The α\u3csub\u3e1D\u3c/sub\u3e-Adrenergic Receptor Is Expressed Intracellularly and Coupled to Increases in Intracellular Calcium and Reactive Oxygen Species in Human Aortic Smooth Muscle Cells

Background: The cellular localization of the α1D-adrenergic receptor (α1D-AR) is controversial. Studies in heterologous cell systems have shown that this receptor is expressed in intracellular compartments. Other studies show that dimerization with other ARs promotes the cell surface expression of the α1D-AR. To assess the cellular localization in vascular smooth muscle cells, we developed an adenoviral vector for the efficient expression of a GFP labeled α1D-AR. We also measured cellular localization with immunocytochemistry. Intracellular calcium levels, measurement of reactive oxygen species and contraction of the rat aorta were used as measures of functional activity. Results: The adenovirally expressed α1D-AR was expressed in intracellular compartments in human aortic smooth muscle cells. The intracellular localization of the α1D-AR was also demonstrated with immunocytochemistry using an α1D-AR specific antibody. RT-PCR analysis detected mRNA transcripts corresponding to the α1A-α1B- and α1D-ARs in these aortic smooth muscle cells. Therefore, the presence of the other α1-ARs, and the potential for dimerization with these receptors, does not alter the intracellular expression of the α1D-AR. Despite the predominant intracellular localization in vascular smooth muscle cells, the α1D-AR remained signaling competent and mediated the phenylephrine-induced increases in intracellular calcium. The α1D-AR also was coupled to the generation of reactive oxygen species in smooth muscle cells. There is evidence from heterologous systems that the α1D-AR heterodimerizes with the β2-AR and that desensitization of the β2-AR results in α1D-AR desensitization. In the rat aorta, desensitization of the β2-AR had no effect on contractile responses mediated by the α1D-AR. Conclusion: Our results suggest that the dimerization of the α1D-AR with other ARs does not alter the cellular expression or functional response characteristics of the α1D-AR

The α1D-adrenergic receptor is expressed intracellularly and coupled to increases in intracellular calcium and reactive oxygen species in human aortic smooth muscle cells

Background: The cellular localization of the α1D-adrenergic receptor (α1D-AR) is controversial. Studies in heterologous cell systems have shown that this receptor is expressed in intracellular compartments. Other studies show that dimerization with other ARs promotes the cell surface expression of the α1D-AR. To assess the cellular localization in vascular smooth muscle cells, we developed an adenoviral vector for the efficient expression of a GFP labeled α1D-AR. We also measured cellular localization with immunocytochemistry. Intracellular calcium levels, measurement of reactive oxygen species and contraction of the rat aorta were used as measures of functional activity. Results: The adenovirally expressed α1D-AR was expressed in intracellular compartments in human aortic smooth muscle cells. The intracellular localization of the α1D-AR was also demonstrated with immunocytochemistry using an α1D-AR specific antibody. RT-PCR analysis detected mRNA transcripts corresponding to the α1A-α1B- and α1D-ARs in these aortic smooth muscle cells. Therefore, the presence of the other α1-ARs, and the potential for dimerization with these receptors, does not alter the intracellular expression of the α1D-AR. Despite the predominant intracellular localization in vascular smooth muscle cells, the α1D-AR remained signaling competent and mediated the phenylephrine-induced increases in intracellular calcium. The α1D-AR also was coupled to the generation of reactive oxygen species in smooth muscle cells. There is evidence from heterologous systems that the α1D-AR heterodimerizes with the β2-AR and that desensitization of the β2-AR results in α1D-AR desensitization. In the rat aorta, desensitization of the β2-AR had no effect on contractile responses mediated by the α1D-AR. Conclusion: Our results suggest that the dimerization of the α1D-AR with other ARs does not alter the cellular expression or functional response characteristics of the α1D-AR

Parent-Reported Clinical Utility of Pediatric Genomic Sequencing.

Background and objectivesGenomic sequencing (GS) is increasingly used for diagnostic evaluation, yet follow-up care is not well understood. We assessed clinicians' recommendations after GS, parent-reported follow-up, and actions parents initiated in response to learning their child's GS results.MethodsWe surveyed parents of children who received GS through the Clinical Sequencing Evidence Generating Research consortium ∼5 to 7 months after return of results. We compared the proportion of parents who reported discussing their child's result with a clinician, clinicians' recommendations, and parents' follow-up actions by GS result type using χ2 tests.ResultsA total of 1188 respondents completed survey measures on recommended medical actions (n = 1187) and/or parent-initiated actions (n = 913). Most parents who completed recommended medical actions questions (n = 833, 70.3%) reported having discussed their child's GS results with clinicians. Clinicians made recommendations to change current care for patients with positive GS results (n = 79, 39.1%) more frequently than for those with inconclusive (n = 31, 12.4%) or negative results (n = 44, 11.9%; P < .001). Many parents discussed (n = 152 completed, n = 135 planned) implications of GS results for future pregnancies with a clinician. Aside from clinical recommendations, 13.0% (n = 119) of parents initiated changes to their child's health or lifestyle.ConclusionsIn diverse pediatric clinical contexts, GS results can lead to recommendations for follow-up care, but they likely do not prompt large increases in the quantity of care received

Beltline: A History of the Atlanta Beltline and its Associated Historic Resources

Prepared by the Spring 2006 Preservation Planning Class. This project, developed in a collaborative effort by the Atlanta Urban Design commission and GSU students, was designed to highlight the history of the proposed redevelopment nodes along the Beltline, which is an outer band railroad line encircling the city. The purpose was to identify the significance of the resources therein and their links to Atlanta’s history, providing assistance in the overall development process of the Beltline project.https://scholarworks.gsu.edu/history_heritagepreservation/1003/thumbnail.jp

Competency-based evaluation tools for integrative medicine training in family medicine residency: a pilot study

BACKGROUND: As more integrative medicine educational content is integrated into conventional family medicine teaching, the need for effective evaluation strategies grows. Through the Integrative Family Medicine program, a six site pilot program of a four year residency training model combining integrative medicine and family medicine training, we have developed and tested a set of competency-based evaluation tools to assess residents' skills in integrative medicine history-taking and treatment planning. This paper presents the results from the implementation of direct observation and treatment plan evaluation tools, as well as the results of two Objective Structured Clinical Examinations (OSCEs) developed for the program. METHODS: The direct observation (DO) and treatment plan (TP) evaluation tools developed for the IFM program were implemented by faculty at each of the six sites during the PGY-4 year (n = 11 on DO and n = 8 on TP). The OSCE I was implemented first in 2005 (n = 6), revised and then implemented with a second class of IFM participants in 2006 (n = 7). OSCE II was implemented in fall 2005 with only one class of IFM participants (n = 6). Data from the initial implementation of these tools are described using descriptive statistics. RESULTS: Results from the implementation of these tools at the IFM sites suggest that we need more emphasis in our curriculum on incorporating spirituality into history-taking and treatment planning, and more training for IFM residents on effective assessment of readiness for change and strategies for delivering integrative medicine treatment recommendations. Focusing our OSCE assessment more narrowly on integrative medicine history-taking skills was much more effective in delineating strengths and weaknesses in our residents' performance than using the OSCE for both integrative and more basic communication competencies. CONCLUSION: As these tools are refined further they will be of value both in improving our teaching in the IFM program and as competency-based evaluation resources for the expanding number of family medicine residency programs incorporating integrative medicine into their curriculum. The next stages of work on these instruments will involve establishing inter-rater reliability and defining more clearly the specific behaviors which we believe establish competency in the integrative medicine skills defined for the program

Design, fabrication, and performance of infrared and visible vertical cavity surface emitting lasers”,

Abstract-This paper discusses the issues involving the design and fabrication of vertical-cavity surface-emitting lasers (VCSEL's). A review of the basic experimental structures is given, with emphasis on recent developments in distributed Bragg reflectors, gain media, as well as current and optical confinement techniques. The paper describes present VCSEL performance, in particular, those involving selective oxidation and visible wavelength operation

Effect of follower load on motion and stiffness of the human thoracic spine with intact rib cage

Researchers have reported on the importance of the rib cage in maintaining mechanical stability in the thoracic spine and on the validity of a compressive follower preload. However, dynamic mechanical testing using both the rib cage and follower load has never been studied. An in vitro biomechanical study of human cadaveric thoracic specimens with rib cage intact in lateral bending, flexion/extension, and axial rotation under varying compressive follower preloads was performed. The objective was to characterize the motion and stiffness of the thoracic spine with intact rib cage and follower preload. The hypotheses tested for all modes of bending were (i) range of motion, elastic zone, and neutral zone will be reduced with a follower load, and (ii) neutral and elastic zone stiffness will be increased with a follower load. Eight human cadaveric thoracic spine specimen (T1–T12) with intact rib cage were subjected to 5 Nm pure moments in lateral bending, flexion/extension, and axial rotation under follower loads of 0–400 N. Range of motion, elastic and neutral zones, and elastic and neutral zone stiffness values were calculated for functional spinal units and segments within the entire thoracic section. Combined segmental range of motion decreased by an average of 34% with follower load for every mode. Application of a follower load with intact rib cage impacts the motion and stiffness of the human cadaveric thoracic spine. Researchers should consider including both aspects to better represent the physiologic implications of human motion and improve clinically relevant biomechanical thoracic spine testing

Effects of follower load and rib cage on intervertebral disc pressure and sagittal plane curvature in static tests of cadaveric thoracic spines

The clinical relevance of mechanical testing studies of cadaveric human thoracic spines could be enhanced by using follower preload techniques, by including the intact rib cage, and by measuring thoracic intervertebral disc pressures, but studies to date have not incorporated all of these components simultaneously. Thus, this study aimed to implement a follower preload in the thoracic spine with intact rib cage, and examine the effects of follower load, rib cage stiffening and rib cage removal on intervertebral disc pressures and sagittal plane curvatures in unconstrained static conditions. Intervertebral disc pressures increased linearly with follower load magnitude. The effect of the rib cage on disc pressures in static conditions remains unclear because testing order likely confounded the results. Disc pressures compared well with previous reports in vitro, and comparison with in vivo values suggests the use of a follower load of about 400 N to approximate loading in upright standing. Follower load had no effect on sagittal plane spine curvature overall, suggesting successful application of the technique, although increased flexion in the upper spine and reduced flexion in the lower spine suggest that the follower load path was not optimized. Rib cage stiffening and removal both increased overall spine flexion slightly, although with differing effects at specific spinal locations. Overall, the approaches demonstrated here will support the use of follower preloads, intact rib cage, and disc pressure measurements to enhance the clinical relevance of future studies of the thoracic spine