular Weights on Wax Migration and Evaporation of Rubber Vulcanizates at Room Temperature)

  • 임송희;최성신
    • Elastomers and Composites
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    • 제44권4호
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    • pp.397-400
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    • 2009
  • 오존은 상온에서 폴리머 이중 결합의 산화작용을 활발하게 하여 표면 갈라짐을 유발시킨다. 왁스는 고무 표면으로 이동하여 물리적 방어막을 형성하여 오존의 작용을 방지한다. 비보강 NR, SBR, BR 가황물을 상온에서 6개월간 노화 시켜 노화 전후의 왁스의 분자량 분포 변화를 조사하였다. 상온 조건에서 고무 가황물에 있는 왁스의 이동과 증발 거동은 노화방지제로서의 왁스의 역할과 고무 제품의 외관 오염을 이해하는데 도움을 준다. NR 시험편에서 알칸의 상대 세기 분포는 노화 후 고분자량쪽으로 이동하였으나 SBR 시험편의 경우에는 노화 전후의 분자량 분포 모양에는 큰 차이가 없었다.

MOLECULAR BIOMARKER OF CADMIUM EXPOSURE IN FRESHWATER FISH: SENSITIVITY AND SPECIFICITY

  • Park, Kwangsik;Heekyung Bae;Nam, Seong-Sook;Kim, Enkyoung
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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    • pp.174-174
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    • 2002
  • Metallothioneins (MTs) are known to be induced by heavy metals in various organs of different species and represent a potential biomarker of aquatic contamination by heavy metals. In this work, cloning and sequencing of a metallothionein gene in crucian carp (Carassius auratus) was done and sensitivities and specificities of the gene expressions were compared.(omitted)

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Rapid Analysis of Melamine Content in Powdered and Liquid Milk Using Fourier Transform Infrared Spectroscopy

  • Wang, Jun;Jun, Soo-Jin;Li, Qing X.
    • Food Science and Biotechnology
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    • 제18권5호
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    • pp.1199-1203
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    • 2009
  • Melamine is a chemical intermediate to manufacture amino resins and plastics, which cannot be used as food additive since it can cause kidney stones. A qualitative determination method of melamine residue in powdered and liquid milk was developed using Fourier transform infrared (FTIR) spectroscopic technique. The calibration sets consisted of 21 standard melamine solutions, in which 1% trichloroacetic acid aqueous solution and acetonitrile (3:1, v/v) were used as solvent. The model was validated using 10 standard melamine solutions which were unused to build up the calibration set. Infrared (IR) absorbance peaks specific to almost all chemical groups in melamine molecule were shown in the spectral range between 1,100 and 1,800/cm. Combined partial least squares (PLS)-$2^{nd}$ derivative calibration model coupled with mean centering (MC) mathematical enhancement showed the highest correlation coefficients ($R^2$>0.99). In brief, the FTIR technique can be used for quantitative analysis of melamine in milk samples.

출토 인골의 분자유전학 및 조직학적 분석연구 (Molecular Genetic and Histological Analysis for the Excavated Human Bone)

  • 지상현;박지희;서민석;홍종욱;정용재
    • 보존과학연구
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    • 통권28호
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    • pp.75-90
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    • 2007
  • The ancient biomolecular remains are the potential source for paleobiology and paleoanthropology. Especially, ancient human specimens such as bone, teeth, and hair are powerful materials to identify historical origin and migration of ancestor population from the past. However, most excavated human specimens in archaeological sites have commonly problems as natural damage and exogenous contamination. We carried out histological and molecular analyses of excavated bone from the historic sites in South Korea from the recently discovered in tumulus of Seochun and Naju. Biological deterioration of bone was observed anatomically by optical and scanning electron microscope (SEM). We extracted degraded DNA, and amplified hyper variable region (HVR) of mitochondrial DNA (mtDNA) and amelogenin of nucleus DNA. This study applied and examined the relationships between histological preservation and DNA survival in excavated bone.

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대장균의 회분식 발효에 의해 생산된 덱스트란 결합 파아지를 활용한 설탕 제조공정 오염 검출 (Detection of Sugar Process Contamination Using Dextran Binding hages Produced by Batch Fermentation of Escherichia coli)

  • 김두운
    • 한국식품저장유통학회지
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    • 제15권5호
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    • pp.617-621
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    • 2008
  • Sequential passes through $Sephadex^{TM}$ columns were used to select phages that displays ligands for dextran ($\alpha$-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the $Sephadex^{TM}$ in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave $72\;{\pm}\;10$ normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, $2\;{\times}\;10^6) and significantly lower color ($30\;{\pm}\;6$ N.I.) against low molecular size dextran (T10, $10^4$). The presence of an Fab insert in each of these phages was confirmed using a $\beta-galactosidase linked assay and polymerase chain reaction.

Gene expression profiling after ochratoxin A treatment in small intestinal epithelial cells from pigs

  • Jung Woong, Yoon;Sang In, Lee
    • Journal of Animal Science and Technology
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    • 제64권5호
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    • pp.842-853
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    • 2022
  • Ochratoxin A (OTA) is a well-known mycotoxin that causes disease through the ingestion of contaminated food or feed, for example, in the porcine industry. The intestinal epithelium acts as the first barrier against food contamination. We conducted a study on the exposure of the porcine intestinal epithelium to OTA. We used the intestinal porcine epithelial cell line IPEC-J2 as an in vitro model to evaluate the altered molecular mechanisms following OTA exposure. Gene expression profiling revealed that OTA upregulated 782 genes and downregulated 896, totalling 1678 differentially expressed genes. Furthermore, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting confirmed that OTA damages the tight junction protein ZO-1. Moreover, OTA activated the expression of inflammatory genes (IL-6, IL-8, IL-10, NF-kB, TLR4, and TNF-α). In summary, this study confirmed that OTA alters various molecular mechanisms and has several adverse effects on IPEC-J2 cells.

Sialoglycoproteins of Mammalian Erythrocyte Membranes: A Comparative Study

  • Sharma, Savita;Gokhale, Sadashiv M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권12호
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    • pp.1666-1673
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    • 2011
  • The presence of sialoglycoproteins (SGPs) in the membranes from goat (Capra aegagrus hircus), buffalo (Bubalus bubalis bubalis) and pig (Sus scrofa domestica) erythrocytes was investigated by partial purification with a chloroform-methanol extraction method followed by Sodium dodecyl sulphate - Polyacrylamide gel electrophoresis in comparison to human (Homo sapiens) erythrocytes. The results show that mammalian erythrocytes possess clear differences in the SGPs numbers and molecular weights although all animals studied in this experiment are from the same class i.e. mammalia. The SGPs number in human, goat, buffalo and pig are four (PAS-1 to PAS-4), ten (PAS-GI to PAS-GX), seven (PAS-BI to PAS-BVII) and four (PAS-PI to PAS-IV) respectively as indicated by staining the polyacrylamide gel with sialoglycoprotein-specific Periodic acid-Schiff's (PAS) stain. The new SGPs could be observed only after the partial purification of membrane fractions named as PAS-HI with molecular weight (Mr) 190 kDa and PAS-HII 150 kDa in human, PAS-BIA in buffalo and PAS-PIA and PAS-PIVA in pig. The gels were also stained with Coomassie brilliant blue (CBB) and Silver stain to check the contamination of other membrane proteins in the purified fractions. The quantitative distribution of SGPs was also determined by densitometry. Present study indicates that there are some basic differences in mammalian erythrocyte membrane SGPs, especially with respect to their number and molecular weights indicating major structural variations.

Gene Expression Profile in Iprobenfos Exposed Medaka Fish by Microarray Analysis

  • Woo, Seon-Ock;Son, Sung-Hee;Ryu, Jae-Chun;Yum, Seung-Shic
    • Molecular & Cellular Toxicology
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    • 제4권2호
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    • pp.132-137
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    • 2008
  • Differential gene expression profiling was carried out in the hepatic tissue of medaka fish, Oryzias latipes, after exposure to an organophosphorus pesticide (OPP), Iprobenfos (IBP), a widely used pesticide in agri- and fish-culture, using a medaka cDNA micro array. Twenty six kinds of differentially expressed candidate genes, with 15 and 11 induced and repressed in their gene expressions, respectively, were associated with cytoskeleton (3.8%), development (7.7%), immune (7.7%), metabolism (30.8%), nucleic acid/protein binding (42.3%) and reproduction (7.7%). Of these genes, changes at the transcription level of five were re-evaluated by real-time quantitative PCR (qRT-PCR). Considering the known function of authentic genes, the effects of IBP on the biological activity and pathological aspects in medaka fish were discussed. The identified genes could be used as molecular biomarkers for biological responses to OPPs contamination in an aquatic environment.