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Acinetobacter baumannii; Antimicrobial resistance; Efflux pump inhibitors Acinetobacter baumannii; Resistencia antimicrobiana; Inhibidores de bomba de eflujo Acinetobacter baumannii; Resistència antimicrobiana; Inhibidors de bomba d'eflux (1) Background: Acinetobacter baumannii has become the most important pathogen responsible for nosocomial infections in health systems. It expresses several resistance mechanisms, including the production of β-lactamases, changes in the cell membrane, and the expression of efflux pumps. (2) Methods: A. baumannii was detected by PCR amplification of the blaOXA-51-like gene. Antimicrobial susceptibility to fluoroquinolones and aminoglycosides was assessed using the broth microdilution technique according to 2018 CLSI guidelines. Efflux pump system activity was assessed by the addition of a phenylalanine–arginine beta-naphthylamide (PAβN) inhibitor. (3) Results: A total of nineteen A. baumannii clinical isolates were included in the study. In an overall analysis, in the presence of PAβN, amikacin susceptibility rates changed from 84.2% to 100%; regarding tobramycin, they changed from 68.4% to 84.2%; for nalidixic acid, they changed from 73.7% to 79.0%; as per ciprofloxacin, they changed from 68.4% to 73.7%; and, for levofloxacin, they stayed as 79.0% in both groups. (4) Conclusions: The addition of PAβN demonstrated a decrease in the rates of resistance to antimicrobials from the family of quinolones and aminoglycosides. Efflux pumps play an important role in the emergence of multidrug-resistant A. baumannii strains, and their inhibition may be useful as adjunctive therapy against this pathogen. This research was supported through Incentive for Research of the Universidad Peruana de Ciencias Aplicadas (Nº UPC-C007-2020), Lima, Peru. The funders had no role in the study design, data collection, and analysis, nor in the decision to publish or the preparation of the manuscript.

Klebsiella; Aspiration pneumonia; Multidrug-resistance Klebsiella; Neumonía por aspiración; Resistencia a múltiples fármacos Klebsiella; Pneumònia per aspiració; Resistència a múltiples fàrmacs VAP due to multidrug-resistant (MDR) bacteria is a frequent infection among patients in ICUs. Patient characteristics and mortality in mono- and polybacterial cases of VAP may differ. A single-centre, retrospective 3-year study was conducted in the four ICUs of a Lithuanian referral university hospital, aiming to compare both the clinical features and the 60-day ICU all-cause mortality of monobacterial and polybacterial MDR Klebsiella spp. VAP episodes. Of the 86 MDR Klebsiella spp. VAP episodes analyzed, 50 (58.1%) were polybacterial. The 60-day mortality was higher (p < 0.05) in polybacterial episodes: overall (50.0 vs. 27.8%), in the sub-group with less-severe disease (SOFA < 8) at VAP onset (45.5 vs. 15.0%), even with appropriate treatment (41.7 vs. 12.5%), and the sub-group of extended drug-resistant (XDR) Klebsiella spp. (46.4 vs. 17.6%). The ICU mortality (44.0 vs. 22.5%) was also higher in the polybacterial episodes. The monobacterial MDR Klebsiella spp. VAP was associated (p < 0.05) with prior hospitalization (61.1 vs. 40.0%), diabetes mellitus (30.6 vs. 5.8%), obesity (30.6 vs. 4.7%), prior antibiotic therapy (77.8 vs. 52.0%), prior treatment with cephalosporins (66.7 vs. 36.0%), and SOFA cardiovascular ≥ 3 (44.4 vs. 10.0%) at VAP onset. Patients with polybacterial VAP were more likely (p < 0.05) to be comatose (22.2 vs. 52.0%) and had a higher SAPS II score (median [IQR] 45.0 [35.25–51.1] vs. 50.0 [40.5–60.75]) at VAP onset. Polybacterial MDR Klebsiella spp. VAP had distinct demographic and clinical characteristics compared to monobacterial, and was associated with poorer outcomes.

Treballs Finals de Grau de Química, Facultat de Química, Universitat de Barcelona, Any: 2023, Tutor: Francesc Rabanal Anglada, Since the discovery of the antibiotics, countless of human lives have been saved. The human lifespan has been increased on average by 23 years since this founding. Unfortunately, the appearance of multidrug resistant bacteria seems to be foreshadowing the dawn of the post antibiotic era. With an urgent sanitary dilemma, the necessity of the development of new antibiotics active against multidrug resistant bacteria have been declared. The WHO (World Health Organization) have outlined the growing concern on Gram-negative bacteria, which have proven to be the most resistant to current antibiotic. In this project, an analogue of a polymyxin family member called macolacin, which is active against a certain resistance shown by Gram-negative bacteria, is being synthesized and tested through the minimum inhibitory concentration. The polymyxin family is a group of a pentacationic cyclic lipo-decapeptides with substantial activity against Gram-negative bacteria being already used as a last resort drug. Unfortunately, a severe nephrotoxicity was detected in previous studies4 and the analogue synthesized is meant to keep the activity while diminishing its toxicity through a disulfide bond. In addition, a 14 amino acids cyclic peptide developed by Polyphor called murepavadin which is extremely active against P. aeruginosa but with some nephrotoxicity will be synthesized. The peptides are being synthesized through solid phase peptide synthesis with the Fmoc/tBu orthogonal strategy being used. Both peptides being synthesized are being characterized through HPLC-MS.

Cistitis aguda no complicada; Antibiòtics; Infeccions Cistitis aguda no complicada; Antibióticos; Infecciones Uncomplicated acute cystitis; Antibiotics; Infections Aquest document informa que en dones amb cistitis aguda no complicada, no es recomana el tractament amb fluoroquinolones ni l’associació amoxicil·lina/àcid clavulànic com a primera opció. Este documento informa que en mujeres con cistitis aguda no complicada, no se recomienda el tratamiento con fluoroquinolonas ni la asociación amoxicilina/ácido clavulánico como primera opción.

[spa] Uno de los problemas de salud de más relevancia a nivel global es el de las infecciones por bacterias resistentes a los antibióticos. La Organización Mundial de la Salud (OMS) junto al Centro para el Control y la Prevención de Enfermedades (CDC) consideran a los patógenos resistentes a los antibióticos como una de las amenazas inminentes para la salud humana. El listado de microorganismos llamado ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter bau-mannii, Pseudomonas aeruginosa y Enterobacter spp), recoge las especies patógenas de más relevancia por su potencial de presentar resistencia a múl-tiples antibióticos. En ese sentido, S. aureus destaca por la adquisición de mecanismos de resistencia a casi todos los antibióticos empleados para su tratamiento, como los β-lactámicos, los glicopéptidos y las oxazolidinonas. MRSA a su vez, es uno de los patógenos resistentes más relevante, con una amplia distribución y asociado a infecciones hospitalarias, comunitarias e incluso ganaderas. También destaca su gran plasticidad genética, que le permite adaptarse a condiciones ambientales diversas, su capacidad patogénica y la frecuente combinación de resistencias a otros grupos de antibióticos. La accesibilidad durante la última década a técnicas de secuenciación del genoma completo de los microorganismos y la disminución de su coste económico, han cambiado la perspectiva de los estudios epidemiológicos y de la tipificación molecular. Esta tecnología ofrece la posibilidad de analizar genomas completos con un nivel de discriminación de unos pocos nucleótidos y establecer relaciones genéticas con una precisión inalcanzable por técnicas moleculares clásicas. Con una única técnica, se puede obtener una gran cantidad de información para el estudio de genes relacionados con la resistencia antibiótica, la producción de toxinas, la adherencia bacteriana y estudios de elementos móviles y fijos. Por ello, esta tesis doctoral pretende analizar la epidemiología de las infecciones por MRSA y las características clínicas de la infección bacteriémica; la dinámica poblacional de los clones de MRSA y su evolución en el tiempo y, por último, la base molecular de la resistencia a los antibióticos en MRSA, mediante la incorporación de tecnologías de secuenciación del genoma completo.

Breast cancer; Heterodimerization Cáncer de mama; Heterodimerización Càncer de mama; Heterodimerització Anti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, different mechanisms play a role in treatment resistance. Here, we identified AXL overexpression as an essential mechanism of trastuzumab resistance. AXL orchestrates epithelial-to-mesenchymal transition and heterodimerizes with HER2, leading to activation of PI3K/AKT and MAPK pathways in a ligand-independent manner. Genetic depletion and pharmacological inhibition of AXL restored trastuzumab response in vitro and in vivo. AXL inhibitor plus trastuzumab achieved complete regression in trastuzumab-resistant patient-derived xenograft models. Moreover, AXL expression in HER2-positive primary tumors was able to predict prognosis. Data from the PAMELA trial showed a change in AXL expression during neoadjuvant dual HER2 blockade, supporting its role in resistance. Therefore, our study highlights the importance of targeting AXL in combination with anti-HER2 drugs across HER2-amplified breast cancer patients with high AXL expression. Furthermore, it unveils the potential value of AXL as a druggable prognostic biomarker in HER2-positive breast cancer. A.A.-A., E.J.A., and F.B.-M. were supported by Asociación Española contra el Cáncer AECC (PRDVA18013LLUC to A.A.-A., POSTD211413AREN to E.J.A., and AECC_Postdoctoral17-1062 to F.B.-M.). A.M.-S. was funded by the Spanish Government (PFIS FI20/00188). J.Ar. is supported by Breast Cancer Research Foundation (BCRF-20-08), Instituto de Salud Carlos III Project reference number AC15/00062, and the EC under the framework of the ERA-NET TRANSCAN-2 initiative cofinanced by FEDER, Instituto de Salud Carlos III (CB16/12/00449 and PI19/01181), and Asociación Española Contra el Cáncer (AECC). A.P. was supported by Instituto de Salud Carlos III—PI19/01846, Breast Cancer Now—2018NOVPCC1294. P.E. and A.L. were funded by Instituto de Salud Carlos III and cofinanced by FEDER (PI18/01219 to P.E. and CB16/12/00481 to A.L.). J.M.C. was funded by Sociedad Española de Oncología Médica (Rio Hortega-SEOM) and Compromiso ADAMED.

Tetracycline resistance in streptococci is mainly due to ribosomal protection mediated by the tet(M) gene that is usually located in the integrative and conjugative elements (ICEs) of the Tn916-family. In this study, we analyzed the genes involved in tetracycline resistance and the associated mobile genetic elements (MGEs) in Streptococcus dysgalactiae subsp. equisimilis (SDSE) causing invasive disease. SDSE resistant to tetracycline collected from 2012 to 2019 in a single hospital and from 2018 in three other hospitals were analyzed by whole genome sequencing. Out of a total of 84 SDSE isolates, 24 (28.5%) were resistant to tetracycline due to the presence of tet(M) (n = 22), tet(W) (n = 1), or tet(L) plus tet(W) (n = 1). The tet(M) genes were found in the ICEs of the Tn916-family (n = 10) and in a new integrative and mobilizable element (IME; n = 12). Phylogenetic analysis showed a higher genetic diversity among the strains carrying Tn916 than those having the new IME, which were closely related, and all belonged to CC15. In conclusion, tetracycline resistance in SDSE is mostly due to the tet(M) gene associated with ICEs belonging to the Tn916-family and a new IME. This new IME is a major cause of tetracycline resistance in invasive Streptococcus dysgalactiae subsp. equisimilis in our settings.

Resistència antibiòtica; Neisseria meningitidis; Salut pública Resistencia antibiótica; Neisseria meningitidis; Salud pública Antibiotic resistance; Neisseria meningitidis; Public health Aquest informe té com a objectiu analitzar les característiques epidemiològiques dels casos confirmats de N. meningitidis i analitzar la sensibilitat antimicrobiana dels casos declarats a l’SNMC durant els anys 2016 – 2019.

Pathogen-focused, randomized, controlled trials (PF-RCT) are important in the fight against carbapenem-resistant (CR) Gram-negative infections. Some recently approved antibiotics and older generic antibiotics with activity against CR Gram-negative bacteria were investigated in PF-RCTs in a variety of infections. We searched Pubmed, Cochrane database and international clinical trial databases for PF-RCTs for the period between 2005 and 2020 and compared the study designs, patient populations, infection types, pathogens, and Day-28 all-cause mortality (ACM). PF-RCTs are particularly challenging to quantitatively assess and compare due to the heterogeneity in infection types, pathogens, CR mechanism, inclusion/exclusion criteria, and endpoints. Data interpretation is further complicated by lack of formal statistical analysis plans and/or non-inferiority design, and limited power across most PF-RCTs. The studies with new antibiotics (i.e. plazomicin, meropenem/vaborbactam, cefiderocol) ranked lower regarding feasibility, with relatively small sample sizes (analyzed: 37–118) versus the comparative effectiveness studies of older generic drugs (analyzed: 94–406). ACM ranged between 11.8% and 40% for CR Enterobacterales, 17.7% and 57.4% for CR Acinetobacter spp., and 20.0% and 30.8% for CR Pseudomonas aeruginosa. The information gathered must be considered carefully alongside the study limitations and caution should be exercised when making direct comparisons across trials. PLAIN LANGUAGE SUMMARY New antibiotics to treat multidrug-resistant Gram-negative bacterial infections are needed because antimicrobial resistance has become a global threat. In recent years, several pathogen-focused, randomized, controlled clinical trials were conducted to test new antibiotics or combinations of older generic antibiotics in the fight against resistant bacteria. However, these trials were exceptionally challenging and most of them enrolled relatively few patients. These studies were highly heterogeneous in terms of species, antibiotics, infection site, mechanism of resistance, endpoints and patient factors. In these trials, all-cause mortality at Day 28 or Day 30 were numerically lower with the new antibiotics in infections caused by carbapenem-resistant (CR) Enterobacterales. However, in the trials which investigated CR Acinetobacter spp. infections, there was no reduction in all-cause mortality at Day 28 or Day 30 with combinations of older generic antibiotics compared with colistin monotherapy. Limited information was available for CR Pseudomonas aeruginosa. More pathogen-focused, randomized, controlled clinical trials with more feasible design and higher patient numbers are needed to demonstrate clinical benefit in drug-resistant infections.

Cancer; Cell biology Càncer; Biologia cel·lular Cáncer; Biología celular Resistance to therapy remains a major obstacle in cancer management. Although treatment with hormone and CDK4/6 inhibitors is successful in luminal breast cancer, resistance to these treatments is frequent, highlighting the need for novel therapeutic strategies to delay disease progression and improve patient survival. Here, we assessed the mechanisms of acquired resistance using T47D and MCF-7 tamoxifen- and palbociclib-resistant cell-line variants in culture and as xenografts, and patient-derived cells (PDCs) obtained from sensitive or resistant patient-derived xenografts (PDXs). In these models, we analyzed the effect of specific kinase inhibitors on survival, signaling and cellular aggressiveness. Our results revealed that mTOR inhibition is more effective than PI3K inhibition in overcoming resistance, irrespective of PIK3CA mutation status, by decreasing cell proliferation and tumor growth, as well as reducing cell migration and stemness. Moreover, a combination of mTOR and CDK4/6 inhibitors may prevent pathway reactivation downstream of PI3K, interfering with the survival of resistant cells and consequent tumor escape. In conclusion, we highlight the benefits of incorporating mTOR inhibitors into the current therapy in ER + breast cancer. This alternative therapeutic strategy not only enhances the antitumor response but may also delay the emergence of resistance and tumor recurrence. This work was supported by CONICET, ANPCYT (Grants PICT2509 & PICT0345), Instituto Nacional del Cáncer (Grants 2016 & 2018), Fundación Williams, Fundación Bunge & Born (Oster Grant) (Argentina); Instituto de Salud Carlos III (Grants PI20/00892 & CPII19/0033), Ministerio de Economía y Competitividad (FJCI-2015-25412) (Spain).

Simple Summary After the recent irruption of asciminib into the therapeutic arsenal for chronic myeloid leukemia, real-life data remain scarce to determine which patients may benefit most from this drug. Data on the efficacy of the drug in real-world setting have been reported, but a detailed analysis of the toxicity profile and the influence of prior intolerance to classical tyrosine kinase inhibitors (TKIs) has not been performed. The aim of the present analysis is to study in detail the toxicity profile of asciminib as well as to describe the risk of cross-toxicity with classical TKIs. These results may help to select the patient profile with the best chance of therapeutic success with asciminib monotherapy. (1) Background: Despite the prognostic improvements achieved with tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia (CML), a minority of patients still fail TKIs. The recent introduction of asciminib may be a promising option in intolerant patients, as it is a first-in-class inhibitor with a more selective mechanism of action different from the ATP-competitive inhibition that occurs with TKIs. Therefore, our goal was to analyze toxicities shown with asciminib as well as to study cross-toxicity with previous TKIs. (2) Methods: An observational, multicenter, retrospective study was performed with data from 77 patients with CML with therapeutic failure to second-generation TKIs who received asciminib through a managed-access program (MAP) (3) Results: With a median follow-up of 13.7 months, 22 patients (28.5%) discontinued treatment: 32% (7/22) due to intolerance and 45% (10/22) due to resistance. Fifty-five percent of the patients reported adverse effects (AEs) with asciminib and eighteen percent grade 3-4. Most frequent AEs were: fatigue (18%), thrombocytopenia (17%), anemia (12%), and arthralgias (12%). None of the patients experienced cardiovascular events or occlusive arterial disease. Further, 26%, 25%, and 9% of patients required dose adjustment, temporary suspension, or definitive discontinuation of treatment, respectively. Toxicities under asciminib seemed lower than with prior TKIs for anemia, cardiovascular events, pleural/pericardial effusion, diarrhea, and edema. Cross-toxicity risk was statistically significant for thrombocytopenia, anemia, neutropenia, fatigue, vomiting, and pancreatitis. (4) Conclusion: Asciminib is a molecule with a good safety profile and with a low rate of AEs. However, despite its new mechanism of action, asciminib presents a risk of cross-toxicity with classical TKIs for some AEs.

[spa] Alrededor del 10% de los pacientes con cáncer colorrectal metastático (mCRC) presentan la mutación BRAF(V600E), mutación que está asociada a una peor respuesta al tratamiento estándar con agentes quimioterapéuticos y a una menor supervivencia. Recientemente se ha aprobado el uso de la combinación encorafenib + cetuximab para el tratamiento del mCRC BRAF(V600E). Desafortunadamente, sólo un pequeño porcentaje de los pacientes responden a este tratamiento y una importante fracción de éstos recaen al poco tiempo, lo que hace que sea muy importante identificar los mecanismos asociados a la adquisición de resistencia a los inhibidores de BRAF (BRAFi) y la identificación de nuevas vulnerabilidades. En este proyecto, hemos descrito que en respuesta a terapias combinadas con BRAFi en CRC se da una inducción del programa hipóxico y angiogénico, la activación de los fibroblastos asociados al cáncer (CAFs) y una transición a un fenotipo extremadamente mucinoso. Donde, la calidad de la ramificación de la estructura de las mucinas es necesaria para la adquisición de resistencia a las terapias BRAFi. Además, hemos observado un aumento de la expresión de MMP7, el cual promueve y mejora la liberación y accesibilidad de VEGFA bioactivo, lo que induce la angiogénesis tras el tratamiento en modelos preclínicos PDX y líneas celulares tratados con encorafenib + cetuximab + binimetinib. Estos acontecimientos generan una vulnerabilidad que nos dio la oportunidad de diseñar nuevas combinaciones racionales y efectivas añadiendo un inhibidor de VEGFA (VEGFAi) al tratamiento. Esta nueva combinación con el anticuerpo anti-angiogénico bevacizumab retrasó el tiempo de progresión de las terapias BRAFi y nos ha permitido establecer un ensayo clínico nuevo para validar este nuevo régimen terapéutico en pacientes de CRC con tumores BRAF(V600E). Paralelamente, hemos descrito que en pacientes de CRC la hipermetilación del promotor del gen BRCA1 puede reprimir su expresión, liderando así una deficiencia en la reparación por recombinación homóloga (HRD) y una sensibilidad a los inhibidores de PARP (PARPi). Además, hemos descrito de forma detallada un caso de CRC en el que el tratamiento y la progresión a los BRAFi da lugar a la selección de clones con una reparación homóloga funcional (HRP), los cuales provenían de una enfermedad que inicialmente presentaba HRD. De este caso, hemos extraído una firma de metilación que nos permitiría identificar casos con HRD de diferentes tipos tumorales. Esto es especialmente evidente en tumores donde la HRD es una alteración frecuente, como es el caso del adenocarcinoma de pulmón o el cáncer de ovario. Los datos obtenidos de este paciente apoyan el concepto de tratar a pacientes de CRC con PARPi mientras presenten una HRD. Son necesarios más estudios para refinar y validar el potencial de nuestra firma epigenética como marcador de selección de pacientes HRD sensibles a PARPi., [eng] Around 10% of patients with metastatic colorectal cancer (mCRC) present a BRAF(V600E) mutation, which is associated with a poor response to standard chemotherapy and shorter overall survival. The combination encorafenib + cetuximab has been recently approved for the treatment of mCRC BRAF(V600E). Unfortunately, only a small percentage of them respond to this treatment and a large fraction of patients relapse shortly, so it has become very important to identify the mechanisms associated to the acquired resistance to BRAF inhibitors (BRAFi) and the identification of new vulnerabilities. In this project, we described a hypoxic and angiogenic program induction, the activation of tumour associated fibroblasts (CAFs) and the transition to an extreme mucinous differentiation phenotype as a response to BRAFi- combined therapies in CRC. This drastic tumour tissue remodelling leads to a non-genetic drug resistance. In particular the quality of the branched structure in mucins is required for the acquisition of resistance to BRAFi therapies. Furthermore, we observed an increase in MMP7 expression that promoted an enhanced release of bioactive VEGFA accessible to induce angiogenesis upon treatment of preclinical PDX and cell line models with encorafenib + cetuximab + binimetinib. These events became a vulnerability that gave us the opportunity to design new rational and effective combinations adding a VEGFA inhibitor (VEGFAi) to the treatment. This new combination with the anti- angiogenic antibody bevacizumab delayed the time to progression to BRAFi therapy and led us to set a new clinical trial to validate such therapeutic regime in CRC patients with BRAF(V600E) tumours. In parallel, we describe that in CRC patients the hypermethylation of BRCA1 gene promoter can repress its expression leading to Homologous Repair Deficiency (HRD) and sensitivity to PARP inhibitors (PARPi). Here we also report in detail a CRC case in which treatment and progression to BRAF(V600E) inhibitory therapy selected Homologous Repair Proficient (HRP) clones from an initial disease presenting HRD. From this case, we extracted a methylation signature that could identify HRD cases suffering of different tumour types. This is particularly evident in tumours where HRD has been described as a frequent alteration, such as lung adenocarcinomas or ovarian cancer. Our real patient data supports the concept of treating CRC patients with PARP inhibitors as far as they present HRD. Future studies will be required to refine and validate the potential of our epigenetic signature as biomarker for selecting HRD patients sensitive to PARP inhibitors

The growth of antibiotic resistance has stimulated interest in understanding the mechanisms by which antibiotic resistance genes (ARG) are mobilized. Among them, studies analyzing the presence of ARGs in the viral fraction of environmental, food and human samples, and reporting bacteriophages as vehicles of ARG transmission, have been the focus of increasing research. However, it has been argued that in these studies the abundance of phages carrying ARGs has been overestimated due to experimental contamination with non-packaged bacterial DNA or other elements such as outer membrane vesicles (OMVs). This study aims to shed light on the extent to which phages, OMVs or contaminating non-packaged DNA contribute as carriers of ARGs in the viromes. The viral fractions of three types of food (chicken, fish, and mussels) were selected as sources of ARG-carrying phage particles, whose ability to infect and propagate in an Escherichia coli host was confirmed after isolation. The ARG-containing fraction was further purified by CsCl density gradient centrifugation and, after removal of DNA outside the capsids, ARGs inside the particles were confirmed. The purified fraction was stained with SYBR Gold, which allowed the visualization of phage capsids attached to and infecting E. coli cells. Phages with Myoviridae and Siphoviridae morphology were observed by electron microscopy. The proteins in the purified fraction belonged predominantly to phages (71.8% in fish, 52.9% in mussels, 78.7% in chicken sample 1, and 64.1% in chicken sample 2), mainly corresponding to tail, capsid, and other structural proteins, whereas membrane proteins, expected to be abundant if OMVs were present, accounted for only 3.8–21.4% of the protein content. The predominance of phage particles in the viromes supports the reliability of the protocols used in this study and in recent findings on the abundance of ARG-carrying phage particles. This work was supported by the Spanish Ministerio de Ciencia e Innovación (PID2020-113355GB-I00), the Agencia Estatal de Investigación (AEI) and the European regional fund (ERF). The study was partially supported by the Generalitat de Catalunya (2017SGR170). PB-P has a grant from the Spanish Ministry of Economy, Industry and Competitiveness (BES-2017-081296), SM-C has a grant from Colciencias (Republic of Colombia) and LR-R is lecturer of the Serra-Hunter program, Generalitat de Catalunya. MDR-B has a Margarita Salas fellowship from the Spanish Ministerio de Universidades.

Simple Summary Triple-negative breast cancer (TNBC) is currently in the clinical research spotlight because of the tumor's aggressive and invasive nature and the scarcity of therapeutic targets. Despite recent advances in identifying reliable prognostic biomarkers in the tumor microenvironment (TME), rigorous evaluation of their predictive capacity remains challenging. We describe the immune cellular and genetic profile of the residual tumor of TNBC that does not achieve a pathological complete response (pCR) after neoadjuvant chemotherapy (NAC). A high concentration of lymphocytes and dendritic cells, as well as genetic TME markers such as MUC-1 and CXCL13 in the residual tumor, are valuable prognostic factors of survival and relapse in TNBC patients. From a clinical health perspective, a thorough understanding of the composition of the TME and its prognostic implications might yield relevant immunological information and reveal key predictive biomarkers. This could ultimately help substantially improve the outcomes of residual cancer-burdened TNBC patients after NAC. With a high risk of relapse and death, and a poor or absent response to therapeutics, the triple-negative breast cancer (TNBC) subtype is particularly challenging, especially in patients who cannot achieve a pathological complete response (pCR) after neoadjuvant chemotherapy (NAC). Although the tumor microenvironment (TME) is known to influence disease progression and the effectiveness of therapeutics, its predictive and prognostic potential remains uncertain. This work aimed to define the residual TME profile after NAC of a retrospective cohort with 96 TNBC patients by immunohistochemical staining (cell markers) and chromogenic in situ hybridization (genetic markers). Kaplan-Meier curves were used to estimate the influence of the selected TME markers on five-year overall survival (OS) and relapse-free survival (RFS) probabilities. The risks of each variable being associated with relapse and death were determined through univariate and multivariate Cox analyses. We describe a unique tumor-infiltrating immune profile with high levels of lymphocytes (CD4, FOXP3) and dendritic cells (CD21, CD1a and CD83) that are valuable prognostic factors in post-NAC TNBC patients. Our study also demonstrates the value of considering not only cellular but also genetic TME markers such as MUC-1 and CXCL13 in routine clinical diagnosis to refine prognosis modelling.

Cistitis; Antibiòtics; Infeccions Cistitis; Antibióticos; Infecciones Cystitis; Antibiotics; Infections Si ets una dona adulta amb cistitis no està recomanat prendre determinats antibiòtics.

© 2021 The Authors., Bacterial infection of implanted scaffolds may have fatal consequences and, in combination with the emergence of multidrug bacterial resistance, the development of advanced antibacterial biomaterials and constructs is of great interest. Since decades ago, metals and their ions had been used to minimize bacterial infection risk and, more recently, metal-based nanomaterials, with improved antimicrobial properties, have been advocated as a novel and tunable alternative. A comprehensive review is provided on how metal ions and ion nanoparticles have the potential to decrease or eliminate unwanted bacteria. Antibacterial mechanisms such as oxidative stress induction, ion release and disruption of biomolecules are currently well accepted. However, the exact antimicrobial mechanisms of the discussed metal compounds remain poorly understood. The combination of different metal ions and surface decorations of nanoparticles will lead to synergistic effects and improved microbial killing, and allow to mitigate potential side effects to the host. Starting with a general overview of antibacterial mechanisms, we subsequently focus on specific metal ions such as silver, zinc, copper, iron and gold, and outline their distinct modes of action. Finally, we discuss the use of these metal ions and nanoparticles in tissue engineering to prevent implant failure., M.G-G and U.E have received funding from the postdoctoral fellowship programme Beatriu de Pinós, funded by the Secretary of Universities and Research (Government of Catalonia) and by the Horizon 2020 programme of research and innovation of the European Union under the Marie Sklodowska-Curie grant agreement No 801370. R.A.P is supported by the Spanish Ministry by the Ramón y Cajal Program (RYC2018-025977-I). Additional financial support was provided by the Government of Catalonia (2017 SGR 708) and MINECO/FEDER project (RTI2018-096088-J-100).

[spa] En las últimas décadas la resistencia a antibióticos se ha convertido en uno de los principales problemas de salud pública a nivel global, y desde la iniciativa One-Health se han propuesto afrontar este problema, fundamentado en que la interconexión existente entre los biomas humanos, animales y ambientales favorece una mayor diseminación de las resistencias a antibióticos. Conocer las vías de movilización y transmisión de resistencias entre las diversas comunidades bacterianas es esencial para mitigar su dispersión. En los últimos años se ha incrementado el interés por el papel de los bacteriófagos o fagos como potenciales vectores de transferencia horizontal de genes entre bacterias (transducción), y en particular de genes de resistencia a antibióticos (GRAs). Su abundancia, persistencia en el ambiente y ubicuidad los hace perfectos vectores para la transducción de GRAs, y se empieza a considerar la transducción un evento de mayor relevancia en la movilización genética horizontal entre bacterias de lo que inicialmente se pensaba. A lo largo de los cinco capítulos en los que se ha estructurado esta tesis se profundiza en el papel de los fagos en la transmisión de resistencias, a través del estudio de la abundancia, diversidad y origen de partículas fágicas portadoras de GRAs en biomas humanos, animales y ambientales, siendo los alimentos los grandes protagonistas, debido a que la cadena alimentaria constituye un nexo entre dichos biomas. De los resultados obtenidos se puede concluir que a través de la dieta, las partículas fágicas pueden entrar en organismos humanos y animales e incrementar el número y diversidad de partículas fágicas portadoras de GRAs en el tracto intestinal, desde donde se podrían diseminar a otros órganos y transducir su material genético al entrar en contacto con algún huésped bacteriano sensible. Las partículas fágicas que contienen GRAs pasan, a su vez, al ambiente a través de los desechos humanos y animales, pudiendo contaminar de nuevo los alimentos. Se constituye, así, una via circular de entrada y salida de partículas fágicas y GRAs siguiendo una ruta oral-fecal-ambiente-oral. Ante los resultados obtenidos, y la ausencia de normativas en materia de seguridad alimentaria y ambiental que controlen los GRAs o los elementos genéticos que los movilizan, sería necesaria la identificación y monitoreo de las partículas fágicas portadoras de GRAs para evaluar si pueden constituir un riesgo para la salud humana y su contribución a la emergencia global de las resistencias a antibióticos., [eng] In recent decades, antibiotic resistance has become one of the biggest threats to global health. The One-Health approach has decided to tackle this problem, since the interconnection between human, animal and environmental biomes may favour the spreading of antibiotic resistances. Understanding the pathways of mobilization and transmission of resistances between different bacterial communities is crucial to mitigate its spread. In the recent years, there has been an increasing interest in the role of bacteriophages (or phages) as potential vectors for horizontal gene transfer of genes between bacteria through transduction, particularly in the mobilization of antibiotic resistance genes (ARGs). The ubiquity of phages makes the transduction of ARGs a more relevant event in genetic mobilization between bacteria than initially thought. The role of phages in the transmission of resistance is studied throughout the five chapters in which this PhD thesis has been structured, especially the study of the diversity of phage particles carrying ARGs in human, animal, and environmental biomes, focusing on food, because the food chain constitutes a nexus between these biomes. The results obtained allow to conclude through the diet, phage particles can enter human and animal organisms and increase the number and diversity of phage particles carrying ARGs in the intestinal tract, from where they may spread to other human and animal organs and transduce their genetic material upon contact with a sensitive bacterial host. Phage particles containing ARGs could end into the environment through human and animal waste, contaminating these biomes, following the oral-faecal-environment-oral route. Given the results obtained and the absence of regulations on food and environmental safety that control the ARGs or the genetic elements that mobilize them, it would be necessary to identify and monitor the page particles carrying ARGs to assess whether they may constitute a risk to human health and their contribution to the global emergence of antibiotic resistance.

Antiepileptic drug; Antiseizure medication; Tolerability Fármaco antiepiléptico; Medicamento anticonvulsivo; Tolerabilidad Medicament antiepilèptic; Medicament anticonvulsiu; Tolerabilitat Objective To characterize efficacy, safety/tolerability, and pharmacokinetics of padsevonil (PSL) administered concomitantly with ≤3 antiseizure medications (ASMs) for observable focal seizures in adults with drug-resistant epilepsy in two multicenter, randomized, double-blind, placebo-controlled, parallel-group trials. Methods The phase 2b dose-finding trial (EP0091/NCT03373383) randomized patients 1:1:1:1:1 to PSL 50/100/200/400 mg or placebo twice daily (b.i.d.). The phase 3 efficacy trial (EP0092/NCT03739840) randomized patients 1:1:1:1 to PSL 100/200/400 mg or placebo b.i.d. Patients with observable (focal aware with motor symptoms, focal impaired awareness, focal to bilateral tonic–clonic) focal seizures for ≥3 years, experiencing them ≥4 times per 28 days including during the 4-week baseline period despite treatment with ≥4 lifetime ASMs including current ASMs, were enrolled. Results In EP0091 and EP0092, 410 and 231 patients, respectively, were randomized and received at least one dose of trial medication. In patients in EP0091 on PSL 50/100/200/400 mg b.i.d. (n = 80/82/81/81, respectively) versus placebo (n = 81), outcomes included percentage reductions over placebo in observable focal seizure frequency during the 12-week maintenance period: 17.2%, 19.1% (p = 0.128), 19.2% (p = 0.128), 12.4% (p = 0.248); 75% responder rates (p-values for odds ratios): 13.8%, 12.2% (p = 0.192), 11.1% (p = 0.192), 16.0% (p = 0.124) versus 6.2%; 50% responder rates: 33.8% (p = 0.045), 31.7% (p = 0.079), 25.9% (p = 0.338), 32.1% (p = 0.087), versus 21.0%; TEAEs were reported by 82.7% (67/81), 78.3% (65/83), 74.4% (61/82), 90.1% (73/81) versus 78.3% (65/83). In patients in EP0092 on PSL 100/200/400 mg b.i.d. (n = 60/56/56, respectively) versus placebo (n = 54), outcomes included percentage reductions over placebo: −5.6% (p = 0.687), 6.5% (p = 0.687), 6.3% (p = 0.687); 75% responder rates: 15.3% (p = 0.989), 12.5% (p = 0.989), 14.3% (p = 0.989) versus 13.0%; 50% responder rates: 35.6% (p = 0.425), 33.9% (p = 0.625), and 42.9% (p = 0.125) versus 27.8%; TEAEs were reported by 80.0% (48/60), 78.9% (45/57), 83.1% (49/59) versus 67.3% (37/55). Significance In both trials, the primary outcomes did not reach statistical significance in any PSL dose group compared with placebo. PSL was generally well tolerated, and no new safety signals were identified.

The PI3K pathway is commonly activated in breast cancer, with PI3K-AKT pathway inhibitors used clinically. However, mechanisms that limit or enhance the therapeutic effects of PI3K-AKT inhibitors are poorly understood at a genome-wide level. Parallel CRISPR screens in 3 PTEN-null breast cancer cell lines identified genes mediating resistance to capivasertib (AKT inhibitor) and AZD8186 (PI3Kβ inhibitor). The dominant mechanism causing resistance is reactivated PI3K-AKT-mTOR signalling, but not other canonical signalling pathways. Deletion of TSC1/2 conferred resistance to PI3Kβi and AKTi through mTORC1. However, deletion of PIK3R2 and INPPL1 drove specific PI3Kβi resistance through AKT. Conversely deletion of PIK3CA, ERBB2, ERBB3 increased PI3Kβi sensitivity while modulation of RRAGC, LAMTOR1, LAMTOR4 increased AKTi sensitivity. Significantly, we found that Mcl-1 loss enhanced response through rapid apoptosis induction with AKTi and PI3Kβi in both sensitive and drug resistant TSC1/2 null cells. The combination effect was BAK but not BAX dependent. The Mcl-1i + PI3Kβ/AKTi combination was effective across a panel of breast cancer cell lines with PIK3CA and PTEN mutations, and delivered increased anti-tumor benefit in vivo. This study demonstrates that different resistance drivers to PI3Kβi and AKTi converge to reactivate PI3K-AKT or mTOR signalling and combined inhibition of Mcl-1 and PI3K-AKT has potential as a treatment strategy for PI3Kβi/AKTi sensitive and resistant breast tumours.

CDK4/6 inhibitors combined with endocrine therapy have demonstrated higher antitumor activity than endocrine therapy alone for the treatment of advanced estrogen receptor-positive breast cancer. Some of these tumors are de novo resistant to CDK4/6 inhibitors and others develop acquired resistance. Here, we show that p16 overexpression is associated with reduced antitumor activity of CDK4/6 inhibitors in patient-derived xenografts (n = 37) and estrogen receptor-positive breast cancer cell lines, as well as reduced response of early and advanced breast cancer patients to CDK4/6 inhibitors (n = 89). We also identified heterozygous RB1 loss as biomarker of acquired resistance and poor clinical outcome. Combination of the CDK4/6 inhibitor ribociclib with the PI3K inhibitor alpelisib showed antitumor activity in estrogen receptor-positive non-basal-like breast cancer patient-derived xenografts, independently of PIK3CA, ESR1 or RB1 mutation, also in drug de-escalation experiments or omitting endocrine therapy. Our results offer insights into predicting primary/acquired resistance to CDK4/6 inhibitors and post-progression therapeutic strategies. This study has been supported by the Susan G. Komen Foundation (CCR15330331) and by the Catalan Agency AGAUR (2017 SGR 540) [to V.S.]. V.S. received funds from the Instituto de Salud Carlos III: grants PI13/01714, CP14/00228, MV15/00041, CPII19/00033 and PI20/00892. M.P. received a Juan de la Cierva Grant from the Ministerio de Economía y Competitividad (FJCI-2015-25412), L.Mo. a grant from FI-AGAUR (2019 FI_B 01199), F.B-M. a grant from the Fundación Científica Asociación Española Contra el Cáncer (AECC_Postdoctoral17-1062) and M.S-G, a Marie Slodowska-Curie Innovative Training Networks PhD fellowship (H2020-MSCA-ITN-2015_675392). This work was supported by Breast Cancer Research Foundation (BCRF-19-08), Instituto de Salud Carlos III Project Reference number AC15/00062 and the EC under the framework of the ERA-NET TRANSCAN-2 initiative co-financed by FEDER, Instituto de Salud Carlos III (CB16/12/00449 and PI19/01181) and Asociación Española Contra el Cáncer (to J.A.). R.B.C. laboratory is supported by Breast Cancer Now (grant numbers: MAN-Q1 and MAN-Q2), NIHR Manchester Biomedical Research Centre (IS-BRC-1215-20007) and EdiREX Horizon 2020 grant No.731105. The xenograft program in the C.C. laboratory is supported by Cancer Research UK and also received funding from an EU H2020 Network of Excellence (EuroCAN). This work has been supported by NIH grants P30 CA008748 and RO1CA190642-01, the CDMRP grant BC171535P1, and the Breast Cancer Research Foundation [to M.S.]. A.P. received funds from Instituto de Salud Carlos III—PI16/00904 and PI19/01846, Breast Cancer Now—2018NOVPCC1294, Breast Cancer Research Foundation-AACR Career Development Awards for Translational Breast Cancer Research 19-20-26-PRAT, Fundació La Marató TV3 201935-30, the European Union’s Horizon 2020 research and innovation program H2020-SC1-BHC-2018-2020. IRB Barcelona is a recipient of a Severo Ochoa Centre of Excellence Award from the Spanish Ministry of Economy and Competitiveness (MINECO; Government of Spain) and is supported by CERCA (Generalitat de Catalunya). C. S. Verma reports grants from MSD International and grants from Ipsen outside the submitted work.

Candida parapsilosis; Antifungal resistance; Outbreaks Candida parapsilosis; Resistencia antifúngica; Brotes Candida parapsilosis; Resistència antifúngica; Brots Background Candida parapsilosis is a frequent cause of candidemia worldwide. Its incidence is associated with the use of medical implants, such as central venous catheters or parenteral nutrition. This species has reduced susceptibility to echinocandins, and it is susceptible to polyenes and azoles. Multiple outbreaks caused by fluconazole-nonsusceptible strains have been reported recently. A similar trend has been observed among the C. parapsilosis isolates received in the last 2 years at the Spanish Mycology Reference Laboratory. Methods Yeast were identified by molecular biology, and antifungal susceptibility testing was performed using the European Committee on Antimicrobial Susceptibility Testing protocol. The ERG11 gene was sequenced to identify resistance mechanisms, and strain typing was carried out by microsatellite analysis. Results We examined the susceptibility profile of 1315 C. parapsilosis isolates available at our reference laboratory between 2000 and 2021, noticing an increase in the number of isolates with acquired resistance to fluconazole, and voriconazole has increased in at least 8 different Spanish hospitals in 2020–2021. From 121 recorded clones, 3 were identified as the most prevalent in Spain (clone 10 in Catalonia and clone 96 in Castilla-Leon and Madrid, whereas clone 67 was found in 2 geographically unrelated regions, Cantabria and the Balearic Islands). Conclusions Our data suggest that concurrently with the coronavirus disease 2019 pandemic, a selection of fluconazole-resistant C. parapsilosis isolates has occurred in Spain, and the expansion of specific clones has been noted across centers. Further research is needed to determine the factors that underlie the successful expansion of these clones and their potential genetic relatedness. O.Z. was funded by grants SAF2017–86912-R and PID2020–114546RB-I00 from the Spanish Ministry for Science and Innovation. This work was also funded by the National Centre for Microbiology (Instituto de Salud Carlos III) through the Surveillance Program of Antifungal Resistance and the Center for Biomedical Research in Network of Infectious Diseases CIBERINFECTCB21/13/00105 (O.Z. and L.A.F.), CIBERINFEC-CB21/13/00009 (M.P.-A.), CIBERES-CB06/06/0037 (C.A.-T.), and CIBERES-CB06/06/0058 (J.G). L.A.-F. was supported by Fondo de Investigación Sanitaria (MPY 117/18 and MPY 305/20). We thank Dr. David Campany Herrero (Vall d’Hebron Hospital), Noelia Garrido Peño (Móstoles Hospital), David Gómez Gómez y Aitziber Illaro Uranga (Marqués de Valdecilla Hospital), María Ángeles Machín Morón (Burgos Hospital), Jose Manuel Caro Teller (Doce de Octubre Hospital), Marina Calvo (Puerta de Hierro Hospital), and Ariadna Padulles (Bellvitge Hospital) for providing the data on antifungal consumption from their hospitals. We also thank Ángel Zaballos and Pilar Jiménez from the Genomics Core Facility from Instituto de Salud Carlos III for their technical help with the microsatellite analysis technique.

Medicamentos anticonvulsivos; Cenobamato; Eficiencia Medicaments anticonvulsius; Cenobamat; Eficiència Anti-seizure medications; Cenobamate; Efficiency Introduction Epilepsy is a serious neurological disease, ranking high in the top causes of disability. The main goal of its treatment is to achieve seizure freedom without intolerable adverse effects. However, approximately 40% of patients suffer from Drug-Resistant Epilepsy (DRE) despite the availability of the latest options called third-generation Anti-Seizure Medications(ASMs). Cenobamate is the first ASM approved in Spain for the adjunctive treatment of Focal-Onset Seizures (FOS) in adult patients with DRE. The introduction of a new drug increases the number of therapeutic options available, making it important to compare it with existing alternatives in terms of clinical benefit and efficiency. Purpose This study aimed to compare the clinical benefit, in terms of the Number Needed to Treat (NNT), and the efficiency, in terms of Cost per NNT (CNT), associated with cenobamate versus third-generation ASMs used in Spain for the adjunctive treatment of FOS in patients with DRE. Methods The Number Needed to Treat data was calculated based on the ≥50% responder rate and seizure freedom endpoints (defined as the percentage of patients achieving 50% and 100% reduction in seizure frequency, respectively), obtained from pivotal clinical trials performed with cenobamate, brivaracetam, perampanel, lacosamide, and eslicarbazepine acetate. The NNT was established as the inverse of the treatment responder rate minus the placebo responder rate and was calculated based on the minimum, mid-range Daily Defined Dose (DDD), and maximum doses studied in the pivotal clinical trials of each ASM. CNT was calculated by multiplying the annual treatment cost by NNT values for each treatment option. Results In terms of NNT, cenobamate was the ASM associated with the lowest values at all doses for both ≥50% responder rate and seizure freedom compared with the alternatives. In terms of CNT, for ≥50% responder rate, cenobamate was the ASM associated with the lowest CNT values at DDD and lacosamide and eslicarbazepine acetate at the minimum and maximum dose, respectively. For seizure freedom, cenobamate was associated with the lowest CNT value at DDD and the maximum dose and lacosamide at the minimum dose. Conclusions Cenobamate could represent the most effective ASM in all doses studied compared to the third-generation ASMs and the most efficient option at DDD for both ≥50% responder rate and seizure freedom. This study could represent an important contribution towards informed decision-making regarding the selection of the most appropriate therapy for FOS in adult patients with DRE from a clinical and economical perspective in Spain. This study was sponsored by Angelini Pharma España, S.L.U.

[eng] The main objectives of this thesis are to study the opportunistic infection caused by human cytomegalovirus and SARS-CoV-2 inquiring persistent infection and antiviral resistance. These major objectives can be further divided into: 1. Characterization of mutations associated with antiviral resistance in immunocompromised patients with refractory human cytomegalovirus infection. 1.1. Genotypic antiviral resistance testing of human cytomegalovirus target genes of conventional and new antiviral therapies by Sanger and next generation sequencing. 1.2. Phenotypic studies of uncharacterized mutations found genotypically in clinical samples by bacmid technologies, to determine the antiviral susceptibility and the replicative capacity of each individual mutation. 1.3. Searching for baseline resistant mutations before the administration of new therapies to prevent antiviral treatment failure. 1.4. Determining the incidence of cytomegalovirus antiviral resistance mutations, natural polymorphisms, and uncharacterized genetic variants in immunocompromised patients with clinically resistant cytomegalovirus infection. 2. Quantification of SARS-CoV-2 normalized viral loads in respiratory samples to study the dynamics of total viral RNA. 3. Determination of SARS-CoV-2 replicative capacity during the infection course by the presence of subgenomic RNA, and its broad applicability on the patients’ clinical monitoring. 4. Assessment of patients with persistent SARS-CoV-2 replication and/or severe COVID-19 treated with remdesivir. 5. Search of mutations associated with remdesivir failure by next-generation sequencing in severe COVID-19 patients., [spa] Esta tesis estudia la caracterización genotípica y fenotípica de mutaciones de resistencia a los anvitivirales, asi como la replicación viral persistente, en el paciente inmunocomprometido. En concreto se estudia la infección por citomegalovirus por su alta prevalencia en la población y por su alta mortalidad en pacientes imunocomprometidos, y la infección emergente por SARS-CoV-2 debido a su rápida diseminación causante de la pandemia COVID-19, la cual ha afectado especialmente a este tipo de paciente condicionando a una replicación prolongada, al uso de fármacos antivirales y la aparición de resistencias.

Antibiotic lock technique; Biofilms; Catheter-related infection Técnica de bloqueo antibiótico; Biopelículas; Infección relacionada con el catéter Tècnica de bloqueig antibiòtic; Biofilms; Infecció relacionada amb el catèter The aims of this study were as follows. First, we determined the antimicrobial efficacy of hypochlorous acid (HClO) against bacterial, fungal, and yeast strains growing planktonically and growing in biofilms. Second, we sought to compare the activity of the combination of daptomycin and HClO versus those of the antimicrobial agents alone for the treatment of experimental catheter-related Staphylococcus epidermidis infection (CRI) using the antibiotic lock technique (ALT) in a rabbit model. HClO was generated through direct electric current (DC) shots at determined amperages and times. For planktonic susceptibility studies, 1 to 3 DC shots of 2, 5, and 10 mA from 0 to 300 s were applied. A DC shot of 20 mA from 0 to 20 min was applied to biofilm-producing strains. Central venous catheters were inserted into New Zealand White rabbits, inoculated with an S. epidermidis strain, and treated with saline solution or ALT using daptomycin (50 mg/mL), HClO (20 mA for 45 min), or daptomycin plus HClO. One hundred percent of the planktonic bacterial, fungal, and yeast strains were killed by applying one DC shot of 2, 5, and 10 mA, respectively. One DC shot of 20 mA for 20 min was sufficient to eradicate 100% of the tested biofilm-producing strains. Daptomycin plus HClO lock therapy showed the highest activity for experimental CRI with S. epidermidis. HClO could be an effective strategy for treating infections caused by extensively drug-resistant or multidrug-resistant and biofilm-producing strains in medical devices and chronic wounds. The results of the ALT using daptomycin plus HClO may be promising. IMPORTANCE Currently, drug-resistant infections are increasing and there are fewer antibiotics available to treat them. Therefore, there is an urgent need to find new antibiotics and nonantimicrobial strategies to treat these infections. We present a new nonantibiotic strategy based on hypochlorous acid generation to treat long-term catheter-related and chronic wounds infections. This study was supported by research grants from the Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III (FIS 01162), la Marató TV3 (472/U/2018), and CaixaImpulse Program (Fundació “LaCaixa”) and the Spanish Network for the Research in Infectious Diseases (REIPI RD19/0016).

Streptococcal infections are usually treated with beta-lactam antibiotics, but, in case of allergic patients or reduced antibiotic susceptibility, macrolides and fluoroquinolones are the main alternatives. This work focuses on studying macrolide resistance rates, genetic associated determinants and antibiotic consumption data in Spain, Europe and also on a global scale. Macrolide resistance (MR) determinants, such as ribosomal methylases (erm(B), erm(TR), erm(T)) or active antibiotic efflux pumps and ribosomal protectors (mef(A/E)-mrs(D)), are differently distributed worldwide and associated with different clonal lineages and mobile genetic elements. MR rates vary together depending on clonal dynamics and on antibiotic consumption applying selective pressure. Among Streptococcus, higher MR rates are found in the viridans group, Streptococcus pneumoniae and Streptococcus agalactiae, and lower MR rates are described in Streptococcus pyogenes. When considering different geographic areas, higher resistance rates are usually found in East-Asian countries and milder or lower in the US and Europe. Unfortunately, the availability of data varies also between countries; it is scarce in low- and middle- income countries from Africa and South America. Thus, surveillance studies of macrolide resistance rates and the resistance determinants involved should be promoted to complete global knowledge among macrolide resistance dynamics.

Background Despite atezolizumab and bevacizumab (A + B) is currently the first‐line treatment for hepatocellular carcinoma (HCC) patients, some patients will not be adequate for this combination. In the setting of sorafenib some adverse events have been proposed as prognostic factors. Objective To characterize the early diarrhoea development as prognostic factor in 344 HCC patients. Methods The development of early diarrhoea in sorafenib treatment defined as patients who developed diarrhoea and needed dose modification within the first 60 days of treatment (e‐diarrhoea) and 3‐grouping variables were analysed: Patients with e‐diarrhoea, patients who developed diarrhoea after the first 60 days of treatment (L‐diarrhoea) and patients that never developed diarrhoea (never diarrhoea). Results The median overall survival in sorafenib treated patients was significantly different across groups (6.8 months for e‐diarrhoea, 26.7 months for L‐diarrhoea and 13.3 months for never‐diarrhoea). The emergence of e‐diarrhoea was associated with poor outcomes (hazard ratio [HR] 1.84 [95%CI 1.15–2.95]), while there was no increased/decreased risk of dismal evolution in patients with L‐diarrhoea (HR 0.66 [95%CI 0.42–1.03]). Conclusion The emergence of e‐diarrhoea in HCC patients treated with sorafenib is an early predictor of dismal evolution under this therapy. Thus, prompt identification of these non‐responders may be useful for an early switch to second‐line therapies., Key Summary Established knowledge on this subject Diarrhoea is a frequent adverse event of sorafenib and its emergence has been associated to better outcomes. What are the significant and/or new findings of this study? Early diarrhoea (e‐diarrhoea) in hepatocellular carcinoma patients treated with sorafenib is an early predictor of dismal evolution.Diarrhoea under sorafenib should not be taken as a predictive parameter of lack of benefit.E‐diarrhoea could be used as clinical biomarker for switching to second‐line therapies.

Resistència antibiòtica; Salmonel·la no tifòdica; Salmonel·la tífica Resistencia antibiótica; Salmonela no tifódica; Salmonela tífica Antibiotic resistance; Non-typhoid Salmonella; Typhus Salmonella Aquest informe té com a objectiu analitzar les característiques epidemiològiques dels casos confirmats de salmonel·la no tifòdica i salmonel·la tífica i analitzar la sensibilitat antimicrobiana dels casos declarats a l’SNMC durant els anys 2016-2019.

[cat] INTRODUCCIÓ: Aproximadament, un 40% dels pacients amb epilèpsia focal son fàrmac-resistents. En aquests casos, la cirurgia de l’epilèpsia pot controlar o reduir significativament les crisis. La zona epileptògena (ZE) és la regió cerebral que la seva resecció implicaria la llibertat de crisis. Malauradament, no hi ha cap prova que determini amb precisió la ZE. HIPÒTESIS: Les alteracions de la DTI que valora la integritat de la substància blanca i les anomalies corticals determinades per la PET-FDG estan relacionades. Per altra banda, el PISCOM té una utilitat similar al SISCOM per detectar la zona d’inici ictal, però evitant la realització de la SPECT interictal. Aquests dos nous processaments de la neuroimatge cerebral permetrien determinar amb major precisió la ZE. OBJECTIUS: Valorar la utilitat de la combinació de la DTI amb la PET-FDG i el PISCOM a l’avaluació pre-quirúrgica de l’epilèpsia fàrmac-resistent. De forma més específica, es pretén determinar la localització de les alteracions de la DTI i la seva relació amb l’hipometabolisme cortical de la PET- FDG i la resecció de la potencial ZE, en pacient adults amb epilèpsia temporal medial amb esclerosi de hipocamp (ETM + EH). En el cas del PISCOM, l’objectiu es valorar la validesa del PISCOM per a identificar la potencial ZE, comparat amb el SISCOM ¡ la PET-FDG, en pacients pediàtrics amb una epilèpsia focal fàrmac-resistent. MÈTODES: De forma retrospectiva, els resultats de la DTI i la PET-FDG en una mostra de 21 pacients adults amb ETM + EH es van comparar amb un grup control sa, i a 18 d’aquests pacients se’ls havia realitzat una cirurgia d’epilèpsia. Respecte al PISCOM, la mostra estava formada per 22 pacients pediàtrics amb epilèpsia focal fàrmac-resistent, majoritàriament extra-temporal. Els resultats del PISCOM es van comparar amb les alteracions determinades pel SISCOM i la PET- FDG, realitzades durant l’avaluació pre-quirúrgica, i amb la ZE determinada per la resecció quirúrgica, la histologia, i el seguiment clínic post-quirúrgic favorable. RESULTATS: Les anomalies determinades per la combinació de la DTI i la PET-FDG van ser extenses, però sent més difuses quan la ETM + EH estava lateralitzada a l’hemisferi esquerre. Segons l’extensió de l’hipometabolisme de la PET-FDG al lòbul temporal epilèptic, es van diferenciar tres grups: anterior, mig i posterior. El grup anterior es relacionava amb alteracions de la DTI més anteriors, a diferència dels altes dos grups on les anomalies de la DTI eren tant anteriors com posteriors. No es va objectivar una relació clara entre les troballes de la DTI i la PET-FDG, i els resultats post-quirúrgics. El PISCOM va localitzar de forma exitosa la ZE en un major número de casos que el SISCOM (86% vs 54%, respectivament), i en una proporció similar a la PET-FDG. La ZE va ser identificada en tots els pacients només sent necessaris els resultats de la PET-FDG i el PISCOM. Es va observar una major concordança global entre observadors a l’analitzar els resultats del PISCOM, comparats amb els de la PET-FDG i el SISCOM. CONCLUSIONS: Els resultats de la combinació de la DTI i la PET-FDG mostren una relació entre ambdues tècniques, permetent identificar diferents xarxes neuronals implicades en una mateixa patologia (ETM + EH), en funció de l’extensió de l’hipometabolisme de la PET-FDG, però sense una clara relació amb el resultats post-quirúrgics. Els resultats del PISCOM suggereixen que podria ser una tècnica complementària per inferir la ZE als casos amb una PET-FDG dubtosa o negativa, substituint al SISCOM i evitant la realització de la SPECT interictal. Ambdues noves tècniques de neuroimatge podrien tenir una utilitat a l’avaluació pre-quirúrgica de l’epilèpsia per ajudar a inferir de forma més precisa la localització de la ZE., [eng] PURPOSE: In drug-resistant epilepsy, surgery can control or significantly reduce seizures. The epileptogenic zone (EZ) is the brain region whose resection would imply freedom from seizures. Unfortunately, there is no evidence to accurately determine EZ. The aim of this study is to determine whether the extent of 18F-FDG-PET hypometabolism combined to DTI abnormalities, and the PISCOM, a new nuclear imaging processing technique, could be useful to try to locate the EZ. METHODS: Twenty-one patients with MTLE-HS underwent comprehensive preoperative evaluation; 18 (86%) of these underwent epilepsy surgery. We analyzed and compared the pattern of white matter (WM) alterations on DTI and cortical hypometabolism on 18F-FDG-PET. Twenty-two children with pharmacorefractory epilepsy, mainly extratemporal, who had undergone presurgical assessment including SISCOM and 18F-FDG-PET and with postsurgical favorable outcome (Engel class I or II) for at least two years, were included. RESULTS: Widespread temporal and extratemporal 18F-FDG-PET and DTI abnormalities were found, involved similar brain regions, being more extensive in the left than the right MTLE-HS. Three groups were determined according to temporal 18F-FDG-PET patterns: hypometabolism restricted to the anterior third, to the middle third, and extending to the posterior third. Patients with anterior temporal hypometabolism showed DTI abnormalities in anterior tracts while patients with posterior hypometabolism showed WM alterations in anterior and posterior tracts. PISCOM concordance with the known EZ was significantly higher than SISCOM (p

Virologic failure; Hepatitis C Fracaso virológico; Hepatitis C Fracàs virològic; Hepatitis C Background & Aims Direct-acting antiviral (DAA) regimens provide a cure in >95% of patients with chronic HCV infection. However, in some patients in whom therapy fails, resistance-associated substitutions (RASs) can develop, limiting retreatment options and risking onward resistant virus transmission. In this study, we evaluated RAS prevalence and distribution, including novel NS5A RASs and clinical factors associated with RAS selection, among patients who experienced DAA treatment failure. Methods SHARED is an international consortium of clinicians and scientists studying HCV drug resistance. HCV sequence linked metadata from 3,355 patients were collected from 22 countries. NS3, NS5A, and NS5B RASs in virologic failures, including novel NS5A substitutions, were examined. Associations of clinical and demographic characteristics with RAS selection were investigated. Results The frequency of RASs increased from its natural prevalence following DAA exposure: 37% to 60% in NS3, 29% to 80% in NS5A, 15% to 22% in NS5B for sofosbuvir, and 24% to 37% in NS5B for dasabuvir. Among 730 virologic failures, most were treated with first-generation DAAs, 94% had drug resistance in ≥1 DAA class: 31% single-class resistance, 42% dual-class resistance (predominantly against protease and NS5A inhibitors), and 21% triple-class resistance. Distinct patterns containing ≥2 highly resistant RASs were common. New potential NS5A RASs and adaptive changes were identified in genotypes 1a, 3, and 4. Following DAA failure, RAS selection was more frequent in older people with cirrhosis and those infected with genotypes 1b and 4. Conclusions Drug resistance in HCV is frequent after DAA treatment failure. Previously unrecognized substitutions continue to emerge and remain uncharacterized. The initial SHARED development was funded in part by Merck and a User Partnership Program grant from Genome British Columbia to P.R.H and A.Y.M.H (UPP029). The Kirby Institute is funded by the Australian Government Department of Health and Ageing. The views expressed in this publication do not necessarily represent the position of the Australian (or any other) Government. C.R. and J.G. are supported by an NHMRC Investigator Grant (nos. 1173666, 1176131). An NHMRC Program Grant supported RAS testing and data collection by M.W.D. (1053206) and small grants from the Australian Centre for HIV and Hepatitis Virology Research, The University of Sydney, Western Sydney Local Health District Research Education Network, and the Robert W. Storr bequest to the Sydney Medical Foundation (University of Sydney). S.P. received personal fees from Gilead Sciences. Regarding the Italian data, the work was supported in part by the Italian Ministry of Instruction, University and Research (MIUR) (Bandiera InterOmics Protocollo PB05 1°), by the Italian Ministry of Health (RF-2016-02362422), and by Aviralia and Vironet C Foundations.

Breast cancer; Drug tolerance; Secretome Cáncer de mama; Tolerancia a los fármacos; Secretoma Càncer de mama; Tolerància als fàrmacs; Secretoma Drug tolerant persister (DTP) cells enter into a reversible slow-cycling state after drug treatment. We performed proteomic characterization of the breast cancer (BC) DTP cell secretome after eribulin treatment. We showed that the growth differentiation factor 15 (GDF15) is a protein significantly over-secreted upon eribulin treatment. The biomarker potential of GDF15 was confirmed in 3D-cell culture models using BC cells lines and PDXs, as well as in a TNBC in vivo model. We also found that GDF15 is required for survival of DTP cells. Direct participation of GDF15 and its receptor GFRAL in eribulin-induction of DTPs was established by the enhanced cell killing of DTPs by eribulin seen under GDF15 and GFRAL loss of function assays. Finally, we showed that combination therapy of eribulin plus an anti-GDF15 antibody kills BC-DTP cells. Our results suggest that targeting GDF15 may help eradicate DTP cells and block the onset of acquired resistance. This research was funded by Eisai Inc.

Campylobacter jejuni is a foodborne pathogen causing bacterial gastroenteritis, with the highest incidence reported in Europe. The prevalence of antibiotic resistance in C. jejuni, as well as in many other bacterial pathogens, has increased over the last few years. In this report, we describe the presence of a plasmid in a multi-drug-resistant C. jejuni strain isolated from a gastroenteritis patient. Mating experiments demonstrated the transference of this genetic element (pCjH01) among C. jejuni by plasmid conjugation. The pCjH01 plasmid was sequenced and assembled, revealing high similarity (97% identity) with pTet, a described tetracycline resistance encoding plasmid. pCjH01 (47.7 kb) is a mosaic plasmid composed of a pTet backbone that has acquired two discrete DNA regions. Remarkably, one of the acquired sequences carried an undescribed variant of the aadE-sat4-aphA-3 gene cluster, providing resistance to at least kanamycin and gentamycin. Aside from the antibiotic resistance genes, the cluster also carries genes coding for putative regulators, such as a sigma factor of the RNA polymerase and an antisigma factor. Homology searches suggest that Campylobacter exchanges genetic material with distant G-positive bacterial genera. This research was funded by the Spanish Ministry of Economy and Competitiveness [grant AGL2013-45339R], the Spanish Ministry of Science, Innovation and Universities (MCIU), State Bureau of Investigation (AIE), the European Regional Development Fund (FEDER) [grant PGC2018-096958-B-I00], and the Catalonian government [grant 2017SGR499]. PG was the recipient of an ADR fellowship of the University of Barcelona

[eng] Increased expression of TIMP-1 is associated with poor prognosis in virtually all cancer types, including lung cancer. However, how TIMP-1 is regulated in lung cancer and how it drives tumor progression is poorly defined. In the first part of this thesis, we analyze the expression of TIMP-1 and its cell surface receptor CD63 in two major lung cancer subtypes: lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC). We report that TIMP-1 is aberrantly overproduced in tumor-associated fibroblasts (TAFs) in ADC compared to SCC-TAFs, and identify the selective hyperactivity of the pro-fibrotic TGF- β1/SMAD3 pathway in ADC-TAFs as a major mechanism driving the excessive secretion of TIMP-1 in ADC-TAFs. In contrast, CD63 was markedly downregulated in SCC cells compared to ADC cells. In the second part of this thesis, we define the tumor-promoting effects of the heterotypic crosstalk, specific for ADC patients, between TIMP-1 secreted by TAFs and CD63 in cancer cells. Functional assays revealed that TIMP-1 within the pro- tumoral secretome of TGF-β1-activated ADC-TAFs is necessary and sufficient to promote the growth and invasion of ADC cells in culture. Concomitantly, tumor cell expression of CD63 was required for the tumor-promoting effects of TIMP-1. Consistently, in vivo analyses revealed that ADC cells co-injected with fibroblasts with reduced TIMP-1 expression into immunocompromised mice exhibited a lower tumor volume together with a less invasive growth pattern compared to tumors bearing parental fibroblasts. Finally, we studied the relationship of TIMP-1/CD63 crosstalk with the resistance mechanism of SCC patients to nintedanib, which is an antifibrotic drug that has been selectively approved for the treatment of ADC patients. Of note, nintedanib strongly inhibited TIMP-1 secretion in TAFs. Moreover, we observed that TIMP-1 deficient TAFs are resistant to the nintedanib inhibition of their pro-tumoral traits, both in vitro and in vivo, compared to parental TAFs. This work identifies the first tumor-promoting TAF- carcinoma crosstalk that is selective for lung ADC, provides new insights on the pathologic role of stromal TIMP-1 in lung cancer, identifies a novel resistance mechanism to nintedanib in SCC, and points to the TIMP-1/CD63 interaction as a novel therapeutic target., [cat] L’expressió elevada de TIMP-1 està associada a un mal pronòstic en gairebé tots els tipus de càncer, inclòs el càncer de pulmó. No obstant, es desconeix com es regula l’expressió de TIMP-1 en càncer de pulmó ni mitjançant quins mecanismes promou la progressió tumoral. En la primera part d’aquesta tesi, s’han analitzat els nivells de TIMP-1 i CD63, el seu receptor de membrana, en els dos subtipus majoritaris de càncer de pulmó: l’adenocarcinoma (ADC) i el carcinoma escatós (CE). Hem trobat que en els fibroblasts associats al tumor (TAFs de l’anglès) del subtipus ADC hi ha una secreció aberrant de TIMP-1 en comparació amb els TAFs de CE. Aquesta excessiva secreció de TIMP-1 en ADC es deu a la hiperactivació selectiva de la via del TGF-β1/SMAD3 en aquest subtipus de TAFs. D’altra banda, els nivells de CD63 són més baixos en les cèl·lules canceroses de CE que en les d’ADC. En la segona part d’aquesta tesi, hem descrit els efectes pro- tumorals d’un mecanisme de comunicació creuada entre el TIMP-1 secretat pels TAFs i el CD63 de les cèl·lules canceroses que és específic per als pacients amb ADC. El TIMP- 1 secretat pels TAFs d’ADC és necessari i suficient per promoure el creixement i la invasió en les cèl·lules de càncer. Simultàniament, l’expressió de CD63 en les cèl·lules tumorals es necessària per observar aquests efectes pro-tumorals del TIMP-1. De manera consistent, in vivo hem observat que la inhibició de la secreció del TIMP-1 redueix tant el volum tumoral com el creixement invasiu en ratolins co-injectats amb cèl·lules canceroses i TAFs. Finalment, hem estudiat la relació d’aquesta comunicació creuada amb la resistència al nintedanib trobada en pacients amb CE, el qual és un fàrmac antifibròtic aprovat selectivament en pacients amb ADC. Hem observat que en els TAFs en els quals s’ha inhibit genèticament la secreció de TIMP-1, hi ha una menor resposta a nintedanib. Aquest treball identifica per primer cop un mecanisme de comunicació creuada entre TAFs i cèl·lules canceroses que és específic per pacients amb ADC, aporta nous coneixements sobre el rol patològic del TIMP-1 en càncer de pulmó, descriu un nou mecanisme de resistència al nintedanib en CE i apunta la interacció TIMP-1/CD63 com a nova diana terapèutica.

The aim of this transversal study was to describe the virological and immunological features of HIV-infected youths transferred from pediatric to adult care units since 1997 vs. the non-transferred patients from the Madrid Cohort of HIV-infected children and adolescents in Spain. We included 106 non-transferred and 184 transferred patients under clinical follow-up in 17 public hospitals in Madrid by the end of December 2017. Virological and immunological outcomes were compared in transferred vs. non-transferred patients. ART drug resistance mutations and HIV-variants were analyzed in all subjects with available resistance pol genotypes and/or genotypic resistance profiles. Among the study cohort, 133 (72.3%) of 184 transferred and 75 (70.7%) of 106 non-transferred patients had available resistance genotypes. Most (88.9%) of transferred had ART experience at sampling. A third (33.3%) had had a triple-class experience. Acquired drug resistance (ADR) prevalence was significantly higher in pretreated transferred than non-transferred patients (71.8% vs. 44%; p = 0.0009), mainly to NRTI (72.8% vs. 31.1%; p

MDR; Tratamiento; Tuberculosis MDR; Tractament; Tuberculosi MDR; Treatment; Tuberculosis Multidrug-resistant (MDR) tuberculosis (TB), resistant to isoniazid and rifampicin, continues to be one of the most important threats to controlling the TB epidemic. Over the last few years, there have been promising pharmacological advances in the paradigm of MDR TB treatment: new and repurposed drugs have shown excellent bactericidal and sterilizing activity against Mycobacterium tuberculosis and several all-oral short regimens to treat MDR TB have shown promising results. The purpose of this comprehensive review is to summarize the most important drugs currently used to treat MDR TB, the recommended regimens to treat MDR TB, and we also summarize new insights into the treatment of patients with MDR TB. M.L.A. was supported by “Rio Hortega” (CM19/00070) grant from Instituto Carlos III through the Ministry of Economy and Competitiveness, Spain. A.S.M. was supported by a postdoctoral grant “Juan Rodés” (JE18/00022) from the Instituto de Salud Carlos through the Ministry of Economy and Competitiveness, Spain.

Este artículo contiene 8 páginas, 5 figuras., Antibiotic resistance has become a major Global Health concern and a better understanding on the global spread mechanisms of antibiotic resistant bacteria (ARB) and intercontinental ARB exchange is needed. We measured atmospheric depositions of antibiotic resistance genes (ARGs) by quantitative (q)PCR in rain/snow collected fortnightly along 4 y. at a remote high mountain LTER (Long-Term Ecological Research) site located above the atmospheric boundary layer (free troposphere). Bacterial composition was characterized by 16S rRNA gene sequencing, and air mass provenances were determined by modelled back trajectories and rain/snow chemical composition. We hypothesize that the free troposphere may act as permanent reservoir and vector for ARB and ARGs global dispersal. We aimed to i) determine whether ARGs are long-range intercontinental and persistently dispersed through aerosols, ii) assess ARGs long-term atmospheric deposition dynamics in a remote high mountain area, and iii) unveil potential diffuse ARGs pollution sources. We showed that the ARGs sul1 (resistance to sulfonamides), tetO (resistance to tetracyclines), and intI1 (a proxy for horizontal gene transfer and anthropogenic pollution) were long-range and persistently dispersed in free troposphere aerosols. Major depositions of tetracyclines resistance matched with intensification of African dust outbreaks. Potential ARB mostly traced their origin back into agricultural soils. Our study unveils that air masses pathways are shaping ARGs intercontinental dispersal and global spread of antibiotic resistances, with potential predictability for interannual variability and remote deposition rates. Because climate regulates aerosolization and long-range air masses movement patterns, we call for a more careful evaluation of the connections between land use, climate change and ARB long-range intercontinental dispersal., This research is included within the global surveillance activities of the Long-Term Ecological Research node Aigüestortes (LTER-AT) and was supported by Grant ECOSENSOR-BIOCON 04/009 from BBVA Foundation; Grants AERBAC 079/2007, AERBAC-2 178/2010, and DISPERSAL 829/2013 from the Spanish Office for the Environment’s- National Parks Research Programme (OAPN); and INTERACTOMA RTI2018-101205-B-I00 from the Spanish Agency of Research (AEIMICINN) and European funding (ERDF) to EOC. JC was supported by PTA2018-016527-I (AEI-MICINN) and XTM by grant INTERACTOMA. CB wish to thank the support from the Economy and Knowledge Department of the Catalan Government through Consolidated Research Group (ICRA-ENV 2017 SGR 1124).

[eng] Following the laws of evolution, bacteria are capable of slowly but inexorably acquiring resistance to antibiotic compounds that we have been using for decades. Therefore, bacteria that seemed harmless to most people (such as E. coli or P. aeruginosa) have become dangerous agents capable of alarming resilience. Furthermore, due to the ease in which bacteria develop new antibiotic resistances, or even share them across multiple species, the development of new compounds is no longer profitable for most pharma companies, which makes them not to prioritize this research field. These facts result in the non-existent development of new families of antibiotics, which supposes the risk of a future in which bacteria cannot be controlled without medication. In this context, many research groups have started working in the development of new antibiotics with more efficacy or less toxic effects than the ones that already exist. In this thesis, we focus on the study of an antimicrobial lipopeptide with effect against Gram-negative bacteria, colistin, and how to alter its composition or peptide sequence can potentially mean the development of new antimicrobial analogues that show a greater therapeutic profile. To do that, we used solid phase synthesis to generate the peptide analogues and we tested them for antimicrobial activity both in vitro and in vivo, and analysed their biophysical properties when exposing synthetic and natural membranes to them. Among other modifications, we can highlight the length of the fat chain of the lipopeptide, even with the substitution of the residues in position 6 and 7 without changing the net load. As a result of our research, we demonstrated that the solid phase synthesis of peptides is a consistent method for producing pure composts that allows industrial scale. Regarding the obtained peptide analogues, we confirm that their antibiotic action had a similar mechanism to the natural colistin. We also observed the differences between the activity of the analogues depending on the length of the chain of fatty acid. Also, the substitution of the fatty acid chain for other hydrophobic chains enhances the antimicrobial activity of the resulting peptides, allowing for a simpler and more efficient synthesis. Finally, the star peptide of thesis shows an antimicrobial capacity against gram-negative bacteria similar to colistin. Furthermore, it shows pharmacokinetic and nephrotoxic profiles potentially compatible with the potential administration to animals. Given that the effect of the star peptide in the treatment of a bacterial pneumonia model is not inferior to that of colistin, we consider that it is a promising compound which we would need to continue researching and developing so that one day it could reach the market., [cat] Seguint les conegudes lleis de l'evolució, els bacteris són capaços d'adquirir lenta però inexorablement resistència als compostos antibiòtics que fa dècades que fem servir. Com a conseqüència, bacteris que semblaven innocus per a la majoria de les persones (com E. coli o P. aeruginosa) s'han convertit en agents perillosos capaços d'una resiliència alarmant. Aquest fet, juntament amb el desenvolupament gairebé inexistent de noves famílies d'antibiòtics, suposa el risc d'un futur en què les infeccions bacterianes no puguin ser tractades eficaçment. En aquest context, nombrosos grups treballen en el desenvolupament de nous antibiòtics amb més eficàcia o menys efectes tòxics als ja existents. En aquesta tesi, ens centrem en l'estudi d'un lipopèptid antimicrobià amb activitat contra bacteris gramnegatius, la colistina, i de com alterar la seva estructura peptídica pot suposar el desenvolupament de nous anàlegs antimicrobians que potencialment mostrin un perfil terapèutic més gran. Entre altres modificacions, podem ressaltar la longitud de la cadena d'àcid gras del lipopèptid, així com la substitució dels residus en posició 6 i 7 sense canviar la càrrega neta. Com a resultat de la nostra investigació, demostrem que la síntesi en fase sòlida de pèptids és un mètode consistent per produir compostos purs que permet l'escalat industrial. Pel que fa als anàlegs obtinguts, comprovem que la seva acció antibiòtica té lloc seguint un mecanisme semblant a la colistina natural. Observem diferències entre l’activitat dels anàlegs segons la mida de la cadena d’àcid gras. Així mateix, la substitució de la cadena d’àcid gra per altres cadenes hidrofòbiques potencia l'activitat antimicrobiana dels pèptids resultants, permetent a més una síntesi més senzilla i eficient. El pèptid estrella de la tesi mostra una capacitat antimicrobiana contra bacteris gramnegatius similar a la colistina. A més, mostra uns perfils farmacocinètics i nefrotòxics potencialment compatibles amb la potencial administració. Atès que l'efecte del pèptid estrella en el tractament d'un model de pneumònia bacteriana no és inferior a la colistina, considerem que és un compost prometedor el qual s'hauria de continuar investigant i desenvolupant per tal que algun dia pugui arribar al mercat.

14 pages, 6 figures, 1 table, supporting information https://pubs.acs.org/doi/10.1021/acs.jcim.2c01344.-- The code for this work is available at https://github.com/bioexcel/biobb_hpc_workflows/tree/condapack. MD simulations are available from our MDposit repository: https://mdposit-dev.bsc.es/#/browse?search=egfr, Mutations in the kinase domain of the epidermal growth factor receptor (EGFR) can be drivers of cancer and also trigger drug resistance in patients receiving chemotherapy treatment based on kinase inhibitors. A priori knowledge of the impact of EGFR variants on drug sensitivity would help to optimize chemotherapy and design new drugs that are effective against resistant variants before they emerge in clinical trials. To this end, we explored a variety of in silico methods, from sequence-based to “state-of-the-art” atomistic simulations. We did not find any sequence signal that can provide clues on when a drug-related mutation appears or the impact of such mutations on drug activity. Low-level simulation methods provide limited qualitative information on regions where mutations are likely to cause alterations in drug activity, and they can predict around 70% of the impact of mutations on drug efficiency. High-level simulations based on nonequilibrium alchemical free energy calculations show predictive power. The integration of these “state-of-the-art” methods into a workflow implementing an interface for parallel distribution of the calculations allows its automatic and high-throughput use, even for researchers with moderate experience in molecular simulations, This work has been supported by the BioExcel-2: Centre of Excellence for Computational Biomolecular Research (823830), the Spanish Ministry of Science (RTI2018-096704-B-100 and PID2020-116620GB-I00), and the Instituto de Salud Carlos III–Instituto Nacional de Bioinformática (ISCIII PT 17/0009/0007 cofunded by the Fondo Europeo de Desarrollo Regional). Funding was also provided by the MINECO Severo Ochoa Award of Excellence from the Government of Spain (awarded to IRB Barcelona). M.O. is an ICREA (Institució Catalana de Recerca i Estudis Avancats) Academia researcher. Nostrum Biodiscovery is supported by the Fundación Marcelino Botín (Mind the Gap), CDTI (Neotec grant EXP 00094141/SNEO-20161127), and a Torres Quevedo grant (PTQ2018-009992), With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)

Urinary tract infections caused by extended-spectrum β-lactamase Escherichia coli (ESBL-EC) are increasing worldwide and are a current concern because treatment options are often limited. This study investigated antimicrobial susceptibility, antimicrobial resistance genes (ARGs), and the biological diversity of urinary ESBL-EC isolates at Cerdanya Hospital, a European cross-border hospital that combines French and Spanish healthcare models. Bacterial identification and susceptibility were determined using the Microscan WalkAway® system and ESBL production was examined by the double-disk synergy method. Isolates were sequenced using the Ion S5™ next-generation sequencing system, with the whole-genome sequences then assembled using SPADEs software and analyzed using PubMLST, ResFinder, FimTyper, PlasmidFinder, and VirulenceFinder. A phylogenetic analysis was performed by constructing an assembly-based core-SNV alignment, followed by a phylogenetic tree constructed using Parsnp from the Harvest suite. All isolates studied were multidrug-resistant and could be classified into 19 different sequence types characterized by a high genetic diversity. The most prevalent ESBL-enzymes were CTX-M-14 and CTX-M-15. High-risk international clones (ST131, ST10, and ST405) were also identified. The results demonstrated the absence of a single predominant clone of ESBL-MDR-EC at Cerdanya Hospital.

[Background]: Gasdermin B (GSDMB) over-expression promotes poor prognosis and aggressive behavior in HER2 breast cancer by increasing resistance to therapy. Decoding the molecular mechanism of GSDMB-mediated drug resistance is crucial to identify novel effective targeted treatments for HER2/GSDMB aggressive tumors. [Methods]: Different in vitro approaches (immunoblot, qRT-PCR, flow cytometry, proteomic analysis, immunoprecipitation, and confocal/electron microscopy) were performed in HER2 breast and gastroesophageal carcinoma cell models. Results were then validated using in vivo preclinical animal models and analyzing human breast and gastric cancer samples. [Results]: GSDMB up-regulation renders HER2 cancer cells more resistant to anti-HER2 agents by promoting protective autophagy. Accordingly, the combination of lapatinib with the autophagy inhibitor chloroquine increases the therapeutic response of GSDMB-positive cancers in vitro and in zebrafish and mice tumor xenograft in vivo models. Mechanistically, GSDMB N-terminal domain interacts with the key components of the autophagy machinery LC3B and Rab7, facilitating the Rab7 activation during pro-survival autophagy in response to anti-HER2 therapies. Finally, we validated these results in clinical samples where GSDMB/Rab7/LC3B co-expression associates significantly with relapse in HER2 breast and gastric cancers. [Conclusion]: Our findings uncover for the first time a functional link between GSDMB over-expression and protective autophagy in response to HER2-targeted therapies. GSDMB behaves like an autophagy adaptor and plays a pivotal role in modulating autophagosome maturation through Rab7 activation. Finally, our results provide a new and accessible therapeutic approach for HER2/GSDMB + cancers with adverse clinical outcome., This study has been supported by the Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación (PID2019-104644RB-I00) -GMB-, the Instituto de Salud Carlos III (CIBERONC, CB16/12/00449 -JA-, CB16/12/00231 -DLN- and CB16/12/00295 -GMB-, PI19/01181 -JA-, PI18/00795, CP17/00063 and RTI2018-095611-A-I00 -DLN- and ERA-NET TRANSCAN-2 -JA- [all partly supported by FEDER funds]) and by the AECC Scientific Foundation (FC_AECC PROYE19036MOR -GMB- and LABAE19004LLOB -DLN-). Furthermore, this work was supported by Breast Cancer Research Foundation (BCRF-19–08) -JA-. We are also grateful to the CERCA Programme (Generalitat de Catalunya) for institutional support. MGC and DS contracts are funded by CIBERONC, KG is a recipient of a PFIS fellowship (FI19/00188), RRB is recipient of a Ramón y Cajal grant (RyC-2016–19671) and DLN is recipient of a Miguel Servet grant (MS17/00063) (all partly supported by FEDER funds). We are also grateful to MD Anderson BIOBANK for providing tumor samples. The bank (reference # B.0000745) belongs to the National Registry of Biobanks coordinated by the Carlos III Health Institute.

BRCA1 mutant carcinomas are sensitive to PARP inhibitor (PARPi) therapy; however, resistance arises. BRCA1 BRCT domain mutant proteins do not fold correctly and are subject to proteasomal degradation, resulting in PARPi sensitivity. In this study, we show that cell lines and patient-derived tumors, with highly disruptive BRCT domain mutations, have readily detectable BRCA1 protein expression, and are able to proliferate in the presence of PARPi. Peptide analyses reveal that chemo-resistant cancers contain residues encoded by BRCA1 intron 15. Mechanistically, cancers with BRCT domain mutations harbor BRCA1 gene breakpoints within or adjacent to Alu elements in intron 15; producing partial gene duplications, inversions and translocations, and terminating transcription prior to the mutation-containing BRCT domain. BRCA1 BRCT domain-deficient protein isoforms avoid mutation-induced proteasomal degradation, support homology-dependent DNA repair, and promote PARPi resistance. Taken together, Alu-mediated BRCA1 gene rearrangements are responsible for generating hypomorphic proteins, and may represent a biomarker of PARPi resistance., BRCA1 mutations located within the BRCT domain result in proteasomal degradation and sensitivity to PARP inhibitors (PARPi). Here, the authors report genetic rearrangements in the BRCA1 gene that generate a BRCT-less BRCA1 protein isoform, which avoids degradation and leads to PARPi resistance.

Background Vancomycin-resistant enterococci (VRE) are major therapeutic challenges. Prospective contemporary data characterizing the clinical and molecular epidemiology of VRE bloodstream infections (BSIs) are lacking. Methods The Vancomycin-Resistant Enterococcal BSI Outcomes Study (VENOUS I) is a prospective observational cohort of adult patients with enterococcal BSI in 11 US hospitals. We included patients with Enterococcus faecalis or Enterococcus faecium BSI with ≥1 follow-up blood culture(s) within 7 days and availability of isolate(s) for further characterization. The primary study outcome was in-hospital mortality. Secondary outcomes were mortality at days 4, 7, 10, 12, and 15 after index blood culture. A desirability of outcome ranking was constructed to assess the association of vancomycin resistance with outcomes. All index isolates were subjected to whole genome sequencing. Results Forty-two of 232 (18%) patients died in hospital and 39 (17%) exhibited microbiological failure (lack of clearance in the first 4 days). Neutropenia (hazard ratio [HR], 3.13), microbiological failure (HR, 2.4), VRE BSI (HR, 2.13), use of urinary catheter (HR, 1.85), and Pitt BSI score ≥2 (HR, 1.83) were significant predictors of in-hospital mortality. Microbiological failure was the strongest predictor of in-hospital mortality in patients with E faecium bacteremia (HR, 5.03). The impact of vancomycin resistance on mortality in our cohort changed throughout the course of hospitalization. Enterococcus faecalis sequence type 6 was a predominant multidrug-resistant lineage, whereas a heterogeneous genomic population of E faecium was identified. Conclusions Failure of early eradication of VRE from the bloodstream is a major factor associated with poor outcomes.

Acid ceramidase (AC); Glucosylceramide synthase (GCS); Shingolipids Ceramidasa ácida (AC); Glucosilceramida sintasa (GCS); Esfingolípidos Ceramidasa àcida (AC); Glucosilceramida sintasa (GCS); Esfingolípids Drug resistance continues to be one of the major challenges to cure cancer. As research in this field evolves, it has been proposed that numerous bioactive molecules might be involved in the resistance of cancer cells to certain chemotherapeutics. One well-known group of lipids that play a major role in drug resistance are the sphingolipids. Sphingolipids are essential components of the lipid raft domains of the plasma membrane and this structural function is important for apoptosis and/or cell proliferation. Dysregulation of sphingolipids, including ceramide, sphingomyelin or sphingosine 1-phosphate, has been linked to drug resistance in different types of cancer, including breast, melanoma or colon cancer. Sphingolipid metabolism is complex, involving several lipid catabolism with the participation of key enzymes such as glucosylceramide synthase (GCS) and sphingosine kinase 1 (SPHK1). With an overview of the latest available data on this topic and its implications in cancer therapy, this review focuses on the main enzymes implicated in sphingolipids metabolism and their intermediate metabolites involved in cancer drug resistance. We thank Teresa Moline and Rosa Somoza from the VHIR. This work was supported by grants from the Instituto de Salud Carlos III (ISCIII; PI20/00556 and CP03/00101 [ML]) and CIBERONC (ML). This work was also co-financed by the European Regional Fund (ERDF) and AECC (Spanish Association of Cancer Research) (Founding Ref. GC16173720CARR [ML]). YG-M, CM, and AS-G were supported by the VHIR, iP-FIS (ISCIII) and VHIR fellowships, respectively.

Antibiotic stewardship programs optimize the use of antimicrobials to prevent the development of resistance and improve patient outcomes. In this prospective interventional study, a multidisciplinary team led by surgeons implemented a program aimed at shortening the duration of antibiotic treatment &lt, 7 days. The impact of the intervention on antibiotic consumption adjusted to bed-days and discharges, and the isolation of multiresistant bacteria (MRB) was also studied. Furthermore, the surgeons were surveyed regarding their beliefs and feelings about the program. Out of 1409 patients, 40.7% received antibiotic therapy. Treatment continued for over 7 days in 21.5% of cases, and, as can be expected, source control was achieved in only 48.8% of these cases. The recommendations were followed in 90.2% of cases, the most frequent being to withdraw the treatment (55.6%). During the first 16 months of the intervention, a sharp decrease in the percentage of extended treatments, with R2 = 0.111 was observed. The program was very well accepted by surgeons, and achieved a decrease in both the consumption of carbapenems and in the number of MRB isolations. Multidisciplinary stewardship teams led by surgeons seem to be well received and able to better manage antibiotic prescription in surgery.

Campylobacter jejuni; Epidemiologia; Resistència antibiòtica Campylobacter jejuni; Epidemiología; Resistencia antibiótica Campylobacter jejuni; Epidemiology; Antibiotic resistance Aquest informe té com a objectiu analitzar les característiques epidemiològiques dels casos confirmats de Campylobacter jejuni i analitzar la sensibilitat antimicrobiana dels casos declarats a l’SNMC durant els anys 2016-2019.

Methicillin-resistant Staphylococcus aureus bloodstream infections (MRSA-BSI) are a significant cause of mortality. We analysed the evolution of the molecular and clinical epidemiology of MRSA-BSI (n = 784) in adult patients (Barcelona, 1990–2019). Isolates were tested for antimicrobial susceptibility and genotyped (PFGE), and a selection was sequenced (WGS) to characterise the pangenome and mechanisms underlying antimicrobial resistance. Increases in patient age (60 to 71 years), comorbidities (Charlson’s index > 2, 10% to 94%), community-onset healthcare-associated acquisition (9% to 60%), and 30-day mortality (28% to 36%) were observed during the 1990–1995 and 2014–2019 periods. The proportion of catheter-related BSIs fell from 57% to 20%. Current MRSA-BSIs are caused by CC5-IV and an upward trend of CC8-IV and CC22-IV clones. CC5 and CC8 had the lowest core genome proportions. Antimicrobial resistance rates fell, and only ciprofloxacin, tobramycin, and erythromycin remained high (>50%) due to GyrA/GrlA changes, the presence of aminoglycoside-modifying enzymes (AAC(6′)-Ie-APH(2″)-Ia and ANT(4′)-Ia), and mph(C)/msr(A) or erm (C) genes. Two CC22-IV strains showed daptomycin resistance (MprF substitutions). MRSA-BSI has become healthcare-associated, affecting elderly patients with comorbidities and causing high mortality rates. Clonal replacement with CC5-IV and CC8-IV clones resulted in lower antimicrobial resistance rates. The increased frequency of the successful CC22-IV, associated with daptomycin resistance, should be monitored.

Tuberculosis (TB), is one of the top 10 causes of death worldwide (WHO). According to the last Global TB report from the World Health Organization, 10 million persons were estimated to have had TB in 2019 worldwide, causing about 1.6 million deaths. Tuberculosis has not only a dramatic impact on the quality of life for the patients, but also has raised many socio-economic issues at a community level, especially in medium and high burden regions, such as India, China, and Indonesia. In 2014, WHO adopted the "End TB strategy" which aimed to reduce TB deaths by 90% between 2015 and 2030, to prevent new cases by 80% during the same period and to decrease the socioeconomic impact of the disease at a family level. Even though tuberculosis global incidence has decreased significantly, efforts still need to be made to reach these goals. Non-tuberculous mycobacteria (NTM), in contrast to Mycobacterium tuberculosis, are bacteria widely spread in the environment and can be found in a broad range of ecosystems such as soils and water, including drinking water systems. NTM are opportunistic pathogens associated with both pulmonary and extrapulmonary infections. This Research Topic collected articles addressing: (i) TB and NTMs associated diseases, diagnostic, control, and public health, (ii) mycobacterial genomics, (iii) and antimycobacterial drugs and resistance

Neisseria gonorrhoeae; Epidemiologia; Resistència antibiòtica Neisseria gonorrhoeae; Epidemiología; Resistencia antibiótica Neisseria gonorrhoeae; Epidemiology; Antibiotic resistance Aquest informe té com a objectiu analitzar les característiques epidemiològiques dels casos confirmats de Neisseria gonorrhoeae i l’anàlisi de la sensibilitat antimicrobiana dels casos declarats a l’SNMC durant els anys 2016 i 2017.

Simple Summary Drug resistance is the principal limiting factor to achieving good survival rates in patients with cancer. The identification of potential biomarkers for diagnosis and prognostic prediction, as well as the design of new molecular-targeted treatments, will be essential to improving head and neck squamous cell carcinoma (HNSCC) patient outcomes. In this sense, the sensitization of resistant cells and cancer stem cells (CSCs) represents a major challenge in cancer therapy. We conducted a proteomic study involving cisplatin-resistance and CSCs with the aim to unravel the molecular and cellular mechanisms by which tumor cells acquire resistance to chemotherapy. Syntenin-1 (SDCBP) was identified as an important protein involved in the chemoresistance and stemness of HNSCC tumors. Abstract To characterize the mechanisms that govern chemoresistance, we performed a comparative proteomic study analyzing head and neck squamous cell carcinoma (HNSCC) cells: CCL-138 (parental), CCL-138-R (cisplatin-resistant), and cancer stem cells (CSCs). Syntenin-1 (SDCBP) was upregulated in CCL-138-R cells and CSCs over parental cells. SDCBP depletion sensitized biopsy-derived and established HNSCC cell lines to cisplatin (CDDP) and reduced CSC markers, Src activation being the main SDCBP downstream target. In mice, SDCBP-depleted cells formed tumors with decreased mitosis, Ki-67 positivity, and metastasis over controls. Moreover, the fusocellular pattern of CCL-138-R cell-derived tumors reverted to a more epithelial morphology upon SDCBP silencing. Importantly, SDCBP expression was associated with Src activation, poor differentiated tumor grade, advanced tumor stage, and shorter survival rates in a series of 382 HNSCC patients. Our results reveal that SDCBP might be a promising therapeutic target for effectively eliminating CSCs and CDDP resistance.