Autologous Induced Stem-Cell-Derived Retinal Cells for Macular Degeneration.

We assessed the feasibility of transplanting a sheet of retinal pigment epithelial (RPE) cells differentiated from induced pluripotent stem cells (iPSCs) in a patient with neovascular age-related macular degeneration. The iPSCs were generated from skin fibroblasts obtained from two patients with advanced neovascular age-related macular degeneration and were differentiated into RPE cells. The RPE cells and the iPSCs from which they were derived were subject to extensive testing. A surgery that included the removal of the neovascular membrane and transplantation of the autologous iPSC-derived RPE cell sheet under the retina was performed in one of the patients. At 1 year after surgery, the transplanted sheet remained intact, best corrected visual acuity had not improved or worsened, and cystoid macular edema was present. (Funded by Highway Program for Realization of Regenerative Medicine and others; University Hospital Medical Information Network Clinical Trials Registry [UMIN-CTR] number, UMIN000011929 .).

Isolation and characterization of a novel peroxisomal choline monooxygenase in barley.

Glycine betaine (GB) is a compatible solute accumulated by many plants under various abiotic stresses. GB is synthesized in two steps, choline â†’ betaine aldehyde â†’ GB, where a functional choline-oxidizing enzyme has only been reported in Amaranthaceae (a chloroplastic ferredoxin-dependent choline monooxygenase) thus far. Here, we have cloned a cDNA encoding a choline monooxygenase (CMO) from barley (Hordeum vulgare) plants, HvCMO. In barley plants under non-stress condition, GB had accumulated in all the determined organs (leaves, internodes, awn and floret proper), mostly in the leaves. The expression of HvCMO protein was abundant in the leaves, whereas the expression of betaine aldehyde dehydrogenase (BADH) protein was abundant in the awn, floret proper and the youngest internode than in the leaves. The accumulation of HvCMO mRNA was increased by high osmotic and low-temperature environments. Also, the expression of HvCMO protein was increased by the presence of high NaCl. Immunofluorescent labeling of HvCMO protein and subcellular fractionation analysis showed that HvCMO protein was localized to peroxisomes. [(14)C]choline was oxidized to betaine aldehyde and GB in spinach (Spinacia oleracea) chloroplasts but not in barley, which indicates that the subcellular localization of choline-oxidizing enzyme is different between two plant species. We investigated the choline-oxidizing reaction using recombinant HvCMO protein expressed in yeast (Saccharomyces cerevisiae). The crude extract of HvCMO-expressing yeast coupled with recombinant BBD2 protein converted [(14)C]choline to GB when NADPH was added as a cofactor. These results suggest that choline oxidation in GB synthesis is mediated by a peroxisomal NADPH-dependent choline monooxygenase in barley plants.

Evaluation of Antioxidant Activity and Amino Acids in the Mucus of Mackerel for Cosmetic Applications.

To identify antioxidants for improving rough skin, we aimed to analyze the amino acid composition of fish mucus and antioxidant activity of the mucus component. Specifically, we aimed to examine the antioxidant properties of dialyzed mucus components secreted from mackerel, which can be used as raw materials for producing cosmetics. The amino acid composition of hydrolyzed mucus was examined by ultra-high-performance liquid chromatography after dialyzing mucus. In addition, the antioxidant activity of the mucus was evaluated via oxygen radical absorption capacity and Trolox equivalent antioxidant capacity assays. The amino acid composition differed between the low-molecular-weight and high-molecular-weight fractions. Moreover, the low-molecular-weight fraction of farmed mackerel mucus exhibited antioxidant activity with high specificity. The results suggest that antioxidant peptides or free amino acids are present in the low-molecular-weight fraction of farmed mackerel mucus.

Unusual behavior of CMC for binary mixtures of alkyltrimethylammonium bromides: dependence on chain length difference.

The critical micelle concentrations (CMC) of binary mixtures of alkyltrimethylammonium bromides (CnTAB) were measured by a conductivity method. The CMCs of C12TAB-C14TAB and C14TAB-C16TAB systems exhibit the usual behavior, namely a monotonic decrease of the CMC with the mole fraction of the longer chain surfactant. However, the CMC behaviors of C10TAB-C16TAB, C11TAB-C16TAB, C12TAB-C16TAB, and C11TAB-C14TAB are unusual. The behaviors of the CMCs with mole fraction for these systems consist of three regions, of which the first is characterized by a very small decrease of the CMC in the range of low mole fraction, followed by a second where there is an abrupt decrease of the CMC, and a third where the CMCs exhibit their usual behavior. The molecular interaction parameter omega is almost equal to zero for mixtures that have the usual CMC behavior, but is small and positive for those systems with unusual CMCs. We infer that for very low mole fractions of C16TAB, the C16TA ion in the C12TAB-C16TAB system penetrates imperfectly into the micelle and its two methylene groups exist outside the micelle.

Screening Evaluation of the Interaction of Linear-Chain or Branched-Chain Peptides with Multilamellar Vesicle, Using Confocal Laser Microscopy.

In the present research, we prepared an acidic liposome as a cell model and used confocal laser microscopy to evaluate its interaction with oligopeptides that had high membrane permeability or affinity. The results showed that, for short peptides of about ten residues, the positive charge peptides interacted with the acidic liposome strongly. For peptides that interacted with the liposome, no difference was found between linear-chain and branched-chain peptides due to their structure.

Critical micelle concentrations and interaction parameters of aqueous binary surfactant:ionic surfactant mixtures.

A relationship between the critical micelle concentration (CMC) and the surfactant's composition in the bulk phase that supercedes Rubingh's method is derived for aqueous mixtures of ionic surfactants by considering the interaction between a micellar ionic aggregate and the diffusion layer around it. To test this approach we measured the CMCs of solutions of cationic surfactant mixtures and also of alkylammonium dodecanesulfonate mixtures. In the absence of controlled concentration of the counterion, the CMCs do not fit Clint's equation, but CMCs measured at a constant counterion concentration fit it approximately. The interaction parameter in the theory of regular solutions is obtained from the relationship between the micellar and bulk compositions. The values of the interaction parameter and the concentration exponent change with the hydrophobicity of the counterion in mixtures of alkylammonium dodecanesulfonates. The micellar composition of dodecylammonium chloride and dodecyltrimethylammonium chloride mixtures depends very little on the counterion concentration. The interaction energy between the ammonium and trimethylammonium groups of the cationic surfactants is about -0.05kT on average and depends on the concentration of the counterion.