Photoactivable Elimination of Tumorigenic Human Induced Pluripotent Stem Cells by Using a Lectin–Doxorubicin Prodrug Conjugate

Human pluripotent stem cells (hPSCs) are attractive resources for regenerative medicine, but medical applications are hindered by their tumorigenic potential. Previously, a hPSC-specific lectin probe, rBC2LCN, was identified through comprehensive glycome analysis by using high-density lectin microarrays. Herein, a lectin–doxorubicin (DOX) prodrug conjugate, with controllable photolysis activation for the elimination of tumorigenic human induced pluripotent stem cells, has been developed. rBC2LCN was fused with a biotin-binding protein, tamavidin (BC2Tama), and the fusion protein was expressed in Escherichia coli and purified by means of affinity chromatography. BC2Tama was then conjugated with doxorubicin-photocleavable biotin (DOXPCB). The BC2Tama–DOXPCB conjugates were observed to bind to hPSCs followed by internalization. Upon exposure to ultraviolet light, DOX was released inside the cells, which allowed specific killing of the hPSCs. Thus, BC2Tama–DOXPCB should be useful for the targeted elimination of hPSCs contained in hPSC-derived cell therapy products. This is the first report of the generation of lectin–prodrug conjugates. BC2Tama should be applicable for the targeted delivery of various types of biotinylated compounds into hPSCs.     

Exploring the Minimal RNA Substrate of Flexizymes

Flexizymes are tRNA acylation ribozymes that have been successfully used to facilitate genetic code reprogramming. They are capable of charging acid substrates onto various tRNAs and tRNA analogues. However, their minimal RNA substrate has not been investigated. Here we have designed fluorescently labeled short RNAs corresponding to the four, three, and two bases (4bRNA, 3bRNA, 2bRNA) at the tRNA 3′-end and explored the minimal RNA substrate of flexizymes, dFx and eFx. 3bRNA was the observed minimal RNA substrate of the flexizymes, but the efficiency of acylation of this short RNA was two to three times lower than that of 4bRNA. The efficiency of acylation of 4bRNA was comparable with that of the microhelix, a 22-base RNA conventionally used as a tRNA analogue for analyzing acylation efficiency. We also compared the efficiencies of acylation of the microhelix and 4bRNA with various acid substrates. Thanks to the short length of 4bRNA, its acyl-4bRNA products exhibited larger mobility shifts in gel electrophoresis than those exhibited by acyl-microhelix products with every substrate tested. This indicated that 4bRNA was an ideal RNA substrate for analyzing the efficiency of acylation by flexizymes.     

Designing block copolymer architectures for targeted membrane performance

Using a combination of block copolymer self-assembly and non-solvent induced phase separation, isoporous ultrafiltration membranes were fabricated from four poly(isoprene-b-styrene-b-4-vinylpyridine) triblock terpolymers with similar block volume fractions but varying in total molar mass from 43 kg/mol to 115 kg/mol to systematically study the effect of polymer size on membrane structure. Small-angle X-ray scattering was used to probe terpolymer solution structure in the dope. All four triblocks displayed solution scattering patterns consistent with a body-centered cubic morphology. After membrane formation, structures were characterized using a combination of scanning electron microscopy and filtration performance tests. Membrane pore densities that ranged from 4.53 × 10¹⁴ to 1.48 × 10¹⁵ pores/m² were observed, which are the highest pore densities yet reported for membranes using self-assembly and non-solvent induced phase separation. Hydraulic permeabilities ranging from 24 to 850 L m⁻² h⁻¹ bar⁻¹ and pore diameters ranging from 7 to 36 nm were determined from permeation and rejection experiments. Both the hydraulic permeability and pore size increased with increasing molar mass of the parent terpolymer. The combination of polymer characterization and membrane transport tests described here demonstrates the ability to rationally design macromolecular structures to target specific performance characteristics in block copolymer derived ultrafiltration membranes.     

The Attenuated Secretion of Hyaluronan by UVA-Exposed Human Fibroblasts Is Associated with Up- and Downregulation of HYBID and HAS2 Expression via Activated and Inactivated Signaling of the p38/ATF2 and JAK2/STAT3 Cascades

Little is known about the effects on hyaluronan (HA) metabolism of UVA radiation. This study demonstrates that the secretion of HA by human dermal fibroblasts (HDFs) is downregulated by UVA, accompanied by the down- and upregulation of mRNA and protein levels of the HA-synthesizing enzyme (HAS2) and the HA-degrading protein, HYaluronan Binding protein Involved in HA Depolymerization(HYBID), respectively. Signaling analysis revealed that the exposure distinctly elicits activation of the p38/MSK1/CREB/c-Fos/AP-1 axis, the JNK/c-Jun axis, and the p38/ATF-2 axis, but downregulates the phosphorylation of NF-kB and JAK/STAT3. A signal inhibition study demonstrated that the inhibition of p38 significantly abrogates the UVA-accentuated mRNA level of HYBID. Furthermore, the inhibition of STAT3 significantly downregulates the level of HAS2 mRNA in non-UVA exposed HDFs. Analysis using siRNAs demonstrated that transfection of ATF-2 siRNA but not c-Fos siRNA abrogates the increased protein level of HYBID in UVA-exposed HDFs. An inhibitor of protein tyrosine phosphatase but not of protein serine/threonine phosphatase restored the diminished phosphorylation level of STAT3 at Tyr 705, accompanied by a significant abolishing effect on the decreased mRNA expression level of HAS2. Silencing with a protein tyrosine phosphatase PTP-Meg2 siRNA revealed that it abrogates the decreased phosphorylation of STAT3 at Tyr 705 in UVA-exposed HDFs. These findings suggest that the UVA-induced decrease in HA secretion by HDFs is attributable to the down- and upregulation of HAS2 and HYBID expression, respectively, changes that are mainly ascribed to the inactivated signaling of the STAT3 axis due to the activated tyrosine protein phosphatase PTP-Meg2 and the activated signaling of the p38/ATF2 axis, respectively.     

NMR-Guided Repositioning of Non-Steroidal Anti-Inflammatory Drugs into Tight Junction Modulators

Bioavailability is a major bottleneck in the clinical application of medium molecular weight therapeutics, including protein and peptide drugs. Paracellular transport of these molecules is hampered by intercellular tight junction (TJ) complexes. Therefore, safe chemical regulators for TJ loosening are desired. Here, we showed a potential application of select non-steroidal anti-inflammatory drugs (NSAIDs) as TJ modulators. Based on our previous observation that diclofenac and flufenamic acid directly bound various PDZ domains with a broad specificity, we applied solution nuclear magnetic resonance techniques to examine the interaction of other NSAIDs and the first PDZ domain (PDZ1) of zonula occludens (ZO)-1, ZO-1(PDZ1). Inhibition of ZO-1(PDZ1) is expected to provide loosening of the epithelial barrier function because the domain plays a crucial role in maintaining TJ integrity. Accordingly, diclofenac and indomethacin were found to decrease the subcellular localization of claudin (CLD)-2 but not occludin and ZO-1 at the apicolateral intercellular compartment of Madin–Darby canine kidney (MDCK) II cells. These NSAIDs exhibited 125–155% improved paracellular efflux of fluorescein isothiocyanate insulin for the Caco-2 cell monolayer. We propose that these NSAIDs can be repurposed as drug absorption enhancers for peptide drugs.     

Novel Route to Propylene Carbonate: Selective Synthesis from Propylene Glycol and Carbon Dioxide

High surface area zirconium phosphate in an amorphous phase exhibits high activities for water-related reactions such as hydrolysis of ethyl acetate and esterification of acetic acid with ethanol. The zirconium phosphate is insoluble during the reaction, is recoverable by simple filtration, and can be reused at least five times without any treatment.     

Crosslinked acrylic pressure sensitive adhesives. 3. Effect of adherend on film formation

The reaction of isocyanate in pressure sensitive adhesive (PSA) films adhered on to various adherends having different surface tensions was monitored by depth profiling using attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy.In the latter stages of crosslinking reaction, unreacted isocyanate and its derivatives exist more in the bulk of the PSA than in the interfaces between PSA and adherends which are Teflon sheet and PE film having relatively lower surface tensions. In the case of using stainless steel having relatively higher surface tension as adherend, opposite segregation was observed compared to Teflon and PE.From X-ray photoelectron spectroscopy (XPS), it was revealed that N atoms exist more in surface than in bulk when stainless steel is used as adherend. We conclude that polyisocyanates migrate in the PSA film in order to minimize the magnitude of interfacial free energy between the PSA and the adherend, which leads to the change of surface tension of PSA film.     

Wien2wannier: From linearized augmented plane waves to maximally localized Wannier functions

We present an implementation of an interface between the full-potential linearized augmented plane wave package Wien2k and the wannier90 code for the construction of maximally localized Wannier functions. The FORTRAN code and a documentation is made available and results are discussed for SrVO₃, Sr₂IrO₄ (including spin-orbit coupling), LaFeAsO, and FeSb₂.     

DC‐linked hybrid generation system with an energy storage device including photovoltaic generation and gas engine cogeneration for residential houses

For the past few years, hybrid generation systems including solar panel and gas cogeneration have been used for residential houses. Solar panels can generate electronic power at daytime but not at night. But the power consumption of residential houses usually peaks in the evening. The gas engine cogeneration system can generate electronic power without such a restriction, and it also can generate heat power to warm up a house or to produce hot water. In this paper we propose a solar panel and gas engine cogeneration hybrid system with an energy storage device, combined by a DC bus. If a blackout occurs, the system still can supply electronic power for special house loads. We propose a control scheme for the system related to the charging level of the energy storage device and the voltage of the utility grid, which can be applied to both grid‐connected and standalone operation. Finally, we report experiments designed to demonstrate system operation and calculations for loss estimation. © 2012 Wiley Periodicals, Inc. Electr Eng Jpn, 182(4): 29–46, 2013; Published online in Wiley Online Library ( wileyonlinelibrary.com ).DOI 10.1002/eej.22321     

New dissimilarity measure for recognizing noisy subsequence trees

Tree is a data structure used to express various objects such as semistructured data and genes. When objects are represented as trees, computing tree similarity is essential for pattern recognition and retrieval. This paper considers the noisy subsequence tree recognition problem whose purpose is to recognize the original tree, given its noisy subsequence tree. Previous research on this problem relied on constrained tree edit distance to measure the dissimilarity. However, the number of relabelings must be predetermined to compute it. This paper proposes a new dissimilarity measure for this problem. Our dissimilarity measure is obtained by counting the node edit operations included in the unit‐cost tree edit distance that contribute to the matching of node labels. The number of relabelings need not be specified to compute our dissimilarity measure. Moreover, our measure achieves more accurate recognition performance and faster execution speed than the constrained tree edit distance. Our measure is also useful to solve the tree inclusion problem which is the problem of deciding whether a tree includes another tree and shows the extent of approximate tree inclusion when a tree incompletely includes another tree. © 2011 Wiley Periodicals, Inc.