Synthesis and Biodegradability of Tartaric Acid-Based Poly(ester-thioether)s via Thiol-Ene Click Polymerization.

Using scandium triflate [Sc(OTf)3] as a catalyst, chemoselective esterification of tartaric acids by 3-butene-1-ol was performed, and we produced three dialkene monomers: l-di(3-butenyl) tartrate (BTA), d-BTA, and meso-BTA. Thiol-ene polyaddition of these dialkenyl tartrates and dithiols including 1,2-ethanedithiol (ED), ethylene bis(thioglycolate) (EBTG), and d,l-dithiothreitol (DTT) proceeded in toluene at 70 °C under nitrogen to give tartrate-containing poly(ester-thioether)s (Mn, (4.2-9.0) × 103; molecular weight distribution (Mw/Mn), 1.6-2.5). In differential scanning calorimetry, the poly(ester-thioether)s showed single Tgs between -25 and -8 °C. In biochemical oxygen demand (BOD) tests using activated sludge, poly(l-BTA-alt-EBTG) and poly(l-BTA-alt-ED) showed 32 and 8% biodegradability, which is comparable to that of similar l-malate-containing poly(ester-thioether)s (23 and 13% biodegradation, respectively). Notably, we observed enantio and diastereo effects on biodegradation because poly(l-BTA-alt-EBTG), poly(d-BTA-alt-EBTG), and poly(meso-BTA-alt- EBTG) showed different degradation behaviors during the biodegradation test (BOD/theoretical oxygen demand (TOD) values after 28 days, 32, 70, and 43%, respectively). Our findings provide insights into the design of biomass-based biodegradable polymers containing chiral centers.

Catenane formation of a cyclic poly(alkyl sorbate) via chain-growth polymerization induced by an N-heterocyclic carbene and ring-closing without extreme dilution.

1,3-Di-tert-butylimidazol-2-ylidene (NHCtBu), a typical N-heterocyclic carbene (NHC), was previously found to induce the anionic chain-growth polymerization of ethyl sorbate (ES) in the presence of an aluminum Lewis acid, i.e., methylaluminum bis(2,6-di-tert-butyl-4-methylphenoxide) (MAD), in which the neighboring of α-terminal dienolate with a propagating anion induced cyclization without highly diluted conditions, after monomer depletion, to give the cyclic poly(ES). In this paper, we report that catenane formation occurs by two-step polymerization of ethyl sorbate (ES), in which, after complete monomer (ES) consumption ([ES]0/[NHCtBu]0 = 100/1) in toluene followed by purification by reprecipitation, a second addition of ES monomer ([ES]0/[ NHCtBu]0 = 20/1) in another pot (in toluene or tetrahydrofuran (THF)) resulted in catenane formation, namely a polycatenane. TEM images of a sample from the second step polymerization in THF revealed particles of polycatenane structure consisting of cyclic poly(ES) with sizes ranging from 200 to 1000 nm, showing that this NHCtBu triggered chain polymerization and successive cyclization without highly diluted conditions enabled us to fabricate the intended polycatenane in the successive two-step polymerization.

N-Heterocyclic Carbene Initiated Anionic Polymerization of (E,E)-Methyl Sorbate and Subsequent Ring-Closing to Cyclic Poly(alkyl sorbate).

A diene-based cyclic polymer has been synthesized by the anionic polymerization of methyl sorbate (MS) by an N-heterocyclic carbene (NHC) in the presence of a bulky aluminum Lewis acid. We first polymerized methyl sorbate (MS) initiated by NHC in N,N-dimethylformamide (DMF) at 25 °C, poly(MS) with a number-average molecular weight (Mn) of 3.5 × 103 (Mw/Mn = 2.1) was obtained with a conversion of 93%. The structure was confirmed by 1H and 13C NMR and IR spectra, which revealed that the propagation proceeded via 1,2-addition as well as 1,4-addition. Although the polymerization did not occur in toluene in the absence of any additive, quantitative monomer consumption was observed in the presence of methylaluminum bis(2,6-di-tert-butyl-4-methylphenoxide) (MAD) to afford the poly(MS) with a 1,4-trans structure, 86% of threo diastereoselectivity, and a Mn of 23.0 × 103 with narrow molecular weight distribution (Mw/Mn = 1.17). From the matrix assisted laser desorption/ionization (MALDI-TOF) mass spectra of poly(MS) and the hydrogenated analogue, ring-closing occurred by nucleophilic attack of the anionic propagating center into the adjacent carbon of the α-terminal imidazolimium group to afford cyclic poly(MS). The cyclic formation in the present synthesis system was confirmed by DSC and viscosity measurements.

Synthesis of Glycopolymer Containing Cell-Penetrating Peptides as Inducers of Recombinant Protein Expression under the Control of Lactose Operator/Repressor Systems.

We recently reported on newly synthesized S-galactosyl oligo(Arg) conjugates to overcome the serious problem of the passage through the E. coli cell membrane. Following in vivo expression of green fluorescent protein (GFP) induced by each of the S-galactosyl (Arg)n constructs (n = 5, 6, 8) at the T5 promoter in E. coli for 18 h, we visually observed that the cultures fluoresced green light when excited with UV light. The fluorescence intensities for these cultures were greater than that found for a control culture, indicating that the peptides had induced GFP expression. In order to accomplish higher expression efficiency, we investigated the cluster effect and structural fine-tuning of new poly(2-oxazoline) containing CysArgArg as the cell-penetrating peptide (CPP) and S-galactosides when acting as inducers of recombinant protein expression under the control of lac operator/repressor systems in this article. Quantitative fluorescence intensities (calculated per molecule) also supported the observations that the cell-penetrating glyco poly(2-oxazoline)s were better inducers of GFP expression than glyco poly(2-oxazoline) containing no CPP or isopropyl ß-d-thiogalactoside. Because the level of GFP expression was directly related to the number of sugar residues in each glyco poly(2-oxazoline), we propose that a cluster effect of the S-galactosides attached to the cell-penetrating poly(2-oxazoline) is responsible for how well the galactosides inhibited the lac repressor to activate the protein expression under the control of the lac operator/repressor system. A similar tendency was observed when the T7 promoter was placed upstream of the gene for an artificial extracellular matrix protein and glyco poly(2-oxazoline)s-CPP conjugates were used as inducers. To assess how the glyco poly(2-oxazoline) penetrate the cell membrane, we labeled the glyco poly(2-oxazoline) using 1-amino pyrene and directly observed the penetration process. Furthermore, we could visualize protein expression under the control of a lac promoter/operator/repressor system using transmission electron microscopy combined with energy dispersive X-ray analysis mapping.

Click Grafting of Alkyne-containing Vinyl Polymers onto Biosynthesized Extracellular Matrix Protein Containing Azide Functionality and Adhesion Control of Human Umbilical Vein Endothelial Cells.

In vivo incorporation of a phenylalanine (Phe) analogue, p-azidophenylalanine (p-N3Phe) into an artificial extracellular matrix protein (aECM-CS5-ELF) was accomplished using a bacterial expression host that harbors the mutant phenylalanyl-tRNA synthetase (PheRS) with an enlarged binding pocket, in which the Ala294Gly/Thr251Gly mutant PheRS (PheRS**) was expressed under the control of T7 promoters. In this study, biosynthesized aECM-CS5-ELF containing p-N3Phe (aECM-CS5-ELF-N3) was coupled with alkyne-containing vinyl polymers prepared via controlled radical polymerization of three vinyl monomers, (styrene, acrylamide, and N-isopropylacrylamide) using a trithiocarbonate as the RAFT agent. Grafting of the vinyl polymers onto the aECM was accomplished via a copper-catalyzed alkyne-azide click reaction. The lower critical transition temperature (LCST) was evaluated, as well as the solubility in aqueous and organic media, which are dependent on the incorporation ratio of p-N3Phe and species of graft chains, in which the LCST behavior was altered remarkably when poly(N-isopropylacrylamide) moieties were attached as side chains. Circular dichroism measurements indicate conformational change was not induced by the grafting. Specific adhesion of human umbilical vein endothelial cells (HUVECs) onto the aECM-CS5-ELF-N3-graft-poly(N-isopropylacrylamide) composite surface and subsequent temperature-sensitive detachment were also demonstrated.

Design of electrophoretic and biocompatible poly(2-oxazoline)s initiated by perfluoroalkanesulfoneimides and electrophoretic deposition with bioactive glass.

N-Methyl bis[(nonafluorobutane)sulfonyl]imide (Nf2NMe) was synthesized to serve as an initiator for polymerization of 2-oxazolines and the polymerization activity and control of molecular weight were compared with conventional methyl triflate (TfOMe). Ring-opening polymerization of the vinyl-containing 2-oxazoline, 2-(3-butenyl)-2-oxazoline, and a subsequent thiol-ene click reaction and oxidation resulted in the successful synthesis of electrophoretic poly(2-oxazoline)s containing pendent sulfone. It was possible to coat the polymeric sulfones obtained via oxone oxidation (conversion, >99%) on a stainless-steel anode selectively. Furthermore, hybridization of the poly(2-oxazoline)s with bioactive glass (Bioglass45S5) by electrophoretic deposition (EPD) was investigated, and the biocompatibility of the hybrid was also evaluated.

Site-specific bioconjugation of a murine dihydrofolate reductase enzyme by copper(I)-catalyzed azide-alkyne cycloaddition with retained activity.

Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is an efficient reaction linking an azido and an alkynyl group in the presence of copper catalyst. Incorporation of a non-natural amino acid (NAA) containing either an azido or an alkynyl group into a protein allows site-specific bioconjugation in mild conditions via CuAAC. Despite its great potential, bioconjugation of an enzyme has been hampered by several issues including low yield, poor solubility of a ligand, and protein structural/functional perturbation by CuAAC components. In the present study, we incorporated an alkyne-bearing NAA into an enzyme, murine dihydrofolate reductase (mDHFR), in high cell density cultivation of Escherichia coli, and performed CuAAC conjugation with fluorescent azide dyes to evaluate enzyme compatibility of various CuAAC conditions comprising combination of commercially available Cu(I)-chelating ligands and reductants. The condensed culture improves the protein yield 19-fold based on the same amount of non-natural amino acid, and the enzyme incubation under the optimized reaction condition did not lead to any activity loss but allowed a fast and high-yield bioconjugation. Using the established conditions, a biotin-azide spacer was efficiently conjugated to mDHFR with retained activity leading to the site-specific immobilization of the biotin-conjugated mDHFR on a streptavidin-coated plate. These results demonstrate that the combination of reactive non-natural amino acid incorporation and the optimized CuAAC can be used to bioconjugate enzymes with retained enzymatic activity.

Site-specific fatty acid-conjugation to prolong protein half-life in vivo.

Therapeutic proteins are indispensable in treating numerous human diseases. However, therapeutic proteins often suffer short serum half-life. In order to extend the serum half-life, a natural albumin ligand (a fatty acid) has been conjugated to small therapeutic peptides resulting in a prolonged serum half-life via binding to patients' serum albumin in vivo. However, fatty acid-conjugation has limited applicability due to lack of site-specificity resulting in the heterogeneity of conjugated proteins and a significant loss in pharmaceutical activity. In order to address these issues, we exploited the site-specific fatty acid-conjugation to a permissive site of a protein, using copper-catalyzed alkyne-azide cycloaddition, by linking a fatty acid derivative to p-ethynylphenylalanine incorporated into a protein using an engineered pair of yeast tRNA/aminoacyl tRNA synthetase. As a proof-of-concept, we show that single palmitic acid conjugated to superfolder green fluorescent protein (sfGFP) in a site-specific manner enhanced a protein's albumin-binding in vitro about 20 times and the serum half-life in vivo 5 times when compared to those of the unmodified sfGFP. Furthermore, the fatty acid conjugation did not cause a significant reduction in the fluorescence of sfGFP. Therefore, these results clearly indicate that the site-specific fatty acid-conjugation is a very promising strategy to prolong protein serum half-life in vivo without compromising its folded structure and activity.

Synthesis of Cell-Penetrating S-Galactosyl-Oligoarginine Peptides as Inducers of Recombinant Protein Expression under the Control of lac Operator/Repressor Systems.

The synthesis of glycodendrimers and glycopoly(oxazoline)s as inducers of recombinant protein expression has recently been reported; however, these compounds induced the expression of only small amounts of the green fluorescence protein (GFP), which was used as the model recombinant protein, because of their poor ability to penetrate the Escherichia coli cell membrane. Therefore, S-galactosyl-oligo(Arg) conjugates have now been synthesized to overcome this problem. Following in vivo expression of GFP induced by each of the S-galactosyl (Arg)n constructs (n=5, 6, 8) at the T5 promoter in E. coli for 18 hours, we visually observed that the cultures fluoresced green light when excited with UV light. The fluorescent intensities for these cultures were greater than that found for a control culture, which indicates that the peptides had induced GFP expression. Quantitative fluorescent measurements also supported the observations that the peptides were better inducers of GFP expression than the galactosyl dendrimers and the poly(oxazoline)s and the natural inducer lactose. Because the level of GFP expression was directly related to the number of arginine moieties in each peptide, we propose that the number of arginine moieties is responsible for how well each peptide passes through the E. coli membrane, which affects the expression level. A similar tendency was observed when the T7 promoter was placed upstream from the gene for an artificial extracellular matrix protein and the S-Gal-oligo(Arg) peptides were used as inducers. To assess how the distance between two galactosyl moieties as well as how the multivalent effect (cluster effect) in an oligo(Arg) inducer affects the expression level of GFP, we synthesized a conjugate of Lys(Arg)8 (Lys=lysine) and two S-galactosyls, which enhanced the expression of GFP in comparison with that obtained for S-Gal(Arg)8 .

Dehydration polycondensation of dicarboxylic acids and diols using sublimating strong brønsted acids.

We investigated catalytic activities of strong brønsted acids for dehydration polycondensations of dicarboxylic acids and diols, which were carried out at low temperature (<100 °C) under reduced pressure (0.3-3 mmHg). Strong Brønsted acids, bis(perfluoroalkanesulfonyl)imide and perfluoroalkanesulfonic acid, showed higher activity than p-toluenesulfonic acid or rare-earth catalysts at 60 °C. In particular, bis(nonafluorobutanesulfonyl)imide (Nf(2)NH) showed the highest activity to synthesize not only aliphatic polyester (M(n) > 19000) but also aromatic polyester (M(n) > 7000). The used Nf(2)NH was sublimated from the reaction flask during polycondensation, and the sublimate, Nf(2)NH, was extra pure so that we can reuse the catalyst without loss of the activity in the dehydration polycondensations.

Artificial extracellular matrix proteins containing phenylalanine analogues biosynthesized in bacteria using T7 expression system and the PEGylation.

In vivo incorporation of phenylalanine (Phe) analogues into an artificial extracellular matrix protein (aECM-CS5-ELF) was accomplished using a bacterial expression host that harbors the mutant phenylalanyl-tRNA synthetase (PheRS) with an enlarged binding pocket. Although the Ala294Gly/Thr251Gly mutant PheRS (PheRS**) under the control of T5 promoter allows incorporation of some Phe analogues into a protein, the T5 system is not suitable for material science studies because the amount of materials produced is not sufficient due to the moderate strength of the T5 promoter. This limitation can be overcome by using a pair of T7 promoter and T7 RNA polymerase instead. In the T7 expression system, it is difficult, however, to achieve a high incorporation level of Phe analogues, due to competition of Phe analogues for incorporation with the residual Phe that is required for synthesis of active T7 RNA polymerase. In this study, we prepared the PheRS** under T7 promoter and optimized culture condition to improve both the incorporation level of recombinant aECM protein and the incorporation level of Phe analogues. Incorporation and expression levels tend to increase in the case of p-azidophenylalanine, p-iodophenylalanine, and p-acetylphenylalanine. We evaluated the lower critical transition temperature, which is dependent on the incorporation ratio and the turbidity decreased when the incorporation level increased. Circular dichromism measurement indicated that this tendency is based on conformational change from random coil to ß-turn structure. We demonstrated that polyethylene glycol (PEG) can be conjugated at reaction site of Phe analogues incorporated. We also demonstrated that the increased hydrophilicity of elastin-like sequences in the aECM-CS5-ELF made by PEG conjugation could suppress nonspecific adhesion of human umbilical vein endothelial cells (HUVEC).

Click polyester: synthesis of polyesters containing triazole units in the main chain by click chemistry and improved thermal property.

A "click" polymerization of dialkynes that contain an ester linkages and diazides to has been performed to synthesize various polyesters, termed "click polyesters" with a high $\overline M _{\rm n}$ of 1.0 × 10(4) to 7.0 × 10(4) in an excellent yield. This polymerization accompanied a formation of 1,4-disubstituted triazoles in the polyester main chain by a Cu(I) catalyst. The triazole ring formation in the polyester main chain leads to improved thermal properties and enhancement of the even-odd effect of methylene chain length of the produced click polyesters. This report is the first report of the application of click chemistry to synthesize a series of polyesters under mild conditions.

Preparation and biodegradation of sugar-containing poly(vinyl acetate) emulsions.

To accelerate the biodegradability of poly(vinyl acetate)-based emulsions, emulsion copolymerizations of vinyl sugars, including triacetylated N-acetyl-D-glucosamine (GlcNAc)-substituted 2-hydroxyethyl methacrylate (GlcNAc(Ac)3-substituted HEMA), glucose-substituted HEMA (GEMA) and 6-O-vinyladipoyl-D-glucose (6-O-VAG) with vinyl acetate (VAc), were carried out using poly(vinyl alcohol) as an emulsifying agent in the presence of poly[(butylene succinate)-co-(butylene adipate)] [poly(BS-co-BA)]. Copolymerization with GEMA produced a stable emulsion and that with 6-O-VAG also produced a homogeneous emulsion. Their biodegradation tests indicated that PVAc main chain scission was accelerated by copolymerization with vinyl sugars.

Synthesis of sugar-substituted poly(phenylenevinylene)s.

Sugar-containing PPVs, poly{2-[O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)]-5-methoxy-p-phenylenevinylene-alt-p-phenylenevinylene} (PPV-GlcNAc) and poly{2,5-bis-[O-(beta-d-glucopyranosyl)]-p-phenylenevinylene-alt-p-phenylenevinylene} (PPV-Glc(2)), were synthesized via Heck reaction of p-divinylbenzene (DVB) with O-glycosylated hydroquinones, 2,5-dibromo-4-methoxyphenyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-beta-D-glucopyranoside (3) and 1,4-bis(O-2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-2,5-dibromobenzene (6), respectively. Acetyl protecting groups of the PPVs are completely removable under mild conditions (yield 69-87%). The structures were confirmed using (1)H NMR and IR spectra. Size exclusion chromatography (SEC) (eluent: DMF, polystyrene standards) measurements indicated that respective M(n) and M(w)/M(n) values of the obtained polymers are 4.49 x 10(3) and 2.3(5) (PPV-GlcNAc) and 3.86 x 10(3) and 1.3(9) (PPV-Glc(2)). These sugar-containing PPVs are soluble in water/DMF (8/2, v/v) and are recognized by Concanavalin A (Con A), D-glucose-binding protein. Blue shift of lambda(max) of the conjugated polymer backbone was confirmed when the glucose-substituted PPV interacts with Con A. Based on those binding properties, these results revealed that the obtained PPVs with pendant sugars have capabilities for detection of biological stimuli.

Polycondensation of dicarboxylic acids and diols in water catalyzed by surfactant-combined catalysts and successive chain extension.

Direct dehydration polycondensation of dicarboxylic acids and alcohols was carried out by surfactant-combined Brønsted and Lewis acids. This procedure did not require the removal of water, because the esterification was established at the interface of the emulsion in water. Emulsion polycondensations of 1,9-nonanediol (1,9-ND) and dodecanedioic acid (DDA) (the molar ratio of dicarboxylic acid to diol = 1:1) were carried out at 80 degrees C for 48 h in the presence of 16 wt % DBSA. The corresponding polyester (M(w) = 10.1 x 10(3)) was obtained in an excellent yield (99%). Chain extension in the emulsion was carried out using hexamethylene diisocyanate as the chain extender. SEC measurements indicated the expected shift to higher molecular weight region (M(w) = 11.4 x 10(3), M(w)/M(n) = 3.4) compared with parent polyester (M(w) = 4.5 x 10(3), M(w)/M(n) = 2.2).

Three-dimensional arrangement of sugar residues along helical polypeptide backbone. 2. Synthesis of periodic N-glycosylated peptides by polymerization of tripeptide active esters containing alpha,alpha-disubstituted amino acid.

New type of N-glycosylated peptides having periodic sequence of -[X-Gln(beta-D-GlcNAc)-Aib]- [X = L-Glu(OMe), L-Lys(Ac), L-Ala; Aib = alpha-aminoisobutyric acid] were synthesized by polymerization of glycosylated tripeptides with an active ester methods using Cl(-+)H(3)N-L-Glu(OMe)-Gln[beta-D-GlcNAc(Ac)(3)]-Aib-ONp (Np=p-nitrophenyl) (13a), Cl(-+)H(3)N-L-Lys(Ac)-Gln[beta-D-GlcNAc(Ac)(3)]-Aib-ONp (13b), and Cl(-+)H(3)N-L-Ala-Gln[beta-D-GlcNAc(Ac)(3)]-Aib-ONp (13c) as the monomers. Polymerizable glycosylated tripeptides were prepared by stepwise N,N-dicyclohexylcarbodiimide (DCC)/1-hydroxybenzotriazole (HOBt) method. Polymerizations of 13a-c were initiated by triethylamine and proceeded in DMSO at 50 degrees C for 5 days in the presence of 1-hydroxy-7-azabenzotriazole (HOAt) as the activator (conversions were 25-75%). The glycopeptides were deacetylated by hydrazine monohydrate in methanol to afford periodic glycopeptides 14 (12-27 residues) without racemization (yield, 35-89%). CD spectra in methanol, trifluoroethanol, and water of deacetylated glycopolymers 14a, 14b, and 14c showed double minima (206 and 222 nm) of negative Cotton effect indicating that N-glycoside (N-acetyl-d-glucosamine) was arranged three-dimensionally along the alpha-helical peptides in water as well as in organic protic solvents. The helix content depends on the solvent, peptide sequence, and spacer between peptide backbone and sugar. Interaction of the glycopeptides with wheat germ agglutinin (WGA) lectin was investigated by fluorescence measurement.

Asymmetric epoxidation of alpha-olefins having neighboring sugar chiral templates and alternating copolymerization with dicarboxylic anhydrides.

Sugar-substituted epoxides 5-8 were synthesized by asymmetric epoxidation (in CH(2)Cl(2)/water) of alpha-olefins having neighboring sugars (1-4) by use of an achiral oxidant (MCPBA), in which the sugar moiety acted as a chiral template. The diastereoselectivities depend on the methylene spacer between vinyl group and carbohydrate derivatives. The methylene spacer between sugar and vinyl groups influenced the diastereoselectvity. In the case of epoxidation of 4 at 27 degrees C for 24 h, the diastereoselectivity was the highest (99/1). Copolymerizations of 5-8 with succinic anhydride were attained at 100 degrees C for 72 h to give poly(ethylene succinate) having pendant carbohydrate [poly(SAn-alt-5), M(n) = 1.4 x 10(3); poly(SAn-alt-6), M(n) = 2.2 x10(3); poly(SAn-alt-7), M(n) = 2.9 x 10(3); poly(SAn-alt-8), M(n) = 1.8 x 10(3)]. The methylene spacer between sugar and epoxide has an effect on the reactivity of epoxide in copolymerization as well as the diastereoselectivity. Alternating copolymerization of 7 and glutaric anhydride gave a polyester of M(n) 4.2 x10(3).

Accelerated biodegradation of poly(vinyl alcohol) by glycosidations of the hydroxyl groups or addition of sugars.

Biodegradabilities of N-acetyl-d-glucosamine (GlcNAc)- (1) and chitobiose-substituted (2) poly(vinyl alcohol)s (PVA)s in a soil suspension (pH 6.5) were investigated at 25 degrees C for 40 days. Biochemical oxygen demand of 1 with a degree of substitution of 0.2-0.3 (DP = 430-480) was higher than that of PVA under the degradation condition. Size exclusion chromatography, (1)H NMR, and Fourier-transform infrared measurements of the recovered sample indicated that biodegradation of the PVA main chain was accelerated by partial glycosidation of hydroxyl groups in PVA. Similar acceleration was observed in a PVA/GlcNAc (50:50, w/w) mixture. Microbes which relate with degradation of the glycosidated polymers were grown in a culture medium including the soil suspension and the polymer as the carbon source. Polyacrylamide gel electrophoresis (SDS-PAGE) and IR measurements indicated that a cell-free extract derived from GlcNAc-substituted PVA was different from that in the PVA/GlcNAc mixture. The results suggested that the PVA main chain in GlcNAc-substituted PVA was cleaved by a different microorganism or via a mechanism different from that in the mixture. Chitobiose-substituted PVA 2 showed more enhanced acceleration, indicating that the sugar length influenced the degradability.

Noncovalent chiral domino effect on one-handed helix of nonapeptide containing a midpoint L-residue.

We here clarify whether noncovalent chiral domino effect characterized by the terminal interaction of a helical peptide with a chiral small molecule can alter the helical stability of N-deprotected peptides containing an L-residue covalently incorporated into the inner position. Two nonapeptides consisting of the midpoint L-leucine (1) or L-phenylalanine (2) and the achiral helix-forming residues were employed. NMR and IR spectroscopy and energy calculation indicated that both peptides adopt a 3(10)-helical conformation in chloroform. They strongly preferred a right-handed screw sense because of the presence of the midpoint L-residue. These original right-handed screw senses were retained on addition of chiral Boc-amino acid, but their helical stabilities clearly depended on its added chirality. Here, Boc-L-amino acid stabilizes the original right-handed helix, whereas the corresponding Boc-D-amino acid tends to less stabilize or destabilize it. This tendency was not observed for the corresponding N-Boc-protected peptides 1 and 2, strongly suggesting that the N-terminal amino group is required for controlling the stabilization of the original right-handed helix. Therefore, noncovalent chiral domino effect in peptides 1 and 2 can contribute even to the helical stability of a chiral peptide prevailing one-handed helix strongly through the midpoint L-residue. In addition, the N-terminal moiety of a 3(10)-helical peptide was found to generate chiral discrimination in complexation process with racemic additives.

Equivalent cross-relaxation rate imaging in the synthetic copolymer gels and invasive ductal carcinomas of the breast.

The values of equivalent cross-relaxation rate (ECR) correlated well with [i] water conditions in various copolymer gels and [ii] nature of malignant cells with regard to nuclear dysplasia and mitotic potential in breast carcinomas. The synthetic copolymer gels composed of any two or three monomers among 2-hydroxyethyl methacrylate (HEMA), glycidyl methacrylate (GMA), N-vinyl-2-pyrrolidinone (N-VP), methyl methacrylate (MMA) and benzyl methacrylate (BMA). The ECR measurement was performed by using an off-resonance saturation pulse under conventional field-echo imaging at frequency within +/- 75 ppm apart from the water resonance frequency. The ECR values were readily to determine and non-time consuming parameter for cross relaxation rate. The ECR values at the frequency offset by 7-ppm (ECR-7) were divided the sample gels two classes, which must correspond to hydrophilic or hydrophobic ones. The sensitivity in the gels was nearly equivalent to the cross-relaxation rate itself. In the breast carcinomas, the ECR-7 correlates with the nature of malignant cells with regard to nuclear dysplasia and mitotic potential. The ECR-7 is better or more accurate than the STR-7 because the SDNRs between carcinoma and glandular tissue increased by approximately 50% on the ECR-7 compared with the STR-7. Thus the ECR values could be a new parameter for malignancy and cell proliferative activity of the breast carcinomas with non-invasive modalities by magnetic resonance imaging.