Confocal fluorescence microscopy and confocal raman microspectroscopy of X-ray irradiated LIF crystals

Radiation-induced color centers locally produced in lithium fluoride (LiF) are successfully used for radiation detectors. LiF detectors for extreme ultraviolet radiation, soft and hard X-rays, based on photoluminescence from aggregate electronic defects, are currently under development for imaging applications with laboratory radiation sources, as well as large-scale facilities. Among the peculiarities of LiF-based detectors, noteworthy ones are their very high intrinsic spatial resolution across a large field of view, wide dynamic range, and versatility. LiF crystals irradiated with a monochromatic 8 keV X-ray beam at KIT synchrotron light source (Karlsruhe, Germany) and with the broadband white beam spectrum of the synchrotron bending magnet have been investigated by optical spectroscopy, laser scanning confocal microscopy in fluorescence mode, and confocal Raman micro-spectroscopy. The 3D reconstruction of the distributions of the color centers induced by the X-rays has been performed with both confocal techniques. The combination of the LiF crystal capability to register volumetric X-ray mapping with the optical sectioning operations of the confocal techniques has allowed performing 3D reconstructions of the X-ray colored volumes and it could provide advanced tools for 3D X-ray detection

The GLP-1 receptor agonists exenatide and liraglutide activate Glucose transport by an AMPK-dependent mechanism

Additional file 2: Figure S2. Effects of EXE on Glut-4 in cultured L6 myotubes. Myotubes were stimulated with 100 nmol/l EXE for 20 min or 48 h. Panel A shows qPCR of Glut-4 mRNA. In panel B is a representative western blot for Glut-4 and β-Actin (loading control). In panel C is a representative western blot for Glut-4 and β-IR (loading control) in plasma membrane (PM) extracts (Glut-4 translocation). For A and C panels, data are shown as fold increase over control ± SD of three independent experiments (*p < 0.001, vs Ctrl)

CMS physics technical design report : Addendum on high density QCD with heavy ions

Peer reviewe

A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

The role of Methylglyoxal in the pathogenesis of endothelial insulin resistance and vascular damage in type 2 diabetes

It has now become evident that insulin exerts a direct action on vascular cells, thereby conditioning the outcome and progression of vascular complication associated with diabetes. However, the mechanisms through which insulin signaling is impaired in the vascular endothelium remain still unclear. Chronic hyperglycaemia per se promotes insulin resistance and plays a pivotal role in the outcome and progression of diabetes-associated vascular complications. Hyperglycaemia may act through different mechanisms, including generation of advanced glycation end products (AGEs). In this work we evaluated the role of the AGEs precursor methylglyoxal (MGO) in the generation of endothelial insulin-resistance in cellular and animal models. Time-courses experiments were performed on bovine aortic endothelial cells (BAEC) incubated with different concentrations of MGO. The glyoxalase 1 inhibitor “SpBrBzGSHCp2” was used to increase the endogenous levels of MGO. For the in vivo study, C57bl6 mice were intraperitoneally injected with a MGO solution at steadily increasing concentrations (50 to 75mg/kg) for 7 weeks. MGO incubation induces a 50% reduction of IRS1 phosphorylation, the loss of IRS1-p85 interaction and of the downstream Akt activation in response to insulin, whilst MAPK is more active in BAEC treated with MGO. The insulin-induced Akt activation is reverted by the inhibition of ERK through the use of MEK inhibitor U0126 in BAEC treated with MGO. Furthermore, downstream Akt, MGO is able to inhibit eNOS activation in response to insulin, and this was paralleled by a 60% decrease of insulin-induced NO production in BAEC. Similar results were obtained in BAEC treated with SpBrBzGSHCp2 compared to controls. Intraperitoneally administration of MGO to mice caused insulin resistance (ITT AUC: C57MGO 10163±1979 vs C57 7787±1174 mg/dl/120’, p=0.01) and reduced serum NO by 2.5-fold compared to untreated mice. Western blots of lysates of aortae from MG-treated mice revealed a reduction of insulin-induced Akt activation. In conclusion, this work shows that MGO impairs insulin signaling in endothelial cells and insulin effect on endothelial NO production both in vitro and in vivo. A possible role in these effects may be played by ERK. Further investigations of the molecular mechanisms by which hyperglycaemia compromises insulin action in vascular cells may allow to develop new strategies to preserve endothelial function in diabetic subjects

Methylglyoxal-Glyoxalase 1 Balance: The Root of Vascular Damage

The highly reactive dicarbonyl methylglyoxal (MGO) is mainly formed as byproduct of glycolysis. Therefore, high blood glucose levels determine increased MGO accumulation. Nonetheless, MGO levels are also increased as consequence of the ineffective action of its main detoxification pathway, the glyoxalase system, of which glyoxalase 1 (Glo1) is the rate-limiting enzyme. Indeed, a physiological decrease of Glo1 transcription and activity occurs not only in chronic hyperglycaemia but also with ageing, during which MGO accumulation occurs. MGO and its advanced glycated end products (AGEs) are associated with age-related diseases including diabetes, vascular dysfunction and neurodegeneration. Endothelial dysfunction is the first step in the initiation, progression and clinical outcome of vascular complications, such as retinopathy, nephropathy, impaired wound healing and macroangiopathy. Because of these considerations, studies have been centered on understanding the molecular basis of endothelial dysfunction in diabetes, unveiling a central role of MGO-Glo1 imbalance in the onset of vascular complications. This review focuses on the current understanding of MGO accumulation and Glo1 activity in diabetes, and their contribution on the impairment of endothelial function leading to diabetes-associated vascular damage

Dicarbonyl Stress at the Crossroads of Healthy and Unhealthy Aging

Dicarbonyl stress occurs when dicarbonyl metabolites (i.e., methylglyoxal, glyoxal and 3-deoxyglucosone) accumulate as a consequence of their increased production and/or decreased detoxification. This toxic condition has been associated with metabolic and age-related diseases, both of which are characterized by a pro-inflammatory and pro-oxidant state. Methylglyoxal (MGO) is the most reactive dicarbonyl and the one with the highest endogenous flux. It is the precursor of the major quantitative advanced glycated products (AGEs) in physiological systems, arginine-derived hydroimidazolones, which accumulate in aging and dysfunctional tissues. The aging process is characterized by a decline in the functional properties of cells, tissues and whole organs, starting from the perturbation of crucial cellular processes, including mitochondrial function, proteostasis and stress-scavenging systems. Increasing studies are corroborating the causal relationship between MGO-derived AGEs and age-related tissue dysfunction, unveiling a previously underestimated role of dicarbonyl stress in determining healthy or unhealthy aging. This review summarizes the latest evidence supporting a causal role of dicarbonyl stress in age-related diseases, including diabetes mellitus, cardiovascular disease and neurodegeneration