Evaluating techniques in tissue clarification using CLARITY imaging and investigating where sodium is sensed in the body

OBJECTIVE: Previous studies have shown the significant contribution of sympathoinhibition in response to sodium loading to prevent increases in mean arterial blood pressure in salt resistant phenotypes. It has also been shown that brain Gαi2 protein gated signal transduction plays a major role in this pathway, however, the specific mechanisms through which this pathway is activated remain less well understood. The purpose of this study was to elucidate the relative contribution of increased sodium in either the plasma or the cerebrospinal fluid (CSF) to the regulation of mean arterial pressure and natriuresis. Additionally we explored the potential for using the novel CLARITY Imaging technique to identify the relative activity of neurons in areas of the brain thought to play a major role in body fluid homeostasis in response to salt. METHODS: Rats that were pre-treated with either scrambled or Gαi2 oligodeoxynucleotides (ODN), to selectively down regulate brain Gαi2 proteins, were challenged either peripherally or centrally with sodium. Upon sodium loading physiological parameters were measured for two hours after which the animal's brains were recovered for immunohistochemical (IHC) analysis of the paraventricular nucleus, a known regulatory center for body fluid homeostasis and blood pressure regulation. Additionally we adapted a version of the published CLARITY Imaging protocols for optically clearing tissue through application of electrophoretic tissue clearing (ETC) to a larger rat model. RESULTS: In scrambled ODN pre-treated rats we observed a temporary increase in MAP in response to both the peripheral and central sodium challenge. In the Gαi2 ODN pre-treated animals we saw some form of attenuation to this response in both studies, however, where in the peripheral challenge there was an increase in the amount of time that it took the rats to return to normotension with no alteration in natriuresis, in the central challenge there was a large attenuation in natriuresis with no differences in the time to return to baseline MAP. Our IHC analysis also showed a decrease in neuronal activation of paraventricular medial parvocellular neurons in Gαi2 pre-treated rats that were challenged peripherally vs their SCR pre-treated counterparts. No such difference was observed in either of the pre-treatment groups from the central sodium challenge study. In the CLARITY study we found that it is possible to adapt the method for optically clearing tissue to the larger model, however, we encountered several issues related to tissue swelling and peripheral tissue damage. CONCLUSION: Based on our current results it seems evident that there are at least two different mechanisms that activate the cardiovascular regulatory control centers in the brain that prevent long term increases in mean arterial pressure in response to increased salt. It also appears that these two different mechanisms are triggered either by increases in plasma or CSF salinity, though which of these two mechanisms may be directly responsible for the development of salt sensitive hypertension requires further investigation. While we had some success at optically clearing larger tissue volumes through ETC, problems we encountered with maintaining tissue integrity for investigations of intact neural networks prevented us from applying this technique, in its current form, to our investigation of salt sensitive hypertension

Electronic Construction Coordination with AutoCAD

A key component to the construction process is drawing coordination. Due to the fast paced nature of today’s construction, many design firms utilize other consultants to aid in completion of the documents within the required design time frame. Coordination between consultants is often neglected as the design is completed. Additionally, traditional coordination methods utilize hard copies and light tables for coordination of construction drawings. Therefore, it is desirable to create an improvement plan that utilizes electronic coordination for the construction process. Modifying the coordination process to a soft format that utilizes AutoCAD design software would improve quality and accuracy. The improvement plan also included changes to the bid process and job buyout. Shop drawing development was required to be completed utilizing zone and layer control. Drawing guidelines were developed to simplify compiling of multiple files into a single drawing. A key aspect of this project was document management. Files were produced using specified layer control, labeling requirements, and drawing standards. A third party was hired to manage an FTP site for storage and movement of files. This system ensured the current files were available for use and the older files were archived. The construction coordination process requires three stages, drawing coordination, field coordination, and as-built development. The majority of coordination is completed during the drawing coordination stage. Field coordination is utilized to correct for errors and problems created by overlapping tolerances and installation errors. The work in place is then documented to insure the drawings are updated to provide accurate as-built drawings for the client. The program provided improvements to the existing system and achieved a basic level of soft format coordination. This system improved speed, lower costs, and allowed individual sketches of specific items to be produced for review. The use of layer control allowed for plots to be produced showing only the layers critical to the specified application. This uncluttered the view and made a more useful drawing. The information learned from implementing this process has identified areas that should be improved prior to repeating the process. Additional time should be devoted to training participants in the use of the available resources, labeling conventions, naming conventions, and drawing standards. Other areas that created problems were inconsistency in the drawing packages and third-party plug-ins used by subcontractors. Further investigation identified many of the plug-ins were required to support their shop manufacturing processes and could not be eliminated. Further work would be to use the information gained during this project for improvement of the implementation of a Building Information Management (BIM) system. The architectural design packages are currently set for developing drawings in 3D. The BIM system adds a database of critical component information which is packaged with the drawing. The mechanical and electrical trades have not adapted the new formats for design at this time. The mechanical and electrical software designers are still in work on development of the BIM design packages. Once this step is complete, the program will contain all critical parts to being a fully operation system

The Utilization Of Local History In Teaching American Religious History: A Gilded Age And Progressive Era North Dakota Case Study

Teachers of college-level courses on American religious history generally leave out the importance of local and regional histories when telling the story of religion in America. The study of local history provides a fertile ground for understanding broad national trends in a local context. This dissertation focuses upon a little-studied religious body in North Dakota to see how Anglo settlers on the prairie viewed their religious experience in a Gilded Age and Progressive Era context. By emphasizing the records of the North Dakota Baptist State Convention at the North Dakota State Archives in Bismarck and the First Baptist Church of Grand Forks at the Orin G. Libby Department of Special Collections, it quickly became evident that the North Dakota Baptists fell solidly in line with the overarching English-speaking Protestant establishment that supported such efforts and ideas as evangelism, reform, nativism, and the Great War as the pathway to a truly Christianized America. As a Doctor of Arts project, this research was integrated into a course on Religion in American Politics and Culture. The results of this case study in terms of student learning and assessment make up the conclusion of this project

Explorations in engagement for humans and robots

This paper explores the concept of engagement, the process by which individuals in an interaction start, maintain and end their perceived connection to one another. The paper reports on one aspect of engagement among human interactors--the effect of tracking faces during an interaction. It also describes the architecture of a robot that can participate in conversational, collaborative interactions with engagement gestures. Finally, the paper reports on findings of experiments with human participants who interacted with a robot when it either performed or did not perform engagement gestures. Results of the human-robot studies indicate that people become engaged with robots: they direct their attention to the robot more often in interactions where engagement gestures are present, and they find interactions more appropriate when engagement gestures are present than when they are not.Comment: 31 pages, 5 figures, 3 table

Multi-Purpose ESS/ITS Data Collection Sites

This document presents the results of a state-of-practice survey of transportation agencies that are installing intelligent transportation sensors (ITS) and other devices along with their environmental sensing stations (ESS) also referred to as roadway weather information system (RWIS) assets