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Silencing Y-box binding protein 1 (YB-1) can be an excellent target for cancer therapy and many lung cancer cells express the polysaccharide-recognition receptor Dectin-1. We designed a Dectin-1 targeting vehicle delivering YB-1-antisense DNA. First, we selected five optimal antisense DNA sequences to silence YB-1 from among 153 candidates. We chose the sequence closest to the start codon (AS014), and attached dA40 to the 3' end; dA40 promotes complex formation with a β-(1➝3)-d-glucan called schizophyllan (SPG). The resultant complexes were applied to 12 human-oriented lung cancer cell lines, and cell viability was examined. The cell lines exhibited decreased viability and showed strong affinity to bind SPG, suggesting the AS014/SPG complex entered the cells via the Dectin-1 mediated pathway.

The spatial distribution of bromine-labeled hydrophobic molecules—bromobenzene (BrBz) and 4-bromobenzylalcohol (BrBzOH)—incorporated in polymer micelles of poly(N,N-dimethylaminoethyl methacrylate)-block-poly(methyl methacrylate) (PDMAEMA-b-PMMA) was investigated using anomalous small-angle X-ray scattering (ASAXS) near the Br K-edge. ASAXS analyses revealed that BrBzOH was homogeneously dispersed in the hydrophobic PMMA core of the PDMAEMA-b-PMMA micelle, whereas BrBz was excluded from the vicinity of the core–shell interface of the PMMA core. The results indicate that the spatial distribution of hydrophobic molecules in polymer micelles strongly depend on the polarity of the encapsulated species.

The lectin Dectin-1 is a good target for β-glucan-mediated drug delivery. Although many murine studies of Dectin-1 have been performed, its human analog has not been studied well in terms of being a drug delivery target. We thus analyzed human Dectin-1 cDNA obtained from chronic myelogenous leukemia-derived cells, CML-1, and confirmed the findings of previous studies that there are many isoforms of human Dectin-1 due to exon skipping, although murine Dectin-1 has only two forms. When we transfected the Dectin-1 gene into a non-Dectin-1-expressing cell line and examined cellular uptake of the antisense DNA/β-glucan complex, we confirmed that expression of the target gene was effectively suppressed through β-glucan/Dectin-1-mediated uptake. The present results suggest that the β-glucan complex would be an effective tool to deliver antisense oligonucleotide (AS-ODN) to Dectin-1-expressing cells not only for mice but also for humans.

The assembly of supramolecular polymer bottlebrushes in aqueous systems is, in most cases, associated with a lateral aggregation of the supramolecular building blocks in addition to their axial stacking. Here, it is demonstrated that this limitation can be overcome by attaching three polymer chains to a central supramolecular unit that possesses a sufficiently high number of hydrogen bonding units to compensate for the increased steric strain. Therefore, a 1,3,5-benzenetrisurea-polyethylene oxide conjugate is modified with different peptide units located next to the urea groups which should facilitate self-assembly in water. For a single amino acid per arm, spherical micelles are obtained for all three tested amino acids (alanine, leucine, and phenylalanine) featuring different hydrophobicities. Only a slight increase in size and solution stability of spherical micelles is observed with increasing hydrophobicity of amino acid unit. In contrast, introducing two amino acid units per arm and thus increasing the number of hydrogen bonds per unimer molecule results in the formation of cylindrical structures, that is, supramolecular polymer bottlebrushes, despite a suppressed lateral aggregation. Consequently, it can be concluded that the number of hydrogen bonds has a more profound impact on the resulting solution morphology than the hydrophobicity of the amino acid unit.

Recent studies show that calix[4]arene-based micelles are monodisperse with defined Nagg values chosen from 4, 6, 8, 12, 20, and 32. Interestingly, all these numbers coincide with the face numbers of Platonic solids, so they are called "Platonic micelles." As long as a certain geometric condition is fulfilled, any amphiphilic molecule can form a Platonic micelle. The preferred Nagg values are explained in relation to the mathematical Tammes problem, namely, how to obtain the best coverage of a sphere's surface with multiple identical circles. The coverage ratio can be calculated and produces maxima at 4, 6, 12, 20, and 32, coinciding with the observed Nagg values. In this feature article, Platonic micelles as well as their morphological transition by controlling pH, salt, temperature, and electrostatic interactions are summarized.

The mixture ratios were used as a factor for aggregation number (Nagg) determination in binary mixtures of calix[4]arene surfactants. Platonic micelles having different Nagg were prepared by adjusting the ratio of oppositely charged compounds.

Molecular parameters and chain stiffness of polyethylene glycol/α-cyclodextrin inclusion complex: polyrotaxane were determined by use of synchrotron small angle X-ray scattering combined with light scattering and viscometry coupled with SEC.

We evaluated structural factors characterizing PEG-b-P(Asp-Bzl) micelles including core size, aggregation number (Nagg), and core surface PEG density by means of small-angle X-ray scattering (SAXS), field flow fractionation with multi-angle light scattering (FFF-MALS) analysis, and DLS. Furthermore, we evaluated the stability of PEG-b-P(Asp-Bzl) micelles by means of GPC. This paper reports the correlation between the evaluated micelles' structural factors and the micelles' behaviors including the micelles' in vivo pharmacokinetic behaviors. One micelle PEG(12)-b-P(Asp-Bzl) (PEG=12,000) exhibited a high core surface density (~0.99 chain/nm(2)). In these circumstances, PEG(12)-b-P(Asp-Bzl) micelles exhibited a highly stretched PEG brush form. However, the evaluated core surface PEG densities could not fully explain the micelles' in vivo pharmacokinetic behaviors. In contrast, GPC will become a strong tool for predicting PEG(12)-b-P(Asp-Bzl) micelles' in vivo behaviors, as well as the micelles' in vitro behaviors. The stability results correlated strongly with the area-under-the-curve (AUC) values of PEG-b-P(Asp-Bzl) micelles' in vivo pharmacokinetics. Finally, we evaluated PEG(12)-b-P(Asp-Bzl) micelles' most effective structural factor for determining the micelles' behaviors, and the micelles' outermost shell surface's PEG density (DOS, PEG) correlated with the micelles' behaviors. We revealed that the evaluated DOS, PEG is the most important factor for understanding PEG(12)-b-P(Asp-Bzl) micelles' behaviors.

In this article, we have demonstrated that π–π stacking interactions can be synchronized through the rapid hydration of π-conjugated molecules, generating a sort of metastable state. The self-assem...

The aggregation behavior of the asymmetric cationic surfactant, hexyldimethyloctylammonium bromide (C6C8DAB), was analyzed by dynamic light scattering (DLS) and transmittance measurements at various surfactant concentrations. DLS measurements showed that large aggregates appeared spontaneously as the solution was diluted toward the critical micelle concentration (CMC), accompanied by the solution turning turbid.

Antisense oligonucleotides (AS-ODNs) hybridize with specific mRNAs, resulting in interference with the splicing mechanism or the regulation of protein translation. We previously demonstrated that the β-glucan schizophyllan (SPG) can form a complex with AS-ODNs with attached dA40 (AS-ODNs/SPG), and this complex can be incorporated into cells, such as macrophages and dendritic cells, expressing the β-glucan receptor Dectin-1. We have achieved efficient gene silencing in animal models, but the uptake mechanism and intracellular distribution are unclear. In this study, we prepared the complex consisting of SPG and AS-ODNs (AS014) for Y-box binding protein-1 (YB-1). After treatment with endocytosis inhibitor Pitstop 2 and small interfering RNA targeting Dectin-1, we found that AS014/SPG complexes are incorporated into cells by Dectin-1-mediated endocytosis and inhibit cell growth in a Dectin-1 expression level-dependent manner. After treatment with AS014/SPG complexes, we separated the cell lysate into endosomal and cytoplasmic components by ultracentrifugation and directly determined the distribution of AS014 by reverse transcription PCR using AS014 ODNs as a template or a reverse transcription primer. In the cytoplasm, AS014 clearly hybridized with YB-1 mRNAs. This is the first demonstration of the distinct distribution of the complex in cells. These results could facilitate the clinical application of the complex.

Correction for ‘A tetramer micelle: the smallest aggregation number corresponding to the vertex number of regular polyhedra in platonic micelles’ by Shota Fujii et al., Soft Matter, 2018, DOI: 10.1039/c7sm02028g.

The size, surface charge, and microstructure of lipoplexes comprising cationic lipids and nucleic acids are important factors for transfection efficiency. As these properties are largely determined by the cationic lipids used, a number of studies on the relationship between cationic lipids and the transfection efficiency have been reported. Among the many cationic lipids, lipids with multivalent cationic head groups are expected to be potent transfection reagents. Here, we prepared calix[4]arene-based lipids with different alkyl chain lengths from C3 to C15 and evaluated the relationship between the alkyl chain length and the transfection efficiency. C6 lipoplexes exhibited the highest transfection efficiency among all lipoplexes. The gene expression with C9 and C12 lipoplexes was slightly lower than that with C6 lipoplexes. C3 lipoplexes hardly induced gene expression, while C15 lipoplexes exhibited no complexation with plasmid DNA. Although all lipoplexes exhibited nearly identical characteristics, they exhibited different behaviours in terms of the interactions between the lipoplexes and anionic micelles comprising phosphatidylserine, a model of endosomal vehicle. After mixing with phosphatidylserine micelles, C6 lipoplexes released the bound plasmid DNA at pH 5 but not at pH 7, indicating that they can interact with the late endosomal membrane after being incorporated into cells. No plasmid DNA was released from C9 or C12 lipoplexes at either pH values. Thus, the alkyl chain length of cationic lipids is related to their interaction with the endosomal compartment and can provide a basis for the design of novel transfection reagents.

We investigated the cross-sectional structure of a molecular monolayer nanotube self-assembled from asymmetric bolaamphiphiles having two different hydrophilic headgroups. Small-angle X-ray scattering measurements clarified that the glucose and amino headgroups form the exterior and interior surfaces of the nanotube, respectively.

The aggregation number of classical micelles exhibits a certain distribution, which is a recognizable feature of conventional micelles. However, we recently identified perfectly monodisperse calix[4]arene-based micelles whose aggregation numbers agree with the vertex numbers of regular polyhedra, that is, Platonic solids, and thus they are named “Platonic micelles”. Regarding our hypothesis of the formation mechanism of Platonic micelles, both repulsive interactions including steric hindrance and electrostatic repulsions among the headgroups are important for determining their aggregation number; however, neither of these is necessarily needed to consider. In this study, we employed polyethylene glycols (PEGs) as the nonionic headgroup of calix[4]arene-based amphiphiles to study the effects of only repulsive interactions caused by steric hindrance on the formation of Platonic micelles. The amphiphiles containing relatively low-molecular-weight PEGs (550 or 1000 g mol–1) form dodecamer or octamer micelles,...

The efficient induction of antigen-specific immune responses requires not only promotion of the uptake of antigens and adjuvant molecules into antigen-presenting cells but also control of their intracellular behavior. We previously demonstrated that the β-glucan schizophyllan (SPG) can form complexes with CpG oligonucleotides with attached dA40 (CpG-dA/SPG), which can accumulate in macrophages in the draining inguinal lymph nodes and induce strong immune responses. In this study, we prepared various conjugates composed of antigenic peptide (OVA257–264) and dA40 and made complexes with SPG. The conjugates with a disulfide bond between OVA257–264 and dA40 were easily cleaved by glutathione. The resultant peptides with a hydrophobic amino acid at the C-terminal end was recognized by puromycin-insensitive leucine aminopeptidase (PILS-AP), which trims antigenic peptide precursors and prepares peptides of eight or nine amino acids in length, which is the optimal length for binding to major histocompatibility co...

Supramolecular assemblies have in the past been considered mechanically weak and in most cases unable to withstand their own weight. Calixarene-derived networks can, however, provide robust supramolecular gels. Incorporating a photoreactive stilbene moiety, we show that the aggregation state of the material can be tuned by heating and UV exposure in order to control the mechanical as well as the fluorescent properties. Regulating the extent of heating to control the proportion of H-aggregates and J-aggregates and further cross-linking of H-aggregates by control over UV exposure allows for adjustable photopatterning of the fluorescence as well as the material stiffness in the range from ∼100 to 450 kPa. We expect this straightforward supramolecular system will be suitable for advanced prototyping in applications where modulus and shape are important design criteria.

The concept of micelles was first proposed in 1913 by McBain and has rationalized numerous experimental results of the self-aggregation of surfactants. It is generally agreed that the aggregation number (Nagg) for spherical micelles has no exact value and a certain distribution. However, our studies of calix[4]arene surfactants showed that they were monodisperse with a defined Nagg whose values are chosen from 6, 8, 12, 20, and 32. Interestingly, some of these numbers coincide with the face numbers of Platonic solids, thus we named them “Platonic micelles”. The preferred Nagg values were explained in relation to the mathematical Tammes problem: how to obtain the best coverage of a sphere surface with multiple identical circles. The coverage ratio D(N) can be calculated and produces maxima at N = 6, 12, 20, and 32, coinciding with the observed Nagg values. We presume that this “Platonic nature” may hold for any spherical micelles when Nagg is sufficiently small.

Several immune system cell surface receptors are reported to be associated with osteoclastogenesis. Dectin 1, a lectin receptor for β-glucan, is found predominantly on cells of the myeloid lineage. In this study, we examined the effect of the dectin 1 agonist curdlan on osteoclastogenesis. In mouse bone marrow cells and dectin 1-overexpressing RAW 264.7 cells (d-RAWs), curdlan suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation, bone resorption, and actin ring formation in a dose-dependent manner. This was achieved within non-growth inhibitory concentrations at the early stage. Conversely, curdlan had no effect on macrophage colony-stimulating factor-induced differentiation. Furthermore, curdlan inhibited RANKL-induced nuclear factor of activated T cell cytoplasmic 1 (NFATc1) expression, thereby decreasing osteoclastogenesis-related marker gene expression, including tartrate-resistant acid phosphatase, osteoclast stimulatory transmembrane protein, cathepsin K, and matrix metallopeptidase 9. Curdlan inhibited RANKL-induced c-fos expression, followed by suppression of NFATc1 autoamplification, without significantly affecting the NF-κB signaling pathway. We also observed that curdlan treatment decreased Syk protein in d-RAWs. Inhibition of the dectin 1-Syk kinase pathway by Syk-specific siRNA or chemical inhibitors suppressed osteoclast formation and NFATc1 expression stimulated by RANKL. In conclusion, our results demonstrate that curdlan potentially inhibits osteoclast differentiation, especially NFATc1 expression, and that Syk kinase plays a crucial role in the transcriptional pathways. This suggests that the activation of dectin 1-Syk kinase interaction critically regulates the genes required for osteoclastogenesis.

Synchrotron small-angle X-ray scattering (SAXS) was measured from aqueous solutions of polyion complexes (PICs), which were prepared by mixing oppositely charged diblock copolymers MAPTAC-b-PMPC and AMPS-b-PMPC. MAPTAC represents polycationic (3-(methacryloylamino)propyl)trimethylammonium chloride, AMPS represents polyanionic sodium 2-(acrylamido)-2-methylpropanesulfonate and PMPC represents hydrophilic poly-zwitterion (2-(methacryloyloxy)ethyl phosphorylcholine). The degrees of polymerization of MAPTAC and AMPS were selected to be the same for each complex, and the ratio (Rp) of the polymerization degree for the cationic to neutral chains was changed from 0.27 to 9.5. The aggregation number (Nagg) of PIC was determined with light scattering combined with field-flow fractionation (FFF). The inner structures of PIC were determined by analyzing SAXS with the core-corona model. When the MPC chain was much longer than the charged ones (Rp=0.27), a star-like spherical conformation was formed. With an increase of Rp, Nagg increased, while the spherical form was maintained up to Rp=1.0. When Rp=4.7 and 9.5, the PIC became a worm-like cylinder and vesicle, respectively. Based on the determined structural parameters and aggregation numbers, the index to describe how the tethered PMPC chains were crowded on the shell was determined. It was observed that the crowding parameter was much smaller than that of poly(ethylene glycol in spherical core-shell polymeric micelles. Small-angle X-ray scattering (SAXS) was measured from the aqueous solutions of polyion complexes (PICs), prepared with isoelectrical mixing oppositely charged two block copolymers MAPTAC-b-PMPC and AMPS-b-PMPC. The ratio (Rp) of the polymerization degree of the cationic to the neutral chains was changed from 0.27 to 9.5. When Rp is 0.27, 4.7 and 9.5, PICs took a star-like spherical, a cylindrical and vesicle-like shape, respectively. We found how tethered PMPC chains are crowded on the shell, and this parameter was much smaller than those of PEG in spherical core-shell polymeric micelles.

Soft materials, such as polymers and fibers, are key industries in Japan and play a significant role in our daily lives [1–3]. These materials also contribute to our modern society in a wide range of applications, such as in aircrafts and automobiles, and in a diverse variety of medical, electronic, and optical devices. In the future, it is expected that polymer science and technology will continue to expand and play an important role in research related to the environment [4], energy science [5], green technology [6], and biotechnology [7].

A water soluble β-1,3-glucan schizophyllan (SPG) can be recognized by an immunocyte receptor called dectin-1. When we introduced naphthalene into the side chain of SPG (nSPG), it formed nanogel by physical cross-link and gained capability to ingest hydrophobic compounds such as doxorubicin. Our in vitro assay revealed that this nanogel can be used as specific delivery of anti-cancer drugs to immunocytes.

Accomplishing efficient delivery of a nanomedicine to the tumor site will encounter two contradictions as follows: 1) a contradiction between prolonged circulation time and endocytosis by cancer cells; 2) a dilemma between the stability of nanomedicine during blood circulation and intracellular drug release. While developing a nanomedicine which can solve the above two contradictions simultaneously is still a challenge, here, a multi-stimuli-responsive polymeric prodrug (PLys-co-(PLys-DA)-co-(PLys-SS-PTX))-b-PLGLAG-mPEG (P-PEP-SS-PTX-DA) is synthesized which is multi-sensitive to overexpressed matrix metalloproteinase-2 (MMP-2), low pH, and high concentration of glutathione in tumors. The P-PEP-SS-PTX-DA can be dePEGylated and reversed from negative at normal physiological pH to positive charge at tumor extracellular microenvironment; in this way, it can solve the contradiction between prolonged circulation time and endocytosis by cancer cells. Owing to the high reductive conditions in cancer cells, P-PEP-SS-PTX-DA is ruptured to release paclitaxel (PTX) intracellular efficiently; therefore, it can resolve the dilemma between the stability of nanomedicine during blood circulation and intracellular drug release. These indicate that the multi-stimuli-responsive polymeric prodrug has potential application prospects in drug delivery and cancer therapy.

Supramolecular polymerization of TPPS, a kind of self-assembling porphyrin, was investigated in a pressure-adjustable microflow channel. We found that the nucleation process of TPPS was accelerated...

Fine hydrogen-bonded networks over the micrometer scale were spontaneously constructed in microflow through the synchronized protonation of porphyrin having four carboxylic acid groups. The obtaine...

To date, little attention has been paid to the positive control of the self-assemble process itself. Rapid activation and synchronous growth of PCBM, a kind of fullerene derivative, in a precisely ...

CpG DNA, a ligand for Toll-like receptor 9 (TLR9), has been one of the most promising immunotherapeutic agents. Although there are several types of potent humanized CpG oligodeoxynucleotide (ODN), developing "all-in-one" CpG ODNs activating both B cells and plasmacytoid dendritic cells forming a stable nanoparticle without aggregation has not been successful. In this study, we generated a novel nanoparticulate K CpG ODN (K3) wrapped by the nonagonistic Dectin-1 ligand schizophyllan (SPG), K3-SPG. In sharp contrast to K3 alone, K3-SPG stimulates human peripheral blood mononuclear cells to produce a large amount of both type I and type II IFN, targeting the same endosome where IFN-inducing D CpG ODN resides without losing its K-type activity. K3-SPG thus became a potent adjuvant for induction of both humoral and cellular immune responses, particularly CTL induction, to coadministered protein antigens without conjugation. Such potent adjuvant activity of K3-SPG is attributed to its nature of being a nanoparticle rather than targeting Dectin-1 by SPG, accumulating and activating antigen-bearing macrophages and dendritic cells in the draining lymph node. K3-SPG acting as an influenza vaccine adjuvant was demonstrated in vivo in both murine and nonhuman primate models. Taken together, K3-SPG may be useful for immunotherapeutic applications that require type I and type II IFN as well as CTL induction.

We synthesized new calix[4]arene-based lipids, denoted by CPCaLn, bearing the choline phosphate (CP) group which is an inverse phosphoryl choline (PC) structure. Small-angle X-ray scattering and multi-angle light scattering coupled with field flow fractionation showed that these lipids form monodisperse micelles with a fixed aggregation number and diameters of 1.9 and 2.6 nm for lipids bearing C3 and C6 alkyl tails, respectively. Furthermore, when CPCaLn was mixed with the fluorescein isothiocyanate (FITC)-bearing lipids and added to cells, strong fluorescence was observed at 37 °C, but not at 4 °C, indicating that the micelles were taken up by the cells through endocytosis. Recent studies have shown that replacement of polymer-attached PC groups with CP groups markedly promotes cellular uptake, even though the surface charge is neutral. On the basis of the idea, CPCaLn micelles interacted with cells in the same way, suggesting that the micelles bearing CP groups are expected to use as carriers in the drug delivery system.

Anomalous small-angle X-ray scattering with two marker elements was applied to the structural analysis of poly(4-vinylphenol rubidium salt)-block-poly(4-bromostyrene) (RbPVPh-b-PBrS) micelles, where Br and Rb were the markers for the hydrophobic core and the hydrated corona, respectively. By using two different markers for the hydrophobic core and the hydrated corona, the form factors of the core and corona were extracted separately from the scattering profile of the whole RbPVPh-b-PBrS micelles. The form factor of the hydrophobic core (the spatial distribution of Br) revealed that the core was regarded as a solid sphere with a smooth surface and a radius of 47 nm. Conversely, the form factor of the spatial distribution of Rb+indicated that the shell of the RbPVPh-b-PBrS micelles was 15 nm thick.

A pair of oppositely charged diblock copolymers, poly(2-(methacryloyloxy)ethyl phosphorylcholine)-block-poly((3-(methacryloylamino)propyl)trimethylammonium chloride) (PMPC-b-PMAPTAC) and poly(2-(methacryloyloxy)ethyl phosphorylcholine)-block-poly(sodium 2-(acrylamido)-2-methylpropanesulfonate) (PMPC-b-PAMPS), was prepared via reversible addition-fragmentation chain transfer radical polymerization using a PMPC-based macro chain transfer agent. The pendant phosphorylcholine group in the hydrophilic PMPC block has anionic phosphate and cationic quaternary amino groups, which are neutralized within the pendant group. Therefore, the mixing of aqueous solutions of PMPC-b-PMAPTAC and PMPC-b-PAMPS leads to the spontaneous formation of simple core-shell spherical polyion complex (PIC) micelles comprising of a segregated PIC core and PMPC shells. The PIC micelles were characterized using (1)H NMR spin-spin (T2) and spin-lattice relaxation times (T1), diffusion-ordered NMR spectroscopy, static light scattering, dynamic light scattering (DLS), and transmission electron microscopy techniques. The hydrodynamic size of the PIC micelle depended on the mixing ratio of PMPC-b-PMAPTAC and PMPC-b-PAMPS; the maximum size occurred at the mixing ratio yielding stoichiometric charge neutralization. The PIC micelles disintegrated to become unimers with the addition of salts.

Lipopolysaccharide (LPS), the endotoxin of Gram-negative bacteria, is a strong elicitor in the immune system by interacting with lipopolysaccharide-binding protein and CD14 with high specificity. T...

RNA interference therapy utilizes physiological gene silencing that is originally found as a defense function against foreign RNAs. To silence the target gene, short double stranded RNA has to be delivered to cytosol. However, lack of a suitable delivering carrier is the major obstacle to practical usage. In this study, we present a novel complex consisting of β-1,3-glucan and short interference RNA (siRNA) as a solution for the problem. We used a β-1,3-glucan schizophyllan (SPG) and a siRNA (dA-siTNFα) that is designed to suppress tumor necrosis factor alpha (TNF-α), where the sense strand of siRNA has (dA(40)) tail to induce complexation with SPG. The dA-siTNFα/SPG complex showed higher affinity to recombinant dectin-1 than SPG itself, where dectin-1 is a β-1,3-glucan receptor expressed on antigen presenting cells and can be a target for specific delivery. The complex suppressed lipopolysaccharide (LPS)-induced TNF-α secretion by peritoneal macrophages in vitro. When the complex was intravenously injected, the oligonucleotide accumulated in liver; especially distributed into Kupffer cells. The complex significantly decreased the serum TNF-α level for the mouse model of LPS-induced acute hepatitis. This new siRNA delivery system may overcome the problem for RNA interference therapy because of its non-toxicity and high target specificity.

Shear-induced crystallization of a long chain hyper-branched polypropylene (LCB-PP, denoted PP-3) was carried out at a relatively high temperature of 170 °C, close to its melting temperature of 158 °C. Small-angle X-ray scattering (SAXS) showed that the intensity ratio of the normal to the perpendicular to the shear (V/H) was always larger than 1 for PP-3, indicating that shish-like structures were predominately formed and further growth to kebab was suppressed in PP-3. The crystallization behavior of PP-3 can be explained by the nature of PP-3 that there were a large amount of crystallization nuclei due to their branching points, and the point-like precursors formed from these nuclei were arrayed linearly along the shear direction and transformed into thread-like precursors or premature shishs; however, owing to the large amount of branching, further progress in crystallization was suppressed.

Cancer vaccine has the ability to directly eradicate tumor cells by creating and activating cytotoxic T lymphocytes (CTLs). To achieve efficient CTL activity and to induce Th1 responses, it is essential to administer an appropriate adjuvant as well as an antigen. CpG-ODN is known as a ligand of Toll-like receptor 9 (TLR9) and strongly induces Th1 responses. In our previous study, we developed a CpG-ODN delivery system by use of the formation of complexes between ODN and a β-glucan SPG, denoted as CpG/SPG, and demonstrated that CpG/SPG induces high Th1 responses. In this study, we created a nanogel made from CpG/SPG complexes through DNA-DNA hybridization (cross-linked (CL)-CpG). Immunization with CL-CpG induced much stronger antigen-specific Th1 responses in combination with the antigenic protein ovalbumin (OVA) than that with CpG/SPG. Mice preimmunized with CL-CpG and OVA exhibited a long delay in tumor growth and an improved survival rate after tumor inoculation. These immune inductions can be attributed to the improvement of cellular uptake by the combination of increased size and the cluster effect of the β-glucan recognition site in the nanogel structure. In other words, the particle nature of CL-CpG, instead of the semiflexible rod conformation of CpG/SPG, enhanced the efficacy of a cancer vaccine. The present results indicate that CL-CpG can be used as a potent vaccine adjuvant for the treatment of cancers and infectious diseases.

We explored in detail the relationship between the structure in aqueous solution and immunostimulatory activity of polypod-shaped DNAs, called polypodnas. The polypodnas were constructed using 3-6 oligodeoxynucleotides (ODNs) to obtain tri-, tetra-, penta-, and hexapodna, each of which had 3, 4, 5, and 6 arms made of double-stranded DNA, respectively. A highly potent immunostimulatory CpG sequence was included into each of the polypodnas. Synchrotron X-ray scattering analysis showed that the double-stranded DNA arms of all of the polypodnas adopted a B-form DNA conformation. The analysis also suggested that some nucleotides in the central parts of pentapodna and hexapodna did not form base pairs, whereas those of tripodna and tetrapodna all formed base pairs. This difference would occur because of an increase in steric hindrance and electrical repulsion with increasing number of arms. The pentapodna and hexapodna induced a large amount of tumor necrosis factor α-release from macrophage-like cells compared with the tripodna and tetrapodna, suggesting that the partly loosened DNA in polypodna with many arms is advantageous for exposing the immunostimulatory sequences of the polypodna.

The new experimental system has been launched by coupling with the measurement techniques, which are grazing incidence small/wide-angle X-ray Scattering (GISWAXS) and grazing incidence X-ray diffraction (GIXD), as well as X-ray reflectivity (XR), at the BL03XU of SPring-8. Using this integrated system, we can achieve to measure the hierarchical structure of the same sample.

Anomalous small-angle X-ray scattering (ASAXS) near the absorption edge of bromine has been used to investigate the internal structure of polymer micelles comprising poly(4-bromostyrene)-block-poly(ethylene glycol)-block-poly(4-bromostyrene) (PBrS-b-PEG-b-PBrS) in aqueous solution. The ASAXS analysis revealed that the hydrophobic core composed of PBrS can be regarded as a homogeneous sphere with a radius of 10.2 nm and a decaying electron density at the surface. The aggregation number of micelle directly estimated on the basis of ASAXS analysis well agreed with that determined by field-flow fractionation coupled with multiangle light scattering, confirming the reliability of ASAXS analysis. The small-angle X-ray scattering (SAXS) profile from whole PBrS-b-PEG-b-PBrS micelle was further analyzed by using the result of ASAXS analysis. The micelles could be well described by a core–corona model in which the monomer concentration of PEG in the shell decreased as R–4.2/3, where R is the distance from the cente...

Gene therapy by introducing a foreign DNA into a cell with defective gene has been expected to treat various incurable diseases such as cancers, inherited diseases, and cardiovascular disease. Cati...

Robotic technology is widening its scope into applications where robots coexist with humans and other living beings in a common space, and where cooperative and other physical interactions between the robots and the living beings are anticipated and actually occur. Safety of living beings and of robots in such an environment calls for solutions in robot mechanisms that are different from the recently used ones in industrial robotics. In this paper we present an actuator named a Twist Drive, which uses two strings that by twisting on each other generate a pulling force. The mechanism can be used to replace gear reducers in robot joints with benefits that originate in its muscle-like operation and in its nonlinear characteristics. A mathematical model to describe the kinematic transmission characteristics of the mechanism is presented and compared to experimental data. A passive characteristic of kinematic stiffness is discussed and evaluated in the paper. Experimental results show the adequacy of the propo...