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https://www.readbyqxmd.com/read/29776628/molecular-pathology-techniques-advances-in-2018
#1
REVIEW
Mark J Bluth, Martin H Bluth
Molecular pathology techniques continue to evolve. Although polymerase chain reaction (PCR) remains the cornerstone methodology for nucleic acid amplification, improvements in nucleic acid detection methodologies (i.e. PCR) have increased the detection sensitivity by using fluorescent and bead based array technologies. Single base pair lesions can be detected via sequencing and related techniques to discern point mutations in disease pathogenesis. Novel technologies, such as high- resolution melting analysis, provide fast high throughput post PCR analysis of genetic mutations or variance in nucleic acid sequences...
June 2018: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29775853/an-ultrasensitive-electrochemiluminescence-biosensor-for-detection-of-microrna-by-in-situ-electrochemically-generated-copper-nanoclusters-as-luminophore-and-tio-2-as-coreaction-accelerator
#2
Hongxia Liao, Ying Zhou, Yaqin Chai, Ruo Yuan
Herein, we constructed an ultrasensitive electrochemiluminescence (ECL) biosensor for detecting microRNA-21 (miR-21) based on in-situ generation of copper nanoclusters (Cu NCs) as luminophore and titanium dioxide (TiO2 ) as coreaction accelerator. First, numerous AT-rich double-stranded DNA (dsDNA) was produced from the conversion of a small amount of target miR-21 via the combination of exonuclease III (Exo III)-assisted amplification and hybridization chain reaction (HCR), which could reduce the aggregation-caused self-etching effect of Cu NCs and improve the emitting of Cu NCs...
May 8, 2018: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29774905/dual-emissions-and-thermochromic-luminescences-of-isomorphic-chiral-twofold-interpenetrated-3-d-nets-built-from-i-1-o-2-type-hybrid-inorganic-organic-frameworks-of-nh-2-ch-3-2-3-pb-2-x-3-bdc-2-x-br-i
#3
Shao-Xian Liu, Guo-Jun Yuan, Lu Zhai, Lifeng Wang, Hong-Bin Luo, Xiao-Ming Ren
Two pairs of MOF-based hybrid enantiomorphs, [NH2(CH3)2]3[Pb2X3(BDC)2] (H2BDC = 1,4-benzenedicarboxylic acid, X = Br or I), have been synthesized using the solvothermal reaction and then manually separated, which are labeled as 1a/1b (X = Br) and 2a/2b (X = I). The isomorphic 1a and 2a crystallize in tetragonal space group P43212, and the isomorphic 1b and 2b in the enantiomorphic space group P41212. Twofold interpenetrated three-dimensional (3-D) networks were built from two sets of equivalent I1O2 type hybrid inorganic-organic frameworks in 1a/1b and 2a/2b...
May 18, 2018: Dalton Transactions: An International Journal of Inorganic Chemistry
https://www.readbyqxmd.com/read/29774065/catalytic-hairpin-assembly-gel-assay-for-multiple-and-sensitive-microrna-detection
#4
Wenhao Dai, Jing Zhang, Xiangdan Meng, Jie He, Kai Zhang, Yu Cao, Dongdong Wang, Haifeng Dong, Xueji Zhang
As important modulators of gene expression, microRNAs (miRNAs) have been identified as promising biomarkers with powerful predictive value in diagnosis and prognosis for several diseases, especially for cancers. Here we report a facile, multiple and sensitive miRNA detection method that uses conventional gel electrophoresis and catalytic hairpin assembly (CHA) system without any complex nanomaterials or enzymatic amplification. Methods: In this study, three pairs of hairpin probes are rationally designed with thermodynamically and kinetically preferable feasibility for the CHA process...
2018: Theranostics
https://www.readbyqxmd.com/read/29770110/a-pcr-based-method-for-rna-probes-and-applications-in-neuroscience
#5
Ruifang Hua, Shanshan Yu, Mugen Liu, Haohong Li
In situ hybridization (ISH) is a powerful technique that is used to detect the localization of specific nucleic acid sequences for understanding the organization, regulation, and function of genes. However, in most cases, RNA probes are obtained by in vitro transcription from plasmids containing specific promoter elements and mRNA-specific cDNA. Probes originating from plasmid vectors are time-consuming and not suitable for the rapid gene mapping. Here, we introduce a simplified method to prepare digoxigenin (DIG)-labeled non-radioactive RNA probes based on polymerase chain reaction (PCR) amplification and applications in free-floating mouse brain sections...
2018: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/29768105/identification-of-a-recurrent-lmo7-braf-fusion-in-papillary-thyroid-carcinoma
#6
Huiling He, Wei Li, Pearlly Yan, Ralf Bundschuh, Jackson A Killian, Jadwiga Labanowska, Pamela Brock, Rulong Shen, Nyla A Heerema, Albert de la Chapelle
BACKGROUND: The BRAFV600E mutation is the most common driver in papillary thyroid carcinoma (PTC) tumors. In recent years, gene fusions have also been recognized as important drivers of cancer in PTC. Previous studies have suggested that thyroid tumors with fusion genes frequently display an aggressive course. These observations prompted further exploration of gene fusions in PTC tumors. The aim was to search for previously unrecognized gene fusions using thyroid tissue samples from PTC patients...
May 16, 2018: Thyroid: Official Journal of the American Thyroid Association
https://www.readbyqxmd.com/read/29768011/detection-of-sub-fm-dna-with-target-recycling-and-self-assembly-amplification-on-graphene-field-effect-biosensors
#7
Zhaoli Gao, Han Xia, Jonathan Zauberman, Maurizio Tomaiuolo, Jinglei Ping, Qicheng Zhang, Pedro Ducos, Huacheng Ye, Sheng Wang, Xinping Yang, Fahmida Lubna, Zhengtang Tom Luo, Li Ren, Alan T Charlie Johnson
All-electronic DNA biosensors based on graphene field effect transistors (GFETs) offer the prospect of simple and cost-effective diagnostics. For GFET sensors based on complementary probe DNA, the sensitivity is limited by the binding affinity of the target oligonucleotide, in the nM-range for 20-mer targets. We report a ~20,000× improvement in sensitivity through the use of engineered hairpin probe DNA that allows for target recycling and hybridization chain reaction. This enables detection of 21-mer target DNA at sub-fM concentration and provides superior specificity against single-base mismatched oligomers...
May 16, 2018: Nano Letters
https://www.readbyqxmd.com/read/29762748/molecular-type-specific-multiplex-pcr-produces-a-distinct-vnii-pcr-pattern-among-cryptococcus-neoformans-species-complex
#8
Massimo Cogliati, Mala Rakoto Andrianarivelo, Mohamed Ellabib, Emmanuel N Nnadi, Muriel Cornet
Prevalence of molecular type VNII among Cryptococcus neoformans species complex is probably underestimated since it can be distinguished from VNI only using molecular typing methods such as URA5-RFLP, AFLP, MLST, or whole genome sequencing. Previously, we described a multiplex polymerase chain reaction (PCR) method able to identify VNI, VNIV, and VNIII hybrids, but, at that time, VNII molecular type was not described yet. In this study, 16 VNII global isolates were analyzed by our multiplex PCR method, and results showed that it was able to produce a specific pattern for all the studied VNII isolates, which was different from those of VNI, VNIV, and VNIII...
May 14, 2018: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
https://www.readbyqxmd.com/read/29760303/mir-140-3p-is-involved-in-in-stent-restenosis-by-targeting-c-myb-and-bcl-2-in-peripheral-artery-disease
#9
Zheng-Rong Zhu, Qiong He, Wei-Bin Wu, Guang-Qi Chang, Chen Yao, Yang Zhao, Mian Wang, Shen-Ming Wang
AIM: In-Stent Restenosis (ISR) is the major reason for recurrent ischemia and amputation after endovascular treatment of Peripheral Artery Disease (PAD). Our previous study demonstrated that miR-140-3p is significantly down-regulated in PAD arteries. However, expression and function of miR-140-3p in ISR of human PAD are currently unclear. The aim of this study is to determine the miR-140-3p expression and its regulative role in ISR of PAD. METHODS: The RNA level was determined by quantitative real-time polymerase chain Reaction (qRT-PCR) and in situ hybridization...
May 11, 2018: Journal of Atherosclerosis and Thrombosis
https://www.readbyqxmd.com/read/29753166/double-determination-of-long-noncoding-rnas-from-lung-cancer-via-multi-amplified-electrochemical-genosensor-at-sub-femtomole-level
#10
Xiaoyan Li, Gang Peng, Feng Cui, Qianying Qiu, Xiaojun Chen, He Huang
An ultrasensitive electrochemical genosensor has been fabricated for the double determination of two different specific sequences deduced from the maternally expressed gene3 (MEG3) lncRNA (long noncoding RNA), which was demonstrated by coupling RNase A-aided target recycling with DNA supersandwich-induced signal enhancement, based on a composite interface of graphene-like tungsten disulfide/dendritic gold nanostructures (WS2 /DGN). Firstly, duple target sequences of T1 and T2 were captured by the primer probes of P1 /P2 functionalized Fe3 O4 @C magnetic nanoparticles, via the DNA/RNA hybridization between T1 /T2 and P1 /P2 ...
May 2, 2018: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29747453/chitinase-mrna-levels-determined-by-qpcr-in-crab-eating-monkey-macaca-fascicularis-tissues-species-specific-expression-of-acidic-mammalian-chitinase-and-chitotriosidase
#11
Maiko Uehara, Eri Tabata, Kazuhiro Ishii, Akira Sawa, Misa Ohno, Masayoshi Sakaguchi, Vaclav Matoska, Peter O Bauer, Fumitaka Oyama
Mice and humans express two active chitinases: acidic mammalian chitinase (AMCase) and chitotriosidase (CHIT1). Both chitinases are thought to play important roles in specific pathophysiological conditions. The crab-eating monkey ( Macaca fascicularis ) is one of the most frequently used nonhuman primate models in basic and applied biomedical research. Here, we performed gene expression analysis of two chitinases in normal crab-eating monkey tissues by way of quantitative real-time polymerase chain reaction (qPCR) using a single standard DNA molecule...
May 9, 2018: Genes
https://www.readbyqxmd.com/read/29739921/microrna-181a-promotes-angiogenesis-in-colorectal-cancer-by-targeting-srcin1-to-promote-the-src-vegf-signaling-pathway
#12
Wu Sun, Xiaojun Wang, Jialu Li, Chaoying You, Pan Lu, Huijin Feng, Yan Kong, Haiyang Zhang, Yanqing Liu, Ruihua Jiao, Xi Chen, Yi Ba
Colorectal cancer (CRC) is a very common metastatic tumor with active angiogenesis that requires active angiogenesis. Recently, increased microRNA-181a-5p (miR-181a) expression was found to be significantly associated with liver metastasis and poor outcome in CRC patients. In this study, the role of miR-181a in tumor angiogenesis was further investigated. Capillary tube formation assays were used to demonstrate the ability of miR-181a to promote tumor angiogenesis. Bioinformatics analyses identified SRC kinase signaling inhibitor 1 (SRCIN1) as a potential target of miR-181a...
April 19, 2018: Cell Death & Disease
https://www.readbyqxmd.com/read/29738818/microrna-132-targeting-pten-contributes-to-cilostazol-promoted-vascular-smooth-muscle-cell-differentiation
#13
Wei-Jan Chen, Ying-Hwa Chen, Yu-Juei Hsu, Kwang-Huei Lin, Yung-Hsin Yeh
BACKGROUND AND AIMS: Cilostazol, beyond its antiplatelet effect, is also capable of promoting vascular smooth muscle cell (VSMC) differentiation. The aim of this study was to explore the potential role of PTEN, known to associate with VSMC differentiation, and its related microRNA (miRNA) in cilostazol-dependent effects. METHODS AND RESULTS: Microarray analysis in balloon-injured rat carotid arteries comparing with and without balloon injury revealed that miR-132 was differentially expressed...
April 26, 2018: Atherosclerosis
https://www.readbyqxmd.com/read/29734245/human-epidermal-growth-factor-receptor-2-promotes-invasion-and-metastasis-in-gastric-cancer-by-activating-mitogen-activated-protein-kinase-signaling
#14
Feng Hou, Duan-Bo Shi, Yun-Qing Chen, Peng Gao
Increasing evidence supports an important role for the human epidermal growth factor receptor-2 (HER2) gene and mitogen-activated protein kinase (MAPK) signaling pathways in the progression of human cancers by enhancing cancer cell metastasis and proliferation. However, the relationship between HER2 and MAPK signaling pathways in gastric cancer (GC) remains unclear. In the present study, dual in situ hybridization was performed to detect HER2 gene amplification and reverse transcription-quantitative polymerase chain reaction was used to investigate the mRNA expression of members of the MAPK signaling pathway, including rapidly accelerated fibrosarcoma (RAF), extracellular regulated signal-activated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK), in 112 primary GC tissue samples...
May 4, 2018: Applied Immunohistochemistry & Molecular Morphology: AIMM
https://www.readbyqxmd.com/read/29733201/chemical-and-biological-sensing-using-hybridization-chain-reaction
#15
Erik E Augspurger, Muhit Rana, Mehmet Veysel Yigit
Since the advent of its theoretical discovery more than 30 years ago, DNA nanotechnology has been used in a plethora of diverse applications in both the fundamental and applied sciences. DNA-based technologies' recent prominence in the scientific community is largely due to the programmable features stored in its nucleobase composition and sequence, which allow it to assemble into highly advanced structures. DNA nanoassemblies are also highly controllable due to the precision of natural and artificial base-pairing, which can be manipulated by pH, temperature, metal ions and solvent types...
May 7, 2018: ACS Sensors
https://www.readbyqxmd.com/read/29725685/polymer-free-electrospinning-of-tannic-acid-and-cross-linking-in-water-for-hybrid-supramolecular-nanofibres
#16
Manon Allais, Domitille Mailley, Pascal Hébraud, Dris Ihiawakrim, Vincent Ball, Florent Meyer, Anne Hébraud, Guy Schlatter
Electrospinning is the process of choice allowing the preparation of nanofibrous materials from a solution usually based on a high molar mass polymer. The solution must bring enough chain entanglements to avoid any breaking or Rayleigh instability of the electrospun jet resulting thus in the deposition of a continuous and regular solid nanofibre. It has been however shown that some few non-polymeric molecules can be electrospun without using a carrier polymer. We demonstrate here the case of tannic acid. Indeed, it was possible to electrospin this molecule solubilised in a mixture of water and ethanol as well as in pure water...
May 4, 2018: Nanoscale
https://www.readbyqxmd.com/read/29722265/near-infrared-fluorescent-ag2s-nanodot-based-signal-amplification-for-efficient-detection-of-circulating-tumor-cells
#17
Caiping Ding, Cuiling Zhang, Xueyang Yin, Xuanyu Cao, Meifang Cai, Yuezhong Xian
The level of circulating tumor cells (CTCs) plays a critical role in tumor metastasis and personalized therapy, but it is chal-lenging for highly efficient capture and detection of CTCs because of the extremely low concentration in peripheral blood. Herein, we report near-infrared fluorescent Ag2S nanodot-based signal amplification combing with immune-magnetic spheres (IMNs) for highly efficient magnetic capture and ultrasensitive fluorescence labeling of CTCs. The near-infrared fluorescent Ag2S nanoprobe has been successfully constructed through hybridization chain reactions using aptamer modi-fied Ag2S nanodots, which can extremely improve the imaging sensitivity and reduce background signal of blood samples...
May 3, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29720215/dependence-receptor-unc5a-restricts-luminal-to-basal-breast-cancer-plasticity-and-metastasis
#18
Maria B Padua, Poornima Bhat-Nakshatri, Manjushree Anjanappa, Mayuri S Prasad, Yangyang Hao, Xi Rao, Sheng Liu, Jun Wan, Yunlong Liu, Kyle McElyea, Max Jacobsen, George Sandusky, Sandra Althouse, Susan Perkins, Harikrishna Nakshatri
BACKGROUND: The majority of estrogen receptor-positive (ERα+ ) breast cancers respond to endocrine therapies. However, resistance to endocrine therapies is common in 30% of cases, which may be due to altered ERα signaling and/or enhanced plasticity of cancer cells leading to breast cancer subtype conversion. The mechanisms leading to enhanced plasticity of ERα-positive cancer cells are unknown. METHODS: We used short hairpin (sh)RNA and/or the CRISPR/Cas9 system to knockdown the expression of the dependence receptor UNC5A in ERα+ MCF7 and T-47D cell lines...
May 2, 2018: Breast Cancer Research: BCR
https://www.readbyqxmd.com/read/29718508/expression-characterization-and-localization-of-the-foraging-gene-in-the-chinese-bee-apis-cerana-cerana-hymenoptera-apidae
#19
WeiHua Ma, YuSuo Jiang, Jiao Meng, HuiTing Zhao, HuaiLei Song, JinShan Shen
In social insects, the foraging gene (for) regulates insect age- and task-based foraging behaviors. We studied the expression and localization of the for gene (Acfor) in Apis cerana cerana workers to explore whether the differential regulation of this gene is associated with the behaviors of nurses and foragers. The expression profiles of Acfor in different tissues and at different ages were examined using real-time quantitative reverse transcription polymerase chain reaction. Cellular localization in the brain was detected using in situ hybridization...
March 1, 2018: Journal of Insect Science
https://www.readbyqxmd.com/read/29717053/touchdown-polymerase-chain-reaction-pcr
#20
Michael R Green, Joseph Sambrook
"Touchdown polymerase chain reaction (PCR)" is a method to decrease off-target priming and hence to increase the specificity of PCRs. In touchdown PCR the temperature selected for the annealing step is initially set 5°C-10°C higher than the calculated T m of the primers. Annealing under conditions of high stringency favors the formation of perfect primer-template hybrids. In subsequent cycles, the annealing temperature is gradually decreased by a small amount so that by the end of the PCR, the annealing temperature is 2°C-5°C below the calculated T m of the primers...
May 1, 2018: Cold Spring Harbor Protocols
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