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An improved method for isolation of RNA from bone

<p>Abstract</p> <p>Background</p> <p>Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis using the currently available approaches.</p> <p>Results</p> <p>We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the phenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This single step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from 6.7 to 9.2.</p> <p>Conclusions</p> <p>Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue while facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA obtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray, and next generation sequencing applications.</p

Glucocorticoid therapy in ANCA Vasculitis - using the Glucocorticoid Toxicity Index as an outcome measure

Background Anti-neutrophil cytoplasm antibodies (ANCA) associated vasculitis (AAV) is an autoimmune disease. Induction remission and maintenance treatment typically includes high dose, tapering glucocorticoids (GC) in addition to other immunosuppressive medication. The use of Glucocorticoid Toxicity Index (GTI), provides a global, quantifiable assessment tool in which clinicians can assess GC associated morbidity. Recent trials in AAV have exposed the need for systemic assessment of GC burden. In this small cohort study, we look to address these issues and the justification of newer GC sparing agents such as C5a inhibitors. Methods A retrospective cohort study of 43 patients with biopsy AAV was constructed from a single centre between 2012 to 2016 and followed up for 48 months. The GTI table made up of adverse features used to quantify patients GC toxicity. Electronic patient records were reviewed and scores calculated according to published methods. GTI scores were compared with cumulative steroid doses at separate intervals as well as incidences of adverse features in relation to the treatment timeline. Results The mean age was 65.9 (± 11.06) years and treatment regimens consisted of glucocorticoids alongside cyclophosphamide or rituximab. Our results showed statistical significance in the association of cumulative GC doses and GTI scores (p=0.008, 95% CI, 1.31 to 8.05). Adverse features relating to mood disturbance and GC induced psychosis occurred early, in contrast to adrenal insufficiency which typically presented later in the follow up. Infection related adverse events were consistent throughout. Conclusions We demonstrated that higher, cumulative doses of steroids in AAV lead to worse glucocorticoid related toxicity. Using the GTI creates potential to individualise and quantify the adverse effects patients experience as a result of GC treatment and permits more patient centred management. Whilst glucocorticoids remain the main adjunctive immunosuppression of AAV treatment, the narrow therapeutic window supports the need for GC-sparing treatments

A Pilot Study of Cultural-Based Service-Learning: What Did Undergraduate Students Learn from Elementary School Teachers in an Urban Public School District?

This article describes what undergraduate students learned from school teachers involved in a cultural based service-learning program. Forty-four undergraduate students enrolled in an educational psychology course at a private, metropolitan University were paired with 30 teachers and provided tutoring and mentoring activities to the children in their classrooms over the course of the semester at the West or East School located in Chester, Pennsylvania. The results from a preliminary survey indicate that undergraduate students increase their civic action attitudes, problem-solving skills, multigroup and ethnic identity, community self-efficacy, and academic and community engagement from the beginning to the end of the term. Undergraduate students acquired confidence in their communication, analytical, and problem-solving skills. Undergraduate students also developed their ethnic identity and made connections to the academic community and beyond to the broader University community. The results further indicate that the service context influences students’ critical thinking and social responsibility. Undergraduate students assigned to the West School had higher post service ratings of their application skills and community responsibility compared to those assigned to the East School. Teachers’ reports confirm that undergraduate students demonstrate cultural competence and community engagement in their service activities at both the West and East School

Differential Regulation of Neuropeptide Y in the Amygdala and Prefrontal Cortex during Recovery from Chronic Variable Stress

Accumulating evidence from clinical studies and pre-clinical animal models supports a role for neuropeptide Y (NPY) in adaptive emotional response following stress. The long-term impact of stress, particularly chronic stress, on availability, and function of resilience factors such as NPY may be critical to understanding the etiology of stress-related psychopathology. In these studies, we examined expression of NPY during recovery from a chronic variable stress (CVS) model of repetitive trauma in rats. Due to the importance of amygdala and prefrontal cortex in regulating emotional responses, we predicted chronic changes in NPY expression could contribute to persistent behavioral deficits seen in this model. Consistent with the hypothesis, ELISA for NPY peptide identified a significant reduction in NPY at the delayed (7 days) recovery time-point. Interestingly, a significant increase in prefrontal NPY was observed at the same recovery time-point. The mRNA expression for NPY was not changed in the amygdala or PFC, although there was a modest but not statistically significant increase in NPY mRNA at the delayed recovery time-point in the prefrontal cortex. The observed changes in NPY expression are consistent with maladaptive coping and enhanced emotionality, due to the nature of NPY signaling within these respective regions, and the nature of reciprocal connections between amygdala and prefrontal cortex

Portable, Low-Cost, Column Carbon Dioxide and Methane Measurements for Validating Satellite Observations in Remote Locations

We present a low-cost (~$10K per instrument), portable solution to ground-based validation of satellite observations for difficult to reach locations with precisions of 1 ppm XCO2 and 10 ppb XCH4 for hourly data products. While Total Carbon Column Observing Network (TCCON) is the gold-standard for ground validation, there are locations where a ground column validation data point would be useful but conditions are not conducive to a permanent station. Examples include wetlands, thawing permafrost, the tropics, the Amazon, sub-Saharan Africa, as well as locations without a power grid or with geopolitical conflict. In addition, the low-cost and portability mean a geographical region can be studied in depth with multiple instruments. This passive, sun-pointing instrument is a miniaturized, laser heterodyne radiometer (mini-LHR) that has been under development by our team since 2009. It can be operated either in tandem with AERONET (a global network of 500 instruments that measure aerosol optical depth), or as a stand-alone instrument with a low-cost (~$3K), light-weight sun tracker. One of the main benefits of the mini-LHR is that it can quickly reach remote locations and provide a validation measurement even if there is limited or no infrastructure at the site. The instrument weighs ~10 lbs, fits into a backpack, and is powered by two folding solar panels and a battery pack. In clear conditions, the instrument can be set-up in under an hour. Portability means that mini-LHRs can be easily moved for side-by-side comparisons with other mini-LHRs and with TCCON which simplifies assessing instrument bias as well as accuracy. Like TCCON, the mini-LHR points directly at the sun with a narrow field-of-view and is its insensitivity to cloud and aerosol scattering that is common to nadir-pointing passive satellite approaches. Here we present a collection of sample data sets to demonstrate performance from locations that vary in climate, altitude, solar zenith angle, hours of sun per day, etc., as well as data from side-by-side TCCON comparisons. Retrievals of CO2 and CH4 were completed using the NASA/Goddard's Planetary Spectrum Generator (PSG) that incorporates meteorological inputs from Modern-Era Retrospective analysis for Research and Applications, Version 2 (MERRA-2) data set

The ubiquitin ligase Siah2 regulates obesity-induced adipose tissue inflammation

© 2015 The Obesity Society. Objective Chronic, low-grade adipose tissue inflammation associated with adipocyte hypertrophy is an important link in the relationship between obesity and insulin resistance. Although ubiquitin ligases regulate inflammatory processes, the role of these enzymes in metabolically driven adipose tissue inflammation is relatively unexplored. Herein, the effect of the ubiquitin ligase Siah2 on obesity-related adipose tissue inflammation was examined. Methods Wild-type and Siah2KO mice were fed a low- or high-fat diet for 16 weeks. Indirect calorimetry, body composition, and glucose and insulin tolerance were assayed along with glucose and insulin levels. Gene and protein expression, immunohistochemistry, adipocyte size distribution, and lipolysis were also analyzed. Results Enlarged adipocytes in obese Siah2KO mice were not associated with obesity-induced insulin resistance. Proinflammatory gene expression, stress kinase signaling, fibrosis, and crown-like structures were reduced in the Siah2KO adipose tissue, and Siah2KO adipocytes were more responsive to insulin-dependent inhibition of lipolysis. Loss of Siah2 increased expression of PPARγ target genes involved in lipid metabolism and decreased expression of proinflammatory adipokines regulated by PPARγ. Conclusions Siah2 links adipocyte hypertrophy with adipocyte dysfunction and recruitment of proinflammatory immune cells to adipose tissue. Selective regulation of PPARγ activity is a Siah2-mediated mechanism contributing to obesity-induced adipose tissue inflammation

Diels-Alder-based thermo-reversibly crosslinked polymers:Interplay of crosslinking density, network mobility, kinetics and stereoisomerism

Polymers crosslinked through thermo-reversible furan/maleimide Diels-Alder chemistry have been widely explored, since they stand as an ingenious design for reprocessable and self-healing thermosets and elastomers. For these polymeric products, crosslinking density plays a key role on the polymer thermo-reversibility. However, how this degree of network interconnectivity influences the kinetics of thermal reversibility has not yet been addressed. In order to tackle this problem, furan-grafted polyketones crosslinked by a bi-functional maleimide were prepared with different ratios between maleimide and furan groups. The thermo-reversible dynamics of the prepared polymers were then studied by rheology and differential scanning calorimetry. Here we show that, the thermo-reversible process occurs faster and at lower temperatures in polymers with lower crosslinking densities. Network mobility is responsible for this effect. It allows the formulations to rearrange their polymer network differently through the heating-cooling cycles. The results also indicate that the crosslinking density rather than the stereoisomerism of the Diels-Alder adducts plays a larger role in the reversible behavior of the system. Additionally, the thermo-reversible features of the polymer were shown to be dependent on its thermal history. This work impacts the development of reprocessable and self-healing crosslinked polymers, and the design of the corresponding reprocessing and healing procedures