Size-scaling effects for microparticles and cells manipulated by optoelectronic tweezers

In this work, we investigated the use of optoelectronic tweezers (OET) to manipulate objects that are larger than those commonly positioned with standard optical tweezers. We studied the forces that could be produced on differently sized polystyrene microbeads and MCF-7 breast cancer cells with light-induced dielectrophoresis (DEP). It was found that the DEP force imposed on the bead/cell did not increase linearly with the volume of the bead/cell, primarily because of the non-uniform distribution of the electric field above the OET bottom plate. Although this size-scaling work focuses on microparticles and cells, we propose that the physical mechanism elucidated in this research will be insightful for other micro-objects, biological samples, and micro-actuators undergoing OET manipulation

Manufacturing with light – micro-assembly of opto-electronic microstructures

Optical micromanipulation allows the movement and patterning of discrete micro-particles within a liquid environment. However, for manufacturing applications it is desirable to remove the liquid, leaving the patterned particles in place. In this work, we have demonstrated the use of optoelectronic tweezers (OET) to manipulate and accurately assemble Sn62Pb36Ag2 solder microspheres into tailored patterns. A technique based on freeze-drying technology was then developed that allows the assembled patterns to be well preserved and fixed in place after the liquid medium in the OET device is removed. After removing the liquid from the OET device and subsequently heating the assembled pattern and melting the solder microspheres, electrical connections between the microspheres were formed, creating a permanent conductive bridge between two isolated metal electrodes. Although this method is demonstrated with 40 µm diameter solder beads arranged with OET, it could be applied to a great range of discrete components from nanowires to optoelectronic devices, thus overcoming one of the basic hurdles in using optical micromanipulation techniques in a manufacturing micro-assembly setting

Ankyrin repeat and SOCS box containing protein 4 (Asb-4) colocalizes with insulin receptor substrate 4 (IRS4) in the hypothalamic neurons and mediates IRS4 degradation

Abstract Background The arcuate nucleus of the hypothalamus regulates food intake. Ankyrin repeat and SOCS box containing protein 4 (Asb-4) is expressed in neuropeptide Y and proopiomelanocortin (POMC) neurons in the arcuate nucleus, target neurons in the regulation of food intake and metabolism by insulin and leptin. However, the target protein(s) of Asb-4 in these neurons remains unknown. Insulin receptor substrate 4 (IRS4) is an adaptor molecule involved in the signal transduction by both insulin and leptin. In the present study we examined the colocalization and interaction of Asb-4 with IRS4 and the involvement of Asb-4 in insulin signaling. Results In situ hybridization showed that the expression pattern of Asb-4 was consistent with that of IRS4 in the rat brain. Double in situ hybridization showed that IRS4 colocalized with Asb-4, and both Asb-4 and IRS4 mRNA were expressed in proopiomelanocortin (POMC) and neuropeptide Y (NPY) neurons within the arcuate nucleus of the hypothalamus. In HEK293 cells co-transfected with Myc-tagged Asb-4 and Flag-tagged IRS4, Asb-4 co-immunoprecipitated with IRS4; In these cells endogenous IRS4 also co-immunoprecipitated with transfected Myc-Asb-4; Furthermore, Asb-4 co-immunoprecipitated with IRS4 in rat hypothalamic extracts. In HEK293 cells over expression of Asb-4 decreased IRS4 protein levels and deletion of the SOCS box abolished this effect. Asb-4 increased the ubiquitination of IRS4; Deletion of SOCS box abolished this effect. Expression of Asb-4 decreased both basal and insulin-stimulated phosphorylation of AKT at Thr308. Conclusions These data demonstrated that Asb-4 co-localizes and interacts with IRS4 in hypothalamic neurons. The interaction of Asb-4 with IRS4 in cell lines mediates the degradation of IRS4 and decreases insulin signaling

Modulation of food intake by mTOR signalling in the dorsal motor nucleus of the vagus in male rats: focus on ghrelin and nesfatin‐1

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/101864/1/expphysiol.2013.074930.pd

Mutations in KEOPS-Complex Genes Cause Nephrotic Syndrome with Primary Microcephaly

Galloway-Mowat syndrome (GAMOS) is an autosomal-recessive disease characterized by the combination of early-onset nephrotic syndrome (SRNS) and microcephaly with brain anomalies. Here we identified recessive mutations in OSGEP, TP53RK, TPRKB, and LAGE3, genes encoding the four subunits of the KEOPS complex, in 37 individuals from 32 families with GAMOS. CRISPR-Cas9 knockout in zebrafish and mice recapitulated the human phenotype of primary microcephaly and resulted in early lethality. Knockdown of OSGEP, TP53RK, or TPRKB inhibited cell proliferation, which human mutations did not rescue. Furthermore, knockdown of these genes impaired protein translation, caused endoplasmic reticulum stress, activated DNA-damage-response signaling, and ultimately induced apoptosis. Knockdown of OSGEP or TP53RK induced defects in the actin cytoskeleton and decreased the migration rate of human podocytes, an established intermediate phenotype of SRNS. We thus identified four new monogenic causes of GAMOS, describe a link between KEOPS function and human disease, and delineate potential pathogenic mechanisms

Morphology and rheological properties of silica-​filled poly(carbonate)​/poly(methyl methacrylate) blends

The effect of silica nanoparticles on the morphol. and the rheol. properties was investigated in the immiscible polymer blend poly(carbonate)¿/poly(Me methacrylate) (PC¿/PMMA)¿. In the melt state, the linear viscoelastic properties of the nanocomposite showed a redn. effect of the silica nanoparticles on the mobility of one of the polymer which is related to the state of distribution of the silica nanoparticles. Hydrophilic and hydrophobic silica particles were used to study particle migration and their effects on the morphol. and it was shown that the distribution of the nanoparticles depends on the balance of interactions between the surface of the particles and the polymer components. The effect on the coarsening kinetics was investigated in both hydrophilic and hydrophobic silica-¿filled blends. Compared to the hydrophilic silica, a better compatibilization can be obtained by introducing the hydrophobic silica particles at the PC¿/PMMA interface as the solid barrier

mTOR-dependent Modulation of Gastric Nesfatin-1/NUCB2

Background: Nesfatin-1, an 82 amino acid peptide derived from the prohormone nucleobindin-2 (NUCB2), is a novel satiety hormone acting through a leptin-independent mechanism in the hypothalamus. The mechanisms by which production of nesfatin-1/NUCB2 is regulated remain unknown. Methods: Nesfatin-1/NUCB2 mRNA and immunoreactivity were examined in gastric tissue and Min-6 cells by RT-PCR and immunofluorescent staining or Western blotting. Results: Nesfatin-1/NUCB2 is co-localized with pS6K1, the downstream target of mammalian target of rapamycin (mTOR), in gastric X/A like cells. A parallel relationship between gastric mTOR signaling and nesfatin-1/NUCB2 was observed during changes in energy status. Both mTOR activity and gastric nesfatin-1/NUCB2 were down-regulated by fasting, and returned to basal levels with re-feeding. In high fat diet induced obese mice, gastric mTOR signaling and nesfatin-1/NUCB2 were increased. Inhibition of the gastric mTOR signaling by rapamycin attenuated the expression of gastric nesfatin-1/NUCB2 mRNA and protein in both lean and obese mice. Attenuation of mTOR activity by rapamycin or over-expression of TSC1 or TSC2 reduced the expression of nesfatin-1/NUCB2 in Min-6 cells, suggesting a direct effect of mTOR signaling. Conclusion: Gastric mTOR is a gastric energy sensor whose activity is linked to the regulation of gastric nesfatin-1/NUCB2. Copyright (C) 2012 S. Karger AG, Baselhttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000302752600018&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701Cell BiologyPhysiologySCI(E)PubMed19ARTICLE3-4493-5002

Evidence of Intrinsic Double Acceptor in GaAs

Acceptors present in undoped p‐type conducting GaAs have been studied with photoluminescence, temperature‐dependent Hall measurements, deep level transient spectroscopy, and spark source mass spectrometry. It is shown that p‐type conduction is due to presence of the shallow acceptor CAs and the cation antisite double acceptor GaAs. The first and second ionization energies determined for GaAs are 77 and 230 meV from the valence‐band edge