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Legal initiatives recognizing the rights of nature have transformed from a trickle to a cascade. But are they really effective? A systematic study on the implementation of the Ecuadorian Los Cedros ruling, one of the most prominent decisions of this sort, shows that it has been notably effective in protecting the forest from mining threats. However, its impact on local community involvement has been limited, and the fate of Los Cedros and the case remain vulnerable.

Clostridioides difficile is an emerging pathogen of One Health significance. Its highly variable genome contains mobile genetic elements (MGEs) such as transposons and prophages that influence its biology. Systematic deletion of each genetic element is required to determine their precise role in C. difficile biology and contribution to the wider mobilome. Here, Tn5397 (21 kb) and ϕ027 (56 kb) were deleted from C. difficile 630 and R20291, respectively, using allele replacement facilitated by CRISPR-Cas9. The 630 Tn5397 deletant transferred PaLoc at the same frequency (1 × 10−7) as 630 harboring Tn5397, indicating that Tn5397 alone did not mediate conjugative transfer of PaLoc. The R20291 ϕ027 deletant was sensitive to ϕ027 infection, and contained two unexpected features, a 2.7 kb remnant of the mutagenesis plasmid, and a putative catalase gene adjacent to the deleted prophage was also deleted. Growth kinetics of R20291 ϕ027 deletant was similar to wild type (WT) in rich medium but marginally reduced compared with WT in minimal medium. This work indicates the commonly used pMTL8000 plasmid series works well for CRISPR-Cas9-mediated gene deletion, resulting in the largest deleted locus (56.8 kb) described in C. difficile. Removal of MGEs was achieved by targeting conjugative/integrative regions to promote excision and permanent loss. The deletants created will be useful strains for investigating Tn5397 or ϕ027 prophage contribution to host virulence, fitness, and physiology, and a platform for other mutagenesis studies aimed at functional gene analysis without native transposon or phage interference in C. difficile 630 and R20291.

(1) Background: Given the widespread use of Alkalihalobacillus clausii (A. clausii) as a probiotic in recent decades and the detection of bacteremia cases in a group of patients, we sought to analyze cases of A. clausii bacteremia following oral probiotic use (2) Methods: A retrospective observational study was conducted at a private hospital in San Jose, Costa Rica. Cases of bacteremia caused by A. clausii confirmed by the microbiology laboratory were analyzed in patients who received oral treatment with this probiotic between January 2020 and January 2022. In addition, an isolate (HCB-AC2) was compared through whole genome sequencing to demonstrate the correlation of bacteremia and A. clausii. Possible vulnerability factors related to the development of this condition were determined. (3) Results: Four cases were identified in this hospital over 2 years. Genomic analysis of isolate HCB-AC2, using two different methods, showed identical results. This indicates that HCB-AC2 is genomically identical to ENTpro and the Enterogermina® reference genome. The median age was 71 years, and all patients had some degree of immunosuppression. All patients met at least three sepsis criteria at the time of bacterial identification. Most patients were treated with vancomycin and levofloxacin. Three of the identified patients died. (4) Conclusion: A. clausii can be used as a probiotic, but caution is advised when used in immunosuppressed and elderly patients. These findings align with those reported in similar case studies.

This study addresses the importance of conducting mass movement susceptibility mapping and hazard assessment using quantitative techniques, including machine learning, in the Northern Lima Commonwealth (NLC). A previous exploration of the topographic variables revealed a high correlation and multicollinearity among some of them, which led to dimensionality reduction through a principal component analysis (PCA). Six susceptibility models were generated using weights of evidence, logistic regression, multilayer perceptron, support vector machine, random forest, and naive Bayes methods to produce quantitative susceptibility maps and assess the hazard associated with two scenarios: the first being El Niño phenomenon and the second being an earthquake exceeding 8.8 Mw. The main findings indicate that machine learning models exhibit excellent predictive performance for the presence and absence of mass movement events, as all models surpassed an AUC value of >0.9, with the random forest model standing out. In terms of hazard levels, in the event of an El Niño phenomenon or an earthquake exceeding 8.8 Mw, approximately 40% and 35% respectively, of the NLC area would be exposed to the highest hazard levels. The importance of integrating methodologies in mass movement susceptibility models is also emphasized; these methodologies include the correlation analysis, multicollinearity assessment, dimensionality reduction of variables, and coupling statistical models with machine learning models to improve the predictive accuracy of machine learning models. The findings of this research are expected to serve as a supportive tool for land managers in formulating effective disaster prevention and risk reduction strategies.

The inhibition exerted by ethanolic extracts of coffee mucilage on the growth of bacteria was studied by microdilution in agar. The growth inhibition effect was evaluated for pathogenic or food spoilage related bacteria (Pseudomonas aeruginosa (ATCC 27853), Alcaligenes sp. (UCR277), Serratia sp. (UCR299), Micrococcus luteus (ATCC4698), Escherichia coli (ATCC35150), Staphylococcus aureus (ATCC25923), Bacillus cereus (ATCC14579), Salmonella enterica subsp. enterica (ATCC 13311) and Listeria monocytogenes (SLCC4013)) and for bacteria associated with human intestinal biota (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactiplantibacillus plantarum subsp. plantarum). The most effective growth inhibition was observed for B. cereus (ATCC14579). The content of chlorogenic acid and caffeine in the ethanolic extracts was quantified by HPLC/DAD. The chlorogenic acid content in the extracts ranged from 2.67 to 4.76 mg/ml, while the caffeine content ranged from 1.24 to 6.48 mg/ml. Although ethanolic extracts of coffee berry mucilage inhibited the growth of B. cereus, this inhibition does not seem to be related to the caffeine or chlorogenic acid contents.

ABSTRACTThe intestinal pathogen Clostridioides difficile is the leading cause of antibiotic-associated diarrhea and pseudomembranous colitis in humans. The symptoms of C. difficile-associated diseases (CDADs) are directly associated with the pathogen’s toxins TcdA and TcdB, which enter host cells and inactivate Rho and/or Ras GTPases by glucosylation. Membrane cholesterol is crucial during the intoxication process of TcdA and TcdB, and likely involved during pore formation of both toxins in endosomal membranes, a key step after cellular uptake for the translocation of the glucosyltransferase domain of both toxins from endosomes into the host cell cytosol. The licensed drug amiodarone, a multichannel blocker commonly used in the treatment of cardiac dysrhythmias, is also capable of inhibiting endosomal acidification and, as shown recently, cholesterol biosynthesis. Thus, we were keen to investigate in vitro with cultured cells and human intestinal organoids, whether amiodarone preincubation protects from TcdA and/or TcdB intoxication. Amiodarone conferred protection against both toxins independently and in combination as well as against toxin variants from the clinically relevant, epidemic C. difficile strain NAP1/027. Further mechanistic studies suggested that amiodarone’s mode-of-inhibition involves also interference with the translocation pore of both toxins. Our study opens the possibility of repurposing the licensed drug amiodarone as a novel pan-variant antitoxin therapeutic in the context of CDADs.

Abstract Toxin B (TcdB) is a major exotoxin responsible for diseases associated with Clostridioides difficile infection. Its sequence variations among clinical isolates may contribute to the difficulty in developing effective therapeutics. Here, we investigate receptor-binding specificity of major TcdB subtypes (TcdB1 to TcdB12). We find that representative members of subtypes 2, 4, 7, 10, 11, and 12 do not recognize the established host receptor, frizzled proteins (FZDs). Using a genome-wide CRISPR-Cas9-mediated screen, we identify tissue factor pathway inhibitor (TFPI) as a host receptor for TcdB4. TFPI is recognized by a region in TcdB4 that is homologous to the FZD-binding site in TcdB1. Analysis of 206 TcdB variant sequences reveals a set of six residues within this receptor-binding site that defines a TFPI binding-associated haplotype (designated B4/B7) that is present in all TcdB4 members, a subset of TcdB7, and one member of TcdB2. Intragenic micro-recombination (IR) events have occurred around this receptor-binding region in TcdB7 and TcdB2 members, resulting in either TFPI- or FZD-binding capabilities. Introduction of B4/B7-haplotype residues into TcdB1 enables dual recognition of TFPI and FZDs. Finally, TcdB10 also recognizes TFPI, although it does not belong to the B4/B7 haplotype, and shows species selectivity: it recognizes TFPI of chicken and to a lesser degree mouse, but not human, dog, or cattle versions. These findings identify TFPI as a TcdB receptor and reveal IR-driven changes on receptor-specificity among TcdB variants.

The study, synthesis, and application of nanomaterials in medicine have grown exponentially in recent years. An example of this is the understanding of how nanomaterials activate or regulate the immune system, particularly macrophages. In this work, nanoparticles were synthesized using Rumex hymenosepalus as a reducing agent (AgRhNPs). According to thermogravimetric analysis, the metal content of nanoparticles is 55.5% by weight. The size of the particles ranges from 5–26 nm, with an average of 11 nm, and they possess an fcc crystalline structure. The presence of extract molecules on the nanomaterial was confirmed by UV-Vis and FTIR. It was found by UPLC-qTOF that the most abundant compounds in Rh extract are flavonols, flavones, isoflavones, chalcones, and anthocyanidins. The viability and apoptosis of the THP-1 cell line were evaluated for AgRhNPs, commercial nanoparticles (AgCNPs), and Rh extract. The results indicate a minimal cytotoxic and apoptotic effect at a concentration of 12.5 μg/mL for both nanoparticles and 25 μg/mL for Rh extract. The interaction of the THP-1 cell line and treatments was used to evaluate the polarization of monocyte subsets in conjunction with an evaluation of CCR2, Tie-2, and Arg-1 expression. The AgRhNPs nanoparticles and Rh extract neither exhibited cytotoxicity in the THP-1 monocyte cell line. Additionally, the treatments mentioned above exhibited anti-inflammatory effects by maintaining the classical monocyte phenotype CD14++CD16, reducing pro-inflammatory interleukin IL-6 production, and increasing IL-4 production.

Antioxidant defenses in biological systems ensure redox homeostasis, regulating baseline levels of reactive oxygen and nitrogen species (ROS and RNS). Oxidative stress (OS), characterized by a lack of antioxidant defenses or an elevation in ROS and RNS, may cause a modification of biomolecules, ROS being primarily absorbed by proteins. As a result of both genome and environment interactions, proteomics provides complete information about a cell’s proteome, which changes continuously. Besides measuring protein expression levels, proteomics can also be used to identify protein modifications, localizations, the effects of added agents, and the interactions between proteins. Several oxidative processes are frequently used to modify proteins post-translationally, including carbonylation, oxidation of amino acid side chains, glycation, or lipid peroxidation, which produces highly reactive alkenals. Reactive alkenals, such as 4-hydroxy-2-nonenal, are added to cysteine (Cys), lysine (Lys), or histidine (His) residues by a Michael addition, and tyrosine (Tyr) residues are nitrated and Cys residues are nitrosylated by a Michael addition. Oxidative and nitrosative stress have been implicated in many neurodegenerative diseases as a result of oxidative damage to the brain, which may be especially vulnerable due to the large consumption of dioxygen. Therefore, the current methods applied for the detection, identification, and quantification in redox proteomics are of great interest. This review describes the main protein modifications classified as chemical reactions. Finally, we discuss the importance of redox proteomics to health and describe the analytical methods used in redox proteomics.

El siguiente estudio de investigación analizará de manera pormenorizada las aplicaciones de un bordado de alta costura presentes en el vestido de color rosa que Audrey Hepburn utilizó para la película Breakfast at Tiffany’s (1961). Basándose tanto en el cambio cromático del guipur al igual que la fragilidad sufrida sobre el bordado aplicado, su estudio científico intentará esclarecer la fuente de degradación activa, así como, la composición química de los bordados aplicados. Además, su caracterización aportará material bibliográfico nuevo sobre la producción de los bordados en alta costura de la década de los 60. Por último, a través del estudio de los archivos y la consulta de documentación se conseguirá ofrecer por primera vez, el lugar de producción del bordado para la colección de otoño-invierno de 1960, de la cual Hubert de Givenchy se inspiró para la realización del vestido rosa para Breakfast at Tiffany´s.

Abstract River barriers affect river dynamics and aquatic biota, altering the entire ecosystem. Nevertheless, dams and reservoirs provide goods like water supply and low-carbon energy that are becoming increasingly critical under current climate change. To know to what extent dams and reservoirs are important to the population, we explored social attitudes towards dams and reservoirs using a face-to-face questionnaire in two regions of contrasting climate and water security in Spain, a country with one of highest densities of dams in Europe. Results (N = 613) revealed a higher support for dams, mediated by the recognition of the services they provide, in the drier Mediterranean Malaga province (Andalusia), than in the wetter Atlantic Asturias province (Bay of Biscay), where water shortages are rare. Awareness of the impacts of the dams was more pronounced in Malaga, coupled with a higher willingness to pay for reconnecting rivers. Social awareness of both impacts and services provided by dams and reservoirs may depend on local climate and water security; different dam acceptance emphasizes the need to involve local citizens in the decision-making processes about water management.

The circadian clock is a regulatory system, with a periodicity of approximately 24 h, which generates rhythmic changes in many physiological processes, including mitochondrial activity. Increasing evidence links chronodisruption with aberrant functionality in clock gene expression, resulting in multiple diseases such as cancer. Melatonin, whose production and secretion oscillates according to the light–dark cycle, is the principal regulator of clock gene expression. In addition, the oncostatic effects of melatonin correlate with an increase in mitochondrial activity. However, the direct links between circadian clock gene expression, mitochondrial activity, and the antiproliferative effects of melatonin in cancers, including head and neck squamous cell carcinoma (HNSCC), remain largely unknown. In this study, we analyzed the effects of melatonin on HNSCC cell lines (Cal-27 and SCC9), which were treated with 500 and 1000 µM melatonin. We found that the antiproliferative effect of melatonin is not mediated by the Bmal1 clock gene. Additionally, high doses of melatonin were observed to result in resynchronization of oscillatory circadian rhythm genes (Per2 and Sirt1). Surprisingly, the resynchronizing effect of melatonin on Per2 and Sirt1 did not produce alterations in the oscillation of mitochondrial respiratory activity. These results increase our understanding of the possible antiproliferative mechanisms in melatonin in the treatment of head and neck squamous cell carcinoma and suggest that its antiproliferative effects are independent of clock genes but are directly related to mitochondrial activity.

An operando DRIFT-MS system (Diffuse Reflectance Infrared Fourier Transform Spectroscopy coupled with Mass Spectrometry) was designed and set up to study the oxidative steam reforming of ethanol reaction (OSRE). This reaction involves the mixture of water, ethanol and oxygen to produce mainly hydrogen, which is a rather attractive energy carrier. Spectroscopic monitoring of the process is a key tool to contribute to the understanding of: i) the dynamics on the catalyst surface, ii) the reaction mechanism and iii) the effect of the solid's properties on the catalytic process. In this sense, this document sets forth the experimental design that allows to carry out the study under operando DRIFT-MS conditions through time for the OSRE reaction. Selection criteria for parameters, materials, configuration, and experimental conditions are included, particularly optimizing the parameters of particle size and the dilution factor with KBr as well as the temperature and flow conditions for carrying out the reaction. • Clear signals of the adsorbed species in IR that do not present interference by water in the reaction atmosphere. • Simple assembly and online product detection by MS that allow to follow the change in the products of the OSRE reaction according to the temperature. • Controlled entry of gases and quantification by loop injection.

Abstract In this work, we used a sequential method of synthesis for gold–silver bimetallic nanoparticles with core@shell structure (Au@AgNPs). Rumex hymenosepalus root extract (Rh), which presents high content in catechins and stilbenes, was used as reductor agent in nanoparticles synthesis. Size distribution obtained by Transmission Electron Microscopy (TEM) gives a mean diameter of 36 ± 11 nm for Au@AgNPs, 24 ± 4 nm for gold nanoparticles (AuNPs), and 13 ± 3 nm for silver nanoparticles (AgNPs). The geometrical shapes of NPs were principally quasi-spherical. The thickness of the silver shell over AuNPs is around 6 nm and covered by active biomolecules onto the surface. Nanoparticles characterization included high angle annular dark field images (HAADF) recorded with a scanning transmission electron microscope (STEM), Energy-Dispersive X-ray Spectroscopy (EDS), X-Ray Diffraction (XRD), UV–Vis Spectroscopy, Zeta Potential, and Dynamic Light Scattering (DLS). Fourier Transform Infrared Spectrometer (FTIR), and X-ray Photoelectron Spectroscopy (XPS) show that nanoparticles are stabilized by extract molecules. A growth kinetics study was performed using the Gompertz model for microorganisms exposed to nanomaterials. The results indicate that AgNPs and Au@AgNPs affect the lag phase and growth rate of Escherichia coli and Candida albicans in a dose-dependent manner, with a better response for Au@AgNPs

Este artículo aborda la cuestión del estudio metafísico de la vida y lo vivo, partiendo a modo de guía de las nociones que Zubiri expone en su libro Estructura Dinámica de la Realidad, en el que el viviente es descubierto como «realidad en mismidad». Se acompaña el análisis de algunos ejemplos tomados de literatura científica y de comentarios y actualizaciones filosóficas de las tesis zubirianas. Además, se dejan meridianamente perfiladas algunas cuestiones para su estudio en la incipiente disciplina filosófica de la reología.

Clostridium perfringens is the causative agent of many enterotoxic diseases in humans and animals, and it is present in diverse environments (soil, food, sewage, and water). Multilocus Sequence Typing (MLST) and Whole Genome Sequencing (WGS) have provided a general approach about genetic diversity of C. perfringens; however, those studies are limited to specific locations and often include a reduced number of genomes. In this study, 372 C. perfringens genomes from multiple locations and sources were used to assess the genetic diversity and phylogenetic relatedness of this pathogen. In silico MLST was used for typing the isolates, and the resulting sequence types (ST) were assigned to clonal complexes (CC) based on allelic profiles that differ from its founder by up to double-locus variants. A pangenome analysis was conducted, and a core genome-based phylogenetic tree was created to define phylogenetic groups. Additionally, key virulence factors, toxinotypes, and antibiotic resistance genes were identified using ABRicate against Virulence Factor Database (VFDB), TOXiper, and Resfinder, respectively. The majority of the C. perfringens genomes found in publicly available databases were derived from food (n = 85) and bird (n = 85) isolates. A total of 195 STs, some of them shared between sources such as food and human, horses and dogs, and environment and birds, were grouped in 25 CC and distributed along five phylogenetic groups. Fifty-three percent of the genomes were allocated to toxinotype A, followed by F (32%) and G (7%). The most frequently found virulence factors based on > 70% coverage and 99.95% identity were plc (100%), nanH (99%), ccp (99%), and colA (98%), which encode an alpha-toxin, a sialidase, an alpha-clostripain, and a collagenase, respectively, while tetA (39.5%) and tetB (36.2%), which mediate tetracycline resistance determinants, were the most common antibiotic resistance genes detected. The analyses conducted here showed a better view of the presence of this pathogen across several host species. They also confirm that the genetic diversity of C. perfringens is based on a large number of virulence factors that vary among phylogroups, and antibiotic resistance markers, especially to tetracyclines, aminoglycosides, and macrolides. Those characteristics highlight the importance of C. perfringens as a one of the most common causes of foodborne illness.

BackgroundMigrants in Mexico are entitled to care at all levels, independently of their migration status. However, previous studies show that access to care is difficult for this population. As the movement of in-transit migrants and asylum seekers has been interrupted at the Mexico-United States border by migration policies such as the “Remain in Mexico” program, and by border closures due to the COVID-19 pandemic, the Mexican health system has the challenge of providing them with health care. Levesque et al.'s framework, according to which access occurs at the interface of health system characteristics and potential users' abilities to interact with it, is a useful theoretical tool to analyze the barriers faced by migrants.ObjectiveThe objective of this article is to analyze the barriers to access the public Mexican health system, encountered by migrants in cities in Mexican states at the Mexico-United States border during the COVID-19 pandemic.MethodsData came from a multiple case study of the response of migrant shelters to health care needs during the COVID-19 pandemic. The study consisted of a non-probability survey of migrants with a recent health need, and interviews with persons working in civil society organizations providing services to migrants, governmental actors involved in the response to migration, and academics with expertise in the subject. We analyzed the quantitative and qualitative results according to Levesque et al.'s framework.Results36/189 migrants surveyed had sought health care in a public service. The main limitations to access were in the availability and accommodation dimension (administrative barriers decreasing migrants' ability to reach the system), and the affordability dimension (out-of-pocket costs limiting migrants' ability to pay). Civil society organizations were a major source of social support, helping migrants overcome some of the barriers identified.ConclusionsWhile Mexico's health regulations are inclusive of migrants, in practice there are major barriers to access public health services, which might inhibit migrants from seeking those services. In order to comply with its commitment to guarantee the right to health of all persons, the Mexican health authorities should address the implementation gap between an inclusive policy, and the barriers to access that still remain.

Abstract In this work we use Mimosa tenuiflora (MtE) extracts as reducing agents to synthesize silver nanoparticles (AgMt NPs) which were characterized by DPPH and Total Polyphenols Assays, UV–visible, X-ray diffractometer (XRD), high-resolution transmission electron microscopy (HRTEM), X-ray photoelectron spectroscopy (XPS) and Thermogravimetric analysis (TGA). AgMt NPs possess average sizes of 21 nm and fcc crystalline structure, it was also confirmed that the MtE is present in the AgMt NPs even after the cleaning protocol applied. Subsequently, carbopol hydrogels were made and the MtE and the synthesized AgMt NPs were dispersed in different gels (MtE-G and AgMt NPs-G, respectively) at 100 µg/g concentration. The gels were characterized by UV–Vis, IR, and rheology. Antimicrobial tests were performed using Staphylococcus aureus and Escherichia coli. Burn wound healing was evaluated in a second-degree burn injury on a Wistar rats model for 14 days and additional skin biopsies were examined with histopathological analysis. Gel with commercial silver nanoparticles (Ag NPs) was prepared and employed as a control on the biological assays. Hydrogel system containing silver nanoparticles synthesized with Mimosa tenuiflora (AgMt NPs-G) is a promising therapeutic strategy for burn wound healing, this due to bactericidal and anti-inflammatory effects, which promotes a more effective recovery (in percentage terms) by damaged area.

Beyond sleep/wake, clock genes regulate the daily rhythms of melatonin production, motor activity, innate immunity, and mitochondrial dynamics, among others. All these rhythms are affected in Parkinson’s disease (PD), suggesting that chronodisruption may be an early stage of the disease. The aim of this study was to evaluate the connection between clock genes and these rhythms in PD, and whether melatonin administration reestablished the normal clock function. Parkinsonism was induced with 600 μM MPTP (N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) in 24–120 h post fertilization (hpf) zebrafish embryos and melatonin was administered at a dose of 1 μM. Day–night melatonin rhythm disappeared in MPTP-treated embryos, which showed an advance in the activity phase in parallel with changes in the rhythm of clock genes. An alteration in the fission-to-fusion mitochondrial dynamics was also detected in parkinsonian embryos, increasing the former and leading to apoptosis. Melatonin administration to MPTP-treated embryos fully restored the circadian system, including the rhythms of clock genes, motor activity, melatonin rhythm, and mitochondrial dynamics, and decreasing apoptosis. Because clock-controlled rhythms such as sleep/wake alterations are early events in PD, the data here reported may point to chronodisruption as one initial pathophysiological event of the disease.

El encuentro con la obra y pensamiento de Bertolt Brecht puede ser considerado un momento crucial para el desarrollo artístico del cantautor y poeta portugués José Afonso (Aveiro, 2 de agosto de 1929—Setúbal, 23 de febrero de 1987), dibujando un marco creativo que se ampliará a partir de 1967 como contexto teórico y práctico desde el que entender mucha de su producción. En este sentido, José Afonso lleva al terreno musical popular portugués la reivindicación de Brecht como modelo de intervención cultural que impregna la oposición socialista a la dictadura de Salazar. De este modo, es posible identificar tanto en sus ideas como en sus prácticas compositivas e interpretativas una apropiación consciente de las concepciones brechtianas. Esta apropiación se produce no solo en lo que concierne al papel cultural y político del músico y su música en la ruptura del monolítico orden cultural salazarista, sino también en una dimensión intrínsecamente estética, en la propia utilización de voces cronológica y estilísticamente múltiples para ensamblar artefactos de gran poder expresivo y movilizador. En las canciones de José Afonso, particularmente en las que compuso sobre textos de Brecht, están presentes muchos de los principios y procedimientos constructivos que sitúan al dramaturgo en una línea simultáneamente moderna y políticamente comprometida. Especialmente destacable es la transferencia a la música de los métodos de montaje y superposición de lenguajes, lo que se traduce en un juego ambivalente entre lo tradicional y lo urbano para promover una reconstrucción crítica de lo real. Esto permitiría definir su producción como moderna y popular.

The major toxins of Clostridioides difficile (TcdA, TcdB, CDT) are chromosomally encoded in nearly all known strains. Following up on previous findings, we identified 5 examples of a family of putative conjugative plasmids with tcdB and cdtAB in clinical C. difficile isolates from multilocus sequence typing clades C-I, 2, and 4.

Abstract Background Based on MLST analyses the global population of C. difficile is distributed in eight clades, of which Clade 2 includes the “hypervirulent” NAP1/RT027/ST01 strain along with various unexplored sequence types (STs). Methods To clarify whether this clinically relevant phenotype is a widespread feature of C. difficile Clade 2, we used the murine ileal loop model to compare the in vivo pro-inflammatory (TNF-α, IL-1β, IL-6) and oxidative stress activities (MPO) of five Clade 2 clinical C. difficile isolates from sequence types (STs) 01, 41, 67, and 252. Besides, we infected Golden Syrian hamsters with spores from these strains to determine their lethality, and obtain a histological evaluation of tissue damage, WBC counts, and serum injury biomarkers (LDH, ALT, AST, albumin, BUN, creatinine, Na+, and Cl−). Genomic distances were calculated using Mash and FastANI to explore whether the responses were dictated by phylogeny. Results The ST01 isolate tested ranked first in all assays, as it induced the highest overall levels of pro-inflammatory cytokines, MPO activity, epithelial damage, biochemical markers, and mortality measured in both animal models. Statistically indistinguishable or rather similar outputs were obtained for a ST67 isolate in tests such as tissue damage, neutrophils count, and lethal activity. The results recorded for the two ST41 isolates tested were of intermediate magnitude and the ST252 isolate displayed the lowest pathogenic potential in all animal experiments. This ordering matched the genomic distance of the ST01 isolate to the non-ST01 isolates. Conclusions Despite their close phylogenic relatedness, our results demonstrate differences in pathogenicity and virulence levels in Clade 2 C. difficile strains, confirm the high severity of infections caused by the NAP1/RT027/ST01 strain, and highlight the importance of C. difficile typing.

After twenty-five years of climate litigation dominated by cases in the United States, Australia, and other jurisdictions in the Global North, a second wave of lawsuits arose in the mid-2010s that prominently feature cases filed in countries of the Global South. I argue that the use of human rights norms and strategies characterizes the “Global South route” to climate litigation, one that is firmly rooted in the trajectory of human rights adjudication and litigation in key Southern countries over the last three decades. I posit that, in order to understand the present and the future of this route, it is essential to (1) track its origins and features to the trajectory of “Global South constitutionalism” over the last three decades, especially litigation around socioeconomic rights, and (2) unpack the category of “Global South” countries, in order to avoid overgeneralizations and to identify the types of countries that are likely to see most climate litigation and court decisions. I close by suggesting that, in light of the planetary and urgent nature of the climate challenge, future research and advocacy should explore transnational forms of litigation that cut across the North-South divide and pay systematic attention to the impact of climate litigation.

Clostridioides difficile infection (CDI) remains a significant healthcare burden. Non-toxigenic C. difficile (NTCD) strains have shown a benefit in preventing porcine enteritis and in human recurrent CDI. In this study, we evaluated the efficacy of metronidazole-resistant NTCD-E4 in preventing CDI facilitated by a range of antimicrobials in an in vitro human gut model. NTCD-E4 spores (at a dose of 107) were instilled 7 days before a clinical ribotype (RT) 027 (at the same dose) strain (210). In separate experiments, four different antimicrobials were used to perturb gut microbiotas; bacterial populations and cytotoxin production were determined using viable counting and Vero cell cytotoxicity, respectively. RT027 and NTCD-E4 proliferated in the in vitro model when inoculated singly, with RT027 demonstrating high-level cytotoxin (3-5-log10-relative units) production. In experiments where the gut model was pre-inoculated with NTCD-E4, RT027 was remained quiescent and failed to produce cytotoxins. NTCD-E4 showed mutations in hsmA and a gene homologous to CD196-1331, previously linked to medium-dependent metronidazole resistance, but lacked other metronidazole resistance determinants. This study showed that RT027 was unable to elicit simulated infection in the presence of NTCD-E4 following stimulation by four different antimicrobials. These data complement animal and clinical studies in suggesting NTCD offer prophylactic potential in the management of human CDI.

The circadian clock is a regulatory system, with a periodicity of approximately 24 h, that generates rhythmic changes in many physiological processes. Increasing evidence links chronodisruption with aberrant functionality in clock gene expression, resulting in multiple diseases, including cancer. In this context, tumor cells have an altered circadian machinery compared to normal cells, which deregulates the cell cycle, repair mechanisms, energy metabolism and other processes. Melatonin is the main hormone produced by the pineal gland, whose production and secretion oscillates in accordance with the light:dark cycle. In addition, melatonin regulates the expression of clock genes, including those in cancer cells, which could play a key role in the numerous oncostatic effects of this hormone. This review aims to describe and clarify the role of clock genes in cancer, as well as the possible mechanisms of the action of melatonin through which it regulates the expression of the tumor’s circadian machinery, in order to propose future anti-neoplastic clinical treatments.

La siguiente investigación aborda el estudio pormenorizado de un vestido oficial de la corte Qing también conocido como jifu. Perteneciente a la colección del Kunstmuseum Den Haag (KMDH) en los Países Bajos, dicho vestido ha contribuido a interpretar y contextualizar el momento en el que el comercio de objetos textiles de Oriente se encontraba en pleno auge en Occidente a través del movimiento cultural conocido como Orientalismo. Originalmente confeccionado para ser vestido por un hombre de la corte, la prenda refleja en la actualidad la característica forma de un abrigo de mujer de finales de los años 20. En el presente estudio, la aplicación de técnicas de análisis científico, a través de la colaboración del Instituto Valenciano de Conservación y Restauración e Investigación (IVCR+i), ayudó no solamente a determinar la calidad de los materiales presentes, sino también, a justificar los elementos de degradación activos en el momento de estudio del vestido. Todo ello, junto con el análisis de los cambios sufridos respecto al diseño original, ayudó a determinar el mejor tratamiento de conservación y restauración posible para salvaguardar tan preciado bien cultural.

Multilocus Sequence Typing (MLST) is a precise microbial typing approach at the intra-species level for epidemiologic and evolutionary purposes. It operates by assigning a sequence type (ST) identifier to each specimen, based on a combination of alleles of multiple housekeeping genes included in a defined scheme. The use of MLST has multiplied due to the availability of large numbers of genomic sequences and epidemiologic data in public repositories. However, data processing speed has become problematic due to the massive size of modern datasets. Here, we present FastMLST, a tool that is designed to perform PubMLST searches using BLASTn and a divide-and-conquer approach that processes each genome assembly in parallel. The output offered by FastMLST includes a table with the ST, allelic profile, and clonal complex or clade (when available), detected for a query, as well as a multi-FASTA file or a series of FASTA files with the concatenated or single allele sequences detected, respectively. FastMLST was validated with 91 different species, with a wide range of guanine-cytosine content (%GC), genome sizes, and fragmentation levels, and a speed test was performed on 3 datasets with varying genome sizes. Compared with other tools such as mlst, CGE/MLST, MLSTar, and PubMLST, FastMLST takes advantage of multiple processors to simultaneously type up to 28 000 genomes in less than 10 minutes, reducing processing times by at least 3-fold with 100% concordance to PubMLST, if contaminated genomes are excluded from the analysis. The source code, installation instructions, and documentation of FastMLST are available at https://github.com/EnzoAndree/FastMLST

The intestinal pathogen Clostridioides (C.) difficile is a major cause of diarrhea both in hospitals and outpatient in industrialized countries. This bacterium produces two large exotoxins, toxin A (TcdA) and toxin B (TcdB), which are directly responsible for the onset of clinical symptoms of C. difficile-associated diseases (CDADs), such as antibiotics-associated diarrhea and the severe, life-threatening pseudomembranous colitis. Both toxins are multidomain proteins and taken up into host eukaryotic cells via receptor-mediated endocytosis. Within the cell, TcdA and TcdB inactivate Rho and/or Ras protein family members by glucosylation, which eventually results in cell death. The cytotoxic mode of action of the toxins is the main reason for the disease. Thus, compounds capable of inhibiting the cellular uptake and/or mode-of-action of both toxins are of high therapeutic interest. Recently, we found that the sterol regulatory element-binding protein 2 (SREBP-2) pathway, which regulates cholesterol content in membranes, is crucial for the intoxication of cells by TcdA and TcdB. Furthermore, it has been shown that membrane cholesterol is required for TcdA- as well as TcdB-mediated pore formation in endosomal membranes, which is a key step during the translocation of the glucosyltransferase domain of both toxins from endocytic vesicles into the cytosol of host cells. In the current study, we demonstrate that intoxication by TcdA and TcdB is diminished in cultured cells preincubated with the compound U18666A, an established inhibitor of cholesterol biosynthesis and/or intracellular transport. U18666A-pretreated cells were also less sensitive against TcdA and TcdB variants from the epidemic NAP1/027 C. difficile strain. Our study corroborates the crucial role of membrane cholesterol for cell entry of TcdA and TcdB, thus providing a valuable basis for the development of novel antitoxin strategies in the context of CDADs.

Clostridioides difficile infection (CDI) remains an urgent global One Health threat. The genetic heterogeneity seen across C. difficile underscores its wide ecological versatility and has driven the significant changes in CDI epidemiology seen in the last 20 years. We analysed an international collection of over 12,000 C. difficile genomes spanning the eight currently defined phylogenetic clades. Through whole-genome average nucleotide identity, and pangenomic and Bayesian analyses, we identified major taxonomic incoherence with clear species boundaries for each of the recently described cryptic clades CI–III. The emergence of these three novel genomospecies predates clades C1–5 by millions of years, rewriting the global population structure of C. difficile specifically and taxonomy of the Peptostreptococcaceae in general. These genomospecies all show unique and highly divergent toxin gene architecture, advancing our understanding of the evolution of C. difficile and close relatives. Beyond the taxonomic ramifications, this work may impact the diagnosis of CDI.

Abstract Synthesis of gold nanoparticles (AuNPs) with plant extracts has gained great interest in the field of biomedicine due to its wide variety of health applications. In the present work, AuNPs were synthesized with Mimosa tenuiflora (Mt) bark extract at different metallic precursor concentrations. Mt extract was obtained by mixing the tree bark in ethanol-water. The antioxidant capacity of extract was evaluated using 2,2-diphenyl-1-picrylhydrazyl and total polyphenol assay. AuNPs were characterized by transmission electron microscopy, X-ray diffraction, UV-Vis and Fourier transform infrared spectroscopy, and X-ray photoelectron spectrometry for functional group determination onto their surface. AuMt (colloids formed by AuNPs and molecules of Mt) exhibit multiple shapes with sizes between 20 and 200 nm. AuMt were tested on methylene blue degradation in homogeneous catalysis adding sodium borohydride. The smallest NPs (AuMt1) have a degradation coefficient of 0.008/s and reach 50% degradation in 190s. Cell viability and cytotoxicity were evaluated in human umbilical vein endothelial cells (HUVEC), and a moderate cytotoxic effect at 24 and 48 h was found. However, toxicity does not behave in a dose-dependent manner. Cellular internalization of AuMt on HUVEC cells was analyzed by confocal laser scanning microscopy. For AuMt1, it can be observed that the material is dispersed into the cytoplasm, while in AuMt2, the material is concentrated in the nuclear periphery.

Abstract Background When present in animal feedstuff, mycotoxins contaminants and antibiotic residues can have negative implications for animal production and Public Health, including the transmission of carcinogenic compounds and the selection of antibiotic resistant bacteria, respectively. So far there are no available methods in which both mycotoxins and antibiotic residues are analyzed using a parallel extraction approach. To address this issue, we developed a LC/MS methodology with high sensitivity (0.005 to 6.42 and 24.55 to 132.73 μg kg− 1 for mycotoxins and antimicrobials) and specificity (unique target ion mass/charge) that allows the detection of 26 mycotoxins and 23 antibiotic residues in animal feedstuff and validated it through the determination of these analytes in 294 animal feed and feed ingredient samples in the framework of a country-wide surveillance program. Two hundred and five of these samples were analyzed for mycotoxins and 89 for antibiotics. Findings Fumonisin was the most frequently toxin found, with FB1 and FB2 presenting prevalences of 50 and 52% and maximum concentrations of 14,927.61 and 8646.67 μg kg− 1, respectively. Other toxins, including diacetoxyscirpenol n = 4/101 (3.96%), fusarenon-X n = 2/101 (1.98%), citrinin n = 2 (1.98%), and patulin n = 1 (0.99%) were rarely found. Toxicologically relevant concentrations of toxin metabolites, such as HT-2 (6.38–485.49 μg kg− 1) and 3−/15-acetoxydeoxynivalenol (877.89–3236.56/5.44–1685.3 μg kg− 1), were also found. Few samples exceeded threshold mycotoxin concentrations defined in current EU guidelines. Dairy cattle and swine feeds included the higher number of samples exceeding guideline values (n = 6 and n = 5, respectively). From the total of samples analysed for antibiotics, 7.7% (n = 7/89) were classified as medicated for poultry and pigs. Unexpectedly, 57% of these medicated samples contained no detectable antibiotics (n = 4/7). The remaining 43% of the samples (n = 3/7) presented inconsistencies regarding the concentration of analytes declared on the labels or the antibiotics found. Likewise 74.6% (n = 50/67) of the non-medicated feed samples analyzed had antibiotic residues. Additionally, we analyzed commercial monensin standards for purity and evaluate batch-to-batch flushing feed industry practices. Conclusions Herein we report the results for a year-wide analysis for mycotoxins and antibiotics in feed samples. Mycotoxins, several metabolites, and the occurrence of these emerging contaminants were evaluated and antibiotic residues in non-medicated feed samples were found using a targeted MS-based LC approach. This validated multi-analyte method is expected to facilitate the monitoring and surveillance of contaminants, from natural and anthropogenic origin, in animal feed.

The zebrafish has become an excellent model for the study of human diseases because it offers many advantages over other vertebrate animal models. The pineal gland, as well as the biological clock and circadian rhythms, are highly conserved in zebrafish, and melatonin is produced in the pineal gland and in most organs and tissues of the body. Zebrafish have several copies of the clock genes and of aanat and asmt genes, the latter involved in melatonin synthesis. As in mammals, melatonin can act through its membrane receptors, as with zebrafish, and through mechanisms that are independent of receptors. Pineal melatonin regulates peripheral clocks and the circadian rhythms of the body, such as the sleep/wake rhythm, among others. Extrapineal melatonin functions include antioxidant activity, inducing the endogenous antioxidants enzymes, scavenging activity, removing free radicals, anti-inflammatory activity through the regulation of the NF-κB/NLRP3 inflammasome pathway, and a homeostatic role in mitochondria. In this review, we introduce the utility of zebrafish to analyze the mechanisms of action of melatonin. The data here presented showed that the zebrafish is a useful model to study human diseases and that melatonin exerts beneficial effects on many pathophysiological processes involved in these diseases.

Los programas de seguridad basados en el comportamiento se aplican con la finalidad de reforzar el nivel de seguridad en el trabajo, pero si se hace por cumplir con un número de observaciones de comportamiento seguro establecidas en la meta del mes en el lugar de trabajo, genera en los trabajadores que solo cumplan con la seguridad mientras se les está observando o que los observadores reporten comportamientos que realmente no vieron; en consecuencia, los accidentes con trabajadores continúan y peor aún con casos fatales, los observadores dejan de creer en el programa de seguridad que están utilizando y la dirección llega a creer que con un mayor número de observaciones se logra corregir los comportamientos inseguros; por ello es que los resultados del programa no se alcanzan con el tiempo. Esto es consecuencia de no haber entendido los principios de la teoría de la seguridad basada en el comportamiento desde el inicio de su implementación. En la presente investigación se aplicó la teoría de la seguridad basada en los comportamientos para definir la tarjeta de observadores y para realizar las intervenciones con refuerzo positivo de manera eficiente. Así, el estudio se orientó a encontrar si hay diferencia significativa entre las observaciones de comportamiento seguro antes que el observador realice la retroalimentación con refuerzo positivo y las observaciones después de cada intervención, para ello se tomó sólo el número de observaciones seguras con el propósito de conocer el efecto que tiene la retroalimentación con refuerzo positivo que se da en la observación de comportamientos seguros en el periodo de estudio. Se encontró que hay diferencia significativa en los comportamientos seguros después de realizar la retroalimentación para un nivel de confianza del 95% y una reducción del 85.7% de accidentes incapacitantes en el periodo de estudio.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease. Lesions in the lung epithelium cause alterations in the microenvironment that promote fibroblast accumulation. Extracellular vesicles (EVs) transport proteins, lipids, and nucleic acids, such as microRNAs (miRNAs). The aim of this study was to characterize the differentially expressed miRNAs in the cargo of EVs obtained from the LL97 and LL29 fibroblast cell lines isolated from IPF lungs versus those derived from the CCD19 fibroblast cell line isolated from a healthy donors. We characterized EVs by ultracentrifugation, Western blotting, and dynamic light scattering. We identified miRNAs by small RNA-seq, a total of 1144 miRNAs, of which 1027 were known miRNAs; interestingly, 117 miRNAs were novel. Differential expression analysis showed that 77 miRNAs were upregulated and 68 were downregulated. In addition, pathway enrichment analyses from the Gene Ontology and Kyoto Encyclopedia of Genomes identified several miRNA target genes in the categories, cell proliferation, regulation of apoptosis, pathways in cancer, and proteoglycans in cancer. Our data reveal that miRNAs contained in EVs cargo could be helpful as biomarkers for fibrogenesis, diagnosis, and therapeutic intervention of IPF.

Introduction. The coffee berry borer (Hypothenemus hampei Ferrari, CBB) is one of the most devastating pests on coffee plantations around the world. Although CBB is susceptible to the effect of δ–endotoxins of Bacillus thuringiensis subs. israelensis (Bti) at laboratory level, the efficacy of this control method is poor in the field, presumably due to the inactivation by digestive proteases different to those required for protoxin activation. Objective. To study whether the addition of a soybean flour extract enriched with protease inhibitors (PI), mixed with Bti crystals and spores (Bti-sc) in an artificial diet, could improve the toxicity of Bti against CBB. Materials and methods. This study was performed in San José, Costa Rica, between 2012 and 2013. A set of adult female CBB insects was exposed to a mixture containing different concentrations of a partially purified soybean meal extract with active PI and lyophilized Bti-sc, and were tested through a bioassay in artificial diet to estimate the sub-lethal concentration (LC50). The mortality results were validated by observing the dissected midgut, whose ultrastructure was analyzed by transmission electron microscopy. Results. The soybean extracts partially degraded the Bti-sc complex, it reduced its LC50 by almost four times (from 1.135 to 0.315 µg µl-1) and enhanced CBB mortality in a concentration-dependent manner. Histological analyses of the midgut confirmed this synergistic effect, since severe epithelial damage to the intestinal epithelium of CBB exposed to Bti-sc + PI was visualized compared to Bti-sc alone. Conclusions. The combination of a soybean extract enriched in PI and Bti-sc enhanced the mortality effect over CBB, which was confirmed by the midgut collapse. Soybean flour is a cost-effective supplement that could increase Bti effectiveness against CBB and delay the appearance of biological resistance.

Here, we present the characterization of three iPSC lines derived from dermal fibroblasts of old healthy subjects. Fibroblasts were reprogrammed using Sendai viral vectors encoding OCT4, SOX2, KLF4 and c-MYC. The iPSCs expressed endogenous pluripotency markers, could generate the three germ layers (ectoderm, mesoderm and endoderm), maintained a stable karyotype, and were free from Sendai vectors and reprogramming factors. These integration-free iPSCs can serve for establishing control cell cultures in studies searching for phenotypes and mechanisms that could potentially be dysregulated in degenerative diseases.

A reliable estimate of the wind potential in the marine atmospheric boundary layer (MABL) is of great importance to justify the energetic viability of new offshore wind farms. The purpose of the study is to provide an additional tool for the prediction of the energy that a wind turbine would produces in the open sea from the usual way of measurements at sea, that is, when they are carried out with measuring masts, where the meteorological data are obtained at levels much lower than those of a wind turbine hub. For this, the variation in the wind speed with the height in the MABL is determined, based on the Monin–Obukhov similarity theory, according to the boundary conditions of the air–sea interface, where the input data for the Validation of the results are extracted from the German FINO 3 research platform during the years 2016, 2017, and 2018. It is applied to the production of electrical energy from a 6.0 MW commercial wind turbine, with the hub at 100 m above the sea surface. As a more prominent result, the deviations from the proposed method do not exceed 2.5% in the energy calculation.

The epsilon4 (ε4) allele of the APOE gene, which encodes the apolipoprotein E4 (ApoE4), is the strongest genetic risk factor known for late-onset Alzheimer´s disease (LOAD). Here, we present the characterization of an iPSC line generated from dermal fibroblasts of a female AD patient using Sendai viral vectors encoding the transcription factors OCT4, SOX2, KLF4 and c-MYC. The iPSCs maintained the original genotype, a normal karyotype, were free from Sendai viral vectors and reprogramming factors, presented a normal morphology, expressed endogenous pluripotency markers, and could be differentiated into ectodermal, mesodermal and endodermal cells, confirming its pluripotency.

En el artículo se expone la necesidad de la gestión neuropedagógica de los procesos de aprendizaje, considerándose como problema de investigación la reorientación de las prácticas pedagógicas del docente con elementos neuropedagógicos. La inadecuada aplicación de estrategias de enseñanza, la falta de preparación neuropedagógica de los docentes, y la omisión de recursos para gestionar la motivación intrínseca hacia el aprendizaje, genera conformismo y desinterés en los estudiantes restándole significatividad al aprendizaje. Como consecuencia resulta novedoso en el contexto ecuatoriano profundizar desde la investigación pedagógica en lo referido a la optimización de los procesos de aprendizaje con el apoyo de técnicas de superaprendizaje enfocadas hacia el desarrollo neurobiopsicosocial de los estudiantes, y la evaluación de su impacto en el desarrollo integral del estudiante. ABSTRACT In the article, the need of the neuropedagogical management of the learning processes is exposed, considering as a research problem the reorientation of the pedagogical practices of the teacher with neuropedagogical elements. The inadequate application of teaching strategies, the lack of neuropedagogical preparation of teachers, and the omission of resources to manage the intrinsic motivation towards learning, generates conformism and disinterest in students, reducing significance to learning. As a result, it is new in the Ecuadorian context to deepen from the pedagogical research regarding the optimization of the learning processes with the support of superlearning techniques focused on the neurobio-psychosocial development of the students, and the evaluation of their impact on the integral development of student. KEY WORDS: Learning, Brain, Integral Development, Superlearning Techniques, Neuropedagogy.

El Servicio Geográfico Militar (SGM) de Uruguay es la historia misma de los hom- bres y mujeres que lo integraron. Sacrificio, conocimiento y disciplina, jalonan los primeros 100 años. Su herramienta fundamental para la mejora continua, ha sido su gente, su férrea voluntad de hacer y de sobreponerse a los obstáculos. Ellos, genera- ron el legado, la tradición, a las que se suman las diferentes circunstancias y decisiones que debieron asumir, sus fortalezas y debilidades, sus aciertos y fallos,... los fallos que a todos, nos hace humanos.

The familial form of Alzheimer's disease (FAD), which is caused by mutations in PRESENILIN 1 (PSEN1) and amyloid precursor protein (APP) genes, represents less than 5% of all AD cases and has an early-onset. We report the generation and characterization of an iPSC line derived from a FAD patient carrying the PSEN1-G206D mutation. The iPSC line maintained the original genotype, a normal karyotype, was free from Sendai viral vectors and reprogramming factors (OCT4, SOX2, KLF4 and c-MYC), presented a typical morphology, expressed endogenous pluripotency markers, and could be differentiated into ectodermal, mesodermal and endodermal cells, confirming its pluripotency.

The L444P mutation in the GBA1 gene which encodes β-glucocerebrosidase-1, is a major risk factor for developing Parkinson's disease (PD) and dementia with Lewy bodies (DLB). We report the generation and characterization of an induced pluripotent stem cell (iPSC) line derived from a female PD patient carrying the L444P/wt mutation. The iPSC line presented a normal morphology, expressed endogenous pluripotency markers, could be differentiated into endodermal, mesodermal and ectodermal cells, was free from Sendai vectors and reprogramming factors, had a normal karyotype and maintained the original GBA1 genotype. Thus, this iPSC line can serve to establish cellular models of PD.

Genetic polymorphism of apolipoprotein E (APOE) confers differential susceptibility to late-onset Alzheimer's disease (LOAD). The ε3 allele of APOE, the most common isoform, does not represent a risk factor for LOAD. In contrast, the ε4 allele is the strongest genetic risk factor for this disease. Here, we present the characterization of four iPSC lines generated from dermal fibroblasts of diagnosed sporadic AD patients using Sendai viral vectors encoding OCT4, SOX2, KLF4 and c-MYC. The iPSCs expressed endogenous pluripotency markers, could be differentiated into the three germ layers, maintained the original genotypes, and were free from Sendai vectors and reprogramming factors.

Mutations in the GBA1 gene, which encodes the lysosomal enzyme Glucocerebrosidase1 are major risk factors for Parkinson's disease (PD) and dementia with Lewy bodies (DLB). We have generated induced pluripotent stem cells (iPSCs) from fibroblasts of four PD patients carrying the N370S/wt or the L444P/wt heterozygous mutations in GBA1. The iPSCs presented a normal morphology, expressed endogenous pluripotency markers and could be differentiated into endodermal, mesodermal and ectodermal cells. These iPSCs were free from Sendai viral vectors and reprogramming factors, had a normal karyotype and maintained the original GBA1 genotype.

This dissertation attends to the study of football hooligans’ subcultures. In particular, it addresses a general synthesis of the beginnings of casual culture in Great Britain, within the context of the cultural transition process of the 1980s, and within a political, social and cultural context greatly influenced by the new Conservative government of Margaret Thatcher. It makes a chronological review of the stylistic evolution and the attitudes of the casuals, based on the concept one-upmanship, facing the different realities that happened in approximately a decade. From the birth of the punk movement in the late seventies to the emergence of rave and club cultures at the end of the following decade. It also includes the element of violence in football, both inside and outside the stadiums, through several events that exemplify the level of violence achieved in those years. Throughout the text it tries to record the relevance of the study of youth expressions and activities for a better understanding of wider historical and cultural processes.

Metabolic reprogramming, which is characteristic of cancer cells that rapidly adapt to the hypoxic microenvironment and is crucial for tumor growth and metastasis, is recognized as one of the major mechanisms underlying therapeutic resistance. Mitochondria, which are directly involved in metabolic reprogramming, are used to design novel mitochondria-targeted anticancer agents. Despite being targeted by melatonin, the functional role of mitochondria in melatonin’s oncostatic activity remains unclear. In this study, we aim to investigate the role of melatonin in mitochondrial metabolism and its functional consequences in head and neck cancer. We analyzed the effects of melatonin on head and neck squamous cell carcinoma (HNSCC) cell lines (Cal-27 and SCC-9), which were treated with 100, 500, and 1500 µM of melatonin for 1, 3, and 5 days, and found a connection between a change of metabolism following melatonin treatment and its effects on mitochondria. Our results demonstrate that melatonin induces a shift to an aerobic mitochondrial metabolism that is associated with changes in mitochondrial morphology, function, fusion, and fission in HNSCC. We found that melatonin increases oxidative phosphorylation (OXPHOS) and inhibits glycolysis in HNSCC, resulting in increased ROS production, apoptosis, and mitophagy, and decreased cell proliferation. Our findings highlight new molecular pathways involved in melatonin’s oncostatic activity, suggesting that it could act as an adjuvant agent in a potential therapy for cancer patients. We also found that high doses of melatonin, such as those used in this study for its cytotoxic impact on HNSCC cells, might lead to additional effects through melatonin receptors.

Muscular aging is a complex process and underlying physiological mechanisms are not fully clear. In recent years, the participation of the NF-kB pathway and the NLRP3 inflammasome in the chronic inflammation process that accompanies the skeletal muscle’s aging has been confirmed. microRNAs (miRs) form part of a gene regulatory machinery, and they control numerous biological processes including inflammatory pathways. In this work, we studied the expression of four miRs; three of them are considered as inflammatory-related miRs (miR-21, miR-146a, and miR-223), and miR-483, which is related to the regulation of melatonin synthesis, among other targets. To investigate the changes of miRs expression in muscle along aging, the impact of inflammation, and the role of melatonin in aged skeletal muscle, we used the gastrocnemius muscle of wild type (WT) and NLRP3-knockout (NLRP3−) mice of 3, 12, and 24 months-old, with and without melatonin supplementation. The expression of miRs and pro-caspase-1, caspase-3, pro-IL-1β, bax, bcl-2, and p53, was investigated by qRT-PCR analysis. Histological examination of the gastrocnemius muscle was also done. The results showed that age increased the expression of miR-21 (p < 0.01), miR-146a, and miR-223 (p < 0.05, for both miRs) in WT mice, whereas the 24-months-old mutant mice revealed decline of miR-21 and miR-223 (p < 0.05), compared to WT age. The lack of NLRP3 inflammasome also improved the skeletal muscle fibers arrangement and reduced the collagen deposits compared with WT muscle during aging. For the first time, we showed that melatonin significantly reduced the expression of miR-21, miR-146a, and miR-223 (p < 0.05 for all ones, and p < 0.01 for miR-21 at 24 months old) in aged WT mice, increased miR-223 in NLRP3− mice (p < 0.05), and induced miR-483 expression in both mice strains, this increase being significant at 24 months of age.

Clostridium difficile is the major causative agent of nosocomial antibiotic-associated diarrhea. In a 2009 outbreak of C. difficile-associated diarrhea that was recorded in a major Costa Rican hospital, the hypervirulent NAP1 strain (45%) predominated together with a local genotype variant (NAPCR1, 31%). Both strains were fluoroquinolone-resistant and the NAPCR1 genotype, in addition, was resistant to clindamycin and rifampicin. We now report on the genotypes and antibiotic susceptibilities of 68 C. difficile isolates from a major Costa Rican hospital over a 2-year period without outbreaks. In contrast to our previous findings, no NAP1 strains were detected, and for the first time in a Costa Rican hospital, a significant fraction of the isolates were NAP9 strains (n=14, 21%). The local NAPCR1 genotype remained prevalent (n=18, 26%) and coexisted with 14 strains (21%) of classic hospital NAP types (NAP2, NAP4, and NAP6), eight new genotypes (12%), four environmental strains classified as NAP10 or NAP11 (6%), three strains without NAP designation (4%) and seven non-toxigenic strains (10%). All 68 strains were resistant to ciprofloxacin, 88% were resistant to clindamycin and 50% were resistant to moxifloxacin and rifampicin. Metronidazole and vancomycin susceptibilities were universal. The NAPCR1 and NAP9 strains, which have been associated with more severe clinical infections, were more resistant to antibiotics than the other strains. Altogether, our results confirm that the epidemiology of C. difficile infection is dynamic and that A−B+ strains from the NAP9 type are on the rise not only in the developed world. Moreover, our results reveal that the local NAPCR1 strains still circulate in the country without causing outbreaks but with equally high antibiotic-resistance rates and levels.

Emotional intelligence (EI) and mindfulness are two constructs that have been separately studied, and the relation between them still remains unclear. Research in this area has not attempted to go further into how enhancing EI and mindfulness together can achieve better improvements in this ability to attend mindfully. To bridge this knowledge gap, our research goal was to study the relationship between EI and the mindfulness competence in our study sample and to assess the impact of implementing EI and a mindfulness competence developmental program (SEA) about participants’ mindfulness competence. The sample consisted of 156 students aged 11–14 years old from a Spanish public high school. One hundred and eight participants were randomly assigned to the experimental condition, and the remaining 48 were to the control condition. The instruments used to evaluate EI were the CDE-SEC, EQi-Youth Version and the General Empathy Scale. Mindfulness on the School Scope Scale was used to assess mindfulness competences. Social adaptation was evaluated by using the social abilities and adjustment questionnaire BAS3. All the instruments where answered by the participants and have been adapted to a sample of youths with such age specifications. The results showed that EI and mindfulness were related to many of the variables measured by the instruments. Showing a good mindfulness competence was particularly related to having a good general level of the EI trait, and to many of the assessed social and emotional variables. The data indicated a significant relation between the mindfulness competence and having better general empathy skills or being better socially adjusted to the school context. The data also indicated a significant effect on participants’ interior and kinesthetic mindfulness competence after implementing the SEA Program. These findings corroborate the relationship between EI and mindfulness, and the possibility of enhancing mindfulness by applying a direct intervention program in the classroom.