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Microglia are resident myeloid cells in the central nervous system (CNS) that control homeostasis and protect CNS from damage and infections. Microglia and peripheral myeloid cells accumulate and adapt tumor supporting roles in human glioblastomas that show prevalence in men. Cell heterogeneity and functional phenotypes of myeloid subpopulations in gliomas remain elusive. Here we show single-cell RNA sequencing (scRNA-seq) of CD11b+ myeloid cells in naïve and GL261 glioma-bearing mice that reveal distinct profiles of microglia, infiltrating monocytes/macrophages and CNS border-associated macrophages. We demonstrate an unforeseen molecular heterogeneity among myeloid cells in naïve and glioma-bearing brains, validate selected marker proteins and show distinct spatial distribution of identified subsets in experimental gliomas. We find higher expression of MHCII encoding genes in glioma-activated male microglia, which was corroborated in bulk and scRNA-seq data from human diffuse gliomas. Our data suggest that sex-specific gene expression in glioma-activated microglia may be relevant to the incidence and outcomes of glioma patients. Microglia and peripheral myeloid cells are critically involved in the immunopathology of glioblastoma. Here the authors present single-cell sequencing data that assesses the phenotypic composition of CD11b + myeloid cells from a murine model of glioblastoma and suggest enhanced MHCII transcription which they additionally report from human cases of glioblastoma. [ABSTRACT FROM AUTHOR]

High-grade gliomas (HGGs), the most common and aggressive primary brain tumors in adults, inevitably recur due to incomplete surgery or resistance to therapy. Intratumoral genomic and cellular heterogeneity of HGGs contributes to therapeutic resistance, recurrence, and poor clinical outcomes. Transcriptomic profiles of HGGs at recurrence have not been investigated in detail. Using targeted sequencing of cancer-related genes and transcriptomics, we identified single nucleotide variations, small insertions and deletions, copy number aberrations (CNAs), as well as gene expression changes and pathway deregulation in 16 pairs of primary and recurrent HGGs. Most of the somatic mutations identified in primary HGGs were not detected after relapse, suggesting a subclone substitution during the tumor progression. We found a novel frameshift insertion in the ZNF384 gene which may contribute to extracellular matrix remodeling. An inverse correlation of focal CNAs in EGFR and PTEN genes was detected. Transcriptomic analysis revealed downregulation of genes involved in messenger RNA splicing, cell cycle, and DNA repair, while genes related to interferon signaling and phosphatidylinositol (PI) metabolism are upregulated in secondary HGGs when compared to primary HGGs. In silico analysis of the tumor microenvironment identified M2 macrophages and immature dendritic cells as enriched in recurrent HGGs, suggesting a prominent immunosuppressive signature. Accumulation of those cells in recurrent HGGs was validated by immunostaining. Our findings point to a substantial transcriptomic deregulation and a pronounced infiltration of immature dendritic cells in recurrent HGG, which may impact the effectiveness of frontline immunotherapies in the GBM management. Key messages: Most of the somatic mutations identified in primary HGGs were not detected after relapse. Focal CNAs in EGFR and PTEN genes are inversely correlated in primary and recurrent HGGs. Transcriptomic changes and distinct immune-related signatures characterize HGG recurrence. Recurrent HGGs are characterized by a prominent infiltration of immature dendritic and M2 macrophages. [ABSTRACT FROM AUTHOR]

Background: Malignant tumours release factors, which attract myeloid cells and induce their polarisation to pro-invasive, immunosuppressive phenotypes. Brain-resident microglia and peripheral macrophages accumulate in the tumour microenvironment of glioblastoma (GBM) and induce immunosuppression fostering tumour progression. Macrophage colony stimulating factors (CSFs) control the recruitment of myeloid cells during peripheral cancer progression, but it is disputable, which CSFs drive their accumulation in gliomas.Methods: The expression of CSF2 (encoding granulocyte-macrophage colony stimulating factor) was determined in TCGA datasets and five human glioma cell lines. Effects of stable CSF2 knockdown in glioma cells or neutralising CSF2 or receptor CSF2Rα antibodies on glioma invasion were tested in vitro and in vivo.Results: CSF2 knockdown or blockade of its signalling reduced microglia-dependent glioma invasion in microglia-glioma co-cultures. CSF2-deficient human glioma cells encapsulated in cell-impermeable hollow fibres and transplanted to mouse brains, failed to attract microglia, but stimulated astrocyte recruitment. CSF2-depleted gliomas were smaller, attracted less microglia and macrophages, and provided survival benefit in tumour-bearing mice. Apoptotic microglia/macrophages were detected in CSF2-depleted tumours.Conclusions: CSF2 is overexpressed in a subset of mesenchymal GBMs in association with high immune gene expression. Tumour-derived CSF2 attracts, supports survival and induces pro-tumorigenic polarisation of microglia and macrophages. [ABSTRACT FROM AUTHOR]

A new optical authentication and security system using optical variable nano/micro-structures (OVNs) is presented. The proposed design features a passive authentication method using a simple optical system found in common fabrication facilities. The passive authentication is obtained by insertion of an OVN image directly on a processing layer or divided between multiple layers of the fabrication process. Authentic fabrication process is validated when the proper alignment (reconstructed image, for example) at the end of the fabrication is achieved. Simple proof-of-concept devices with the OVN-based authentication system are presented along with the optical images of the resulting authentication patterns. [ABSTRACT FROM AUTHOR]

In this paper, an innovative method for estimating the respiratory flow and efforts is proposed and evaluated in various postures and flow rates. Three micro electro-mechanical system accelerometers were mounted on the suprasternal notch, thorax and abdomen of subjects in supine, prone and lateral positions to record the upper airway acceleration and the movements of the chest and abdomen wall. The respiratory flow and efforts were estimated from the recorded acceleration signals by applying machine learning methods. To assess the agreement of the estimated signals with the well-established measurement methods, standard error of measurement (SEM) was calculated and $$\rho = 1-{\rm SEM}$$ was estimated for every condition. A significant agreement between the estimated and reference signals was found ( $$\rho = 0.83, 0.82$$ and 0.89 for the estimated flow, thorax and abdomen efforts respectively). Additionally, the agreement of the estimated and reference flows was assessed by calculating the ratio of time at the tidal peak inspiration flow to the inspiration time ( $$t_{\rm PTIF}/t_{\rm I}$$ ) and the ratio of time at the tidal peak expiration flow to the expiration time ( $$t_{\rm PTEF}/t_{\rm E}$$ ). Overall mean and standard deviation of absolute value of differences between $${t}_{{\rm PTIF}}/{t}_{{\rm I}}$$ and $${t}_{{\rm PTEF}}/{t}_{{\rm E}}$$ ratios calculated for every breathing cycle of reference and estimated flow were 0.0035 (0.06) and 0.051 (0.032), respectively. The presented results demonstrate the feasibility of using the upper-body acceleration as a simple solution for long-term monitoring of respiratory features. [ABSTRACT FROM AUTHOR]

Multispectral imaging is a highly desirable modality for material-based analysis in diverse areas such as food production and processing, satellite-based reconnaissance, and biomedical imaging. Here, we present nanofilter-based multispectral videography (nMSV) in the 700 to 950 nm range made possible by the tunable extraordinary-optical-transmission properties of 3D metallic nanostructures. Measurements made with nMSV during a bolus injection of an intravascular tracer in the ear of a piglet resulted in spectral videos of the microvasculature. Analysis of the multispectral videos generated contrast measurements representative of arterial pulsation, the distribution of microvascular transit times, as well as a separation of the venous and arterial signals arising from within the tissue. Therefore, nMSV is capable of acquiring serial multispectral images relevant to tissue hemodynamics, which may have application to the detection and identification of skin cancer. [ABSTRACT FROM AUTHOR]

Most neurological diseases are associated with chronic inflammation initiated by the activation of microglia, which produce cytotoxic and inflammatory factors. Signal transducers and activators of transcription (STATs) are potent regulators of gene expression but contribution of particular STAT to inflammatory gene expression and STAT-dependent transcriptional networks underlying brain inflammation need to be identified. In the present study, we investigated the genomic distribution of Stat binding sites and the role of Stats in the gene expression in lipopolysaccharide (LPS)-activated primary microglial cultures. Integration of chromatin immunoprecipitation-promoter microarray data and transcriptome data revealed novel Stat-target genes including Jmjd3, Ccl5, Ezr, Ifih1, Irf7, Uba7, and Pim1. While knockdown of individual Stat had little effect on the expression of tested genes, knockdown of both Stat1 and Stat3 inhibited the expression of Jmjd3 and inflammatory genes. Transcriptional regulation of Jmjd3 by Stat1 and Stat3 is a novel mechanism crucial for launching inflammatory responses in microglia. The effects of Jmjd3 on inflammatory gene expression were independent of its H3K27me3 demethylase activity. Forced expression of constitutively activated Stat1 and Stat3 induced the expression of Jmjd3, inflammation-related genes, and the production of pro-inflammatory cytokines as potently as lipopolysacharide. Gene set enrichment and gene function analysis revealed categories linked to the inflammatory response in LPS and Stat1C + Stat3C groups. We defined upstream pathways that activate STATs in response to LPS and demonstrated contribution of Tlr4 and Il-6 and interferon-γ signaling. Our findings define novel direct transcriptional targets of Stat1 and Stat3 and highlight their contribution to inflammatory gene expression. Key Message: [ABSTRACT FROM AUTHOR]

Signal transducer and activator of transcription 3 (STAT3) and nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) are transcription factors involved in cell survival, inflammation and metastasis. Constitutively activated STAT3 is found in many cancers, including melanoma. To study the crosstalk between STAT3 and NFκB signaling and its role in regulation of cancer cell survival, we used RNA interference (RNAi) to down-regulate STAT3 expression in human melanoma cells. RNAi strategies including double-stranded RNA, small interfering RNA (siRNA), short hairpin RNA (shRNA) and microRNA are widely used to knock down disease-causing genes in a targeted fashion. We found that shRNAs up-regulate non-specific NFκB activity, while siRNA directed against STAT3 specifically increase NFκB activity. The basal survival of melanoma cells is unaffected by STAT3 knockdown—likely due to activation of pro-survival NFκB signaling. Whereas, owing to off-target effects, plasmid-transcribed shRNA affects melanoma survival. Our data show that shRNA-mediated gene silencing induces non-specific or off-target effects that may influence cell functions. [ABSTRACT FROM AUTHOR]

We report a 3D plasmonic nanostructure having an extraordinary optical transmission due to localized surface plasmon (LSP) coupling between nanoholes and nanodisks. The nanostructure contains a free-standing gold nanohole array (NHA) film above a cavity and an array of nanodisks at the bottom of the cavity that is aligned with the NHA. For the device, the LSP-mediated resonance position was dependent on the hole and nanodisk diameter as well as the separation distance. Also, the effect of LSP coupling between each hole and corresponding nanodisk became negligible for cavities deeper than 200 nm as observed as a disappearance of the LSP resonance. The greatest LSP resonance transmission and the highest electric field intensity were observed for the structure with the shallowest cavity. In addition, the structure had high surface plasmon resonance sensitivity and may have potential for surface-enhanced Raman spectroscopy and optical trapping applications. [ABSTRACT FROM AUTHOR]

Neuroprotective and/or neuroregenerative activity of FK506, its derivatives, and to a lesser extent cyclosporin A (CsA) in animal models of neurodegenerative diseases of different etiology have been reported. Here, we verified a hypothesis that the most likely mechanism of their neuroprotective action is inhibition of the early steps of inflammatory activation of microglia by interference with mitogen-activated protein kinase (MAPK) signaling. The effect of immunosuppressants on lipopolysaccharide (LPS)-induced changes in morphology, proliferation, and motility of rat primary microglial cultures was evaluated. FK506 and CsA directly inhibited LPS-induced microglia activation and inflammatory responses. While both drugs efficiently reduced the expression of iNOS and the release of nitric oxide, only FK506 strongly inhibited the expression of Cox-2 and secretion of the mature form of IL-1β. FK506 strongly reduced LPS-induced activation of MAPK, and its downstream signaling crucial for inflammatory responses. Comparative analysis of global gene expression in rat ischemic brains and in LPS-stimulated microglial cultures revealed many genes and signaling pathways regulated in the same way in both systems. FK506 treatment blocked a majority of genes induced by an ischemic insult in the cortex, in particular inflammatory/innate immunity and apoptosis-related genes. Microglia-mediated inflammation is considered as one of the most important components of brain injury after trauma or stroke; thus, effective and multifaceted blockade of microglial activation by FK506 has clinical relevance and potential therapeutic implications. [ABSTRACT FROM AUTHOR]

The article presents a study that evaluated a novel integrated sensor system developed to simultaneously and synchronously measure and record cardiac vibrations and cardiac potentials from a single compact site on the chest. Topics covered include the advantages and disadvantages of using cardiac imaging technology, the architecture of the novel integrated sensor system, and the benefits of using the proposed integrated sensor system.

n improved Tungsten light source system for photovoltaic cell testing made from low-cost, commercially available materials is presented as an alternative to standard expensive testing equipment. In this work, spectral correction of the Tungsten light source is achieved by increasing the color temperature to ∼5200 K using inexpensive commercially available filters. Spectral measurements of the enhanced light source reveal that a better spectrum match towards the solar spectrum is achieved than what has been previously demonstrated. Specifically, the improved solar spectrum match is achieved by substantial filtering of the infrared range. The proposed setup is used to evaluate the performance of both silicon and organic based photovoltaic cells. [ABSTRACT FROM AUTHOR]

Smart wireless sensor systems that incorporate multiple sensors often cannot be implemented on a single chip. Advanced integration and assembly allows for a more complex conjugation and configuration of multiple system modules implemented under different technologies together in a small tiny package. In tiny sensor systems such as these, three common challenges seen across most platforms are: the difficult test access due to non-standard assembly and packaging, the testing of multiple heterogeneous sensor species, and the strict dimensional requirements limiting availability of any built-in hardware for testing. We discuss the method of testing by modules for testing a multilayer mechanically flexible wireless multisensor platform. A hierarchical test flow is presented for verifying the functionalities and assessing the performance of the various modules of the system. We also present an example of a design for testing feature, a built-in test point access bus that improves reliability for test point access, reduces the cost of testing and overall system bill-of-material, as well, increases test channel bandwidth allowing for full access to all critical subsystem nodes. Lastly, we provide examples of subsystem performance assessment and verification testing of selected sensor species on the multisensor platform as well as the system power consumption versus transmission range, to illustrate the usefulness of the test concepts, flow, and features introduced. [ABSTRACT FROM AUTHOR]

Activation of Akt signalling pathway is frequently found in glioma cells and may contribute to their resistance to undergo apoptosis in response to conventional therapies. We found that cyclosporin A (CsA) induces apoptosis of C6 glioma cells, which is associated with transcriptional activation of fasL. In the present paper, we investigated an involvement of Akt signalling in the regulation of FasL expression in CsA-induced apoptosis. We demonstrated that the level of active Akt decreases significantly after CsA treatment, which results in the decrease of Forkhead phosphorylation and its translocation to the nucleus. It correlated with an increase of binding to the Forkhead-responsive element FHRE from the FasL promoter, as demonstrated by gel-shift assays. Although treatment with LY294002, a specific inhibitor of PI3?K, decreased the phosphorylation of Akt and increased Fkhr translocation to the nucleus, these events were not sufficient to induce FasL expression and apoptosis of C6 glioma cells. Interference with Akt/Forkhead signalling by membrane-targeted Akt or removal of the FKHR-binding sites from the FasL promoter significantly abolished its activation. These results indicate that downregulation of Akt signalling and activation of Forkhead is a prerequisite for the induction of FasL promoter. It may be clinically important for pharmacological intervention in gliomas.Oncogene (2003) 22, 7617-7627. doi:10.1038/sj.onc.1207137 [ABSTRACT FROM AUTHOR]

The availability of faster electronic components allows the design of more effective and efficient test equipments. However in high-speed applications, the effect of interconnects between the tester and the device under test “DUT” introduces ringing, overshoot and timing delay problems. In this paper we present an output high speed buffer which helps to cancel the overshoot, undershoot, and ringing. The buffer which has a unity gain, presents a high output current and introduces small delay. It is able to drive the comparator of the tester through the transmission line with minimum distortion of the signal. Compared with other approaches, the use of this output buffer provides good improvement of the signal. This output buffer which is designed for the interface between tester and DUT can be considered for communication between high speed devices in printed circuits boards. The calibration procedure is explained in order to determine the delay introduced by the buffer and to measure low and high voltage levels of the digital output signal of the buffer. [ABSTRACT FROM AUTHOR]

Built-in current sensor (BICS) is known to enhance test accuracy, defect coverage of quiescent current (IDDQ) testing method in CMOS VLSI circuits. For new deep-submicron technologies, BICSs become essential for accurate and practical IDDQ testing. This paper presents a new BICS suitable for power dissipation measurement and IDDQ testing. Although the BICS presented in this paper is dedicated to submicron technologies that require reduced supply voltage, it can also be used for applications and technologies requiring normal supply voltage. The proposed BICS has been extended for on-line measurement of the power dissipation using only an additional capacitor. Power dissipation measurement is important for safety-critical applications and battery-powered systems. A simple self-test approach to verify the functionality and accuracy of BICSs has also been introduced. The proposed BICS has been implemented and tested using an N-well CMOS 1.2 μm technology. Practical results demonstrate that a very good measurement accuracy can be achieved. [ABSTRACT FROM AUTHOR]

The performance analysis of VLSI integrated circuits (ICs) with flat tools is slow and even sometimes impossible to complete. Some hierarchical tools have been developed to speed up the analysis of these large ICs. However, these hierarchical tools suffer from a poor interaction with the CAD database and poorly automatized operations. We introduce a general hierarchical framework for performance analysis to solve these problems. The circuit analysis is automatic under the proposed framework. Information that has been automatically abstracted in the hierarchy is kept in database properties along with the topological information. A limited software implementation of the framework, PREDICT, has also been developed to analyze the delay performance. Experimental results show that hierarchical analysis CPU time and memory requirements are low if heuristics are used during the abstraction process. [ABSTRACT FROM AUTHOR]

Built-in temperature sensors increase the system reliability by predicting eventual faults caused by excessive chip temperatures. In this paper, simple and efficient built-in temperature sensors for the on-line thermal monitoring of microelectronics structures are introduced. The proposed temperature sensors produce a signal oscillating at a frequency proportional to the temperature of the microelectronics structure and therefore they are compatible to the oscillation-test method. The oscillation-test method is a low-cost and robust test method for mixed-signal integrated circuits based on transforming the circuit under test (CUT) to an oscillator. This paper presents the design and detailed characteristics of the sensors proposed based on the CMOS 1.2 µm technology parameters of Mitel S.C.C. Extensive post-layout simulations show that the oscillation frequency is very sensitive to temperature variations. The sensors proposed require very small power dissipation and silicon area. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) contains rare glioma stem-like cells (GSCs) with capacities of self-renewal, multi-lineage differentiation, and resistance to conventional therapy. Drug-induced differentiation of GSCs is recognized as a promising approach of anti-glioma therapy. Accumulating evidence suggests that unique properties of stem cells depend on autophagy. Here we demonstrate that BIX01294, an inhibitor of a G9a histone methyltransferase (introducing H3K9me2 and H3K27me3 repressive marks) triggers autophagy in human glioma cells. Pharmacological or genetic inhibition of autophagy decreased LC3-II accumulation and GFP-LC3 punctation in BIX01294-treated cells. GSCs-enriched spheres originating from glioma cells and GBM patient-derived cultures express lower levels of autophagy related (ATG) genes than the parental glioma cell cultures. Typical differentiation inducers that upregulate neuronal and astrocytic markers in sphere cultures, increase the level of ATG mRNAs. G9a binds to the promoters of autophagy (LC3B, WIPI1) and differentiation-related (GFAP, TUBB3) genes in GSCs. Higher H3K4me3 (an activation mark) and lower H3K9me2 (the repressive mark) levels at the promoters of studied genes were detected in serum-differentiated cells than in sphere cultures. BIX01294 treatment upregulates the expression of autophagy and differentiation-related genes in GSCs. Pharmacological inhibition of autophagy decreases GFAP and TUBB3 expression in BIX01294-treated GSCs suggesting that BIX01294-induced differentiation of GSCs is autophagy-dependent. [ABSTRACT FROM AUTHOR]