Sex-related differences of fatty acid-binding protein 4 and leptin levels in atrial fibrillation.

AIMS: Adiposity plays a key role in the pathogenesis of atrial fibrillation (AF). Our aim was to study the sex differences in adipokines levels according to AF burden. METHODS AND RESULTS: Two independent cohorts of patients were studied: (i) consecutive patients with AF undergoing catheter ablation (n = 217) and (ii) a control group (n = 105). (i) Adipokines, oxidative stress, indirect autonomic markers, and leucocytes mRNA levels were analysed; (ii) correlation between biomarkers was explored with heatmaps and Kendall correlation coefficients; and (iii) logistic regression and random forest model were used to determine predictors of AF recurrence after ablation. Our results showed that: (i) fatty acid-binding protein 4 (FABP4) and leptin levels were higher in women than in men in both cohorts (P < 0.01). In women, FABP4 levels were higher on AF cohort (20 ± 14 control, 29 ± 18 paroxysmal AF and 31 ± 17 ng/mL persistent AF; P < 0.01). In men, leptin levels were lower on AF cohort (22 ± 15 control, 13 ± 16 paroxysmal AF and 13 ± 11 ng/mL persistent AF; P < 0.01). (ii) In female with paroxysmal AF, there was a lower acetylcholinesterase and higher carbonic anhydrase levels with respect to men (P < 0.05). (iii) Adipokines have an important role on discriminate AF recurrence after ablation. In persistent AF, FABP4 was the best predictor of recurrence after ablation (1.067, 95% confidence interval 1-1.14; P = 0.046). CONCLUSION: The major finding of the present study is the sex-based differences of FABP4 and leptin levels according to AF burden. These adipokines are associated with oxidative stress, inflammatory and autonomic indirect markers, indicating that they may play a role in AF perpetuation.

The use of dynamic prevalence maps to assess the prevalence of Salmonella in broiler flocks in the Valencian Community, Spain.

The aim of this study was to design and implement a dynamic prevalence map for managing the data collected within the framework of the Salmonella spp. National control plan in broiler flocks, using a data set from the Valencian Community, Spain. Such a map would be a useful tool for veterinarians seeking to define control options. The development of the mapping tool for this study involved three main phases: data collection, data analysis and data representation. The data used were obtained from the results of the National Control Programme (based on European Commission [EC] Regulation No. 2160/2003). The Salmonella prevalence data were represented on a map of the Valencian Community, which included geographical information on flock locations to facilitate the interpretation of the results from monitoring. The average prevalence of Salmonella spp. was 2.74% from 2009 to 2013. Moreover, the proportion of broiler flocks that remained positive for the target serovars according to EC Regulation No. 2160/2003 (S. Enteritidis and/orS. Typhimurium [including monophasic S. Typhimurium]) was less than 1%. Over the five-year period of the study (2009-2013), the area of Bajo Maestrazgo had the highest level of Salmonella spp. prevalence every year, thereby validating the usefulness of this mapping tool. The aim of the study is to detect geographical 'highpressure' Salmonella zones over time and make suggestions on where efforts should be focused in this region to reduce future Salmonella spp. prevalence. Local Veterinary Services could, using the map, identify the infection pressure of Salmonella in the geographical areas where farms are located. Proximity to higher prevalences may imply an increased risk of flock contamination. Although annual data are represented, data can be presented in real time with regular frequency (daily, weekly, monthly or quarterly) or even over periods of several years. Therefore, the dynamic prevalence map provides extremely useful epidemiological information to veterinarians on the prevention of Salmonella.

L'étude présentée par les auteurs visait à concevoir et mettre en application une carte dynamique de la prévalence afin de gérer les données collectées dans le cadre du plan national de contrôle des Salmonella spp. dans les cheptels de poulets de chair à partir des informations transmises par la Communauté valencienne (Espagne). L'objectif attendu était que cette carte puisse aider les vétérinaires à définir la meilleure stratégie de lutte. La conception du logiciel de cartographie s'est faite en trois étapes, portant respectivement sur la collecte, l'analyse et la représentation des données. Les données utilisées provenaient des résultats du Programme national de lutte (basé sur le Règlement [CE] n° 2160/2003 du Parlement européen et du Conseil). Les taux de prévalence de Salmonella ont été reportés sur une carte de la Communauté valencienne, qui contenait également des informations géographiques relatives à la localisation des élevages afin de faciliter l'interprétation des résultats du suivi. De 2009 à 2013, la prévalence moyenne de Salmonella spp. était de 2,74 %. En outre, le pourcentage de cheptels de poulets de chair restant positifs au regard des sérovars visés par le Règlement CE n° 2160/2003 (S. Enteritidis et/ou S. Typhimurium [y compris la souche monophasique de S. Typhimurium]) était inférieur à 1 %. Sur les cinq années de l'étude (2009­2013), la prévalence annuelle la plus élevée de Salmonella spp. restait concentrée dans la région de Bajo Maestrazgo, ce qui confirme l'utilité de cet outil cartographique. Le but de l'étude est de pouvoir détecter les zones géographiques à « forte pression ¼ de Salmonella dans le temps afin d'orienter en conséquence les efforts de lutte dans cette région et de réduire ainsi la prévalence des Salmonella spp. à l'avenir. Grâce à cette carte, les Services vétérinaires locaux pourront connaître la pression exercée par Salmonella dans les zones géographiques où se situent les élevages. La proximité des zones à forte prévalence peut représenter un risque accru de contamination des cheptels. L'outil propose par défaut une représentation annuelle des données mais celles-ci peuvent également être présentées en temps réel à intervalles réguliers (fréquence quotidienne, hebdomadaire, mensuelle ou trimestrielle), voire même sur des périodes couvrant plusieurs années. Ainsi, la carte dynamique des prévalences fournit aux vétérinaires des informations épidémiologiques extrêmement utiles pour la prévention de Salmonella.

Los autores describen un estudio encaminado a concebir y realizar un mapa dinámico de prevalencia para explotar los datos obtenidos como parte del plan nacional de lucha contra las salmonelas en bandadas de pollos asaderos, utilizando para ello un conjunto de datos correspondientes a la Comunidad Valenciana (España). Semejante mapa ofrecería a los veterinarios una herramienta útil para definir las fórmulas idóneas de lucha. La elaboración del dispositivo de cartografía para este estudio entrañaba tres grandes fases: la de obtención, la de análisis y la de representación de los datos. Los datos utilizados derivan de los resultados obtenidos con el Programa Nacional de Control (basado en el Reglamento de la Comisión Europea [CE] Nº 2160/2003). Para facilitar la interpretación de los resultados obtenidos con la labor de vigilancia, los datos de prevalencia de salmonelas fueron representados en un mapa de la Comunidad Valenciana que contenía información geográfica sobre la localización de las bandadas. De 2009 a 2013, la prevalencia media de Salmonella spp. fue del 2,74%. Por otra parte, la proporción de bandadas de pollos asaderos que seguían siendo positivas para las serovariantes estudiadas con arreglo al Reglamento (CE)Nº 2160/2003 (S. Enteritidis y/o S. Typhimurium [incluida S. Typhimurium monofásica]) era inferior al 1%. En el quinquenio cubierto por el estudio (2009­2013), la zona del Bajo Maestrazgo registró cada año la mayor prevalencia de salmonelas, hecho que confirmó la utilidad de esta herramienta cartográfica. El estudio tiene por objetivo determinar a lo largo del tiempo las zonas geográficas de 'alta presión' de salmonelas y hacer propuestas sobre las áreas de esa región en las que convendría centrar el trabajo para reducir en el futuro la prevalencia de salmonelas. Empleando el mapa, los Servicios Veterinarios podrían determinar la presión infecciosa de salmonelas en las zonas geográficas en que se encuentran las explotaciones. Cuanto más cerca esté una explotación de zonas con niveles altos de prevalencia, mayor puede ser el riesgo de contaminación de las bandadas. Aunque en el mapa se representan datos anuales, también es posible presentarlos en tiempo real con determinada periodicidad (diaria, semanal, mensual o trimestral) o incluso cubrir periodos de varios años. El mapa dinámico de prevalencia proporciona así a los veterinarios información epidemiológica de gran utilidad para la prevención de las salmonelosis.

Conformational and thermal characterization of left ventricle remodeling post-myocardial infarction.

Adverse cardiac remodeling after myocardial infarction (MI) causes impaired ventricular function and heart failure. Histopathological characterization is commonly used to detect the location, size and shape of MI sites. However, the information about chemical composition, physical structure and molecular mobility of peri- and infarct zones post-MI is rather limited. The main objective of this work was to explore the spatiotemporal biochemical and biophysical alterations of key cardiac components post-MI. The FTIR spectra of healthy and remote myocardial tissue shows amides A, I, II and III associated with proteins in freeze-died tissue as major absorptions bands. In infarcted myocardium, the spectrum of these main absorptions was deeply altered. FITR evidenced an increase of the amide A band and the distinct feature of the collagen specific absorption band at 1338cm-1 in the infarct area at 21days post-MI. At 21days post-MI, it also appears an important shift of amide I from 1646cm-1 to 1637cm-1 that suggests the predominance of the triple helical conformation in the proteins. The new spectra bands also indicate an increase in proteoglycans, residues of carbohydrates in proteins and polysaccharides in ischemic areas. Thermal analysis indicates a deep increase of unfreezable water/freezable water in peri- and infarcted tissues. In infarcted tissue is evidenced the impairment of myofibrillar proteins thermal profile and the emergence of a new structure. In conclusion, our results indicate a profound evolution of protein secondary structures in association with collagen deposition and reorganization of water involved in the scar maturation of peri- and infarct zones post-MI.

NAT2 gene diversity and its evolutionary trajectory in the Americas.

N-acetyltransferase 2 (NAT2) is responsible for metabolizing xenobiotics; NAT2 polymorphisms lead to three phenotypes: rapid, intermediate and slow acetylators. We aimed to investigate NAT2 diversity in Native Americans. NAT2 exon 2 was sequenced for 286 individuals from 21 populations (Native American and American Mestizos). Excluding the basal/rapid haplotype NAT2*4, the most frequent haplotypes are NAT2*5B (35.95%) in hunter-gatherers and NAT2*7B (20.61%) and NAT2*5B (19.08%) in agriculturalists that were related to the slow phenotype. A new haplotype was identified in two Amerindians. Data from the ~44 kb region surrounding NAT2 in 819 individuals from Africa, East-Asia, Europe and America were used in additional analyses. No significant differences in the acetylator NAT2 haplotype and phenotype distributions were found between Native American populations practicing farming and/or herding and those practicing hunting and gathering, probably because of the absence or weakness of selection pressures and presence of demographic and random processes preventing detection of any selection signal.

Ocular and craniofacial phenotypes in a large Brazilian family with congenital aniridia.

Congenital aniridia is a rare genetic disorder characterized by varying degrees of iris hypoplasia that are associated with additional ocular abnormalities. More than 90% of the causal mutations identified are found in the PAX6 gene, a transcription factor of critical importance in the process of neurogenesis and ocular development. Here, we investigate clinical, molecular, and craniofacial features of a large Brazilian family with congenital aniridia. Among the 56 eyes evaluated, phenotype variation encompassed bilateral total aniridia to mild iris defects with extensive variation between eyes of the same individual. PAX6 molecular screening indicated a heterozygous splice mutation (c.141 + 1G>A). Thus, we hypothesize that this splicing event may cause variation in the expression of the wild-type transcript, which may lead to the observed variation in phenotype. Affected individuals were more brachycephalic, even though their face height and cephalic circumference were not significantly different when compared to those of non-affected relatives. From this, we infer that the head shape of affected subjects may also be a result of the PAX6 splice-site mutation. Our data summarize the clinical variability associated with the ocular phenotype in a large family with aniridia, and help shed light on the role of PAX6 in neurocranial development.

Mechanisms modulating foam cell formation in the arterial intima: exploring new therapeutic opportunities in atherosclerosis.

In recent years, the role of macrophages as the primary cell type contributing to foam cell formation and atheroma plaque development has been widely acknowledged. However, it has been long recognized that diffuse intimal thickening (DIM), which precedes the formation of early fatty streaks in humans, primarily consists of lipid-loaded smooth muscle cells (SMCs) and their secreted proteoglycans. Recent studies have further supported the notion that SMCs constitute the majority of foam cells in advanced atherosclerotic plaques. Given that SMCs are a major component of the vascular wall, they serve as a significant source of microvesicles and exosomes, which have the potential to regulate the physiology of other vascular cells. Notably, more than half of the foam cells present in atherosclerotic lesions are of SMC origin. In this review, we describe several mechanisms underlying the formation of intimal foam-like cells in atherosclerotic plaques. Based on these mechanisms, we discuss novel therapeutic approaches that have been developed to regulate the generation of intimal foam-like cells. These innovative strategies hold promise for improving the management of atherosclerosis in the near future.

Cocoa consumption reduces NF-κB activation in peripheral blood mononuclear cells in humans.

BACKGROUND AND AIMS: Epidemiological studies have demonstrated an association between high-polyphenol intake and reduced incidence of atherosclerosis. The healthy effects of cocoa-polyphenols may be due to their antioxidant and anti-inflammatory actions, although the exact mechanisms are unknown and depend on the matrix in which cocoa-polyphenols are delivered. Nuclear factor κB (NF-κB) is a key molecule in the pathophysiology of atherosclerosis involved in the regulation of adhesion molecules(AM) and cytokine expression and its activation is the first step in triggering the inflammatory process. The aim of this study was to evaluate the effect of acute cocoa consumption in different matrices related to the bioavailability of cocoa-polyphenols in NF-κB activation and the expression of AM. METHODS AND RESULTS: Eighteen healthy volunteers randomly received 3 interventions: 40g of cocoa powder with milk (CM), with water (CW), and only milk (M). NF-κB activation in leukocytes and AM (sICAM, sVCAM, E-selectin) were measured before and 6h after each intervention. Consumption of CW significantly decreased NF-κB activation compared to baseline and to CM (P < 0.05, both), did not change after CM intervention, and significantly increased after M intervention (P = 0.014). sICAM-1 concentrations significantly decreased after 6h of CW and CM interventions (P ≤ 0.026; both) and E-selectin only decreased after CW intervention (P = 0.028). No significant changes were observed in sVCAM-1 concentrations. CONCLUSIONS: The anti-inflammatory effect of cocoa intake may depend on the bioavailability of bioactive compounds and may be mediated at least in part by the modulation of NF-κB activation and downstream molecules reinforcing the link between cocoa intake and health.

Molecular evolution of the melanocortin 1-receptor pigmentation gene in rodents.

Adaptive variation in the melanocortin 1-receptor gene (MC1R), a key locus in melanogenesis, has been identified in some species of rodents. However, in others, MC1R has no causative role in pigmentation phenotypes despite their coat color variation. In this study, we characterized the rates and patterns of MC1R nucleotide and amino acid sequence evolution and, particularly, selective pressures in the separated domains of the protein using a comparative analysis of 43 species representing three major lineages of rodents with variable coat colors. We found high amino acid variation (44% of sites) throughout the protein. Most substitutions were observed in extracellular and transmembrane domains; the intracellular segment was conserved across species. Pairwise non-synonymous substitutions did not vary significantly in different domains among the rodent lineages - i.e., variation was not associated with phylogenetic distance. Phylogeny-based likelihood analysis suggested that purifying selection has mostly shaped the evolutionary course of MC1R. However, a high proportion of sites (27%) were under relaxation of functional constraints (ω = 0.38), and four sites (3, 14, 26, and 251) clearly evolved under positive selection (ω ≅ 2.9). Thus, our data indicate a high proportion of sites evolving under relaxed evolutionary constraints, which might indicate the evolvability of the system in the generation of adaptive changes in specific taxa in rodent lineages.

Low density lipoprotein receptor-related protein 1 modulates the proliferation and migration of human hepatic stellate cells.

Human hepatic stellate cells (HHSCs) proliferation and migration play a key role in the pathogenesis of liver inflammation and fibrogenesis. Low density lipoprotein receptor-related protein (LRP1) is an endocytic receptor involved in intracellular signal transduction. The aim of this work was to analyse the role of low density lipoprotein receptor-related protein (LRP1) in HHSCs proliferation and migration and the mechanisms involved. Human LRP1 deficient-HHSCs were generated by nucleofecting the line HHSCs with siRNA anti-LRP1. HHSCs DNA synthesis was measured by [(3) H]-thymidine incorporation and cell cycle progression by flow cytometry after annexin V and iodure propidium staining. Cell migration was assessed using a wound repair model system. LRP1 expression and extracellular matrix-regulated kinase (ERK1,2) phosphorylation were analysed by Western blot analysis. Transforming growth factor-ß (TGF-ß) extracellular levels were analysed by ELISA. siRNA-antiLRP1 treatment almost completely inhibited LRP1 mRNA and protein expression. LRP1 deficient HHSCs showed higher proliferative response (172 ± 19 vs. 93 ± 8 [(3) H]-thymidine incorporation; 78.68% vs. 82.69% in G0/G1, 21.32% vs. 17.30% in G2/S) and higher migration rates than control HHSCs. LRP1 deficient cells showed higher levels of phosphorylated ERK1,2. TGF-ß extracellular levels were threefold higher in LRP1-deficient than in control HHSCs cells. These results demonstrate that LRP1 regulates HHSCs proliferation and migration through modulation of ERK1,2 phosphorylation and TGF-ß extracellular levels. These results suggest that HHSCs-LRP1 may play a key role in the modulation of factors determining hepatic fibrosis.

Heuristic optimization for the design of plasmonic nanowires with specific resonant and scattering properties.

In this contribution, we propose a computational tool for the synthesis of metallic nanowires with optimized optical properties, e.g. maximal scattering cross-section at a given wavelength. For this, we employ a rigorous numerical method, based on the solution of surface integral equations, along with a heuristic optimization technique that belongs to the population-based set known as Evolutionary Algorithms. Also, we make use of a general representation scheme to model, in a more realistic manner, the arbitrary geometry of the nanowires. The performance of this approach is evaluated through some examples involving various wavelengths, materials, and optimization strategies. The results of our numerical experiments show that this hybrid technique is a suitable and versatile tool straightforwardly extensible for the design of different configurations of interest in Plasmonics.

New vascular structure in broiler breast: breast-vessel.

Chickens intensively selected for accelerating growth rate and enhanced muscle mass has resulted in the occurrence of breast alterations with a negative consumers perception. Together with the already known breast abnormalities (White Striping, Wooden Breast and Spaghetti Meat) a new one has been described by the consumers as a "long and thin worm". For this reason, the aim of this work was to macroscopically and microscopically describe this structure in broiler breast. To this end, 2 different experiments were performed. In the first one, 4 broiler breasts from a supermarket were analysed. Thereafter, a second experiment was carried out on an experimental farm simulating field management conditions in order to confirm the results of the first experiment. To this end, 120 chickens were reared in an experimental farm up to 42 d. Breast samples were collected and analysed macroscopically and microscopically to assess the abnormality on the broiler breast. In both experiments, the results of the analysis showed the finding of tubular structures located on the surface of pectoralis major muscle compatible with arteries. So far, currently, this occurrence has not been described in the literature and we propose calling this new broiler breast feature as breast-vessel.

Monitoring serologic response to single in ovo vaccination with an immune complex vaccine against infectious bursal disease in broilers.

The infectious bursal disease (IBD) virus is one of the most resistant and prevalent virus worldwide in the poultry industry, being vaccination the main tool to control the disease. For this reason, consistent and uniform immunization of broiler flocks against IBD is necessary to avoid the disease spreading. The aim of this study was to apply and assess an epidemiologic mapping tool focused on the immunization by in ovo single broiler vaccination using an immune complex IBD vaccine. With this regard, 7,576 serum samples were collected from 603 broiler flocks raised in 354 Spanish farms. To do so, blood samples were randomly collected from birds with ages between 35 to 51 d, and the serum was analyzed by ELISA. The results obtained from this study suggested a high uniform immunization against IBDV and a protective immunization between 35 and 51 d of age, with mean titer values ranging between 6,331 and 7,426. In addition, seroprevalence titer data of this large-scale monitoring study fitted a polynomial equation with a R2 value of 0.77, helping to explain and predict the humoral response to IBD vaccination. This seroprevalence map was applied to broiler production and was based on business intelligence tool that incorporates newly developed mapping tool to cover the need of having real-time information of humoral response to IBD vaccination and could be an effective tool for veterinary services to control and prevent IBD.

Salmonella Infantis and Salmonella Enteritidis specific bacteriophages isolated form poultry faeces as a complementary tool for cleaning and disinfection against Salmonella.

Salmonellosis represents an important public health concern. Several authors point out the inefficiency of the cleaning and disinfection protocols to remove the bacteria from the field. For this reason, innovative techniques, as bacteriophages, could be implemented to control the bacteria. The main objectives of this study were to assess the effect of bacteriophages against Salmonella Infantis and Salmonella Enteritidis on farm surfaces, and to evaluate bacteriophage procedure application as sanitiser against Salmonella in field conditions. Thus, most prevalent serovars in poultry production were selected (Salmonella Infantis and Salmonella Enteritidis) to contaminate farm facilities. Then, two specific bacteriophages isolated from poultry faeces were applied against them. Results showed Salmonella Infantis and Salmonella Enteritidis decreased of 4.55 log10CFU/mL and 3.85 log10CFU/mL, respectively; the maximum reduction in Salmonella was the 5th day, after 108 PFU/mL and 103 PFU/mL bacteriophage application. These results highlight bacteriophages as a promising tool together with cleaning and disinfection.

Feather mites and birds: an interaction mediated by uropygial gland size?

Feather mites (Arachnida: Acari: Astigmata) feed mainly on secretions of the uropygial gland of birds. Here, we use analyses corrected for phylogeny and body size to show that there is a positive correlation between the size of this gland and mite abundance in passerine birds at an interspecific level during the breeding season, suggesting that the gland mediates interactions between mites and birds. As predicted on the basis of hypothesized waterproofing and antibiotic functions of uropygial gland secretions, riparian/marsh bird species had larger glands and higher mite loads than birds living in less mesic terrestrial environments. An unexpected pattern was a steeper relationship between mite load and gland size in migratory birds than in residents. If moderate mite loads are beneficial to a host but high loads detrimental, this could create complex selection regimes in which gland size influences mite load and vice versa. Mites may exert selective pressures on gland size of their hosts that has resulted in smaller glands among migratory bird species, suggesting that smaller glands may have evolved in these birds to attenuate a possible detrimental effect of feather mites when present in large numbers.

Tissue factor induction by aggregated LDL depends on LDL receptor-related protein expression (LRP1) and Rho A translocation in human vascular smooth muscle cells.

OBJECTIVE: Low density lipoprotein (LDL) internalized in the vascular wall and modified by binding to extracellular matrix-proteoglycans (ECM) becomes aggregated (agLDL). AgLDL induces tissue factor (TF) expression and activity in human vascular smooth muscle cells (VSMC). TF expression in vascular cells promotes the prothrombotic transformation of the vascular wall. However, the mechanisms by which agLDL induces TF are not known. The aim of this study was to investigate the mechanisms involved in TF activation by extracellular matrix-modified LDL in human VSMC. METHODS AND RESULTS: AgLDL significantly induces TF expression (real time PCR and Western blot analysis) and procoagulant activity (factor Xa generation test) in human VSMC. HMG-CoA reductase inhibition completely prevents agLDL-induced TF expression and partially inhibits agLDL-TF activation. These effects are reverted by geranylgeranyl pyrophosphate (GGPP) but not by farnesyl pyrophosphate (FPP), suggesting the involvement of a geranylated protein in agLDL-TF induction. AgLDL increases Rho A translocation (2-fold) from the cytoplasm to the cell membrane in control but not in simvastatin-treated VSMC. Exoenzyme C3, a specific Rho A inhibitor, completely prevents agLDL-induced TF overexpression and partially agLDL-TF activation. Blocking LRP1, the receptor of agLDL, with anti-LRP1 antibodies or inhibiting LRP1 expression by small interference RNA treatment (siRNA-LRP1) impairs agLDL-induced TF overexpression and activation. CONCLUSIONS: These results demonstrate that TF induction by agLDL depends on LRP1 expression and requires Rho A translocation to the cellular membrane.

Autophage as a control measure for Salmonella in laying hens.

Notwithstanding the National Salmonella Control Programs, the latest data published by the European Food Safety Authority show an increase in Salmonella Enteritidis prevalence in laying hen flocks. For this reason, the implementation of innovative techniques such as phage therapy is needed to control Salmonella at farm level. Most common bacteriophage applications are a cocktail of 2 or more bacteriophages, as it has been described that cocktails could remove different Salmonella serotypes, thus providing cross efficacy. Nevertheless, resistance to the bacteriophage cocktail has been reported, resulting in a decrease in their effectiveness. Along these lines, some authors have reported the possibility of using autophage when commercial bacteriophage cocktails are not active against field strains. To our best knowledge, no autophage (bacteriophage isolated from the same environment where the pathogen is isolated) has been found to control Salmonella in laying hens. In this context, the aim of this study was to assess the application of autophage in reducing Salmonella Enteritidis in environmental and fecal samples in a layer farm. To this end, the bacteriophage was isolated from the same farm where the bacteria was present and was applied onto the facility installations and the animals, at 2 different times. After bacteriophage challenges, swab cloths from facility surfaces and feces samples were collected at 3 times according to the time spent after the bacteriophage challenge. The results obtained in our study showed that all the surface samples collected from the farm facilities after phage therapy were negative for Salmonella. Concerning faces samples, statistical differences were found in Salmonella counts, with the strongest decrease (1.78log10) occurring after the second challenge. Otherwise, depending of the moment of sampling, the results obtained were 2.34log10, 1.39log10, 0.56log10, and 0.97log10 cfu/g for T0, T1, T2, and T3 respectively. The study highlights the use of autophage therapy not only for Salmonella Enteritidis control in animals, but as a sanitizer in cleaning and disinfection.

Sterol regulatory element-binding protein-2 negatively regulates low density lipoprotein receptor-related protein transcription.

Low density lipoprotein receptor-related protein (LRP1) binds aggregated LDL (agLDL) leading to a high intracellular cholesteryl ester (CE) accumulation. AgLDL up-regulates LRP1 expression concomitantly with an LDL receptor (LDLR) and sterol regulatory element binding protein (SREBP-2) down-regulation. The objectives were to investigate whether SREBP-2 regulates LRP1 transcription and determine the molecular mechanisms involved in the process. Down-regulation of active SREBP-2 by nLDL and agLDL led to LDLR down-regulation and LRP1 up-regulation. Enforced expression of an active form of SREBP-2 (SREBP-2-NT, amino acid residues 1-468) decreased LRP1 expression and LRP1 promoter (WT-LRP1) luciferase activity in a dose-dependent manner. LDL did not exert any significant effect on LRP1 promoter activity when a putative sterol regulatory element (SRE) (5-GTGGGGTGA-3'; +225 to +233) was mutated (SRE-MT-LRP1). SREBP-2 overexpression exerted stronger down-regulatory effects on WT-LRP1 than on SRE-MT-LRP1 promoter activity both in control, nLDL- and agLDL-exposed HeLa cells. Gel mobility shift assays showed that recombinant SREBP-2-NT protein (1-468) binds to a double-stranded LRP1 DNA fragment (215 to 245) containing a wild-type (wt) SRE sequence but not to a mutated SRE (mt) sequence (5-GAATTCGA-3'). Our results demonstrate that LDL stimulates LRP1 transcription and decreases SREBP-2 active form which negatively regulates LRP1 transcription. SRE sequence (+225 to +233) plays a pivotal role for the down-regulatory effect of SREBP-2 on LRP1 promoter activity.

Cell biology and lipoproteins in atherosclerosis.

Atherosclerosis is an inflammatory process, triggered by the presence of lipids in the vascular wall, and encompasses a complex interaction among inflammatory cells, vascular elements, and lipoproteins through expression of several adhesion molecules and cytokines. Subendothelial retention of lipoproteins is the key initiating event in atherosclerosis, provoking a cascade of events to pathogenic response. High levels of plasma lipids, particularly low-density (LDL) and very-low-density lipoproteins (VLDL) are among the pathophysiologic stimuli that induce endothelial dysfunction. Endothelial cells regulate coagulation, thrombosis and the fibrinolytic system; the endothelium modulates the activity of smooth muscle cells (vascular tone/proliferation) and controls the traffic of macromolecules and inflammatory cells to the vessel wall. Furthermore, LDLs have been implicated in the induction of changes in permeability, cell adhesion and secretion of vasoactive molecules (nitric oxide [NO]), while VLDLs seem to modulate the fibrinolytic system [tissue plasminogen activator (TPA) and plasminogen activator inhibitor-1 (PAI-1)]. In this review, we will focus on the pathophysiologic functions of lipoproteins in the vascular wall.

Preoperative progressive pneumoperitoneum and botulinum toxin type A in patients with large incisional hernia.

PURPOSE: Combination of preoperative progressive pneumoperitoneum (PPP) and botulinum toxin type A (BT) has not been previously reported in the management of large incisional hernia (LIH). METHODS: Observational study of 45 consecutive patients with LIH between June 2010 and July 2014. The diameters of the hernia sac, the volumes of the incisional hernia (VIH) and the abdominal cavity (VAC), and the VIH/VAC ratio were measured before and after PPP and BT using abdominal CT scan data. We indicated the combination of both techniques when the volume of the incisional hernia (VIH)/volume of the abdominal cavity (VAC) ratio was >20%. RESULTS: The median insufflated volume of air for PPP was 8.600 ± 3.200 cc (4.500-13.250), over a period of 14.3 ± 1.3 days (13-16). BT administration time was 40.2 ± 3.3 days (37-44). We obtained an average value of reduction of 14% of the VIH/VAC ratio after PPP and BT (p < 0.05). Complications associated with PPP were 15.5%, and with surgical technique, 26.6%. No complications occurred during the BT administration. Reconstructive technique was anterior CST and primary fascial closure was achieved in all patients. Median follow-up was 40.5 ± 19 months (12-60) and we reported 2 cases of hernia recurrence (4.4%). CONCLUSIONS: Preoperative combination of PPP and BT is feasible and a useful tool in the surgical management of LIH, although at the cost of some specific complications.

Familial dilated cardiomyopathy: A multidisciplinary entity, from basic screening to novel circulating biomarkers.

Idiopathic dilated cardiomyopathy has become one of the most prevalent inherited cardiomyopathies over the past decades. Genetic screening of first-degree relatives has revealed that 30-50% of the cases have a familial origin. Similar to other heart diseases, familial dilated cardiomyopathy is characterized by a high genetic heterogeneity that complicates family studies. Cli'nical screening, 12-lead electrocardiogram and transthoracic echocardiogram are recommended for patients and first-degree family members. Magnetic resonance also needs to be considered. Genetic technologies have become fundamental for the clinical management of this disease. New generation sequencing methods have made genetic testing feasible for extensive panels of genes related to the disease. Recently, new imaging modalities such as speckle-tracking, strain and strain rate or magnetic resonance, and circulating biomarkers such as non-coding RNAs, have emerged as potential strategies to help cardiologists in their clinical practice. Imaging, genetic and blood-based techniques should be considered together in the evaluation and testing of familial dilated cardiomyopathy. Here, we discuss the current procedures and novel approaches for the clinical management of familial dilated cardiomyopathy.