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Santiago de Compostela, Spain

Galan Bartual S.,University of Santiago de Compostela | Garcia-Doval C.,University of Santiago de Compostela | Alonso J.,Complexo Hospitalario Universitario Of Santiago | Schoehn G.,Joseph Fourier University | And 3 more authors.
Protein Expression and Purification | Year: 2010

Bacteriophage T4 recognises its host cells through its long tail fibre protein gene product (gp) 37. Gp37 is a protein containing 1026 amino acids per monomer, forming a fibrous parallel homotrimer at the distal end of the long tail fibres. The other distal half-fibre protein, gp36, is much smaller, forming a trimer of 221 amino acids per monomer. Functional and structural studies of gp37 have been hampered by the inability to produce suitable amounts of it. We produced soluble gp37 by co-expression with two bacteriophage T4-encoded chaperones in a two-vector system; co-expression with each chaperone separately did not lead to good amounts of correctly folded, trimeric protein. An expression vector for the bacteriophage T4 fibrous protein chaperone gp57 was co-transformed into bacteria with a compatible bi-cistronic expression vector containing bacteriophage T4 genes 37 and 38. A six-histidine tag is encoded amino-terminal to the gp37 gene. Recombinant trimeric gp37, containing the histidine tag and residues 12-1026 of gp37, was purified from lysed bacteria by subsequent nickel-affinity, size exclusion and strong anion exchange column chromatography. Yields of approximately 4 mg of purified protein per litre of bacterial culture were achieved. Electron microscopy confirmed the protein to form fibres around 63 nm long, presumably gp36 makes up the remaining 11 nm in the intact distal half-fibre. Purified, correctly folded, gp37 will be useful for receptor-binding studies, high-resolution structural studies and for specific binding and detection of bacteria. © 2009 Elsevier Inc. All rights reserved.

Martinez E.,University of Barcelona | D'Albuquerque P.M.,University of Barcelona | Llibre J.M.,Lluita Contra la SIDA Foundation | Gutierrez F.,University Miguel Hernandez | And 8 more authors.
AIDS | Year: 2012

Background: Switching from boosted protease inhibitors (PI/r) to raltegravir (RAL) results in a better plasma lipid profile than continuing PI/r. Whether this strategy affects plasma biomarkers associated with atherosclerosis is unknown. Methods: We assessed 48-week changes in fasting lipids and several biomarkers including serum high-sensitivity C-reactive protein (hsCRP), monocyte chemoattractant protein 1 (MCP-1), osteoprotegerin, interleukin (IL) 6, IL-10, tumor necrosis factor alpha (TNF-α), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin and P-selectin, adiponectin, insulin, and D-dimer in otherwise healthy, virologically suppressed HIV-infected patients treated with PI/r who randomly switched from PI/r to RAL or continued with PI/r in the SPIRAL trial. Biomarkers and lipids at baseline and 48-week changes between both study arms were compared. Correlations between changes in biomarkers and changes in lipids were also evaluated. Results: Of 273 patients initiating study drugs in the SPIRAL trial, 233 (119 RAL, 114 PI/r) remained on allocated therapy for 48 weeks and had sera available for the purpose of this substudy. Triglycerides (-28%, P < 0.0001), total (-14%, P < 0.0001), low-density lipoprotein (-9%, P = 0.0069), and high-density lipoprotein (-10%, P = 0.0017) cholesterol decreased in RAL relative to the PI/r group. Among biomarkers, hsCRP (-40%, P < 0.0001), MCP-1 (-20%, P = 0.0003), osteoprotegerin (-13%, P = 0.0024), IL-6 (-46%,P < 0.0001), TNF-α (-27%, P = 0.0011), insulin (-26%, P < 0.0001), and D-dimer (-8%, P = 0.0187) decreased in RAL relative to PI/r group, whereas IL-10 (+1%, P = 0.7773), ICAM-1 (-6%, P = 0.1255), VCAM-1(0%, P = 0.8671), E-selectin (-9%, P = 0.2174), P-selectin (-6%, P = 0.3865), and adiponectin (+8%, P = 0.2028) remained unchanged. Biomarkers and lipids changes at 48 weeks were weakly correlated. Conclusion: Switching from PI/r to RAL induced significant changes in several cardiovascular biomarkers that were not completely explained by lipid changes. © 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Pei Y.,University of Toronto | Wang K.,University of Toronto | Garcia-Gonzalez M.,Complexo Hospitalario Universitario Of Santiago | He N.,University of Toronto | And 3 more authors.
Kidney International | Year: 2012

Mutations of PKD1 and PKD2 account for most cases of autosomal dominant polycystic kidney disease (ADPKD). Compared with PKD2, patients with PKD1 typically have more severe renal disease. Here, we report a follow-up study of a unique multigeneration family with bilineal ADPKD (NFL10) in which a PKD1 disease haplotype and a PKD2 (L736X) mutation co-segregated with 18 and 14 affected individuals, respectively. In our updated genotype-phenotype analysis of the family, we found that PKD1-affected individuals had uniformly mild renal disease similar to the PKD2-affected individuals. By sequencing all the exons and splice junctions of PKD1, we identified two missense mutations (Y528C and R1942H) from a PKD1-affected individual. Although both variants were predicted to be damaging to the mutant protein, only Y528C co-segregated with all of the PKD1-affected individuals in NFL10. Studies in MDCK cells stably expressing wild-type and mutant forms of PKD found that cell lines expressing the Y528C variant formed cysts in culture and displayed increased rates of growth and apoptosis. Thus, Y528C functions as a hypomorphic PKD1 allele. These findings have important implications for pathogenic mechanisms and molecular diagnostics of ADPKD. © 2012 International Society of Nephrology.

Cruz-Ferro E.,Galician Tuberculosis Prevention and Control Programme | Ursua-Diaz M.I.,Galician Tuberculosis Prevention and Control Programme | Taboada-Rodriguez J.A.,Galician Tuberculosis Prevention and Control Programme | Hervada-Vidal X.,Galician Tuberculosis Prevention and Control Programme | And 2 more authors.
International Journal of Tuberculosis and Lung Disease | Year: 2014

SETTING: Galicia, Spain. OBJECTIVE: To describe changes in tuberculosis (TB) epidemiology and characteristics in Galicia, Spain, during the period 1996-2011. DESIGN: Retrospective observational descriptive study of data obtained from the Tuberculosis Information System. The Galician Tuberculosis Prevention and Control Programme, created in 1994, is based in seven tuberculosis units that actively collect data on case finding and follow-up of all cases of TB in the region. RESULTS: TB incidence fell from 72.9 cases per 100 000 population in 1996 to 24.6 in 2011 (respectively 40.5 and 14.2 in patients aged <15 years). In 2011, 49.8% (n = 343) of patients were aged between 25-54 years; 62.3% (n = 429) were male; 52.1% (n = 359) had pulmonary TB (PTB) alone, of whom 33.5% (n = 144) had cavitary lesions; 50.7% (n = 218 PTB cases) were sputum smear-positive and 80.5% (346 PTB cases) were culture-positive. The median diagnostic delay was 56 days; 4.6% (n = 32) were human immunodeficiency virus positive and 5.2% (n = 36) were immigrants. The treatment success rate was close to 90%. Contacts of 86.7% of the smear-positive index cases were evaluated. CONCLUSION: TB incidence in Galicia is progressively decreasing; however, it is still higher than that of neighbouring regions. A long diagnostic delay was observed, which may have contributed to the high incidence rate in children. © 2014 The Union.

Fernandez Y.,Hospital Universitario Central Of Asturias | Cueva J.,Complexo Hospitalario Universitario Of Santiago | Palomo A.G.,Hospital de Leon | Ramos M.,Centro Oncologico Regional | And 6 more authors.
Cancer Treatment Reviews | Year: 2010

Metastatic breast cancer is ultimately an incurable disease, although recent data have shown that its incidence is decreasing and that patients with metastatic breast cancer live longer. This improvement in survival seems to be linked with the introduction of new therapeutic agents, novel combinations of existing therapies and targeted therapies. Our increasing understanding of the molecular biology of metastatic disease has allowed the development of therapies aimed at specific molecular targets. Some of these have already been approved for the treatment of metastatic breast cancer in combination with cytotoxics, and others have shown promising results regarding disease-free survival, overall response rates and time to disease progression. Given the enormous amount of information about drug discovery in cancer, it is important to be familiar with the present state of the treatment of metastatic breast cancer. The purpose of this review is to provide an update on the development of some of the most promising novel agents and treatment strategies in metastatic breast cancer. © 2009 Elsevier Ltd. All rights reserved.

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