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.     

Supramolecular Ligands for Histone Tails by Employing a Multivalent Display of Trisulfonated Calix[4]arenes

Post‐translational modification of histone tails plays critical roles in gene regulation. Thus, molecules recognizing histone tails and controlling their epigenetic modification are desirable as biochemical tools to elucidate regulatory mechanisms. There are, however, only a few synthetic ligands that bind to histone tails with substantial affinity. We report CA2 and CA3, which exhibited sub‐micromolar affinity to histone tails (especially tails with a trimethylated lysine). Multivalent display of trisulfonated calix[4]arene was important for strong binding. CA2 was applicable not only to synthetic tail peptides but also to endogenous histone proteins, and was successfully used to pull‐down endogenous histones from nuclear extract. These findings indicate the utility of these supramolecular ligands as biochemical tools for studying chromatin regulator protein and as a targeting motif in ligand‐directed catalysis to control epigenetic modifications.     

Heterogeneous decomposition of CHF2OCH2CF3 and CHF2OCH2C2F5 over various standard aluminosilica clay minerals in air at 313 K

The heterogeneous decomposition of CHF2OCH2C2F5, a potential substitute for hydrofluorocarbons, over aluminosilica clay minerals in air, was confirmed to occur at 313 K in a closed-circulation reactor. HC(O)OCH2C2F5, the gaseous main product was produced through hydrolytic elimination of F atoms from the CHF2OCH2- group. CHF2OCH2CF3 also decomposed to HC(O)OCH2CF3 over the clay minerals. The pseudo-first-order rate constants were determined for the decompositions over eight types of clay minerals (19 samples). The various clay minerals had different abilities to decompose these hydrofluoroethers. The decomposition rates per Brunauer-Emmett-Teller surface area and the conversion ratios to HC(O)OCH2C2F5 or HC(O)OCH2CF3 for the reactions over kaolinite, halloysite, and illite were high in comparison to those for the same reactions over montmorillonite, hectorite, and nontronite. The dependence of this heterogeneous reaction on temperature and relative humidity indicates that, in the environment, the reaction could be important only in hot, dry regions. The results did not suggest that sunlight would directly accelerate the decay of CHF2OCH2CF3 or CHF2OCH2C2F5. In the presence of clay-containing soils in arid areas, this hydrolytic oxidation reaction may significantly affect both the lifetime and the degradation products of CHF2OCH2CF3 and CHF2OCH2C2F5 in the troposphere.     

Hydrodynamic characteristics of a butterfly valve — Prediction of pressure loss characteristics

Investigation on fluidmechanic performance on a butterfly valve has been carried out. A practical valve model of a given thickness and a hub is used for the loss coefficient theory. A theoretical loss coefficient has been formulated from a contraction factor obtained by applying the generalized Borda mouthpiece theory.

Cavitation stages (such as cavitation inception, supercavitation inception, cavitation damage inception, choking cavitation) have been theoretically predicted from the valve loss coefficient. The cavitation prediction has been carried out by applying the free streamline theory, where the relation between the loss coefficient and the critical cavitation factor has been formulated. The results of the theoretical prediction equations agree well with the experimental results.     

Characterization of alternative plasticizers in poly(vinyl chloride) sheets for blood containers

This study aimed to optimize the ratio of dioctyl 4‐cyclohexene‐1,2‐dicarboxylate (DOTH) and di‐isononyl‐cyclohexane‐1,2‐dicarboxylate (DINCH^®) for use as plasticizers in poly(vinyl chloride) (PVC) sheets. We also evaluated the biological safety of DOTH for its potential to be part of a safe PVC‐based blood container. The suppression of hemolysis in mannitol‐adenine‐phosphate / red cell concentrates (MAP/RCC) with DOTH/(DINCH^®‐PVC) sheets and the elution of plasticizers from the sheets increased with higher DOTH compositions. The properties of the PVC sheet containing DOTH and DINCH^® in the ratio of 25:33 parts against PVC 100 parts as a weight were almost identical to the PVC sheet made of di(2‐ethylhexyl) phthalate. From a subchronic toxicity test, DOTH did not show any adverse effects on all organs, including the testes, epididymis, liver, and kidneys. The no‐observed‐adverse‐effect level was 300 mg/kg body weight/day in a rat. These results suggest that DOTH/DINCH^® (25:33) is a promising candidate for the replacement of di(2‐ethylhexyl) phthalate in blood containers. J. VINYL ADDIT. TECHNOL., 22:520–528, 2016. © 2015 Society of Plastics Engineers     

Amino acid sequences of hemoglobin beta chains of five species of pinnipeds: Neophoca cinerea, Otaria byronia, Eumetopias jubatus, Pusa hispida, and Pagophilus groenlandica

Pinnipeds (Otariidae, Odobenidae, and Phocidae) in the order Carnivora have one or two types (Hb I and Hb II) of hemoglobin components. These hemoglobins consist of identical beta chains and different alpha chains. We determined the complete amino acid sequences of the hemoglobin beta chain of three species of Otariidae (Australian sea lion, South American sea lion, and northern sea lion) and two species of Phocidae (ringed seal and harp seal) from intact beta chain and chemical cleavage fragments. The sequences are similar to beta chains of the already known sequences of pinnipeds. These sequences were compared with those of other carnivores (Mustelidae, Ursidae, Canidae, and Felidae) and adult human hemoglobin beta chain. Using Artiodactyla (pig) as an outgroup, we find that the tree constructed by means of phylogenetic analysis shows that Odobenidae is closest to Otariidae, and that Otariidae and Odobenidae are closer to Mustelidae than to Phocidae.     

Cadmium Purification with a Vibrating Reactor

This paper describes the results of pilot plant testwork on a novel single stage zinc solution purification process using a vibratory reactor. This technique was modified for continuous cadmium cementation. There are distinct advantages to combining an intensely agitated reactor with a high mass of precipitant per unit volume in the form of granular zinc instead of zinc dust, including significant reduction of cementation agent consumption and substantial upgrading of the cadmium cement. This technique has the potential to minimize equipment, reduce plant size, and diminish operating costs.     

A versatile method for analysis of serine/threonine posttranslational modifications by β-elimination in the presence of pyrazolone analogues

Post-translational modifications (PTMs) of serine and threonine occur by diverse mechanisms, including phosphorylation, sulfation, and various types of sugar chain modifications, making characterization of the resulting structures very labor-intensive. Moreover, to fully understand the biological functions of PTMs, both the sites of modification and the modified structures must be analyzed. The present work describes a novel, versatile strategy in which the released O-glycan and the formerly glycosylated/phosphorylated peptide are labeled and thus amenable to further study. In this approach, glycopeptides/phosphopeptides are subjected to β-elimination in the presence of pyrazolone derivatives (BEP), which in the same reaction labels the formerly glycosylated/phosphorylated peptide. The reaction is essentially a β-elimination/Michael addition in which a carbon-carbon bond-forming Michael donor rather than a heteroatomic Michael donor is used. The O-glycans released upon BEP are recovered as bis-pyrazolone derivatives, without any detectable side reaction (peeling). Using this technique, the O-glycan profiles of model mucin-type glycoproteins were successfully analyzed. The BEP strategy discriminates between phosphorylated and GlcNAcylated peptides, since cleaved GlcNAc is detectable. In addition, both the released O-glycan and the formerly glycosylated peptide can be selectively labeled by different reagents via a β-elimination reaction performed in the presence of pyrazolone and the thiol Michael donor.     

Simultaneous analysis of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan disaccharides by glycoblotting-assisted sample preparation followed by single-step zwitter-ionic-hydrophilic interaction chromatography

Glycosaminoglycans (GAGs) play important roles in cell adhesion and growth, maintenance of extracellular matrix (ECM) integrity, and signal transduction. To fully understand the biological functions of GAGs, there is a growing need for sensitive, rapid, and quantitative analysis of GAGs. The present work describes a novel analytical technique that enables high throughput cellular/tissue glycosaminoglycomics for all three families of uronic acid-containing GAGs, hyaluronan (HA), chondroitin sulfate (CS)/dermatan sulfate (DS), and heparan sulfate (HS). A one-pot purification and labeling procedure for GAG Δ-disaccharides was established by chemo-selective ligation of disaccharides onto high density hydrazide beads (glycoblotting) and subsequent labeling by fluorescence. The 17 most common disaccharides (eight comprising HS, eight CS/DS, and one comprising HA) could be separated with a single chromatography for the first time by employing a zwitter-ionic type of hydrophilic-interaction chromatography column. These novel analytical techniques were able to precisely characterize the glycosaminoglycome in various cell types including embryonal carcinoma cells and ocular epithelial tissues (cornea, conjunctiva, and limbus).