Peptide Expression and Purification to Elucidate the Mechanism of Vascular Alpha2C-Adrenoceptor Traslocation

In response to physiological cold temperatures, cutaneous blood vessels constrict to reduce heat loss. This constriction is mediated by smooth muscle G protein-coupled α2C-adrenoceptors (α2C-ARs), which translocate from the trans-Golgi to the plasma membrane upon cooling. The cellular mechanisms involved in receptor translocation are not fully understood; however, this translocation is hypothesized to be dependent upon a protein-protein interaction between the α2C-AR C-terminus and the protein filamin-2. This portion of the receptor translocation mechanism can be tested experimentally by decoy peptide interference. The decoy peptide contains a TAT domain and an identical C-terminus to the α2C-AR, allowing the peptide to penetrate the cell membrane and mimic the α2C-AR, respectively. We predict that interfering with this interaction will inhibit translocation of α2C-ARs and subsequent receptor function. Therefore, the goal of this study is to generate and purify a decoy peptide, confirm delivery to human VSM cells, and assess cellular activity for endogenous α2C-ARs. The DNA sequence encoding 11 amino acids of the cell-permeable HIV transactivator of transduction (TAT) domain was fused in frame with the α2C-AR C-terminus to generate the p-TAT-α2C-AR-C-terminus bacterial expression vector. BL21-CodonPlus competent cells were used to optimize peptide expression at 30°C. The parameters tested during bacterial growth included the presence of antibiotics, induction time, and growth temperature. Additionally, cell lysis, sonication and salt precipitation conditions were optimized. Peptide expression was confirmed by Western blot and Coomassie techniques. The peptide was purified by ammonium sulfate, ion-exchange chromatography and nickel column techniques. The peptide was successfully delivered to human VSM and HEK-293 cells, confirmed by microscopic visualization and western blot techniques, which will be utilized to assess activity of endogenous α2C-ARs. Additionally, these studies have broader implications for understanding the role of actin in protein translocation and cell surface delivery.American Physiology Society - Undergraduate Research FellowshipOhio State University - Undergraduate Research ScholarshipNo embarg

Fracture Mechanics Characterization of an AnisotropicGeomaterial

Argillites are considered worldwide as potential host rock for high level radioactive waste given the low permeability and strong adsorption potential. However, the excavation of the galleries of a repository would produce a disturbed zone around the boundaries rich of new fractures which may enhance the conductivity of the rock along the gallery axis. Several mine-by experiments have been performed in underground rock labs to investi- gate the features of the disturbed zone. In Mont Terri URL (Kanton Jura, Switzerland) the EZ-B experiment was specifically conceived for the measurement of excavation induced fractures around a small chamber. The host rock of the URL is a particularly compact and resistant argillite, known as the Opalinus Clay (OPA) excavated and OPA samples were subjected to fracture mechanics tests at the rock mechanics lab of IGAG-CNR in Torino, Italy. The tests aimed at the understanding aspects of the fracturing process occurring in OPA of Mont Terri, which may be considered a transversely isotropic geomaterial, whose planes of isotropy coincide with the beddin

An Open-Source Code For Fluid Flow Simulations In Unconventional Fractured Reservoirs

In this article, an open-source code for the simulation of fluid flow, including adsorption, transport, and indirect hydromechanical coupling in unconventional fractured reservoirs is described. The code leverages cutting-edge numerical modeling capabilities like automatic differentiation, stochastic fracture modeling, multicontinuum modeling, and discrete fracture models. In the fluid mass balance equation, specific physical mechanisms, unique to organic-rich source rocks, are included, like an adsorption isotherm, a dynamic permeability-correction function, and an Embedded Discrete Fracture Model (EDFM) with fracture-to-well connectivity. The code is validated against an industrial simulator and applied for a study of the performance of the Barnett shale reservoir, where adsorption, gas slippage, diffusion, indirect hydromechanical coupling, and propped fractures are considered. It is the first open-source code available to facilitate the modeling and production optimization of fractured shale-gas reservoirs. The modular design also facilitates rapid prototyping and demonstration of new models. This article also contains a quantitative analysis of the accuracy and limitations of EDFM for gas production simulation in unconventional fractured reservoirs

Hydrocomplexity, and spatial and time scales as drivers of monitoring and management approaches for a karst coastal aquifer (Salento, southern Italy)

Hydrogeological scenario, Salento aquifer, monitoring wells

Rainwater Harvesting for Agricultural Irrigation: An Analysis of Global Research

Within a context of scarce water resources for agriculture, rainwater harvesting constitutes a promising alternative that has been studied by different disciplines in recent years. This article analyses the dynamics of global research on rainwater harvesting for agricultural irrigation over the last two decades. To do this, qualitative systematic analysis and quantitative bibliometric analysis have been carried out. The results reveal that this line of research is becoming increasingly important within research on irrigation. Environmental sciences and agricultural and biological sciences are the most relevant subject areas. Agricultural Water Management, Physics and Chemistry of the Earth, and Irrigation and Drainage are the journals that have published the most articles on the subject. India, China, the United States (USA), South Africa, and the Netherlands are the countries that lead this line of research. Although significant progress has been made in this subject area, it is necessary to increase the number of studies on the capacity of rainwater harvesting systems to cover irrigation needs in different farming contexts, the factors that determine their adoption by farmers, the economic and financial feasibility of their implementation, and their contribution to mitigating global climate change

Reader Time Investment as a Partial Impact Measure of Online Extension Content

Information published online can help many vineyard workers better understand concepts that relate to job performance. The eXtension Grape Community of Practice (GCoP) created numerous articles and other content to extend information to their community of interest. Assessing impact of these interactions is difficult; however, using economic models such as opportunity cost could help assign monetary value to each page view. Although it presents an incomplete picture of the impact of the article, deriving the opportunity cost allows authors of online articles to assess how readers value their time and the investment they are willing to make to read online information

Sustainable Irrigation in Agriculture: An Analysis of Global Research

Irrigated agriculture plays a fundamental role as a supplier of food and raw materials. However, it is also the world’s largest water user. In recent years, there has been an increase in the number of studies analyzing agricultural irrigation from the perspective of sustainability with a focus on its environmental, economic, and social impacts. This study seeks to analyze the dynamics of global research in sustainable irrigation in agriculture between 1999 and 2018, including the main agents promoting it and the topics that have received the most attention. To do this, a review and a bibliometric analysis were carried out on a sample of 713 articles. The results show that sustainability is a line of study that is becoming increasingly more prominent within research in irrigation. The study also reveals the existence of substantial differences and preferred topics in the research undertaken by different countries. The priority issues addressed in the research were climatic change, environmental impact, and natural resources conservation; unconventional water resources; irrigation technology and innovation; and water use efficiency. Finally, the findings indicate a series of areas related to sustainable irrigation in agriculture in which research should be promoted

The recent floods in the Asso Torrent basin (Apulia, Italy): An investigation to improve the stormwater management

Stormwater management is of concern to public institutions and academies. In the Asso Torrent endorheic basin (Salento peninsula, Southern Italy), a network of natural and artificial channels crossing urban and rural areas and flowing towards six swallow karst holes, several floods occurred in the last six years, after the end of extensive hydraulic works. In this paper, the results of an observational study on the meteorological and hydrological factors concurring to these floods are reported. It was inferred that soil saturation, cumulative precipitation anticipating the events, and clogging of the swallow holes are of relevance and must be considered in the definition of new criteria in decision-making procedure. The adoption of both innovative modeling techniques and real-time control should be an efficient solution to properly regulate the flow control devices before and during the precipitation events. With a view to providing solutions for a sustainable management of the water resources, easy-to-implement measures are suggested, such as the selection of flood-tolerant crops and construction of harvesting systems for alternative water uses

La caduta dello stato eroico. Il conflitto nella Scienza Nuova di Giambattista Vico

The aim of this essay is to analyze the figure of conflict in Giambattista Vico's Scienza Nuova, highlighting its epistemological function and its historical-political cogency. This conflict presents itself both as an epistemological means through which reason understands the process of community establishment, and as the effective force for the development of nations. In fact, the changing of times outlined by Vico takes the form of an asymmetrical conflict which proves to be socially significant to the extent that it allows the recomposition of the social body within an order marked by the recognition, albeit precarious, of equality