In vivo approaches to investigate ANCA-associated vasculitis: lessons and limitations

Anti-neutrophil cytoplasm autoantibody (ANCA)-associated diseases are autoimmune conditions characterized by necrotizing inflammation of small blood vessels. The immunogenesis and etiology of these conditions are unknown, but our knowledge of the immunopathogenesis has increased considerably in recent years. In this review, we discuss the animal models currently used to investigate the mechanisms of vascular injury and to test novel therapies. We outline their advantages and limitations and propose potential directions for future research

Thrombin-mediated proteoglycan synthesis utilizes both protein-tyrosine kinase and serine/threonine kinase receptor transactivation in vascular smooth muscle cells

Background: GPCR transactivation of PTKRs and TGF-αRs mediates proteoglycan synthesis in human VSMC. Results: Transactivation of TGF-αRs is integrin-dependent, and inhibition of both transactivation pathways blocks proteoglycan synthesis. Conclusion: GPCR utilize transactivation pathways and not classical signaling in proteoglycan synthesis. Significance: GPCR transactivation of receptor kinase pathways may be broader and more significant than previously recognized

A novel 4-dimensional live-cell imaging system to study leukocyte-endothelial dynamics in ANCA-associated vasculitis

This work was supported by the Lauren Currie Twilight Foundation, NHS Grampian Renal Endowment Fund and Aberdeen University Development Fund.Peer reviewedPostprintPostprin

Complex interplay between intrinsic and extrinsic drivers of long-term survival trends in southern elephant seals

BACKGROUND: Determining the relative contribution of intrinsic and extrinsic factors to fluctuations in population size, trends and demographic composition is analytically complex. It is often only possible to examine the combined effects of these factors through measurements made over long periods, spanning an array of population densities or levels of food availability. Using age-structured mark-recapture models and datasets spanning five decades (1950–1999), and two periods of differing relative population density, we estimated age-specific probabilities of survival and examined the combined effects of population density and environmental conditions on juvenile survival of southern elephant seals at Macquarie Island. RESULTS: First-year survival decreased with density during the period of highest population size, and survival increased during years when the Southern Oscillation Index (SOI) anomaly (deviation from a 50-year mean) during the mother's previous foraging trip to sea was positive (i.e., El Niño). However, when environmental stochasticity and density were considered together, the effect of density on first-year survival effectively disappeared. Ignoring density effects also leads to models placing too much emphasis on the environmental conditions prevailing during the naïve pup's first year at sea. CONCLUSION: Our analyses revealed that both the state of the environment and population density combine to modify juvenile survival, but that the degree to which these processes contributed to the variation observed was interactive and complex. This underlines the importance of evaluating the relative contribution of both the intrinsic and extrinsic factors that regulate animal populations because false conclusions regarding the importance of population regulation may be reached if they are examined in isolation

What's the Matter with Kansas?: The Development and Confirmation of the Evolutionary Attitudes and Literacy Survey (EALS)

This is the publisher's version, also available electronically from http://link.springer.com/article/10.1007%2Fs12052-010-0294-1.The present survey was designed to assess predominant regional belief systems and the roles these beliefs play in science understanding and attitudes, and curricular effectiveness in colleges and universities. To this end, we created a wide variety of theory-driven subscales (lower order factors) reflecting, for example, exposure to evolutionary material, young earth creationist beliefs, moral and social objections, political ideology, endorsement of intelligent design fallacies, knowledge (and distrust) of the scientific enterprise, and attitudes of evolutionary theory’s relevance in several domains (e.g., sciences and humanities). We also included potentially important demographic variables (e.g., rural upbringing, family size). Finally, we assessed openness to experience, a key facet of personality. Hierarchical Structural Equation Modeling (SEM) analysis showed the 16 constructs to have a theoretically meaningful and quantitatively coherent higher order factor structure. In this large Kansas sample of university students, creationist reasoning and conservative orientation (political and religious) were negatively associated with exposure to evolutionary theory, knowledge about it, and positive attitudes toward its relevance. At the same time, exposure to the theory was positively associated with knowledge and positive attitudes. Importantly, though most Kansas-specific demographic variables (e.g., rural origins) were largely unrelated to outcomes of interest in this university-based sample, the personality factor openness to experience appears to be highly relevant for several higher order factors (e.g., exposure, knowledge and relevance, and creationist reasoning). We close with implications for educators and the next steps in survey development

What's the Matter with Kansas?: The Development and Confirmation of the Evolutionary Attitudes and Literacy Survey (EALS)

This is the publisher's version, also available electronically from http://link.springer.com/article/10.1007%2Fs12052-010-0294-1.The present survey was designed to assess predominant regional belief systems and the roles these beliefs play in science understanding and attitudes, and curricular effectiveness in colleges and universities. To this end, we created a wide variety of theory-driven subscales (lower order factors) reflecting, for example, exposure to evolutionary material, young earth creationist beliefs, moral and social objections, political ideology, endorsement of intelligent design fallacies, knowledge (and distrust) of the scientific enterprise, and attitudes of evolutionary theory’s relevance in several domains (e.g., sciences and humanities). We also included potentially important demographic variables (e.g., rural upbringing, family size). Finally, we assessed openness to experience, a key facet of personality. Hierarchical Structural Equation Modeling (SEM) analysis showed the 16 constructs to have a theoretically meaningful and quantitatively coherent higher order factor structure. In this large Kansas sample of university students, creationist reasoning and conservative orientation (political and religious) were negatively associated with exposure to evolutionary theory, knowledge about it, and positive attitudes toward its relevance. At the same time, exposure to the theory was positively associated with knowledge and positive attitudes. Importantly, though most Kansas-specific demographic variables (e.g., rural origins) were largely unrelated to outcomes of interest in this university-based sample, the personality factor openness to experience appears to be highly relevant for several higher order factors (e.g., exposure, knowledge and relevance, and creationist reasoning). We close with implications for educators and the next steps in survey development

Accurate estimates of absolute left ventricular volumes from equilibrium radionuclide angiographic count data using a simple geometric attenuation correction

To simplify and clarify the methods of obtaining attenuation-corrected equilibrium radionuclide angiographic estimates of absolute left ventricular volumes, 27 patients who also had biplane contrast cineangiography were evaluated. Background-corrected left ventricular end-diastolic and end-systolic counts were obtained by semiautomated variable and hand-drawn regions of interest and were normalized to cardiac cycles processed, frame rate and blood sample counts. Blood sample counts were acquired on (d°) and at a distance (d′) from the collimator. A simple geometric attenuation correction was performed to obtain absolute left ventricular volume estimates.Using blood sample counts obtained at d° or d′, the attentuation.corrected radionuclide left ventricular end-diastolic volume estimates using both region of interest selection methods correlated with the cineangiographic end-diastolic volumes (r = 0.95 to 0.96). However, both mean radionuclide semiautomated variable left ventricular end-diastolic volumes (179 ± 100 [± 1 standard deviation] and 185 ± 102 ml, p < 0.001) were smaller than the average cineangiographic end-diastolic volume (217 ± 102 ml), and both mean hand-drawn left ventricular end-diastolic volumes (212 ± 104 and 220 ± 106 ml) did not differ from the average cineangiographic end-diastolic volume. Using the blood sample counts obtained at d° or d′, the attenuation-corrected radionuclide left ventricular end-systolic volume estimates using both region of interest selection methods correlated with the cineangiographic end-systolic volumes (r = 0.96 to 0.98). Also, using blood sample counts at d°, the mean radionuclide semiautomated variable left ventricular end-systolic volume (116 ± 98 ml, p < 0.05) was less than the average cineangiographic end-systolic volume (128 ± 98 ml), and the other radionuclide end-systolic volumes did not differ from the average cineangiographic end-systolic volume.Therefore, it is concluded that: 1) a simple geometric attenuation-correction of radionuclide left ventricular end-diastolic and end-systolic count data provides accurate estimates of biplane cineangiographic end-diastolic and end-systolic volumes; and 2) the hand-drawn region of interest selection method, unlike the semiautomated variable method that underestimates end-diastolic and end-systolic volumes, provides more accurate estimates of biplane cineangiographic left ventricular volumes irrespective of the distance blood sample counts are acquired from the collimator

IL-6 Induced STAT3 Signalling Is Associated with the Proliferation of Human Muscle Satellite Cells Following Acute Muscle Damage

Although the satellite cell (SC) is a key regulator of muscle growth during development and muscle adaptation following exercise, the regulation of human muscle SC function remains largely unexplored. STAT3 signalling mediated via interleukin-6 (IL-6) has recently come to the forefront as a potential regulator of SC proliferation. The early response of the SC population in human muscle to muscle-lengthening contractions (MLC) as mediated by STAT3 has not been studied.Twelve male subjects (21±2 y; 83±12 kg) performed 300 maximal MLC of the quadriceps femoris at 180°•s(-1) over a 55° range of motion with muscle samples (vastus lateralis) and blood samples (antecubital vein) taken prior to exercise (PRE), 1 hour (T1), 3 hours (T3) and 24 hours (T24) post-exercise. Cytoplasmic and nuclear fractions of muscle biopsies were purified and analyzed for total and phosphorylated STAT3 (p-STAT3) by western blot. p-STAT3 was detected in cytoplasmic fractions across the time course peaking at T24 (p<0.01 vs. PRE). Nuclear total and p-STAT3 were not detected at appreciable levels. However, immunohistochemical analysis revealed a progressive increase in the proportion of SCs expressing p-STAT3 with ∼60% of all SCs positive for p-STAT3 at T24 (p<0.001 vs. PRE). Additionally, cMyc, a STAT3 downstream gene, was significantly up-regulated in SCs at T24 versus PRE (p<0.05). Whole muscle mRNA analysis revealed induction of the STAT3 target genes IL-6, SOCS3, cMyc (peaking at T3, p<0.05), IL-6Rα and GP130 (peaking at T24, p<0.05). In addition, Myf5 mRNA was up-regulated at T24 (p<0.05) with no appreciable change in MRF4 mRNA.We demonstrate that IL-6 induction of STAT3 signaling occurred exclusively in the nuclei of SCs in response to MLC. An increase in the number of cMyc+ SCs indicated that human SCs were induced to proliferate under the control of STAT3 signaling

Mucin biosynthesis: purification and characterization of a mucin beta 6N-acetylglucosaminyltransferase.

We have purified, to apparent homogeneity, a mucin beta 6N-acetylglucosaminyltransferase (beta 6GlcNAc transferase) from bovine tracheal epithelium. Golgi membranes were isolated from a 0.25 M sucrose homogenate of epithelial scrapings by discontinuous sucrose gradient centrifugation. The Golgi membranes were solubilized with 1% Triton X-100 in the presence of 1 mM Gal beta 1-3GalNAc alpha benzyl (Bzl) to stabilize the beta 6GlcNAc transferase. The solubilized enzyme was bound to a UDP-hexanolamine-Actigel-ALD Superflow affinity column equilibrated with 1 mM Gal beta 1-3GalNAc alpha Bzl and 5 mM Mn2+. Elution of the enzyme with 0.5 mM UDP-GlcNAc resulted in a 133,800-fold purification with a 1.3% yield and a specific activity of 70 mumol/min/mg protein. Radioiodination of the purified enzyme followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed a single band at 69,000 Da. Kinetic analyses of the beta 6GlcNAc transferase-catalyzed reaction showed an ordered sequential mechanism in which UDP-GlcNAc binds to the enzyme first and UDP is released last. The Km values for UDP-GlcNAc and Gal beta 1-3GalNAc alpha Bzl were 0.36 and 0.14 mM, respectively. Acceptor competition studies showed that the purified beta 6GlcNAc transferase can use core 1 and core 3 mucin oligosaccharides as well as GlcNAc beta 1-3Gal beta R as acceptor substrates. Proton NMR analyses of the three products demonstrated that GlcNAc was added in a beta 1-6 linkage to the penultimate GalNAc or Gal, suggesting that this enzyme is capable of synthesizing all beta 6GlcNAc structures found in mucin-type oligosaccharides

Accurate estimates of absolute left ventricular volumes from equilibrium radionuclide angiographic count data using a simple geometric attenuation correction

To simplify and clarify the methods of obtaining attenuation-corrected equilibrium radionuclide angiographic estimates of absolute left ventricular volumes, 27 patients who also had biplane contrast cineangiography were evaluated. Background-corrected left ventricular end-diastolic and end-systolic counts were obtained by semiautomated variable and hand-drawn regions of interest and were normalized to cardiac cycles processed, frame rate and blood sample counts. Blood sample counts were acquired on (d°) and at a distance (d′) from the collimator. A simple geometric attenuation correction was performed to obtain absolute left ventricular volume estimates.Using blood sample counts obtained at d° or d′, the attentuation.corrected radionuclide left ventricular end-diastolic volume estimates using both region of interest selection methods correlated with the cineangiographic end-diastolic volumes (r = 0.95 to 0.96). However, both mean radionuclide semiautomated variable left ventricular end-diastolic volumes (179 ± 100 [± 1 standard deviation] and 185 ± 102 ml, p < 0.001) were smaller than the average cineangiographic end-diastolic volume (217 ± 102 ml), and both mean hand-drawn left ventricular end-diastolic volumes (212 ± 104 and 220 ± 106 ml) did not differ from the average cineangiographic end-diastolic volume. Using the blood sample counts obtained at d° or d′, the attenuation-corrected radionuclide left ventricular end-systolic volume estimates using both region of interest selection methods correlated with the cineangiographic end-systolic volumes (r = 0.96 to 0.98). Also, using blood sample counts at d°, the mean radionuclide semiautomated variable left ventricular end-systolic volume (116 ± 98 ml, p < 0.05) was less than the average cineangiographic end-systolic volume (128 ± 98 ml), and the other radionuclide end-systolic volumes did not differ from the average cineangiographic end-systolic volume.Therefore, it is concluded that: 1) a simple geometric attenuation-correction of radionuclide left ventricular end-diastolic and end-systolic count data provides accurate estimates of biplane cineangiographic end-diastolic and end-systolic volumes; and 2) the hand-drawn region of interest selection method, unlike the semiautomated variable method that underestimates end-diastolic and end-systolic volumes, provides more accurate estimates of biplane cineangiographic left ventricular volumes irrespective of the distance blood sample counts are acquired from the collimator