10 research outputs found
Proteomic contribution to the omic path for the identification of novel drugs overcoming resistance in Leishmaniasis.
LC-MS/MS proteomics for the rapid and selective screening of drug resistances in Leishmania infantum clinical isolates
Label-Free Mass Spectrometry Proteomics Reveals Different Pathways Modulated in THP-1 Cells Infected with Therapeutic Failure and Drug Resistance Leishmania infantum Clinical Isolates
As the world is facing increasing difficulties to treat leishmaniasis with current therapies, deeper investigation into the molecular mechanisms responsible for both drug resistance and treatment failure (TF) is essential in drug discovery and development. So far, few available drugs cause severe side effects and have developed several resistance mechanisms. Drug resistance and TF parasite strains from clinical isolates may have acquired altered expression of proteins that characterize specific mechanisms leading to therapy inefficacy. This work aims to identify the biochemical pathways of THP-1 human monocytes infected by different Leishmania infantum clinical isolates from patients with either resistance or with TF outcome, using whole cell differential Mass Spectrometry proteomics. We have adopted network enrichment analysis to integrate the transcriptomics and the proteomic results of infected cells studies. Transferrin receptor C (TFRC) and nucleoside diphosphate kinase 3 (NDK3) were discovered as overexpressed proteins in THP-1 cells infected with paromomycin, antimony, and miltefosine resistant L. infantum lines. The overall achievements represent founding concepts to confirm new targets involved in the parasitic drug resistance and TF mechanisms, and to consider in perspective the importance of a dual host-guest pharmacological approach to treat the acute stage of the disease.We thank Dr. F. Javier Moreno from the WHO Collaborating Center for Leishmaniasis, Instituto de Salud Carlos III (ISCIII), for providing L. infantum lines LLM2070, LLM2165, LLM2255, and LLM2221, isolated from HIV-positive patients with visceral leishmaniasis and TF, and the paromomycin-resistant L. infantum line LEM2126 (L2126) used in this study. Also, we thank Dra. Laurence Lachaud from the Centre National de RĂ©fĂ©rence des Leishmanioses, UniversitĂ© Montpellier (Montpellier, France), for providing the drug-resistant L. infantum lines used in this work: LEM3323 (L3323) and LEM5159 (L5159), which are SbIII- and Mil-resistant lines, respectively. The authors thank Dr. Stefania Ferrari for compiling the Leishmania database for protein search. L. infantum db was obtained from SwissProt (L. infantum entry, exported in FASTA format, updated January 2016, updates are ongoing). This work was supported in part by Grant RTI2018-097210-B-100 (to F.G.), funded by MCIN/AEI/10.13039/501100011033 and by âERDF A Way of Making Europeâ and by Grant FP7-HEALTH-2013-INNOVATION âNew Medicine for Trypanosomatidic Infectionsâ (Grant 603240). The authors acknowledge the âFondazione Cassa di Risparmio di Modenaâ for funding the UHPLC-ESI-HRMS Q-Exactive system at the Centro Interdipartimentale Grandi Strumenti (CIGS) of the University of Modena and Reggio Emilia. The authors thank the COST Action âOneHealthdrugsâ CA21111 for inspiring the research development
A dual-Omics approach to identify the modulated proteins/genes in THP-1 cells infected with different drug resistant L. infantum clinical isolates.
ReparaciĂłn de defectos segmentales Ăłseos con vidrio bioactivo y una membrana no absorbible
The authors inform the repair of segmental bone defects of critical size with the use of a combination of bioactive glass and a non-absorbable membrane of expanded politetrafluoroethilen (ePTF). Twenty four New Zealand rabbits - sexually mature males and females - were used. They were kept in individual cages with controlled temperature and fed ad libitum with a commercial formula. A critical segmental defect in the diaphysis of the radius was created in each rabbit. The defect was filled with bioactive glass granules with a particle size of 300 to 600 um . The extremes of the radius and the granules of bioactive glass were wrapped up with a ePTF membrane. Immediately after the surgical procedure, and every 30 days during a period of 150 days, X-ray controls of the operated limbs were carried out. The rabbits recovered total bearing between 48 - 72 hs. after the operation. After 150 days of treatment, the defects were repaired in all the rabbits. The control X-rays taken after the operation showed that the partic1es of glass occupied the defect had a thin granular shape. Thirty days later, the granules were not differentiated and made up a uniform and radio-dense mass that became more radio-dense in the following radiographic controls. After 150 days the x-ray images showed that the defects were fully repaired. At that moment, the animals were killed to make a macro- and microscopic analysis of the newly-formed bone. The sagittal cuts of the histological tissue that inc1uded the ends of the radius and the newly-formed bone showed that the segmental bone was constituted by numerous trabeculas that contained bone marrow and isolated presence of glass granules. The ePTF membrane protected the bioactive glass granules mixed with the blood clot coming from the cut bone tips. The ionic exchanges between the ions of Ca and P of the glass granules and the same ions of the tissue were evident 30 days after the operation when they fused. Since then, the processes of osteoconduction and osteoinduction started. The scarce presence of granules contained in the newly-formed bone suggest that the granules were absorbed. The bone regeneration was possible due to the protective action of the blood clot of the ePTF membrane and to the osteoinductive action of the bioactive glass granules that unfettered the mechanism of proliferation of the mesenchymal cells and their differentiation into osteoblasts and marrow tissue. The combination ofboth bio-materials fosters the induction ofbone tissue in long bonesSe informa la reparaciĂłn de defectos segmentales Ăłseos de tamaño crĂtico mediante la combinaciĂłn de vidrio bioactivo y una membrana no absorbible de politetrafluoroetileno expandido (PTFe). Se emplearon 24 conejos neocelandeses machos y hembras sexualmente maduros a los que se les creĂł un defecto segmental crĂtico en las diĂĄfisis de los radios. El defecto se rellenĂł con partĂculas de vidrio bioactivo. Los cabos del radio y los grĂĄnulos se envolvieron con una membrana de PTFe. Inmediatamente de finalizadas las intervenciones, cada 30 dĂas y durante 150 dĂas se efectuaron controles radiolĂłgicos de los miembros intervenidos. Transcurridos los 150 dĂas de tratamiento en la totalidad de los ensayos se observĂł que los defectos se hallaban reparados. Las radiografias de control tomadas luego de finalizadas las operaciones mostraban a las partĂculas de vidrio ocupando el defecto a modo de un fino granulado; a los 30 dĂas los grĂĄnulos no se distinguĂan entre sĂ y conformaban una masa uniforme y radiodensa que con el transcurso de los controles adquiriĂł mayor radiodensidad; a los 150 dĂas la imagen era la reparaciĂłn de los defectos. A los 150 dĂas los animales fueron sacrificados para efectuar anĂĄlisis macro y microscĂłpico del hueso neoformado. Los estudios histolĂłgicos efectuados en cortes sagitales que incluyeron los extremos del radio y el hueso neoformado que el hueso regenerado estaba constituido por numerosas trabĂ©culas que contenĂan mĂ©dula roja y presencia aislada de grĂĄnulos de vidrio. La membrana de PTFe protegiĂł los grĂĄnulos de vidrio bioactivo mezclado con el coĂĄgulo de sangre proveniente de los cabos Ăłseos seccionados. Las reacciones quĂmicas de los grĂĄnulos con los lĂquidos tisulares se pusieron en evidencia a los 30 dĂas cuando Ă©stos se fusionaron, momento a partir del cual comenzĂł los procesos de osteoconducciĂłn y osteoinducciĂłn. Los mecanismos de proliferaciĂłn y diferenciaciĂłn mesenquimĂĄtica que promueven y desencadenan la combinaciĂłn del PTFe y los grĂĄnulos de vidrio bioactivo permitieron la reparaciĂłn del defecto con la generaciĂłn de hueso nuevo conteniendo mĂ©dula roja. La combinaciĂłn de los biomateriales promueven la inducciĂłn de tejido Ăłseo en huesos largo
Proteomic contribution to the omic path for the identification of novel drugs overcoming resistance in Leishmaniasis
LC-MS/MS Helps Characterizing THP-1 Cells Proteome After Infection with Drug Resistant Leishmania Strains
Insights into host-target interaction from the proteome modulation analysis through untargeted LC-MS/MS Proteomics of drug resistant L infantum-THP1 infected cells
SARS-CoV-2 vaccination modelling for safe surgery to save lives: data from an international prospective cohort study
Background: Preoperative SARS-CoV-2 vaccination could support safer elective surgery. Vaccine numbers are limited so this study aimed to inform their prioritization by modelling.
Methods: The primary outcome was the number needed to vaccinate (NNV) to prevent one COVID-19-related death in 1 year. NNVs were based on postoperative SARS-CoV-2 rates and mortality in an international cohort study (surgical patients), and community SARS-CoV-2 incidence and case fatality data (general population). NNV estimates were stratified by age (18-49, 50-69, 70 or more years) and type of surgery. Best- and worst-case scenarios were used to describe uncertainty.
Results: NNVs were more favourable in surgical patients than the general population. The most favourable NNVs were in patients aged 70 years or more needing cancer surgery (351; best case 196, worst case 816) or non-cancer surgery (733; best case 407, worst case 1664). Both exceeded the NNV in the general population (1840; best case 1196, worst case 3066). NNVs for surgical patients remained favourable at a range of SARS-CoV-2 incidence rates in sensitivity analysis modelling. Globally, prioritizing preoperative vaccination of patients needing elective surgery ahead of the general population could prevent an additional 58 687 (best case 115 007, worst case 20 177) COVID-19-related deaths in 1 year.
Conclusion: As global roll out of SARS-CoV-2 vaccination proceeds, patients needing elective surgery should be prioritized ahead of the general population