Engineering microenvironmental cues for guiding stem cell fate

Injury, aging, and congenital disabilities of the muscular and neural systems impose a significant burden on patients and their families. Due to the tissue’s limited regenerative capacity, effective treatment interventions for restoring progressive damage is still lacking. Cell replacement therapy is primarily limited by the restricted supply of viable donor cells and variable graft survival. For addressing these limitations, we propose new strategies to obtain a target cell of interest from an autologous cell source. Herein, we engineer cell fate decisions by 1) harnessing host microenvironment and the CRISPR/dCas9-mediated transcriptional activation system to promote myogenesis of human endothelial progenitor cells (EPCs); and 2) employing substrate-mediated biophysical cues with soluble factors (biochemical cues) to drive cell commitment to neuronal lineages. For the first strategy, we hypothesized that therapeutic cells could be obtained in situ by employing the CRISPR/dCas9 system to engineer cell fate in the host tissue. Using this system, we transactivated MYOD1, a master regulator for myogenesis, to directly reprogram primary EPCs to skeletal myoblasts (SkMs). EPCs were chosen as a cell source for their easy accessibility, high proliferation, and potential contribution to regenerate vasculature and musculature tissue. The early myogenic commitment of EPCs was confirmed in vitro by MYOD1 expression, which yielded a 230-fold higher induction than the original EPCs. These cells were then transplanted for assessing their therapeutic efficacy in myotoxin-induced muscle injury model in immunodeficient mice. A one-month post-injury study resulted in the integration of induced SkMs to the injured host tissue, promotion of neoangiogenesis, and reduction in fibrotic scar formation. These findings indicate that CRISPR/Cas9-mediated target gene activation can be achieved in situ to accelerate muscle regeneration after myotoxin-induced damage. For the second strategy, we utilized both soluble and insoluble factors to convert the cell fate of neural stem/progenitor and somatic cells to various neuronal lineages, including motor neurons (MNs) and dopaminergic (DA) neurons. For soluble factors, cells were exposed to various biochemical factors, inspired by the neuronal niche environs during the natural developmental process. For insoluble factors, the conductive graphene substrate was used to support the endogenous electrical signal between neurons for enhancing the neuronal phenotypes and their functionality. We postulated that exposing the cells to these collective stimuli in vitro can alter their intrinsic signaling pathway to tailor their fate to neuronal lineages. To test the hypothesis, neural stem/progenitor and somatic cells were cultured on various substrates with or without electroactive graphene and aligned patterns. After two weeks to one month of cell fate induction in the chemically defined conditions, our results implied that cell adhesion, survival, neurite outgrowth, and maturation were facilitated on the electroactive substrates with aligned patterns compared to the control platforms. Taken together, our results in this dissertation demonstrate the feasibility of tailoring the donor cell fates within or across the germ layers. We achieved this by employing a transcriptional gene activation system and tunable microenvironmental cues elicited by soluble (chemical and growth factors) and insoluble (physical cues from the substrate) factors. Utilizing such strategies hold great promise for elucidating the optimal conditions to guide cell fate to target lineages. This work provides a rational basis for establishing a robust protocol and an in vitro culture platform to module cell fate decisions that could help realize the autologous cell-based therapy for muscular and neurodegenerative diseases

ADIPOQ Gene Variants Associated with Susceptibility to Obesity and Low Serum Adiponectin Levels in Healthy Koreans

OBJECTIVES: This study aimed to measure the association between the adiponectin, C1Q and collagen domain-containing (ADIPOQ) gene variants and obesity in Koreans. METHODS: Three single nucleotide polymorphisms located in the ADIPOQ gene were genotyped in a population-based cross-sectional study of 986 healthy Koreans. Three different case-control groups (i.e. G1, G2, and G3) were defined according to body mass index (BMI) and serum adiponectin levels. Allelic and genotypic associations of this gene with obesity were measured using multivariate logistic regression analyses in each group. RESULTS: The G allele of -11377C>G, a polymorphism located in the promoter region of the ADIPOQ gene (odds ratio (OR), 1.48; 95% confidence interval, 1.13-1.94) and most haplotypes including this allele significantly increased the risk for obesity. However, the OR decreased from 3.98 (G1 group) to 2.90 (G2 group) and 2.30 (G3 group) when a less strict definition of obesity was used. Most haplotypes, including this allele, significantly increased the risk of obesity. The statistical evidence from the GG genotype of -11377C>G (OR, 3.98) and the GT/GT diplotype composed of -11377G>C and +45T>G (OR, 5.20) confirmed the contribution of the G allele toward a predisposition for obesity. CONCLUSION: These results suggest the contribution of the ADIPOQ gene toward susceptibility to obesity in healthy Koreans. The high-risk genotypes and haplotypes identified here may provide more information for identifying individuals who are at risk of obesity.ope

Isolation and functional characterization of CE1 binding proteins

<p>Abstract</p> <p>Background</p> <p>Abscisic acid (ABA) is a plant hormone that controls seed germination, protective responses to various abiotic stresses and seed maturation. The ABA-dependent processes entail changes in gene expression. Numerous genes are regulated by ABA, and promoter analyses of the genes revealed that <it>cis</it>-elements sharing the ACGTGGC consensus sequence are ubiquitous among ABA-regulated gene promoters. The importance of the core sequence, which is generally known as ABA response element (ABRE), has been demonstrated by various experiments, and its cognate transcription factors known as ABFs/AREBs have been identified. Although necessary, ABRE alone is not sufficient, and another <it>cis</it>-element known as "coupling element (CE)" is required for full range ABA-regulation of gene expression. Several CEs are known. However, despite their importance, the cognate transcription factors mediating ABA response via CEs have not been reported to date. Here, we report the isolation of transcription factors that bind one of the coupling elements, CE1.</p> <p>Results</p> <p>To isolate CE1 binding proteins, we carried out yeast one-hybrid screens. Reporter genes containing a trimer of the CE1 element were prepared and introduced into a yeast strain. The yeast was transformed with library DNA that represents RNA isolated from ABA-treated Arabidopsis seedlings. From the screen of 3.6 million yeast transformants, we isolated 78 positive clones. Analysis of the clones revealed that a group of AP2/ERF domain proteins binds the CE1 element. We investigated their expression patterns and analyzed their overexpression lines to investigate the <it>in vivo </it>functions of the CE element binding factors (CEBFs). Here, we show that one of the CEBFs, AtERF13, confers ABA hypersensitivity in Arabidopsis, whereas two other CEBFs enhance sugar sensitivity.</p> <p>Conclusions</p> <p>Our results indicate that a group of AP2/ERF superfamily proteins interacts with CE1. Several CEBFs are known to mediate defense or abiotic stress response, but the physiological functions of other CEBFs remain to be determined. Our <it>in vivo </it>functional analysis of several CEBFs suggests that they are likely to be involved in ABA and/or sugar response. Together with previous results reported by others, our current data raise an interesting possibility that the coupling element CE1 may function not only as an ABRE but also as an element mediating biotic and abiotic stress responses.</p

Surface currents from hourly variations of suspended particulate matter from Geostationary Ocean Color Imager data

Surface currents in Korean coastal regions were obtained using the maximum cross-correlation method applied to hourly suspended particulate matter images from the Geostationary Ocean Color Imager. Preliminary current vectors were filtered out by applying a series of quality-control procedures. The current vectors resulting from the tests were compared with the currents from a numerical model with tide and wind field. It was found that the estimated currents were more similarly to the currents caused by both tide and wind. A high degree of discrepancy was detected in regions of strong tidal currents, where the fundamental assumption of horizontal movement was limited due to the dominant vertical tidal mixing in the shallow region. The hourly rotations of the current vectors within a day were clarified by a comparison of the time-varying orientation angles of tidal ellipses. This study emphasized how to understand the short-term surface flows from hourly high-resolution geostationary satellite images

Efficient hybrid organic-inorganic light emitting diodes with self-assembled dipole molecule deposited metal oxides

We investigate the effect of self-assembled dipole molecules (SADMs) on ZnO surface in hybrid organic-inorganic polymeric light-emitting diodes (HyPLEDs). Despite the SADM being extremely thin, the magnitude and orientation of SADM dipole moment effectively influenced the work function of the ZnO. As a consequence, the charge injection barrier between the conduction band of the ZnO and the lowest unoccupied molecular orbital of poly(9,9(')-dioctylfluorene)-co-benzothiadiazole could be efficiently controlled resulting that electron injection efficiency is remarkably enhanced. The HyPLEDs modified with a negative dipolar SADM exhibited enhanced device performances, which correspond to approximately a fourfold compared to those of unmodified HyPLEDs.open442

Eco-Innovation Indices as Tools for Measuring Eco-Innovation

Measuring eco-innovation helps us understand the overall trends and raises awareness in society. Measuring eco-innovation at the national level and making comparisons across countries may allow us to benchmark performance and foster policy learning. This paper assesses two indices developed in two different regions: The ASEM Eco-Innovation Index (ASEI) by the ASEM SMEs Eco-Innovation Center, based in Republic of Korea; and the Eco-Innovation Scoreboard (Eco-IS) developed by the Eco-Innovation Observatory, based in the European Union. This paper aims to examine and compare the features of both and attempts to obtain insights on their strengths and weaknesses. Towards this aim, our paper assesses those scoreboards against four criteria stemming from innovation analysis: (1) relevance of areas and stakeholders covered; (2) ability to indicate changes; (3) directions towards common goals; and (4) ability to facilitate further changes. We conclude both are promising, despite data shortages, and have great potential to contribute towards the sustainable development goals (SDGs), particularly with regard to the SDGs on sustainable industrialization and sustainable consumption and production. In comparison, the ASEI covers more countries than the Eco-IS. However, the ASEI has limitations on measuring indicators due to limited data availability in Asian countries. The Eco-IS is closely linked with the regional and national policies for eco-innovation in Europe, while the ASEI’s impact appears more limited, as of now. In conclusion, the research results give insights into key areas, goals and applications of eco-innovation indices, and can help upgrading eco-innovation indices. This research helps interpret the scores of two indices better and facilitate application of the scores in the multiple ways. It is expected that this research contributes to developing and modifying a global eco-innovation index and enhancing the ability of these indices to facilitate eco-innovation strategies at national levels and across relevant actors

Methano­ldinitrato[N-(2-pyridylmethyl­ene)aniline]copper(II)

The Cu atom in the title compound, [Cu(NO3)2(C12H10N2)(CH3OH)], adopts a square-pyramidal geometry, being ligated by two N atoms of the bidentate N-(2-pyridylmethyl­ene)­aniline (ppma) ligand, two O atoms of NO3 ligands and one O atom of a methanol molecule, which occupies the apical position. The phenyl ring on the ppma ligand is twisted out of the pyridine plane, forming a dihedral angle of 42.9 (1)°. In the crystal, inter­molecular O—H⋯O hydrogen bonds between methanol and NO3 ligands form an extensive one-dimensional network extending parallel to [100]

Enhanced Chromatin Accessibility and Recruitment of JUNB Mediate the Sustained IL-4 Expression in NFAT1 Deficient T Helper 2 Cells

Nuclear factor of activated T cells (NFAT) is a family of transcription factors composed of five proteins. Among them, NFAT1 is a predominant NFAT protein in CD4+ T cells. NFAT1 positively regulates transcription of a large number of inducible cytokine genes including IL-2, IL-4, IL-5 and other cytokines. However, disruption of NFAT1 results in an unexpected increase of IL-4. In this study, we have investigated the role of NFAT1 in regulation of IL-4 gene expression in T helper 2 cells (Th2) from an epigenetic viewpoint. NFAT1 deficient Th2 cells showed a sustained IL-4 expression while wild type (WT) cells reduced its expression. We tested whether epigenetic maintenance and changes in the chromatin architecture of IL-4 promoter locus play a role in differential IL-4 transcription between in WT and NFAT1 deficient Th2 cells. Compared with WT, NFAT1 deficient CD4+ Th2 cells exhibited enhanced chromatin accessibility with permissive histone modification and DNA demethylation in the IL-4 promoter region. Transcription factors bound to IL-4 promoter region in the absence of NFAT1 were identified by Micro-LC/LC-MS/MS analysis. Among the candidates, preferential recruitment of JUNB to the IL-4 promoter was confirmed by chromatin immunoprecipitation analysis. Overexpression of JUNB together with SATB1 synergistically upregulated IL-4 promoter activity, while knockdown JUNB significantly reduced IL-4 expression. Our results suggest that the prolonged IL-4 expression in NFAT1 deficient Th2 cells is mediated by preferential binding of JUNB/SATB1 to the IL-4 promoter with permissive chromatin architecture