Loop mediated isothermal amplification (LAMP) is a nucleic acid amplification technique performed under isothermal conditions. The output of this amplification technique includes multiple different sizes of deoxyribonucleic acid (DNA) structures which are identified by a banding pattern on gel electrophoresis plots. Although this is a specific amplification technique, the complexity of the primer design and amplification still lead to the issue of obtaining false‐positive results, especially when a positive reading is determined solely by whether there is any banding pattern in the gel electrophoresis plot. Here, we first performed extensive LAMP experiments and evaluated the DNA structures using microchip electrophoresis. We then developed a mathematical model derived from the various components that make up an entire LAMP structure to predict the full LAMP structure size in base pairs. This model can be implemented by users to make predictions for specific, DNA size dependent, banding patterns on their gel electrophoresis plots. Each prediction is specific to the target sequence and primers used and therefore reduces incorrect diagnosis errors through identifying true‐positive and false‐positive results. This model was accurately tested with multiple primer sets in house and was also translatable to different DNA and RNA types in previously published literature. The mathematical model can ultimately be used to reduce false‐positive LAMP diagnosis errors for applications ranging from tuberculosis diagnostics to E. coli to numerous other infectious diseases. 相似文献
The purpose of this study was to determine the value of Gradient Echo imaging for the evaluation of cartilage (3D fatsat) and blood products (2D Hemoflash), and the use of contrast enhanced SE imaging for the evaluation of synovial changes, in comparison to the clinical evaluation of children with hemophilia A. We investigated 21 joints in 16 patients with evidence of hemophilia A (mean age 11.3+/-2.1 years). In all patients, clinical examination, plain film radiographs, and MR evaluation were performed magnetic resonance imaging (MRI) was performed by using sagittal T1 SE and T2 SE images, as well as 3D fatsat GE and 2D GE images. Axial and sagittal T1 weighted SE images were obtained before and after contrast application. Findings from the clinical examination and MR imaging, regarding the evaluation of blood, synovia, and cartilage were compared. Clinical examination revealed evidence of a bleeding episode in 12 joints (57.1%), whereas MRI revealed evidence of blood or blood products in 15 joints (71.4%). Clinical investigations, including bleeding scores, pain scores, and physical examination scores did not correlate with MR findings. Due to the MR findings in 6 of 16 patients, therapeutic management was changed from on demand to prophylactic therapy. MR imaging with gradient echo and contrast-enhanced sequences is more sensitive than clinical examination for the detection of blood products in children with hemophilia. Its ability to demonstrate potentially early stages of cartilage or synovial alterations might assist in therapy planning. Clinical scores might underestimate effects of hemophilia. 相似文献
An enantiomerically pure diamine based on the 4,15-difunctionalized [2.2]paracyclophane scaffold and 2-formylpyridine self-assemble into an optically pure cyclic metallosupramolecular Fe4L6 helicate upon mixing with iron(II) ions in a diastereoselective subcomponent self-assembly process. The cyclic assembly results from steric strain that prevents the formation of a smaller linear dinuclear triple-stranded helicate, and hence, leads to the larger strain-free assembly that fulfils the maximum occupancy rule. Interestingly, use of the racemic diamine also leads to a racemic mixture of the homochiral cyclic helicates as the major product in a highly diastereoselective narcissistic chiral self-sorting manner given the fact that the assembly contains ten stereogenic elements, which can in principle give rise to 149 different diastereomers. The metallosupramolecular aggregates could be characterized by NMR, UV/Vis and CD spectroscopy, mass spectrometry, and X-ray crystallography. 相似文献
Organodihydridoelement anions of germanium and tin were reacted with metallocene dichlorides of Group 4 metals Ti, Zr and Hf. The germate anion [Ar*GeH2]− reacts with hafnocene dichloride under formation of the substitution product [Cp2Hf(GeH2Ar*)2]. Reaction of the organodihydridostannate with metallocene dichlorides affords the reduction products [Cp2M(SnHAr*)2] (M=Ti, Zr, Hf). Abstraction of a hydride substituent from the titanium bis(hydridoorganostannylene) complex results in formation of cation [Cp2M(SnAr*)(SnHAr*)]+ exhibiting a short Ti–Sn interaction. (Ar*=2,6-Trip2C6H3, Trip=2,4,6-triisopropylphenyl). 相似文献
Histamine H4 receptor (H4R) orthologues are G-protein-coupled receptors (GPCRs) that exhibit species-dependent basal activity. In contrast to the basally inactive mouse H4R (mH4R), human H4R (hH4R) shows a high degree of basal activity. We have performed long-timescale molecular dynamics simulations and rigidity analyses on wild-type hH4R, the experimentally characterized hH4R variants S179M, F169V, F169V+S179M, F168A, and on mH4R to investigate the molecular nature of the differential basal activity. H4R variant-dependent differences between essential motifs of GPCR activation and structural stabilities correlate with experimentally determined basal activities and provide a molecular explanation for the differences in basal activation. Strikingly, during the MD simulations, F16945.55 dips into the orthosteric binding pocket only in the case of hH4R, thus adopting the role of an agonist and contributing to the stabilization of the active state. The results shed new light on the molecular mechanism of basal H4R activation that are of importance for other GPCRs. 相似文献
For the last decades, the hydrogen-abstraction−acetylene-addition (HACA) mechanism has been widely invoked to rationalize the high-temperature synthesis of PAHs as detected in carbonaceous meteorites (CM) and proposed to exist in the interstellar medium (ISM). By unravelling the chemistry of the 9-phenanthrenyl radical ([C14H9].) with vinylacetylene (C4H4), we present the first compelling evidence of a barrier-less pathway leading to a prototype tetracyclic PAH – triphenylene (C18H12) – via an unconventional hydrogen abstraction–vinylacetylene addition (HAVA) mechanism operational at temperatures as low as 10 K. The barrier-less, exoergic nature of the reaction reveals HAVA as a versatile reaction mechanism that may drive molecular mass growth processes to PAHs and even two-dimensional, graphene-type nanostructures in cold environments in deep space thus leading to a better understanding of the carbon chemistry in our universe through the untangling of elementary reactions on the most fundamental level. 相似文献
Immunological methods are widely applied in medical diagnostics for the detection and quantification of a plethora of analytes. Associated analytical challenges usually require these assays to be performed in a central laboratory. During the last several years, however, the clinical demand for rapid immunodiagnostics to be performed in the immediate proximity of the patient has been constantly increasing. Biosensors constitute one of the key technologies enabling the necessary, yet challenging transition of immunodiagnostic tests from the central laboratory to the point of care. This review is intended to provide insights into the current state of this transition process with a focus on the role of biosensor-based systems. To begin with, an overview on standard immunodiagnostic tests presently employed in the central laboratory and at the point of care is given. The review then moves on to demonstrate how biosensor technologies are reshaping this landscape. Single analyte as well as multiplexed immunosensors applicable to point of care scenarios are presented. A section on the areas of clinical application then creates the bridge to day-to-day diagnostic practice. Finally, the depicted developments are critically weighed and future perspectives discussed in order to give the reader a firm idea on the forthcoming trends to be expected in this diagnostic field.
We report results of molecular dynamics simulations characterizing the solvation and interaction of small apolar particles such as methane and xenon in imidazolium-based ionic liquids (ILs). The simulations are able to reproduce semiquantitatively the anomalous temperature dependence of the solubility of apolar particles in the infinite dilution regime. We observe that the "solvophobic solvation" of small apolar particles in ILs is governed by compensating entropic and enthalpic contributions, very much like the hydrophobic hydration of small apolar particles in liquid water. In addition, our simulations clearly indicate that the solvent mediated interaction of apolar particles dissolved in ILs is similarly driven by compensating enthalpic and entropic contributions, making the "solvophobic interaction" thermodynamically analogous to the hydrophobic interaction. 相似文献