Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques

Background: Tomato brown rugose fruit virus (ToBRFV) is a highly infectious tobamovirus that causes severe disease in tomato (Solanum lycopersicum L.) crops. In Italy, the first ToBRFV outbreak occurred in 2018 in several provinces of the Sicily region. ToBRFV outbreak represents a serious threat for tomato crops in Italy and the Mediterranean Basin. Methods: Molecular and biological characterisation of the Sicilian ToBRFV ToB-SIC01/19 isolate was performed, and a sensitive and specific Real-time RT-PCR TaqMan minor groove binder probe method was developed to detect ToBRFV in infected plants and seeds. Moreover, four different sample preparation procedures (immunocapture, total RNA extraction, direct crude extract and leaf-disk crude extract) were evaluated. Results: The Sicilian isolate ToB-SIC01/19 (6,391 nt) showed a strong sequence identity with the isolates TBRFV-P12-3H and TBRFV-P12-3G from Germany, Tom1-Jo from Jordan and TBRFV-IL from Israel. The ToB-SIC01/19 isolate was successfully transmitted by mechanical inoculations in S. lycopersicum L. and Capsicum annuum L., but no transmission occurred in S. melongena L. The developed real-time RT-PCR, based on the use of a primer set designed on conserved sequences in the open reading frames3, enabled a reliable quantitative detection. This method allowed clear discrimination of ToBRFV from other viruses belonging to the genus Tobamovirus, minimising false-negative results. Using immunocapture and total RNA extraction procedures, the real-time RT-PCR and end-point RT-PCR gave the same comparable results. Using direct crude extracts and leaf-disk crude extracts, the end-point RT-PCR was unable to provide a reliable result. This developed highly specific and sensitive real-time RT-PCR assay will be a particularly valuable tool for early ToBRFV diagnosis, optimising procedures in terms of costs and time

Leveraging on Digital Signage Networks to Bring Connectivity IOT Devices

The number of Internet-connected devices exceeds the world’s population by more than three times and this figure is expected to be doubled within the next five years. The Internet of Things is a concept that describes this trend and outlines certain aspects of design and functionality that new devices should incorporate for a successful integration into the Internet. In this respect, Digital Signage networks traditionally used for audiovisual media, accomplish many of the characteristics of the Internet of Things devices: interoperability, mobility, scalability and ubiquity, both in terms of access and control of devices and regarding the information they generate. This paper raises the power to employ a proposed Digital Signage network as a substrate to connect other types of devices that can benefit from the advantages of this kind of networks. For that aim, the main problems for this integration are discussed, mainly those related to the bidirectional tunneling scheme used in the proposed Digital Signage solution. The effects of this tunneling approach are analyzed in scenarios with bandwidth constraints, and different solutions are proposed. Tunneling performance in mobility is improved, to increase the amount of Internet of Things devices and applications that can benefit from this type of network. El número de dispositivos conectados a Internet supera actualmente a la población mundial por más de tres veces y se espera que esta cifra se duplique en los próximos cinco años. El Internet de las Cosas es un concepto que describe esta tendencia y perfila ciertos aspectos de diseño y funcionalidad que los nuevos dispositivos deben incorporar para lograr una integración exitosa en Internet. En este sentido, las redes digital signage utilizadas tradicionalmente para los medios de comunicación audiovisual cumplen muchas de las características requeridas en el contexto del Internet de las Cosas: interoperabilidad, movilidad, escalabilidad y ubicuidad; relativas tanto al acceso y control de dispositivos como a la información que estos generan. En este trabajo se plantea el poder de emplear la red digital signage propuesta como sustrato para poder conectar otros tipos de dispositivos para que así puedan aprovechar las ventajas de estas redes. Para ese fin, se discuten los principales problemas existentes en esta integración, prestando especial atención al esquema de túnel bidireccional utilizado en la solución digital signage propuesta. Los efectos de este enfoque de tunelación se analizan en escenarios con limitaciones de ancho de banda y se proponen diferentes soluciones. Con ello se consigue mejorar el rendimiento del túnel en movilidad, facilitando la integración de más dispositivos al Internet de las Cosas al permitir que puedan integrarse en este tipo de redes

CD8⁺ T Cell Activation Leads to Constitutive Formation of Liver Tissue-Resident Memory T Cells that Seed a Large and Flexible Niche in the Liver

Liver tissue-resident memory T (Trm) cells migrate throughout the sinusoids and are capable of protecting against malaria sporozoite challenge. To gain an understanding of liver Trm cell development, we examined various conditions for their formation. Although liver Trm cells were found in naive mice, their presence was dictated by antigen specificity and required IL-15. Liver Trm cells also formed after adoptive transfer of in vitro-activated but not naive CD8+ T cells, indicating that activation was essential but that antigen presentation within the liver was not obligatory. These Trm cells patrolled the liver sinusoids with a half-life of 36 days and occupied a large niche that could be added to sequentially without effect on subsequent Trm cell cohorts. Together, our findings indicate that liver Trm cells form as a normal consequence of CD8+ T cell activation during essentially any infection but that inflammatory and antigenic signals preferentially tailor their development. Holz et al. demonstrate that tissue-resident memory T (Trm) cells routinely develop in the liver after T cell activation. Within the liver, IL-15, antigen, and inflammation aid Trm cell formation, but only IL-15 is essential. Newly formed Trm cells do not displace existing populations, demonstrating a flexible liver niche

COLLAGE: a collaborative Learning Design editor based on patterns

CSCL (Computer-Supported Collaborative Learning) constitutes a significant field that has drawn the attention of many researchers and practitioners (Dillenbourg, 2002). This domain is characterized by the coexistence of very different expectations, requirements, knowledge and interests posed by both collaborative learning practitioners and experts in information and communication technologies. In other words, CSCL is an intrinsically interdisciplinary field that implies a need for mutual understanding among the implied stakeholders. This need demands the active participation of all these stakeholders during the whole development cycle of CSCL solutions. Participatory Design (PD) approaches (Muller & Kuhn, 1993) propose a diversity of theories, practices, etc. with the goal of working directly with users and other stakeholders in the design of social systems. That is, PD methodologies define processes where users and developers work together during a certain period of time, while they identify the requirements of an application. In the CSCL case, it has been shown that it is not efficient enough to simply perform the identification and analysis of requirements for the development of CSCL solutions that support effective ways of learning. Collaborative learning practitioners also become active players in the process of customizing technological solutions to their particular needs in every learning situation. PD poses a new requirement that CSCL developers should tackle: how to obtain technological solutions for collaborative learning capable of being particularized/customized by practitioners that usually do not have technological skills

Plasmodium berghei Hsp90 contains a natural immunogenic I-A^b-restricted antigen common to rodent and human Plasmodium species

Thorough understanding of the role of CD4 T cells in immunity can be greatly assisted by the study of responses to defined specificities. This requires knowledge of Plasmodium-derived immunogenic epitopes, of which only a few have been identified, especially for the mouse C57BL/6 background. We recently developed a TCR transgenic mouse line, termed PbT-II, that produces CD4+ T cells specific for an MHC class II (I-Ab)-restricted Plasmodium epitope and is responsive to both sporozoites and blood-stage P. berghei. Here, we identify a peptide within the P. berghei heat shock protein 90 as the cognate epitope recognised by PbT-II cells. We show that C57BL/6 mice infected with P. berghei blood-stage induce an endogenous CD4 T cell response specific for this epitope, indicating cells of similar specificity to PbT-II cells are present in the naïve repertoire. Adoptive transfer of in vitro activated TH1-, or particularly TH2-polarised PbT-II cells improved control of P. berghei parasitemia in C57BL/6 mice and drastically reduced the onset of experimental cerebral malaria. Our results identify a versatile, potentially protective MHC-II restricted epitope useful for exploration of CD4 T cell-mediated immunity and vaccination strategies against malaria

Nuclear medium modification of the F2 structure function

We study the nuclear effects in the electromagnetic structure function F2(x,Q^2) in nuclei in the deep inelastic lepton nucleus scattering process by taking into account Fermi motion, binding, pion and rho meson cloud contributions. Calculations have been done in a local density approximation using relativistic nuclear spectral functions which include nucleon correlations for nuclear matter. The ratios over deuteron structure function are obtained and compared with the recent JLAB results for light nuclei with special attention to the slope of the x distributions. This magnitude shows a non trivial A dependence and it is insensitive to possible normalization uncertainties. The results have also been compared with some of the older experiments using intermediate mass nuclei.Comment: 19 pages, 8 figures. This version matches accepted version to be published in Nuclear Physics

Ianus: an Adpative FPGA Computer

Dedicated machines designed for specific computational algorithms can outperform conventional computers by several orders of magnitude. In this note we describe {\it Ianus}, a new generation FPGA based machine and its basic features: hardware integration and wide reprogrammability. Our goal is to build a machine that can fully exploit the performance potential of new generation FPGA devices. We also plan a software platform which simplifies its programming, in order to extend its intended range of application to a wide class of interesting and computationally demanding problems. The decision to develop a dedicated processor is a complex one, involving careful assessment of its performance lead, during its expected lifetime, over traditional computers, taking into account their performance increase, as predicted by Moore's law. We discuss this point in detail

ACTN3 R577X Polymorphism does not Influence Explosive Leg Muscle Power in Elite Volleyball Players

We examined the association of R577X polymorphism (rs1815739) in the α-actinin-3 (ACTN3) gene with “explosive” leg muscle power performance in a group of male and female elite volleyball players (n=66, 31 men, 35 women) and in a group of non-athletic male and female young adults (n=334, 243 men, 91 women). We assessed power performance by means of the vertical squat and counter-movement jump tests. We also determined whether the genotypic frequencies of the ACTN3 R577X genotypes differed between groups. We did not observe any effect of the ACTN3 R577X polymorphism on study phenotypes in both groups, regardless of gender (all P>0.05). Genotype frequencies were similar between volleyball and control groups (P=0.095). Moreover, we did not find an association between the ACTN3 R577X polymorphism and the likelihood of being an elite volleyball player using the dominant (RR vs RX+XX) and the recessive model (RR+RX vs XX). In summary, these findings suggest that the ACTN3 R577X polymorphism does not influence explosive leg muscle power in elite volleyball players