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High resolution melting

Reut Shapira, Yael S Balazs, Shifi Kababya, Rachel Edrei, Yoav Eichen
Inverse melting α-cyclodextrin·4-methylpyridine·H2O ternary mixtures were studied via solution, high-resolution magic-angle-spinning and 2D-NOESY NMR spectroscopy with the aim of unveiling the supramolecular driving force for the unusual inverse-melting phase transition. The role of solvent-solvent interactions as well as solvent-solute interactions in the emergence of the inverse-melting phenomenon is revealed. A surprising re-entrant solvent-solvent interaction is found in the system and in the 4-methylpyridine·water solvent system, supporting the non-coincidental relationship between inverse melting and re-entrant phase transitions, at least in the present system...
October 9, 2018: Physical Chemistry Chemical Physics: PCCP
Rafael Amorim Belo Nunes, Larissa Barbosa Lima, Nelson Ithiro Tanaka, Alexandre da Costa Pereira, José Eduardo Krieger, Alfredo José Mansur
Background: Physiological pathways such as bradykinin, renin-angiotensin, neurohormones and nitric oxide have been shown to play an important role in the regulation of cardiovascular function. Genetic variants of these pathways may impact blood pressure and left ventricular (LV) mass in different populations. To evaluate associations of genetic polymorphisms of bradykinin B2 receptor (BDKRB2), alpha-adrenergic receptors (ADRA) and endothelial nitric oxide synthase (eNOS) on the modulation of the blood pressure and the left ventricular mass...
December 2018: IJC Heart & Vasculature
H Zhang, Y P Liu, A Q Ge, X Wang, H R Sun, H R Bi, D Pang, Y S Zhao
Objective: To understand the relationship between AOX1, IRF4 gene methylation status in peripheral blood leukocyte DNA, as well as its interaction with environmental factors, and the risk of breast cancer. Methods: A case-control study was conducted among 401 breast cancer patients and 555 cancer-free controls selected from 2010 to 2014. Methylation sensitive-high resolution melting curve analysis was used to detect the methylation status of AOX1 and IRF4. The multiplication interaction effect between genes' methylation and environmental factors on the risk of breast cancer was analyzed by using unconditional logistic regression, and Excel software was used to analyze the additive interaction effect...
September 10, 2018: Zhonghua Liu Xing Bing Xue za Zhi, Zhonghua Liuxingbingxue Zazhi
Juraj Sokol, Maria Skerenova, Kamil Biringer, Tomas Simurda, Peter Kubisz, Jan Stasko
The aim of our study was to evaluate GP6 gene in patients with sticky platelet syndrome (SPS) and fetal loss. Platelet aggregability was tested with platelet-rich plasma using PACKS-4 aggregometer (Helena Laboratories). High-resolution melting analysis on LightCycler 480 II (Roche Diagnostics) was used for single-nucleotide polymorphism (SNP) genotyping. We examined 64 patients with SPS and 54 control participants. We found significantly higher occurrence of 5 SNPs in patients with SPS versus controls (rs1671152, rs1654433, rs1613662, rs1654416, and rs2304167)...
October 2, 2018: Clinical and Applied Thrombosis/hemostasis
Xin Zhao, Weilan Wang, Alysson Blaine, Sarah T Kane, Ruurd T Zijlstra, Michael G Gänzle
AIMS: This study aimed to determine whether host-adapted lactobacilli exhibit superior survival during intestinal transit relative to nomadic and free-living organisms, and to characterize the impact of probiotic lactobacilli on autochthonous lactobacilli. METHODS AND RESULTS: Mixed cultures of Lactobacillus casei K9-1 and Lactobacillus fermentum K9-2, or reutericyclin producing L. reuteri and its isogenic mutant were fed to piglets as freeze-dried culture, or as part of fermented feed...
October 1, 2018: Journal of Applied Microbiology
Subhabrata Majumder, Michael T Jones, Michael Kimmel, Arun Alphonse Ignatius
PURPOSE: Fc domains are an integral component of monoclonal antibodies (mAbs) and Fc-based fusion proteins. Engineering mutations in the Fc domain is a common approach to achieve desired effector function and clinical efficacy of therapeutic mAbs. It remains debatable, however, whether molecular engineering either by changing glycosylation patterns or by amino acid mutation in Fc domain could impact the higher order structure of Fc domain potentially leading to increased aggregation propensities in mAbs...
September 25, 2018: Pharmaceutical Research
Timothy S Carpenter, Cesar A López, Christopher Neale, Cameron Montour, Helgi Ingólfur Ingólfsson, Francesco Di Natale, Felice C Lightstone, Sandrasegaram Gnanakaran
The presence of lipid rafts in the membranes of living cells remains hotly disputed despite their incontrovertible existence in liposomes at 298 K. In attempts to resolve this debate, molecular dynamics (MD) simulations have been extensively used to study lipid phase separation at high resolution. However, computation has been of limited utility in this respect because the experimental distributions of phases in lamellar lipid mixtures are poorly reproduced by simulations. In particular, all-atom (AA) approaches suffer from restrictions on accessible timescales and system sizes whereas the more efficient coarse-grained (CG) force fields remain insufficiently accurate to achieve correspondence with experiment...
September 25, 2018: Journal of Chemical Theory and Computation
Chloe Victoria Robinson, Tamsyn M Uren Webster, Sofia Consuegra
The data presented here corresponds to the research paper "Simultaneous detection of invasive signal crayfish, endangered white-clawed crayfish and the crayfish plague using environmental DNA". A crayfish-specific assay was designed and optimised using three real-time PCR supermixes (SYBR™ Green, SsoFast™ EvaGreen® and HOT FIREPol® EvaGreen®). Diagnostic high resolution melt (HRM) data from direct application of assay on both ex-situ eDNA water samples and field samples from four catchments (two in Wales, two in England) is presented in this article, displaying positive HRM profiles for invasive signal crayfish ( Pacifastacus leniusculus), native white-clawed crayfish ( Austropotamobius pallipes ) and crayfish plague causal agent ( Aphanomyces astaci )...
August 2018: Data in Brief
José Javier Marqueta-Gracia, Maite Álvarez-Álvarez, Miriam Baeta, Leire Palencia-Madrid, Endika Prieto-Fernández, Juan Ramón Ordoñana, Marian M de Pancorbo
Discrimination between monozygotic (MZ) twins is a forensic limitation when using conventional DNA profiling techniques for human identification. Recent works based on epigenetics seem to open a new way to solve this issue due to methylation status of MZ twins change during their lifetime. Methylation analysis through BeadChip platforms allows the study up to 850 K CpG sites revealing that numerous differential methylation regions exist between MZ twins. However, this methodology is difficult to implement in forensic laboratories...
September 20, 2018: Forensic Science International. Genetics
Ming-Jr Jian, Yun-Hsiang Cheng, Cherng-Lih Perng, Hung-Sheng Shang
Objectives: Several Elizabethkingia species often exhibit extensive antibiotic resistance, causing infections associated with severe morbidity and high mortality rates worldwide. In this study, we determined fluoroquinolone susceptibility profiles of clinical Elizabethkingia spp. isolates and investigated the resistance mechanisms. Methods: In 2017-2018, 131 Elizabethkingia spp. isolates were recovered from specimens collected at tertiary care centers in northern Taiwan...
2018: PeerJ
Xiaoming Chen, Zhuoqing Hu, Meilian Liu, Huaqian Li, Chanbo Liang, Wei Li, Liwen Bao, Manyang Chen, Ge Wu
BACKGROUND: Single-nucleotide polymorphism (SNP) haplotype and SNP-SNP interactions of CTLA-4 and CD40 genes, with susceptibility to Graves' disease (GD), were explored in a Chinese Han population. METHODS: SNP were genotyped by high resolution melting (HRM). Use the method of Pearson χ2 test and Logistic regression for the association between single SNP and Graves' disease. Using the method of χ2 test and Multifactor Dimensionality Reduction (MDR) to analysis the haplotype frequency distribution, the interaction of SNPs respectively...
September 17, 2018: BMC Medical Genetics
Marta Ferro, Hada C Macher, Gema Fornés, Jesús Martín-Sánchez, Pilar Jimenez-Arriscado, Patrocinio Molinero, José A Pérez-Simón, Juan M Guerrero, Amalia Rubio
BACKGROUND: The development of new noninvasive approaches for the diagnosis of human platelet antigen (HPA)-1 fetomaternal incompatibility has become of great interest. These approaches allow determination of whether the fetus is incompatible or not with the mother and a decision on antenatal therapy to avoid fetal or neonatal alloimmune thrombocytopenia (FNAIT). The objective of this work was to perform rapid, noninvasive prenatal test for HPA-1ab fetal antigen detection after the detection of an HPA-1-homozygous mother by using plasma cell-free DNA (cfDNA)...
October 2018: Transfusion
Kenneth E Sherman, Susan D Rouster, Minhee Kang, Triin Umbleja, Richard Sterling, Adeel A Butt
BACKGROUND AND AIMS: The patatin-like phospholipase domain-containing 3 (PNPLA3) gene has been associated with the development of alcoholic and nonalcoholic steatohepatitis. Using a newly developed and validated assay for PNPLA3, we explored the prevalence of gene polymorphisms in a cohort of HCV/HIV-coinfected individuals to determine whether there was an association with insulin resistance or hepatic fibrosis. METHODS: A high-resolution melting point (HRM) assay was developed and validated...
September 14, 2018: Digestive Diseases and Sciences
Sumana Roy, Sudeshna Das Chakraborty, Sampa Biswas
Human cathepsin K (CTSK) is a collagenolytic lysosomal cysteine protease that plays an important role in bone turnover. Mutation in CTSK gene is associated with loss of collagenolytic activity of CTSK leading to an autosomal recessive bone disorder called pycnodysostosis. Although a number of pycnodysostotic missense mutations have been reported, underlying mechanism of the disease is not clear. In this study, we investigated in vitro six recombinant pycnodysostosis-related mutants of human CTSK (G79E, I249T, G243E, G303E, G319C and Q187P)...
September 10, 2018: FEBS Journal
Sanjay Singh Negi, Priyanka Singh, Anudita Bhargava, Sachin Chandrakar, Ujjwala Gaikwad, Padma Das, Ajoy Behra
Background: Effective management of multidrug-resistant tuberculosis (MDR-TB) requires cost-effective and rapid screening of rifampicin (RIF) and isoniazid (INH) resistance. Accordingly, a highly promising high-resolution melting (HRM) analysis was evaluated in the detection of mutation in rpoB, katG gene and inhA promoter region in Mycobacterium tuberculosis isolates. Methods: A total of 143 M. tuberculosis isolates comprising phenotypically confirmed 94 MDR and 49 sensitive isolates were analyzed by HRM following real-time-polymerase chain reaction in comparison to gold standard of targeted DNA sequencing of rpoB, katG gene and inhA promoter region...
July 2018: International Journal of Mycobacteriology
Junlin Chen, Fei Huang, Xi Yin, Delin Gu
This study was undertaken to compare the ability of four clinically used methods to detect the heteroresistance of the tubercle bacillus to Rifampicin (Rif). A total of 94 sputum smear-positive samples of re-treated tuberculosis (TB) patients were employed. Rif-resistant tubercle bacillus tests were carried out by four methods: Lowenstein-Jensen cultures (L-J), reverse transcription-polymerase chain reaction (RT-PCR), microscopic observation drug susceptibility (MODS), and high-resolution melting (HRM). The rates of detection of Rif-resistant bacteria by of L-J, RT-PCR, MODS, and HRM were 35...
December 1, 2018: Journal of Nanoscience and Nanotechnology
Manuela Tamburro, Michela Lucia Sammarco, Giancarlo Ripabelli
AIMS: A High Resolution Melting (HRM) assay was developed for characterizing lineage II Listeria monocytogenes based on the amplification and the melting profiles analysis of 81 fragments targeting the region from the prs to ldh loci, including the Listeria Pathogenicity Island-1 (LIPI-1) genes, and the inlAB operon. METHODS AND RESULTS: Real-time PCR and HRM protocols were standardized using 10 replicate assays from L. monocytogenes EGD-e reference strain (serovar 1/2a)...
September 6, 2018: Journal of Applied Microbiology
Iben Daugaard, Dianna Hussmann, Louise Kristensen, Thomas Kristensen, Tina E Kjeldsen, Charlotte G Nyvold, Thomas S Larsen, Michael B Møller, Lise Lotte Hansen, Tomasz K Wojdacz
AIM: We investigated whether DNA methylation regulates expression of LPL and PI3K complex genes in chronic lymphocytic leukemia (CLL) and evaluated the prognostic significance of LPL promoter methylation in CLL patients. Patients & methods: Methylation of LPL promoter was assessed in 112 patients using methylation-sensitive high-resolution melting (MS-HRM). RESULTS: Patients with a fully or heterogeneously methylated LPL promoter had significantly longer median time to treatment (p < 0...
September 2018: Epigenomics
Niranjan D Parab, Cang Zhao, Ross Cunningham, Luis I Escano, Kamel Fezzaa, Wes Everhart, Anthony D Rollett, Lianyi Chen, Tao Sun
The high-speed synchrotron X-ray imaging technique was synchronized with a custom-built laser-melting setup to capture the dynamics of laser powder-bed fusion processes in situ. Various significant phenomena, including vapor-depression and melt-pool dynamics and powder-spatter ejection, were captured with high spatial and temporal resolution. Imaging frame rates of up to 10 MHz were used to capture the rapid changes in these highly dynamic phenomena. At the same time, relatively slow frame rates were employed to capture large-scale changes during the process...
September 1, 2018: Journal of Synchrotron Radiation
Jianying Zhang, Tao Zhou, Yichuan Tang, Yanjie Cui, Dan Song
High-purity silicon power was doped with standard solutions containing 15 elements, and a high-purity indium tablet was prepared by the melting of indium pellets. An In-Si tablet, which is mechanically stable and thus suitable as a calibration sample, was prepared by our pressing the doped silicon power on the high-purity indium tablet. The matrix effect was studied by our investigating the variations of measured mass fractions (standard relative sensitivity factor, StdRSF, calibration) of doped impurities in the In-Si tablet, which provides a series of matrixes with different mass ratios of In to Si...
September 3, 2018: Analytical and Bioanalytical Chemistry
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