Homeoprotein Hbx4 represses adhesion molecule governing cytokinesis and development

Homeobox genes encode proteins with a highly conserved DNA-binding motif and provoke morphological diversification of body segments by differentially controlling the expression of downstream targets. Here, we have identified _hbx4_, one of many homeobox genes in _Dictyostelium discoideum_ and investigated its role during growth and development. In suspension, Hbx4-overexpressing cells, Hbx4^OE^, showed defects in cytokinesis and growth rate. During development, Hbx4^OE^ and _hbx4_-disrupting cells, _hbx4¯_ made differences in shape of mound and slug, cell-type proportioning from wild type KAx3 cells. These phenotypes were similar to those of mutant defective in _cadA_ encoding Ca^2+^-dependent cell adhesion molecule so that we investigated the relationship between _hbx4_ and _cadA_. Overexpression of Hbx4 inhibited the expression of _cadA_ and cAMP also failed to stimulate _cadA_ in Hbx4^OE^. Furthermore, gel mobility shift assay showed the promoter of _cadA_ contained Hbx4-binding site, indicating Hbx4 negatively regulates the expression of _cadA_. Proteome analysis revealed that overexpression of Hbx4 repressed the _rdiA_ and _abpB_ encoding rho guanine nucleotide dissociation inhibitor1, RhoGDI1 and actin bundling protein 34, ABP34, respectively. And the overexpression of _cadA_ in Hbx4^OE^ cells rescued the defects and increased mRNA level of _rdiA_, _abpB_ and one of Rho GTPase, _rac1b_. These results suggested that Hbx4 can modulate cytokinesis, cell sorting and cell-type proportioning by repressing _cadA_ that regulates GTPase-dependent signaling pathway

Investigation of reliability of the cutoff probe by a comparison with Thomson scattering in high density processing plasmas

A “cutoff probe” uses microwaves to measure the electron density in a plasma. It is particularly attractive because it is easy to fabricate and use, its measurement is immune to surface contamination by dielectric materials, and it has a straightforward analysis to measure electron density in real time. In this work, we experimentally investigate the accuracy of the cutoff probe through a detailed comparison with Thomson scattering in a low temperature, high density processing plasma. The result shows that the electron density measured by the cutoff probe is lower than that by Thomson scattering and that the discrepancy of the two results becomes smaller as the gap between the two tips increases and/or the neutral gas pressure decreases. The underestimated electron density found by the cutoff probe can be explained by the influence of the probe holder, which becomes important as the pressure increases and the gap gets closer

Efficient and effective human action recognition in video through motion boundary description with a compact set of trajectories

Human action recognition (HAR) is at the core of human-computer interaction and video scene understanding. However, achieving effective HAR in an unconstrained environment is still a challenging task. To that end, trajectory-based video representations are currently widely used. Despite the promising levels of effectiveness achieved by these approaches, problems regarding computational complexity and the presence of redundant trajectories still need to be addressed in a satisfactory way. In this paper, we propose a method for trajectory rejection, reducing the number of redundant trajectories without degrading the effectiveness of HAR. Furthermore, to realize efficient optical flow estimation prior to trajectory extraction, we integrate a method for dynamic frame skipping. Experiments with four publicly available human action datasets show that the proposed approach outperforms state-of-the-art HAR approaches in terms of effectiveness, while simultaneously mitigating the computational complexity

Nanotextured Morphology of Poly(methyl methacrylate) and Ultraviolet Curable Poly(urethane acrylate) Blends via Phase Separation

Domain structures of spin-coated immiscible poly(methyl methacrylate) (PMMA) and ultraviolet (UV) curable poly(urethane acrylate) (PUA) blends were studied using atomic force microscopy (AFM). Spin casting the PMMA/PUA blends in propylene glycol monomethyl ether acetate (PGMEA) was accompanied with phase separation, and PUA was subsequently cross-linked under UV radiation. Selective dissolution of PMMA in the phase-separated films was feasible using tetrahydrofuran (THF) solvent after the UV curing process, because the cured PUA material is highly stable against THF. Morphology of phase-separated structure, including domain size and height, could be controlled by varying total concentration of the blended solution, and various nanoscale features such as island-like and hole-like structures were achieved by changing weight ratio of the two immiscible polymers

Effects of Radix Adenophorae and Cyclosporine A on an OVA-Induced Murine Model of Asthma by Suppressing to T Cells Activity, Eosinophilia, and Bronchial Hyperresponsiveness

The present study is performed to investigate the inhibitory effects of Radix Adenophorae extract (RAE) on ovalbumin-induced asthma murine model. To study the anti-inflammatory and antiasthmatic effects of RAE, we examined the development of pulmonary eosinophilic inflammation and inhibitory effects of T cells in murine by RAE and cyclosporine A (CsA). We examined determination of airway hyperresponsiveness, flow cytometric analysis (FACS), enzyme-linked immunosorbent assay (ELISA), quantitative real time (PCR), hematoxylin-eosin staining, and Masson trichrome staining in lung tissue, lung weight, total cells, and eosinophil numbers in lung tissue. We demonstrated how RAE suppressed development on inflammation and decreased airway damage

Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine

Background: Mapping of tissue structure at the cellular, circuit, and organ-wide scale is important for understanding physiological and biological functions. A bio-electrochemical technique known as CLARITY used for three-dimensional anatomical and phenotypical mapping within transparent intact tissues has been recently developed. This method provided a major advance in understanding the structure-function relationships in circuits of the nervous system and organs by using whole-body clearing. Thus, in the present study, we aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs. Results: We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images. We also determined the appropriate replacement cycles of clearing solution for each type of organ, and convincingly demonstrated that 250–280 mA is the ideal range of electrical current for tissue clearing. We then acquired each type of cleared organs including brain, pancreas, liver, lung, kidney, and intestine. Additionally, we determined the images of axon fibers of hippocampal region, the Purkinje layer of cerebellum, and vessels and cellular nuclei of pancreas. Conclusions: CLARITY is an innovative biochemical technology for the structural and molecular analysis of various types of tissue. We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type. These optimized methods will be useful for the application of CLARITY to various types of organs. Electronic supplementary material The online version of this article (doi:10.1186/s12861-014-0048-3) contains supplementary material, which is available to authorized users

Isolation of human Dna2 endonuclease and characterization of its enzymatic properties

In eukaryotes, the creation of ligatable nicks in DNA from flap structures generated by DNA polymerase δ-catalyzed displacement DNA synthesis during Okazaki fragment processing depends on the combined action of Fen1 and Dna2. These two enzymes contain partially overlapping but distinct endonuclease activities. Dna2 is well-suited to process long flaps, which are converted to nicks by the subsequent action of Fen1. In this report, we purified human Dna2 as a recombinant protein from human cells transfected with the cDNA of the human homologue of Saccharomyces cerevisiae Dna2. We demonstrated that the purified human Dna2 enzyme contains intrinsic endonuclease and DNA-dependent ATPase activities, but is devoid of detectable DNA helicase activity. We determined a number of enzymatic properties of human Dna2 including its substrate specificity. When both 5′ and 3′ tailed ssDNAs were present in a substrate, such as a forked-structured one, both single-stranded regions were cleaved by human Dna2 (hDna2) with equal efficiency. Based on this and other properties of hDna2, it is likely that this enzyme facilitates the removal of 5′ and 3′ regions in equilibrating flaps that are likely to arise during the processing of Okazaki fragments in human cells