Infecções respiratórias de origem viral mais frequentes na comunidade

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Non-Viral Gene Therapy to Cells of the bovine Intervertebral Disc

Background. Low back pain is an increasing global health problem, which is associated with intervertebral disc (IVD) damage and degeneration. Major changes occur in the nucleus pulposus (NP), with the degradation of the extracellular matrix (ECM).1 Further studies showed that growth factors from transforming growth factor β (TGFβ) and bone morphogenic proteins (BMP) family may induce chondrogenic differentiation of mesenchymal stem cells (MSC).2 Focusing on non-viral gene therapies and their possible translation into the clinics, we investigated if GDF6 (syn. BMP13 or CDMP2) can induce regeneration of degraded NP. We hypothesized that IVD transfected with plasmid over-expressing GDF6 also up-regulates other NP- and chondrogenic cell markers and enhances ECM deposition. Methods. Bovine nucleus pulposus (bNPC) and annulus fibrosus cells (bAFC) were harvested from bovine coccygeal IVD. Primary cells were then electroporized with plasmid GDF6 (Origene, vector RG211366) by optimizing parameters using the Neon Transfection system (Life Technologies, Basel). After transfection, cells were cultured in 2D monolayer or 3D alginate beads for 7, 14 or 21 days. Transfection efficiency of pGDF6 was analyzed by immunohistochemistry and fluorescent microscopy. Cell phenotype was quantified by real-time RT-PCR. To test a non-viral gene therapy applied directly to 3D whole organ culture, coccygeal bovine IVDs were harvested as previously described. Bovine IVDs were transfected by injection of plasmid GDF6 into the center. Electroporation was performed with ECM830 Square Wave Electroporation System (Harvard Apparatus, MA) using 2-needle array electrode or tweezertrodes. 72 h after tranfection discs were fixed and cryosectioned and analyzed by immunofluorescence against GDF6. Results. RT-PCR and immunohistochemistry confirmed up-regulation of GFP and GDF6 in the primary bNPC/bAFC culture. The GFP-tagged GDF6 protein, however, was not visible, possibly due to failure of dimer formation as a result of fusion structure. Organ IVD culture transfection revealed GDF6 positive staining in the center of the disc using 2-needle array electrode. Results from tweezertrodes did not show any GDF6 positive cells. Conclusion. Non-viral transfection is an appealing approach for gene therapy as it fulfills the translational safety aspects of transiency and lacks the toxic effects of viral transduction. We identified novel parameters to successfully transfect primary bovine IVD cells. For transfection of whole IVD explants electroporation parameters need to be further optimized. Acknowledgements. This project was funded by the Lindenhof Foundation (Funds “Research & Teaching”) Project no. 13-02-F. The imaging part of this study was performed with the facility of the Microscopy Imaging Center (MIC), University of Bern. References. Roughly PJ (2004): Spine (Phila), 29:2691-2699 Clarke LE, McConell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014), Arthritis Research & Therapy, 16:R67

HIV infection, viral hepatitis and liver fibrosis among prison inmates in West Africa.

BACKGROUND Prisoners represent a vulnerable population for blood-borne and sexually transmitted infections which can potentially lead to liver fibrosis and ultimately cirrhosis. However, little is known about the prevalence of liver fibrosis and associated risk factors among inmates in sub-Saharan Africa. METHODS Screening of liver fibrosis was undertaken in a randomly selected sample of male inmates incarcerated in Lome, Togo and in Dakar, Senegal using transient elastography. A liver stiffness measurement ≥9.5 KPa was retained to define the presence of a severe liver fibrosis. All included inmates were also screened for HIV, Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) infection. Substances abuse including alcohol, tobacco and cannabis use were assessed during face-to-face interviews. Odds Ratio (OR) estimates were computed with their 95 % Confidence Interval (CI) to identify factors associated with severe liver fibrosis. RESULTS Overall, 680 inmates were included with a median age of 30 years [interquartile range: 24-35]. The prevalence of severe fibrosis was 3.1 % (4.9 % in Lome and 1.2 % in Dakar). Infections with HIV, HBV and HCV were identified in 2.6 %, 12.5 % and 0.5 % of inmates, respectively. Factors associated with a severe liver fibrosis were HIV infection (OR = 7.6; CI 1.8-32.1), HBV infection (OR = 4.8; CI 1.8-12.8), HCV infection (OR = 52.6; CI 4.1-673.8), use of traditional medicines (OR = 3.7; CI 1.4-10.1) and being incarcerated in Lome (OR = 3.3; CI 1.1-9.8) compared to Dakar. CONCLUSIONS HIV infection and viral hepatitis infections were identified as important and independent determinants of severe liver fibrosis. While access to active antiviral therapies against HIV and viral hepatitis expands in Africa, adapted strategies for the monitoring of liver disease need to be explored, especially in vulnerable populations such as inmates.

Memoria sobre el Establecimiento de Baños del R. sitio de Solán de Cabras : 1863 /

Incluye cuadro estadistico.

Memoria anual de las aguas y baños minero medicinales del real sitio de Solan de Cabras correspondiente á la temporada balnearia de 1866 /

Encuadernado en cartón jaspeado.

Memoria sobre la fiebre amarilla observada en España desde la entrada del presente siglo hasta el dia de hoy : en que se considera esta enfermedad como exótica, esencialmente contagiosa y trasportable por sus semilla á largas distancias.

Mode of access: Internet.

¿Es siempre el cancer una enfermedad constitucional? [Manuscrito] /

Mode of access: Internet.

Respuesta al papel que ha dado al público ... Antonio María Herrero ... sobre la enfermedad que quitó la vida a Manuel Rodriguez /

Mode of access: Internet.

Historia de la disputa, que sobre la enfermedad que quitó la vida a Manuel Rodriguez en el Hospital General de esta Corte, tuvieron el doctor Don Bernardo Lopez de Araujo, y Don Antonio Maria Herrero.

Mode of access: Internet.

Informe al publico Discurso Medico, apologético, y crítico que sobre la enfermedad, que padeció Carlos Ferrer ...

Mode of access: Internet.

Informe medico-moral de la penosissima, y rigorosa enfermedad de la epilepsia /

Mode of access: Internet.

Historia de una enfermedad freqüente, aunque poco conocida, propia del sistema linfático y consideraciones generales sobre las afecciones de los absorventes /

Mode of access: Internet.

Enlace que tiene la vida con la respiracion, ó Experimentos acerca de los efectos que producen en los animales vivos la sumersion, la estrangulacion, y las diversas especies de gases mefíticos, con la definición exacta del género de enfermedad que resulta en semejantes casos, síntomas que la distinguen de la muerte, y medios mas adequados de evitarla /

Mode of access: Internet.

Relacion historica del extraordinario phenomeno medico, que ha ocurrido en el Hospital General con un enfermo, que entró en él à 22. de enero de este año, y dictamen que sobre el conocimiento pronostico, y curacion de su enfermedad han dado los medicos de numero de los Reales Hospitales General, y Passion de esta Corte.

Mode of access: Internet.