Synthesis of polymeric microspheres from a Merrifield resin by surface‐initiated nitroxide‐mediated radical polymerization

Polymeric microspheres were prepared from a Merrifield resin via nitroxide‐mediated radical polymerization. Polystyrene, poly(acetoxystyrene), and poly[styrene‐b‐(methyl methacrylate‐co‐styrene)], poly(acetoxystyrene‐b‐styrene), and poly(styrene‐co‐2‐hydroxyethyl methacrylate) copolymers were demonstrated to graft onto 2,2,6,6‐tetramethyl‐1‐piperidinyloxy nitroxide bound Merrifield resins. The polymerization control was enhanced both on the surface and in solution by the addition of sacrificial nitroxide. The significant increase in the particle diameter (more than a fivefold volume increase for polystyrene brushes) showed that polymer growth was not only on the surface but also within the particles, and this diameter increase could be adjusted through changes in the molecular weight of the polymers. The microspheres were characterized by elemental analysis, IR spectroscopy, particle size analysis, and optical microscopy. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 2145–2154, 2005     

Nitroxide‐mediated homo‐ and block copolymerization of styrene and multifunctional acryl‐ and methacryl derivatives

The ability of different alkoxyamines ( I1 , I2 , I3 , I4 , and I5 ) to initiate controlled radical polymerization of styrene was evaluated. Among them, 2‐hydroxymethyl‐2‐[(2‐methyl‐1‐phenyl‐propyl)‐(1‐phenyl‐ethoxy)‐amino]‐propane‐1,3‐diol ( I5 ) gave the highest polymerization rate of styrene, and the best control over the molecular weight and the molecular weight distribution of polystyrene. Kinetic studies confirmed that with initiator I5 the polymerization of styrene proceeded in a controlled way. The controlled radical homopolymerization of multifunctional acryl‐ and methacryl derivatives using initiator I5 could not be realized as demonstrated by the high polydispersities (PD) obtained. However, it was possible to polymerize multifunctional acryl‐ and methacryl derivatives using a polystyrene macroinitiator ( Pst ) and, thus, novel amphiphilic block copolymers with a narrow molecular weight distribution were obtained. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 1873–1882, 2005     

Beiträge zur Chemie der Pyrrolpigmente, 80. Mitt. Synthese,Struktur und Transporteigenschaften von Hexapyrrinanaloga

Summary A 15-thia- and a 15-nor hexapyrrin derivative were prepared. Their structural features were analyzed by¹H-NMR and absorption spectroscopy. All-(Z) configurations and all-syn conformations were found throughout. Carrier mediated transport efficiencies of these compounds in bulk membranes were studied and compared with those of a recently synthesized pentapyrrin-dione and their tripyrrinone partial structure. The pentapyrrin-dione derivative turned out to be one of the most efficient Hg(II) carriers known so far.

     

Beiträge zur Chemie der Pyrrolpigmente, 47. Mitt.

Carrier mediated transport of bilirubin, biliverdin, their dimethylesters and aetiobiliverdin-IV- is measured using a variety of potential carriers like proteins, peptides, tensides and -cyclodextrin in a bulk membrane model. A strong dependence of the transport flux densities on structural details of the carriers as well as of the bile pigments themselves is observed.

     

Mathematical model to reduce loop mediated isothermal amplification (LAMP) false‐positive diagnosis

Loop mediated isothermal amplification (LAMP) is a nucleic acid amplification technique performed under isothermal conditions. The output of this amplification technique includes multiple different sizes of deoxyribonucleic acid (DNA) structures which are identified by a banding pattern on gel electrophoresis plots. Although this is a specific amplification technique, the complexity of the primer design and amplification still lead to the issue of obtaining false‐positive results, especially when a positive reading is determined solely by whether there is any banding pattern in the gel electrophoresis plot. Here, we first performed extensive LAMP experiments and evaluated the DNA structures using microchip electrophoresis. We then developed a mathematical model derived from the various components that make up an entire LAMP structure to predict the full LAMP structure size in base pairs. This model can be implemented by users to make predictions for specific, DNA size dependent, banding patterns on their gel electrophoresis plots. Each prediction is specific to the target sequence and primers used and therefore reduces incorrect diagnosis errors through identifying true‐positive and false‐positive results. This model was accurately tested with multiple primer sets in house and was also translatable to different DNA and RNA types in previously published literature. The mathematical model can ultimately be used to reduce false‐positive LAMP diagnosis errors for applications ranging from tuberculosis diagnostics to E. coli to numerous other infectious diseases.     

Synthesis of asymmetrically substituted head‐to‐head polyacetylenes from 2,3‐disubstituted‐1,3‐butadienes

Asymmetrically substituted head‐to‐head polyacetylenes with phenyl and triphenylamine, thienyl or pyrenyl side groups were synthesized through anionic or controlled radical polymerization of 2,3‐disubstituted‐1,3‐butadienes and subsequent dehydrogenation process. Anionic polymerizations of the designed monomers bearing pendent triphenylamine and thienyl group gave narrow disperse disubstituted precursor polybutadienes with exclusive 1,4‐ or 4,1‐structure, which were confirmed by GPC and NMR measurements. In addition, the monomers possessing pyrenyl group were polymerized via nitroxide mediated radical polymerization and the resulting polymers were obtained with controlled molecular weight and low polydispersities. These polybutadiene precursors were then dehydrogenated in the presence of 2,3‐dichloro‐5,6‐dicyano‐1,4‐benzoquinone. Thus asymmetrically substituted head‐to‐head polyacetylenes were obtained as indicated by ¹H NMR. The properties of polybutadiene precursors and the corresponding polyacetylenes were analyzed by UV–vis, DSC, and TGA. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2019 , 57, 395–402     

Computer simulation of the enantioselective separation of weak bases in an online capillary electrophoresis based microanalysis configuration comprising sulfated cyclodextrin as selector

Computer simulation was utilized to characterize the electrophoretic processes occurring after reactant mixing in an online assay format used for monitoring the enantioselective N‐demethylation of ketamine to norketamine in the presence of highly sulfated γ‐cyclodextrin (HS‐γ‐CD). The incubated reaction mixture (at pH 7.4 and without chiral selector) is bracketed by a low pH BGE containing 2% HS‐γ‐CD as chiral selector, thereby forming a discontinuous buffer system. Upon power application, simulation provides insight into the formation of moving boundaries and new zones together with the prediction of the behavior of ketamine and norketamine enantiomers. The analytes first migrate cationically in a zone electrophoretic manner until they come in contact with HS‐γ‐CD upon which enantioseparation is initiated. Complexation has a focusing effect and the electrophoretic transport becomes reversed, that is, toward the anode. Simulation revealed that the initial conditions for the chiral separation, including buffer components concentrations, pH, and ionic strength, are different than those in the BGE. As a consequence thereof, the experimentally determined complexation parameters for the BGE were unable to correctly describe the migration behavior of the analytes in this column section. An increase in the input binding constants by a factor of two to four, as a result of the decreased ionic strength, resulted in simulation data that agreed with experimental findings.     

Glycosylated Cell‐Penetrating Poly(disulfide)s: Multifunctional Cellular Uptake at High Solubility

The glycosylation of cell‐penetrating poly(disulfide)s (CPDs) is introduced to increase the solubility of classical CPDs and to achieve multifunctional cellular uptake. With the recently developed sidechain engineering, CPDs decorated with α‐d ‐glucose (Glu), β‐d ‐galactose (Gal), d ‐trehalose (Tre), and triethyleneglycol (TEG) were readily accessible. Confocal laser scanning microscopy images of HeLa Kyoto cells incubated with the new CPDs at 2.5 μm revealed efficient uptake into cytosol and nucleoli of all glycosylated CPDs, whereas the original CPDs and TEGylated CPDs showed much precipitation into fluorescent aggregates at these high concentrations. Flow cytometry analysis identified Glu‐CPDs as most active, closely followed by Gal‐CPDs and Tre‐CPDs, and all clearly more active than non‐glycosylated CPDs. In the MTT assay, all glyco‐CPDs were non‐toxic at concentrations as high as 2.5 μm . Consistent with thiol‐mediated uptake, glycosylated CPDs remained dependent on thiols on the cell surface for dynamic covalent exchange, their removal with Ellman's reagent DTNB efficiently inhibited uptake. Multifunctionality was demonstrated by inhibition of Glu‐CPDs with d ‐glucose (IC₅₀ ca. 20 mm ). Insensitivity toward l ‐glucose and d ‐galactose and insensitivity of conventional CPDs toward d ‐glucose supported that glucose‐mediated uptake of the multifunctional Glu‐CPDs involves selective recognition by glucose receptors at the cell surface. Weaker but significant sensitivity of Gal‐CPDs toward d ‐galactose but not d ‐glucose was noted (IC₅₀ ca. 110 mm ). Biotinylation of Glu‐CPDs resulted in the efficient delivery of streptavidin together with a fluorescent model substrate. Protein delivery with Glu‐CPDs was more efficient than with conventional CPDs and remained sensitive to DTNB and d ‐glucose, i.e., multifunctional.     

Hydrogen bonding cooperativity of water to nucleotide recognition: structure of octadecahydrated inosine 5′-monophosphate, 2(C10H12N4O8P)·18H2O at atomic resolution

The crystal structure of an octadecahydrated complex between two inosine 5-monophosphate (IMP) has been determined at atomic resolution, which reveals the hydrogen bonding and the coordination cooperativity of water molecules to nucleotide recognition. The crystal belongs to monoclinic space group P2₁ with cell dimensions a = 8.65(1), b = 21.90(1), c = 12.37(1)Å, and = 110.38(9)°. The ribose hydroxyls, purine N7, keto(O6) bonded water molecules W1, W2, W5, W6, W8 and the phosphate bridge forming water oxygens of W4, W7, W11 appear to play an invariant role in their hydrogen bonding interactions with the IMPs. The synergistic role of the water molecules W5, W6, W8 in the purine staking domain N27W5=2.583,O16W8=2.759,O2627W6=2.723 Åhave been clearly observed for the first time. The complexation of the water molecules through variable hydrogen bonding coordination indicate their functional involvement through extensive cooperative donor-acceptor network mechanism. The occurrence of hydrogen-bonded water spines, water bridges and their interplay in the structural association of IMPs could indicate the possible viability of those aquatic centers in the biological situation.     

Improved adapter-ligation-mediated allele-specific amplification for multiplex genotyping by using software

Adapter-ligation-mediated allele-specific amplification (ALM-ASA) is a potential method for multiplex SNPs typing at an ultra low cost. Here, we describe a kind of software, which designs allele-specific primers for ALM-ASA assay on multiplex SNPs. DNA sequences containing SNPs of interest are submitted into the software which contains various endonucleases for options. Based on the SNP sequence information and the selected endonucleases, the software is capable of automatically generating sets of information needed to perform genotyping experiments. Each set contains a suitable endonuclease, qualified allele-specific primers with orientations and melting temperatures, sizes of allele-specific amplicons, and gel electropherograms simulated according to the sizes of the allele-specific amplicons and the mobility of DNA fragments in 2% agarose gel. Seven SNPs in the arylamines N-acetyltransferase 2 (NAT2) gene, five SNPs in the BRCA1 gene, five SNPs in the COMT gene, six SNPs in the CYP2E1 gene, five SNPs in the MPO gene, and six SNPs in the NRG1 gene were selected for evaluating the software. Without extra optimization, seven SNPs in the NAT2 gene were successfully genotyped for genomic DNA samples from 127 individuals by using the first set of allele-specific primers yielded by the software. Although several steps are used in the ALM-ASA assay, the whole genotyping process can be completed within 3 h by optimizing each step. Profiting from the software, the ALM-ASA assay is easy-to-perform, labor-saving, and accurate.