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 Référence des Leishmanioses, Université 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 “ERDF A Way of Making Europe” and by Grant FP7-HEALTH-2013-INNOVATION “New Medicine for Trypanosomatidic Infections” (Grant 603240). The authors acknowledge the “Fondazione Cassa di Risparmio di Modena” 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 “OneHealthdrugs” CA21111 for inspiring the research development

Reparación de defectos segmentales óseos 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 reparación de defectos segmentales óseos de tamaño crítico mediante la combinación 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 creó un defecto segmental crítico en las diáfisis de los radios. El defecto se rellenó con partículas de vidrio bioactivo. Los cabos del radio y los gránulos se envolvieron con una membrana de PTFe. Inmediatamente de finalizadas las intervenciones, cada 30 días y durante 150 días se efectuaron controles radiológicos de los miembros intervenidos. Transcurridos los 150 días de tratamiento en la totalidad de los ensayos se observó que los defectos se hallaban reparados. Las radiografias de control tomadas luego de finalizadas las operaciones mostraban a las partículas de vidrio ocupando el defecto a modo de un fino granulado; a los 30 días los gránulos no se distinguían entre sí y conformaban una masa uniforme y radiodensa que con el transcurso de los controles adquirió mayor radiodensidad; a los 150 días la imagen era la reparación de los defectos. A los 150 días los animales fueron sacrificados para efectuar análisis macro y microscópico del hueso neoformado. Los estudios histológicos efectuados en cortes sagitales que incluyeron los extremos del radio y el hueso neoformado que el hueso regenerado estaba constituido por numerosas trabéculas que contenían médula roja y presencia aislada de gránulos de vidrio. La membrana de PTFe protegió los gránulos de vidrio bioactivo mezclado con el coágulo de sangre proveniente de los cabos óseos seccionados. Las reacciones químicas de los gránulos con los líquidos tisulares se pusieron en evidencia a los 30 días cuando éstos se fusionaron, momento a partir del cual comenzó los procesos de osteoconducción y osteoinducción. Los mecanismos de proliferación y diferenciación mesenquimática que promueven y desencadenan la combinación del PTFe y los gránulos de vidrio bioactivo permitieron la reparación del defecto con la generación de hueso nuevo conteniendo médula roja. La combinación de los biomateriales promueven la inducción de tejido óseo en huesos largo

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