Fiber background rejection and crystal over-response compensation for GaN based in vivo dosimetry

For dosimetric measurements using an implantable optical fiber probe with GaN (Gallium Nitride) scintillator as radioluminescence (RL) transducer, a bi-channel method is proposed to reject the background contribution of the irradiated fiber segment. It is based on spectral differences between the narrow-band light emission from GaN and the large-band background from the irradiated optical fiber. Experimental validation of this method using 6 MV photon beam has shown that the remaining background contribution after subtraction is below 1.2% for square field sizes ranging from 3 cm to 20 cm. Furthermore, a compensation method for the over-response of GaN is also proposed, since GaN is not tissue equivalent. The over-response factor of GaN exhibits a linear increase with square field aperture and depends on depth from phantom surface. This behaviour is modelled to allow compensation in specific conditions. The proposed method has been evaluated and has shown a maximum deviation of 3% for a 6 MV photon beam and 1% for an 18 MV photon beam at a depth beyond the build-up region.     

Synthesis of N-Aminopyrazinium Analogs of Cytidine and 2′-Deoxycytidine

Abstract

N-Aminopyrazine analogues of cytidine and 2′-deoxycytidine were prepared from 1-(β-D-ribofuranosyl)-1,2-dihydro-2-oxopyrazine and 1-(2-deoxy-β-D-ribofuranosyl)-1,2-dihydro-2-oxopyrazine, respectively, by amination with O-mesitylenesulfonylhydroxylamine.     

Activation of Notch Signaling Is Required for Cholangiocarcinoma Progression and Is Enhanced by Inactivation of p53 In Vivo

Cholangiocacinoma (CC) is a cancer disease with rising incidence. Notch signaling has been shown to be deregulated in many cancers. However, the role of this signaling pathway in the carcinogenesis of CC is still not fully explored. In this study, we investigated the effects of Notch inhibition by γ-secretase inhibitor IX (GSI IX) in cultured human CC cell lines and we established a transgenic mouse model with liver specific expression of the intracellular domain of Notch (Notch-ICD) and inactivation of tumor suppressor p53. GSI IX treatment effectively impaired cell proliferation, migration, invasion, epithelial to mesenchymal transition and growth of softagar colonies. In vivo overexpression of Notch-ICD together with an inactivation of p53 significantly increased tumor burden and showed CC characteristics. Conclusion: Our study highlights the importance of Notch signaling in the tumorigenesis of CC and demonstrates that additional inactivation of p53 in vivo.     

Synthesis,Physicochemical Properties,and in vitro Antibacterial Screening of Palladium(II) Complexes Derived from Thiosemicarbazone

A new series of Pd^II complexes derived from thiosemicarbazone has been synthesized. The synthesized Pd^II complexes have been characterized on the basis of elemental analyses, FT‐IR, ¹H‐ and ¹³C‐NMR, UV/VIS, and thermal studies. A square‐planar geometry has been assigned around Pd^II ions on the basis of results obtained from UV/VIS studies. The thiosemicarbazone ligand and its Pd^II complexes have been screened against Gram‐positive (Bacillus subtilis and Staphylococcus aureus) and Gram‐negative (Escherichia coli and Pseudomonas aeruginosa) bacteria in vitro as growth‐inhibiting agents, and the results revealed significant antibacterial activities.     

A metabolomics based approach for understanding the influence of terroir in Vitis Vinifera L.

This study aims to understand the contribution of ‘terroir’ during the cultivation of Vitis vinifera L. The concept of terroir stems from the French ideal that a region’s soil and local vineyard topography together with a region’s macroclimate, including the mesoclimate and vine microclimate, together define the unique characteristics of a wine. In this current study we have utilized high performance liquid chromatography combined with a Q Exactive quadrupole Orbitrap™ mass analyzer for the direct injection analysis of Vitis vinifera juice samples sourced from two different vineyards from the Santa Ynez AVA of Santa Barbara county. Analysis of the mass spectral data was facilitated by a differential analysis software program—SIEVE 2.0™. Distinct metabolomic signatures in freshly crushed juice samples were elucidated. Interestingly, important and distinct information was revealed from the analysis of both the positive and negative ion data. Hierarchical clustering indicated the negative ion data displayed similarity based on varietal character while results obtained in the positive ion mode clustered primarily on terroir. This may indicate that more acidic compounds are influenced by varietal character while more basic compounds are influenced by terroir. Using a feature of SIEVE 2.0 a flavonoid database was utilized to search the raw data for flavonoids present in the juice samples. This targeted analysis indicated the flavonoid profile of juice samples appears to be a good indicator of varietal character independent of terroir. The analysis presented in this study suggests distinct Vitis vinifera grape juice chemical signatures are present prior to fermentation. Further analysis will aim to attribute which of these compounds is influenced by varietal character and/or terroir.

     

The Role of Programmed Cell Death Ligand-1 (PD-L1/CD274) in the Development of Graft versus Host Disease

Programmed cell death ligand-1 (PD-L1/CD274) is an immunomodulatory molecule involved in cancer and complications of bone marrow transplantation, such as graft rejection and graft-versus-host disease. The present study was designed to assess the dynamic expression of this molecule after hematopoietic stem cell transplantation in relation to acute graft-versus-host disease. Female BALB/c mice were conditioned with busulfan and cyclophosphamide and transplanted with either syngeneic or allogeneic (male C57BL/6 mice) bone marrow and splenic cells. The expression of PD-L1 was evaluated at different time points employing qPCR, western blot and immunohistochemistry. Allogeneic- but not syngeneic-transplanted animals exhibited a marked up-regulation of PD-L1 expression in the muscle and kidney, but not the liver, at days 5 and 7 post transplantation. In mice transplanted with allogeneic bone marrow cells, the enhanced expression of PD-L1 was associated with high serum levels of IFNγ and TNFα at corresponding intervals. Our findings demonstrate that PD-L1 is differently induced and expressed after allogeneic transplantation than it is after syngeneic transplantation, and that it is in favor of target rather than non-target organs at the early stages of acute graft-versus-host disease. This is the first study to correlate the dynamics of PD-L1 at the gene-, protein- and activity levels with the early development of acute graft-versus-host disease. Our results suggest that the higher expression of PD-L1 in the muscle and kidney (non-target tissues) plays a protective role in skeletal muscle during acute graft-versus-host disease.     

Is It Possible to Improve Memory Function by Upregulation of the Cholesterol 24S-Hydroxylase (CYP46A1) in the Brain?

We previously described a heterozygous mouse model overexpressing human HA-tagged 24S-hydroxylase (CYP46A1) utilizing a ubiquitous expression vector. In this study, we generated homozygotes of these mice with circulating levels of 24OH 30–60% higher than the heterozygotes. Female homozygous CYP46A1 transgenic mice, aged 15 months, showed an improvement in spatial memory in the Morris water maze test as compared to the wild type mice. The levels of N-Methyl-D-Aspartate receptor 1, phosphorylated-N-Methyl-D-Aspartate receptor 2A, postsynaptic density 95, synapsin-1 and synapthophysin were significantly increased in the hippocampus of the CYP46A1 transgenic mice as compared to the controls. The levels of lanosterol in the brain of the CYP46A1 transgenic mice were significantly increased, consistent with a higher synthesis of cholesterol. Our results are discussed in relation to the hypothesis that the flux in the mevalonate pathway in the brain is of importance in cognitive functions.     

Multi-Host Expression System for Recombinant Production of Challenging Proteins

Recombinant production of complex eukaryotic proteins for structural analyses typically requires a profound screening process to identify suitable constructs for the expression of ample amounts of properly folded protein. Furthermore, the evaluation of an optimal expression host has a major impact on protein yield and quality as well as on actual cost of the production process. Here we present a novel fast expression system for multiple hosts based on a single donor vector termed pFlp-Bac-to-Mam. The range of applications of pFlp-Bac-to-Mam comprises highly efficient transient transfection of HEK293-6E in serum-free suspension culture and subsequent large-scale production of challenging proteins expressing in mg per Liter level using either the baculoviral expression vector system or stable CHO production cell lines generated by Flp-mediated cassette exchange. The success of the multi-host expression vector to identify the optimal expression strategy for efficient production of high quality protein is demonstrated in a comparative expression study of three model proteins representing different protein classes: intracellular expression using a fluorescent protein, secretion of a single-chain-Fv-hIgG1Fc fusion construct and production of a large amount of highly homogeneous protein sample of the extracellular domain of a Toll-like receptor. The evaluation of the production efficiency shows that the pFlp-Bac-to-Mam system allows a fast and individual optimization of the expression strategy for each protein class.