Factor Xa subsite mapping by proteome-derived peptide libraries improved using WebPICS, a resource for proteomic identification of cleavage sites

Proteomic identification of protease cleavage site specificity (PICS) is a recent proteomic approach for the easy mapping of protease subsite preferences that determines both the prime- and non-prime side specificity concurrently. Here we greatly facilitate user access by providing an automated and simple web-based data-analysis resource termed WebPics (http://clipserve.clip.ubc.ca/pics/). We demonstrate the utility of WebPics analysis of PICS data by determining the substrate specificity of factor Xa from P6-P6', an important blood coagulation protease that proteolytically generates thrombin from prothrombin. PICS confirms existing data on non-prime site specificity and refines our knowledge of factor Xa prime-site selectivit

CD14 Counterregulates Lipopolysacharide- Induced Tumor Necrosis Factor-α Production in a Macrophage Subset

In response to GM-CSF or M-CSF, macrophages (MΦ) can acquire pro- or anti-inflammatory properties, respectively. Given the importance of CD14 and Toll-like receptor (TLR) 4 in lipopolysaccharide (LPS)-induced signaling, we studied the effect of anti-CD14 antibody mediated CD14 blockade on LPS-induced cytokine production, signal transduction and on the expression levels of CD14 and TLR4 in GM-MΦ and M-MΦ. We found M-MΦ to express higher levels of both surface antigens and to produce more interferon (IFN)-β and interleukin-10, but less tumor necrosis factor (TNF)-α than GM-MΦ. Blockage of CD14 at high LPS concentrations increased the production of proinflammatory cytokines and decreased that of IFN-β in M-MΦ but not in GM-MΦ. We show that phosphorylation states of signaling molecules of the MyD88 (myeloid differentiation primary response 88), TRIF (TIR-domain-containing adapter-inducing IFN-β) and MAPK (mitogen-activated protein kinase) pathways are not altered in any way that would account for the cytokine overshoot reaction. However, CD14 blockage in M-MΦ decreased TLR4 and CD14 expression levels, regardless of the presence of LPS, indicating that the loss of the surface molecules prevented LPS from initiating TRIF signaling. As TNF-α synthesis was even upregulated under these experimental conditions, we suggest that TRIF is normally involved in restricting LPSinduced TNF-α overproduction. Thus, surface CD14 plays a decisive role in the biological response by determining LPSinduced signaling

Evidence for a large off-centered galactic outflow and its connection to the extraplanar diffuse ionized gas in IC 1553

Aims. We analyze a MUSE optical integral field spectrum of the star-forming edge-on galaxy IC 1553 in order to study its extraplanar diffuse ionized gas (eDIG) and the processes shaping its disk-halo interface. Methods. We extracted the optical emission line properties from the integral field spectrum and generated the commonly used emission line diagnostic diagrams in order to analyze the ionization conditions and the distribution of the eDIG. Furthermore, we performed gravitational potential fitting to investigate the kinematics of a suspected galactic outflow. Results. We find that the eDIG scale height has a maximum value of approximately 1.0 kpc and decreases roughly linearly with the radial distance from the galactic center in projection. The ionization state of the eDIG is not consistent with a pure photoionization scenario and instead requires a significant contribution from shock ionization. This, in addition to the gas kinematics, strongly suggests the presence of a galactic scale outflow, the origin of which lies at least 1.4 kpc away from the galactic center. The inferred shock velocity in the eDIG of approximately 225 km s-1 is comparable to the escape velocity estimated from our potential modelling. The asymmetric distribution of currently star-forming clusters produces a range of different ionization conditions in the eDIG. As a result, the vertical emission line profiles vary quantitatively and qualitatively along the major axis of the galaxy. This analysis illustrates that it is crucial in studies of the eDIG to use observations that take the spatial and kinematical distributions into account, such as those done with integral field units, to form an accurate picture of the relevant physical properties.Comment: Accepted for publication by Astronomy & Astrophysics on August 1, 202

Passive Immunization against Pyroglutamate-3 Amyloid-β Reduces Plaque Burden in Alzheimer-Like Transgenic Mice: A Pilot Study

Background: N-terminally truncated and modified pyroglutamate-3 amyloid-β protein (pE3-Aβ) is present in most, if not all, cerebral plaque and vascular amyloid deposits in human Alzheimer's disease (AD). pE3-Aβ deposition is also found in AD-like transgenic (tg) mouse brain, albeit in lesser quantities than general Aβ. pE3-Aβ resists degradation, is neurotoxic, and may act as a seed for Aβ aggregation. Objective: We sought to determine if pE3-Aβ removal by passive immunization with a highly specific monoclonal antibody (mAb) impacts pathogenesis in a mouse model of Alzheimer's amyloidosis. Methods: APPswe/PS1ΔE9 tg mice were given weekly intraperitoneal injections of a new anti-pE3-Aβ mAb (mAb07/1) or PBS from 5.8 to 13.8 months of age (prevention) or from 23 to 24.7 months of age (therapeutic). Multiple forms of cerebral Aβ were quantified pathologically and biochemically. Gliosis and microhemorrhage were examined. Results: Chronic passive immunization with an anti-pE3-Aβ mAb significantly reduced total plaque deposition and appeared to lower gliosis in the hippocampus and cerebellum in both the prevention and therapeutic studies. Insoluble Aβ levels in hemibrain homogenates were not significantly different between immunized and control mice. Microhemorrhage was not observed with anti-pE3-Aβ immunotherapy. Conclusions: Selective removal of pE3-Aβ lowered general Aβ plaque deposition suggesting a pro-aggregation or seeding role for pE3-Aβ

The Advocate

Headlines Include: Laurels For Feerick: An Alumnus To Remember; Crime at Fordham; Who\u27s Next?, Film at 11https://ir.lawnet.fordham.edu/student_the_advocate/1007/thumbnail.jp

Depression Diagnosis using Deep Convolutional Neural Networks

Depression is a prevalent psychiatric disorder that impacts the quality of life of 300 million people around the world. The complex nature of depression manifestations in patients and the lack of technological advances in the diagnosis process has left a lot of room for improvement in this particular domain. At present, the diagnosis is mainly made by physicians during a conversation comprising the exploration of the symptoms and the diagnostic criteria for depression. Recently, the electroencephalography (EEG) has regained interest as a promising approach to provide bio-markers which are of clinical value in the diagnostic process and for response prediction to therapy. In the present landscape, even the addition of EEG data has resulted in a semi-automated process, where the expert still has to heavily modify the raw data. This adds an inherent bias to the process based on the expert and incurs costs as well as time to the process of diagnosis. In this paper, we present a fast, effective and automated method that is able to quickly determine if the patient has depression while still maintaining a high accuracy of diagnosis. Our approach is built on using raw EEG-data, performing frequency domain preprocessing in order to split the data into its different frequency domains and to create EEG ’images’. These images are then treated by a convolutional neural network, which is a novel approach in this area. Experimental results have shown to provide outstanding results and to work without the need for feature engineering or any human interaction, which is a core strength of the model we are proposing

Glutaminyl cyclase contributes to the formation of focal and diffuse pyroglutamate (pGlu)-Aβ deposits in hippocampus via distinct cellular mechanisms

In the hippocampal formation of Alzheimer’s disease (AD) patients, both focal and diffuse deposits of Aβ peptides appear in a subregion- and layer-specific manner. Recently, pyroglutamate (pGlu or pE)-modified Aβ peptides were identified as a highly pathogenic and seeding Aβ peptide species. Since the pE modification is catalyzed by glutaminyl cyclase (QC) this enzyme emerged as a novel pharmacological target for AD therapy. Here, we reveal the role of QC in the formation of different types of hippocampal pE-Aβ aggregates. First, we demonstrate that both, focal and diffuse pE-Aβ deposits are present in defined layers of the AD hippocampus. While the focal type of pE-Aβ aggregates was found to be associated with the somata of QC-expressing interneurons, the diffuse type was not. To address this discrepancy, the hippocampus of amyloid precursor protein transgenic mice was analysed. Similar to observations made in AD, focal (i.e. core-containing) pE-Aβ deposits originating from QC-positive neurons and diffuse pE-Aβ deposits not associated with QC were detected in Tg2576 mouse hippocampus. The hippocampal layers harbouring diffuse pE-Aβ deposits receive multiple afferents from QC-rich neuronal populations of the entorhinal cortex and locus coeruleus. This might point towards a mechanism in which pE-Aβ and/or QC are being released from projection neurons at hippocampal synapses. Indeed, there are a number of reports demonstrating the reduction of diffuse, but not of focal, Aβ deposits in hippocampus after deafferentation experiments. Moreover, we demonstrate in neurons by live cell imaging and by enzymatic activity assays that QC is secreted in a constitutive and regulated manner. Thus, it is concluded that hippocampal pE-Aβ plaques may develop through at least two different mechanisms: intracellularly at sites of somatic QC activity as well as extracellularly through seeding at terminal fields of QC expressing projection neurons

I-BEAT: New ultrasonic method for single bunch measurement of ion energy distribution

The shape of a wave carries all information about the spatial and temporal structure of its source, given that the medium and its properties are known. Most modern imaging methods seek to utilize this nature of waves originating from Huygens' principle. We discuss the retrieval of the complete kinetic energy distribution from the acoustic trace that is recorded when a short ion bunch deposits its energy in water. This novel method, which we refer to as Ion-Bunch Energy Acoustic Tracing (I-BEAT), is a generalization of the ionoacoustic approach. Featuring compactness, simple operation, indestructibility and high dynamic ranges in energy and intensity, I-BEAT is a promising approach to meet the needs of petawatt-class laser-based ion accelerators. With its capability of completely monitoring a single, focused proton bunch with prompt readout it, is expected to have particular impact for experiments and applications using ultrashort ion bunches in high flux regimes. We demonstrate its functionality using it with two laser-driven ion sources for quantitative determination of the kinetic energy distribution of single, focused proton bunches.Comment: Paper: 17 Pages, 3 figures Supplementary Material 16 pages, 7 figure

Klinische Performance eines neuen SARS-CoV-2-Antigen-Tests in der Notaufnahme eines Maximalversorgers

Ein Baustein zur Eindämmung der COVID-19-Pandemie ist die Verfügbarkeit von Tests mit hoher Sensitivität und Spezifität zur Detektion von SARS-CoV-2, insbesondere um Infizierte in vulnerablen Einrichtungen, z. B. Krankenhäusern und Pflegeeinrichtungen, zeitnah identifizieren und isolieren zu können. Dies betrifft alle Personengruppen dieser Einrichtungen, also Patient*innen/Bewohner*innen, Besucher*innen als auch Personal. Bisheriger Goldstandard für den Nachweis einer SARS-CoV-2-Infektion ist die RT-PCR. SARS-CoV-2-Antigen-Tests sind aufgrund ihres Point-of-Care-Ansatzes, der einfachen Handhabung und des günstigeren Preises eine wertvolle Ergänzung zur RT-PCR-Diagnostik. Sie erkennen mit ausreichender Sicherheit SARS-CoV-2-Infektionen bei symptomatischen Patient*innen und in Proben mit niedrigen Ct-Werten in der RT-PCR. Als Einzeltestung bei asymptomatischen Patient*innen ist ihre Wertigkeit dagegen deutlich eingeschränkt. Hier sollten repetitive Antigen-Testungen oder primär PCR-basierte Verfahren zur Anwendung kommen