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Nested PCR to detect and distinguish the sympatric filarial species Onchocerca volvulus, Mansonella ozzardi and Mansonella perstans in the Amazon Region

  • Ta Tang, Thuy-Huong
  • López-Vélez, Rogelio
  • Lanza-Suarez, Marta
  • Shelley, Anthony John
  • Rubio Muñoz, Jose Miguel
  • Luz, Sérgio Luiz Bessa
We present filaria-nested polymerase chain reaction (PCR), which is based on amplification of first internal transcribed spacer rDNA to distinguish three parasitic filarial species (Onchocerca volvulus, Mansonella ozzardi and Mansonella perstans) that can be found in the Amazon Region. Nested PCR-based identifications yielded the same results as those utilizing morphological characters. Nested PCR is highly sensitive and specific and it detects low-level infections in both humans and vectors. No cross-amplifications were observed with various other blood parasites and no false-positive results were obtained with the nested PCR. The method works efficiently with whole-blood, blood-spot and skin biopsy samples. Our method may thus be suitable for assessing the efficacy of filaria control programmes in Amazonia by recording parasite infections in both the human host and the vector. By specifically differentiating the major sympatric species of filaria, this technique could also enhance epidemiological research in the region., Sí

First evidence of a pro-inflammatory response to severe infection with influenza virus H1N1

  • Fernández-de Castro, Isabel
  • Guzmán-Fulgencio, María
  • Garcia-Alvarez, Monica
  • Resino, Salvador
The great majority of infections caused by the pandemic variant of the influenza virus (nvH1N1) are self-limited, but a small percentage of patients develop severe symptoms requiring hospitalization. Bermejo-Martin and colleagues have presented a pilot study describing the differences in the early immune response for patients both mildly and severely infected with nvH1N1. Patients who develop severe symptoms after nvH1N1 infection showed Th1 and Th17 'hypercytokinemia', compared to mildly infected patients and healthy controls. The mediators involved with the Th1 and Th17 profiles are known to be involved in antiviral, pro-inflammatory and autoimmune responses. This is the first work reporting the association of a pro-inflamatory immune response with a severe pandemic infection, although it is likely that more studies are needed to understand the detrimental or beneficial roles these cytokines play in the evolution of mild and severe nvH1N1 infection., This work was supported by grants from Instituto de Salud Carlos III (PI08/0738; UIPY 1467/07) to SR. MG-F is supported by a grant of Instituto de Salud Carlos III (CM09/00031). MG-A is supported by a grant of Instituto de Salud Carlos III (CM08/00101)., Sí

Initial fitness recovery of HIV-1 is associated with quasispecies heterogeneity and can occur without modifications in the consensus sequence

  • Borderia, Antonio V.
  • Lorenzo-Redondo, Ramon
  • Pernas, Maria
  • Casado, Concepcion
  • Alvaro-Cifuentes, Tamara
  • Domingo, Esteban
  • Lopez-Galindez, Luis Cecilio
BACKGROUND: Fitness recovery of HIV-1 "in vitro" was studied using viral clones that had their fitness decreased as a result of plaque-to-plaque passages. PRINCIPAL FINDINGS: After ten large population passages, the viral populations showed an average increase of fitness, although with wide variations among clones. While 5 clones showed significant fitness increases, 3 clones showed increases that were only marginally significant (p<0.1), and 4 clones did not show any change. Fitness recovery was not accompanied by an increase in p24 production, but was associated with an increase in viral titer. Few mutations (an average of 2 mutations per genome) were detected in the consensus nucleotide sequence of the entire genome in all viral populations. Five of the populations did not fix any mutation, and three of them displayed marginally significant fitness increases, illustrating that fitness recovery can occur without detectable alterations of the consensus genomic sequence. The investigation of other possible viral factors associated with the initial steps of fitness recovery, showed that viral quasispecies heterogeneity increased between the initial clones and the passaged populations. A direct statistical correlation between viral heterogeneity and viral fitness was obtained. CONCLUSIONS: Thus, the initial fitness recovery of debilitated HIV-1 clones was mediated by an increase in quasispecies heterogeneity. This observation, together with the invariance of the consensus sequence despite fitness increases demonstrates the relevance of quasispecies heterogeneity in the evolution of HIV-1 in cell culture., Work in the Centro Nacional de Microbiología was supported by grants, SAF 2005/03833, SAF 2007/61036 by the Plan Nacional del SIDA and in part by the Red Temática Cooperativa de investigación en SIDA (Red de grupos 173) of the Fondo de Investigaciones Sanitarias de la Seguridad Social (FISss) and Fundación para la Investigación y la Prevención del SIDA en España (FIPSE) grant 36558/06, 36641/07, 36779/08, 36641/07 and 360766/09. Work in the Centro de Biología Molecular “Severo Ochoa” was supported by grant BFU 2008-02816/BMC from Ministerio de Ciencia e Innovación, FIPSE 36558/06 and 360766/09 and Fundación Ramón Areces. Centro de Investigaciones Biomédica en Red (CIBER): enfermedades hepáticas y digestivas (ehd) is funded by Instituto de Salud Carlos III. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., Sí

Descriptive epidemiology of undetected depression in institutionalized older people

  • Damian, Javier
  • Pastor-Barriuso, Roberto
  • Valderrama-Gama, Emiliana
OBJECTIVE: To measure the prevalence and associated factors of undetected depression in institutionalized older people. DESIGN AND SETTING: Epidemiologic cross-sectional study in nursing homes and residential facilities. PARTICIPANTS: A stratified cluster sample of residents 65 years of age and older living in institutions of Madrid (Spain). MEASUREMENTS: Residents were considered to be depressed if they met at least 1 of the following 3 criteria: 10-item Geriatric Depression Scale score of 4 or higher, physician's diagnosis, or antidepressant use. Prevalence of undetected depression was defined as the proportion of depressed residents without documented diagnosis or treatment. RESULTS: A total of 255 of 579 residents had depression (weighted prevalence 46.1%, 95% confidence interval [CI] 41.0%-51.3%) and 108 depressed residents were undetected (undetection prevalence 41.5%, 95% CI 33.2%-50.2%). Undetection was lower in younger residents, private versus public facilities (sex-, age-, and size-adjusted prevalence ratio [PR] 0.59, 95% CI 0.37-0.94), and larger facilities (sex-, age-, and ownership-adjusted PR 0.94 per 50-bed increase, 95% CI 0.88-1.00). Undetected depression was higher in residents with poor self-rated health (sex- and age-adjusted PR 1.83, 95% CI 1.24-2.73), whereas the opposite came about for physician-rated health (PR 0.65, 95% CI 0.44-0.95). Undetection decreased 11% (95% CI 4%-17%) per 1-medication increase, and it was lower in patients with Alzheimer disease, anxiety, and arrhythmia. CONCLUSIONS: Number of medications and self-rated health were the main determinants of undetected depression. Physician-rated health, facility characteristics (size and ownership), and some diseases could also be considered., This work was supported by the Spanish "Fondo de Investigación Sanitaria" (grant 96/0201), Sí

Proteomic strategies in the search of new biomarkers in atherothrombosis

  • Tuñón, José
  • Martín-Ventura, José Luis
  • Blanco-Colio, Luis Miguel
  • Lorenzo, Oscar
  • Lopez, Juan Antonio
  • Egido, Jesús
Extensive research has focused on the identification of novel plasma biomarkers to improve our ability to predict cardiovascular events in atherothrombosis. However, classical techniques can only assess a limited number of proteins at a time. Given that plasma contains more than 900,000 proteins, this approach will be extremely time-consuming. Novel proteomic approaches make it possible to compare the expression of hundreds of proteins in several samples in a single experiment. The classical approach consists of separation of proteins on a 2-dimensional gel followed by protein identification with mass spectrometry, although new complementary gel-free techniques are emerging. We can thus compare protein expression in an atherosclerotic plaque with that in a normal artery or study plasma proteins in patients with atherothrombosis as compared with healthy subjects. For such approaches, it is not necessary to study the published data to select potential biomarkers. However, because the number of patients that can be studied with most of these techniques is limited, what is really important is the design of the studies, assessing carefully what kind of patients should be included to obtain valid conclusions. Clinicians should thus play a key role in this design along with the basic scientist. In this article, we review several proteomic strategies carried out by our group and others, and we make a call for collaboration between clinicians and experts in proteomics. This collaboration could greatly increase the likelihood of identifying new prognostic biomarker panels in atherothrombosis and other cardiovascular disorders., This work was supported by SAF (2007/63648 and 2007/60896), CAM (S2006/GEN-0247), FIS (PI050451, PS09/01405 and CP04/ 00060), European Network (HEALTH F2-2008-200647), EUROSALUD (EUS2008-03565), Fundacion Ramon Areces, Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III, Red RECAVA (RD06/0014/0035), Fundacion Española del Corazon, Sociedad Española de Arteriosclerosis, Mutua Madrileña Automovilista and Pfizer. The CNIC is supported by the Ministerio de Ciencia e Innovacion and the Fundacion ProCNIC., Sí

Posibilidades de mejora en los planes de prevención frente al exceso de temperaturas

  • Montero Rubio, Juan Carlos
  • Mirón Pérez, Isidro J
  • Criado-Alvarez, Juan José
  • Linares-Gil, Cristina
  • Díaz Jiménez, Julio
En el verano de 2003 las temperaturas alcanzadas fueron responsables de un gran número de muertes en el continente europeo. Un año después muchos países habían implantado algún tipo de plan de prevención frente al exceso de temperaturas. Planes que ya habían mostrado su capacidad de prevenir una buena parte de la mortalidad evitable en otras latitudes. Desde entonces, son numerosos los estudios publicados ofreciendo nuevos datos sobre los efectos en salud pública de una ola de calor, que pueden contribuir a aumentar la eficiencia de estos planes de prevención. Conocer las situaciones meteorológicas de mayor riesgo, definir ola de calor o tener en cuenta el tiempo que el plan debe estar activo a partir del estudio de la relación entre la temperatura y sus efectos en salud, identificar las características meteorológicas que modulan la relación entre la temperatura y la mortalidad, localizar el perfil de las personas de mayor riesgo o diseñar protocolos de actuación con la mayor precisión posible y basado en el conocimiento científico son elementos extraídos de numerosos trabajos realizados en los últimos años que deben ser tenidos en cuenta., Sí

Pitfalls in spontaneous in vitro transformation of human mesenchymal stem cells

  • Garcia, S
  • Bernad, A
  • Martín, M C
  • Cigudosa, J C
  • Garcia-Castro, Javier
  • de la Fuente, R
Spontaneous in vitro transformation of human primary cells was, and continues to be, a scarcely described phenomenon. Only the description of the generation of the HaCAT cell line [1] is a canonical example, worldwide accepted. More recent examples included the emergence of tumorogenic populations upon in vitro culture of fetal human mesenchymal stem cells (hMSC), induced by GM-CSF and IL-4, [2] and bone marrow hMSC [3]. Other examples have also been reported after very long-term in vitro culture of telomerized hMSCTerT [4]. In this scenario, our previous results [5-7] were only a new observation to be added to this list. Only very recently similar results have been published [8], reporting spontaneous malignant transformation in 46 % of bone marrow–derived hMSC long-term cultures (5–106 weeks). However, other authors reported opposite results., Sí

Data integration and exploration for the identification of molecular mechanisms in tumor-immune cells interaction

  • Mlecnik, Bernhard
  • Sanchez-Cabo, Fatima
  • Charoentong, Pornpimol
  • Bindea, Gabriela
  • Pagès, Franck
  • Berger, Anne
  • Galon, Jerome
  • Trajanoski, Zlatko
Cancer progression is a complex process involving host-tumor interactions by multiple molecular and cellular factors of the tumor microenvironment. Tumor cells that challenge immune activity may be vulnerable to immune destruction. To address this question we have directed major efforts towards data integration and developed and installed a database for cancer immunology with more than 1700 patients and associated clinical data and biomolecular data. Mining of the database revealed novel insights into the molecular mechanisms of tumor-immune cell interaction. In this paper we present the computational tools used to analyze integrated clinical and biomolecular data. Specifically, we describe a database for heterogeneous data types, the interfacing bioinformatics and statistical tools including clustering methods, survival analysis, as well as visualization methods. Additionally, we discuss generic issues relevant to the integration of clinical and biomolecular data, as well as recent developments in integrative data analyses including biomolecular network reconstruction and mathematical modeling., This work was supported by the Austrian Ministry for Science and Research, GEN-AU Project Bioinformatics Integration Network (BIN), Austrian Science Fund (SFB Project Lipotoxicity), INSERM, the National Cancer Institute (INCa), Association pour la Recherche sur le Cancer (ARC), the Cancéropole Ile de France, Ville de Paris, and by the European Commission (FP7, Geninca Consortium, grant number 202230)., Sí

Proposal for a revised taxonomy of the family Filoviridae: classification, names of taxa and viruses, and virus abbreviations

  • Kuhn, Jens H
  • Becker, Stephan
  • Ebihara, Hideki
  • Geisbert, Thomas W
  • Johnson, Karl M
  • Kawaoka, Yoshihiro
  • Lipkin, W Ian
  • Negredo-Anton, Ana Isabel
  • Netesov, Sergey V
  • Nichol, Stuart T
  • Palacios, Gustavo
  • Peters, Clarence J
  • Tenorio, Antonio
  • Volchkov, Viktor E
  • Jahrling, Peter B
The taxonomy of the family Filoviridae (marburgviruses and ebolaviruses) has changed several times since the discovery of its members, resulting in a plethora of species and virus names and abbreviations. The current taxonomy has only been partially accepted by most laboratory virologists. Confusion likely arose for several reasons: species names that consist of several words or which (should) contain diacritical marks, the current orthographic identity of species and virus names, and the similar pronunciation of several virus abbreviations in the absence of guidance for the correct use of vernacular names. To rectify this problem, we suggest (1) to retain the current species names Reston ebolavirus, Sudan ebolavirus, and Zaire ebolavirus, but to replace the name Cote d'Ivoire ebolavirus [sic] with Taï Forest ebolavirus and Lake Victoria marburgvirus with Marburg marburgvirus; (2) to revert the virus names of the type marburgviruses and ebolaviruses to those used for decades in the field (Marburg virus instead of Lake Victoria marburgvirus and Ebola virus instead of Zaire ebolavirus); (3) to introduce names for the remaining viruses reminiscent of jargon used by laboratory virologists but nevertheless different from species names (Reston virus, Sudan virus, Taï Forest virus), and (4) to introduce distinct abbreviations for the individual viruses (RESTV for Reston virus, SUDV for Sudan virus, and TAFV for Taï Forest virus), while retaining that for Marburg virus (MARV) and reintroducing that used over decades for Ebola virus (EBOV). Paying tribute to developments in the field, we propose (a) to create a new ebolavirus species (Bundibugyo ebolavirus) for one member virus (Bundibugyo virus, BDBV); (b) to assign a second virus to the species Marburg marburgvirus (Ravn virus, RAVV) for better reflection of now available high-resolution phylogeny; and (c) to create a new tentative genus (Cuevavirus) with one tentative species (Lloviu cuevavirus) for the recently discovered Lloviu virus (LLOV). Furthermore, we explain the etymological derivation of individual names, their pronunciation, and their correct use, and we elaborate on demarcation criteria for each taxon and virus., Sí

Immunotherapy with myeloid cells for tolerance induction

  • Rodriguez-Garcia, Mercedes
  • Boros, Peter
  • Bromberg, Jonathan S
  • Cano-Ochando, Jordi
PURPOSE OF REVIEW: Understanding the interplay between myeloid dendritic cells and T cells under tolerogenic conditions, and whether their interactions induce the development of antigen-specific regulatory T cells (Tregs) is critical to uncover the mechanisms involved in the induction of indefinite allograft survival. RECENT FINDINGS: Myeloid dendritic cell-T-cell interactions are seminal events that determine the outcome of the immune response, and multiple in-vitro protocols suggest the generation of tolerogenic myeloid dendritic cells that modulate T-cell responses, and determine the outcome of the immune response to an allograft following adoptive transfer. We believe that identifying specific conditions that lead to the generation of tolerogenic myeloid dendritic cells and Tregs are critical for the manipulation of the immune response towards the development of transplantation tolerance. SUMMARY: We summarize recent findings regarding specific culture conditions that generate tolerogenic myeloid dendritic cells that induce T-cell hyporesponsiveness and Treg development, which represents a novel immunotherapeutic approach to promote the induction of indefinite graft survival prolongation. The interpretations presented here illustrate that different mechanisms govern the generation of tolerogenic myeloid dendritic cells, and we discuss the concomitant therapeutic implications., This work was supported by the Programa Ramón y Cajal RYC-2006-1588, Ministerio de Educa-ción y Ciencia SAF2007-63579, Programa José Castillejo JC2008-00065, and Programa de Investigación de Grupos Emergentes del ISCIII (to J.C.O.), and NIH R01 AI-41428, AI-72039, and the Emerald Foundation (to J.S.B.)., Sí

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