• Title/Summary/Keyword: molecular monitoring

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Microchips and their Significance in Isolation of Circulating Tumor Cells and Monitoring of Cancers

  • Sahmani, Mehdi;Vatanmakanian, Mousa;Goudarzi, Mehdi;Mobarra, Naser;Azad, Mehdi
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.879-894
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    • 2016
  • In micro-fluid systems, fluids are injected into extremely narrow polymer channels in small amounts such as micro-, nano-, or pico-liter scales. These channels themselves are embedded on tiny chips. Various specialized structures in the chips including pumps, valves, and channels allow the chips to accept different types of fluids to be entered the channel and along with flowing through the channels, exert their effects in the framework of different reactions. The chips are generally crystal, silicon, or elastomer in texture. These highly organized structures are equipped with discharging channels through which products as well as wastes of the reactions are secreted out. A particular advantage regarding the use of fluids in micro-scales over macro-scales lies in the fact that these fluids are much better processed in the chips when they applied as micro-scales. When the laboratory is miniaturized as a microchip and solutions are injected on a micro-scale, this combination makes a specialized construction referred to as "lab-on-chip". Taken together, micro-fluids are among the novel technologies which further than declining the costs; enhancing the test repeatability, sensitivity, accuracy, and speed; are emerged as widespread technology in laboratory diagnosis. They can be utilized for monitoring a wide spectrum of biological disorders including different types of cancers. When these microchips are used for cancer monitoring, circulatory tumor cells play a fundamental role.

Application of Recent DNA/RNA-based Techniques in Rumen Ecology

  • McSweeney, C.S.;Denman, S.E.;Wright, A.-D.G.;Yu, Z.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.283-294
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    • 2007
  • Conventional culture-based methods of enumerating rumen microorganisms (bacteria, archaea, protozoa, and fungi) are being rapidly replaced by nucleic acid-based techniques which can be used to characterise complex microbial communities without incubation. The foundation of these techniques is 16S/18S rDNA sequence analysis which has provided a phylogenetically based classification scheme for enumeration and identification of microbial community members. While these analyses are very informative for determining the composition of the microbial community and monitoring changes in population size, they can only infer function based on these observations. The next step in functional analysis of the ecosystem is to measure how specific and, or, predominant members of the ecosystem are operating and interacting with other groups. It is also apparent that techniques which optimise the analysis of complex microbial communities rather than the detection of single organisms will need to address the issues of high throughput analysis using many primers/probes in a single sample. Nearly all the molecular ecological techniques are dependant upon the efficient extraction of high quality DNA/RNA representing the diversity of ruminal microbial communities. Recent reviews and technical manuals written on the subject of molecular microbial ecology of animals provide a broad perspective of the variety of techniques available and their potential application in the field of animal science which is beyond the scope of this treatise. This paper will focus on nucleic acid based molecular methods which have recently been developed for studying major functional groups (cellulolytic bacteria, protozoa, fungi and methanogens) of microorganisms that are important in nutritional studies, as well as, novel methods for studying microbial diversity and function from a genomics perspective.

Development of Species-specific Molecular Marker as a Tool for Discrimination between Crucian Carp Gengorobuna (Carassius cuvieri) Introduced from Japan and Korean Native One (C. auratus) (국내 자연산 붕어와 일본에서 도입된 떡붕어를 구분하기 위한 종특이적 분자마커 개발)

  • Song, Kyo-Hong;Jung, Jong-Woo;Koo, Hye-Young;Kim, Won
    • Korean Journal of Ecology and Environment
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    • v.40 no.1
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    • pp.143-148
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    • 2007
  • The introduced exotic species has often caused severe problems to the native ecosystem. One of such species is the freshwater fish gengorobuna (Carassius cuvieri) introduced from Japan. The first step to assess harmful effects of this species on the Korean freshwater ecosystem is to discriminate it from the most similar native crucian carp (Carassius auratus). Because traditional morphological identification often gives unreliable results due to their highly similar phenotype, a new more efficient method is needed. For this purpose, molecular markers produced by the efficient one-step PCR method using three primers (DDF, DDR and DDR1) were developed and tested in the present study. This molecular marker will play an important role in monitoring fish community of Korean freshwater ecosystem.

Relationship between Preoperative Serum CA15-3 and CEA Levels and Clinicopathological Parameters in Breast Cancer

  • Moazzezy, Neda;Farahany, Tahereh Zarnoosheh;Oloomi, Mana;Bouzari, Saeid
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1685-1688
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    • 2014
  • Background: CEA and CA 15.3 serum tumor markers are currently used in clinical practice for monitoring therapy. The aim of this study was to evaluate serum level of these markers among healthy females and invasive breast carcinoma (IBC) patients and to determine any relationships with clinicopathological factors. Materials and Methods: 60 Iranian females were enrolled in this study, 30 healthy and 30 diagnosed with breast cancer who had not received any preoperative chemotherapy or hormone therapy. Enzyme linked immunosorbent assays were used for the quantitative determination of the cancer associated antigens, CEA and MUC1 (CA15-3). Results: The serological levels of CEA and CA15-3 ($5.0033{\pm}0.49{\mu}g/L$ and $178.1667{\pm}15.11$ U/ml) in the breast cancer patients were significantly higher (p=0.00) than the serum levels of normal controls ($1.1237{\pm}0.11{\mu}g/L$ and $21.13{\pm}3.058$ U/ml). Regarding the CEA marker, a significant correlation with grade of tumor was shown. Furthermore, there was a low correlation between CA15-3 and CEA marker with correlation coefficient r=0.08. Conclusions: Collectively, markedly high levels of CEA and CA15-3 were found in our patients, pointing to their use as additional tools after clinical diagnosis.

Plasmodium vivax Drug Resistance Genes; Pvmdr1 and Pvcrt-o Polymorphisms in Relation to Chloroquine Sensitivity from a Malaria Endemic Area of Thailand

  • Rungsihirunrat, Kanchana;Muhamad, Poonuch;Chaijaroenkul, Wanna;Kuesap, Jiraporn;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • v.53 no.1
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    • pp.43-49
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    • 2015
  • The aim of the study was to explore the possible molecular markers of chloroquine resistance in Plasmodium vivax isolates in Thailand. A total of 30 P. vivax isolates were collected from a malaria endemic area along the Thai-Myanmar border in Mae Sot district of Thailand. Dried blood spot samples were collected for analysis of Pvmdr1 and Pvcrt-o polymorphisms. Blood samples ($100{\mu}l$) were collected by finger-prick for in vitro chloroquine susceptibility testing by schizont maturation inhibition assay. Based on the cut-off $IC_{50}$ of 100 nM, 19 (63.3%) isolates were classified as chloroquine resistant P. vivax isolates. Seven non-synonymous mutations and 2 synonymous were identified in Pvmdr1 gene. Y976F and F1076L mutations were detected in 7 (23.3%) and 16 isolates (53.3%), respectively. Analysis of Pvcrt-o gene revealed that all isolates were wild-type. Our results suggest that chloroquine resistance gene is now spreading in this area. Monitoring of chloroquine resistant molecular markers provide a useful tool for future control of P. vivax malaria.

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.

Trend of In Silico Prediction Research Using Adverse Outcome Pathway (독성발현경로(Adverse Outcome Pathway)를 활용한 In Silico 예측기술 연구동향 분석)

  • Sujin Lee;Jongseo Park;Sunmi Kim;Myungwon Seo
    • Journal of Environmental Health Sciences
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    • v.50 no.2
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    • pp.113-124
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    • 2024
  • Background: The increasing need to minimize animal testing has sparked interest in alternative methods with more humane, cost-effective, and time-saving attributes. In particular, in silico-based computational toxicology is gaining prominence. Adverse outcome pathway (AOP) is a biological map depicting toxicological mechanisms, composed of molecular initiating events (MIEs), key events (KEs), and adverse outcomes (AOs). To understand toxicological mechanisms, predictive models are essential for AOP components in computational toxicology, including molecular structures. Objectives: This study reviewed the literature and investigated previous research cases related to AOP and in silico methodologies. We describe the results obtained from the analysis, including predictive techniques and approaches that can be used for future in silico-based alternative methods to animal testing using AOP. Methods: We analyzed in silico methods and databases used in the literature to identify trends in research on in silico prediction models. Results: We reviewed 26 studies related to AOP and in silico methodologies. The ToxCast/Tox21 database was commonly used for toxicity studies, and MIE was the most frequently used predictive factor among the AOP components. Machine learning was most widely used among prediction techniques, and various in silico methods, such as deep learning, molecular docking, and molecular dynamics, were also utilized. Conclusions: We analyzed the current research trends regarding in silico-based alternative methods for animal testing using AOPs. Developing predictive techniques that reflect toxicological mechanisms will be essential to replace animal testing with in silico methods. In the future, since the applicability of various predictive techniques is increasing, it will be necessary to continue monitoring the trend of predictive techniques and in silico-based approaches.

Comprehensive investigations of key mitochondrial metabolic changes in senescent human fibroblasts

  • Ghneim, Hazem K.;Alfhili, Mohammad A.;Alharbi, Sami O.;Alhusayni, Shady M.;Abudawood, Manal;Aljaser, Feda S.;Al-Sheikh, Yazeed A.
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.263-275
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    • 2022
  • There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H2O2), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H2O2, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H2O2, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD+/NADH and NADP+/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

A Study on Match and Mismatch DNA Hybridization properties Using DNA Hybridization Detection Sensor (DNA Hybridization 검출 센서를 이용한 매치 및 미스매치 DNA hybridization 특성 연구)

  • Kim, Do-Kyun;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.89-91
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    • 2003
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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Affecting Factors on the Variation of Atmospheric Concentration of Polycyclic Aromatic Hydrocarbons in Central London

  • Baek, Sung-Ok;Roger Perry
    • Journal of Korean Society for Atmospheric Environment
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    • v.10 no.E
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    • pp.343-356
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    • 1994
  • In this study, a statistical investigation was carried out for the evaluation of any relationship between polycyclic aromatic hydrocarbons (PAHss) associated with ambient aerosols and other air quality parameters under varying meteorological conditions. Daily measurements for PAHs and air quality/meteorological parameters were selected from a data-base constructed by a comprehensive air monitoring in London during 1985-1987. Correlation coefficients were calculated to examine any significant relationship between the PAHs and other individual variables. Statistical analysis was further Performed for the air quality/meteorological data set using a principal component analysis to derive important factors inherent in the interactions among the variables. A total of six components were identified, representing vehicle emission, photochemical activity/volatilization, space heating, atmospheric humidity, atmospheric stability, and wet deposition. It was found from a stepwise multiple regression analysis that the vehicle emission component is overall the most important factor contributing to the variability of PAHs concentrations at the monitoring site. The photochemical activity/volatilzation component appeared to be also an important factor particularly for the lower molecular weight PAHs. In general, the space heating component was found to be next important factor, while the contributions of other three components to the variance of each PAHs did not appear to be as much important as the first three components in most cases. However, a consistency for these components in their negative correlations with PAHs data was found, indicating their roles in the depletion of PAHs concentrations in the urban atmosphere.

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