Einfluss des Glykoprotein B aus Herpes Simplex Virus Typ 1 auf die intrazelluläre Verteilung und Exozytose von HLA Klasse II-Molekülen

Herpes-Simplex Virus Typ-1 (HSV-1) ist ein weltweit verbreitetes humanpathogenes Virus, welches nach der Primärinfektion lebenslang im Wirtsorganismus verbleibt. Die Persistenz von HSV-1 im Organismus wird durch Manipulation der Immunantwort erreicht. Das HSV-1 kodierte Hüllprotein Glykoprotein B (gB) ist an der Bindung und der Fusion des Virus mit der Zielzelle beteiligt. In der vorliegenden Arbeit wurde der Einfluss von gB auf den intrazellulären Transport und auf die intrazelluläre Verteilung von HLA-DR-Molekülen (DR) untersucht. Da HSV-1 ein lytisches Virus ist und infizierte Zellen nur eine kurze Lebensspanne haben, wurde eine humane Melanomzellline (MJ), welche DR und die invariante Kette (Ii) exprimiert, stabil mit gB transfiziert (MJ-gB). Mit Hilfe dieser Zellen konnte gezeigt werden, dass gB mit DR interagiert und dass Ii und gB um die Bindung an DR-Moleküle kompetieren. Da innerhalb des endoplasmatischen Retikulums (ER) ein Überschuss an Ii vorliegt, erfolgt die Assoziation von gB an DR nicht im ER, sondern erst nach Degradation von Ii in den MHC Klasse II-Beladungskompartimenten (MIICs). Im Gegensatz zu freien DR-Molekülen werden gB-assoziierte DR-Moleküle innerhalb der MIICs nicht mit hochaffinen Peptiden beladen. In der vorliegenden Arbeit wurde gezeigt, dass MJ-gB-Zellen vergrößerte endosomale Kompartimente besitzen. Diese Kompartimente enthalten gB, DR-Moleküle und das Tetraspannin CD63, einen Marker für MIICs. Nach der Fusion der limitierenden Membran der MIICs mit der Plasmamembran werden DR-positive intraluminale Vesikel in den extrazellulären Raum freigesetzt. Aus diesen Vesikeln, die auch als Exosomen bezeichnet werden, konnten gB-MHCII-Komplexe isoliert werden. Es konnte zudem gezeigt werden, dass MJ-gB-Zellen insgesamt eine größere Menge an DR-Molekülen und CD63 über Exosomen freisetzen als MJ-Zellen. Ein Mechanismus für die Sortierung von Proteinen in MVEs bzw. Exosomen ist eine Modifikation mit Ubiquitin. Ubiquitinylierte Proteine konnten in den vergrößerten Kompartimenten der MJ-gB-Zellen nachgewiesen werden. Es wurde auch gezeigt, dass aus MJ-gB-Zellen isoliertes gB, nicht aber DR-Untereinheiten ubiquitinyliert sind. Aus diesen Daten kann folgendes Modell abgeleitet werden: In MJ-gB-Zellen verursacht gB eine Vergrößerung endosomaler Vesikel, in denen sich gB, DR, gB-DR-Komplexe, CD63 und ubiquitinylierte Proteine anhäufen. In diesen Kompartimenten werden gB-DR-Komplexe und CD63 in die intraluminalen Vesikel sortiert, verbleiben aber nicht in der limitierenden Membran der MIICs, welche später mit der Plasmamembran fusioniert. Nach Fusion der MIICs mit der Zellmembran, werden gB-DR-Komplexe enthaltende Exosomen in den extrazellulären Raum freigesetzt. In dieser Arbeit wurde ein bisher unbekannter Weg der Immunevasion von HSV-1 entdeckt. Bei diesem gB-vermittelten Mechanismus werden DR-Moleküle durch gB gezielt in Exosomen sortiert und in das extrazelluläre Milieu freigesetzt

Invasive Mold Infection of the Central Nervous System in Immunocompromised Children

Background: Due to the difficulties in the definite diagnosis, data on brain imaging in pediatric patients with central nervous system (CNS)-invasive mold infection (IMD) are scarce. Our aim was to describe brain imaging abnormalities seen in immunocompromised children with CNS-IMD, and to analyze retrospectively whether specific imaging findings and sequences have a prognostic value. Methods: In a retrospective study of 19 pediatric patients with proven or probable CNS-IMD, magnetic resonance imaging (MRI)-findings were described and analyzed. The results were correlated with outcome, namely death, severe sequelae, or no neurological sequelae. Results: 11 children and 8 adolescents (11/8 with proven/probable CNS-IMD) were included. Seven of the patients died and 12/19 children survived (63%): seven without major neurological sequelae and five with major neurological sequelae. Multifocal ring enhancement and diffusion restriction were the most common brain MRI changes. Diffusion restriction was mostly seen at the core of the lesion. No patient with disease limited to one lobe died. Perivascular microbleeding seen on susceptibility weighted imaging (SWI) and/or gradient-echo/T2* images, as well as infarction, were associated with poor prognosis. Conclusions: The presence of infarction was related to poor outcome. As early microbleeding seems to be associated with poor prognosis, we suggest including SWI in routine diagnostic evaluation of immunocompromised children with suspected CNS-IMD

First Study of Combined Blazar Light Curves with FACT and HAWC

For studying variable sources like blazars, it is crucial to achieve unbiased monitoring, either with dedicated telescopes in pointing mode or survey instruments. At TeV energies, the High Altitude Water Cherenkov (HAWC) observatory monitors approximately two thirds of the sky every day. It uses the water Cherenkov technique, which provides an excellent duty cycle independent of weather and season. The First G-APD Cherenkov Telescope (FACT) monitors a small sample of sources with better sensitivity, using the imaging air Cherenkov technique. Thanks to its camera with silicon-based photosensors, FACT features an excellent detector performance and stability and extends its observations to times with strong moonlight, increasing the duty cycle compared to other imaging air Cherenkov telescopes. As FACT and HAWC have overlapping energy ranges, a joint study can exploit the longer daily coverage given that the observatories' locations are offset by 5.3 hours. Furthermore, the better sensitivity of FACT adds a finer resolution of features on hour-long time scales, while the continuous duty cycle of HAWC ensures evenly sampled long-term coverage. Thus, the two instruments complement each other to provide a more complete picture of blazar variability. In this presentation, the first joint study of light curves from the two instruments will be shown, correlating long-term measurements with daily sampling between air and water Cherenkov telescopes. The presented results focus on the study of the variability of the bright blazars Mrk 421 and Mrk 501 during the last two years featuring various flaring activities.Comment: 6 pages, 2 figures. Contribution to the 6th International Symposium on High Energy Gamma-Ray Astronomy (Gamma2016), Heidelberg, Germany. To be published in the AIP Conference Proceeding

Contribución al conocimiento de Porosagrotis gypaetina (Guen.) (Lep.:Noctuidae)

p.15-22Este trabajo tiene por finalidad brindar una descripcion detallada de los diferentes estados de desarrollo, asi como de los estadios larvales, de Porosagrotis gypaetina (Guen.) y estimar sus principales parametros biologicos. Se trata de una oruga conocida vulgarmente como gusano pardo que frecuenta cultivos de alfalfa, trebol bianco, maiz y girasol y determinadas malezas. Los caracteres considerados para su identificacion fueron, en el huevo: numero y distribucion de costas; en la larva: pigmentacion, distribucion de manchas y cerdas corporales; en la pupa: tamaño, forma y color y caracteristicas del cremaster; y en el adulto: ubicacion y coloracion de maculas y nervaduras alares. La emergencia de imagos alcanzo su maximo en abril y mayo. El periodo embrionario se completo en 22 a 26 dias. Aproximadamente la mitad de las larvas cumplieron su ciclo en 6 estadios y las restantes en 7; la duracion total del periodo larval fue de 134 a 141 dias, sin considerar la forma prepupal e independientemente del numero de estadios. Las orugas permanecieron como prepupas durante la temporada estival (aproximadamente 161 dias). El estado pupal duro 40 a 57 dias. Las observaciones realizadas permiten expresar que, inediante los caracteres descriptos, es factible reconocer la especie a traves no solo de los adultos, sino de sus estados inmaduros. Posee una sola generacion anual; transcurre el inviemo como larva; el daño tipico de corte lo produce a partir del cuarto estadio larval

Stoichiometry of HLA Class II-Invariant Chain Oligomers

BACKGROUND: The HLA gene complex encodes three class II isotypes, DR, DQ, and DP. HLA class II molecules are peptide receptors that present antigens for recognition by T lymphocytes. In antigen presenting cells, the assembly of matched α and β subunits to heterodimers is chaperoned by invariant chain (Ii). Ii forms a homotrimer with three binding sites for class II heterodimers. The current model of class II and Ii structure states that three αβ heterodimers bind to an Ii trimer. METHODOLOGY/PRINCIPAL FINDINGS: [corrected] We have now analyzed the composition and size of the complexes of class II and Ii using epitope tagged class II subunits and density gradient experiments. We show here that class II-Ii oligomers consist of one class II heterodimer associated with one Ii trimer, such that the DR, DQ and DP isotypes are contained within separate complexes with Ii. CONCLUSION/SIGNIFICANCE: We propose a structural model of the class II-Ii oligomer and speculate that the pentameric class II-Ii complex is bent towards the cell membrane, inhibiting the binding of additional class II heterodimers to Ii

Long-term monitoring of bright blazars in the multi-GeV to TeV range with FACT

Blazars like Markarian 421 or Markarian 501 are active galactic nuclei (AGN), with their jets orientated towards the observer. They are among the brightest objects in the very high energy (VHE) gamma ray regime (>100 GeV). Their emitted gamma-ray fluxes are extremely variable, with changing activity levels on timescales between minutes, months, and even years. Several questions are part of the current research, such as the question of the emission regions or the engine of the AGN and the particle acceleration. A dedicated longterm monitoring program is necessary to investigate the properties of blazars in detail. A densely sampled and unbiased light curve allows for observation of both high and low states of the sources, and the combination with multi-wavelength observation could contribute to the answer of several questions mentioned above. FACT (First G-APD Cherenkov Telescope) is the first operational telescope using silicon photomultiplier (SiPM, also known as Geigermode—Avalanche Photo Diode, G-APD) as photon detectors. SiPM have a very homogenous and stable longterm performance, and allow operation even during full moon without any filter, leading to a maximal duty cycle for an Imaging Air Cherenkov Telescope (IACT). Hence, FACT is an ideal device for such a longterm monitoring of bright blazars. A small set of sources (e.g., Markarian 421, Markarian 501, 1ES 1959+650, and 1ES 2344+51.4) is currently being monitored. In this contribution, the FACT telescope and the concept of longterm monitoring of bright blazars will be introduced. The results of the monitoring program will be shown, and the advantages of densely sampled and unbiased light curves will be discussed

DANA Universal Dataflow Analysis for Gate-Level Netlist Reverse Engineering

Reverse engineering of integrated circuits, i.e., understanding the internals of Integrated Circuits (ICs), is required for many benign and malicious applications. Examples of the former are detection of patent infringements, hardware Trojans or Intellectual Property (IP)-theft, as well as interface recovery and defect analysis, while malicious applications include IP-theft and finding insertion points for hardware Trojans. However, regardless of the application, the reverse engineer initially starts with a large unstructured netlist, forming an incomprehensible sea of gates.This work presents DANA, a generic, technology-agnostic, and fully automated dataflow analysis methodology for flattened gate-level netlists. By analyzing the flow of data between individual Flip Flops (FFs), DANA recovers high-level registers. The key idea behind DANA is to combine independent metrics based on structural and control information with a powerful automated architecture. Notably, DANA works without any thresholds, scenario-dependent parameters, or other “magic” values that the user must choose. We evaluate DANA on nine modern hardware designs, ranging from cryptographic co-processors, over CPUs, to the OpenTitan, a stateof- the-art System-on-Chip (SoC), which is maintained by the lowRISC initiative with supporting industry partners like Google and Western Digital. Our results demonstrate almost perfect recovery of registers for all case studies, regardless whether they were synthesized as FPGA or ASIC netlists. Furthermore, we explore two applications for dataflow analysis: we show that the raw output of DANA often already allows to identify crucial components and high-level architecture features and also demonstrate its applicability for detecting simple hardware Trojans.Hence, DANA can be applied universally as the first step when investigating unknown netlists and provides major guidance for human analysts by structuring and condensing the otherwise incomprehensible sea of gates. Our implementation of DANA and all synthesized netlists are available as open source on GitHub

Trained Innate Immunity in Animal Models of Cardiovascular Diseases

Acquisition of immunological memory is an important evolutionary strategy that evolved to protect the host from repetitive challenges from infectious agents. It was believed for a long time that memory formation exclusively occurs in the adaptive part of the immune system with the formation of highly specific memory T cells and B cells. In the past 10–15 years, it has become clear that innate immune cells, such as monocytes, natural killer cells, or neutrophil granulocytes, also have the ability to generate some kind of memory. After the exposure of innate immune cells to certain stimuli, these cells develop an enhanced secondary response with increased cytokine secretion even after an encounter with an unrelated stimulus. This phenomenon has been termed trained innate immunity (TI) and is associated with epigenetic modifications (histone methylation, acetylation) and metabolic alterations (elevated glycolysis, lactate production). TI has been observed in tissue-resident or circulating immune cells but also in bone marrow progenitors. Risk-factors for cardiovascular diseases (CVDs) which are associated with low-grade inflammation, such as hyperglycemia, obesity, or high salt, can also induce TI with a profound impact on the development and progression of CVDs. In this review, we briefly describe basic mechanisms of TI and summarize animal studies which specifically focus on TI in the context of CVDs