The low-virulent African swine fever virus (ASFV/NH/P68) induces enhanced expression and production of relevant regulatory cytokines (IFNα, TNFα and IL12p40) on porcine macrophages in comparison to the highly virulent ASFV/L60

The impact of infection by the low-virulent ASFV/NH/P68 (NHV) and the highly virulent ASFV/L60 (L60) isolates on porcine macrophages was assessed through the quantification of IFNα, TNFα, IL12p40, TGFβ and ASFV genes by real-time PCR at 2, 4 and 6 h post-infection. Increased IFNα, TNFα and IL12p40 expression was found in infection with NHV, in which expression of TGFβ was lower than in infection with L60. Principal component analysis showed a positive interaction of cytokines involved in cellular immune mechanisms, namely IFNα and IL12p40 in the NHV infection. Quantification by ELISA confirmed higher production of IFNα, TNFα and IL12p40 in the NHV-infected macrophages. Overall, our studies reinforce and clarify the effect of the NHV infection by targeting cellular and cellular-based immune responses relevant for pig survival against ASFV infection

User's guide for FINDIF at SACLANTCEN

FINDIF solves the elastic wave equation for a line source in two dimensions by the finite difference method (Virieux,1986; Stephen,1988b). The solution is carried out in the time domain for either pulse or CW sources. Arbitrary distributions of compressional velocity, shear velocity and density (including fluids and solids) can be defined on the finite difference grid. All compressional waves, shear waves, interface waves and evanescent waves are included as are all conversions between wave types. Multiple forward and backward scattering is automatically treatedThis work was cared out under ONR Contract #N00014-89-J-1012 and under ONR Grant #NOOOI490J154

Thermal Characterization of Polyurea Cross-linked Silica Aerogel-RTV 655 for Cryogenic Tank Applications

Thermal conductivities for different volume percentages of polyurea cross-linked silica aerogel to RTV 655 were experimentally determined. Silica aerogel is an ideal insulator with a makeup of up to 99% air. Polyurea cross-linked silica aerogel, PCSA, is a much stronger material while retaining the high thermal insulation properties of this group of materials. RTV 655 can be shaped into any geometry and is stable under extreme temperature conditions. RTV 655 is a space qualified polymer and the compound material made up of aerogel-RTV 655 is being considered here for use as a cryogenic propellant tank material for long duration space missions. Several samples were made to test thermal properties of different volume percentages in an effort to assess the thermal conductivity of the combination of polyurea cross-linked silica aerogel and RTV 655. Thermal conductivity results are presented comparing small aerogel discs embedded in RTV 655, a homogeneous compound consisting of micro scale aerogel particles embedded in RTV 655, large irregular geometry aerogels in RTV 655 and monolithic aerogel blocks embedded in RTV 655

Random amplified polymorphic DNA-PCR and ERIC PCR analysis on Vibrio parahaemolyticus isolated from cockles in Padang, Indonesia

In this study, RAPD-PCR and ERIC-PCR were used to study the epidemiology of V. parahaemolyticus isolated from cockles in Padang, Indonesia. The Gold Oligo OPAR3 primer produced bands ranged from 1-8 with sizes from 0.2 – 5.0 kb and the Gold Oligo OPAR8 primer produced 1-7 bands with sizes 0.7 – 1.5 kb. Both primers produced twenty five RAPD patterns with a few isolates failed to produce any products. Based on phylogenetic dendrogram, all the isolates can be divided into 6 major clusters with similarity between 0 to 52%. For the ERIC primer, it produced bands ranged from 3-15 with sizes from 0.1 – 5.0 kb and twenty seven different ERIC patterns. Construction of the phylogenetic dendogram showed the isolates can be divided into 4 major clusters with similarity between 56 to 86%. The high diversity of both processes may be due to the multiple contamination sources of V. parahaemolyticus

Functional characterization of unassigned african swine fever virus proteins putatively involved in transcription and replication towards an efficient vaccine design

Tese de Doutoramento em Ciências Veterinárias na especialidade de Ciências Biológicas e BiomédicasAfrican swine fever (ASF) is an infectious disease of domestic pigs and wild boars with mortality rates reaching up to 100% and is endemic in most of the Sub-Saharan countries. In 2007 it was introduced in Georgia and spread to neighbouring countries, reaching the Russian Federation, several European countries and, more recently, China and Vietnam (February 2019). Currently, there is neither a vaccine nor a treatment against ASF and the control of the disease depends strictly on sanitary measures, including stamping out and trade bans of animals and pork products leading to devastating socio-economic losses to affected countries. The etiologic agent of the disease is African swine fever virus (ASFV), a large (approx. 190 nm) double-stranded DNA (170 to 193 kbp) enveloped virus. ASFV genome encloses more than 150 open reading frames (ORFs) and to this date most of them lack any known or predictable function. ASFV is quite independent from cellular machinery encoding enzymes required for replication, transcription and virion assembly, including the putative I215L E2 Ubiquitin-conjugating enzyme, QP509L, Q706L RNA Helicases and the P1192R type II topoisomerase. The E2 ubiquitin-conjugating enzymes are part of the essential cellular post-transcriptional regulation ubiquitin-proteasome pathway. In this study, the pI215L binding activity was characterized as being mono and poly-ubiquitinated in the Cys85 at different temperatures and pH values. Moreover, I215L gene is transcribed from 2 hours post infection (hpi), and immunoblot analysis confirmed that pI215L is expressed from 4 hpi being detected all over the cell specially in the viral factories from 8 hpi. Downregulation assays by siRNA suggested that pI215L plays a critical role in the transcription of late viral genes and in viral DNA replication. RNA helicases are described as essential for infections, modulating RNA-RNA and RNA-protein interactions, gene expression, viral egress and host antiviral responses. In the present work, we found that QP509L, Q706L are conserved between ASFV virulent and non-virulent isolates. Furthermore, ASFV-QP509L and Q706L are actively transcribed from 2 hours post infection, and both proteins are localized in the viral factories at 12 hours post infection. However, QP509L was also detected in the cell nucleus. Transcript downregulation uncovered the essential role of these proteins during viral cycle progression, in particular for the late transcription. Type II topoisomerases are involved in resolving DNA tangles and supercoils by cutting the duplex and allowing the DNA replication to proceed. In this study, we report that P1192R is actively transcribed throughout infection, being detected from 2 hpi and reaching a maximum concentration around 16 hpi. P1192R knockdown experiments revealed its critical role for viral infection, given by a reduction in viral transcripts, cytopathic effect, the number of viral factories per cell, and virus yields. We also demonstrated that enrofloxacin exposure during the late phase of infection induces viral genomes fragmentation, whereas, when added at early phase of infection completely abolishes replication. The data obtained from I215L, QP509L. Q706L and P1192R characterization studies opens new venues to the rational design of a mutant virus lacking these genes, and also points new pathways to be targeted by antiviral drugs.RESUMO - Caracterização funcional de proteínas do vírus da peste suína Africana putativamente envolvidas na transcrição e replicação com o intuito de desenvolvimento de uma vacina. - A peste suína africana é uma doença viral infeciosa que afeta os suínos domésticos e os selvagens, com taxas de mortalidade perto dos 100%, originando perdas económicas elevadas nos países afetados. A doença é endémica na maioria dos países subsaarianos, e desde 2007, assistiu-se uma expansão nos países Europeus, incluindo membros da União Europeia, e mais recentemente, na China e Vietname. Atualmente não existe vacina ou tratamento para esta infeção e o controlo da doença baseia-se no diagnóstico rápido, na eliminação compulsiva dos suínos e no bloqueio ao comércio de suínos e produtos derivados. O agente etiológico é o vírus da peste suína africana (VPSA), um vírus composto por uma molécula de ADN de cadeia dupla (170 to 193 kbp) contendo mais de 150 grelhas de leitura. Algumas destas estão devidamente caracterizadas codificando para proteínas estruturais ou regulatórias, contudo, a grande maioria foi identificada por homologia de sequência com outros vírus não se conhecendo, até à data, qual a sua função durante a infeção. Apesar dos inúmeros esforços ao longo dos anos, a complexidade viral, a falta de conhecimento sobre muitos dos aspetos da biologia do vírus e das suas interações com o hospedeiro invalidaram a obtenção de uma vacina segura e eficaz. Por um lado, as abordagens clássicas embora promissoras não garantem proteção contra estirpes heterólogas, enquanto a produção de vacinas de ADN ou proteína, mesmo com adjuvantes, não induzem imunidade contra uma segunda infeção. No entanto, a identificação de suínos previamente infetados e que resistem a novas infeções reforça a ideia da possibilidade de se obter uma imunidade protetora. Dadas as circunstâncias atuais de expansão da doença, estudos recentes apontam a necessidade de se aprofundar o conhecimento sobre os aspetos da biologia do VPSA com vista a identificação de novas estratégias para o desenvolvimento racional de vacinas ou de identificação de novos alvos para o uso de fármacos com vista a controlar a infeção. Neste contexto, os estudos apresentados neste trabalho caracterizam a I215L, QP509L, Q706L e P1192R, identificadas inicialmente, por homologia de sequência com outras proteínas tipicamente envolvidas na replicação e transcrição de outros vírus. A I215L foi identificada por partilhar identidade com as enzimas E2 de conjugação da ubiquitina. Estas enzimas pertencem a uma cadeia de sinalização do sistema de regulação pós-transcricional ubiquitina-proteossoma. Os estudos realizados revelaram que a pI215L tem a capacidade de receber uma ou duas ubiquitinas (mono e di-ubiquitinada) no resíduo Cisteína-85, a diferentes temperaturas e valores de pH, evidenciando a sua plasticidade em participar em diferentes fases da infeção quer no hospedeiro quer no vetor. Além disto, o gene é transcrito precocemente (2 horas após infecção, hpi) e a proteína expressa desde as 4h, sugerindo que esta deverá ser necessária desde o início da infecção. Paralelamente, os nossos estudos por imunofluorescência revelaram uma distribuição da pI215L por toda a célula, e em especial, nas fábricas virais, sugerindo um papel ativo na regulação de vários processos, incluindo replicação de ADN e da transcrição. Os ensaios de ARN de interferência (siRNA) contra o I215L demonstraram um papel essencial desta proteína durante a infeção, originando uma redução dos transcritos tardios, do número de genomas (-63 a -68%) e na libertação de partículas infeciosas (até -94%). [...]N/

Preliminary identification of Envelope Protein VP32 and Collagen-like protein CLP of White Spot Syndrome Virus

对虾白斑综合症病毒(whitespotsyndromevirus，WSSV)是一种具有囊膜、无包涵体的杆状型双链环状DNA病毒，Nimaviridae属Whispovirus种中唯一的物种，也是严重危害对虾养殖业的最主要病原。 目前我们的研究主要围绕WSSV的蛋白功能开展研究。这是因为病毒蛋白参与病毒的形态构成、对宿主细胞的入侵、增殖等一系列活动，并在其中扮演了重要角色。 本论文研究分为以下两个部分： （1）质谱发现病毒膜蛋白VP32是ORFwsv198的编码产物。我们构建了融合表达载体pGEX-2Th-vp32和pMAL-vp32并分别在E.coliBL-21菌株中表达纯化了GST-V...White Spot Syndrome Virus (WSSV) is a large, rod-shaped, enveloped double-stranded DNA virus. The WSSV has been classified to be the sole species of Whispovirus in genus of Nimaviridae, and the main pathogen harming shrimp industry. At present, our research mainly focuses on the main proteins of WSSV. These proteins play important roles in the formation construction, invasion, proliferation, etc...学位：理学硕士院系专业：生命科学学院生物学系_细胞生物学学号：2012005130206

Elastodynamics of a soft strip subject to a large deformation

To produce sounds, we adjust the tension of our vocal cords to shape their properties and control the pitch. This efficient mechanism offers inspiration for designing reconfigurable materials and adaptable soft robots. However, understanding how flexible structures respond to a significant static strain is not straightforward. This complexity also limits the precision of medical imaging when applied to tensioned organs like muscles, tendons, ligaments and blood vessels among others. In this article, we experimentally and theoretically explore the dynamics of a soft strip subject to a substantial static extension, up to 180\%. Our observations reveal a few intriguing effects, such as the resilience of certain vibrational modes to a static deformation. These observations are supported by a model based on the incremental displacement theory. This has promising practical implications for characterizing soft materials but also for scenarios where external actions can be used to tune properties

Interkulturelle Kommunikation : Kosten und Nutzen einer Lingua franca

Die Arbeit setzt sich mit der Bedeutung der Lingua franca in schriftlicher, online geführter Kommunikation multikultureller Teams auseinander. Dabei wird untersucht, ob sich L2-Sprecher in einer online geführten Lingua-franca-Kommunikation weniger – sowie syntaktisch und inhaltlich vereinfacht – äußern. Im Blickfeld stehen auch die Fragen, ob L1-Sprecher in einer online geführten Lingua-franca-Kommunikation vermehrt kommunikative Führungsrollen übernehmen und ihre Ideen dadurch eher durchsetzen können und ob diskursive Kommunikationskompetenzen den L2-Sprechern ebenfalls Leitungsfunktionen in einer Lingua-franca-Kommunikation ermöglichen. Nach einem einführenden historischen Rückblick auf die Entwicklung des Forschungsbereichs der Interkulturellen Kommunikation wird mithilfe des methodenintegrierenden Ansatzes der Triangulation zunächst das Kommunikationsverhalten multikultureller online kommunizierender Mitarbeiter der pharmazeutischen Firma Interkult analysiert. Im Anschluss daran werden die Ergebnisse in einem kontrollierbaren Rahmen am Seminar für Deutsch als Fremdsprachenphilologie der Universität Heidelberg anhand einer empirischen Untersuchung erweitert und überprüft: 40 Studierende der Sprachgruppen Deutsch, Englisch, Französisch und Polnisch haben sich an einer Aufgabenstellung orientiert, für die sie auf einem Onlineforum über einen Zeitraum von vier Wochen sowohl in ihrer jeweiligen L1 als auch in der Lingua franca Deutsch Lösungsmöglichkeiten diskutieren sollten. Anhand der Ergebnisse werden die anfangs genannten Fragen hinsichtlich der Rolle der Lingua franca in schriftlicher, online geführter Kommunikation multikultureller Teams bearbeitet

Evaluation der perzeptiven Testlistenäquivalenz des Freiburger Einsilbertests mit Normalhörenden aus dem südostdeutschen Sprachraum

Ausgangspunkt für diese Dissertation ist eine widersprüchliche Studienlage, denn der Freiburger Einsilbertest wird seit Jahren immer wieder kritisiert. Bemängelt werden u.a. der Bekanntheitsgrad und die Aktualität der Testwörter, die phonemische Ausgewogenheit, die Test-Retest-Kontinuität und die perzeptive Äquivalenz der Testlistenschwierigkeiten. Die Zielsetzung dieser Arbeit ist es, den Freiburger Einsilbertest insbesondere auf die perzeptive Testlistenäquivalenz erneut zu überprüfen und eine Empfehlung für die Verwendung der Testlisten auszuprechen. Material und Methoden Mit einer Auswahl von 40 Probanden (31 w, 9 m), otologisch unauffällig / normalhörend, ⌀ 22,6 Jahre, Herkunft Südostdeutschland, wurde der Freiburger Einsilbertest an jedem Probanden mit allen 20 Testlisten bei 4 unterschiedlichen Schallpegeln (50 dB, 35 dB, 30 dB und 20 dB) getestet. Die sprachaudiometrische Messung erfolgte monaural und mit dem Kopfhörer Sennheiser HDA200, es folgte eine statistische Auswertung. Die Arbeit beinhaltet 3 Hypothesen: 1. Hypothese: Es finden sich keine Abweichungen der listenspezifischen mittleren Sprachverständlichkeit zu einer durchschnittlichen Schwierigkeit aus allen Testlisten durchgängig bei allen Sprachpegeln. 2. Hypothese: Es gibt keine Unterschiede der Testlistenschwierigkeit bei einem paarweisen Vergleich zweier Testlisten. Dies wurde bisher noch nicht berücksichtig, hat aber große Auswirkungen auf den Praxisalltag, da ein direkter Ergebnisvergleich von zwei Testlisten in der Praxis am Patient wohl am häufigsten ist, z.B. bei der Vergabe und Überprüfung von Hörgeräten. 3. Hypothese: Es gibt keinen Einfluss der regional spezifischen Sprachentwicklung der Probanden und bei einem Vergleich mit der Arbeit von Baljic et al. (2016) findet sich kein Unterschied bei den testlistenspezifischen Schwierigkeiten der Testlisten. Ergebnisse und Diskussion Alle drei Hypothesen werden abgelehnt. (1) Es wurden zwar nicht durchgängig bei allen Schallpegeln Abweichungen von einer mittleren Testlistenschwierigkeit gefunden, jedoch fiel die Testliste 16 bei 50 dB als schwieriger und die Testliste 15 bei 30 dB als leichter auf, trotz konservativer statistischer Auswertung. (2) Bei den paarweisen Vergleichen wurden bei mehreren Testlisten signifikante Abweichungen festgestellt, am stärksten betroffen waren die Testlisten 6,12,14,15,16,19, und 20. Die Bedeutung der paarweisen Vergleiche wurde bisher unterschätzt und sollte in Zukunft stärker berücksichtig werden. (3) Bei der Untersuchung eines Einflusses der regional spezifischen Sprachentwicklung wurde das Verfahren der Vergleichsarbeit von Baljic et al. (2016) verwendet. Dabei zeigten sich bei den auffälligen Testlisten sowohl Unterschiede, als auch Übereinstimmungen. Dies zeigt, dass ein regionaler Einfluss nicht ausgeschlossen werden kann, aber auch nicht der alleinige Faktor ist. Fazit Der Freiburger Sprachverständlichkeitstest hat nach wie vor seine Berechtigung bei der Untersuchung der Grundverständlichkeit in Ruhe als kontextfreie Basis. Aufgrund der widersprüchlichen Studie sollte jedoch in Erwägung gezogen werden, den Test nach beinahe über 50 Jahren zu aktualisieren, oder, soweit möglich, auf Alternativen wie z.B. den Reimtest nach Wallenberg und Kollmeier oder Satztests auszuweichen. Nach einem Abgleich verschiedener Studien können die Testlisten 3,4,7,9 und 10 am meisten für die Anwendung empfohlen werden, da sie bisher in allen Studien am wenigsten aufgefallen waren. Am wenigsten zu empfehlen sind dagegen die Testlisten 12 und 15

Detection of Gamma-ray Emission from the Eta-Carinae Region

We present the results of extensive observations by the gamma-ray AGILE satellite of the Galactic region hosting the Carina nebula and the remarkable colliding wind binary Eta Carinae (Eta Car) during the period 2007 July to 2009 January. We detect a gamma-ray source (1AGL J1043-5931) consistent with the position of Eta Car. If 1AGL J1043-5931 is associated with the Eta Car system our data provide the long sought first detection above 100 MeV of a colliding wind binary. The average gamma-ray flux above 100 MeV and integrated over the pre-periastron period 2007 July to 2008 October is F = (37 +/- 5) x 10-8 ph cm-2 s-1 corresponding to an average gamma-ray luminosity of L = 3.4 x 10^34 erg s-1 for a distance of 2.3 kpc. We also report a 2-day gamma-ray flaring episode of 1AGL J1043-5931 on 2008 Oct. 11-13 possibly related to a transient acceleration and radiation episode of the strongly variable shock in the system.Comment: 5 pages, 4 figure, accepted for publication in ApJ Letter