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Molecular and Cellular Probes

Libing Liu, Jianchang Wang, Yunyun Geng, Jinfeng Wang, Ruiwen Li, Ruihan Shi, Wanzhe Yuan
A visible and equipment-free recombinase polymerase amplification assay combined with a lateral flow strip (LFS RPA) was developed to detect canine parvovirus type 2 (CPV-2), which is the etiological agent of canine parvovirus disease. The CPV-2 LFS RPA assay was developed based on the VP2 gene and is performed in a closed fist using body heat for 15 min; the products are visible to the naked eye on the LFS within 5 min. The assay could detect CPV-2a, CPV-2b and CPV-2c, and there was no cross-reaction with the other viruses tested...
April 26, 2018: Molecular and Cellular Probes
Chunhe Wan, Longfei Cheng, Guanghua Fu, Cuiteng Chen, Rongchang Liu, Shaohua Shi, Hongmei Chen, Qiuling Fu, Yu Huang
Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-reactions with other common waterfowl viruses. The standard curve had a linear correlation of 0.997 and efficiency of 99% between the cycle threshold value and the logarithm of the plasmids copy number. The possible lowest detectable concentration was 35...
April 14, 2018: Molecular and Cellular Probes
Yueshuang Ke, Ke Wang, Hui Xu, Chenxin Wang, Jing Zhang, Xianlu Zeng, Ruoxi Wang, Istvan Boldogh, Xueqing Ba
Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor that catalyzes the poly (ADP-ribose) (PAR) onto a variety of target proteins, such as histones, DSB repair factors and PARP1 itself under consumption of NAD+ . Besides, PARP1 can affect a variety of proteins in noncovalent modification manner to carry out specific cellular functions. Here, we establishment a method to generate non-radiolabeled free PAR by PARG moderately cleaving PAR from autoPARylated PARP1, and utilize dot-blot assay to determine the interaction between free PAR and interested proteins...
April 5, 2018: Molecular and Cellular Probes
Wan Chunhe, Chen Cuiteng, Cheng Longfei, Fu Guanghua, Shi Shaohua, Fu Qiuling, Liu Rongchang, Chen Hongmei, Huang Yu
Pigeon torque teno virus (PTTV), a recently discovered circular DNA virus. Here, we developed a TaqMan-based real-time PCR for rapid and specific detection of PTTV infections with sensitivity up to 49.3 copies/μl. Positive signals can be observed by the assay in pigeon embryonated eggs, which indicted that PTTV can be transmitted vertically. Our findings play important implications for a better understanding the transmission of torque teno virus in pigeons.
April 3, 2018: Molecular and Cellular Probes
Yang Jie, Yi Chen, Zhi Yu, Hui Ding, Zhongfu Ma
Long-term exposure to traffic-related pollutants can lead to a variety of respiratory diseases, including inflammation, asthma, and lung cancer; however, the underlying biological mechanisms are not fully understood. We focused on the effects of exposure to different air pollutants on the expression of genes associated with inflammatory immune responses, allergic reactions and asthma, and lung cancer. In order to understand the cellular responses induced by exposure to different traffic-related pollutants, we performed PCR array to evaluate the mRNA expression of genes associated with inflammatory immune responses, allergic reactions and asthma, and lung cancer in the lungs of mice exposed to three different environments, including the laboratory (clean air), and polluted parking garages in Foshan and Guangzhou for four weeks...
April 2, 2018: Molecular and Cellular Probes
Waqar Islam
Efficient plant genome editing is dependent upon induction of double stranded DNA breaks (DSBs) through site specified nucleases. These DSBs initiate the process of DNA repair which can either base upon homologous recombination (HR) or non-homologous end jointing (NHEJ). Recently, CRISPR-Cas9 mechanism got highlighted as revolutionizing genetic tool due to its simpler frame work along with the broad range of adaptability and applications. So, in this review, we have tried to sum up the application of this biotechnological tool in plant genome editing...
April 2, 2018: Molecular and Cellular Probes
Junying Sun, Gali Bingga, Zhicheng Liu, Chunhong Zhang, Haiyan Shen, Pengju Guo, Jianfeng Zhang
Differentiation of classical strains and highly pathogenic strains of porcine reproductive and respiratory syndrome virus is crucial for effective vaccination programs and epidemiological studies. We used nested PCR and high resolution melting curve analysis with unlabeled probe to distinguish between the classical and the highly pathogenic strains of this virus. Two sets of primers and a 20 bp unlabeled probe were designed from the NSP3 gene. The unlabeled probe included two mutations specific for the classical and highly pathogenic strains of the virus...
March 30, 2018: Molecular and Cellular Probes
Murugan Rajalaxmi, Rajamohamed Beema Shafreen, Karuppiah Chithiraiselvi, Shunmugiah Karutha Pandian
This study aimed to determine the antibiofilm activity of seawater microbes against Vibrio cholerae (VCO1) through functional metagenomics approach. A metagenomic library was constructed from Palk Bay seawater and the library was screened to identify the biofilm inhibitory metaclone. Metaclone SWMC166 (harbouring ∼30 kb metagenomic insert) was found to exhibit antibiofilm activity against VCO1. The biofilm inhibitory potential of partially purified ethyl acetate extract of SWMC166 (EA166) was further evaluated through microscopic studies and biochemical assays...
March 21, 2018: Molecular and Cellular Probes
Wen Shi, Yuting Wang, Xuanyu Ren, Shuai Gao, Xiaojing Hua, Mengting Guo, Lijie Tang, Yigang Xu, Tong Ren, Yijing Li, Min Liu
Salmonid alphaviruses (SAVs), which include the etiological agents of salmon pancreas disease (PD) and sleeping disease (SD), are significant viral pathogens of European salmonid aquaculture, resulting in substantial economic losses to the salmonid-farming industry. Even though many countries including China have not reported the presence of SAV infections, these countries may be seriously threatened by these diseases as the salmon fish import trade increases. Thus, it is indeed necessary to develop efficient detection methods for the diagnosis and prevention of SAV infection...
March 16, 2018: Molecular and Cellular Probes
Maria Cryskely Agra Batinga, Julia Teresa Ribeiro de Lima, Fabio Gregori, Jaqueline Assumpção Diniz, Kerstin Muner, Trícia M F S Oliveira, Helena Lage Ferreira, Rodrigo Martins Soares, Lara Borges Keid
Canine brucellosis is caused by Brucella canis, a gram negative and facultative intracellular bacterium that is commonly associated with reproductive failures in dogs. The accurate diagnosis of the infection relies on the use of serological tests associated with blood culturing to guarantee sensitivity. The polymerase chain reaction (PCR) can replace the culturing procedure for the direct diagnosis of the infection because of its speed, high specificity and sensitivity values; however, it depends on some laboratory infrastructure to be conducted...
March 7, 2018: Molecular and Cellular Probes
Mohammad Ridhuan Mohd Ali, Amira Wahida Mohd Safee, Nor Hayati Ismail, Roslinda Abu Sapian, Hani Mat Hussin, Nabilah Ismail, Chan Yean Yean
BACKGROUND: Early diagnosis of leptospirosis is important for ensuring better clinical management and achieving better outcomes. Currently, serological assays suffer from inconsistent performance and are less useful for early diagnosis of leptospirosis. As an alternative, qPCR is more sensitive, specific and able to detect the presence of leptospiral DNA during the acute phase of the infection. Meanwhile, most molecular assays do not detect the non-pathogenic group of Leptospira, even though these groups may also infect humans, although less frequently and less severely...
April 2018: Molecular and Cellular Probes
Yaru Sun, Yuening Cheng, Peng Lin, Li Yi, Mingwei Tong, Zhigang Cao, Gaili Wang, Shuang Li, Shipeng Cheng, Wanzhe Yuan, Jianke Wang
Canine parvovirus (CPV) is an important pathogen in domestic dogs, and the original antigenic types CPV-2 and its variants, CPV-2a, 2b and 2c, are prevalent worldwide. A multiplex TaqMan real-time PCR method was developed for the detection and differentiation of four antigenic types of CPV. A set of primers and probes, CPV-305F/CPV-305R and CPV-2-305P (for CPV-2)/CPV-2a-305P (for CPV-2a, 2b and 2c), was able to differentiate CPV-2 and its variants (CPV-2a, 2b and 2c). Another set of primers and probes, CPV-426F/CPV-426R and CPV-2-426P (for CPV-2 and 2a)/CPV-2b-426P (for CPV-2b)/CPV-2c-426P (for CPV-2c), was able to differentiate CPV-2a (2), CPV-2b, and CPV-2c...
April 2018: Molecular and Cellular Probes
Hongmei Zhang, Kuiyu Wang, Shengjun Bu, Zhongyi Li, Chuanjing Ju, Jiayu Wan
Accurate and quantitative analysis of microRNA (miRNA) expression is critical for the diagnostics and theranostics of a disease. Herein, a proof-of-concept of a colorimetric horseradish peroxidase-mimicking DNAzyme (HRP-DNAzyme) biosensor for miRNA assay based on nuclease-assisted catalytic hairpin assembly (CHA) signal amplification was demonstrated. Duplex-specific nuclease (DSN) was employed to cleave the single-stranded DNA (ssDNA) chimeric probe (CP) on the magnetic bead (MB) surface via hybridization of the CP and target miRNA...
April 2018: Molecular and Cellular Probes
Xiaolong Zhou, Wentao Wang, Chengtao Du, Feifei Yan, Songbai Yang, Ke He, Han Wang, Ayong Zhao
OGG1 is the first enzyme in the base excision repair pathway (BER) responsible for repairing 8-oxoguanine DNA lesions. Recent studies found that OGG1 may also be involved in epigenetic regulation. In this study, we focused on the roles of OGG1 in histone modification. First, to study the effects of OGG1 on histone modification, the protein levels of symmetric dimethylation of histone H4 arginine-3 (H4R3me2s) were determined by western blot analysis following the knockdown or overexpression of OGG1. Second, the molecular mechanisms by which OGG1 regulates H4R3me2s were assessed by co-immunoprecipitation (CO-IP) assays in mouse embryonic fibroblast (MEF) wild-type (WT) and Ogg-/- cells...
April 2018: Molecular and Cellular Probes
Weidong Yao, Xin Wang, Kun Xiao
Berberine (BBR) is an isoquinnoline derivative alkaloid extracted from Rhizoma Coptidis that has the potential to protect myocardial tissues from ischemia/reperfusion (I/R) injuries. We attempted to evaluate the effect of BBR on the proliferation and apoptosis of a hypoxia/reoxygenation (H/R) cell model and to reveal the mechanism driving the improving function of BBR myocardial tissues. The H/R cell model was established using H9c2 rat cardiac myoblasts. The cell viability, apoptotic rates, and cell cycle distribution were measured with CCK-8 assay and flow cytometry...
April 2018: Molecular and Cellular Probes
Ronald J A Wanders
Peroxisomes catalyze a number of essential metabolic functions of which fatty acid alpha- and beta-oxidation, ether phospholipid biosynthesis, glyoxylate detoxification and bile acid synthesis are the most important. The key role of peroxisomes in humans is exemplified by the existence of a group of peroxisomal disorders, caused by mutations in > 30 different genes which code for proteins with a role in either peroxisome biogenesis or one of the metabolic pathways in peroxisomes. Technological advances in laboratory methods at the metabolite-, enzyme-, and molecular level have not only allowed the identification of new peroxisomal disorders but also new phenotypes associated with already identified genetic defects thus extending the clinical spectrum...
February 10, 2018: Molecular and Cellular Probes
Guiling Shi, Yijia Wang, Chunze Zhang, Zhenying Zhao, Xiuying Sun, Shiwu Zhang, Jinling Fan, Cunxia Zhou, Jihong Zhang, Huijuan Zhang, Jun Liu
BACKGROUND: Colorectal cancer (CRC) is a common cancer with high morbidity and mortality. However, its molecular mechanism is not clear, nor the genes related to CRC stages. METHODS: Gene expression data in CRC and healthy colorectal tissues were obtained from gene expression omnibus. Limma package was used to identify the differentially expressed genes (DEGs) between control and CRC (stage I, II, III, and IV), obtaining 4 DEG sets. VennPlex was utilized to find all DEGs and intersection DEGs...
February 2018: Molecular and Cellular Probes
Mingxun Li, Hailei Xia, Dan Chen, Dejun Ji, Tsunoda Kenji, Rui Li, Xiangxiang Liao, Yongjiang Mao, Wei Sun, Rongqing Geng, Zhangping Yang
To explore the genetic divergence and phylogeny of Chinese indigenous sheep breeds, in the current study, we analyzed the polymorphisms of 5 structural loci in ten sheep populations, including Sishui Fur, Sunite, Wurank, Bayinbuluke, Altay, Small-Tailed Han, Wadi, Tan, Tong and Hu sheep. The data were then compared with those from an additional 13 Asian and 4 European sheep populations acquired by the same experimental method. Based on the genetic distance and the results of a cluster analysis, we constructed the phylogenetic relationship of 27 populations...
February 2018: Molecular and Cellular Probes
Gnanavel Venkatesan, Mahesh Kumar Teli, Muthu Sankar, Amit Kumar, M Dashprakash, Sargam Arya, Aparna Madhavan, Muthannan Andavar Ramakrisnan, Awadh Bihari Pandey
The study is aimed to develop and evaluate a recombinant P32 protein based ELISA for sero-monitoring and sero-surveillance using known and random/suspected serum samples for capripox infections from sheep and goats. Truncated P32 gene of goatpox virus (with an ORF of 750 bp) was expressed in E. coli BL-21 CodonPlus (DE3)-RIPL cells using pET32a vector and characterized by SDS-PAGE analysis and confirmed by western blotting as 48 kDa polyhistidine-tagged fusion protein. The protein was purified under denaturing conditions using 8M urea and characterized by SDS-PAGE and immunoblotting...
February 2018: Molecular and Cellular Probes
Jing-Jing Li, Ya-Qin Chen, Liang-Liang Fan, Jie-Yuan Jin, Shuai Guo, Rong Xiang
Hypertriglyceridemia (HTG) plays an important role in the development and progression of atherosclerosis. It is inherited in an autosomal dominant pattern with a frequency of approximately 1:1,000,000 worldwide. Previous study has demonstrated that more than six genes underlie this disorder. In addition, copy number variants (CNVs) including disease-causing genes also play a crucial role in it. In this study, we have employed SNP-ARRAY chip technology to detect the pathogenic CNVs in a HTG patient who carried no meaningful mutations in HTG candidate genes...
February 2018: Molecular and Cellular Probes
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