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[en] Highlights: • A novel colorimetric strategy for sensitive DNA detection based on AuNPs assemblies induced by PCR product. • The proposed strategy avoids the denaturation of PCR product to allow efficient hybridization with AuNPs-DNA probes. • The sensitivity of this method is significantly improved, due to PCR amplification and the assemblies of AuNPs. • The universality of the method is likely to be good, any target DNA can be analyzed with the same AuNPs-DNA probes. - Abstract: We developed a novel strategy for rapid colorimetric detection of specific DNA sequence based on gold nanoparticles assemblies induced by polymerase chain reaction (PCR) product. In the presence of target DNA, the two DNA-functionalized AuNP probes selectively hybridized with the prohibited nucleic acid segments of two primers owing to the zipping off of the hairpin structures during PCR process, resulted in the aggregation of AuNPs with a concomitant color change from red to blue-purple. It is a convenient and universal method for sensitive DNA detection with no need for any further post-treatment of the PCR products. Most importantly, our method showed a low limit of detection (LOD) of 4.3 fM with a wide range of target DNA from 16 fM to 1.6 nM. Owing to the versatility and low cost, the proposed strategy could be extremely useful for a wide range of applications, providing a promising tool for rapid disease diagnostics and gene sequencing.
[en] The results of studies into microporous scaffolds based on polycaprolactone, in particular, involving nanoparticles and microparticles of modified (silicon-containing) hydroxyapatite (hybrid scaffolds) are presented. When hydroxyapatite particles are used during the electrospinning of polymer scaffolds, their porosity is found to increase substantially and a structure with nanofibers and microfibers can be created. X-ray phase analysis demonstrates that the characteristic lines of polycaprolactone and hydroxyapatite exist in the 3D hybrid scaffold structure. According to the data of infrared (IR) spectroscopy of the hydroxyapatitepowder precursor, (SiO4)4– ions are embedded in its lattice. The results of studies into the surface wettability indicate that the contact angles of wetting with water are smaller for hybrid scaffolds than for pure polycaprolactone scaffolds. Adhesive and proliferative activity tests of human mesenchymal stem cells (MSCs) performed upon hybrid-scaffold cultivation on the surface, as well as histologic investigations, indicate the high biocompatibility of the samples. On the basis of a polymerase chain reaction, it is revealed that the differentiation of MSCs occurs in the osteogenic direction. On account of a porous structure, hybrid scaffolds can be employed to recover bone-tissue defects.
[en] Mass defect and binding energy, droplet model, energy balance, chain reaction, water as moderator and coolant, power control, afterheat. (orig.)
[de]Massendefekt und Bindungsenergie, Troepfchenmodell, Energiebilanz, Kettenreaktion, Wasser als Moderator und Kuehlmittel, Leistungsregelung, Nachwaerme. (orig.)
[en] Full text:The result of experimental and theoretical studies of radiation-thermal decomposition of heptanes admixtures in the water medium are adduced. The main parameters of radiolysis changed within the range: temperature 40-4000C, absorbed, dose-0-10.8 kGy on dose rate 3.6 kGy/h. As a product of decomposition are observed H2 CO, CH4, C2H4, C2H6, C3H8, C3H6, C4H8, traces of hydrocarbons C5, H6. The changes of n.heptane concentration in the reactor also were established. The chain regime of n. heptane decomposition in irradiated mixture is observed. The critical meaning of temperature and rate of components concentration, on which the break of chain process of heptanes decomposition is defined was observed.
[en] Background: Bleeding episodes commonly occur in patients on warfarin treatment even in those within therapeutic range of international normalized ratio (INR). The objective of this study was to investigate the effects of the 8 examined polymorphisms on the risk of bleeding complications in a sample of Iranian patients. Methods: A total of 552 warfarin treated patients who maintained on a target INR level of 2.0–3.5 for at least three consecutive intervals were enrolled from those attended our anticoagulation clinics. Ninety-two bleeding events were observed in 87 patients. The presences of the examined polymorphisms were analyzed using polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP). Results: Patients with the T allele in NQO1*2 (CT or TT genotypes) had a higher risk of bleeding than patients with the CC genotype (adjusted OR: 2.25, 95% CI: 1.37 to 3.70, P = 0.001). Those who were carriers of CYP2C9 one-variant haplotypes (*1/*2 or *1/*3) were also found to be associated with the higher risk of bleeding events. Compared to reference group (*1/*1), the odds of bleeding increased for carriers of one variant allele (*1/*2 or *1/*3) (adjusted OR: 1.75, 95% CI: 1.03 to 2.97, P = 0.039). Variant VKORC1, Factor VII, and EPHX1 genotypes were not significantly associated with the risk of bleeding events. Conclusion: The SNP C609T within NQO1 and haplotypes of CYP2C9 (1*2 or 1*3) are independently associated to bleeding complications of warfarin at normal INR. Further studies are required to confirm such associations in diverse racial and ethnic populations. - Highlights: • NQO1 C609T variant is associated with warfarin induced bleeding at therapeutic INR. • Haplotypes of CYP2C9 (1*2 or 1*3) are also associated with bleeding events. • VKORC1, Factor VII, and EPHX1 genotypes were not associated with bleeding risk.
[en] Tuberculosis remains a major international health problem despite advances in radiological diagnosis and antituberculous therapy. Disseminated tuberculous infection affecting a single bone and manifested as multifocal lytic cortical lesions is rare and unusual. We report on a 50-year-old man with multifocal involvement of the femur by tuberculosis and demonstrate positive reaction using tuberculosis-polymerase chain reaction on formalin-fixed, paraffin-embedded histological specimens. (orig.)
[en] Highlights: • Conventional multiplex PCR was combined with GE-ICP-MS for CNV and gene expression analyses in cancer cells. • Amplicons were separated, identified and label-free quantitate through 31P monitoring. • CNVs results obtained with this method were in accordance to literature data and to uniplex qPCR results. • This method provided the same information on relative gene expression for three genes, as single reaction RT-qPCR. - Abstract: During the last few years multiplex real-time or quantitative polymerase chain reaction PCR (qPCR) has become the method of choice for multiplex gene expression changes and gene copy number variations (CNVs) analysis. However, such determinations require the use of different fluorescent labels for the different amplified sequences, which increases significantly the costs of the analysis and limits the applicability of the technique for simultaneous amplification of many targets of interest in a single reaction. In this regard, the use of the coupling between gel electrophoresis (GE) separation with inductively coupled plasma mass spectrometry (ICP-MS) detection allows the label-free determination of multiplex PCR-amplified sequences (amplicons) by monitoring the P present in the DNA backbone. The quantitative dimension is obtained since under optimal and controlled multiplex PCR conditions the peak areas of the separated amplicons are directly proportional to the amount of DNA template in the original sample. Moreover, the calibration of the GE-ICP-MS system with a DNA ladder permits direct estimation of the size (bp) of the PCR products. The suitability of the proposed multiplex strategy has been evaluated addressing two different situations: determination of CNVs and gene expression changes in human ovarian cancer cells. In the first case, the results obtained for the simultaneous quantitation of CNVs of four genes (HER2, CCNE1, GSTM1, ACTB) on DNA obtained from OVCAR-3 cells were in accordance with the literature data, and also with the results obtained by conventional simplex qPCR. In the second case, multiplex gene expression changes of BAX, ERCC1 and CTR1 genes, using ACTB as constitutive gene, on A2780cis respect to A2780 cells, resistant and sensitive to cisplatin, respectively, provided the same information as single reaction reverse transcription (RT)-qPCR.
[en] Analytical method for solving radioactive transformations is presented in this paper. High accuracy series expansion of the depletion function and nonsingular Bateman coefficients are used to overcome numerical difficulties when applying well-known Bateman solution of a simple radioactive decay. Generality and simplicity of the method are found to be useful in evaluating nuclide chains with one hundred or more nuclides in the chain. Method enables evaluation of complete chain, without elimination of short-lives nuclides. It is efficient and accurate
[en] Human spinal cord injury (SCI) usually causes irreversible disability beneath the injured site due to poor neural regeneration. On the contrary, zebrafish show significant regenerative ability after SCI, thus is usually worked as an animal model for studying neuroregeneration. Most of the previous SCI studies focused on the local site of SCI, the supraspinal-derived signals were rarely mentioned. Here we showed that intradiencephalon injection of histamine (HA) inhibited the locomotor recovery in adult zebrafish post-SCI. Immunofluorescence results showed that intradiencephalon HA administration increased the activated microglia 3 days post injury (dpi), promoted the proliferation of radial glial cells at 7 dpi and affected the morphology of radial glial cells at 11 dpi. Furthermore, quantitative real-time polymerase chain reaction (qRT-PCR) results showed that intradiencephalon HA administration also reduced the expression of neurotrophic factors including brain-derived neurotrophic factor (BDNF) and insulin-like growth factor1 (IGF-1) at the lesion site, however, had no effect on the expression of pro-inflammatory factors such as TNF-alpha and IL-1 beta. Hence, our data suggested that exogenous intradiencephalon HA retarded locomotor recovery in spinal cord injured zebrafish via modulating the repair microenvironment. - Highlights: • A novel model to study the effect of supraspinal signals on repair of SCI zebrafish. • Exogenous HA administration suppresses the locomotor function recovery after SCI. • Exogenous HA administration modulates the repair microenvironment after SCI.
[en] The sensitivity of polymerase chain reaction (PCR) can mean that even very low levels of contamination with the target DNA will result in a positive signal. At present this aspect is a major limitation in the use of PCR as a routine diagnostic method. By exposing PCR reagents to UV light, contaminating DNA can be inactivated, thus providing an opportunity to eradicate false positive reactions. UV irradiation was applied to PCR systems used for detection of human cytomegalovirus CMV and human immunodeficiency virus (HIV) and shown to be effective in eradicating both laboratory encountered contamination and plasmid DNA (below 100 pg) added to PCR systems prior to UV exposure. Sensitivity of a PCR system to amplify the long terminal repeat (LTR) sequence of HIV-1 was not affected by the irradiation procedure; however, ultimate sensitivity of a PCR system for the amplification of an early gene pro-motor sequence of the CMV genome was reduced 1000-fold. UV irradiation did not affect the size of the PCR product as determined by strand separating polyacrylamide gel electrophoresis of a 32P-labelled amplimer. Thus, a simple pre-exposure to UV light would seem a worth-wile step to incorporate into PCR protocols provided that the effects on sensitivity have been determined empirically for each PCR system. (author). 11 refs.; 3 figs