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Abstract MicroRNAs (miRNAs) are short non-coding RNAs that are involved in post-transcriptional control of gene expression and might be used as biomarkers for diabetes-related complications. The aim of this case–control study was to explore potential differences in circulating miRNAs in young individuals with long-duration type 1 diabetes (T1D) compared to healthy controls, and how identified miRNAs are expressed across different tissues. Twelve adolescents, age 15.0–17.9 years, with T1D duration of more than 8 years (mean 11.1 years), were enrolled from the Swedish diabetes quality registry. An age-matched control group was recruited. Circulating miRNAs (n = 187) were analyzed by quantitative PCR. We observed that 27 miRNAs were upregulated and one was downregulated in T1D. Six of these miRNAs were tissue-enriched (blood cells, gastrointestinal, nerve, and thyroid tissues). Six miRNAs with the largest difference in plasma, five up-regulated (hsa-miR-101-3p, hsa-miR-135a-5p, hsa-miR-143-3p, hsa-miR-223-3p and hsa-miR-410-3p (novel for T1D)) and one down-regulated (hsa-miR-495-3p), with P-values below 0.01, were selected for further in-silico analyses. AKT1, VEGFA and IGF-1 were identified as common targets. In conclusion, 28 of the investigated miRNAs were differently regulated in long-duration T1D in comparison with controls. Several associations with cancer were found for the six miRNAs with the largest difference in plasma.

Efforts towards climate neutrality in Europe must prioritise manufacturing industries, particularly the energy-intensive industry (EII) subsectors. This work proposes a novel approach to assessing transformation options for EII subsectors. At the center of this approach we position a potential analysis of technologies’ impact on subsector decarbonisation—an approach only known so far from the investigation of renewable energy potentials. These so-called technical climate neutrality potentials, supplemented by a set of indicators taking into account energy consumption, capital and operational expenditures, and GHG taxation programs per technology and subsector, enable cross-sector comparisons. The indicators allow the reader to compare the impact on GHG emission mitigation, energy demand, and cost for every considered technology. At the same time, we keep an open mind regarding combinations of technological solutions in the overall energy system. This ensures that the technology pathways with the greatest climate neutrality potential are easily identified. These focal points can subsequently serve in, e.g., narrative-driven scenario analyses to define comprehensive guides for action for policymakers. A case study of Austria for the proposed potential analysis demonstrates that bio-CH4 and electrolysis-derived H2 are the most economical green gases, but GHG certificate costs will be necessary for cost-competitiveness in high-temperature applications. Electrification offers advantages over conventional technologies and CO2-neutral gas alternatives in low-to-mid temperature ranges. Under the given assumptions, including GHG emission certificate costs of 250 EUR/t CO2, alternative technologies in the identified climate neutrality pathways can operate at total annual costs comparable to conventional fossil-based equivalents.

Two long-lasting thunderstorm ground enhancement (TGE) events were registered at the Milešovka meteorological observatory in Czechia (50.55∘ N, 13.93∘ E; 837 m altitude) on 23 April 2018, during linearly organized thunderstorms. Two intervals of increased photon counts were detected by a plastic scintillator, respectively lasting 70 and 25 min and reaching 31 % and 48 % above the background radiation levels. Using numerical simulations, we verified that the observed increases in count rates are consistent with the energy spectrum of previously observed TGEs. We investigated the relevant data from a suite of meteorological instruments, a Ka-band cloud radar, an electric field mill, and a broadband electromagnetic receiver, all placed at the Milešovka observatory, in order to analyse the context in which these unique continental TGEs occurred at an exceptionally low altitude. The onset of the TGEs preceded the onset of precipitation by 10 and 3 min, respectively, for the two events. Both this delayed rain arrival and an energy threshold of 6.5 MeV for registered particles clearly exclude the detection the decay products of the radon progeny washout during the TGE intervals. At the same time, the European lightning detection network EUCLID detected numerous predominantly negative intracloud lightning discharges at distances closer than 5 km from the particle detector, while the occurrence of cloud-to-ground discharges was suppressed. The cloud radar recorded presence of graupel below the melting level, and the composition of hydrometeors suggested good conditions for cloud electrification. The observed variations in the near-surface electric field were unusual, with very brief negative-electric-field excursions reaching −20 kV in a quick succession. At the same time, sub-microsecond unipolar pulses emitted by close corona discharges saturated the broadband magnetic loop antenna. All these measurements indicate that a strong lower positive-charge region was present inside the thundercloud. The bottom thundercloud dipole was probably responsible for acceleration of the seed electrons in the air. These seed electrons might originate in the secondary cosmic ray particles but could also come from a high concentration of radon in the air collected during the propagation of the convective system above the uranium-rich soils before the thunderstorms overpassed the Milešovka observatory.

Hypophosphatasia (HPP) is characterized by low activity of tissue nonspecific alkaline phosphatase (TNSALP). The enzyme replacement therapy asfotase alfa has been approved for childhood-onset forms of HPP. MicroRNAs (miRNAs) have emerged as a novel disease biomarker, with potential application in therapy monitoring. Circulating miRNAs were analyzed at baseline, months 1, 2, 4, and 16 in a 49-yr-old woman with childhood-onset HPP, chronic musculoskeletal pain, and non-traumatic fractures prior to enzyme replacement therapy. Serum RNA was extracted and sequenced using miRNeasy Mini Kit (Qiagen, Germany), RealSeq Biosciences Kit (Santa Cruz, US) together with miND spike-in control kit (TAmiRNA, Austria) and Illumina NovaSeq 6000 SP1 flow cell (San Diego, US). Brief Pain Inventory Severity and Interference scores (BPI-S/BPI-I), fatigue severity scale (FSS), Patient Global Impression of Improvement (PGI-I), Western Ontario and McMaster university hip disability and osteoarthritis outcome score (WOMAC), fibromyalgia impact questionnaire (FIQ), 6-Minute Walking Test (6-MWT), chair-rise-test (CRT), and handgrip dynamometry (HD) were performed at baseline and different timepoints during the therapy. Out of >800 screened, 84 miRNAs were selected based on differences in expression profiles between 24 HPP patients and 24 healthy controls. Six miRNAs showed a clear graphic trend and were up- or downregulated by ≥50% reads per million (rpm). These included hsa-let-7i-5p (+50%), hsa-miR-1-3p (−66.66%), hsa-miR-1294 (+63.63%), hsa-miR-206 (−85.57%), hsa-miR-375-3p (−71.43%), and hsa-miR-624-5p (+69.44%). hsa-miR-1-3p and hsa-miR-206 were identified as muscle-specific miRNAs. hsa-mir-375-3p, which negatively regulates osteogenesis, was significantly downregulated. In terms of patient-reported outcomes, BPI-S, BPI-I, FSS, PGI-I, WOMAC, and FIQ showed a reduction by −58.62%, −68.29%, −33.33%, −75.00%, −63.29%, and −43.02%, respectively. 6-MWT improved by +33.89% and CRT by −44.46%. Mean hand grip strength of the right/left hand measured by HD improved by +12.50% and + 23.53%, respectively. miRNA profile changes during the therapy with asfotase alfa, accompanying improvements in functionality tests and quality of life scores. Lay Summary: Hypophosphatasia (HPP) is a rare genetic bone disease characterized by low activity of an enzyme involved in mineralization of bony tissues, the nonspecific alkaline phosphatase (TNSALP). The enzyme replacement therapy with recombinant asfotase alfa has been approved recently. MicroRNAs (miRNAs) are small signaling molecules, which are involved in the regulation of gene expression and signaling pathways. They emerged as a novel disease biomarker, with potential application in therapy monitoring. We analyzed the levels of miRNAs of a 49-yr-old woman with childhood-onset HPP during the therapy. The patient suffered from musculoskeletal pain and non-traumatic fractures before the begin of enzyme replacement therapy. miRNAs in blood and clinical tests were performed at baseline and different timepoints. Blood levels of 6 miRNAs showed a clear graphic trend and were elevated or lowered by ≥50% units (reads per million) during the therapy. Patient-reported questionnaires showed a reduction of at least a third, reflecting clinical improvement of symptoms during the therapy. The scores of 6-MWT and CRT and hand grip also improved, signaling improvements in muscle functionality under asfotase alfa. miRNA profile changes during the therapy with asfotase alfa, accompanying improvements in functionality tests and quality of life scores. [ABSTRACT FROM AUTHOR]

Extracellular vesicles comprise an as yet inadequately investigated intercellular communication pathway in the field of early osteoarthritis. We hypothesised that the small non-coding RNA expression pattern in synovial fluid and plasma would change during progression of experimental osteoarthritis. In this study, we conducted small RNA sequencing to provide a comprehensive overview of the temporal expression profiles of small non-coding transcripts carried by extracellular vesicles derived from plasma and synovial fluid for the first time in a posttraumatic model of equine osteoarthritis. Additionally, we characterised synovial fluid and plasma-derived extracellular vesicles with respect to quantity, size, and surface markers. The different temporal expressions of seven microRNAs in plasma and synovial fluid-derived extracellular vesicles, eca-miR-451, eca-miR-25, eca-miR-215, eca-miR-92a, eca-miR-let-7c, eca-miR-486-5p, and eca-miR-23a, and four snoRNAs, U3, snord15, snord46, and snord58, represent potential biomarkers for early osteoarthritis. Bioinformatics analysis of the differentially expressed microRNAs in synovial fluid highlighted that in early osteoarthritis these related to the inhibition of cell cycle, cell cycle progression, DNA damage and cell proliferation as well as increased cell viability and differentiation of stem cells. Plasma and synovial fluid-derived extracellular vesicle small non-coding signatures have been established for the first time in a temporal model of osteoarthritis. These could serve as novel biomarkers for evaluation of osteoarthritis progression or act as potential therapeutic targets.

In contrast to traditional methods like real-time polymerase chain reaction, next-generation sequencing (NGS), and especially small RNA-seq, enables the untargeted investigation of the whole small RNAome, including microRNAs (miRNAs) but also a multitude of other RNA species. With the promising application of small RNAs as biofluid-based biomarkers, small RNA-seq is the method of choice for an initial discovery study. However, the presentation of specific quality aspects of small RNA-seq data varies significantly between laboratories and is lacking a common (minimal) standard. The miRNA NGS Discovery pipeline (miND) aims to bridge the gap between wet lab scientist and bioinformatics with an easy to setup configuration sheet and an automatically generated comprehensive report that contains all essential qualitative and quantitative results that should be reported. Besides the standard steps like preprocessing, mapping, visualization, and quantification of reads, the pipeline also incorporates differential expression analysis when given the appropriate information regarding sample groups. Although miND has a focus on miRNAs, other RNA species like tRNAs, piRNA, snRNA, or snoRNA are included and mapping statistics are available for further analysis. miND has been developed and tested on a multitude of data sets with various RNA sources (tissue, plasma, extracellular vesicles, urine, etc.) and different species. miND is a Snakemake based pipeline and thus incorporates all advantages using a flexible workflow management system. Reference databases are downloaded, prepared and built with an included (but separate) workflow and thus can easily be updated to the most recent version but also stored for reproducibility. In conclusion, the miND pipeline aims to streamline the bioinformatics processing of small RNA-seq data by standardizing the processing from raw data to a final, comprehensive and reproducible report.

BackgroundTo prevent or mitigate long-lasting learning problems and emotional, behavioral, and social-adaption difficulties associated with language disorders, age-appropriate German language competence at school entry level is essential. Therefore, universal screening of children in their penultimate year of pre-school has been established in Upper Austria. So far, the screenings administered by speech and language pathologists to identify risk of language disorder (LD) were not based on standardized materials.ObjectiveTo develop a screening instrument to identify increased risk of LD and to evaluate its validity and feasibility within the constraints of regular universal pre-school language screening.DesignA two-component screening instrument including direct assessment of expressive and receptive grammar was used in a sample of 374 children with German as their dominant language attending a public pre-school in their penultimate year (age 4-5 ½ years) in the state of Upper Austria. Assessment by use of standardized German language tests including a variety of linguistic domains was considered reference standard for diagnosing LD. Feasibility was assessed by a self-developed questionnaire completed by the administrators of the screening.ResultsThe combination of the expressive and receptive grammar scales demonstrated excellent accuracy (area under the curve score 0.928). A cut-off of 18 resulted in a failing rate of 21.8% and showed good sensitivity (84.2%) and specificity (85.3%). Acceptance by children and testers, time-economy and sustainability of the screening were mostly rated as high.

Abstract High electric fields that occur in thunderstorm clouds in the Earth’s atmosphere might accelerate energetic charged particles produced by cosmic rays. Such energetic particles, especially electrons, can cause additional ionization as they are multiplied and thus form avalanche of relativistic electrons. These relativistic electrons emit Bremsstrahlung in the X- or gamma-ray spectral ranges as they lose their kinetic energy via collisions. Thunderstorm ground enhancements (TGEs) of secondary cosmic ray fluxes recorded at the top of a sharp rocky mountain of Lomnický Štít in High Tatras (2634 m, Slovak Republic) are compared with simultaneous measurements of electric field at the mountain top and on its slope at the observatory of Skalnaté Pleso (1780 m). Results of measurements performed from May to September in 2017 and from May to October in 2018 are presented. The cosmic ray flux is measured by Space Environment Viewing and Analysis Network (SEVAN) and by neutron monitor with 1-s resolution. The TGEs that persisted usually several minutes were mainly detected in the SEVAN channel 1 which has the lowest energy threshold, about 7–8 MeV. A statistical analysis shows that these enhancements usually occurred (not only) during large values of vertical, upward-pointing electric fields measured just above the detector. It is shown that the measurement of electric field at Skalnaté Pleso, distant about 1.86 km from the mountain top is also partly correlated with the enhancements and can provide additional useful information about the distance or dimension of charge structure and dynamics of electric field, especially on short time scales. The enhancements usually did not exceed several tens of percent of background values. However, events that exceeded the background values several times were also recorded. The most extreme event exceeded the background values about 215 times. This event was also detected by other SEVAN channels and by the neutron monitor (~ 130% enhancement), which indicates a possibility of photonuclear reactions. The enhancements were often terminated by a nearby lightning.

Age‐induced decline in osteogenic potential of bone marrow mesenchymal stem cells (BMSCs) potentiates osteoporosis and increases the risk for bone fractures. Despite epidemiology studies reporting concurrent development of vascular and bone diseases in the elderly, the underlying mechanisms for the vascular‐bone cross‐talk in aging are largely unknown. In this study, we show that accelerated endothelial aging deteriorates bone tissue through paracrine repression of Wnt‐driven‐axis in BMSCs. Here, we utilize physiologically aged mice in conjunction with our transgenic endothelial progeria mouse model (Hutchinson‐Gilford progeria syndrome; HGPS) that displays hallmarks of an aged bone marrow vascular niche. We find bone defects associated with diminished BMSC osteogenic differentiation that implicate the existence of angiocrine factors with long‐term inhibitory effects. microRNA‐transcriptomics of HGPS patient plasma combined with aged‐vascular niche analyses in progeria mice reveal abundant secretion of Wnt‐repressive microRNA‐31‐5p. Moreover, we show that inhibition of microRNA‐31‐5p as well as selective Wnt‐activator CHIR99021 boosts the osteogenic potential of BMSCs through de‐repression and activation of the Wnt‐signaling, respectively. Our results demonstrate that the vascular niche significantly contributes to osteogenesis defects in aging and pave the ground for microRNA‐based therapies of bone loss in elderly. [ABSTRACT FROM AUTHOR]

Dysregulation of platelet function is causally connected to thrombus formation and cardiovascular diseases. Therefore, assessing platelet reactivity is crucial. However, current platelet function tests come with pitfalls, limiting clinical use. Plasma miRNA signatures have been suggested as novel biomarkers for predicting/diagnosing cardiovascular diseases and monitoring antiplatelet therapy. Here, we provide results from a comprehensive study on the feasibility of using circulatory platelet miRNAs as surrogate markers of platelet activation. We performed small RNA-Seq on different blood cell types to confirm known and identify novel platelet-enriched miRNAs and validated a panel of 16 miRNAs using RT-qPCR. To identify the main carrier of these blood-based platelet miRNAs, we enriched and analyzed distinct microvesicle populations. Platelets were stimulated with GPVI and P2Y12 agonists in vitro to monitor the release of the selected miRNAs following activation. Finally, the miRNA panel was also measured in plasma from mice undergoing the Folts intervention (recurrent thrombus formation in the carotid artery). Applying an unbiased bioinformatics-supported workflow to our NGS data, we were able to confirm a panel of previously established miRNA biomarker candidates and identify three new candidates (i.e., miR-199a-3p, miR-151a-5p, and miR-148b-3p). Basal levels of platelet-derived miRNAs in plasma were mainly complexed with proteins, not extracellular vesicles. We show that changes in miRNA levels due to platelet activation are detectable using RT-qPCR. In addition, we highlight limitations of studying the in vitro release of miRNAs from platelets. In vivo thrombosis resulted in significant elevations of platelet-derived miRNA levels in mice. In conclusion, we provide in-depth evidence that activated platelets release miRNAs, resulting in measurable changes in circulatory miRNA levels, rendering them promising biomarker candidates.

Lightning causes significant damage and casualties globally by directly striking humans and livestock, by igniting forest fires, and by inducing electrical surges in electronic infrastructure, airplanes, rockets, etc. Monitoring the evolution of thunderstorms by tracking lightning events using lightning locating systems can help prepare for and mitigate these disasters. In this work, we propose to use Benford’s law to assess the quality of the data provided by lightning locating systems. The Jensen–Shannon and Wasserstein distances between the recorded data distribution and Benford’s distribution are used as metrics for measuring the performance of the lightning locating systems. The data are provided by the European lightning detection network (EUCLID) for the years from 2000 to 2020. The two decades consist of three time windows between which the lightning locating system underwent several upgrades to improve the detection of both positive and negative strokes. The analysis shows that the agreement with Benford’s law is consistent with the expected behavior caused by the applied upgrades to the system throughout the years. The study suggests that the proposed approach can be used to test the success of software and hardware upgrades and to monitor the performance of lightning locating systems.