Nucleic acid structure and sequence probing using fluorescent base analogue tC^o

The fluorescent cytosine analog tC(o) is on average the brightest probe of its kind and, moreover, it introduces minimal perturbations to the normal secondary structure of DNA. Here several ways of how tC(o), with an advantage, can be used as a local fluorescent probe in nucleic acid systems are presented. Most importantly, we show that tCo is an excellent probe for the detection of individual melting processes of complex nucleic acid structures containing a large number of separate secondary structure motifs. Since conventional UV-melting investigations merely monitor the global melting process of the whole nucleic acid structure, e.g. multi-hairpin systems in RNA/DNA, and thus is incapable of estimating individual melting transitions of such systems, tC(o) represents a new method of characterization. Furthermore, we find that tCo may be used to detect bulges and loops in nucleic acids as well as to distinguish a matched base-pair from several of the mismatched

Ökad tillämpning av ledningssystem i projekt

Att skapa en förståelse för hur företags ledningssystem kan tillämpas i verksamheten. Studien ska även finna en lösning för hur ledningssystem kan presenteras i ett webbaserat projektstyrningsverktyg för att skapa mervärde för användarna

FRET enhancement close to gold nanoparticles positioned in DNA origami constructs

Here we investigate the energy transfer rates of a Förster resonance energy transfer (FRET) pair positioned in close proximity to a 5 nm gold nanoparticle (AuNP) on a DNA origami construct. We study the distance dependence of the FRET rate by varying the location of the donor molecule, D, relative to the AuNP while maintaining a fixed location of the acceptor molecule, A. The presence of the AuNP induces an alteration in the spontaneous emission of the donor (including radiative and non-radiative rates) which is strongly dependent on the distance between the donor and AuNP surface. Simultaneously, the energy transfer rates are enhanced at shorter D-A (and D-AuNP) distances. Overall, in addition to the direct influence of the acceptor and AuNP on the donor decay there is also a significant increase in decay rate not explained by the sum of the two interactions. This leads to enhanced energy transfer between donor and acceptor in the presence of a 5 nm AuNP. We also demonstrate that the transfer rate in the three "particle" geometry (D + A + AuNP) depends approximately linearly on the transfer rate in the donor-AuNP system, suggesting the possibility to control FRET process with electric field induced by 5 nm AuNPs close to the donor fluorophore. It is concluded that DNA origami is a very versatile platform for studying interactions between molecules and plasmonic nanoparticles in general and FRET enhancement in particular

Highly efficient incorporation of the fluorescent nucleotide analogs tC and tCO by Klenow fragment

Studies of the mechanisms by which DNA polymerases select the correct nucleotide frequently employ fluorescently labeled DNA to monitor conformational rearrangements of the polymerase–DNA complex in response to incoming nucleotides. For this purpose, fluorescent base analogs play an increasingly important role because they interfere less with the DNA–protein interaction than do tethered fluorophores. Here we report the incorporation of the 5′-triphosphates of two exceptionally bright cytosine analogs, 1,3-diaza-2-oxo-phenothiazine (tC) and its oxo-homolog, 1,3-diaza-2-oxo-phenoxazine (tCO), into DNA by the Klenow fragment. Both nucleotide analogs are polymerized with slightly higher efficiency opposite guanine than cytosine triphosphate and are shown to bind with nanomolar affinity to the DNA polymerase active site, according to fluorescence anisotropy measurements. Using this method, we perform competitive binding experiments and show that they can be used to determine the dissociation constant of any given natural or unnatural nucleotide. The results demonstrate that the active site of the Klenow fragment is flexible enough to tolerate base pairs that are size-expanded in the major groove. In addition, the possibility to enzymatically polymerize a fluorescent nucleotide with high efficiency complements the tool box of biophysical probes available to study DNA replication

A novel wear-resistant magnetic thin film material based on a Ti1−xFexC1−yTi_{1-x}Fe_xC_{1-y} nanocomposite alloy

In this study we report on the film growth and characterization of thin (approximately 50 nm thick) Ti-Fe-C films deposited on amorphous quartz. The experimental studies have been complemented by first principles density functional theory (DFT) calculations. Upon annealing of as-prepared films, the composition of the metastable Ti-Fe-C film changes. An iron-rich phase is first formed close to the film surface, but with increasing annealing time this phase is gradually displaced toward the film-substrate interface where its position stabilizes. Both the magnetic ordering temperature and the saturation magnetization changes significantly upon annealing. The DFT calculations show that the critical temperature and the magnetic moment both increase with increasing Fe and C-vacancy concentration. The formation of the metastable iron-rich Ti-Fe-C compound is reflected in the strong increase of the magnetic ordering temperature. Eventually, after enough annealing time ($\geq 10$ minutes), nano-crystalline $\alpha$-Fe starts to precipitate and the amount and size of these precipitates can be controlled by the annealing procedure; after 20 minutes of annealing, the experimental results indicate a nano-crystalline iron-film embedded in a wear resistant TiC compound. This conclusion is further supported by transmission electron microscopy studies on epitaxial Ti-Fe-C films deposited on single crystalline MgO substrates where, upon annealing, an iron film embedded in TiC is formed. Our results suggest that annealing of metastable Ti-Fe-C films can be used as an efficient way of creating a wear-resistant magnetic thin film material.Comment: 23 pages, 13 figure

Understanding and protecting vulnerable financial consumers

This article considers how consumer protection law and policy should address the interests of particularly vulnerable financial consumers. Specifically, the article proposes a taxonomy of vulnerability which helps to identify (a) what makes consumers particularly vulnerable, and (b) how consumer protection law and consumer policy (broadly understood) can respond to these causes in a way that provides such consumers with appropriate protection. Changes to economic conditions, legal requirements on traders and our understanding of consumer behaviour make discussion of these issues particularly topical. There is little doubt that finding solutions is extremely difficult. Trade-offs are necessary and some enduring factors that contribute to vulnerability, in particular poverty, sometimes appear intractable. Nevertheless, it is submitted that by identifying clearly both why consumers are vulnerable and how the factors that lead to such vulnerability can be addressed, it is possible to construct an environment which respects consumer choice while ensuring that the most vulnerable are protected appropriately

Tick-transmitted co-infections among erythema migrans patients in a general practice setting in Norway:a clinical and laboratory follow-up study

Background Erythema migrans (EM) is the most common manifestation of Lyme borreliosis. Here, we examined EM patients in Norwegian general practice to find the proportion exposed to tick-transmitted microorganisms other than Borrelia, and the impact of co-infection on the clinical manifestations and disease duration. Methods Skin biopsies from 139/188 EM patients were analyzed using PCR for Neoehrlichia mikurensis, Rickettsia spp., Anaplasma phagocytophilum and Babesia spp. Follow-up sera from 135/188 patients were analyzed for spotted fever group (SFG) Rickettsia, A. phagocytophilum and Babesia microti antibodies, and tested with PCR if positive. Day 0 sera from patients with fever (8/188) or EM duration of ≥ 21 days (69/188) were analyzed, using PCR, for A. phagocytophilum, Rickettsia spp., Babesia spp. and N. mikurensis. Day 14 sera were tested for TBEV IgG. Results We detected no microorganisms in the skin biopsies nor in the sera of patients with fever or prolonged EM duration. Serological signs of exposure against SFG Rickettsia and A. phagocytophilum were detected in 11/135 and 8/135, respectively. Three patients exhibited both SFG Rickettsia and A. phagocytophilum antibodies, albeit negative PCR. No antibodies were detected against B. microti. 2/187 had TBEV antibodies without prior immunization. There was no significant increase in clinical symptoms or disease duration in patients with possible co-infection. Conclusions Co-infection with N. mikurensis, A. phagocytophilum, SFG Rickettsia, Babesia spp. and TBEV is uncommon in Norwegian EM patients. Despite detecting antibodies against SFG Rickettsia and A. phagocytophilum in some patients, no clinical implications could be demonstrated

Potential drivers of human tick-borne encephalitis in the orebro region of Sweden, 2010-2021

Incidence of tick-borne encephalitis (TBE) has increased during the last years in Scandinavia, but the underlying mechanism is not understood. TBE human case data reported between 2010 and 2021 were aggregated into postal codes within orebro County, south-central Sweden, along with tick abundance and environmental data to analyse spatial patterns and identify drivers of TBE. We identified a substantial and continuing increase of TBE incidence in orebro County during the study period. Spatial cluster analyses showed significant hotspots (higher number of cases than expected) in the southern and northern parts of orebro County, whereas a cold spot (lower number of cases than expected) was found in the central part comprising orebro municipality. Generalised linear models showed that the risk of acquiring TBE increased by 12.5% and 72.3% for every percent increase in relative humidity and proportion of wetland forest, respectively, whereas the risk decreased by 52.8% for every degree Celsius increase in annual temperature range. However, models had relatively low goodness of fit (R-2 < 0.27). Results suggest that TBE in orebro County is spatially clustered, however variables used in this study, i.e., climatic variables, forest cover, water, tick abundance, sheep as indicator species, alone do not explain this pattern

A Novel High Discriminatory Protocol for the Detection of Borrelia afzelii, Borrelia burgdorferi Sensu Stricto and Borrelia garinii in Ticks

Bacteria of the Borrelia burgdorferi sensu lato complex are the causative agents of Lyme borreliosis (LB). Even if the conventional diagnosis of LB does not rely on the species itself, an accurate species identification within the complex will provide a deepened epidemiological scenario, a better diagnosis leading to a more targeted therapeutic approach, as well as promote the general public's awareness. A comparative genomics approach based on the 210 Borrelia spp. genomes available in 2019 were used to set up three species-specific PCR protocols, able to detect and provide species typing of Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.) and Borrelia garinii, the three most common and important human pathogenic Lyme Borrelia species in Europe. The species-specificity of these protocols was confirmed on previously identified B. afzelii, B. burgdorferi s.s. and B. garinii specimens detected in Ixodes ricinus samples. In addition, the protocols were validated on 120 DNA samples from ticks collected in Sweden, showing 88% accuracy, 100% precision, 72% sensitivity and 100% specificity. The proposed approach represents an innovative tool in epidemiological studies focused on B. burgdorferi s.l. occurrence in ticks, and future studies could suggest its helpfulness in routine diagnostic tests for health care

Эффективность работы сплит-системы в режиме теплового насоса

Рассмотрены проблемы, возникающие во время работы сплит-системы в режиме теплового насоса и предложена экспериментальная установка, которая даст возможность их исследовать, решить или минимизировать. Разработана методика проведения исследования, обработки полученных данных и расчета показателей эффективности работы сплит-системы в режиме воздушного теплового насоса. Введено понятие цикличности работы сплит-системы и выполнено разделение рабочего цикла на отдельные самостоятельные составляющие. Предложено использовать поправочный коэффициент, который дает возможность получать действительные значения эффективности любого воздушного теплового насоса сплит-системы. Проведена апробация полученных результатов с данными фирм-производителей сплит-систем и выделены особенности, влияющие на значение коэффициента трансформации при использовании разных методов расчета.The problems arising in the operation of the split systems in the thermal pump mode are considered and an experimental unit is proposed which will enable to study, to solve or to minimize them. The methods of the studying, data obtained processing and calculation of the efficiency indices for the split system operation in the mode of the air thermal pump are developed. The notion of cyclicity of the split system operation is introduced and the operation cycle division into separate independent components is provided. It is proposed to use coefficient of correction which enables to obtain actual efficiency values of any air thermal pump of the split system. Testing and comparison of the data obtained with the data of the split system manufacturing companies and their approbation are carried out. The specific features having an influence on the value of transformation ratio when using different methods of calculation are singled out