• Title/Summary/Keyword: Molecular Detection

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Molecular characterization of Malaysian fowl adenovirus (FAdV) serotype 8b species E and pathogenicity of the virus in specific-pathogen-free chicken

  • Sabarudin, Nur Syazana;Tan, Sheau Wei;Phang, Yuen Fun;Omar, Abdul Rahman
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.42.1-42.16
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    • 2021
  • Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions: FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

Characteristics of the Infection of Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) in Compatible Wheat

  • Ren, Zhaoyu;Zhang, Wei;Wang, Mengke;Gao, Haifeng;Shen, Huimin;Wang, Chunping;Liu, Taiguo;Chen, Wanquan;Gao, Li
    • The Plant Pathology Journal
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    • v.37 no.5
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    • pp.437-445
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    • 2021
  • Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What's more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

Visualizing Live Chromatin Dynamics through CRISPR-Based Imaging Techniques

  • Chaudhary, Narendra;Im, Jae-Kyeong;Nho, Si-Hyeong;Kim, Hajin
    • Molecules and Cells
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    • v.44 no.9
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    • pp.627-636
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    • 2021
  • The three-dimensional organization of chromatin and its time-dependent changes greatly affect virtually every cellular function, especially DNA replication, genome maintenance, transcription regulation, and cell differentiation. Sequencing-based techniques such as ChIP-seq, ATAC-seq, and Hi-C provide abundant information on how genomic elements are coupled with regulatory proteins and functionally organized into hierarchical domains through their interactions. However, visualizing the time-dependent changes of such organization in individual cells remains challenging. Recent developments of CRISPR systems for site-specific fluorescent labeling of genomic loci have provided promising strategies for visualizing chromatin dynamics in live cells. However, there are several limiting factors, including background signals, off-target binding of CRISPR, and rapid photobleaching of the fluorophores, requiring a large number of target-bound CRISPR complexes to reliably distinguish the target-specific foci from the background. Various modifications have been engineered into the CRISPR system to enhance the signal-to-background ratio and signal longevity to detect target foci more reliably and efficiently, and to reduce the required target size. In this review, we comprehensively compare the performances of recently developed CRISPR designs for improved visualization of genomic loci in terms of the reliability of target detection, the ability to detect small repeat loci, and the allowed time of live tracking. Longer observation of genomic loci allows the detailed identification of the dynamic characteristics of chromatin. The diffusion properties of chromatin found in recent studies are reviewed, which provide suggestions for the underlying biological processes.

Imprinted Graphene-Starch Nanocomposite Matrix-Anchored EQCM Platform for Highly Selective Sensing of Epinephrine

  • Srivastava, Juhi;Kushwaha, Archana;Singh, Meenakshi
    • Nano
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    • v.13 no.11
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    • pp.1850131.1-1850131.19
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    • 2018
  • In this paper, an electrochemical sensor for epinephrine (EP), a neurotransmitter was developed by anchoring molecularly imprinted polymeric matrix (MIP) on the surface of gold-coated quartz crystal electrode of electrochemical quartz crystal microbalance (EQCM) using starch nanoparticles (Starch NP) - reduced graphene oxide (RGO) nanocomposite as polymeric format for the first time. Use of EP in therapeutic treatment requires proper dose and route of administration. Proper follow-up of neurological disorders and timely diagnosis of them has been found to depend on EP level. The MIP sensor was developed by electrodeposition of starch NP-RGO composite on EQCM electrode in presence of template EP. As the imprinted sites are located on the surface, high specific surface area enables good accessibility and high binding affinity to template molecule. Differential pulse voltammetry (DPV) and piezoelectrogravimmetry were used for monitoring binding/release, rebinding of template to imprinted cavities. MIP-coated EQCM electrode were characterized by contact angle measurements, AFM images, piezoelectric responses including viscoelasticity of imprinted films, and other voltammetric measurements including direct (DPV) and indirect (using a redox probe) measurements. Selectivity was assessed by imprinting factor (IF) as high as 3.26 (DPV) and 3.88 (EQCM). Sensor was rigorously checked for selectivity in presence of other structurally close analogues, real matrix (blood plasma), reproducibility, repeatability, etc. Under optimized conditions, the EQCM-MIP sensor showed linear dynamic ranges ($1-10{\mu}M$). The limit of detection 40 ppb (DPV) and 290 ppb (EQCM) was achieved without any cross reactivity and matrix effect indicating high sensitivity and selectivity for EP. Hence, an eco-friendly MIP-sensor with high sensitivity and good selectivity was fabricated which could be applied in "real" matrices in a facile manner.

Occurrence and control of N-nitrosodimethylamine in water engineering systems

  • Bian, Yongning;Wang, Chuang;Zhu, Guocheng;Ren, Bozhi;Zhang, Peng;Hursthouse, Andrew S.
    • Environmental Engineering Research
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    • v.24 no.1
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    • pp.1-16
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    • 2019
  • N-nitrosodimethylamine (NDMA) is a typical nitrogen disinfection by-product, which has posed a potential threat to human health during drinking water disinfection. Because of the well-known effects of mutagenesis, carcinogenesis and teratogenesis, the high detection rate in water engineering systems (such as coagulation, membrane filtration and biological systems), and difficulty to remove, it has received wide concern in the field of water engineering systems. The NDMA is a low molecular weight hydrophilic organic substance, which is difficult to remove. Also, the mechanism for NDMA formation is also recognized to be complex, and many steps still needed to be further evaluated. Therefore, the mechanistic knowledge on NDMA formation potential and their removal processes is of particularly interest. Few papers summarize the occurrence and control of NDMA in water engineering systems. It is for this reason that the content of this paper is particularly important for us to understand and control the amount of NDMA thus reducing the threat of disinfection by-products to drinking water. Four parts including the mechanisms for the NDMA formation potential, the factors affecting the NDMA formation potential, the technologies for removal of NDMA are summarized. Finally, some definite suggestions are given.

Molecular and Morphological Evidence of Hepatotoxicity after Silver Nanoparticle Exposure: A Systematic Review, In Silico, and Ultrastructure Investigation

  • Sooklert, Kanidta;Wongjarupong, Asarn;Cherdchom, Sarocha;Wongjarupong, Nicha;Jindatip, Depicha;Phungnoi, Yupa;Rojanathanes, Rojrit;Sereemaspun, Amornpun
    • Toxicological Research
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    • v.35 no.3
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    • pp.257-270
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    • 2019
  • Silver nanoparticles (AgNPs) have been widely used in a variety of applications in innovative development; consequently, people are more exposed to this particle. Growing concern about toxicity from AgNP exposure has attracted greater attention, while questions about nanosilver-responsive genes and consequences for human health remain unanswered. By considering early detection and prevention of nanotoxicology at the genetic level, this study aimed to identify 1) changes in gene expression levels that could be potential indicators for AgNP toxicity and 2) morphological phenotypes correlating to toxicity of HepG2 cells. To detect possible nanosilver-responsive genes in xenogenic targeted organs, a comprehensive systematic literature review of changes in gene expression in HepG2 cells after AgNP exposure and in silico method, connection up- and down-regulation expression analysis of microarrays (CU-DREAM), were performed. In addition, cells were extracted and processed for transmission electron microscopy to examine ultrastructural alterations. From the Gene Expression Omnibus (GEO) Series database, we selected genes that were up- and down-regulated in AgNPs, but not up- and down-regulated in silver ion exposed cells, as nanosilver-responsive genes. HepG2 cells in the AgNP-treated group showed distinct ultrastructural alterations. Our results suggested potential representative gene data after AgNPs exposure provide insight into assessment and prediction of toxicity from nanosilver exposure.

Malaria Prevalence in a Low Transmission Area, Jazan District of Southwestern Saudi Arabia

  • Hawash, Yousry;Ismail, Khadiga;Alsharif, Khalaf;Alsanie, Walaa
    • Parasites, Hosts and Diseases
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    • v.57 no.3
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    • pp.233-242
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    • 2019
  • Detailed description of malaria in low transmission areas is crucial for elimination. The current study aimed to provide a comprehensive description for malaria transmission in Jazan, a low transmission district, southwestern Saudi Arabia. Patients at a tertiary care hospital were recruited in our study between August 2016 and September 2018. Malaria diagnosis was performed through a species-specific nested polymerase chain reaction (nested PCR), microscopy and Paramax-$3^{TM}$ rapid detection test (RDT). Malaria was detected in 30 patients by the PCR, with point prevalence of 10.9%. Of these malaria infections, 80% was imported, 26.6% was asymptomatic and 23.3% was sub-microscopic. Malaria was reported throughout the year, with February/March and September/October peaks. Infection was significantly more in males than in females (P=0.01). Likewise, infections were detected more in febrile than in non-febrile patients (P=0.01). Adult aged 15-24 years, fever and travel were identified as high-risk factors. Malaria was primarily attributed to Plasmodium falciparum mono-infections, followed by P. vivax mono-infections and lastly to falciparum/vivax mixed infections accounting 76.6%, 16.6%, and 6.6% of PCR-confirmed malaria cases, respectively. The nested PCR was superior to the smear microscopy (sensitivity 76.6%; specificity 100%) and the RDT (sensitivity 83.3%, specificity 94.2%). The overall percent agreement between microscopy and the RDT was 92.7% (kappa=0.63). High proportion of imported malaria including sub-microscopic and sub-patent cases were described. We suggest that incorporation of molecular tool into the conventional malaria diagnosis is beneficial in Jazan district.

Research Trend of Biochip Sensors for Biomarkers Specific to Diagnostics of Lung Cancer Diseases (폐암 질환 진단에 활용 가능한 바이오마커 검출용 바이오칩 센서 연구 동향)

  • Lee, Sang Hyuk;Goh, Eunseo;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.29 no.6
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    • pp.645-651
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    • 2018
  • Lung cancer has the highest death rate of any cancer diseases in Koreans. However, patients often feel difficult to recognize their disease before facing the terminal diagnosis due to the absence of any significant symptoms. Furthermore, the clear detection of an early cancer stage is usually obscure with existing diagnostic methods. For this reason, extensive research efforts have been made on introducing a wide range of biochemical diagnostic tools for the molecular level analysis of biological fluids for lung cancer diagnoses. A chip-based biosensor, one type of the analytical devices, can be a great potential for the diagnosis, which can be used without any further expensive analytical equipments nor skilled analysts. In this mini review, we highlight recent research trends on searching biomarker candidates and bio-chip sensors for lung cancer diagnosis in addition to discussing their future aspects.

Analysis of Scutellaria baicaleinsis Georgi (Scutellariae Radix) by LC-DAD and LC-ESI/MS

  • Yu, Youngbob;Choi, Pil-Son;Koo, Sungtae;Chang, Suhwan
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.652-659
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    • 2018
  • In this study, baicalin, as a marker substance of Scutellariae Radix, was quantitatively analyzed by a high performance liquid chromatography-photodiode array detector (HPLC-DAD). We identified wogonoside, baicalein, and wogonin in the Scutellariae Radix by a high performance liquid chromatography-electrospray ionization-mass spectrometer (HPLC-ESI-MS). The baicalin was separated on a Xterra C18 column ($5{\mu}m$, $4.6{\times}250mm$) using mobile phase consisting of 38% acetonitrile in 0.68% phosphoric acid. The baicalin spectrum in the Scutellariae Radix extracts was coincided by comparing with UV-visible spectrum (200-550 nm) of baicalin standard in the library. The amount of baicalin in Scutellariae Radix was 10.46%, which is higher than KFDA's guideline. The marker substances of Scutellariae Radix showed a strong base peak $[M]^+$ in the positive detection mode following as; baicalin (m/z; $271[MH^+-sugar]^+$, $447[M+H]^+$), wogonoside (m/z; $285[MH^+-sugar]^+$, $461[M+H]^+$), baicalein (m/z; $271\;[M+H]^+$), wogonin (m/z; $285[M+H]^+$). These results are consistent with the fragment pattern and molecular weight of standard components from literature.

Identification of Toxic Chemicals Using Polypyrrole-Cyclodextrin Hybrids (폴리피롤-사이클로덱스트린 혼성체를 이용한 유해화합물질의 검출)

  • Bae, Joonwon
    • Applied Chemistry for Engineering
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    • v.30 no.2
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    • pp.186-189
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    • 2019
  • Polypyrrole is a typical electrical conducting polymer, which has an excellent charge transport property. Cyclodextrins are a group of toxic-free and cyclic oligosaccharide molecules, capable of capturing low molecular weight chemicals. Considering these advantages, hybrid materials of polypyrrole and cyclodextrin can be used to detect hazardous compounds. Cyclodextrin molecules can accommodate toxic chemicals by the formation of host-guest complexes and generate electric signals, which are effectively delivered by polypyrrole backbone. In this study, the polypyrrole/cyclodextrin hybrid material was prepared using a facile wet method and included into a hydrogel. Subsequently, it was applied to a simple sensor system with a gold-patterned electrode for the detection of potentially hazardous material, methyl paraben. Compared with pristine polypyrrole, it was found that the polypyrrole/cyclodextrin hybrid showed an improved performance. This study can be an example of using environmentally benign conducting polymer/cyclodextrin hybrids as sensing media.