Workshop on cardiovascular extracellular matrix in health and disease in Baeza, Spain

The Workshop on Cardiovascular Extracellular Matrix in Health and Disease, International University of Andalusia, Baeza, Spain, 6-8 October 2014 served to discuss the current knowledge on the mechanisms integral to extracellular matrix homeostasis that are fundamental to understanding the pathological basis of several cardiovascular diseases, including the development of cardiac fibrosis in response to cardiac hypertrophy and myocardial infarction, and the extracellular matrix alterations contributing to aortic stenosis or aneurysms.We highly appreciate the contribution of the International University of Andalusia (UNIA), the European Research Council (ERC) and the consortium FIBROTEAM (Comunidad Autónoma de Madrid, 2010-BMD2321)

Matrix stiffening and β1 integrin drive subtype-specific fibroblast accumulation in lung cancer

©2014 AACR. The crucial role of tumor-associated fibroblasts (TAF) in cancer progression is now clear in non-small cell lung cancer (NSCLC). However, therapies against TAFs are limited due to a lack of understanding in the subtype-specific mechanisms underlying their accumulation. Here, the mechanical (i.e., matrix rigidity) and soluble mitogenic cues that drive the accumulation of TAFs from major NSCLC subtypes: adenocarcinoma (ADC) and squamous cell carcinoma (SCC) were dissected. Fibroblasts were cultured on substrata engineered to exhibit normal- or tumor-like stiffnesses at different serumconcentrations, and critical regulatory processes were elucidated. In control fibroblasts from nonmalignant tissue, matrix stiffening alone increased fibroblast accumulation, and this mechanical effect was dominant or comparable with that of soluble growth factors up to 0.5% serum. The stimulatory cues ofmatrix rigidity were driven by β1 integrin mechanosensing through FAK (pY397), and were associated with a posttranscriptionally driven rise in β1 integrin expression. The latter mechano-regulatory circuit was also observed in TAFs but in a subtype-specific fashion, because SCC-TAFs exhibited higher FAK (pY397), β1 expression, and ERK1/2 (pT202/Y204) than ADC-TAFs. Moreover, matrix stiffening induced a larger TAF accumulation in SCC-TAFs (>50%) compared with ADC-TAFs (10%-20%). In contrast, SCC-TAFs were largely serum desensitized, whereas ADC-TAFs responded to high serum concentration only. These findings provide the first evidence of subtype-specific regulation of NSCLC-TAF accumulation. Furthermore, these data support that therapies aiming to restore normal lung elasticity and/or β1 integrin-dependent mechano regulation may be effective against SCC-TAFs, whereas inhibiting stromal growth factor signaling may be effective against ADC-TAFs. Implications: This study reveals distinct mechanisms underlying the abnormal accumulation of tumor-supporting fibroblasts in two major subtypes of lung cancer, which will assist the development of personalized therapies against these cells.Ministerio de Economía y Competitividad (SAF2009-13243 and PI13/02368, to J. Alcaraz; PS09/01377, to N. Reguart; SAF2012-34916, to F. Rodríguez-Pascual), AECC (10/103; to N. Reguart and J. Alcaraz), SEPAR (2009-853; to N. Reguart), and predoctoral fellowships from the Fundacio Cellex (to M. Puig),Ministerio de Educación (to A. Giménez), CONACYT (to R. Lugo), and COLCIENCIAS (to A. Velasquez).Peer Reviewe

Valoración médico-legal de las lesiones tras descarga eléctrica en el ámbito laboral: a propósito de un caso

como otros órganos, la audición puede ser afectada por el paso de corriente eléctrica a través del organismo. Son muchas las referencias bibliográficas encontradas de afectación de diversos órganos tras descarga eléctrica pero muy pocas del órgano de la audición. En el presente artículo se realiza la valoración del daño corporal de las lesiones tras descarga eléctrica en ámbito laboral a propósito de un caso, con especial atención a la valoración de la función auditiva, raramente referenciada de forma específica. Se expone la existencia de la afectación de la función auditiva, evolución y estabilidad de la misma tras la descarga eléctrica y su valoración médico-legal. Es necesario disponer de pruebas complementarias rigurosas y regladas para la valoración médico-legal posterior. Y la necesidad de seguimiento en el tiempo por la reversibilidad de dicha función, a pesar de mostrarse clínicamente como una hipoacusia neurosensorial.like other organs, hearing can be affected by the passage of an electrical current through the organism. There are many bibliographical references found affecting various organs after electrical shock but very few of the auditory organ. In the present article an evaluation is carried out on the medical-legal assessment of injuries after an electrical shock at work is made case report, with special attention on the evaluation of the auditory function, rarely specifically referenced. The existence of the effect of the auditive function, as well as its progress and further stability after the electrical shock and a medical-legal evaluation. It is necessary to have rigorous and regulated complementary tests for the subsequent medical-legal assessment. And the need for follow-up over time for the reversibility of this function, despite being clinically shown as a sensorineural hearing loss

Pharmacodynamics of fosfomycin: Insights into clinical use for antimicrobial resistance

The aim of this study was to improve the understanding of the pharmacokinetic-pharmacodynamic relationships of fosfomycin against extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains that have different fosfomycin MICs. Our methods included the use of a hollow fiber infection model with three clinical ESBL-producing E. coli strains. Human fosfomycin pharmacokinetic profiles were simulated over 4 days. Preliminary studies conducted to determine the dose ranges, including the dose ranges that suppressed the development of drug-resistant mutants, were conducted with regimens from 12 g/day to 36 g/day. The combination of fosfomycin at 4 g every 8 h (q8h) and meropenem at 1 g/q8h was selected for further assessment. The total bacterial population and the resistant subpopulations were determined. No efficacy was observed against the Ec42444 strain (fosfomycin MIC, 64 mg/liter) at doses of 12, 24, or 36 g/day. All dosages induced at least initial bacterial killing against Ec46 (fosfomycin MIC, 1 mg/liter). High-level drug-resistant mutants appeared in this strain in response to 12, 15, and 18 g/day. In the study arms that included 24 g/day, once or in a divided dose, a complete extinction of the bacterial inoculum was observed. The combination of meropenem with fosfomycin was synergistic for bacterial killing and also suppressed all fosfomycinresistant clones of Ec2974 (fosfomycin MIC, 1 mg/liter). We conclude that fosfomycin susceptibility breakpoints (≤64 mg/liter according to CLSI [for E. coli urinary tract infections only]) should be revised for the treatment of serious systemic infections. Fosfomycin can be used to treat infections caused by organisms that demonstrate lower MICs and lower bacterial densities, although relatively high daily dosages (i.e., 24 g/day) are required to prevent the emergence of bacterial resistance. The ratio of the area under the concentration-time curve for the free, unbound fraction of fosfomycin versus the MIC (fAUC/MIC) appears to be the dynamically linked index of suppression of bacterial resistance. Fosfomycin with meropenem can act synergistically against E. coli strains in preventing the emergence of fosfomycin resistance.Consejería de Igualdad, Salud y Políticas Sociales Junta de Andalucía PI-0044-2013FEDER REIPI RD12/001

Elevated expression levels of lysyl oxidases protect against aortic aneurysm progression in Marfan syndrome

© 2015 Elsevier Ltd. Patients with Marfan syndrome (MFS) are at high risk of life-threatening aortic dissections. The condition is caused by mutations in the gene encoding fibrillin-1, an essential component in the formation of elastic fibers. While experimental findings in animal models of the disease have shown the involvement of transforming growth factor-β (TGF-β)- and angiotensin II-dependent pathways, alterations in the vascular extracellular matrix (ECM) may also play a role in the onset and progression of the aortic disease. Lysyl oxidases (LOX) are extracellular enzymes, which initiates the formation of covalent cross-linking of collagens and elastin, thereby contributing to the maturation of the ECM. Here we have explored the role of LOX in the formation of aortic aneurysms in MFS. We show that aortic tissue from MFS patients and MFS mouse model (Fbn1C1039G/+) displayed enhanced expression of the members of the LOX family, LOX and LOX-like 1 (LOXL1), and this is associated with the formation of mature collagen fibers. Administration of a LOX inhibitor for 8weeks blocked collagen accumulation and aggravated elastic fiber impairment, and these effects correlated with the induction of a strong and rapidly progressing aortic dilatation, and with premature death in the more severe MFS mouse model, Fbn1mgR/mgR, without any significant effect on wild type animals. This detrimental effect occurred preferentially in the ascending portion of the aorta, with little or no involvement of the aortic root, and was associated to an overactivation of both canonical and non-canonical TGF-β signaling pathways. The blockade of angiotensin II type I receptor with losartan restored TGF-β signaling activation, normalized elastic fiber impairment and prevented the aortic dilatation induced by LOX inhibition in Fbn1C1039G/+ mice. Our data indicate that LOX enzymes and LOX-mediated collagen accumulation play a critical protective role in aneurysm formation in MFS.Ministerio de Economía y Competitividad (Plan Nacional de I + D + I: SAF2012-34916), and Comunidad Autónoma deMadrid: 2010-BMD2321, FIBROTEAMConsortium, to F.R-P.; Howard Hughes Medical Institute, Smilow Center for Marfan Syndrome Research, The Marfan Foundation, and the National Institutes of Health (AR41135 and AR049698) to H.C.D.; and Ministerio de Economía y Competitividad (Plan Nacional de I + D + I: BFU2012- 33932), Fundación Ramón Areces and the National Marfan FoundationPeer Reviewe

LoXL4 is induced by transforming growth factor β1 through Smad and JunB/Fra2 and contributes to vascular matrix remodeling

Transforming growth factor β1 (TGF-β1) is a pleiotropic factor involved in the regulation of extracellular matrix (ECM) synthesis and remodeling. In search for novel genes mediating the action of TGF-β1 on vascular ECM, we identified the member of the lysyl oxidase family of matrix-remodeling enzymes, lysyl oxidase-like 4 (LOXL4), as a direct target of TGF-β1 in aortic endothelial cells, and we dissected the molecular mechanism of its induction. Deletion mapping and mutagenesis analysis of the LOXL4 promoter demonstrated the absolute requirement of a distal enhancer containing an activator protein 1 (AP-1) site and a Smad binding element for TGF-β1 to induce LOXL4 expression. Functional cooperation between Smad proteins and the AP-1 complex composed of JunB/Fra2 accounted for the action of TGF-β1, which involved the extracellular signal-regulated kinase (ERK)- dependent phosphorylation of Fra2. We furthermore provide evidence that LOXL4 was extracellularly secreted and significantly contributed to ECM deposition and assembly. These results suggest that TGF-β1-dependent expression of LOXL4 plays a role in vascular ECM homeostasis, contributing to vascular processes associated with ECM remodeling and fibrosis.This work was supported by grants from the Ministerio de Economía y Competitividad (Plan Nacional de I+D+I: SAF2009-09085, SAF2012-34916), Comunidad Autónoma de Madrid (2010-BMD2321, FIBROTEAM Consortium), Fundación Genoma España (MEICA project), Consejo Superior de Investigaciones Científicas (Proyecto Intramural de Incorporación, 200920I158), and Fundación Renal Iñigo Alvárez de Toledo. O.B. is a recipient of a fellowship from the Ministerio de Economía y Competi- tividad (Formación de Personal Investigador)Peer Reviewe

Muscle Damage Biomarkers in Congestion Weeks in English Premier League Soccer Players: A Prospective Study for Two Consecutive Seasons

The current study was conducted to compare muscle damage biomarkers in single- vs. multi-match weeks in elite soccer players for two consecutive seasons. A secondary objective was to analyze the influence of playing position and exposure time on muscle damage in single- vs. multi-match weeks. This is a prospective cohort study performed in a professional elite soccer club in the English Premier League during the 2018-2019 and 2019-2020 seasons up until the lockdown due to the COVID-19 pandemic. Data were collected in the Medical Department Room of an English Premier League Club before and after the soccer game from a total of 29 elite soccer players (mean +/- S.D.; age = 27.59 +/- 3.83 years; height = 1.83 +/- 0.05 m; body mass = 80.16 +/- 7.45 kg) who were enrolled in the club during both seasons. The main outcome measurements were creatine kinase (CK), weight, lean mass, % fat DEXA, high speed running, total distance, density of total distance and high-speed running and wellbeing questionnaires. Significance was set at p < 0.05. Players who completed more than 60 min in the previous game had significantly increased pregame CK levels and fatigue in multi-match weeks. Midfielders had both significantly increased pregame CK and muscle soreness in multi-match weeks. Midfielders and players with an exposure time of at least 60 min showed higher pregame CK values that should play a key role for deciding substitutions.We thank all the Medical Department at Watford FC, specially Chris Mogekwu and all the Physiotherapy Department at Camilo Jose Cela University. Both helped us get the sample and with the kindly support for this research. The results of the study are presented clearly, honestly, and without fabrication, falsification, or inappropriate data manipulation, and statement that results of the present study do not constitute endorsement by IJERPH. Documen

Interplay Among Different Fosfomycin Resistance Mechanisms in Klebsiella Pneumoniae

The objectives of this study were to characterize the role of the uhpT, glpT, and fosA genes in fosfomycin resistance in Klebsiella pneumoniae and evaluate the use of sodium phosphonoformate (PPF) in combination with fosfomycin. Seven clinical isolates of K. pneumoniae and the reference strain (ATCC 700721) were used, and their genomes were sequenced. DuhpT, DglpT, and DfosA mutants were constructed from two isolates and K. pneumoniae ATCC 700721. Fosfomycin susceptibility testing was done by the gradient strip method. Synergy between fosfomycin and PPF was studied by checkerboard assay and analyzed using SynergyFinder. Spontaneous fosfomycin mutant frequencies at 64 and 512mg/liter, in vitro activity using growth curves with fosfomycin gradient concentrations (0 to 256mg/liter), and time-kill assays at 64 and 307mg/liter were evaluated with and without PPF (0.623mM). The MICs of fosfomycin against the clinical isolates ranged from 16 to ≥1,024mg/liter. The addition of 0.623mM PPF reduced fosfomycin MIC between 2- and 8-fold. Deletion of fosA led to a 32-fold decrease. Synergistic activities were observed with the combination of fosfomycin and PPF (most synergistic area at 0.623mM). The lowest fosfomycin-resistant mutant frequencies were found in ΔfosA mutants, with decreases in frequency from 1.69×10-1 to 1.60×10-5 for 64mg/liter of fosfomycin. In the final growth monitoring and time-kill assays, fosfomycin showed a bactericidal effect only with the deletion of fosA and not with the addition of PPF. We conclude that fosA gene inactivation leads to a decrease in fosfomycin resistance in K. pneumoniae. The pharmacological approach using PPF did not achieve enough activity, and the effect decreased with the presence of fosfomycin-resistant mutations.Ministerio de Economía y Competitividad PI16/01824, REIPI RD12/0015/0010, EIPI RD16/0016/0001Junta de Andalucía PI-0044Innovative Medicines Initiative 115523, 115620, 11573

Epistasis between intracellular cholesterol trafficking-related genes (NPC1 and ABCA1) and Alzheimer's disease risk

Aberrant cholesterol metabolism has been implicated in Alzheimer´s disease (AD). Recent findings have suggested an interaction of Niemann-Pick C1 (NPC1) and ATP-binding cassette transporter A1 (ABCA1) proteins in intracellular cholesterol transport and in maintaining cell cholesterol balance. Underexpression of NPC1 in concert with underexpression of ABCA1 would result in increased cholesterol accumulation and increased AD risk. We examined a functional polymorphism in the ABCA1 promoter region (−477, rs2422493), and four NPC1 polymorphisms in exon 6 (rs18050810), intron 20 (rs4800488), intron 22 (rs2236707), and intron 24 (rs2510344) capturing 85% of genetic variability in Hap Map CEU population, in a group of 631 Spanish AD patients and 731 controls. Subjects carrying both the ABCA1 (−477) TT genotype and the NPC1 (exon 6) GG genotype (OR = 1.89; 95% CI 1.04-3.41), NPC1 (intron 20) AA genotype (OR = 2.05; 95% CI 1.26-3.33), NPC1 (intron 22) AA genotype (OR = 2.05; 95% CI 1.18-3.58), or NPC1 (intron 24) GG genotype (OR = 1.89; 95% CI 1.16-3.07) had a higher risk of developing AD than subjects without these risk genotypes. Testing for epistatic interaction between genes in the pathway of cholesterol metabolism might be useful for predicting AD risk