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Fluorescence-based oligonucleotide probes have a great importance in research of molecular interactions. Molecular beacons (MBs) are special case of fluorescent probes that form a stem-loop shape, bringing together a fluorophore and quencher, thus emitting fluorescence only when hybridized to a complementary target. Here we describe a new method for the quantitation of MB hybridization based on the measurement of changes in free energy instead of the fluorescence intensity. The MB energy state can be measured by micro-fluorescence detection. The approach allowed to determine hybridization energy of the MB with target nucleotide directly from fluorescence spectra and distinguish the MB in unfolded and hybridized states. Moreover, the method enabled us to discriminate between DNA duplexes with perfect complementarity or a single-nucleotide mismatch, based on the first direct experimental prove of enthalpy-entropy compensation.

A large number of galaxies have large-scale magnetic fields which are usually measured by the Faraday rotation of radio waves. Their origin is usually connected with the dynamo mechanism which is based on differential rotation of the interstellar medium and alpha-effect characterizing the helicity of the small-scale motions. However, it is necessary to have initial magnetic field which cannot be generated by the dynamo. One of the possible mechanisms is connected with the Biermann battery which acts because of different masses of protons and electrons passing from the central object. They produce circular currents which induce the vertical magnetic field. As for this field we can obtain the integral equation which can be solved by simulated annealing method which is widely used in different branches of mathematics

Coil springs in motorcycles are crucial components in the vehicle's suspension system, where they are used in the front or rear to absorb the impact energy experienced by the wheels. In this study, the coil spring experienced a failure in the form of fracture, prompting several tests to analyze the mechanism and causes of this failure. The conducted tests include visual observation, chemical composition testing, metallographic testing, hardness testing, and FEM simulation. The visual observation results indicate that the failure experienced by the coil spring is due to fatigue fracture, as defined by the presence of crack initiation, crack propagation (beachmark), and final rupture. The chemical composition testing reveals that the material used is SAE 9254 Steel, that is a low-alloy steel. The microstructure formed consisted of ferrite and pearlite phases, attributed to the absence of quenching and tempering processes during the coil spring's production. The hardness testing results show an average hardness of 216,6 HV, confirming the absence of quenching and tempering in the coil spring production process. The FEM simulation results show that the stress does not exceed the yield strength of the material. [ABSTRACT FROM AUTHOR]

In this work, Ce- and Li-doped CaF2 phosphor (CaF2:Ce,Li) was synthesized by the combustion technique. Structural and morphological analysis of the synthesized material has been carried out by X-ray diffraction (XRD) and scanning electron microscopy (SEM), respectively. The XRD pattern reveals the formation of a pure phase of the CaF2 crystal with a crystallite size of 717 nm. The luminescence properties were investigated by the thermoluminescence (TL) technique. The effect of sintering temperature on the TL glow curve of the pellets has been investigated for different temperatures (450, 500, 525, 550, and 575 °C). Pellets sintered at 525 °C and 550 °C presented higher sensitivity in their TL responses for gamma and UV irradiation, respectively. Samples irradiated with gamma radiation show two intense TL peaks centered at 172 °C and 275 °C. On the other hand, samples irradiated with UV radiation show two peaks centered at 195 and 280 °C. The intensity of the TL peaks increases with gamma radiation dose and UV irradiation time. The kinetic parameters of the glow curve peaks were determined through the combined analysis of the results obtained by the Tm-Tstop and deconvolution methods. [ABSTRACT FROM AUTHOR]

Cine-MRI for adhesion detection is a promising novel modality that can help the large group of patients developing pain after abdominal surgery. Few studies into its diagnostic accuracy are available, and none address observer variability. This retrospective study explores the inter- and intra-observer variability, diagnostic accuracy, and the effect of experience. A total of 15 observers with a variety of experience reviewed 61 sagittal cine-MRI slices, placing box annotations with a confidence score at locations suspect for adhesions. Five observers reviewed the slices again one year later. Inter- and intra-observer variability are quantified using Fleiss’ (inter) and Cohen’s (intra) κ and percentage agreement. Diagnostic accuracy is quantified with receiver operating characteristic (ROC) analysis based on a consensus standard. Inter-observer Fleiss’ κ values range from 0.04 to 0.34, showing poor to fair agreement. High general and cine-MRI experience led to significantly (p < 0.001) better agreement among observers. The intra-observer results show Cohen’s κ values between 0.37 and 0.53 for all observers, except one with a low κ of −0.11. Group AUC scores lie between 0.66 and 0.72, with individual observers reaching 0.78. This study confirms that cine-MRI can diagnose adhesions, with respect to a radiologist consensus panel and shows that experience improves reading cine-MRI. Observers without specific experience adapt to this modality quickly after a short online tutorial. Observer agreement is fair at best and area under the receiver operating characteristic curve (AUC) scores leave room for improvement. Consistently interpreting this novel modality needs further research, for instance, by developing reporting guidelines or artificial intelligence-based methods.

Fluorescence-based oligonucleotide probes have a great importance in research of molecular interactions. Molecular beacons (MBs) are special case of fluorescent probes that form a stem-loop shape, bringing together a fluorophore and quencher, thus emitting fluorescence only when hybridized to a complementary target. Here we describe a new method for the quantitation of MB hybridization based on the measurement of changes in free energy instead of the fluorescence intensity. The MB energy state can be measured by micro-fluorescence detection. The approach allowed to determine hybridization energy of the MB with target nucleotide directly from fluorescence spectra and distinguish the MB in unfolded and hybridized states. Moreover, the method enabled us to discriminate between DNA duplexes with perfect complementarity or a single-nucleotide mismatch, based on the first direct experimental prove of enthalpy-entropy compensation. [ABSTRACT FROM AUTHOR]

Nucleobase building blocks have been demonstrated to be strong candidates when it comes to DNA/RNA-like materials by benefiting from hydrogen bond interactions as physical properties. Modifying at the 5′ position is the simplest way to develop nucleobase-based structures by transesterification using the lipase Novozym 435. Herein, we describe the optimization of the lipase-catalyzed synthesis of the monomer 5′-O-methacryloylcytidine with the assistance of microwave irradiation. Variable reaction parameters, such as enzyme concentration, molar ratio of the substrate, reaction temperature and reaction time, were investigated to find the optimum reaction condition in terms of obtaining the highest yield.

La miocardiopatía o displasia arritmogénica del ventrículo derecho es una cardiopatía de origen genético cuyo diagnóstico supone, a menudo, un reto para el clínico. Es una de las causas más comunes de muerte súbita cardíaca en la adolescencia y en los adultos jóvenes. Se presenta el caso de un paciente con historia de arritmias ventriculares malignas y de muerte súbita cardíaca recuperada, por displasia arritmogénica del ventrículo derecho, con fragmentación del QRS en las derivaciones precordiales derechas, como marcador de la presencia de un sustrato propicio para el surgimiento de la fibrilación ventricular espontánea. Se comenta la patogenia, el diagnóstico y el tratamiento de esta enfermedad.

Abstract Chimeric antigen receptor (CAR) T cells brought substantial benefit to patients with B‐cell malignancies. Notwithstanding, CAR T‐cell manufacturing requires complex procedures impeding the broad supply chain. Here, we provide evidence that human CD19‐CAR T cells can be generated directly in vivo using the lentiviral vector CD8‐LV specifically targeting human CD8+ cells. Administration into mice xenografted with Raji lymphoma cells and human peripheral blood mononuclear cells led to CAR expression solely in CD8+ T cells and efficacious elimination of CD19+ B cells. Further, upon injection of CD8‐LV into mice transplanted with human CD34+ cells, induction of CAR T cells and CD19+ B‐cell depletion was observed in 7 out of 10 treated animals. Notably, three mice showed elevated levels of human cytokines in plasma. Tissue‐invading CAR T cells and complete elimination of the B‐lymphocyte‐rich zones in spleen were indicative of a cytokine release syndrome. Our data demonstrate the feasibility of in vivo reprogramming of human CD8+ CAR T cells active against CD19+ cells, yet with similar adverse effects currently notorious in the clinical practice.

A spectrum of immune states resulting from tumor resident macrophages and T-lymphocytes in the solid tumor microenvironment correlates with patient outcomes. We hypothesized that in gastric cancer (GC), macrophages in a polarized immunosuppressive transcriptional state would be prognostic of poor survival. We derived transcriptomic signatures for M2 (M2TS, MRC1; MS4A4A; CD36; CCL13; CCL18; CCL23; SLC38A6; FGL2; FN1; MAF) and M1 (M1TS, CCR7; IL2RA; CXCL11; CCL19; CXCL10; PLA1A; PTX3) macrophages, and cytolytic T-lymphocytes (CTLTS, GZMA; GZMB; GZMH; GZMM; PRF1). Primary GC in a TCGA stomach cancer dataset was evaluated for signature expressions, and a log-rank test determined overall survival (OS) and the disease-free interval (DFI). In 341 TCGA GC entries, high M2TS expression was associated with histological types and later stages. Low M2TS expression was associated with significantly better 5-year OS and DFI. We validated M2TS in prospectively collected peritoneal fluid of a GC patient cohort (n = 28). Single-cell RNA sequencing was used for signature expression in CD68+CD163+ cells and the log-rank test compared OS. GC patients with high M2TS in CD68+CD163+ cells in their peritoneal fluid had significantly worse OS than those with low expression. Multivariate analyses confirmed M2TS was significantly and independently associated with survival. As an independent predictor of poor survival, M2TS may be prognostic in primary tumors and peritoneal fluid of GC patients. [ABSTRACT FROM AUTHOR]

Glycans carry a vast range of functions in nature. Utilizing their properties and functions in form of polymers, coatings or glycan derivatives for various applications makes the synthesis of modified glycans crucial. Since amines are easy to modify for subsequent reactions, we investigated regioselective amination conditions of different saccharides. Amination reactions were performed according to Kochetkov and Likhoshertov and accelerated by microwave irradiation. We optimized the synthesis of glycosylamines for N-acetyl-d-galactosamine, d-lactose, d-glucuronic acid and l-(−)-fucose using the design of experiments (DoE) approach. DoE enables efficient optimization with limited number of experimental data. A DoE software generated a set of experiments where reaction temperature, concentration of carbohydrate, nature of aminating agent and solvent were investigated. We found that the synthesis of glycosylamines significantly depends on the nature of the carbohydrate and on the reaction temperature. There is strong indication that high temperatures are favored for the amination reaction.

[This corrects the article DOI: 10.1371/journal.pone.0018823.].

Hepatocellular carcinoma (HCC) is frequently detected in pre-existing liver cirrhosis, but can also develop without such pre-conditions. There is an increasing trend of HCC incidence worldwide. In patients with liver cirrhosis, HCC has become the leading cause of death. At diagnosis the tumor has very often reached an advanced stage and curative treatment options are missing. Thus, early diagnosis would help the patient and prevent increasing healthcare costs. In our review we will summarize the recommendations of the German S3 guideline for the early diagnosis of HCC and will discuss the current literature in this context. The reader will learn which diagnostic tools are available and in what order they can be usefully applied. Surveillance should be done with ultrasound by a skilled examiner, additional imaging at best with state-of-the-art dynamic magnetic resonance.

The present study analyses the transmission of vibrations generated from a multidirectional trunk shaker to olive tree structure considering both the aerial zone (trunk and branches) and the underground zone (the coarse root). The vibration characterization was conducted by measuring acceleration on several points of the tree during harvesting operations. The influence of two different heights of shaker head clamping was analysed. In addition, a dynamic probing was performed in order to evaluate soil compaction. The results showed that the vibration performed by the trunk shaker head, corresponding to an acceleration resultant of approximately 77 ms–2 with a dominant vibration frequency of 18 Hz, increased up to 106% in branches and decreased up to 90% in trunks. At root level, where the analysis was carried out at 1/3 and 2/3 of the coarse root length, the acceleration values diminished significantly to 17 ms–2 and 12 ms–2, respectively. Soil dynamic resistance was lower (36 kg cm–2) near the trees than between the trees (53 kg cm–2). The vibration transmission to the aerial and the underground parts diversely influences the dynamic behaviour of the olive tree, considering an operational frequency of a commercial trunk shaker. The assessment of vibration transmission to the aerial part could contribute to improve fruit detachment and reduce branch breaking and leaf detachment. While vibration transmission to the underground part rises new challenges considering soil compaction in olive groves.

[This corrects the article DOI: 10.1371/journal.pone.0077433.].

Intensive mining and processing of the polymetallic sulfide ore body of Cerro Rico de Potosí (Bolivia) has occurred since 1545. To further investigate acid mine drainage (AMD) discharges and their link to downstream contamination, data were gathered during two sampling events during the most extreme periods of the dry and wet seasons of one year. Concentrations of Ag, B, Ba, Mo, Sb, Se, Sn and V in AMD and receiving streams were greater than Bolivian discharge limits and receiving water body guidelines as well as international agricultural use standards. High concentrations of rare earth metals have been documented in this area. Results indicate that contamination from mining has a larger scope than previously thought and underscore the importance of remediation.