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Purpose: The purpose of this study was to investigate if lesion detection using a single "All-in-One" (AIO) window was non-inferior to lesion detection on conventional window settings in thoracic oncology chest computed tomography (CT) examinations., Materials and Methods: In a retrospective study, 50 consecutive chest CT examinations of 50 patients (31 men, 19 women; mean age 64±10 [SD] years, range: 35-82 years) containing 417 lesions, were reviewed by 6 radiologists, subdivided into 2 groups of 3 radiologists each, with similar levels of expertise in each group (senior staff member, junior staff member and radiology resident). All examinations were reviewed in conventional or AIO window settings by one of the groups. A 'lesion' was defined as any abnormality seen on the chest CT examination, including both benign and malignant lesions, findings in chest and upper abdomen, and measurable and non-measurable disease. Lesions were listed as 'missed' when they were not seen by at least two out of three observers. F-tests were used to evaluate the significance of the variables of interest within a mixed model framework and kappa statistics to report interobserver agreement., Results: On a reader level, 54/417 lesions (12.9%) were not detected by the senior staff member reading the studies in conventional window settings and 45/417 (10.8%) by the senior staff member reading the AIO images. For the junior staff member and radiology resident this was respectively 55/417 (13.2%) and 67/417 (16.1%) for the conventional window settings and 43/417 (10.3%) and 61/417 (14.6%) for the AIO window. On a lesion level, 68/417 (16.3%) were defined as 'missed' lesions (lesions not detected by at least 2 readers): 21/68 (30.9%) on the AIO-window, 30/68 (44.1%) on conventional views and 17/68 (25.0%) on both views. The use of the AIO window did not result in an increase of missed lesions (P>0.99). Interobserver agreement in both groups was similar (P=0.46). Regarding lesions that were categorized as 'missed' on the AIO window or on conventional window settings, there was no effect of location (chest or upper abdomen) (P=0.35), window (P=0.97) and organ (P=0.98)., Conclusions: A single AIO-window is non-inferior to multiple conventional window settings for lesion detection on chest CT examinations in thoracic oncology patients., (Copyright © 2019 Société française de radiologie. Published by Elsevier Masson SAS. All rights reserved.)

Purpose: A newly developed image processing technique fuses conventional windows into a single 'All-In-One' (AIO) window. This study aims to evaluate variability of CT measurement of lesions in thoracic oncology patients on this novel AIO-window., Methods: Six radiologists with different levels of expertise measured 368 lesions of various size, origin and sharpness. All lesions were measured twice on the AIO-window and twice on the conventional window settings. Intraclass correlation coefficients and Bland-Altman plots were used to assess intra- and interobserver variability., Results: Overall intra-observer agreement for lesion diameters on the AIO-window and conventional window settings was 0.986 (95% Confidence interval (CI): 0.983-0.989) and 0.991 (95% CI 0.989-0.993) respectively. For interobserver agreement this was 0.982 (95% CI 0.979-0.985) (AIO) and 0.979 (95% CI 0.957-0.982) (conventional). For both the AIO and conventional windows, intra- and interobserver agreement were dependent on size, sharpness and reader experience. Measurement variability decreased with increasing lesion size. Regarding sharpness, inter- and intra-observer agreement ranged from 0.986-0.989 (AIO) and 0.985-0.992 (conventional) for well-defined lesions and from 0.978-0.983 (AIO) and 0.974-0.991 (conventional) for ill-defined lesions., Conclusions: Lesion diameters were consistently smaller on the AIO-window compared to conventional window settings. Overall intra- and interobserver variability rates were similar for the AIO-window and conventional window settings. We conclude that the AIO-window offers a reliable and reproducible alternative for measurement of thoracic lesions.

The establishment and validation of reliable induced pluripotent stem cell (iPSC)-derived in vitro models to study microglia and monocyte/macrophage immune function holds great potential for fundamental and translational neuro-immunology research. In this study, we first demonstrate that ramified CX 3 CR1 + iPSC-microglia (cultured within a neural environment) and round-shaped CX 3 CR1 - iPSC-macrophages can easily be differentiated from newly established murine CX 3 CR1 eGFP/+ CCR2 RFP/+ iPSC lines. Furthermore, we show that obtained murine iPSC-microglia and iPSC-macrophages are distinct cell populations, even though iPSC-macrophages may upregulate CX 3 CR1 expression when cultured within a neural environment. Next, we characterized the phenotypical and functional properties of murine iPSC-microglia and iPSC-macrophages following classical and alternative immune polarisation. While iPSC-macrophages could easily be triggered to adopt a classically-activated or alternatively-activated phenotype following, respectively, lipopolysaccharide + interferon γ or interleukin 13 (IL13) stimulation, iPSC-microglia and iPSC-macrophages cultured within a neural environment displayed a more moderate activation profile as characterised by the absence of MHCII expression upon classical immune polarisation and the absence of Ym1 expression upon alternative immune polarisation. Finally, extending our preceding in vivo studies, this striking phenotypical divergence was also observed for resident microglia and infiltrating monocytes within highly inflammatory cortical lesions in CX 3 CR1 eGFP/+ CCR2 RFP/+ mice subjected to middle cerebral arterial occlusion (MCAO) stroke and following IL13-mediated therapeutic intervention thereon. In conclusion, our study demonstrates that the applied murine iPSC-microglia and iPSC-macrophage culture models are able to recapitulate in vivo microglia and monocyte/macrophage ontogeny and corresponding phenotypical/functional properties upon classical and alternative immune polarisation, and therefore represent a valuable in vitro platform to further study and modulate microglia and (infiltrating) monocyte immune responses under neuro-inflammatory conditions within a neural environment., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Neuroglobin (Ngb) is a REST/NRSF-regulated protein, active in reactive oxygen species detoxification and cytochrome c inhibition, which provides a beneficial outcome in pathologies as Alzheimer's disease and strokes. Considering that oxidative stress and cell death are typical hallmarks of amyotrophic lateral sclerosis (ALS), we sought to explore Ngb's involvement along this disease progression. Ngb transcription was detected to be two-fold down-regulated in late-stage SOD G93A mice, similarly as previously described for Alzheimer disease. Interestingly, in accordance with REST/NRSF transcription, Ngb expression is higher in spinal cords than in cortices. Hence, downstream REST/NRSF mechanisms were studied. A methylation cluster in Ngb's exon 1 (Chr12:87101763-87102586) was selected to assess methylation alterations, based on significantly altered positions in GEO DataSets of human c9orf72 and sporadic ALS cases. However, only the methylation percentage on position Chr12.87102586 was significantly increased in SOD G93A mice. A larger impact can therefore be expected from the detected altered REST splicing; with levels of alternatively spliced, gene-activating REST4 to be lower than those of the gene-inhibitory full variant. To look further into the link between Ngb and ALS, we generated a double mutant Ngb -/- SOD G93A mouse model, which shows an earlier onset and severity of hind limb deficits. Mitochondria derived thereof showed an altered mean volume, granularity and Ca 2+ -induced swelling as compared to Ngb Wt/Wt SOD G93A mice. These results indicate Ngb to be involved in and affected by the SOD1 G93A pathology, which could in part be attributed to its role in halting destabilizing events of mitochondrial swelling and phenotypes., (Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.)

Neuroglobin (Ngb) is an evolutionary conserved member of the globin family with a primary expression in neurons of which the exact functions remain elusive. A plethora of in vivo and in vitro model systems has been generated to this day to determine the functional biological roles of Ngb. Here, we provide a comprehensive overview and discussion of the different Ngb models, covering animal and cellular models of both overexpression and knockout strategies. Intriguingly, an in-depth literature search of available Ngb expression models revealed crucial discrepancies in the outcomes observed in different models. Not only does the level of Ngb expression-either physiologically, overexpressed, or downregulated-alter its functional properties, the experimental setup, being in vitro or in vivo , does impact the functional outcome as well and, hence, whether or not a physiological and/or therapeutic role is ascribed to Ngb. These differences could highlight either technical or biological adaptations and should be considered until elucidation of the Ngb biology.

After its discovery in 2000, the notion grew that neuroglobin, a neuronal specific heme protein, is involved in cytoprotection. To date, neuroglobin levels have been positively correlated with a beneficial outcome in a plethora of neurotoxic insults, e.g., ischemic and traumatic brain injuries and Alzheimer's disease. The first part of this review goes further into these changes of neuroglobin expression upon different neuronal insults as well as the underlying regulation. In the second part, we shed light on the mechanisms by which neuroglobin contributes to neuroprotection, being (i) the scavenging and detoxification of reactive oxygen/nitrogen species, (ii) the augmentation of the threshold for apoptosis initiation, (iii) its contribution to an anti-inflammatory milieu, and (iv) tissue regeneration. We also consider different neuroglobin models to address as yet unanswered questions. Based on the recent findings and progress in the field, we invigorate the avenues of neuroglobin in neurological ailments to increase in the coming years.

In the quest to unravel its functional significance, neuroglobin (Ngb), a brain-specific neuroprotective protein, has recently been proposed as an actor in neurodevelopment. As neural stem cells (NSCs) are fundamental during brain development, the present study aimed at investigating the role of Ngb in the growth and proliferation of NSCs by comparing an Ngb-floxed (Ngb fl -)NSC line, equivalent to the wild-type cellular situation, with an in-house created Ngb knockout (Ngb KO -)NSC line. Ngb KO -NSCs were characterized by an increased growth and proliferation capacity in vitro, supported by RNA sequencing and western blot results reporting the downregulation of Cdkn1a and the upregulation of Cdk6, both enhancing the cell cycle. Based on additional gene ontology enrichment and pathway analyses, we hypothesize that the loss of Ngb affects multiple cellular signaling pathways with the most important being the Akt-Tp53 axis.

Neuroglobin (NGB) is an oxygen-binding protein that is mainly expressed in nervous tissues where it is considered to be neuroprotective during ischemic brain injury. Interestingly, transgenic mice overexpressing NGB reveal cytoprotective effects on tissues lacking endogenous NGB, which might indicate a therapeutic role for NGB in a broad range of ischemic conditions. In the present study, we investigated the effect of NGB overexpression on survival as well as on the size and occurrence of myocardial infarctions (MI) in a mouse model of acute MI (AMI) and a model of advanced atherosclerosis (ApoE -/- Fbn1 C1039G+/- mice), in which coronary plaques and MI develop in mice being fed a Western-type diet. Overexpression of NGB significantly enhanced post-AMI survival and reduced MI size by 14% 1 week after AMI. Gene expression analysis of the infarction border showed reduction of tissue hypoxia and attenuation of hypoxia-induced inflammatory pathways, which might be responsible for these beneficial effects. In contrast, NGB overexpression did not affect survival or occurrence of MI in the atherosclerotic mice although the incidence of coronary plaques was significantly reduced. In conclusion, NGB proved to act cytoprotectively during MI in the acute setting while this effect was less pronounced in the atherosclerosis model.

The true function of neuroglobin (Ngb) and, particularly, human Ngb (NGB) has been under debate since its discovery 15 years ago. It has been expected to play a role in oxygen binding/supply, but a variety of other functions have been put forward, including NO dioxygenase activity. However, in vitro studies that could unravel these potential roles have been hampered by the lack of an Ngb-specific reductase. In this work, we used electrochemical measurements to investigate the role of an intermittent internal disulfide bridge in determining NO oxidation kinetics at physiological NO concentrations. The use of a polarized electrode to efficiently interconvert the ferric (Fe(3+)) and ferrous (Fe(2+)) forms of an immobilized NGB showed that the disulfide bridge both defines the kinetics of NO dioxygenase activity and regulates appearance of the free ferrous deoxy-NGB, which is the redox active form of the protein in contrast to oxy-NGB. Our studies further identified a role for the distal histidine, interacting with the hexacoordinated iron atom of the heme, in oxidation kinetics. These findings may be relevant in vivo, for example, in blocking apoptosis by reduction of ferric cytochrome c, and gentle tuning of NO concentration in the tissues., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)

Transplantation of mesenchymal stem cells (MSCs) into injured or diseased tissue-for the in situ delivery of a wide variety of MSC-secreted therapeutic proteins-is an emerging approach for the modulation of the clinical course of several diseases and traumata. From an emergency point-of-view, allogeneic MSCs have numerous advantages over patient-specific autologous MSCs since "off-the-shelf" cell preparations could be readily available for instant therapeutic intervention following acute injury. Although we confirmed the in vitro immunomodulatory capacity of allogeneic MSCs on antigen-presenting cells with standard coculture experiments, allogeneic MSC grafts were irrevocably rejected by the host's immune system upon either intramuscular or intracerebral transplantation. In an attempt to modulate MSC allograft rejection in vivo, we transduced MSCs with an interleukin-13 (IL13)-expressing lentiviral vector. Our data clearly indicate that prolonged survival of IL13-expressing allogeneic MSC grafts in muscle tissue coincided with the induction of an alternatively activated macrophage phenotype in vivo and a reduced number of alloantigen-reactive IFNγ- and/or IL2-producing CD8(+) T cells compared to nonmodified allografts. Similarly, intracerebral IL13-expressing MSC allografts also exhibited prolonged survival and induction of an alternatively activated macrophage phenotype, although a peripheral T cell component was absent. In summary, this study demonstrates that both innate and adaptive immune responses are effectively modulated in vivo by locally secreted IL13, ultimately resulting in prolonged MSC allograft survival in both muscle and brain tissue. Stem Cells 2016;34:1971-1984., (© 2016 The Authors Stem Cells published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.)

Moderate levels of reactive oxygen species (ROS) are now recognized as redox signalling molecules. However, thus far, only mitochondria and NADPH oxidases have been identified as cellular sources of ROS in signalling. Here we identify a globin (GLB-12) that produces superoxide, a type of ROS, which serves as an essential signal for reproduction in C. elegans. We find that GLB-12 has an important role in the regulation of multiple aspects in germline development, including germ cell apoptosis. We further describe how GLB-12 displays specific molecular, biochemical and structural properties that allow this globin to act as a superoxide generator. In addition, both an intra- and extracellular superoxide dismutase act as key partners of GLB-12 to create a transmembrane redox signal. Our results show that a globin can function as a driving factor in redox signalling, and how this signal is regulated at the subcellular level by multiple control layers.