Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

Use of the Method of Harmonic Balance for Controlling the Condition of Permanent Magnets

Metal Science and Heat Treatment - A method for controlling the condition of permanent magnets based on solution of the problem of harmonic balance and full-scale and model tests is suggested. The...     

Synthesis of Nanoporous Titanosilicates and Their Structural Characterization by Physical Nitrogen Adsorption

Protection of Metals and Physical Chemistry of Surfaces - Mesoporous titanosilicate structures synthesized by the sol–gel method using the cetylpyridinium template at different Ti/Si ratios...     

Assessment of Surface Roughness Bounded by Cosinusoids and Sinusoids

Russian Engineering Research - On the profilograms obtained for real surfaces machined by different methods, the microprojections resemble elements of sinusoids and cosinusoids. Accordingly, it is...     

HA/β-TCP plasma sprayed coatings on Ti substrate for biomedical applications

The present work focuses on the fabrication of βTCP (β-tricalcium phosphate) and HA/βTCP (hydroxyapatite/β-tricalcium phosphate) composite coatings by plasma spraying. The starting powders were produced via solid-state method using 2 wt% MgO to stabilize βTCP phase. The synthesized powders were preliminarily granulated to be used by the plasma spray process. Coatings obtained on titanium substrates are uniform and well adherent but due to the high temperature and cooling rate typical for plasma spraying process, βTCP phase is almost totally transformed into the α allotrope. Thermal treatment at 800 °C allows the reconversion of the phase αTCP→ βTCP. It is therefore possible to produce coatings with tuneable dissolution properties by selecting the proper initial powder mixture and the specific thermal treatment.     

Group sequential crossover trial designs with strong control of the familywise error rate

Crossover designs are an extremely useful tool to investigators, and group sequential methods have proven highly proficient at improving the efficiency of parallel group trials. Yet, group sequential methods and crossover designs have rarely been paired together. One possible explanation for this could be the absence of a formal proof of how to strongly control the familywise error rate in the case when multiple comparisons will be made. Here, we provide this proof, valid for any number of initial experimental treatments and any number of stages, when results are analyzed using a linear mixed model. We then establish formulae for the expected sample size and expected number of observations of such a trial, given any choice of stopping boundaries. Finally, utilizing the four-treatment, four-period TOMADO trial as an example, we demonstrate that group sequential methods in this setting could have reduced the trials expected number of observations under the global null hypothesis by over 33%.     

Application of Mono‐ and Disaccharides in Drug Targeting and Efficacy

Carbohydrates and their conjugates play important roles in many biological processes including fertilization, differentiation, development, immune response, and infection. Their activities are largely dependent on the properties of terminal mono‐ or disaccharides. Galactose, mannose, fucose, glucose, sialic acid, etc., are commonly used as powerful scaffolds installed on drug molecules for targeting specific tissues including brain, liver, and cancers, and as epitopes for enhancing the targeting of various vaccines. This review focuses on the influence of their structural variations, including changes in sugar type, substituent groups and their positions, as well as length of linker portion, on the targeting of drugs or their efficacy. Particular attention is paid to the targeting properties of mono‐ and disaccharides applied in drug design and discovery.     

Carbon Chain Length Modulates MDA-MB-231 Breast Cancer Cell Killing Mechanisms by Mitochondrially Targeted Aryl−Urea Fatty Acids

Targeting the tumor cell mitochondrion could produce novel anticancer agents. We designed an aryl−urea fatty acid ( 1 g ; 16({[4-chloro-3-(trifluoromethyl)phenyl]carbamoyl}amino)hexadecanoic acid) that disrupted the mitochondrion and decreased MDA-MB-231 breast cancer cell viability. To optimize the aryl−ureas the present study evaluated mitochondrial targeting by 1 g analogues containing alkyl chains between 10–17 carbons. Using the dye JC-1, the C12−C17 analogues efficiently disrupted the mitochondrial membrane potential (IC₅₀s 3.5±1.2 to 7.6±1.1 μM) and impaired ATP production; shorter analogues were less active. 7-Aminoactinomycin D/annexin V staining and flow cytometry showed that these agents activated the killing mechanisms of necrosis and apoptosis to varying extents (7-aminoactinomycin D/annexin V staining ratios 4.3–6.0). Indeed, 1 g and its C17 analogue preferentially activated necrosis and apoptosis, respectively (ratios 2.1 and 16). Taken together, alkyl chain length is a determinant of mitochondrial targeting by aryl−ureas and can be varied to develop analogues that activate apoptosis or necrosis in a regulated fashion.