Neuroacanthocytosis Syndromes: The Clinical Perspective

The two very rare neurodegenerative diseases historically known as the “neuroacanthocytosis syndromes” are due to mutations of either VPS13A or XK. These are phenotypically similar disorders that affect primarily the basal ganglia and hence result in involuntary abnormal movements as well as neuropsychiatric and cognitive alterations. There are other shared features such as abnormalities of red cell membranes which result in acanthocytes, whose relationship to neurodegeneration is not yet known. Recent insights into the functions of these two proteins suggest dysfunction of lipid processing and trafficking at the subcellular level and may provide a mechanism for neuronal dysfunction and death, and potentially a target for therapeutic interventions

P3-[2-(4-hydroxyphenyl)-2-oxo]ethyl ATP for the Rapid Activation of the Na+,K+-ATPase

This is the published version, also available here: http://dx.doi.org/10.1016/S0006-3495(00)76387-9.P3-[2-(4-hydroxyphenyl)-2-oxo]ethyl ATP (pHP-caged ATP) has been investigated for its application as a phototrigger for the rapid activation of electrogenic ion pumps. The yield of ATP after irradiation with a XeCl excimer laser (λ = 308 nm) was determined at pH 6.0–7.5. For comparison, the photolytic yields of P3-[1-(2-nitrophenyl)]ethyl ATP (NPE-caged ATP) and P3-[1,2-diphenyl-2-oxo]ethyl ATP (desyl-caged ATP) were also measured. It was shown that at λ = 308 nm pHP-caged ATP is superior to the other caged ATP derivatives investigated in terms of yield of ATP after irradiation. Using time-resolved single-wavelength IR spectroscopy, we determined a lower limit of 106 s−1 for the rate constant of release of ATP from pHP-caged ATP at pH 7.0. Like NPE-caged ATP, pHP-caged ATP and desyl-caged ATP bind to the Na+,K+-ATPase and act as competitive inhibitors of ATPase function. Using pHP-caged ATP, we investigated the charge translocation kinetics of the Na+,K+-ATPase at pH 6.2–7.4. The kinetic parameters obtained from the electrical measurements are compared to those obtained with a technique that does not require caged ATP, namely parallel stopped-flow experiments using the voltage-sensitive dye RH421. It is shown that the two techniques yield identical results, provided the inhibitory properties of the caged compound are taken into account. Our results demonstrate that under physiological (pH 7.0) and slightly basic (pH 7.5) or acidic (pH 6.0) conditions, pHP-caged ATP is a rapid, effective, and biocompatible phototrigger for ATP-driven biological systems

Flux Dendrites of Opposite Polarity in Superconducting MgB2_2 rings observed with magneto-optical imaging

Magneto-optical imaging was used to observe flux dendrites with opposite polarities simultaneously penetrate superconducting, ring-shaped MgB$_2$ films. By applying a perpendicular magnetic field, branching dendritic structures nucleate at the outer edge and abruptly propagate deep into the rings. When these structures reach close to the inner edge, where flux with opposite polarity has penetrated the superconductor, they occasionally trigger anti-flux dendrites. These anti-dendrites do not branch, but instead trace the triggering dendrite in the backward direction. Two trigger mechanisms, a non-local magnetic and a local thermal, are considered as possible explanations for this unexpected behaviour. Increasing the applied field further, the rings are perforated by dendrites which carry flux to the center hole. Repeated perforations lead to a reversed field profile and new features of dendrite activity when the applied field is subsequently reduced.Comment: 6 pages, 6 figures, accepted to Phys. Rev.

Structure-Based Screening of Tetrazolylhydrazide Inhibitors versus KDM4 Histone Demethylases

Human histone demethylases are known to play an important role in the development of several tumor types. Consequently, they have emerged as important medical targets for the treatment of human cancer. Herein, structural studies on tetrazolylhydrazide inhibitors as a new scaffold for a certain class of histone demethylases, the JmjC proteins, are reported. A series of compounds are structurally described and their respective binding modes to the KDM4D protein, which serves as a high-resolution model to represent the KDM4 subfamily in crystallographic studies, are examined. Similar to previously reported inhibitors, the compounds described herein are competitors for the natural KDM4 cofactor, 2-oxoglutarate. The tetrazolylhydrazide scaffold fills an important gap in KDM4 inhibition and newly described, detailed interactions of inhibitor moieties pave the way to the development of compounds with high target-binding affinity and increased membrane permeability, at the same time

A petri nets based design of cognitive radios using distributed signal processing

AbstractReconfigurability for transceivers for wireless access networks like Bluetooth, WiMAX and W-LAN will become increasingly important. An appropriately flexible and reliable software architecture, allowing the concurrent processing of different controlling tasks for wireless terminals will hence be an important asset. Already during the 1980s reconfigurable receivers were developed for radio intelligence in the short wave range and the concept of software radio (SR) was born. A software defined radio (SDR) is a practical version of an SR: The received signals are sampled after a suitable band selection filter, usually in the base band or a low intermediate frequency band. The signal processing in both SR and SDR requires a considerable amount of concurrent processes. Since Petri nets (PNs) are both simple and strong tools for the description and the design of such concurrent processes, it is recommendable to deploy them for SDR. SDRs have paved the way towards cognitive radios (CRs), which are based on SDRs that additionally sense their environments, track changes, and react upon their findings. A CR is an autonomous unit in a communications environment that frequently exchanges information with the networks it is able to access as well as with other CRs. In this communication, the authors will introduce a realization concept for a CR which forms the basis of a hardware/firmware demonstrator developed by the authors. This demonstrator makes use of a digital signal processor (DSP) which forms the core of the design and flexibly programmable hardware accelerators based on field programmable gate arrays (FPGAs). The authors will describe the solution also in view of the recent developments of IEEE 802.2

Characterization of Fluorescent Proteins with Intramolecular Photostabilization*

Genetically encodable fluorescent proteins have revolutionized biological imaging in vivo and in vitro. Despite their importance, their photophysical properties, i. e., brightness, count-rate and photostability, are relatively poor compared to synthetic organic fluorophores or quantum dots. Intramolecular photostabilizers were recently rediscovered as an effective approach to improve photophysical properties of organic fluorophores. Here, direct conjugation of triplet-state quenchers or redox-active substances creates high local concentrations of photostabilizer around the fluorophore. In this paper, we screen for effects of covalently linked photostabilizers on fluorescent proteins. We produced a double cysteine mutant (A206C/L221C) of α-GFP for attachment of photostabilizer-maleimides on the β-barrel near the chromophore. Whereas labelling with photostabilizers such as trolox, a nitrophenyl group, and cyclooctatetraene, which are often used for organic fluorophores, had no effect on α-GFP-photostability, a substantial increase of photostability was found upon conjugation to azobenzene. Although the mechanism of the photostabilizing effects remains to be elucidated, we speculate that the higher triplet-energy of azobenzene might be crucial for triplet-quenching of fluorophores in the blue spectral range. Our study paves the way for the development of fluorescent proteins with photostabilizers in the protein barrel by methods such as unnatural amino acid incorporation. © 2021 The Authors. ChemBioChem published by Wiley-VCH Gmb

Characterization of fluorescent proteins with intramolecular photostabilization

Genetically encodable fluorescent proteins have revolutionized biological imaging in vivo and in vitro. Since there are no other natural fluorescent tags with comparable features, the impact of fluorescent proteins for biological research cannot be overemphasized. Despite their importance, their photophysical properties, i.e., brightness, count-rate and photostability, are relatively poor compared to synthetic organic fluorophores or quantum dots. Intramolecular photostabilizers were recently rediscovered as an effective approach to improve photophysical properties. The approach uses direct conjugation of photostablizing compounds such as triplet-state quenchers or redox-active substances to an organic fluorophore, thereby creating high local concentrations of photostabilizer. Here, we introduce an experimental strategy to screen for the effects of covalently-linked photostabilizers on fluorescent proteins. We recombinantly produced a double cysteine mutant (A206C/L221C) of α-GFP for attachment of photostabilizer-maleimides on the ß-barrel in close proximity to the chromophore. Whereas labelling with photostabilizers such as Trolox, Nitrophenyl, and Cyclooctatetraene, which are often used for organic fluorophores, had no effect on α-GFP-photostability, a substantial increase of photostability was found upon conjugation of α-GFP to an azobenzene derivative. Although the mechanism of the photostabilizing effects remains to be elucidated, we speculate that the higher triplet-energy of azobenzene might be crucial for triplet-quenching of fluorophores in the near-UV and blue spectral range. Our study paves the way towards the development and design of a second generation of fluorescent proteins with photostabilizers placed directly in the protein barrel by methods such as unnatural amino acid incorporation

Systematic Heterogeneity of Fractional Vesicle Pool Sizes and Release Rates of Hippocampal Synapses

AbstractHippocampal neurons in tissue culture develop functional synapses that exhibit considerable variation in synaptic vesicle content (20–350 vesicles). We examined absolute and fractional parameters of synaptic vesicle exocytosis of individual synapses. Their correlation to vesicle content was determined by activity-dependent discharge of FM-styryl dyes. At high frequency stimulation (30 Hz), synapses with large recycling pools released higher amounts of dye, but showed a lower fractional release compared to synapses that contained fewer vesicles. This effect gradually vanished at lower frequencies when stimulation was triggered at 20 Hz and 10 Hz, respectively. Live-cell antibody staining with anti-synaptotagmin-1-cypHer 5, and overexpression of synaptopHluorin as well as photoconversion of FM 1-43 followed by electron microscopy, consolidated the findings obtained with FM-styryl dyes. We found that the readily releasable pool grew with a power function with a coefficient of 2/3, possibly indicating a synaptic volume/surface dependency. This observation could be explained by assigning the rate-limiting factor for vesicle exocytosis at high frequency stimulation to the available active zone surface that is proportionally smaller in synapses with larger volumes

Impact of the gate geometry on adiabatic charge pumping in InAs double quantum dots

We compare the adiabatic quantized charge pumping performed in two types of InAs nanowire double quantum dots (DQDs), either with tunnel barriers defined by closely spaced narrow bottom gates, or by well-separated side gates. In the device with an array of bottom gates of 100 nm pitch and 10 mu m lengths, the pump current is quantized only up to frequencies of a few MHz due to the strong capacitive coupling between the bottom gates. In contrast, in devices with well-separated side gates with reduced mutual gate capacitances, we find well-defined pump currents up to 30 MHz. Our experiments demonstrate that high frequency quantized charge pumping requires careful optimization of the device geometry, including the typically neglected gate feed lines

Association of Hepatitis C Virus—Specific CD8+ T Cells with Viral Clearance in Acute Hepatitis C

CD8+ T lymphocytes play a major role in antiviral immune defense. Their significance for acute hepatitis C is unclear. Our aim was to correlate the CD8+ T cell response with the outcome of infection. Eighteen patients with acute hepatitis C and 19 normal donors were studied. Hepatitis C virus (HCV)—specific CD8+ T cells were identified in the enzyme-linked immunospot assay by their interferon-γ (IFN-γ) production after specific stimulation. The highest numbers of IFN-γ—producing HCV-specific CD8+ T cells were found in patients with acute hepatitis C and a self-limited course of disease during the first 6 months after onset of disease, but these numbers dropped thereafter to undetectable levels. The differences in responsiveness between patients with self-limited disease versus patients with a chronic course were statistically significant (P < .001). Our data show that the number of IFN—γ-producing HCV-specific CD8+ T cells during the first 6 months after onset of disease is associated with eradication of the HCV infectio