Hormonal Regulation of Capillary Fenestrae in the Rat Adrenal Cortex: Quantitative Studies using Objective Lens Staging Scanning Electron Microscopy

High magnification studies of the fenestrated capillary endothelium in the zona fasciculata (ZF) of rat adrenal glands were performed using the objective lens stage of an analytical scanning electron microscope (SEM) equipped with a lanthanum hexaboride emitter (LaB6). Resolution of surface substructure of the luminal membrane obtained with specimens decorated with gold/palladium (Au/Pd) was compared with that observed in others sputter coated with tantalum (Ta). High magnification (50,000x) of the fenestrated endothelium demonstrates that tantalum coating of the cryofractured adrenals improves the substructural detail compared to that seen in Au/Pd decorated specimens. The procedures used in specimen preparation, metal deposition and secondary electron imaging (SEI) are described. Quality imaging achieved using the objective lens stage is a result of the elimination of the SE-III component derived from backscattered electrons. Rat adrenals exhibited uniformly patent capillaries. High magnification micrographs of capillary walls were randomly recorded in two morphometric studies of the fenestral content of capillaries in the rat adrenal cortex. Adrenocorticotropic hormone (ACTH), when administered to rats following dexamethasone (DEX) treatment, significantly reduced the fenestrae/μm2 of endothelial surface and increased the mean size of fenestrae. After hypophysectomy, the number of fenestrae/μm2 declined over 48 h; within 2 h after ACTH was given to rats hypophysectomized 48 hours earlier, the fenestrae/μm2 had increased two-fold. These studies indicate that ACTH plays an important role in modulating fenestral content of the capillary endothelium in the adrenal cortex

Correlative Light, Transmission, and High Resolution (SE-I) Scanning Electron Microscopy Studies of Rhesus Adrenocortical Vascular Morphology

A detailed correlative morphologic description using light microscopy (LM), transmission electron microscopy (TEM) and high resolution SE-I scanning electron microscopy (SEM) was conducted on the capillary endothelium of the zona-fasciculata (Z-F) in juvenile male rhesus monkeys. The glucocorticoid synthesis and release phenomena, associated with stress stimulated release of adrenocorticotropic hormone (ACTH) via the hypothalamic-pituitary axis, intimately involves capillaries of the Z-F. A comprehensive study of all the ultrastructural features implicated in the transendothelial uptake of steroidogenic precursors and release of glucocorticoids in perfused rhesus adrenals has not previously been made. This report presents correlative images of transendothelial openings that include previously described single diaphragmed fenestrae and plasmalemma vesicles, and double diaphragmed transendothelial channels. New observations of endothelial cell pockets, tight junctional complexes and membrane filled ghost sacs were recorded from perfused rhesus adrenal. Membranous ghosts associated with adrenocortical endothelium were reported in a previous TEM study of perfused rat, however the potential argument existed that ghosts were artifactual. Their role as steroid hormone releasing structures remains an open question, yet their structural characteristics appear justified based on imaging of identical profiles observed in perfused rhesus adrenocortical specimens. These structural features are considered for the potential of gating and sorting of metabolites, and release of glucocorticoids in response to ACTH stimulated stress events

Correlated Microscopic Observations of Arterial Responses to Intravascular Stenting

Percutaneous catheter implantation of intravascular stent prostheses has emerged as a novel clinical adjunct to balloon angioplasty in the treatment of obstructive atherosclerotic vascular disease. We have examined the cellular and subcellular responses to stenting in the coronary arteries of the dog and pig (both normal and atherosclerotic), and in the iliac arteries and aorta of the atherosclerotic rabbit, using scanning electron, transmission electron, and light microscopies. Stenting in these models resulted in a thrombotic reaction ranging from mild to severe, depending on species and antithrombotic therapy. Subsequent organization of thrombotic material with hyperplasia of smooth muscle and inflammatory cells, luminal recovering with endothelial or pseudoendothelial cells, and atrophy of the tunica media led to incorporation of the prosthesis into the arterial wall. Endothelial or pseudoendothelial cells were observed adherent to the prosthesis as early as one day after placement, and regeneration of a confluent periluminal cell layer occurred within 2 to 4 weeks. Persistent ultrastructural abnormalities of the periluminal cell layer were seen as late as 2 years after stenting, but the intimal hyperplastic response appeared limited

Energy-Filtering Transmission Electron Microscopy as a Tool for Structural and Compositional Analysis of Isolated Ferritin Particles

Structural and compositional analysis of isolated horse-spleen ferritin particles was performed by energy filtering transmission electron microscopy (EFTEM). Ferritin particles were collected in ultrathin (2 nm thick) chromium films and analyzed without any additional stain by electron energy-loss spectroscopy (EELS) for iron and carbon and by electron-spectroscopic imaging (ESI) for carbon. The ultrastructure of the proteinaceous shell of the ferritin particle, as obtained by the carbon net-intensity electron spectroscopical and carbon concentration-distribution images, was qualitatively compared to the structure as acquired by a negative-staining procedure. Quantitative analysis of the number of carbon atoms in the ferritin-shell proteins was carried out through an ESI-acquisition protocol and processing procedure with calibrated attenuation filters in the optical path to the TV camera. This procedure included images acquired with calibrated attenuation filters for the compensation of shading and the non-linear performance of the TV camera used in the analytical part of the procedure. A new ESI-Spectra program is proposed that allows element-related spectra to be generated at any place and with any frame size in a contrast-sensitive or other type of image present on the computer monitor screen

Neurodegenerative Properties of Chronic Pain: Cognitive Decline in Patients with Chronic Pancreatitis

Chronic pain has been associated with impaired cognitive function. We examined cognitive performance in patients with severe chronic pancreatitis pain. We explored the following factors for their contribution to observed cognitive deficits: pain duration, comorbidity (depression, sleep disturbance), use of opioids, and premorbid alcohol abuse. The cognitive profiles of 16 patients with severe pain due to chronic pancreatitis were determined using an extensive neuropsychological test battery. Data from three cognitive domains (psychomotor performance, memory, executive functions) were compared to data from healthy controls matched for age, gender and education. Multivariate multilevel analysis of the data showed decreased test scores in patients with chronic pancreatitis pain in different cognitive domains. Psychomotor performance and executive functions showed the most prominent decline. Interestingly, pain duration appeared to be the strongest predictor for observed cognitive decline. Depressive symptoms, sleep disturbance, opioid use and history of alcohol abuse provided additional explanations for the observed cognitive decline in some of the tests, but to a lesser extent than pain duration. The negative effect of pain duration on cognitive performance is compatible with the theory of neurodegenerative properties of chronic pain. Therefore, early and effective therapeutic interventions might reduce or prevent decline in cognitive performance, thereby improving outcomes and quality of life in these patients

Examining whether and how instructional coordination occurs within introductory undergraduate STEM courses

Instructors’ interactions can foster knowledge sharing around teaching and the use of research-based instructional strategies (RBIS). Coordinated teaching presents an impetus for instructors’ interactions and creates opportunities for instructional improvement but also potentially limits an instructor’s autonomy. In this study, we sought to characterize the extent of coordination present in introductory undergraduate courses and to understand how departments and instructors implement and experience course coordination. We examined survey data from 3,641 chemistry, mathematics, and physics instructors at three institution types and conducted follow-up interviews with a subset of 24 survey respondents to determine what types of coordination existed, what factors led to coordination, how coordination constrained instruction, and how instructors maintained autonomy within coordinated contexts. We classified three approaches to coordination at both the overall course and course component levels: independent (i.e., not coordinated), collaborative (decision-making by instructor and others), controlled (decision-making by others, not instructor). Two course components, content coverage and textbooks, were highly coordinated. These curricular components were often decided through formal or informal committees, but these decisions were seldom revisited. This limited the ability for instructors to participate in the decision-making process, the level of interactions between instructors, and the pedagogical growth that could have occurred through these conversations. Decision-making around the other two course components, instructional methods and exams, was more likely to be independently determined by the instructors, who valued this autonomy. Participants in the study identified various ways in which collaborative coordination of courses can promote but also inhibit pedagogical growth. Our findings indicate that the benefits of collaborative course coordination can be realized when departments develop coordinated approaches that value each instructor’s autonomy, incorporate shared and ongoing decision-making, and facilitate collaborative interactions and knowledge sharing among instructors

Examining whether and how instructional coordination occurs within introductory undergraduate STEM courses

Instructors’ interactions can foster knowledge sharing around teaching and the use of research-based instructional strategies (RBIS). Coordinated teaching presents an impetus for instructors’ interactions and creates opportunities for instructional improvement but also potentially limits an instructor’s autonomy. In this study, we sought to characterize the extent of coordination present in introductory undergraduate courses and to understand how departments and instructors implement and experience course coordination. We examined survey data from 3,641 chemistry, mathematics, and physics instructors at three institution types and conducted follow-up interviews with a subset of 24 survey respondents to determine what types of coordination existed, what factors led to coordination, how coordination constrained instruction, and how instructors maintained autonomy within coordinated contexts. We classified three approaches to coordination at both the overall course and course component levels: independent (i.e., not coordinated), collaborative (decisionmaking by instructor and others), controlled (decision-making by others, not instructor). Two course components, content coverage and textbooks, were highly coordinated. These curricular components were often decided through formal or informal committees, but these decisions were seldom revisited. This limited the ability for instructors to participate in the decision-making process, the level of interactions between instructors, and the pedagogical growth that could have occurred through these conversations. Decision-making around the other two course components, instructional methods and exams, was more likely to be independently determined by the instructors, who valued this autonomy. Participants in the study identified various ways in which collaborative coordination of courses can promote but also inhibit pedagogical growth. Our findings indicate that the benefits of collaborative course coordination can be realized when departments develop coordinated approaches that value each instructor’s autonomy, incorporate shared and ongoing decision-making, and facilitate collaborative interactions and knowledge sharing among instructors

The translation, validity and reliability of the German version of the Fremantle Back Awareness Questionnaire

Background: The Fremantle Back Awareness Questionnaire (FreBAQ) claims to assess disrupted self-perception of the back. The aim of this study was to develop a German version of the Fre-BAQ (FreBAQ-G) and assess its test-retest reliability, its known-groups validity and its convergent validity with another purported measure of back perception. Methods: The FreBaQ-G was translated following international guidelines for the transcultural adaptation of questionnaires. Thirty-five patients with non-specific CLBP and 48 healthy participants were recruited. Assessor one administered the FreBAQ-G to each patient with CLBP on two separate days to quantify intra-observer reliability. Assessor two administered the FreBaQ-G to each patient on day 1. The scores were compared to those obtained by assessor one on day 1 to assess inter-observer reliability. Known-groups validity was quantified by comparing the FreBAQ-G score between patients and healthy controls. To assess convergent validity, patient\u27s FreBAQ-G scores were correlated to their two-point discrimination (TPD) scores. Results: Intra- and Inter-observer reliability were both moderate with ICC3.1 = 0.88 (95%CI: 0.77 to 0.94) and 0.89 (95%CI: 0.79 to 0.94), respectively. Intra- and inter-observer limits of agreement (LoA) were 6.2 (95%CI: 5.0±8.1) and 6.0 (4.8±7.8), respectively. The adjusted mean difference between patients and controls was 5.4 (95%CI: 3.0 to 7.8, p\u3c0.01). Patient\u27s FreBAQ-G scores were not associated with TPD thresholds (Pearson\u27s r = -0.05, p = 0.79). Conclusions: The FreBAQ-G demonstrated a degree of reliability and known-groups validity. Interpretation of patient level data should be performed with caution because the LoA were substantial. It did not demonstrate convergent validity against TPD. Floor effects of some items of the FreBAQ-G may have influenced the validity and reliability results. The clinimetric properties of the FreBAQ-G require further investigation as a simple measure of disrupted self-perception of the back before firm recommendations on its use can be made

Mapping Brain Response to Pain in Fibromyalgia Patients Using Temporal Analysis of fMRI

Background: Nociceptive stimuli may evoke brain responses longer than the stimulus duration often partially detected by conventional neuroimaging. Fibromyalgia patients typically complain of severe pain from gentle stimuli. We aimed to characterize brain response to painful pressure in fibromyalgia patients by generating activation maps adjusted for the duration of brain responses. Methodology/Principal Findings: Twenty-seven women (mean age: 47.8 years) were assessed with fMRI. The sample included nine fibromyalgia patients and nine healthy subjects who received 4 kg/cm2 of pressure on the thumb. Nine additional control subjects received 6.8 kg/cm2 to match the patients for the severity of perceived pain. Independent Component Analysis characterized the temporal dynamics of the actual brain response to pressure. Statistical parametric maps were estimated using the obtained time courses. Brain response to pressure (18 seconds) consistently exceeded the stimulus application (9 seconds) in somatosensory regions in all groups. fMRI maps following such temporal dynamics showed a complete pain network response (sensory-motor cortices, operculo-insula, cingulate cortex, and basal ganglia) to 4 kg/cm2 of pressure in fibromyalgia patients. In healthy subjects, response to this low intensity pressure involved mainly somatosensory cortices. When matched for perceived pain (6.8 kg/cm2), control subjects showed also comprehensive activation of pain-related regions, but fibromyalgia patients showed significantly larger activation in the anterior insula-basal ganglia complex and the cingulate cortex. Conclusions/Significance: The results suggest that data-driven fMRI assessments may complement conventional neuroimaging for characterizing pain responses and that enhancement of brain activation in fibromyalgia patients may be particularly relevant in emotion-related regions