The chondroitin sulfate proteoglycan versican is important for embryonic development and several human disorders. The versican V1 splice isoform is widely expressed and cleaved by ADAMTS proteases at ...a well-characterized site, Glu441-Ala442. Since ADAMTS proteases cleave the homologous proteoglycan aggrecan at multiple sites, we hypothesized that additional cleavage sites existed within versican. We report a quantitative label-free approach that ranks abundance of liquid chromatography-tandem mass spectrometry (LC-MS/MS)-identified semi-tryptic peptides after versican digestion by ADAMTS1, ADAMTS4 and ADAMTS5 to identify site-specific cleavages. Recombinant purified versican V1 constructs were digested with the recombinant full-length proteases, using catalytically inactive mutant proteases in control digests. Semi-tryptic peptide abundance ratios determined by LC-MS/MS in ADAMTS:control digests were compared to the mean of all identified peptides to obtain a z-score by which outlier peptides were ranked, using semi-tryptic peptides identifying Glu441 -Ala442 cleavage as the benchmark. Tryptic peptides with higher abundance in control digests supported cleavage site identification. We identified several novel cleavage sites supporting the ADAMTS1/4/5 cleavage site preference for a P1-Glu residue in proteoglycan substrates. Digestion of proteins in vitro and application of this z-score approach is potentially widely applicable for mapping protease cleavage sites using label-free proteomics.
Versican abundance and turnover are relevant to the pathogenesis of several human disorders. Versican is cleaved by A Disintegrin-like And Metalloprotease with Thrombospondin type 1 motifs (ADAMTS) family members at Glu441-Ala442, generating a bioactive proteoform called versikine, but additional cleavage sites and the site-specificity of individual ADAMTS proteases is unexplored. Here, we used a label-free proteomics strategy to identify versican cleavage sites for 3 ADAMTS proteases, applying a novel z-score-based statistical approach to compare the protease digests of versican to controls (digests with inactive protease) using the known protease cleavage site as a benchmark. We identified 21 novel cleavage sites that had a comparable z-score to the benchmark. Given the functional significance of versikine, they represent potentially significant cleavages and helped to refine a substrate site preference for each protease.The z-score approach is potentially widely applicable for discovery of site-specific cleavages within an purified protein or small ensemble of proteins using any protease.
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•Versican proteolysis is biomedically significant, but few cleavage sites are known.•A novel z-score-based label-free proteomics method was used to define new cleavages.•Cleavage of two versican constructs by three ADAMTS proteases was tested.•A well-characterized versican cleavage site at Glu441-Ala442 was the benchmark.•Novel cleavage sites for ADAMTS1, ADAMTS4 and ADAMTS5 were identified.
The field of medical and population genetics in stroke is moving at a rapid pace and has led to unanticipated opportunities for discovery and clinical applications. Genome-wide association studies ...have highlighted the role of specific pathways relevant to etiologically defined subtypes of stroke and to stroke as a whole. They have further offered starting points for the exploration of novel pathways and pharmacological strategies in experimental systems. Mendelian randomization studies continue to provide insights in the causal relationships between exposures and outcomes and have become a useful tool for predicting the efficacy and side effects of drugs. Additional applications that have emerged from recent discoveries include risk prediction based on polygenic risk scores and pharmacogenomics. Among the topics currently moving into focus is the genetics of stroke outcome. While still at its infancy, this field is expected to boost the development of neuroprotective agents. We provide a brief overview on recent progress in these areas.
Response inhibition is essential for navigating everyday life. Its derailment is considered integral to numerous neurological and psychiatric disorders, and more generally, to a wide range of ...behavioral and health problems. Response-inhibition efficiency furthermore correlates with treatment outcome in some of these conditions. The stop-signal task is an essential tool to determine how quickly response inhibition is implemented. Despite its apparent simplicity, there are many features (ranging from task design to data analysis) that vary across studies in ways that can easily compromise the validity of the obtained results. Our goal is to facilitate a more accurate use of the stop-signal task. To this end, we provide 12 easy-to-implement consensus recommendations and point out the problems that can arise when they are not followed. Furthermore, we provide user-friendly open-source resources intended to inform statistical-power considerations, facilitate the correct implementation of the task, and assist in proper data analysis.
A comprehensive three‐dimensional fully coupled thermo‐electro‐mechanical finite element framework is developed for modeling spark plasma sintering (SPS). The finite element model is applied to the ...simulation of spark plasma processing with four different tooling sizes and various temperature regimes. The comparison of modeling and experimental results shows that the model is reliable for qualitative predictions of the densification behavior and of the grain growth in powder specimens subjected to SPS with a given temperature regime. The conducted modeling indicates the possibility of changing the heating pattern of the specimen (warmer central areas of the specimen's volume and cooler outside areas or vice versa) depending on the size of the tooling. High heating rates and large specimen sizes elevate the temperature and, in turn, material structure gradients during SPS processing. The obtained results suggest that the industrial implementation of SPS techniques should be based on the predictive capability of reliable modeling approaches.
The international development community is off-track from meeting targets for alleviating global malnutrition. Meanwhile, there is growing consensus across scientific disciplines that fish plays a ...crucial role in food and nutrition security. However, this ‘fish as food’ perspective has yet to translate into policy and development funding priorities. We argue that the traditional framing of fish as a natural resource emphasizes economic development and biodiversity conservation objectives, whereas situating fish within a food systems perspective can lead to innovative policies and investments that promote nutrition-sensitive and socially equitable capture fisheries and aquaculture. This paper highlights four pillars of research needs and policy directions toward this end. Ultimately, recognizing and working to enhance the role of fish in alleviating hunger and malnutrition can provide an additional long-term development incentive, beyond revenue generation and biodiversity conservation, for governments, international development organizations, and society more broadly to invest in the sustainability of capture fisheries and aquaculture.
The genetic basis of lacunar stroke is poorly understood, with a single locus on 16q24 identified to date. We sought to identify novel associations and provide mechanistic insights into the disease.
...We did a pooled analysis of data from newly recruited patients with an MRI-confirmed diagnosis of lacunar stroke and existing genome-wide association studies (GWAS). Patients were recruited from hospitals in the UK as part of the UK DNA Lacunar Stroke studies 1 and 2 and from collaborators within the International Stroke Genetics Consortium. Cases and controls were stratified by ancestry and two meta-analyses were done: a European ancestry analysis, and a transethnic analysis that included all ancestry groups. We also did a multi-trait analysis of GWAS, in a joint analysis with a study of cerebral white matter hyperintensities (an aetiologically related radiological trait), to find additional genetic associations. We did a transcriptome-wide association study (TWAS) to detect genes for which expression is associated with lacunar stroke; identified significantly enriched pathways using multi-marker analysis of genomic annotation; and evaluated cardiovascular risk factors causally associated with the disease using mendelian randomisation.
Our meta-analysis comprised studies from Europe, the USA, and Australia, including 7338 cases and 254 798 controls, of which 2987 cases (matched with 29 540 controls) were confirmed using MRI. Five loci (ICA1L-WDR12-CARF-NBEAL1, ULK4, SPI1-SLC39A13-PSMC3-RAPSN, ZCCHC14, ZBTB14-EPB41L3) were found to be associated with lacunar stroke in the European or transethnic meta-analyses. A further seven loci (SLC25A44-PMF1-BGLAP, LOX-ZNF474-LOC100505841, FOXF2-FOXQ1, VTA1-GPR126, SH3PXD2A, HTRA1-ARMS2, COL4A2) were found to be associated in the multi-trait analysis with cerebral white matter hyperintensities (n=42 310). Two of the identified loci contain genes (COL4A2 and HTRA1) that are involved in monogenic lacunar stroke. The TWAS identified associations between the expression of six genes (SCL25A44, ULK4, CARF, FAM117B, ICA1L, NBEAL1) and lacunar stroke. Pathway analyses implicated disruption of the extracellular matrix, phosphatidylinositol 5 phosphate binding, and roundabout binding (false discovery rate <0·05). Mendelian randomisation analyses identified positive associations of elevated blood pressure, history of smoking, and type 2 diabetes with lacunar stroke.
Lacunar stroke has a substantial heritable component, with 12 loci now identified that could represent future treatment targets. These loci provide insights into lacunar stroke pathogenesis, highlighting disruption of the vascular extracellular matrix (COL4A2, LOX, SH3PXD2A, GPR126, HTRA1), pericyte differentiation (FOXF2, GPR126), TGF-β signalling (HTRA1), and myelination (ULK4, GPR126) in disease risk.
British Heart Foundation.
Gilteritinib is a potent and selective
kinase inhibitor with single-agent clinical efficacy in relapsed/refractory
-mutated acute myeloid leukemia (AML). In this context, however, gilteritinib is not ...curative, and response duration is limited by the development of secondary resistance. To evaluate resistance mechanisms, we analyzed baseline and progression samples from patients treated on clinical trials of gilteritinib. Targeted next-generation sequencing at the time of AML progression on gilteritinib identified treatment-emergent mutations that activate RAS/MAPK pathway signaling, most commonly in
or
Less frequently, secondary
-F691L gatekeeper mutations or
fusions were identified at progression. Single-cell targeted DNA sequencing revealed diverse patterns of clonal selection and evolution in response to FLT3 inhibition, including the emergence of
mutations in
-mutated subclones, the expansion of alternative wild-type
subclones, or both patterns simultaneously. These data illustrate dynamic and complex changes in clonal architecture underlying response and resistance to mutation-selective tyrosine kinase inhibitor therapy in AML. SIGNIFICANCE: Comprehensive serial genotyping of AML specimens from patients treated with the selective FLT3 inhibitor gilteritinib demonstrates that complex, heterogeneous patterns of clonal selection and evolution mediate clinical resistance to tyrosine kinase inhibition in
-mutated AML. Our data support the development of combinatorial targeted therapeutic approaches for advanced AML.
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BACKGROUND:Cytokines and growth factors have been implicated in the initiation and propagation of vascular disease. Observational studies have shown associations of their circulating levels with ...stroke. Our objective was to explore whether genetically determined circulating levels of cytokines and growth factors are associated with stroke and its etiologic subtypes by conducting a 2-sample Mendelian randomization (MR) study.
METHODS:Genetic instruments for 41 cytokines and growth factors were obtained from a genome-wide association study of 8293 healthy adults. Their associations with stroke and stroke subtypes were evaluated in the MEGASTROKE genome-wide association study data set (67 162 cases; 454 450 controls) applying inverse variance–weighted meta-analysis, weighted-median analysis, Mendelian randomization–Egger regression, and multivariable Mendelian randomization. The UK Biobank cohort was used as an independent validation sample (4985 cases; 364 434 controls). Genetic instruments for monocyte chemoattractant protein-1 (MCP-1/CCL2) were further tested for association with etiologically related vascular traits by using publicly available genome-wide association study data.
RESULTS:Genetic predisposition to higher MCP-1 levels was associated with higher risk of any stroke (odds ratio OR per 1 SD increase, 1.06; 95% CI, 1.02–1.09; P=0.0009), any ischemic stroke (OR, 1.06; 95% CI, 1.02–1.10; P=0.002), large-artery stroke (OR, 1.19; 95% CI, 1.09–1.30; P=0.0002), and cardioembolic stroke (OR, 1.14; 95% CI, 1.06–1.23; P=0.0004), but not with small-vessel stroke or intracerebral hemorrhage. The results were stable in sensitivity analyses and remained significant after adjustment for cardiovascular risk factors. Analyses in the UK Biobank showed similar associations for available phenotypes (any strokeOR, 1.08; 95% CI, 0.99–1.17; P=0.09; any ischemic strokeOR, 1.07; 95% CI, 0.97–1.18; P=0.17). Genetically determined higher MCP-1 levels were further associated with coronary artery disease (OR, 1.04; 95% CI, 1.00–1.08; P=0.04) and myocardial infarction (OR, 1.05; 95% CI, 1.01–1.09; P=0.02), but not with atrial fibrillation. A meta-analysis of observational studies showed higher circulating MCP-1 levels in patients with stroke in comparison with controls.
CONCLUSIONS:Genetic predisposition to elevated circulating levels of MCP-1 is associated with higher risk of stroke, in particular with large-artery stroke and cardioembolic stroke. Whether targeting MCP-1 or its receptors can lower stroke incidence requires further study.
ABSTRACT
We present the measurement of the Hubble constant, H0, with three strong gravitational lens systems. We describe a blind analysis of both PG 1115+080 and HE 0435−1223 as well as an extension ...of our previous analysis of RXJ 1131−1231. For each lens, we combine new adaptive optics (AO) imaging from the Keck Telescope, obtained as part of the SHARP (Strong-lensing High Angular Resolution Programme) AO effort, with Hubble Space Telescope (HST) imaging, velocity dispersion measurements, and a description of the line-of-sight mass distribution to build an accurate and precise lens mass model. This mass model is then combined with the COSMOGRAIL-measured time delays in these systems to determine H0. We do both an AO-only and an AO + HST analysis of the systems and find that AO and HST results are consistent. After unblinding, the AO-only analysis gives $H_{0}=82.8^{+9.4}_{-8.3}~\rm km\, s^{-1}\, Mpc^{-1}$ for PG 1115+080, $H_{0}=70.1^{+5.3}_{-4.5}~\rm km\, s^{-1}\, Mpc^{-1}$ for HE 0435−1223, and $H_{0}=77.0^{+4.0}_{-4.6}~\rm km\, s^{-1}\, Mpc^{-1}$ for RXJ 1131−1231. The joint AO-only result for the three lenses is $H_{0}=75.6^{+3.2}_{-3.3}~\rm km\, s^{-1}\, Mpc^{-1}$. The joint result of the AO + HST analysis for the three lenses is $H_{0}=76.8^{+2.6}_{-2.6}~\rm km\, s^{-1}\, Mpc^{-1}$. All of these results assume a flat Λ cold dark matter cosmology with a uniform prior on Ωm in 0.05, 0.5 and H0 in 0, 150 $\rm km\, s^{-1}\, Mpc^{-1}$. This work is a collaboration of the SHARP and H0LiCOW teams, and shows that AO data can be used as the high-resolution imaging component in lens-based measurements of H0. The full time-delay cosmography results from a total of six strongly lensed systems are presented in a companion paper.
Wearable Spectroradiometer for Dosimetry Chmielinski, Maximilian J; Cohen, Martin A; Yost, Michael G ...
Sensors (Basel, Switzerland),
11/2022, Letnik:
22, Številka:
22
Journal Article
Recenzirano
Odprti dostop
Available wearable dosimeters suffer from spectral mismatch during the measurement of broadband UV and visible radiation in environments that receive radiation from multiple sources emitting ...differing spectra. We observed this type of multi-spectra environment in all five Washington State cannabis farms visited during a field study investigating worker exposure to ultraviolet radiation in 2018. Spectroradiometers do not suffer from spectral mismatch in these environments, however, an extensive literature review conducted at the time of writing did not identify any spectroradiometers that were directly deployable as wearable dosimetry devices. To close this research gap, we developed a microcontroller system and platform that allows for researchers to mount and deploy the Ocean Insight Flame-S Spectroradiometer as a wearable device for measurement of UV and visible wavelengths (300 to 700 nm). The platform validation consisted of comparing measurements taken under platform control with measurements taken with the spectrometer controlled by a personal computer running the software provided by the spectroradiometer manufacturer. Three Mann-Whitney U-Tests (two-tailed, 95% CI), one for each intensity condition, compared the central tendency between the total spectral power (TSP), the integral of a spectrum measurement, measured under both control schemas. An additional analysis of per pixel agreement and overall platform stability was performed. The three Mann-Whitney tests returned no significant difference between the set of TSPs for each filter condition. These results suggest that the spectroradiometer takes measurements of equivalent accuracy under both control schemas, and can be deployed as a wearable device for the measurement of wavelength resolved UV and visible radiation.
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