distribution and bioactivity of the ret specific d4 aptamer in three dimensional collagen gel cultures

The success of tyrosine kinase inhibitors in cancer therapy prompted intensive research efforts addressed to the development of new specific diagnostics and therapeutics. Targeting large transmembrane molecules, including receptor tyrosine kinases, is a major pharmacologic challenge. The D4 RNA-aptamer, isolated applying the Systematic Evolution of Ligand by Exponential Enrichment procedure on living cells, has been proven a specific inhibitor of the human receptor tyrosine kinase Ret. In our attempts to generate new powerful probes for in vivo applications, in the present study, we addressed the ability of D4 to preserve its biological activity in cells embedded in three-dimensional collagen gels. These matrices provide a microenvironment mimicking the cell organization as seen in vivo , thus representing a suitable tool to approach the use of the aptamer in vivo . By taking advantage of transformed fibroblasts expressing Ret as a model system, we showed that the cells maintain normal phenotype and growth patterns when cultured in three-dimensional matrices and that the D4 aptamer preserves its ability to inhibit Ret on the surface of the cells embedded in collagen. Because the biological activity of RNA aptamers is largely dictated by their folded structure, the results indicate that a folded conformation of D4 responsible of its inhibiting function is preserved in the three-dimensional constructs, thus supporting its use in tumors in vivo . [Mol Cancer Ther 2008;7(10):3381–8

In vivo imaging of pyrrole-imidazole polyamides with positron emission tomography

The biodistribution profiles in mice of two pyrrole-imidazole polyamides were determined by PET. Pyrrole-imidazole polyamides are a class of small molecules that can be programmed to bind a broad repertoire of DNA sequences, disrupt transcription factor-DNA interfaces, and modulate gene expression pathways in cell culture experiments. The 18F-radiolabeled polyamides were prepared by oxime ligation between 4-[18F]-fluorobenzaldehyde and a hydroxylamine moiety at the polyamide C terminus. Small animal PET imaging of radiolabeled polyamides administered to mice revealed distinct differences in the biodistribution of a 5-ring β-linked polyamide versus an 8-ring hairpin, which exhibited better overall bioavailability. In vivo imaging of pyrrole-imidazole polyamides by PET is a minimum first step toward the translation of polyamide-based gene regulation from cell culture to small animal studies

Differences in risk behaviours and HIV status between primary amphetamines and opioid injectors in Estonia and Russia

Background and objective People who inject drugs (PWID) account for over half of new HIV infections in Eastern Europe and central Asia, where opioids continue to be the dominant illicit drugs injected. Stimulants including amphetamines (ATS) have been associated with HIV infection risk in several settings. We sought to examine whether primary ATS injection was associated with greater HIV risk, compared to opioid injection in two European locales with significant HIV epidemics. Methods PWID in Kohtla-Järve and St. Petersburg were recruited using respondent-driven sampling in 2012–2013. Survey data on demographic characteristics, service use, injecting and sexual risk behaviours and HIV-status (and HCV in Kohtla-Järve) were compared between primary opioid and ATS injectors using logistic regression models. Results Of 591 injectors recruited in Kohtla-Järve and 811 in St. Petersburg, 195 (33%) and 27 (4%) primarily injected ATS in each city. In both cities, ATS injectors were younger than opioid injectors, initiated injection later, injected less frequently and were more likely to have been paid for sex. In both cities, PWID had high levels of multiple sex partners. In Kohtla-Järve, ATS-injectors had lower odds of back-loading and greater odds of polydrug use than opioid-injectors. In St. Petersburg, where over half of PWID reported unsafe sharing practices, ATS-injectors were less likely to report these practices. ATS-injection was negatively associated with being HIV positive in Kohtla-Järve (aOR = 0.6; 95%CI: 0.5–0.8) and St. Petersburg (aOR = 0.3; 95%CI: 0.1–0.7). ATS-injection was negatively associated with HCV-reactivity in Kohtla-Järve (aOR = 0.5; 95%CI: 0.3–0.6). Conclusions In both locations, primary ATS injection was associated with lower injecting risk behaviours, lower odds of HIV and being paid for sex compared to opioid injection. Interventions targeting the characteristics and needs of ATS injectors are needed to increase contact with services and reduce sexual and injecting risk. Harm reduction services, including sexual risk reduction, need to be expanded for all PWID in St. Petersburg

Provincia de Tierra del Fuego, Antártida e Islas del Atlántico Sur, República Argentina

Fil: Anselmi, Gabriela. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geología Regional; Argentina.Fil: Negro, Claudia. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geología Regional; Argentina.Fil: Hernández, Mariano. Servicio Geológico Minero Argentino. Centro Regional General Roca; Argentina.Fil: Ramé, Gustavo A. Servicio Geológico Minero Argentino. Centro Regional Córdoba; Argentina.Fil: Cegarra, Marcelo I. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geología Regional; Argentina.Fil: Cristallini, Ernesto O. Instituto de Estudios Andinos "Don Pablo Groeber" IDEAN (UBA-CONICET).Fil: Benítez, Javier. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geomática; Argentina.Fil: Pucheta, Alicia. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geomática; Argentina.Fil: Tavitian Serrano, Ana Felisa. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geomática; Argentina.Fil: Álvarez, María Dolores. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geomática; Argentina.Fil: Dal Molin, Carlos. Servicio Geológico Minero Argentino. Instituto de Geología y Recursos Minerales, Dirección de Geología Regional; Argentina

Diseño del Módulo de Salidas Gráficas: Especificaciones Generales

Fil: Marquínez García, Jorge. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: García Manteca, Pilar. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Sánchez, D. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Candaosa, Norberto Gabriel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Silvia Beatríz. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Ferpozzi, Federico Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Olmos, María Isabel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Benítez, Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Rodríguez, Valentín. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Gabande, Liliana. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Tavitian Serrano, Ana Felisa. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Oyola, Matías. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Colina, A. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España

Protocolo para la Incorporación de las Cartas Geológicas de la República de Argentina

Fil: Marquínez García, Jorge. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: García Manteca, Pilar. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Sánchez , D. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Colina, A. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Candaosa, Norberto Gabriel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Silvia Beatríz. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Ferpozzi, Federico Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Olmos, María Isabel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Benítez, Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Rodríguez, Valentín. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Gabande, Liliana. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Tavitian Serrano, Ana Felisa. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Oyola, Matías. SEGEMAR: Servicio Geológico Minero Argentino; Argentina

Interaction of Rio1 Kinase with Toyocamycin Reveals a Conformational Switch That Controls Oligomeric State and Catalytic Activity

Rio1 kinase is an essential ribosome-processing factor required for proper maturation of 40 S ribosomal subunit. Although its structure is known, several questions regarding its functional remain to be addressed. We report that both Archaeoglobus fulgidus and human Rio1 bind more tightly to an adenosine analog, toyocamycin, than to ATP. Toyocamycin has antibiotic, antiviral and cytotoxic properties, and is known to inhibit ribosome biogenesis, specifically the maturation of 40 S. We determined the X-ray crystal structure of toyocamycin bound to Rio1 at 2.0 Å and demonstrated that toyocamycin binds in the ATP binding pocket of the protein. Despite this, measured steady state kinetics were inconsistent with strict competitive inhibition by toyocamycin. In analyzing this interaction, we discovered that Rio1 is capable of accessing multiple distinct oligomeric states and that toyocamycin may inhibit Rio1 by stabilizing a less catalytically active oligomer. We also present evidence of substrate inhibition by high concentrations of ATP for both archaeal and human Rio1. Oligomeric state studies show both proteins access a higher order oligomeric state in the presence of ATP. The study revealed that autophosphorylation by Rio1 reduces oligomer formation and promotes monomerization, resulting in the most active species. Taken together, these results suggest the activity of Rio1 may be modulated by regulating its oligomerization properties in a conserved mechanism, identifies the first ribosome processing target of toyocamycin and presents the first small molecule inhibitor of Rio1 kinase activity

Diseño del Módulo de Salidas Gráficas de la Carta Geológica

Fil: Marquínez García, Jorge. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: García Manteca, Pilar. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Sánchez, D. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Colina, A. INDUROT: Instituto de Recursos Naturales y Ordenación del Territorio – Universidad de Oviedo; España.Fil: Candaosa, Norberto Gabriel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Chavez, Silvia Beatríz. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Ferpozzi, Federico Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Olmos, María Isabel. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Benítez, Javier. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Rodríguez, Valentín. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Gabande, Liliana. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Tavitian Serrano, Ana Felisa. SEGEMAR: Servicio Geológico Minero Argentino; Argentina.Fil: Oyola, Matías. SEGEMAR: Servicio Geológico Minero Argentino; Argentina

Non-Invasive In Vivo Imaging of Calcium Signaling in Mice

Rapid and transient elevations of Ca2+ within cellular microdomains play a critical role in the regulation of many signal transduction pathways. Described here is a genetic approach for non-invasive detection of localized Ca2+ concentration ([Ca2+]) rises in live animals using bioluminescence imaging (BLI). Transgenic mice conditionally expressing the Ca2+-sensitive bioluminescent reporter GFP-aequorin targeted to the mitochondrial matrix were studied in several experimental paradigms. Rapid [Ca2+] rises inside the mitochondrial matrix could be readily detected during single-twitch muscle contractions. Whole body patterns of [Ca2+] were monitored in freely moving mice and during epileptic seizures. Furthermore, variations in mitochondrial [Ca2+] correlated to behavioral components of the sleep/wake cycle were observed during prolonged whole body recordings of newborn mice. This non-invasive imaging technique opens new avenues for the analysis of Ca2+ signaling whenever whole body information in freely moving animals is desired, in particular during behavioral and developmental studies