Insulin direct pancreatic progenitor cell differentiation via Pdx1 regulation

poster abstractDifferentiation of early foregut endoderm into pancreatic endocrine and exocrine cells depends on a sequence of gene expression directed by various signals secreted from nearby tissue. Prior studies have shown that the pancreas is derived from Pdx1+ progenitor cells; however Pdx1 is turned off in pancreatic exocrine cells and α cells while maintained in β cells. Here, using zebrafish genetic knockdown, we showed that insulin secreted by early β cells can repress Pdx1 expression in pancreatic progenitor cells allowing them to differentiate to different pancreatic cell types. Knockdown of insulin gene severely impairs exocrine pancreas development. My results further demonstrate that inhibition of insulin signaling can induce pre-differentiation of Pdx1+ progenitor cells to β cells and Pdx1+ α cells. These Pdx1+ α cells can transdifferentiate to β cells following β cell ablation. Overall, these data represent the first in vivo evidence of local insulin signaling on pancreas development via regulation of Pdx1 expression

Highly selective and sensitive macrocycle-based dinuclear foldamer for fluorometric and colorimetric sensing of citrate in water.

The selective detection of citrate anions is essential for various biological functions in living systems. A quantitative assessment of citrate is required for the diagnosis of various diseases in the human body; however, it is extremely challenging to develop efficient fluorescence and color-detecting molecular probes for sensing citrate in water. Herein, we report a macrocycle-based dinuclear foldamer (1) assembled with eosin Y (EY) that has been studied for anion binding by fluorescence and colorimetric techniques in water at neutral pH. Results from the fluorescence titrations reveal that the 1·EY ensemble strongly binds citrate anions, showing remarkable selectivity over a wide range of inorganic and carboxylate anions. The addition of citrate anions to the 1·EY adduct led to a large fluorescence enhancement, displaying a detectable color change under both visible and UV light in water up to 2 μmol. The biocompatibility of 1·EY as an intracellular carrier in a biological system was evaluated on primary human foreskin fibroblast (HF) cells, showing an excellent cell viability. The strong binding properties of the ensemble allow it to be used as a highly sensitive, detective probe for biologically relevant citrate anions in various applications

Retrieval of Raindrop Size Distribution, Vertical Air Velocity and Water Vapor Attenuation Using Dual-Wavelength Doppler Radar Observations

Two techniques for retrieving the slope and intercept parameters of an assumed exponential raindrop size distribution (RSD), vertical air velocity, and attenuation by precipitation and water vapor in light stratiform rain using observations by airborne, nadir looking dual-wavelength (X-band, 3.2 cm and W-band, 3.2 mm) radars are presented. In both techniques, the slope parameter of the RSD and the vertical air velocity are retrieved using only the mean Doppler velocities at the two wavelengths. In the first method, the intercept of the RSD is estimated from the observed reflectivity at the longer wavelength assuming no attenuation at that wavelength. The attenuation of the shorter wavelength radiation by precipitation and water vapor are retrieved using the observed reflectivity at the shorter wavelength. In the second technique, it is assumed that the longer wavelength suffers attenuation only in the melting band. Then, assuming a distribution of water vapor, the melting band attenuation at both wavelengths and the rain attenuation at the shorter wavelength are retrieved. Results of the retrievals are discussed and several physically meaningful results are presented

Identification of common blood gene signatures for the diagnosis of renal and cardiac acute allograft rejection.

To test, whether 10 genes, diagnostic of renal allograft rejection in blood, are able to diagnose and predict cardiac allograft rejection, we analyzed 250 blood samples from heart transplant recipients with and without acute rejection (AR) and with cytomegalovirus (CMV) infection by QPCR. A QPCR-based logistic regression model was built on 5 of these 10 genes (AR threshold composite score >37%  = AR) and tested for AR prediction in an independent set of 109 samples, where it correctly diagnosed AR with 89% accuracy, with no misclassifications for AR ISHLT grade 1b. CMV infection did not confound the AR score. The genes correctly diagnosed AR in a blood sample within 6 months prior to biopsy diagnosis with 80% sensitivity and untreated grade 1b AR episodes had persistently elevated scores until 6 months after biopsy diagnosis. The gene score was also correlated with presence or absence of cardiac allograft vasculopathy (CAV) irrespective of rejection grade. In conclusion, there is a common transcriptional axis of immunological trafficking in peripheral blood in both renal and cardiac organ transplant rejection, across a diverse recipient age range. A common gene signature, initially identified in the setting of renal transplant rejection, can be utilized serially after cardiac transplantation, to diagnose and predict biopsy confirmed acute heart transplant rejection

An insulin signaling feedback loop regulates pancreas progenitor cell differentiation during islet development and regeneration

As one of the key nutrient sensors, insulin signaling plays an important role in integrating environmental energy cues with organism growth. In adult organisms, relative insufficiency of insulin signaling induces compensatory expansion of insulin-secreting pancreatic beta (β) cells. However, little is known about how insulin signaling feedback might influence neogenesis of β cells during embryonic development. Using genetic approaches and a unique cell transplantation system in developing zebrafish, we have uncovered a novel role for insulin signaling in the negative regulation of pancreatic progenitor cell differentiation. Blocking insulin signaling in the pancreatic progenitors hastened the expression of the essential β cell genes insulin and pdx1, and promoted β cell fate at the expense of alpha cell fate. In addition, loss of insulin signaling promoted β cell regeneration and destabilization of alpha cell character. These data indicate that insulin signaling constitutes a tunable mechanism for β cell compensatory plasticity during early development. Moreover, using a novel blastomere-to-larva transplantation strategy, we found that loss of insulin signaling in endoderm-committed blastomeres drove their differentiation into β cells. Furthermore, the extent of this differentiation was dependent on the function of the β cell mass in the host. Altogether, our results indicate that modulation of insulin signaling will be crucial for the development of β cell restoration therapies for diabetics; further clarification of the mechanisms of insulin signaling in β cell progenitors will reveal therapeutic targets for both in vivo and in vitro β cell generation

Facial expression cloning optimization method based Laplace operator.

In view of the reality of facial expression cloning and efficiency of expression reconstruction, a novel method based on motion capture data is proposed. After capturing the data of six fundamental expressions, it normalizes these data to make them in the same range. Then 41 points are chosen in critical areas of facial expression and it gets cloning expression using Laplace deformation algorithm with convex weight which can preserve the details of facial expression to avoid the low fidelity of uniform weights and unstable calculation of cotangent weights. Experimental results show that this method can generate realistic and natural expression animations and the efficiency of facial expression cloning is improved significantly

Retrieve Optically Thick Ice Cloud Microphysical Properties by Using Airborne Dual-Wavelength Radar Measurements

An algorithm to retrieve optically thick ice cloud microphysical property profiles is developed by using the GSFC 9.6 GHz ER-2 Doppler Radar (EDOP) and the 94 GHz Cloud Radar System (CRS) measurements aboard the high-altitude ER-2 aircraft. In situ size distribution and total water content data from the CRYSTAL-FACE field campaign are used for the algorithm development. To reduce uncertainty in calculated radar reflectivity factors (Ze) at these wavelengths, coincident radar measurements and size distribution data are used to guide the selection of mass-length relationships and to deal with the density and non-spherical effects of ice crystals on the Ze calculations. The algorithm is able to retrieve microphysical property profiles of optically thick ice clouds, such as, deep convective and anvil clouds, which are very challenging for single frequency radar and lidar. Examples of retrieved microphysical properties for a deep convective clouds are presented, which show that EDOP and CRS measurements provide rich information to study cloud structure and evolution. Good agreement between IWPs derived from an independent submillimeter-wave radiometer, CoSSIR, and dual-wavelength radar measurements indicates accuracy of the IWC retrieved from the two-frequency radar algorithm

A Novel Role of Protein Tyrosine Kinase2 in Mediating Chloride Secretion in Human Airway Epithelial Cells

Ca2+ activated Cl− channels (CaCC) are up-regulated in cystic fibrosis (CF) airway surface epithelia. The presence and functional properties of CaCC make it a possible therapeutic target to compensate for the deficiency of Cl− secretion in CF epithelia. CaCC is activated by an increase in cytosolic Ca2+, which not only activates epithelial CaCCs, but also inhibits epithelial Na+ hyperabsorption, which may also be beneficial in CF. Our previous study has shown that spiperone, a known antipsychotic drug, activates CaCCs and stimulates Cl− secretion in polarized human non-CF and CF airway epithelial cell monolayers in vitro, and in Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) knockout mice in vivo. Spiperone activates CaCC not by acting in its well-known role as an antagonist of either 5-HT2 or D2 receptors, but through a protein tyrosine kinase-coupled phospholipase C-dependent pathway. Moreover, spiperone independently activates CFTR through a novel mechanism. Herein, we performed a mass spectrometry analysis and identified the signaling molecule that mediates the spiperone effect in activating chloride secretion through CaCC and CFTR. Proline-rich tyrosine kinase 2 (PYK2) is a non-receptor protein tyrosine kinase, which belongs to the focal adhesion kinase family. The inhibition of PYK2 notably reduced the ability of spiperone to increase intracellular Ca2+ and Cl− secretion. In conclusion, we have identified the tyrosine kinase, PYK2, as the modulator, which plays a crucial role in the activation of CaCC and CFTR by spiperone. The identification of this novel role of PYK2 reveals a new signaling pathway in human airway epithelial cells

Glucagon is essential for alpha cell transdifferentiation and beta cell neogenesis

The interconversion of cell lineages via transdifferentiation is an adaptive mode of tissue regeneration and an appealing therapeutic target. However, its clinical exploitation is contingent upon the discovery of contextual regulators of cell fate acquisition and maintenance. In murine models of diabetes, glucagon-secreting alpha cells transdifferentiate into insulin-secreting beta cells following targeted beta cell depletion, regenerating the form and function of the pancreatic islet. However, the molecular triggers of this mode of regeneration are unknown. Here, using lineage-tracing assays in a transgenic zebrafish model of beta cell ablation, we demonstrate conserved plasticity of alpha cells during islet regeneration. In addition, we show that glucagon expression is upregulated after injury. Through gene knockdown and rescue approaches, we also find that peptides derived from the glucagon gene are necessary for alpha-to-beta cell fate switching. Importantly, whereas beta cell neogenesis was stimulated by glucose, alpha-to-beta cell conversion was not, suggesting that transdifferentiation is not mediated by glucagon/GLP-1 control of hepatic glucose production. Overall, this study supports the hypothesis that alpha cells are an endogenous reservoir of potential new beta cells. It further reveals that glucagon plays an important role in maintaining endocrine cell homeostasis through feedback mechanisms that govern cell fate stability

Multifunctional three-dimensional macroporous nanoelectronic networks for smart materials

Seamless and minimally-invasive integration of three-dimensional (3D) electronic circuitry within host materials could enable the development of materials systems that are self- monitoring and allow for communication with external environments. Here, we report a general strategy for preparing ordered 3D interconnected and addressable macroporous nanoelectronic networks from ordered two-dimensional (2D) nanowire nanoelectronic “precursors”, which are fabricated by conventional lithography. The 3D networks have porosities larger than 99%, contain ca. 100’s of addressable nanowire devices, and have feature sizes from the 10 micron scale (for electrical and structural interconnections) to the 10 nanometer scale (for device elements). The macroporous nanoelectronic networks were merged with organic gels and polymers to form hybrid materials in which the basic physical and chemical properties of the host were not substantially altered, and electrical measurements further show a > 90% yield of active devices in the hybrid materials. The positions of the nanowire devices were located within 3D hybrid materials with ca. 14 nm resolution through simultaneous nanowire device photocurrent/confocal microscopy imaging measurements. In addition, we explored functional properties of these hybrid materials, including (i) mapping time-dependent pH changes throughout a nanowire network/agarose gel sample during external solution pH changes, and (ii) characterizing the strain field in a hybrid nanoelectronic elastomer structures subject to uniaxial and bending forces. The seamless incorporation of active nanoelectronic networks within 3D materials opens up a powerful approach to smart materials in which the capabilities of multi- functional nanoelectronics allow for active monitoring and control of host systems.Chemistry and Chemical BiologyEngineering and Applied Science