High-resolution laser ablation inductively coupled plasma mass spectrometry used to study transport of metallic nanoparticles through collagen-rich microstructures in fibroblast multicellular spheroids

We have efficiently produced collagen-rich microstructures in fibroblast multicellular spheroids (MCSs) as a three-dimensional in vitro tissue analog to investigate silver (Ag) nanoparticle (NP) penetration. The MCS production was examined by changing the seeding cell number (500 to 40,000 cells) and the growth period (1 to 10 days). MCSs were incubated with Ag NP suspensions with a concentration of 5 μg mL⁻¹ for 24 h. For this study, laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used to visualize Ag NP localization quantitatively. Thin sections of MCSs were analyzed by LA-ICP-MS with a laser spot size of 8 μm to image distributions of ¹⁰⁹Ag, ³¹P, ⁶³Cu, ⁶⁶Zn, and ⁷⁹Br. A calibration using a NP suspension was applied to convert the measured Ag intensity into the number of NPs present. The determined numbers of NPs ranged from 30 to 7200 particles in an outer rim of MCS. The particle distribution was clearly correlated with the presence of ³¹P and ⁶⁶Zn and was localized in the outer rim of proliferating cells with a width that was equal to about twice the diameter of single cells. Moreover, abundant collagens were found in the outer rim of MCSs. For only the highest seeding cell number, NPs were completely captured at the outer rim, in a natural barrier reducing particle transport, whereas Eosin (⁷⁹Br) used as a probe of small molecules penetrated into the core of MCSs already after 1 min of exposure.

Fibroblast MCS could build up the barrier only for nanoparticles

     

De Novo Designed α-Helical Coiled-Coil Peptides as Scaffolds for Chemical Reactions

Coiled coils (CCs) are well-understood protein-folding motifs. They appear in a variety of oligomer states and as homo- and heteromeric assemblies. This versatility and the general accessibility by de novo design makes them ideal building blocks for synthetic biology. This Minireview highlights the efforts being made in designing small peptide catalysts or reaction templates based on the CC scaffold. The first reports described autocatalysis or mediation of peptide ligation based on CC recognition. Over the years, the designs became more advanced, catalyzing ester hydrolysis, acyl transfer and redox reactions with partial enzyme-like reactivity. Due to the ability to control CC assembly, and, in heterodimeric systems, the association and dissociation, the CC motif has become a common peptide tag in chemical biology.     

An optically stimulated luminescence study of flint related to radiation dosimetry

This article describes the essential features of photo-stimulated luminescence in flint. The emission is shown to be complex, arising from a number of different aspectsof the material. Under constant illumination, dose-dependent time-decaying anti-Stokes signals are superimposed on dose-independent, time-stable emission that is both Stokes and anti-Stokes shifted in energy. It is argued that the dose-dependent signals arise from the crystalline quartz phase of the material, whereas the dose independent signals originate from an amorphous-like phase of SiO₂. The microcrystallinity of the material leads to contrasting behaviour when compared with bulk crystalline quartz.     

Experimental determinations of beta attenuation in planar dose geometry and application to ESR dating of tooth enamel

Two experiments were performed to determine the distribution of beta dose in a target medium as a function of distance from a planar source (2π geometry). In Experiment 1 planar absorbers were used to simulate depth in the target medium, and the attenuated dose that penetrated the intervening absorbers was detected using thermoluminescence from CaSO₄:Tm. In experiment 2 pellets of powdered tooth enamel were situated in a cavity in solid tooth enamel. Dose response of the electron spin resonance (ESR) signals in the pellets allowed direct detection of dose distribution within the target medium. The dose distributions were found to agree more closely with Monte Carlo calculations and with one-group transport theory (Prestwich et al., 1997 and Brennan et al., 1997) than with approximations used widely in ESR dating applications (Grün, 1986). Although the latter were supported by experiments by Aitken et al. (1985), the results of experiment 1 provided data to suggest that departure from 2π geometry was unacceptably large in the Aitken et al. (1985) configuration. The beta doses obtained by one-group theory in typical target volumes of enamel were 17 to 40% lower than those obtained using the Grün approximation. It was concluded that one-group transport theory provides the best available routine tool for calculations of beta dose attenuation in near-planar target materials (e.g. tooth enamel and shell) irradiated by natural sources, although Monte Carlo methods are better and could eventually be introduced into the ROSY ESR dating program (Brennan et al., 1997). It was also concluded that a revision of previously published ESR ages using one-group transport for beta dosimetry will be needed. For cases where the U concentrations in enamel are generally low (less than a few ppm), previously published ESR ages on tooth enamel are systematically too young by 5–30%.     

A new combination of protecting groups and links for encoded synthetic libraries suited for consecutive tests on the solid phase and in solution

Summary A strategy for high-throughput evaluation of combinatorial compound libraries is reported, which circumvents the necessity to test complex mixtures. The method is based on a new combination of protecting groups, solid-phase linker and tags. The bulk of the library first undergoes a binding assay with the components grafted on beads. A selection of beads carrying strong ligands is stripped from the labelled target and distributed into microvessels. The ligands are cleaved and rinsed into microeluates. Subsequently, a more detailed characterization with a functional assay in solution determines the best performers, which are identified through the peptidic tag left behind on the corresponding mother bead.Abbreviations Boc tert-butyloxycarbonyl - CCL combinatorial compound libraries - Ddz ,-dimethyl-3,5-dimethoxybenzyloxycarbonyl - DICD N,N-diisopropylcarbodiimide - DIPEA diisopropylethylamine - DMA dimethylacetamide - ESL encoded synthetic libraries - FITC fluoresceinisothiocyanate - GABA -aminobutyric acid - HOBT N-hydroxybenzotriazole - MALDI matrix-assisted laser desorption ionization - Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl - Pmc 2,2,5,7,8-pentamethylchroman-6-sulfonyl - TFA trifluoroacetic acid - Trt trityl     

On the preparation of 4-mercapto-azetidinones

4-Dithioazetidinone and 4.7-diaza-2-thiabicyclo[3.2.0]hept-2-en-6-one derivatives were prepared from penam-3-isocyanates, which are useful intermediates to 4-mercapto-azetidinones.     

Synthesis, modeling, and anti-tubulin activity of a D-seco paclitaxel analogue

[reaction: see text] We have previously described a model of paclitaxel-microtubule binding that led to the prediction that analogues of paclitaxel lacking any D ring could stabilize microtubules as well as paclitaxel if the substituent present at C4 did not have unfavorable steric interactions with the binding pocket. We report the synthesis of a 4-methyl paclitaxel analogue, compound 1, which bears this prediction out. Compound 1 is as potent as paclitaxel at microtubule stabilization in vitro; however, it has only about one-four-hundredth the cytotoxicity of paclitaxel.     

A systematic SAR study of C10 modified paclitaxel analogues using a combinatorial approach

A library with 63 paclitaxel analogues modified at the C10 position of paclitaxel has been prepared using parallel solution phase synthesis. Most of the C10 analogues were slightly less active than paclitaxel in the tubulin assembly assay and had reduced potency in the B16 melanoma and MCF-7 cell line cytotoxicity assays. These modifications at C10, however, did not lead to the total loss of activity, indicating that the C10 moiety of paclitaxel may not be directly involved in the drug-microtubule interactions, but could influence its binding affinity to P-glycoprotein. Approximately 50% of the analogues demonstrated better activity against the drug resistant cell line MCF7-ADR. However, the increase in activity was 10-fold at most. This result demonstrates that the cytotoxicity against this drug resistant cancer cell line is sensitive to structural changes at the C10 position of paclitaxel. It was also found that the presence of a nitrogen atom in the C10 substituent might play a role in the interaction of analogues with microtubules.     

Mannich Reactions of Cephalosporin Sulphoxides and Sulphones with Imonium Salts. An Improved Synthesis of 2-Methylene-cephalosporins

2-Methylenecephem-1-oxides (4) or -1,1-dioxides (6) can be very conveniently obtained through the use of dimethyl(methylene)imonium salts, which can be prepared in situ or in advance from bis(N,N-dimethylamino)methane (2) and a reactive acid derivative.     

A study of the yield and post‐yield behavior of syndiotactic polypropylene

The strain recovery of three syndiotactic polypropylenes (s‐PPs) differing in the percentage of [rrrr] pentad is investigated. A suitable method based on loading–unloading tests at constant displacement rate in tensile loading conditions is adopted to measure the residual and recovered strain components of the applied strain. The method allows to obtain a large amount of data from few tests and to explore a wide strain range. The dependence of the material's strain recovery on the applied strain is analyzed in relation to s‐PP strain‐induced microstructural changes and crystalline form transitions, which are reported in literature. © 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2014 , 52, 1276–1282