Sprouty4 Is an Endogenous Negative Modulator of TrkA Signaling and Neuronal Differentiation Induced by NGF

The Sprouty (Spry) family of proteins represents endogenous regulators of downstream signaling pathways induced by receptor tyrosine kinases (RTKs). Using real time PCR, we detect a significant increase in the expression of Spry4 mRNA in response to NGF, indicating that Spry4 could modulate intracellular signaling pathways and biological processes induced by NGF and its receptor TrkA. In this work, we demonstrate that overexpression of wild-type Spry4 causes a significant reduction in MAPK and Rac1 activation and neurite outgrowth induced by NGF. At molecular level, our findings indicate that ectopic expression of a mutated form of Spry4 (Y53A), in which a conserved tyrosine residue was replaced, fail to block both TrkA-mediated Erk/MAPK activation and neurite outgrowth induced by NGF, suggesting that an intact tyrosine 53 site is required for the inhibitory effect of Spry4 on NGF signaling. Downregulation of Spry4 using small interference RNA knockdown experiments potentiates PC12 cell differentiation and MAPK activation in response to NGF. Together, these findings establish a new physiological mechanism through which Spry4 regulates neurite outgrowth reducing not only the MAPK pathway but also restricting Rac1 activation in response to NGF

Thermal modelling of gas generation and retention in the Jurassic organic-rich intervals in the Darquain field, Abadan Plain, SW Iran

The petroleum system with Jurassic source rocks is an important part of the hydrocarbons discovered in the Middle East. Limited studies have been done on the Jurassic intervals in the 26,500 km2 Abadan Plain in south-west Iran, mainly due to the deep burial and a limited number of wells that reach the basal Jurassic successions. The goal of this study was to evaluate the Jurassic organic-rich intervals and shale gas play in the Darquain field using organic geochemistry, organic petrography, biomarker analysis, and basin modelling methods. This study showed that organic-rich zones present in the Jurassic intervals of Darquain field could be sources of conventional and unconventional gas reserves. The organic matter content of samples from the organic-rich zones corresponds to medium-to-high-sulphur kerogen Type II-S marine origin. The biomarker characteristics of organic-rich zones indicate carbonate source rocks that contain marine organic matter. The biomarker results also suggest a marine environment with reducing conditions for the source rocks. The constructed thermal model for four pseudo-wells indicates that, in the kitchen area of the Jurassic gas reserve, methane has been generated in the Sargelu and Neyriz source rocks from Early Cretaceous to recent times and the transformation ratio of organic matter is more than 97%. These organic-rich zones with high initial total organic carbon (TOC) are in the gas maturity stage [1.5–2.2% vitrinite reflectance in oil (Ro)] and could be good unconventional gas reserves and gas source rocks. The model also indicates that there is a huge quantity of retained gas within the Jurassic organic-rich intervals

GABA Maintains the Proliferation of Progenitors in the Developing Chick Ciliary Marginal Zone and Non-Pigmented Ciliary Epithelium

GABA is more than the main inhibitory neurotransmitter found in the adult CNS. Several studies have shown that GABA regulates the proliferation of progenitor and stem cells. This work examined the effects of the GABAA receptor system on the proliferation of retinal progenitors and non-pigmented ciliary epithelial (NPE) cells. qRT-PCR and whole-cell patch-clamp electrophysiology were used to characterize the GABAA receptor system. To quantify the effects on proliferation by GABAA receptor agonists and antagonists, incorporation of thymidine analogues was used. The results showed that the NPE cells express functional extrasynaptic GABAA receptors with tonic properties and that low concentration of GABA is required for a baseline level of proliferation. Antagonists of the GABAA receptors decreased the proliferation of dissociated E12 NPE cells. Bicuculline also had effects on progenitor cell proliferation in intact E8 and E12 developing retina. The NPE cells had low levels of the Cl–transporter KCC2 compared to the mature retina, suggesting a depolarising role for the GABAA receptors. Treatment with KCl, which is known to depolarise membranes, prevented some of the decreased proliferation caused by inhibition of the GABAA receptors. This supported the depolarising role for the GABAA receptors. Inhibition of L-type voltage-gated Ca2+ channels (VGCCs) reduced the proliferation in the same way as inhibition of the GABAA receptors. Inhibition of the channels increased the expression of the cyclin-dependent kinase inhibitor p27KIP1, along with the reduced proliferation. These results are consistent with that when the membrane potential indirectly regulates cell proliferation with hyperpolarisation of the membrane potential resulting in decreased cell division. The increased expression of p27KIP1 after inhibition of either the GABAA receptors or the L-type VGCCs suggests a link between the GABAA receptors, membrane potential, and intracellular Ca2+ in regulating the cell cycle

Hawking Radiation from Higher-Dimensional Black Holes

We review the quantum field theory description of Hawking radiation from evaporating black holes and summarize what is known about Hawking radiation from black holes in more than four space-time dimensions. In the context of the Large Extra Dimensions scenario, we present the theoretical formalism for all types of emitted fields and a selection of results on the radiation spectra. A detailed analysis of the Hawking fluxes in this case is essential for modelling the evaporation of higher-dimensional black holes at the LHC, whose creation is predicted by low-energy models of quantum gravity. We discuss the status of the quest for black-hole solutions in the context of the Randall-Sundrum brane-world model and, in the absence of an exact metric, we review what is known about Hawking radiation from such black holes

MITOCHONDRIAL BIOGENESIS AFTER REPEATED BOUTS OF DISUSE

Jacob L. Brown1, Richard A. Perry, Jr1., Kevin L. Shimkus2,David E. Lee1, Megan E. Rosa1, Jessica M. Cardin2, Lemuel A. Brown1, Elizabeth K. McBee1, Yasaman Sharazi-Fard2, Harry Hogan2, James D. Fluckey2, Tyrone A. Washington1 & Nicholas P.Greene1,2 1University of Arkansas, Fayetteville, Arkansas. 2Texas A&M University, College Station, Texas Muscular disuse affects a great number of people have sedentary lifestyles and/or chronic disease. Disuse has been shown to cause severe muscular atrophy and to disrupt mitochondrial quality. PURPOSE: To examine if disuse affects mitochondrial biogenesis, and if resistance exercise following bouts of unloading can promote biogenesis. METHODS: Sprague-Dawley rats were subjected to chronic disuse atrophy by hindlimb unloading (28-d, 1HU) followed by ambulatory recovery (56-d) with (1HU+EX) and without (1HU+REC) resistance exercise. To mimic repeated bouts of disuse animals were subjected to a second bout of HU (28-d, 2HU) and again allowed ambulatory recovery with (2HU+EX) or without (2HU+REC) resistance exercise. Control (CON) animals were allowed normal cage activity throughout. Samples were analyzed for Pgc-1α, Tfam, Nrf2 and Pparα gene expression by real time RT-PCR. To test if disuse impacted mitochondrial biogenesis regulators a T-Test was performed between CON and 1HU groups, to test impact of reloading and exercise data were analyzed by one-way ANOVA across all groups with α set at PRESULTS: Pgc-1α expression decreased by 59% (p=0.042) and Nrf2 by 75% (p=0.047) following disuse (1HU) compared to CON. 1HU+Ex showed a 280% increase in Pparα expression (p=0.005) as well as a 278% increase in Tfam expression (p=0.013) compared to CON. Pgc-1α, Pparα, and Tfam displayed a greater increase in expression with exercise recovery (1HU+Ex) than without (1HU+Rec). Pgc-1α showed an 80% increase in expression (p=0.05), Pparα showed a 208% increase in expression (p=0.01), and Tfam showed a 195% increase in expression (p=0.01) when comparing 1HU+Ex and 1HU+Rec. Nrf2 decreased by 61% (p=0.008) with 2HU. Expression of other biogenesis markers was not changed in the 2HU group. Neither 2HU+Ex nor 2HU+Rec were able to attenuate the loss of Nrf2 expression. CONCLUSION: A single bout of disuse significantly decreases the expression of Pgc-1α and Nrf2. 1HU+Ex promotes mitochondrial biogenesis more than 1HU+Rec. Multiple bouts of disuse decreases the expression of Nrf2. 2HU+Ex and 2HU+Rec does not attenuate the loss of Nrf2 expression. More research needs to be conducted to examine other aspects of mitochondrial quality such as mitochondrial dynamics and autophagy. Funded By National Space Biomedical Research Institut

AUTOPHAGY FOLLOWING MULTIPLE BOUTS OF HINDLIMB UNLOADING

Elizabeth K. McBee1, Jacob L. Brown1, Richard A. Perry, Jr1, Kevin L. Shimkus2,David E. Lee1, Megan E. Rosa1, Jessica M. Cardin2, Lemuel A. Brown1, Yasaman Shirazi-Fard2, Harry A. Hogan2, James D. Fluckey2, Tyrone A. Washington1, Nicholas P. Greene1,2 1University of Arkansas, Fayetteville, AR. 2Texas A&M University, College Station, TX; e-mail: [email protected] Muscular disuse affects an abundance of people with sedentary lifestyles and/or chronic diseases. Disuse has proven to cause severe muscular atrophy and disrupt mitochondrial quality. Autophagy is a cellular mechanism used to remove wasteful or damaged materials, mitophagy is the process of autophagically removing presumably damaged mitochondria. Beclin1 and Atg7 are machinery for autophagy, while Bnip3 specifically regulates mitophagy. Any dysregulation of autophagic processes may significantly impair cellular health following muscle disuse. PURPOSE: The purpose of this study was to investigate whether multiple bouts of muscle disuse (hindlimb unloading, HU) affect the expression of markers for autophagy. METHODS: Sprague-Dawley rats were subjected to chronic disuse atrophy by hindlimb unloading (28-d, 1HU) followed by ambulatory recovery (56-d) (1HU+REC). To mimic repeated bouts of disuse, the animals were subjected to a second bout of HU (28-d, 2HU) and then allowed ambulatory recovery (2HU+REC). Control (CON) animals were allowed normal cage activity throughout. Samples were analyzed for Beclin1, Atg7, and Bnip3 mRNA content by real time RT-PCR. To verify if HU impacted autophagy markers pre-planned T-tests were performed comparing CON and 1HU. To test if multiple bouts of disuse and reloading impacted regulators of autophagy, a one-way ANOVA across all groups was employed with α set at p\u3c0.05. RESULTS: Beclin1, Atg7, and Bnip3 mRNA contents were not different, 270% greater, and ~50% lower following 1HU compared to CON, respectively. Across all conditions, Beclin1 was unchanged. Atg7 was not different from CON in 1HU+REC, 2HU, and 2HU+REC groups. Atg7 was ~30% of 1HU in 1HU+REC and 2HU groups. Bnip3 showed no further differences among conditions. CONCLUSION: A single bout of HU enhanced the capacity for general autophagy, while diminishing the capacity for mitophagy. Recovery from repeated bouts of HU did not alter mRNA levels of autophagic markers relative to CON. These data show that autophagic markers are dysregulated with HU, and that animals adapted to multiple bouts of HU, which dampened the impact of subsequent HU on autophagic markers. Funding provided by NASA Grant Number NNX08AQ35

Inspection methods for 3D concrete printing

3D Concrete Printing (3DCP) is being used for off-site manufacture of many elements found in the built environment, ranging from furniture to bridges. The advantage of these methods is the value added through greater geometrical freedom because a mould is not needed to create the form. In recent years, research has focused on material properties both in the wet and hardened state, while less attention has been paid to verifying printed forms through geometry measurement. Checking conformity is a critical aspect of manufacturing quality control, particularly when assembling many components, or when integrating/interfacing parts into/with existing construction. This paper takes a case study approach to explore applications of digital measurement systems prior to, during, after manufacture using 3DCP and after the assembly of a set of 3DCP parts and discusses the future prospects for such technology as part of geometry quality control for the procurement of 3DCP elements for the built environment