Rapid pre-gel visualization of proteins with mass spectrometry compatibility

Despite all of the prophecies of doom, gel electrophoresis is still prevalent in modern proteomic workflows. However, the currently used protein staining methods represent a serious bottleneck for a quick subsequent protein analysis using mass spectrometry. Substituting traditional protein stains by pre-gel derivatization with visible and mass spectrometry compatible reagents eliminates several processing steps and drastically reduces the sample preparation time. A defined chemistry permits seamless integration of such covalent protein staining methods into standardized bioinformatic pipelines. Using Uniblue A we could covalently stain simple to complex protein samples within 1 minute. Protein profiles on the gels were not compromised and MS/MS based sequence coverages higher than 80% could be obtained. In addition, the visual tracking of covalently stained proteins and peptides facilitates method development and validation. Altogether, this new chemo-proteomic approach enables true "at-line" analysis of proteins

Latent Growth Modeling of the Attitudes of Non-Muslims Towards Muslims in College: A Longitudinal Analysis

Attitudes toward Muslims are at the core of addressing the anti-Muslim environments Muslims encounter in college and society writ large. Using longitudinal data, this study shows that colleges can spur the development of appreciative attitudes toward Muslims by non-Muslim students when they engage in two or more informal or formal social engagements across religious and spiritual identity differences. Institutional campus culture is also meaningfully connected to Muslim appreciation. Implications for research, policy, and practice are discussed

Application of Differential Pulse Voltammetry to Determine the Efficiency of Stripping Tocopherols from Commercial Fish Oil

There has been an increase in the use of electrochemical methods for monitoring antioxidant levels in a variety of disciplines due to the sensitivity, low detection limits, ease of use, low cost and rapid analysis time offered by these techniques. One technique that has received specific attention is differential pulse voltammetry. We describe a novel application of differential pulse voltammetry to quantitatively and qualitatively determine the efficiency of removing tocopherols from commercial fish oil via column chromatographic separation. The relative limits of detection and quantitation of differential pulse voltammetry are compared to HPLC for determining the removal of tocopherols from commercial fish oil. It was determined that differential pulse voltammetry can monitor the separation of commercially added antioxidants from the bulk sample via a decrease in antioxidant oxidation currents. Furthermore, the limits of detection and quantitation were found to be comparable with values obtained using HPLC for tocopherol identification and quantitation.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141339/1/aocs0527.pd

Electron tunneling through sensitizer wires bound to proteins

We report a quantitative theoretical analysis of long-range electron transfer through sensitizer wires bound in the active-site channel of cytochrome P450cam. Each sensitizer wire consists of a substrate group with high binding affinity for the enzyme active site connected to a ruthenium-diimine through a bridging aliphatic or aromatic chain. Experiments have revealed a dramatic dependence of electron transfer rates on the chemical composition of both the bridging group and the substrate. Using combined molecular dynamics simulations and electronic coupling calculations, we show that electron tunneling through perfluorinated aromatic bridges is promoted by enhanced superexchange coupling through virtual reduced states. In contrast, electron flow through aliphatic bridges occurs by hole-mediated superexchange. We have found that a small number of wire conformations with strong donor–acceptor couplings can account for the observed electron tunneling rates for sensitizer wires terminated with either ethylbenzene or adamantane. In these instances, the rate is dependent not only on electronic coupling of the donor and acceptor but also on the nuclear motion of the sensitizer wire, necessitating the calculation of average rates over the course of a molecular dynamics simulation. These calculations along with related recent findings have made it possible to analyze the results of many other sensitizer-wire experiments that in turn point to new directions in our attempts to observe reactive intermediates in the catalytic cycles of P450 and other heme enzymes

HIPGEN: a randomized, multicentre phase III study using intramuscular PLacenta-eXpanded stromal cells therapy for recovery following hip fracture arthroplasty : a study design

Aims The aim of the HIPGEN consortium is to develop the first cell therapy product for hip fracture patients using PLacental-eXpanded (PLX-PAD) stromal cells. Methods HIPGEN is a multicentre, multinational, randomized, double-blind, placebo-controlled trial. A total of 240 patients aged 60 to 90 years with low-energy femoral neck fractures (FNF) will be allocated to two arms and receive an intramuscular injection of either 150 × 106 PLX-PAD cells or placebo into the medial gluteal muscle after direct lateral implantation of total or hemi hip arthroplasty. Patients will be followed for two years. The primary endpoint is the Short Physical Performance Battery (SPPB) at week 26. Secondary and exploratory endpoints include morphological parameters (lean body mass), functional parameters (abduction and handgrip strength, symmetry in gait, weightbearing), all-cause mortality rate and patient-reported outcome measures (Lower Limb Measure, EuroQol five-dimension questionnaire). Immunological biomarker and in vitro studies will be performed to analyze the PLX-PAD mechanism of action. A sample size of 240 subjects was calculated providing 88% power for the detection of a 1 SPPB point treatment effect for a two-sided test with an α level of 5%. Conclusion The HIPGEN study assesses the efficacy, safety, and tolerability of intramuscular PLX-PAD administration for the treatment of muscle injury following arthroplasty for hip fracture. It is the first phase III study to investigate the effect of an allogeneic cell therapy on improved mobilization after hip fracture, an aspect which is in sore need of addressing for the improvement in standard of care treatment for patients with FNF

An In-vivo 1H-MRS short-echo time technique at 7T: Quantification of metabolites in chronic multiple sclerosis and neuromyelitis optica brain lesions and normal appearing brain tissue

Highlights NAAG likely contributes to the total NAA differences between multiple sclerosis lesion and normal appearing brain tissue. myo-Inositol was not shown to be different between chronic AQP4Ab-NMOSD brain lesions and normal appearing brain tissue. An optimised MRS methodology is described, using 7T field strength and correcting for tissue T2 water relaxion differences. 7-tesla MRS profiles of chronic brain lesions and normal appearing white matter are presented for MS and AQP4Ab-NMOSD. Abstract Magnetic Resonance Spectroscopy (MRS) allows for the non-invasive quantification of neurochemicals and has the potential to differentiate between the pathologically distinct diseases, multiple sclerosis (MS) and AQP4Ab-positive neuromyelitis optica spectrum disorder (AQP4Ab-NMOSD). In this study we characterised the metabolite profiles of brain lesions in 11 MS and 4 AQP4Ab-NMOSD patients using an optimised MRS methodology at ultra-high field strength (7T) incorporating correction for T2 water relaxation differences between lesioned and normal tissue. MS metabolite results were in keeping with the existing literature: total N-acetylaspartate (NAA) was lower in lesions compared to normal appearing brain white matter (NAWM) with reciprocal findings for myo-Inositol. An unexpected subtlety revealed by our technique was that total NAA differences were likely driven by NAA-glutamate (NAAG), a ubiquitous CNS molecule with functions quite distinct from NAA though commonly quantified together with NAA in MRS studies as total NAA. Surprisingly, AQP4Ab-NMOSD showed no significant differences for total NAA, NAA, NAAG or myo-Inositol between lesion and NAWM sites, nor were there any differences between MS and AQP4Ab-NMOSD for a priori hypotheses. Post-hoc testing revealed a significant correlation between NAWM Ins:NAA and disability (as measured by EDSS) for disease groups combined, driven by the AP4Ab-NMOSD group. Utilising an optimised MRS methodology, our study highlights some under-explored subtleties in MRS profiles, such as the absence of myo-Inositol concentration differences in AQP4Ab-NMOSD brain lesions versus NAWM and the potential influence of NAAG differences between lesions and normal appearing white matter in MS

Assessing the response of plant flavonoids to UV radiation : an overview of appropriate techniques

Flavonoids are a large group of plant secondary metabolites that are present in most plants, and are vital for plant growth, development and protection. Among the many functions of these compounds is their contribution to stress amelioration. The accurate identification and quantification of total or individual flavonoids in plants exposed to stressful conditions (e.g. ultraviolet radiation) is challenging due to their structural diversity. The present review provides the up to date knowledge and highlights trends in plant flavonoid analysis. The review covers all steps from the field to the laboratory, focussing on UV-B effects on flavonoids, and identifying critical issues concerning sample collection, pre-treatment, extraction techniques and quantitative or qualitative analysis. A well-planned sampling and sample prehandling strategy is vital when capturing organ, tissue and developmental-stage dependent changes in flavonoids, as well as the dynamic changes due to time of UV-exposure and diurnal or seasonal parameters. A range of advanced extraction and purification techniques can facilitate the quantitative transfer of flavonoids to solvents. The advantages and disadvantages of analytical methods, including chromogenic assays, liquid and thin-layer chromatography, mass spectrometry, nuclear magnetic resonance detection, and non-destructive in situ fluorescent analysis need to be consciously evaluated in the context of the specific biological question posed. Thus, no one method can be applied to every single study of flavonoid. The message of this review is that researchers will need to carefully consider the biological process that they intend to study, and select an analytical method that optimally matches their specific objectives.Peer reviewe

Snowmelt Timing Regulates Community Composition, Phenology, and Physiological Performance of Alpine Plants

The spatial patterning of alpine plant communities is strongly influenced by the variation in physical factors such as temperature and moisture, which are strongly affected by snow depth and snowmelt patterns. Earlier snowmelt timing and greater soil-moisture limitations may favor wide-ranging species adapted to a broader set of ecohydrological conditions than alpine-restricted species. We asked how plant community composition, phenology, plant water relations, and photosynthetic gas exchange of alpine-restricted and wide-ranging species differ in their responses to a ca. 40-day snowmelt gradient in the Colorado Rocky Mountains (Lewisia pygmaea, Sibbaldia procumbens, and Hymenoxys grandiflora were alpine-restricted and Artemisia scopulorum, Carex rupestris, and Geum rossii were wide-ranging species). As hypothesized, species richness and foliar cover increased with earlier snowmelt, due to a greater abundance of wide-ranging species present in earlier melting plots. Flowering initiation occurred earlier with earlier snowmelt for 12 out of 19 species analyzed, while flowering duration was shortened with later snowmelt for six species (all but one were wide-ranging species). We observed >50% declines in net photosynthesis from July to September as soil moisture and plant water potentials declined. Early-season stomatal conductance was higher in wide-ranging species, indicating a more competitive strategy for water acquisition when soil moisture is high. Even so, there were no associated differences in photosynthesis or transpiration, suggesting no strong differences between these groups in physiology. Our findings reveal that plant species with different ranges (alpine-restricted vs. wide-ranging) could have differential phenological and physiological responses to snowmelt timing and associated soil moisture dry-down, and that alpine-restricted species’ performance is more sensitive to snowmelt. As a result, alpine-restricted species may serve as better indicator species than their wide-ranging heterospecifics. Overall, alpine community composition and peak % cover are strongly structured by spatio-temporal patterns in snowmelt timing. Thus, near-term, community-wide changes (or variation) in phenology and physiology in response to shifts in snowmelt timing or rates of soil dry down are likely to be contingent on the legacy of past climate on community structure

Oxidation of Fish Oil Oleogels Formed by Natural Waxes in Comparison With Bulk Oil

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143663/1/ejlt201700378.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/143663/2/ejlt201700378_am.pd

Sectioned or whole otoliths? A global review of hard structure preparation techniques used in ageing sparid fishes

While otoliths are considered the most reliable structure to accurately age fish, a variety of otolith preparation techniques are available, which have consequences on the otolith’s optical properties and therefore interpretation of growth bands. Recently, numerous studies from a variety of authors have criticised the use of whole otoliths in ageing sparids with sectioned otoliths subsequently acknowledged as the most reliable preparation technique. Despite this criticism; ageing data is still being generated from whole otoliths and other unreliable structures such as scales. In an attempt to understand the severity of this issue we conducted a global literature review of otolith preparation protocols used for sparids. We identified global spatial inconsistencies in otolith preparation techniques with some regions predominately using methods other than sectioned otoliths to age sparids. The review highlights the need for a standardisation of otolith preparation methods and a move towards the use of sectioned otoliths, or at least valid support where alternative structures or preparation techniques are used. Given that large numbers of studies have been conducted on whole otoliths in certain regions, it may be necessary to revaluate the existing growth parameters to ensure that accurate information is incorporated into management structures