EFFICIENT PROPAGATION OF ARCHETYPE JC POLYOMAVIRUS IN COS-7 CELLS: EVALUATION OF REARRANGEMENTS WITHIN NCCR STRUCTURAL ORGANIZATION DURING TRANSFECTION.

John Cunningham virus (JCPyV) is an ubiqui-tous human pathogen that causes disease in immunocom-promised patients. The JCPyV genome is composed of an early region and a late region, which are physically sepa-rated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robust-ness. Prior to this study, it was demonstrated that JCPyV archetype DNA replicates in COS-7 simian kidney cells expressing SV40 TAg and COS-7 cells expressing HIV-1 Tat. Based on these observations, the present study was conducted to reproduce an in vitro model in COS-7 cells transfected with the JCPyV archetype strain in order to study JCPyV DNA replication and analyze NCCR rear-rangements during the viral life cycle. The efficiency of JCPyV replication was evaluated by quantitative PCR (Q-PCR) and by hemagglutination (HA) assay after trans-fection. In parallel, sequence analysis of JCPyV NCCR was performed. JCPyV efficiently replicated in kidney-derived COS-7 cells, as demonstrated by a progressive increase in viral load and virion particle production after transfection. The archetypal structure of NCCR was maintained during the viral cycle, but two characteristic point mutations were detected 28 days after transfection. This model is a useful tool for analyzing NCCR rearrangements during in vitroreplication in cells that are sites of viral persistence, such as tubular epithelial cells of the kidne

Yersinia enterocolitica in Italy. A case of septicemia and abdominal aortic aneurysm infection

We report a case of Yersinia enterocolitica septicemia in a 63-year-old patient admitted to the Vascular Surgery Department of Umberto I Hospital (Rome, Italy) for an abdominal aortic aneurysm. The microorganism, recovered from both peripheral blood cultures and aneurysmatic aortic wall specimens, was identified as Y. enterocolitica using matrix-assisted laser desorption ionization-time of flight analysis (MALDI-TOF MS) and 16S rDNA gene sequencing. The isolate responsible for septicemia belonged to the O:9 serotype (biogroup 2). A genetic screening of the isolate made it possible to detect the presence of both the yst and ail genes, encoding a heat-stable enterotoxin and a protein involved in invasion/adherence and serum resistance, respectively. Our case contributes in enriching epidemiological data concerning Y. enterocolitica infections, which might represent severe complications in patients suffering from cardiovascular diseases. Moreover, this study, together with the others, should be regarded as valuable and useful tools for monitoring the rate of infections worldwide

High frequency of JCV DNA detection in prostate cancer tissues

BACKGROUND: Prostate cancer (PC) represents the most frequently diagnosed cancer in men. Exposure to infectious agents has been considered to induce prostatic inflammation and cancerous transformation. Controversial data exist concerning the role of the human polyomaviruses BK (BKV) and JC (JCV) in PC etiology. Therefore, a possible association between these polyomaviruses and PC was investigated. MATERIALS AND METHODS: Urine, blood and fresh prostatic tissue specimens were collected from 26 patients with PC. The presence of BKV and JCV, the possible non-coding control region (NCCR) variations and the genotyping analysis of viral protein 1 (VP1) of both viruses were assessed. RESULTS: Data showed a preferential viral re-activation in the urinary compartment and a statistically significant prevalence of JC viruria and of BKV in PC tissues. A BKV DDP-like NCCR sequence was isolated in two patients, whereas JCV NCCR was consistently of an archetypal structural organization. A prevalence of the European genotypes was observed for both viruses. CONCLUSION: Our data demonstrated the presence of JCV DNA in 14/24 (58.3%) cancerous prostatic tissue specimens, confirming the results obtained in a previous study, in which JCV has been defined as common inhabitant of the prostate, and opening the discussion about its potential role in PC

O currículo da educação de jovens e adultos e o ensino de ciências: um olhar sobre a cultura

Atuar na investigação sobre a Educação de Jovens e Adultos (EJA) não delimita apenas a reflexão sobre uma faixa etária diferenciada de alunos, mas remete, dentre outros elementos, ao pensar e agir junto a um grupo com especificidades culturais. Alguns currículos brasileiros direcionados ao ensino e aprendizagem na disciplina de Ciências na EJA vêm aumentando a atenção quanto à heterogeneidade cultural, como resultado do maior conhecimento acerca dos sujeitos dessa aprendizagem. No presente artigo expomos resultados de uma investigação qualitativa, que considerou como instrumentos a pesquisa bibliográfica e documental, sobre como a heterogeneidade cultural é contemplada nas diretrizes ou orientações curriculares nacionais (do Brasil), estaduais (do Paraná/Brasil) e municipais (de Cascavel/Paraná/Brasil), especialmente nos direcionamentos para a disciplina de Ciências. Os documentos norteadores da EJA, como pareceres, resoluções, propostas curriculares e diretrizes curriculares, foram estudados tendo por abordagem metodológica principal a Análise de Conteúdo. Tal ação pretende contribuir para a implementação de políticas e práticas que almejem a ação educativa pautada no respeito à cultura do aluno e da imersão na cultura científic

Acciones de un equipo multidisciplinar como experiencia en la educación formal.

El presente artículo presenta los objetivos y actividades de un proyecto con enfoque en la diversidad, desarrollado en una escuela pública estatal, hecho desde el año 2010, con reediciones anuales. El proyecto que recibió el nombre de Projeto de Intervenção da Equipe Multidisciplinar desarrolla acciones de reflexión, formación y enfrentamiento a la discriminación y al prejuicio, en todas sus formas, celando por el respeto a la diversidad presente en el espacio escolar. Las actividades propuestas son organizadas por el Equipo Multidisciplinar, integrado por profesores y otros profesionales de educación del “Colegio Estadual Humberto de Alencar Castelo Branco – Ensino Médio e Normal” en el municipio de Santa Helena-Paraná, Brasil. Las actividades realizadas por el Equipo Multidisciplinar son evaluadas por la comunidad escolar por medio de la participación en las acciones a ser desarrolladas y por el análisis de los registros hechos por el Equipo, que son registradas en actas y, enseguida archivadas. Los resultados obtenidos en estos años de efectivo trabajo de la propuesta apuntan para actitudes de respeto a la diversidad cultural existente en el colegio, así como la valoración de las diferentes culturas existentes en la socieda

NASA Trapezoidal Wing Simulation Using Stress-w and One- and Two-Equation Turbulence Models

The Wilcox 2006 stress-omega model (also referred to as WilcoxRSM-w2006) has been implemented in the NASA Langley code CFL3D and used to study a variety of 2-D and 3-D configurations. It predicted a variety of basic cases reasonably well, including secondary flow in a supersonic rectangular duct. One- and two-equation turbulence models that employ the Boussinesq constitutive relation were unable to predict this secondary flow accurately because it is driven by normal turbulent stress differences. For the NASA trapezoidal wing at high angles of attack, the WilcoxRSM-w2006 model predicted lower maximum lift than experiment, similar to results of a two-equation model

JC virus-DNA detection is associated with CD8 fffector accumulation in peripheral blood of patients with multiple sclerosis under natalizumab treatment, independently from JC virus serostatus

Although natalizumab (anti-α4 integrin) represents an effective therapy for relapsing remitting multiple sclerosis (RRMS), it is associated with an increased risk of developing progressive multifocal leukoencephalopathy (PML), caused by the polyomavirus JC (JCV). The aim of this study was to explore natalizumab-induced phenotypic changes in peripheral blood T-lymphocytes and their relationship with JCV reactivation. Forty-four patients affected by RRMS were enrolled. Blood and urine samples were classified according to natalizumab infusion number: 0 (N0), 1-12 (N12), 13-24 (N24), 25-36 (N36), and over 36 (N > 36) infusions. JCV-DNA was detected in plasma and urine. T-lymphocyte phenotype was evaluated with flow cytometry. JCV serostatus was assessed. Ten healthy donors (HD), whose ages and sexes matched with the RRMS patients of the N0 group, were enrolled. CD8 effector (CD8 E) percentages were increased in natalizumab treated patients with detectable JCV-DNA in plasma or urine compared to JCV-DNA negative patients (JCV-) (p < 0.01 and p < 0.001, resp.). Patients with CD8 E percentages above 10.4% tended to show detectable JCV-DNA in plasma and/or urine (ROC curve p = 0.001). The CD8 E was increased when JCV-DNA was detectable in plasma or urine, independently from JCV serology, for N12 and N24 groups (p < 0.01). As long as PML can affect RRMS patients under natalizumab treatment with a negative JCV serology, the assessment of CD8 E could help in the evaluation of JCV reactivation

MALDI-TOF MS Versus VITEK®2: Comparison of Systems for the Identification of Microorganisms Responsible for Bacteremia

We evaluated the reliability and accuracy of the combined use of MALDI-TOF MS and classical ID VITEK2 to identify monomicrobial infection in blood culture bottles. In total, 70 consecutive positive blood cultures were included in this study. Positive blood culture bottles were subjected to Gram staining and subcultured on solid media. Isolates grown from such culture media were used for classical ID using VITEK2 system. In parallel, an aliquot was subjected to a lysing-centrifugation method and used for the identification with the MALDI-TOF system. Results evidenced the correct genus and species identification of 91.4 % of microorganisms responsible for bacteremia with an agreement to the species and the genus level. If compared with the standard method VITEK2, our simple and cost-effective sample preparation method would be very useful for rapid identification of microorganisms using blood culture bottles. In fact, the direct method showed rapid and reliable results, especially for the gram-negative group

A decentralized approach for enabling advanced ancillary services through distributed energy sources

This thesis reports the results carried out by the author during the three-year activities of the XXXV cycle of the Ph.D. course in Electrical and Information Engineering at Politecnico di Bari. The main goal of this work was to implement a decentralized approach to enable the provision of Ancillary Services (AS) by means of Distributed Energy Resources (DERs). As a first step, an analysis of Regulations, Directives, and Network Codes at the national and European level was carried out to define the future framework of the electricity market. In addition, papers and technical reports concerning eight European projects, such as SmarNet, CoordiNet, FLEXCoop, OSMOSE, inteGRIDy, eDREAM, ADDRESS, and evolvDSO have been analyzed with the aim to define the state of the art about the provision of AS by means of DERs. In particular, a set of coordination schemes was identified from those proposed in the literature to coordinate the use of DERs among Transmission System Operators (TSOs) and Distribution System Operators (DSOs). These coordination schemes are classified into centralized and decentralized architectures, depending on the possibility of System Operators of managing their respective networks. In addition, the main methodologies proposed in the literature to perform optimal dispatch of flexibility resources in a TSO-DSO coordination framework, taking technical and grid constraints into account, have been identified and presented. In order to enable DERs in AS provision and coordinate system operators in their usage, a few methodologies suitable for decentralized architectures were developed and presented in this work. In particular, a new algorithm based on a three-phase optimal power flow routine for mapping the flexibility area at the point of interconnection (POI) between transmission and distribution grids was provided. Furthermore, two alternative methodologies to aggregate flexibility resources located in distribution networks interfaced with the TN in multiple POIs were described. In addition, a Benders decomposition algorithm able to optimize TN and DN flexibility resources for congestion management in a few decentralized coordination schemes was also implemented. The proposed methodologies were validated by means of simulation tests conducted on a power system including transmission and distribution grids. The last part of this thesis investigated how innovative grid services, such as Synthetic Inertia (SI) and Fast Frequency Response (FFR), may be also provided through fast control dispatchable resources located at the distribution grid level. With this aim, a low-cost controller for end-user applications able to generate a SI law for dispatchable DERs was developed and tested through Power Hardware-in-the-Loop (PHIL) tests. Similarly, PHIL tests were also carried out to investigate the capability of LED lamps of public lighting systems to be controlled with the aim to provide SI and FFR actions. Experimental results validated the proposed decentralized approaches