Streptococcus pneumoniae y Staphylococcus aureus : enfermedad invasiva, biofilm y terapia

Streptococcus pneumoniae (neumococo) es el principal responsable de neumonía bacteriana adquirida en comunidad. Su principal factor de virulencia es el polisacárido capsular (CPS), dividiéndolo en, al menos, 101 serotipos. Entre un 5–10% de adultos y un 20–40% de niños, están colonizados por este patógeno, siendo este estado de portador en la nasofaringe, un prerrequisito para la posterior diseminación a vías estériles y producir enfermedad neumocócica invasiva (ENI). La ENI es prevenible por vacunación, en la actualidad se usan en España dos vacunas: la vacuna conjugada13-valente (VCN13) y la vacuna polisacarídica 23-valente (VPN23). Desde la introducción de las vacunas conjugadas se ha observado un fenómeno denominado reemplazo de serotipos. Staphylococcus aureus, es otro patógeno relevante responsable de un rango amplio de infecciones, desde menos graves como la foliculitis hasta infecciones invasivas graves como la endocarditis. S. aureus posee un gran arsenal de factores de virulencia como el CPS, presente en más del 90% de los aislados clínicos. Su principal relevancia clínica reside en su resistencia a antibióticos, especialmente S. aureus resistente a meticilina (SARM). S. aureus coloniza hasta el 80% de individuos..

Experimental study of concentration polarization in a crossflow reverse osmosis system using Digital Holographic Interferometry

Digital Holographic Interferometry (DHI) has been used to visualize the polarization concentration layer during crossflow RO. This technique is based on the fact that changes in the concentration of the solution produce changes in its refractive index. Therefore, the concentration profile formed due to the polarization phenomenon can be visualized as interference fringes. Experiments with Na2SO4 and NaCl solutions have been carried out. Three variables of the process were studied: crossflow velocity, initial concentration and pressure applied. In each experiment, crossflow velocity was changed every 30min, in an increasing or decreasing sequence. Few minutes after changing the crossflow velocity the steady-state was reached. Interference fringe patterns of the polarization layer and their corresponding concentration profiles, as well as the permeate flux in different experimental conditions, are presented. The major experimental result is the visualization for the first time in situ and in real-time of the polarization layer in a process of crossflow by a non-invasive method. Experimental results show a close relationship among crossflow velocity, permeate flux and polarization layer. Furthermore, experimental maximum concentration values reached at the membrane surface were compared with values calculated by using the film theory approach and a good agreement was obtained.This research has been sponsored by the Plan Nacional de I+D+I CTQ2006-14904 (Ministerio de Ciencia e Innovación) and Generalitat Valenciana, Consellería de Educación (ACOMP/2009/366)

Combination of Antibodies and Antibiotics as a Promising Strategy Against Multidrug-Resistant Pathogens of the Respiratory Tract

The emergence of clinical isolates associated to multidrug resistance is a serious threat worldwide in terms of public health since complicates the success of the antibiotic treatment and the resolution of the infectious process. This is of great concern in pathogens affecting the lower respiratory tract as these infections are one of the major causes of mortality in children and adults. In most cases where the respiratory pathogen is associated to multidrug-resistance, antimicrobial concentrations both in serum and at the site of infection may be insufficient and the resolution of the infection depends on the interaction of the invading pathogen with the host immune response. The outcome of these infections largely depends on the susceptibility of the pathogen to the antibiotic treatment, although the humoral and cellular immune responses also play an important role in this process. Hence, prophylactic measures or even immunotherapy are alternatives against these multi-resistant pathogens. In this sense, specific antibodies and antibiotics may act concomitantly against the respiratory pathogen. Alteration of cell surface structures by antimicrobial drugs even at sub-inhibitory concentrations might result in greater exposure of microbial ligands that are normally hidden or hardly exposed. This alteration of the bacterial envelope may stimulate opsonization by natural and/or specific antibodies or even by host defense components, increasing the recognition of the microbial pathogen by circulating phagocytes. In this review we will explain the most relevant studies, where vaccination or the use of monoclonal antibodies in combination with antimicrobial treatment has demonstrated to be an alternative strategy to overcome the impact of multidrug resistance in respiratory pathogens

Clinical Relevance and Molecular Pathogenesis of the Emerging Serotypes 22F and 33F of Streptococcus pneumoniae in Spain

Streptococcus pneumoniae is the main bacterial cause of respiratory infections in children and the elderly worldwide. Serotype replacement is a frequent phenomenon after the introduction of conjugated vaccines, with emerging serotypes 22F and 33F as frequent non-PCV13 serotypes in children and adults in North America and other countries. Characterization of mechanisms involved in evasion of the host immune response by these serotypes is of great importance in public health because they are included in the future conjugated vaccines PCV15 and PCV20. One of the main strategies of S. pneumoniae to persistently colonize and causes infection is biofilm formation. In this study, we have evaluated the influence of capsule polysaccharide in biofilm formation and immune evasion by using clinical isolates from different sources and isogenic strains with capsules from prevalent serotypes. Since the introduction of PCV13 in Spain in the year 2010, isolates of serotypes 22F and 33F are rising among risk populations. The predominant circulating genotypes are ST43322F and ST71733F , being CC433 in 22F and CC717 in 33F the main clonal complexes in Spain. The use of clinical isolates of different origin, demonstrated that pediatric isolates of serotypes 22F and 33F formed better biofilms than adult isolates and this was statistically significant. This phenotype was greater in clinical isolates from blood origin compared to those from cerebrospinal fluid, pleural fluid and otitis. Opsonophagocytosis assays showed that serotype 22F and 33F were recognized by the PSGL-1 receptor on leukocytes, although serotype 22F, was more resistant than serotype 33F to phagocytosis killing and more lethal in a mouse sepsis model. Overall, the emergence of additional PCV15 serotypes, especially 22F, could be associated to an enhanced ability to divert the host immune response that markedly increased in a biofilm state. Our findings demonstrate that pediatric isolates of 22F and 33F, that form better biofilm than isolates from adults, could have an advantage to colonize the nasopharynx of children and therefore, be important in carriage and subsequent dissemination to the elderly. The increased ability of serotype 22F to avoid the host immune response, might explain the emergence of this serotype in the last years.This work was partially supported by a grant from the MSD-USA (MISP Call, Reference: 57320), Ministerio de Economía, Industria y Competitividad (MINECO) (SAF2017-83388-R), and CIBER de Enfermedades Respiratorias (CIBERES) is an initiative of the Instituto de Salud Carlos III

Combination of Cefditoren and N-acetyl-l-Cysteine Shows a Synergistic Effect against Multidrug-Resistant Streptococcus pneumoniae Biofilms

Biofilm formation by Streptococcus pneumoniae is associated with colonization of the upper respiratory tract, including the carrier state, and with chronic respiratory infections in patients suffering from chronic obstructive pulmonary disease (COPD). The use of antibiotics alone to treat recalcitrant infections caused by biofilms is insufficient in many cases, requiring novel strategies based on a combination of antibiotics with other agents, including antibodies, enzybiotics, and antioxidants. In this work, we demonstrate that the third-generation oral cephalosporin cefditoren (CDN) and the antioxidant N-acetyl-l-cysteine (NAC) are synergistic against pneumococcal biofilms. Additionally, the combination of CDN and NAC resulted in the inhibition of bacterial growth (planktonic and biofilm cells) and destruction of the biofilm biomass. This marked antimicrobial effect was also observed in terms of viability in both inhibition (prevention) and disaggregation (treatment) assays. Moreover, the use of CDN and NAC reduced bacterial adhesion to human lung epithelial cells, confirming that this strategy of combining these two compounds is effective against resistant pneumococcal strains colonizing the lung epithelium. Finally, administration of CDN and NAC in mice suffering acute pneumococcal pneumonia caused by a multidrug-resistant strain was effective in clearing the bacteria from the respiratory tract in comparison to treatment with either compound alone. Overall, these results demonstrate that the combination of oral cephalosporins and antioxidants, such as CDN and NAC, respectively, is a promising strategy against respiratory biofilms caused by S. pneumoniae. IMPORTANCE Streptococcus pneumoniae is one of the deadliest bacterial pathogens, accounting for up to 2 million deaths annually prior to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Vaccines have decreased the burden of diseases produced by S. pneumoniae, but the rise of antibiotic-resistant strains and nonvaccine serotypes is worrisome. Pneumococcal biofilms are associated with chronic respiratory infections, and treatment is challenging, making the search for new antibiofilm therapies a priority as biofilms become resistant to traditional antibiotics. In this work, we used the combination of an antibiotic (CDN) and an antioxidant (NAC) to treat the pneumococcal biofilms of relevant clinical isolates. We demonstrated a synergy between CDN and NAC that inhibited and treated pneumococcal biofilms, impaired pneumococcal adherence to the lung epithelium, and treated pneumonia in a mouse pneumonia model. We propose the widely used cephalosporin CDN and the repurposed drug NAC as a new antibiofilm therapy against S. pneumoniae biofilms, including those formed by antibiotic-resistant clinical isolates.This work was supported by Ministerio de Ciencia e Innovación (MICINN) (grant PID2020-119298RB-I00) and by Meiji Pharma Spain (grant MVP 119/20). J.Y. has received grants from MSD-USA (MISP Call) and Pfizer that are not related to this work. J.Y. has participated in advisory boards organized by MSD and Pfizer. M.G. and P.C. are members of the Scientific Department of Meiji Pharma Spain. The other authors declare no competing interests.S

DeepP: Deep Learning Multi-Program Prefetch Configuration for the IBM POWER 8

[EN] Current multi-core processors implement sophisticated hardware prefetchers, that can be configured by application (PID),to improve the system performance. When running multiple applications, each application can present different prefetch requirements,hence different configurations can be used. Setting the optimal prefetch configuration for each application is a complex task since itdoes not only depend on the application characteristics but also on the interference at the shared memory resources (e.g. memorybandwidth). In his paper, we proposeDeepP, a deep learning approach for the IBM POWER8 that identifies at run-time the bestprefetch configuration for each application in a workload. To this end, the neural network predicts the performance of each applicationunder the studied prefetch configurations by using a set of performance events. The prediction accuracy of the network is improvedthanks to a dynamic training methodology that allows learning the impact of dynamic changes of the prefetch configuration onperformance. At run-time, the devised network infers the best prefetch configuration for each application and adjusts it dynamically.Experimental results show that the proposed approach improves performance, on average, by 5,8%, 6,7%, and 15,8% compared tothe default prefetch configuration across different 6-, 8-, and 10-application workloads, respectively.This work was supported in part by Ministerio de Ciencia, Innovacion y Universidades and the European ERDF under Grant RTI2018-098156-B-C51, in part by Generalitat Valenciana under Grant AICO/2021/266, and in part by Ayudas Contratos predoctorales UPV -subprograma 1 (PAID-01-20). The work of Josue Feliu was supported by a Juan de la Cierva Formacion Contract under Grant FJC2018-036021-I.Lurbe-Sempere, M.; Feliu-Pérez, J.; Petit Martí, SV.; Gómez Requena, ME.; Sahuquillo Borrás, J. (2022). DeepP: Deep Learning Multi-Program Prefetch Configuration for the IBM POWER 8. IEEE Transactions on Computers. 71(10):2646-2658. https://doi.org/10.1109/TC.2021.313999726462658711

Nationwide trends of invasive pneumococcal disease in Spain (2009-2019) in children and adults during the pneumococcal conjugate vaccine era.

Introduction of pneumococcal conjugate vaccines (PCVs) has shown a marked reduction in the disease caused by vaccine serotypes in children providing herd protection to the elderly group. However, the emergence of non-vaccine serotypes is of great concern worldwide. This study includes national laboratory data from invasive pneumococcal disease (IPD) cases affecting pediatric and adult population during 2009-2019. The impact of implementing different vaccine strategies for immunocompetent adults comparing Spanish regions using PCV13 vs regions using PPV23 vaccine was also analyzed for 2017-2019. The overall reductions of IPD cases by PCV13 serotypes in children and adults were 88% and 59% respectively during 2009-2019 with a constant increase of serotype 8 in adults since 2015. IPD cases by additional serotypes covered by PPV23 increased from 20% in 2009 to 52% in 2019. In children, serotype 24F was the most frequent in 2019 whereas in adults, serotypes 3 and 8 accounted for 36% of IPD cases. Introduction of PCV13 or PPV23 in the adult calendar of certain Spanish regions reduced up to 25% and 11% respectively the IPD cases by PCV13 serotypes, showing a decrease of serotype 3 when PCV13 was used. Use of PCV13 in children has shown a clear impact in pneumococcal epidemiology reducing the burden of IPD in children but also in adults by herd protection although the increase of serotype 8 in adults is worrisome. Vaccination with PCV13 in immunocompetent adults seems to control IPD cases by PCV13 serotypes including serotype 3.This work was supported by Ministerio de Economía, Industria y Competitividad (MINECO) [grant SAF2017-83388] and internal funding fromS

Effect of pneumococcal conjugate vaccines and SARS-CoV-2 on antimicrobial resistance and the emergence of Streptococcus pneumoniae serotypes with reduced susceptibility in Spain, 2004-20: a national surveillance study

Background: Epidemiological studies are necessary to explore the effect of current pneumococcal conjugate vaccines (PCVs) against antibiotic resistance, including the rise of non-vaccine serotypes that are resistant to antibiotics. Hence, epidemiological changes in the antimicrobial pattern of Streptococcus pneumoniae before and during the first year of the COVID-19 pandemic were studied. Methods: In this national surveillance study, we characterised the antimicrobial susceptibility to a panel of antibiotics in 3017 pneumococcal clinical isolates with reduced susceptibility to penicillin during 2004-20 in Spain. This study covered the early and late PCV7 periods; the early, middle, and late PCV13 periods; and the first year of the COVID-19 pandemic, to evaluate the contribution of PCVs and the pandemic to the emergence of non-vaccine serotypes associated with antibiotic resistance. Findings: Serotypes included in PCV7 and PCV13 showed a decline after the introduction of PCVs in Spain. However, an increase in non-PCV13 serotypes (mainly 11A, 24F, and 23B) that were not susceptible to penicillin promptly appeared. A rise in the proportion of pneumococcal strains with reduced susceptibility to β-lactams and erythromycin was observed in 2020, coinciding with the emergence of SARS-CoV-2. Cefditoren was the β-lactam with the lowest minimum inhibitory concentration (MIC)50 or MIC90 values, and had the highest proportion of susceptible strains throughout 2004-20. Interpretation: The increase in non-PCV13 serotypes associated with antibiotic resistance is concerning, especially the increase of penicillin resistance linked to serotypes 11A and 24F. The future use of PCVs with an increasingly broad spectrum (such as PCV20, which includes serotype 11A) could reduce the impact of antibiotic resistance for non-PCV13 serotypes. The use of antibiotics to prevent co-infections in patients with COVID-19 might have affected the increased proportion of pneumococcal-resistant strains. Cefotaxime as a parenteral option, and cefditoren as an oral choice, were the antibiotics with the highest activity against non-PCV20 serotypes.This work was supported by the Spanish Ministry of Science and Innovation (grant PID2020–119298RB-I00), Meiji Pharma Spain (grant MVP 119/20), and internal funding from Instituto de Salud Carlos III.S

Minilungs from Human Embryonic Stem Cells to Study the Interaction of Streptococcus pneumoniae with the Respiratory Tract

The new generation of organoids derived from human pluripotent stem cells holds a promising strategy for modeling host-bacteria interaction studies. Organoids recapitulate the composition, diversity of cell types, and, to some extent, the functional features of the native organ. We generated lung bud organoids derived from human embryonic stem cells to study the interaction of Streptococcus pneumoniae (pneumococcus) with the alveolar epithelium. Invasive pneumococcal disease is an important health problem that may occur as a result of the spread of pneumococcus from the lower respiratory tract to sterile sites. We show here an efficient experimental approach to model the main events of the pneumococcal infection that occur in the human lung, exploring bacterial adherence to the epithelium and internalization and triggering an innate response that includes the interaction with surfactant and the expression of representative cytokines and chemokines. Thus, this model, based on human minilungs, can be used to study pneumococcal virulence factors and the pathogenesis of different serotypes, and it will allow therapeutic interventions in a reliable human context. Importance: Streptococcus pneumoniae is responsible for high morbidity and mortalities rates worldwide, affecting mainly children and adults older than 65 years. Pneumococcus is also the most common etiologic agent of bacterial pneumonia and nonepidemic meningitis, and it is a frequent cause of bacterial sepsis. Although the introduction of pneumococcal vaccines has decreased the burden of pneumococcal disease, the rise of antibiotic-resistant strains and nonvaccine types by serotype replacement is worrisome. To study the biology of pneumococcus and to establish a reliable human model for pneumococcal pathogenesis, we generated human minilungs from embryonic stem cells. The results show that these organoids can be used to model some events occurring during the interaction of pneumococcus with the lung, such as adherence, internalization, and the initial alveolar innate response. This model also represents a great alternative for studying virulence factors involved in pneumonia, drug screening, and other therapeutic interventions.This work was supported by grant PI19CIII/00003 from the ISCIII to A.Z. and grantsPID2020-119298RB-I00 and PID2020-114546RB-I00 from the Spanish Ministry of Scienceand Innovation (MICINN) to J.Y. and O.Z., respectively. M.P.D.L. and J.M.R.-C. received grantsupport from AESI (PI20CIII/0029), the Spanish Association Against Cancer (AECC,CGB14143035THOM), and CIBERONC (group CB16/12/00273). S.A.R. was granted apostdoctoral fellowship by the Fundacão de Amparo á pesquisa do Estado do SãoPaulo (reference FAPESP-BEBE 2020/09919-0).S

Stable and Broad Spectrum Cross-Protection Against Pepino Mosaic Virus Attained by Mixed Infection

While recent pepino mosaic virus (PepMV; species Pepino mosaic virus, genus Potexvirus, family Alphaflexiviridae) epidemics seem to be predominantly caused by isolates of the CH2 strain, PepMV epidemics in intensive tomato crops in Spain are caused by both CH2 and EU isolates that co-circulate, representing a challenge in terms of control, including cross-protection. In this work, we hypothesized that mixed infections with two mild isolates of the EU and CH2 strains (PepMV-Sp13 and -PS5, respectively) may be useful in PepMV cross-protection in Spanish epidemics, providing protection against a broad range of aggressive isolates. Thus, we performed a range of field trials and an experimental evolution assay to determine the phenotypic and genetic stability of PepMV-Sp13 and -PS5 mixed infections, as well as their cross-protective efficiency. Our results showed that: (i) the phenotype of PepMV-Sp13 and -PS5 mixed infections was mild and did not change significantly when infecting different tomato cultivars or under different environmental conditions in Spain, (ii) PepMV-Sp13 and -PS5 mixed infections provided more efficient protection against two aggressive EU and CH2 isolates than single infections, and (iii) PepMV-Sp13 and -PS5, either in single or in mixed infections, were less variable than other two PepMV isolates occurring naturally in PepMV epidemics in Spain