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The main purpose of the paper is to identify firm,- industry- and country-specific determinants of working capital management (WCM) in energy industry. The empirical research is based on 6122 EU companies operating in the years 2011–2018. The influence of internal factors on variables describing WCM (cash conversion cycle—CCC, financial liquidity—LIQ and level of working capital—WC) were identified. The factors included: size of the company (positive effect), its growth, tangibility and indebtedness (negative effect). Cash flow had a positive effect on CCC and a negative effect on LIQ and WC. The influence of industry-specific factors were also found. Companies applied similar strategies in CCC and LIQ management, following their industry averages. Measures of WCM decreased under the influence of an increase in average trade payables in the industry. Following country-specific factors were found to be significant: (i) growth of GDP and strength of legal rights had negative influence on all measures of WCM, (ii) unemployment positively affects LIQ and WC and negatively CCC, (iii) an increase in the share of renewable energy sources caused a decrease in all WCM measures, while (iv) with an increase in energy consumption, CCC and WC increased. [ABSTRACT FROM AUTHOR]

Highlights • A computational model of the respiratory control feedback loop exhibits realistic breath-to-breath variations in the amplitude and timing of breathing when random perturbations are applied to both the chemical and neural components of the circuit. • The largest alterations in the variability of breath amplitude and timing are caused by increases in lung resistance and impairments in gas exchange, both common features of pulmonary disease. • Breathing pattern variability may reflect discernible deterministic processes involved in the control of breathing, and may thus have diagnostic potential. Abstract The variability of the breath-to-breath breathing pattern, and its alterations in disease, may hold information of physiologic and/or diagnostic value. We hypothesized that this variability arises from the way that noise is processed within the respiratory feedback control loop, and that pathologic alterations to specific components within the system give rise to characteristic alterations in breathing pattern variability. We explored this hypothesis using a computational model of the respiratory control system that integrates mechanical factors, gas exchange processes, and chemoreceptor signals to simulate breathing patterns subject to the influences of random variability in each of the system components. We found that the greatest changes in the coefficient of variation (CV) of both breathing amplitude and timing were caused by increases in lung resistance and impairments in gas exchange, both common features of pulmonary disease. This suggests that breathing pattern variability may reflect discernible deterministic processes involved in the control of breathing. [ABSTRACT FROM AUTHOR]

In recent years the 3D printing methods have been developing rapidly. This article presents researches about a new composite consisted of golden and magnetite nanoparticles which could be used for this technique. Preparation of golden nanoparticles by laser ablation and their soldering by laser green light irradiation proceeded in water environment. Magnetite was obtained on chemical way. During experiments it was tested a change of a size of nanoparticles during laser irradiation, surface plasmon resonance, zeta potential. The obtained golden - magnetite composite material was magnetic after laser irradiation. On the end there was considered the application it for 3D printing devices, water filters and four-valued non-volatile memories. [ABSTRACT FROM AUTHOR]

The article presents a novel method of producing Gd particles and preserving them from oxidation. The particles were produced in liquid paraffin by means of AC electric discharge and stored in the solidified paraffin. After seven months, the surface of the Gd was found to be exempt of oxidation. Moreover a composite material formed from mixing paraffin with Gd particles was conductive and magnetic and also presented photovoltaic effect. This method is a promising means of producing, at an industrial scale, particles from materials extremely sensitive to environment such as rare earth materials. Also the new material consisted of Gd particles in a paraffin matrix can find applications in many branches of industry. [ABSTRACT FROM AUTHOR]

This paper presents a model of alveolar-capillary oxygen diffusion with dynamics of air transport through the respiratory tract. For this purpose electrical model representing the respiratory tract mechanics and differential equations representing oxygen membrane diffusion are combined. Relevant thermodynamic relations describing the mass of oxygen transported into the human body are proposed as the connection between these models, as well as the influence of ventilation-perfusion mismatch on the oxygen diffusion. The model is verified based on simulation results of varying exercise intensities and statistical calculations of the results obtained during various clinical trials. The benefit of the approach proposed is its application in simulation-based research aimed to generate quantitative data of normal and pathological conditions. Based on the model presented, taking into account many essential physiological processes and air transport dynamics, comprehensive and combined studies of the respiratory efficiency can be performed. The impact of physical exercise, precise changes in respiratory tract mechanics and alterations in breathing pattern can be analyzed together with the impact of various changes in alveolar-capillary oxygen diffusion. This may be useful in simulation of effects of many severe medical conditions and increased activity level. [ABSTRACT FROM AUTHOR]

This paper presents a new versatile approach to model severe human respiratory diseases via computer simulation. The proposed approach enables one to predict the time histories of various diseases via information accessible in medical publications. This knowledge is useful to bioengineers involved in the design and construction of medical devices that are employed for monitoring of respiratory condition. The approach provides the data that are crucial for testing diagnostic systems. This can be achieved without the necessity of probing the physiological details of the respiratory system as well as without identification of parameters that are based on measurement data. [ABSTRACT FROM AUTHOR]

Introduction. Tau protein is a potential marker of neuronal damage. The aim of the study is to investigate its potential role as a marker of brain atrophy in multiple sclerosis (MS). Materials and methods. Cerebrospinal fluid (CSF) and blood samples were collected from 48 patients with multiple sclerosis. Total-tau (t-tau) and phospho181Thr-tau (p-tau) concentrations were assayed with commercially available INNOTEST® hTAU Ag and INNOTEST® phospho181Thr-tau(181P) and correlated with indices of brain atrophy in magnetic resonance imaging (MRI) and clinical characteristics of the study population. Results. T-tau concentration in CSF was significantly higher in relapsing-remitting (RR) compared to secondary progressive (SP) MS patients ( P = 0.01). Brain parenchymal fraction (BPF) was significantly decreased in SP patients ( P = 0.002). BPF in the whole study population correlated inversely with Expanded Disability Status Scale (EDSS) ( r = -0.51, P = 0.0002) and Multiple Sclerosis Severity Score (MSSS) ( r = -0.42, P = 0.002). T-tau in CSF in the whole patient group correlated inversely with EDSS ( r = -0.58, P = 0.0006). Conclusions. The results of our study suggest that total-tau concentration in CSF in a MS population decreases in the course of disease and reflects degree of parenchymal brain loss. [ABSTRACT FROM AUTHOR]

In search of biological marker in multiple sclerosis (MS), total-tau and phospho-tau (Thr181) concentrations were established in CSF and serum of 78 patients with MS, using commercially available kits. Serum and CSF concentrations of IgG, IgM, and albumin were assayed simultaneously to calculate quotients and indices of intrathecal synthesis. Serum t-tau detection was strikingly low (23.1%); therefore, this factor was excluded from further analysis. Serum p-tau levels did not correlate with any of indices or quotients. Unexpectedly, CSF t-tau and p-tau showed an inverse relation with MSSS and EDSS, which has not been published elsewhere. Our results do not support utility of serum t-tau and p-tau as surrogate markers for MS. [ABSTRACT FROM AUTHOR]

Abstract: The present paper reports results on the Gd-doped Co–Cu granular alloys and metallic glasses prepared by different processing methods. The main aim of this research was to produce a ternary alloy which radically changes its magnetoresistance (MR) properties below critical temperature. A detailed study on the evolution of the structure and phase separation is presented basing on X-ray diffractometry (XRD) analysis and scanning electron microscopy (SEM) observation. MR was measured by means of a two point probe method in magnetic field using a new experimental device which was patented thereafter. [Copyright &y& Elsevier]

An essential prerequisite for safe transport and use of natural gas is their appropriate odorization. This enables the detection of uncontrolled gas leaks. Proper and systematic odorization inspection ensures both safe use of gas and continuity of the process itself. In practice, it is conducted through, among others, measuring odorant concentrations in gas. Control devices for rapid gas odorization measurements that are currently used on a large scale in the gas industry are equipped with electrochemical detectors selective for sulfur compounds like tetrahydrothiophene (THT). Because the selectivity of electrochemical detector response to one compound (e.g., THT), the available declarations of manufacturers show that detector sensitivity (indirectly also the quality of the measurement result) is influenced by the presence of increased e.g., sulfur or hydrogen compound content in the gas. Because of the lack of sufficient source literature data in this field, it was necessary to experimentally verify this impact. The results of studies on experimental verification of suspected influence of increased amounts of hydrogen in gas on the response of electrochemical detector was carried out at the Oil and Gas Institute—National Research Institute (INiG—PIB). They are presented in this article. The data gathered in the course of researching the dependence between THT concentration measurement result quality and hydrogen content in gas composition enabled a preliminary assessment of the threat to the safety of end users of gaseous fuels caused by the introduction of this gas into the distribution network. Noticing the scope of necessary changes in the area of odorization is necessary to guarantee this safety. [ABSTRACT FROM AUTHOR]

Summary: Dendritic spines are the major sites of excitatory synaptic input, and their morphological changes have been linked to learning and memory processes. Here, we report that growing microtubule plus ends decorated by the microtubule tip-tracking protein EB3 enter spines and can modulate spine morphology. We describe p140Cap/SNIP, a regulator of Src tyrosine kinase, as an EB3 interacting partner that is predominately localized to spines and enriched in the postsynaptic density. Inhibition of microtubule dynamics, or knockdown of either EB3 or p140Cap, modulates spine shape via regulation of the actin cytoskeleton. Fluorescence recovery after photobleaching revealed that EB3-binding is required for p140Cap accumulation within spines. In addition, we found that p140Cap interacts with Src substrate and F-actin-binding protein cortactin. We propose that EB3-labeled growing microtubule ends regulate the localization of p140Cap, control cortactin function, and modulate actin dynamics within dendritic spines, thus linking dynamic microtubules to spine changes and synaptic plasticity. [Copyright &y& Elsevier]

Abstract: Differentiated mammalian cells are often characterized by highly specialized and polarized structure. Its formation and maintenance depends on cytoskeletal components, among which microtubules play an important role. The shape and dynamic properties of microtubule networks are controlled by multiple microtubule-associated factors. These include molecular motors and non-motor proteins, some of which accumulate specifically at the growing microtubule plus-ends (the so-called microtubule plus-end tracking proteins). Plus-end tracking proteins can contribute to the regulation of microtubule dynamics, mediate the cross-talk between microtubule ends, the actin cytoskeleton and the cell cortex, and participate in transport and positioning of structural and regulatory factors and membrane organelles. Malfunction of these proteins results in various human diseases including some forms of cancer, neurodevelopmental disorders and mental retardation. In this article we discuss recent data on microtubule dynamics and activities of microtubule plus-end binding proteins important for the physiology and pathology of differentiated mammalian cells such as neurons, polarized epithelia, muscle and sperm cells. [Copyright &y& Elsevier]

Abstract: Actin cytoskeleton dynamics and membrane trafficking are tightly connected and are among the most important driving forces of neuronal development, basic synaptic transmission events, and synaptic plasticity. One group of proteins involved in coordination of these two processes is the family of ADP ribosylation factors (ARFs) regulating actin dynamics, lipid modification and membrane trafficking. ARF6 is the only member of the ARF family that can simultaneously regulate actin cytoskeleton changes and membrane exchange between plasma membrane and endocytic compartments. The presence of ARF6 and its guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) in the brain, as well as its capability to regulate several aspects of neuronal development and synaptic plasticity, has been recently demonstrated. The main purpose of this review is to present the current knowledge about how ARF6 can influence morphological processes crucial for proper formation of the neuronal circuits in the brain, including dendrite and axon differentiation, development of dendritic arbor complexity and dendritic spine formation. Potential effects of ARF6 on synaptic events resulting from its ability to control exo- and endocytosis will be also discussed [Copyright &y& Elsevier]

The syntheses and photophysical characterization of ZP9, 2-{2-chloro-6-hydroxy-3-oxo-5-[(2-{[pyridin-2-ylmethyl-(1H-pyrrol-2-ylmethyl)amino]methyl}phenylamino)methyl]-3H-xanthen-9-yl}benzoic acid, and ZP10, 2-{2-chloro-6-hydroxy-5-[(2-{[(1-methyl-1H-pyrrol-2-ylmethyl)pyridin-2-ylmethylamino]methyl}phenylamino)methyl]-3- oxo-3H-xanthen-9-yI}benzoic acid, two asymmetrically derivatized fluorescein-based dyes, are described. These sensors each contain an aniline-based ligand moiety functionalized with a pyridyl-amine-pyrrole group and have dissociation constants for Zn(II) in the sub-micromolar (ZP9) and low-micromolar (ZP10) range, which we define as "midrange". They give ~12- (ZP9) and ~7-fold (ZP10) fluorescence turn-on immediately following Zn(II) addition at neutral pH and exhibit improved selectivity for Zn(II) compared to the di-(2-picolyl)amine-based Zinpyr (ZP) sensors. Confocal microscopy studies indicate that such asymmetrical fluorescein-based probes are cell permeable and Zn(lI) responsive in vivo. [ABSTRACT FROM AUTHOR]

QZ1, 2-[2-chloro-6-hydroxy-3-oxo-5-(quinolin-8-ylaminomethyl)-3H-xanthen-9-yl]benzoic acid, and QZ2, 2-[6-hydroxy-3-oxo-4,5-bis-(quinolin-8-ylaminomethyl)-3H-xanthen-9-yl]benzoic acid, two fluorescein-based dyes derivatized with 8-aminoquinoline, have been prepared and their photophysical, thermodynamic, and zinc-binding kinetic properties determined. Because of their low background fluorescence and highly emissive Zn(II) complexes, QZ1 and QZ2 have a large dynamic range, with ∼42- and ∼150-fold fluorescence enhancements upon Zn(II) coordination, respectively. These dyes have micromolar Kd values for Zn(II) and are selective for Zn(II) over biologically relevant concentrations of the alkali and alkaline earth metals. The Zn(II) complexes also fluoresce brightly in the presence of excess Mn(II), Fe(II), Co(II), Cd(II), and Hg(II), offering improved specificity for Zn(II) over di(2-picolyl)amine-based Zn(II) sensors. Stopped-flow kinetic investigations indicate that QZ1 and QZ2 bind Zn(II) with kon values of (3-4) × 106 M-1 s-1, compared to (6-8) × 105 M-1 s-1 for select ZP (Zinpyr) dyes, at 4.3 °C. Dissociation of Zn(II) from QZ1 and QZ2 occurs with koff values of 150 and 160 s-1, over 5 orders of magnitude larger than those for ZP probes, achieving reversibility on the biological (millisecond) time scale. Laser scanning confocal and two-photon microscopy studies reveal that QZ2 is cell-permeable and Zn(II)-responsive in vivo. Because of its weaker affinity for Zn(II), QZ2 responds to higher concentrations of intracellular Zn(II) than members of the ZP family, illustrating that binding affinity is an important parameter for Zn(II) detection in vivo. [ABSTRACT FROM AUTHOR]

The molecular mechanisms that determine the size and complexity of the neuronal dendritic tree are unclear. Here, we show that the phosphoinositide-3′ kinase (PI3K)-Akt-mammalian target of rapamycin (mTOR) signaling pathway promotes the growth and branching of dendrites in cultured hippocampal neurons. Constitutively active mutants of Ras, PI3K, and Akt, or RNA interference (RNAi) knockdown of lipid phosphatase PTEN (phosphatase and tensin homolog deleted on chromosome Ten), induced growth and elaboration of dendrites that was blocked by mTOR inhibitor rapamycin and/or by overexpression of eIF-4E binding protein 1 (4E-BP1), which inhibits translation of 5′ capped mRNAs. The effect of PI3K on dendrites was lost in more mature neurons (> 14 d in vitro). Dendritic complexity was reduced by inhibition of PI3K and by RNAi knockdown of mTOR or p70 ribosomal S6 kinase (p70S6K, an effector of mTOR). A rapamycin-resistant mutant of mTOR "rescued" the morphogenetic effects of PI3K in the presence of rapamycin. By regulating global and/or local protein translation, and as a convergence point for multiple signaling pathways, mTOR could play a central role in the control of dendrite growth and branching during development and in response to activity. [ABSTRACT FROM AUTHOR]

An understanding of the microscopic details of growth processes is of key importance in materials science and technological applications. Experimentally, the most direct methods to study surface topography are real-space imaging techniques, such as, for example, scanning force microscopy. Co/Cu multilayers deposited on Si(1 0 0) precovered with various thicknesses of In buffer has been studied by scanning force microscopy. From the observed film morphologies we determined the evolution of a multilayer surface, indicating a complex growth process. [Copyright &y& Elsevier]

Zinc is an essential metal ion for human growth and development, the disruption of cellular Zn2+ homeostasis being implicated in several major disorders including Alzheimer's disease, diabetes, and cancer. The molecular mechanisms of Zn2+ physiology and pathology are insufficiently understood, however, owing in part to the lack of tools for measuring changes in intracellular Zn2+ concentrations with high spatial and temporal fidelity. To address this critical need, we have synthesized, characterized, and applied an intracellular fluorescent probe for the ratiometric imaging of Zn2+ based on a tautomeric seminaphthofluorescein platform. Zin-naphthopyr 1 (ZNP1) affords single-excitation, dual-emission ratiometric detection of intracellular Zn2+ through Zn2+-controlled switching between fluorescein and naphthofluorescein tautomeric forms. The probe features visible excitation and emission profiles, excellent selectivity responses for Zn2+ over competing Ca2+ and Mg2+ ions at intracellular concentrations, a dissociation constant (Kd) for Zn2+ of <1 nM, and an 18-fold increase in fluorescence emission intensity ratio (&lamda;624/&lamda;523) upon zinc binding. We demonstrate the value of the ZNP1 platform for biological applications by imaging changes in intracellular [Zn2+] in living mammalian cells. Included is the ratiometric detection of endogenous pools of intracellular Zn2+ after NO-induced release of Zn2+ from cellular metalloproteins. We anticipate that ZNP1 and related probes should find utility for interrogating the biology of Zn2+. [ABSTRACT FROM AUTHOR]

The acronym ICER (inducible cAMP early repressor) refers to a group of four proteins produced from the CREM/ICER gene due to use of an internal promoter (P2) placed in an intron of the CREM (cAMP responsive element modulator) gene. The ICER proteins contain DNA binding/leucine zipper domains that make them endogenous inhibitors of transcription driven by CREB (cAMP responsive element binding protein) and its cognates, CREM and ATF-1 (activating transcription factor-1). ICER expression is inducible in the brain and in neuronal culture by a variety of stimuli. As a CREB antagonist, ICER appears to be of pivotal importance in neuronal plasticity and programmed cell death. [ABSTRACT FROM AUTHOR]

Presents a study that investigated the role played by inducible cAMP early repressor in neuronal apoptosis. Materials and methods; Results; Discussion.

The aim of the study is to identify the main determinants of the capital structure of energy industry companies in the European Union. The study was based on a panel of 6122 companies from 25 EU countries, operating between 2011 and 2018. The study used multiple regression analysis. We have obtained strong evidence for a positive relationship between corporate debt and tangibility and size, and a negative relationship for profitability and liquidity. The factors that also affect the share of debt in capital have turned out to be growth (positive relationship) and non-debt tax shield (negative relationship), but the statistical significance of these relationships is ambiguous. We have shown that growth of industry business risk is accompanied by an increase in corporate debt and this is a distinguishing feature of the energy industry. For country-specific capital structure determinants, we have obtained strong evidence for the negative relationship between GDP growth, the level of stakeholder rights protection, the degree of capital markets development, and indebtedness of the companies studied. There has been moderate support for the hypotheses of a positive effect of inflation, taxation, and the degree of financial institutions development. Our study has also shown a negative impact of the volume of energy consumption and the share of renewable sources in its production and a positive impact of market monopolization on the indebtedness of companies from the energy industry in the EU. [ABSTRACT FROM AUTHOR]

One of the most important requirements concerning the quality of natural gases, guaranteeing their safe use, involves providing the proper level of their odorization. This allows for the detection of uncontrolled leakages of gases from gas networks, installations and devices. The concentration of an odorant should be adjusted in such a manner that the gas odor in a mixture with air would be noticeable by users (gas receivers). A permanent odor of gas is guaranteed by the stability of the odorant molecule and its resistance to changes in the composition of odorized gases. The article presents the results of experimental research on the impact of a hydrogen additive on the stability of tetrahydrothiophene (THT) mixtures in methane and in natural gas with a hydrogen additive. The objective of the work was to determine the readiness of measurement infrastructures routinely used in monitoring the process of odorizing natural gas for potential changes in its composition. One of the elements of this infrastructure includes the reference mixtures of THT, used to verify the correctness of the readings of measurement devices. The performed experimental tests address possible changes in the composition of gases supplied via a distribution network, resulting from the introduction of hydrogen. The lack of interaction between hydrogen and THT has been verified indirectly by assessing the stability of its mixtures with methane and natural gas containing hydrogen. The results of the presented tests permitted the identification of potential hazards for the safe use of gas from a distribution network, resulting from changes in its composition caused by the addition of hydrogen. [ABSTRACT FROM AUTHOR]

Abstract: Gold and magnetite (Fe3O4) composite nanoparticles were prepared by a combination of unique laser processes in water. Pulsed laser ablation in liquid (PLAL) was employed to produce high pure gold nanoparticles and the successful gold/magnetite composite nanoparticles formation by a ‘nano-soldering’ effect was confirmed. The irradiation effect on the magnetic properties of the magnetite nanoparticles was investigated. [Copyright &y& Elsevier]

Thermal gas meters represent a promising technology for billing customers for gaseous fuels, however, it is essential to ensure that measurement accuracy is maintained in the long term and in a broad range of operating conditions. The effect of hydrogen addition to natural gas will change the physicochemical properties of the mixture of natural gas and hydrogen. Such a mixture will be supplied through the gas system, to consumers, including households, where the amounts of received gas will be metered. The physicochemical properties of hydrogen, including the specific density or viscosity, differ significantly from those of the natural gas components, such as methane, ethane, propane, nitrogen, etc. Therefore, it is of utmost importance to establish the impact of the changes in the gas composition caused by the addition of hydrogen to natural gas on the metrological properties of household gas meters, including thermal gas meters. Furthermore, since household gas meters can be installed outdoors and, taking into account the fact that household gas meters are good heat exchangers, the influence of ambient and gas temperature on the metrological properties of those meters should be investigated. This article reviews a test bench and a testing method concerning errors of thermal gas meter indicators using air and natural gas, including the type containing hydrogen. The indication errors for thermal gas meters using air, natural gas and natural gas with an addition of 2%, 4%, 5%, 10% and 15% hydrogen were determined and then subjected to metrological analysis. Moreover, the test method and test bench are discussed and the results of tests on the impact of ambient and gas temperatures (‒25 °C and 55 °C, respectively) on the errors of indications of thermal gas meters are presented. Conclusions for distribution system operators in terms of gas meter selection were drawn based on the test results. [ABSTRACT FROM AUTHOR]

Power-to-gas technology plays a key role in the success of the energy transformation. This paper addresses issues related to the legal and technical regulations specifying the rules for adding hydrogen to the natural gas network. The main issue reviewed is the effects of the addition of hydrogen to natural gas on the durability of diaphragm gas meters. The possibility of adding hydrogen to the gas network requires confirmation of whether, within the expected hydrogen concentrations, long-term operation of gas meters will be ensured without compromising their metrological properties and operational safety. Methods for testing the durability of gas meters applied at test benches and sample results of durability tests of gas meters are presented. Based on these results, a metrological and statistical analysis was carried out to establish whether the addition of hydrogen affects the durability of gas meters over time. The most important conclusion resulting from the conducted study indicates that, for the tested gas meter specimens, there was no significant metrological difference between the obtained changes of errors of indications after testing the durability of gas meters with varying hydrogen content (from 0% to 15%). [ABSTRACT FROM AUTHOR]

Gold and magnetite composite nanoparticles have been prepared by a unique laser process. Both source colloid solutions of gold and magnetite were mixed and irradiated by an Nd:YAG laser (532 nm). The composite formation by a ‘nanosoldering’ effect was confirmed by microscope observations. [ABSTRACT FROM AUTHOR]

The endocytic adaptor protein 2 (AP-2) complex binds dynactin as part of its noncanonical function, which is necessary for dynein-driven autophagosome transport along microtubules in neuronal axons. The absence of this AP-2-dependent transport causes neuronal morphology simplification and neurodegeneration. The mechanisms that lead to formation of the AP-2-dynactin complex have not been studied to date. However, the inhibition of mammalian/mechanistic target of rapamycin complex 1 (mTORC1) enhances the transport of newly formed autophagosomes by influencing the biogenesis and protein interactions of Rab-interacting lysosomal protein (RILP), another dynein cargo adaptor. We tested effects of mTORC1 inhibition on interactions between the AP-2 and dynactin complexes, with a focus on their two essential subunits, AP-2β and p150Glued. We found that the mTORC1 inhibitor rapamycin enhanced p150Glued–AP-2β complex formation in both neurons and non-neuronal cells. Additional analysis revealed that the p150Glued–AP-2β interaction was indirect and required integrity of the dynactin complex. In non-neuronal cells rapamycin-driven enhancement of the p150Glued–AP-2β interaction also required the presence of cytoplasmic linker protein 170 (CLIP-170), the activation of autophagy, and an undisturbed endolysosomal system. The rapamycin-dependent p150Glued–AP-2β interaction occurred on lysosomal-associated membrane protein 1 (Lamp-1)-positive organelles but without the need for autolysosome formation. Rapamycin treatment also increased the acidification and number of acidic organelles and increased speed of the long-distance retrograde movement of Lamp-1-positive organelles. Altogether, our results indicate that autophagy regulates the p150Glued–AP-2β interaction, possibly to coordinate sufficient motor-adaptor complex availability for effective lysosome transport. [ABSTRACT FROM AUTHOR]

Introduction: Tuberous sclerosis complex (TSC) is caused by variants in TSC1/TSC2, leading to constitutive activation of the mammalian target of rapamycin (mTOR) complex 1. Therapy with everolimus has been approved for TSC, but variations in success are frequent. Recently, caudal late interneuron progenitor (CLIP) cells were identified as a common origin of the TSC brain pathologies such as subependymal giant cell astrocytomas (SEGA) and cortical tubers (CT). Further, targeting the epidermal growth factor receptor (EGFR) with afatinib, which is expressed in CLIP cells, reduces cell growth in cerebral TSC organoids. However, investigation of clinical patient‐derived data is lacking. Aims: Observation of EGFR expression in SEGA, CT and focal cortical dysplasia (FCD) 2B human brain specimen and investigation of whether its inhibition could be a potential therapeutic intervention for these patients. Methods: Brain specimens of 23 SEGAs, 6 CTs, 20 FCD2Bs and 17 controls were analysed via immunohistochemistry to characterise EGFR expression, cell proliferation (via Mib1) and mTOR signalling. In a cell‐based assay using primary patient‐derived cells (CT n = 1, FCD2B n = 1 and SEGA n = 4), the effects of afatinib and everolimus on cell proliferation and cell viability were observed. Results: EGFR overexpression was observed in histological sections of SEGA, CT and FCD2B patients. Both everolimus and afatinib decreased the proliferation and viability in primary SEGA, tuber and FCD2B cells. Conclusion: Our study demonstrates that EGFR suppression might be an effective alternative treatment option for SEGAs and tubers, as well as other mTOR‐associated malformations of cortical development, including FCD2B. [ABSTRACT FROM AUTHOR]

Static flow sensors (e.g., thermal gas micro electro-mechanical sensors—MEMS—and ultrasonic time of flight) are becoming the prevailing technology for domestic gas metering and billing since they show advantages in respect to the traditional volumetric ones. However, they are expected to be influenced in-service by changes in gas composition, which in the future could be more frequent due to the spread of hydrogen admixtures in gas networks. In this paper, the authors present the results of an experimental campaign aimed at analyzing the in-service reliability of both static and volumetric gas meters with different hydrogen admixtures. The results show that the accuracy of volumetric and ultrasonic meters is always within the admitted limits for subsequent verification and even within those narrower of the initial verification. On the other hand, the accuracy of the first generation of thermal mass gas flow sensors is within the limits of the verification only when the hydrogen admixture is below 2%vol. At higher hydrogen content, in fact, the absolute weighted mean error ranges between 3.5% (with 5%vol of hydrogen) and 15.8% (with 10%vol of hydrogen). [ABSTRACT FROM AUTHOR]

This article focuses on the selective labeling of extracellular proteins, which contains polyhistidine sequences by a fluorescein-nitrilotriacetic acid conjugate. Using fluorescent small molecules as the label can mitigate difficulties. Synthetic probes are generally less demanding sterically than proteins and they can be attached to the target peptide with minimal disruption of native function and structure. Photophysical and acid base properties can be modified more easily and changed to suit an application of interest. The set of methodologies which are used to attach the probes to biological targets is limited.

In neurons, the polarized distribution of vesicles and other cellular materials is established through molecular motors that steer selective transport between axons and dendrites. It is currently unclear whether interactions between kinesin motors and microtubule-binding proteins can steer polarized transport. By screening all 45 kinesin family members, we systematically addressed which kinesin motors can translocate cargo in living cells and drive polarized transport in hippocampal neurons. While the majority of kinesin motors transport cargo selectively into axons, we identified five members of the kinesin-3 ( KIF1) and kinesin-4 ( KIF21) subfamily that can also target dendrites. We found that microtubule-binding protein doublecortin-like kinase 1 ( DCLK1) labels a subset of dendritic microtubules and is required for KIF1-dependent dense-core vesicles ( DCVs) trafficking into dendrites and dendrite development. Our study demonstrates that microtubule-binding proteins can provide local signals for specific kinesin motors to drive polarized cargo transport. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the potential implication of microRNA on tuberous sclerosis (TSC) pathogenesis by performing microRNA profiling on cell lines silencing TSC1 or TSC2 genes using qPCR panels, before and after incubation with rapamycin. Significant differences in expression were observed between samples before and after rapamycin treatment in nineteen miRNAs in TSC1, five miRNAs in TSC2 and seven miRNAs in controls. Of miRNAs dysregulated before rapamycin treatment, three normalized after treatment in the TSC1 group (miR-21-3p, miR-433-3p, let-7g-3p) and one normalized in the TSC2 group (miR-1224-3p). Of the miRNAs dysregulated before rapamycin treatment in the TSC1 and TSC2 groups, two did not normalize after treatment (miR-33a-3p, miR-29a-3p). The results of the possible targets indicated that there are four common genes with seed regions susceptible to regulation by those miRNAs: ZBTB20, PHACTR2, PLXNC1 and ATP1B4. Our data show no changes in mRNA expression of these targets after rapamycin treatment. In conclusion, results of our study indicate the involvement of miRNA dysregulation in the pathogenesis of TSC. Some of the miRNA might be used as markers of treatment efficacy and autonomic miRNA as a target for future therapy. [ABSTRACT FROM AUTHOR]

Neurons are highly specialized for the processing and transmission of electrical signals and use cytoskeletonbased motor proteins to transport different vesicles and cellular materials. Abnormalities in intracellular transport are thought to be a critical factor in the degeneration and death of neurons in both the central and peripheral nervous systems. Several recent studies describe disruptive mutations in the minus-enddirected microtubule motor cytoplasmic dynein that are directly linked to human motor neuropathies, such as SMA (spinal muscular atrophy) and axonal CMT (Charcot-Marie-Tooth) disease or malformations of cortical development, including lissencephaly, pachygyria and polymicrogyria. In addition, genetic defects associated with these and other neurological disorders have been found in multifunctional adaptors that regulate dynein function, including the dynactin subunit p150Glued, BICD2 (Bicaudal D2), Lis-1 (lissencephaly 1) and NDE1 (nuclear distribution protein E). In the present paper we provide an overview of the diseasecausing mutations in dynein motors and regulatory proteins that lead to a broad phenotypic spectrum extending from peripheral neuropathies to cerebral malformations. [ABSTRACT FROM AUTHOR]

Alterations in social behavior are core symptoms of major developmental neuropsychiatric diseases such as autism spectrum disorders or schizophrenia. Hence, understanding their molecular and cellular underpinnings constitutes the major research task. Dysregulation of the global gene expression program in the developing brain leads to modifications in a number of neuronal connections, synaptic strength and shape, causing unbalanced neuronal plasticity, which may be important substrate in the pathogenesis of neurodevelopmental disorders, contributing to their clinical outcome. Serum response factor (SRF) is a major transcription factor in the brain. The behavioral influence of SRF deletion during neuronal differentiation and maturation has never been studied because previous attempts to knock-out the gene caused premature death. Herein, we generated mice that lacked SRF from early postnatal development to precisely investigate the role of SRF starting in the specific time window before maturation of excitatory synapses that are located on dendritic spine occurs. We show that the time-controlled loss of SRF in neurons alters specific aspects of social behaviors in SRF knock-out mice, and causes deficits in developmental spine maturation at both the structural and functional levels, including downregulated expression of the AMPARs subunits GluA1 and GluA2, and increases the percentage of filopodial/immature dendritic spines. In aggregate, our study uncovers the consequences of postnatal SRF elimination for spine maturation and social interactions revealing novel mechanisms underlying developmental neuropsychiatric diseases. [ABSTRACT FROM AUTHOR]

Background: Understanding of the molecular mechanisms of prefrontal cortex (PFC) plasticity is important for developing new treatment strategies for mental disorders such as depression and schizophrenia. Long-term potentiation (LTP) is a valid model for synaptic plasticity. The extracellular proteolytic system composed of matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) has recently been shown to play major role in the hippocampal plasticity. Methods: We tested whether induction of hippocampal-prefrontal LTP results in accumulation of tissue inhibitor of MMP-1, TIMP-1 mRNA, in the PFC of rats and whether adenovirally driven overexpression of TIMP-1 affects LTP. Additional study of slices was done with a specific MMP-9 inhibitor. Results: The TIMP-1 is induced in the rat medial PFC by stimuli evoking late LTP; its overexpression blocks the gelatinolytic activity of the MMP family; its overexpression before tetanization blocks late LTP in vivo; and MMP-9 inhibitor prevents late LTP in vitro. Conclusions: We suggest a novel extracellular mechanism of late LTP in the PFC, engaging TIMP-1-controlled proteolysis as an element of information integration. Our results may also be meaningful to an understanding of mental diseases and development of new treatment strategies that are based on extracellular mechanisms of synaptic plasticity. [Copyright &y& Elsevier]

Thiophene moieties were incorporated into previously described Zinspy (ZS) fluorescent Zn(II) sensor motifs (Nolan, E. M.; Lippard, S. J. Inorg. Chem. 2004, 43, 8310-8317) to provide enhanced fluorescence properties, low-micromolar dissociation constants for Zn(II), and improved Zn(II) selectivity. Halogenation of the xanthenone and benzoate moieties of the fluorescein platform systematically modulates the excitation and emission profiles, pH-dependent fluorescence, Zn(II) affinity, and Zn(II) complexation rates, offering a general strategy for tuning multiple properties of xanthenone-based metal ion sensors. Extensive biological studies in cultured cells and primary neuronal cultures demonstrate 2-{6-hydroxy-3- oxo-4,5-bis[(pyridin-2-ylmethylthiophen-2-ylmethylamino)methyl)-3H-xanthen-9-yl}benzoic acid (ZS5) to be a versatile imaging tool for detecting Zn(II) in vivo. ZS5 localizes to the mitochondria of HeLa cells and allows visualization of glutamate-mediated Zn(II) uptake in dendrites and Zn(II) release resulting from nitrosative stress in neurons. [ABSTRACT FROM AUTHOR]

Estrogens play an important role in the brain function acting through two receptor types, ERα and ERβ, both well-recognized as transcription factors. In this study, we investigated the ERβ mRNA and protein levels in the rat hippocampus by using two in vivo models that are known to affect synapse formation. Natural estrous-proestrous cycle was used as a model in which a marked decrease in the density of hippocampal synapses was previously observed between proestrus and estrus. We have found that ERβ mRNA and protein were displayed in high levels in the estrus and in low levels in the proestrous phase. By applying kainic acid (KA) to adult rats, we demonstrated that up-regulation of ERβ mRNA and protein in hippocampal CA regions was vulnerable to KA-induced excitotoxicity. Furthermore, we note a concomitant decrease of ERβ in the excitotoxicity-resistant denate gyrus that undergoes intense plastic changes, including synaptogenesis. These data suggested that decreases in ERβ expression correlated with increase in synapse formation. This notion has been tested in vitro in hippocampal cultures, in which overexpression of ERβ by means of gene transfection resulted in the lowering of the dendritic spine density that was elevated by estrogen. In summary, our results suggest that ERβ inhibits synapse formation in hippocampal neurons. © 2006 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

The synthesis of a difluorofluorescein monocarboxaldehyde platform and its use for preparing ZP8, a new member of the Zinpyr family of neuronal Zn2+ sensors, are described. By combining an aniline photoinduced electron transfer (PET) switch and an electron-withdrawing fluorescein scaffold, ZP8 displays reduced background fluorescence and improved dynamic range compared to previous ZP probes. The bright sensor undergoes an 11-fold increase in fluorescence intensity upon Zn2+ complexation (Φ = 0.03-0.35) with high selectivity over cellular concentrations of Ca2+ and Mg2+. In addition, sensors in the ZP family have been utilized for optical imaging in biological samples using two-photon microscopy (TPM). The cell-permeable ZP3 probe is capable of identifying natural pools of labile Zn2+ within the mossy fiber synapses of live hippocampal slices using TPM, establishing the application of this technique for monitoring endogenous Zn2+ stores. [ABSTRACT FROM AUTHOR]

A series of new fluorescent Zinpyr (ZP) chemosensors based on the fluorescein platform have been prepared and evaluated for imaging neuronal Zn2+. A systematic synthetic survey of electronegative substitution patterns on a homologous ZP scaffold provides a basis for tuning the fluorescence responses of “off-on” photoinduced electron transfer (PET) probes by controlling fluorophore pKa values and attendant proton-induced interfering fluorescence of the metal-free (apo) probes at physiological pH. We further establish the value of these improved optical tools for interrogating the metalloneurochemistry of Zn2+; the novel ZP3 fluorophore images endogenous stores of Zn2+ in live hippocampal neurons and slices, including the first fluorescence detection of Zn2+ in isolated dentate gyrus cultures. Our findings reveal that careful control of fluorophore pKa can minimize proton-induced fluorescence of the apo probes and that electronegative substitution offers a general strategy for tuning PET chemosensors for cellular studies. In addition to providing improved optical tools for Zn2+ in the neurosciences, these results afford a rational starting point for creating superior fluorescent probes for biological applications. [Copyright &y& Elsevier]

Neurons are exposed to damaging stimuli that can trigger cell death and subsequently cause serious neurological disorders. Therefore, it is important to define defense mechanisms that can be activated in response to damage to reduce neuronal loss. Here we report that cisplatin (CPDD), a neurotoxic anticancer drug that damages DNA, triggered apoptosis and activated the extracellular signal-regulated kinase ½ (ERK½) pathway in cultured rat cortical neurons. Inhibition of ERK½ activation using either pharmacological inhibitors or a dominant-negative mutant of the ERK½ activator, mitogen-activated protein kinase kinase 1, increased the toxicity of CPDD. Interestingly, N-methyl-Daspartate (NMDA) receptor (NMDAR) antagonists reduced the ERK½ activation and exacerbated apoptosis in CPDD-treated neurons. Pre-treatment with CPDD increased ERK½ activation triggered by exogenous NMDA, suggesting that CPDD augmented NMDAR responsiveness. CPDD-enhanced response of NMDAR and CPDD-mediated ERK½ activation were both decreased by inhibition of poly(ADP-ribose) polymerase (PARP). Interestingly, PARP activation did not produce ATP depletion, suggesting involvement of a non-energetic mechanism in NMDAR regulation by PARP. Finally, CPDD toxicity was reduced by brain-derived neurotrophic factor, and this protection required ERK½. In summary, our data identify a novel compensatory circuit in central nervous system neurons that couples the DNA injury, through PARP and NMDAR, to the defensive ERK½ activation. [ABSTRACT FROM AUTHOR]

One of the characteristic features of different classes of neurons that is vital for their proper functioning within neuronal networks is the shape of their dendritic arbors. To properly develop dendritic trees, neurons need to accurately control the intracellular transport of various cellular cargo (e.g., mRNA, proteins, and organelles). Microtubules and motor proteins (e.g., dynein and kinesins) that move along microtubule tracks play an essential role in cargo sorting and transport to the most distal ends of neurons. Equally important are motor adaptors, which may affect motor activity and specify cargo that is transported by the motor. Such transport undergoes very dynamic fine‐tuning in response to changes in the extracellular environment and synaptic transmission. Such regulation is achieved by the phosphorylation of motors, motor adaptors, and cargo, among other mechanisms. This review focuses on the contribution of the dynein–dynactin complex, kinesins, their adaptors, and the phosphorylation of these proteins in the formation of dendritic trees by maturing neurons. We primarily review the effects of the motor activity of these proteins in dendrites on dendritogenesis. We also discuss less anticipated mechanisms that contribute to dendrite growth, such as dynein‐driven axonal transport and non‐motor functions of kinesins. [ABSTRACT FROM AUTHOR]

Tuberous Sclerosis Complex (TSC) is a rare genetic disease that manifests with early symptoms, including cortical malformations, childhood epilepsy, and TSC-associated neuropsychiatric disorders (TANDs). Cortical malformations arise during embryonic development and have been linked to childhood epilepsy before, but the underlying mechanisms of this relationship remain insufficiently understood. Zebrafish have emerged as a convenient model to study elementary neurodevelopment; however, without in-depth functional analysis, the Tsc2-deficient zebrafish line cannot be used for studies of TANDs or new drug screening. In this study, we found that the lack of Tsc2 in zebrafish resulted in heterotopias and hyperactivation of the mTorC1 pathway in pallial regions, which are homologous to the mammalian cortex. We observed commissural thinning that was responsible for brain dysconnectivity, recapitulating TSC pathology in human patients. The lack of Tsc2 also delayed axonal development and caused aberrant tract fasciculation, corresponding to the abnormal expression of genes involved in axon navigation. The mutants underwent epileptogenesis that resulted in nonmotor seizures and exhibited increased anxiety-like behavior. We further mapped discrete parameters of locomotor activity to epilepsy-like and anxiety-like behaviors, which were rescued by reducing tyrosine receptor kinase B (TrkB) signaling. Moreover, in contrast to treatment with vigabatrin and rapamycin, TrkB inhibition rescued brain dysconnectivity and anxiety-like behavior. These data reveal that commissural thinning results in the aberrant regulation of anxiety, providing a mechanistic link between brain anatomy and human TANDs. Our findings also implicate TrkB signaling in the complex pathology of TSC and reveal a therapeutic target. [ABSTRACT FROM AUTHOR]

The angiomotin (Amot)–Yes-associated protein 1 (Yap1) complex plays a major role in regulating the inhibition of cell contact, cellular polarity, and cell growth in many cell types. However, the function of Amot and the Hippo pathway transcription coactivator Yap1 in the central nervous system remains unclear. We found that Amot is a critical mediator of dendritic morphogenesis in cultured hippocampal cells and Purkinje cells in the brain. Amot function in developing neurons depends on interactions with Yap1, which is also indispensable for dendrite growth and arborization in vitro. The conditional deletion of Amot and Yap1 in neurons led to a decrease in the complexity of Purkinje cell dendritic trees, abnormal cerebellar morphology, and impairments in motor coordination. Our results indicate that the function of Amot and Yap1 in dendrite growth does not rely on interactions with TEA domain (TEAD) transcription factors or the expression of Hippo pathway–dependent genes. Instead, Amot and Yap1 regulate dendrite development by affecting the phosphorylation of S6 kinase and its target S6 ribosomal protein. [ABSTRACT FROM AUTHOR]

Mammalian/mechanistic target of rapamycin (mTOR) is a serine-threonine kinase that controls several important aspects of mammalian cell function. mTOR activity is modulated by various intra- and extracellular factors; in turn, mTOR changes rates of translation, transcription, protein degradation, cell signaling, metabolism, and cytoskeleton dynamics. mTOR has been repeatedly shown to participate in neuronal development and the proper functioning of mature neurons. Changes in mTOR activity are often observed in nervous system diseases, including genetic diseases (e.g., tuberous sclerosis complex, Pten-related syndromes, neurofibromatosis, and Fragile X syndrome), epilepsy, brain tumors, and neurodegenerative disorders (Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease). Neuroscientists only recently began deciphering the molecular processes that are downstream of mTOR that participate in proper function of the nervous system. As a result, we are gaining knowledge about the ways in which aberrant changes in mTOR activity lead to various nervous system diseases. In this review, we provide a comprehensive view of mTOR in the nervous system, with a special focus on the neuronal functions of mTOR (e.g., control of translation, transcription, and autophagy) that likely underlie the contribution of mTOR to nervous system diseases. [ABSTRACT FROM AUTHOR]

Tuberous sclerosis complex (TSC) is a rare multi-system disorder, primary manifestations of which are benign tumors and lesions in various organs of the body, including the brain. TSC patients often suffer from epilepsy, mental retardation, and autism spectrum disorder (ASD). Therefore, TSC serves as a model of epilepsy, ASD, and tumorigenesis. TSC is caused by the lack of functional Tsc1-Tsc2 complex, which serves as a major cellular inhibitor of mammalian Target of Rapamycin Complex 1 (mTORC1). mTORC1 is a kinase controlling most of anabolic processes in eukaryotic cells. Consequently, mTORC1 inhibitors, such as rapamycin, serve as experimental or already approved drugs for several TSC symptoms. However, rapalogs, although quite effective, need to be administered chronically and likely for a lifetime, since therapy discontinuation results in tumor regrowth and epilepsy recurrence. Recent studies revealed that metabolism and excitability (in the case of neurons) of cells lacking Tsc1-Tsc2 complex are changed, and these features may potentially be used to treat some of TSC symptoms. In this review, we first provide basic facts about TSC and its molecular background, to next discuss the newest findings in TSC cell biology that can be used to improve existing therapies of TSC and other diseases linked to mTORC1 hyperactivation. © 2016 IUBMB Life, 68(12):955-962, 2016 [ABSTRACT FROM AUTHOR]

The article focuses on the significance of oganelle-specific zinc detection using zinpyr-labeled fusion proteins in live cells. It notes that the applications of zinc-selective fluorescent sensors will help construct a detailed network map of biological zinc and its transporter proteins and to clarify how zinc influences processes including neurotransmission and proliferative signaling. Opportunities offered by labeling techniques could provide valuable tools to illuminate zinc trafficking.