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BackgroundIntegrating pediatric palliative care (PPC) into pediatric oncology standard care is essential. Therefore, it is important to assess physicians’ knowledge and perceptions of PPC to optimize its practice.ObjectiveTo evaluate the knowledge, comfort levels, and perspectives of physicians regarding the timing and perceived barriers to integrating PPC into pediatric cancer care across Europe.DesignThe Assessing Doctors’ Attitudes on Palliative Treatment (ADAPT) survey, originally developed for other global regions, was culturally and contextually adapted for Europe.Setting/SubjectsThe survey was distributed via the European Society of Paediatric Oncology (SIOPE) membership listserv. Any physicians caring for children with cancer across Eastern, Southern, Central, and Northern Europe were invited to complete the survey.ResultsA total of 198 physicians from 29 European countries completed the ADAPT survey. Physicians demonstrated relative agreement with the World Health Organization’s guidance; median alignment was 83.4% (range 59.9%-94.1%). Although most respondents felt comfortable addressing physical (84.4%) and emotional (63.4%) needs, they felt less comfortable addressing spiritual needs (41.9%) and providing grief and bereavement support (48.5%). There were significant regional differences, such as physicians in Eastern and Southern Europe reporting a lack of PPC specialists, opioids, and home-based care, while those in Northern and Central Europe did not.ConclusionPhysicians caring for children with cancer throughout Europe have a good understanding of PPC. However, misconceptions about PPC persist, requiring educational and capacity-building efforts. Additionally, the regional differences in perceived barriers must be addressed to ensure equitable access to PPC for all European children with cancer.

ABSTRACT While genome-wide transposon mutagenesis screens have identified numerous essential genes in the significant human pathogen Streptococcus pyogenes (group A Streptococcus or GAS), many of their functions remain elusive. This knowledge gap is attributed in part to the limited molecular toolbox for controlling GAS gene expression and the bacterium’s poor genetic transformability. CRISPR interference (CRISPRi), using catalytically inactive GAS Cas9 (dCas9), is a powerful approach to specifically repress gene expression in both bacteria and eukaryotes, but ironically, it has never been harnessed for controlled gene expression in GAS. In this study, we present a highly transformable and fully virulent serotype M1T1 GAS strain and introduce a doxycycline-inducible CRISPRi system for efficient repression of bacterial gene expression. We demonstrate highly efficient, oligo-based single guide RNA cloning directly to GAS, enabling the construction of a gene knockdown strain in just 2 days, in contrast to the several weeks typically required. The system is shown to be titratable and functional both in vitro and in vivo using a murine model of GAS infection. Furthermore, we provide direct in vivo evidence that the expression of the conserved cell division gene ftsZ is essential for GAS virulence, highlighting its promise as a target for emerging FtsZ inhibitors. Finally, we introduce SpyBrowse (https://veeninglab.com/SpyBrowse), a comprehensive and user-friendly online resource for visually inspecting and exploring GAS genetic features. The tools and methodologies described in this work are poised to facilitate fundamental research in GAS, contribute to vaccine development, and aid in the discovery of antibiotic targets.IMPORTANCEWhile group A Streptococcus (GAS) remains a predominant cause of bacterial infections worldwide, there are limited genetic tools available to study its basic cell biology. Here, we bridge this gap by creating a highly transformable, fully virulent M1T1 GAS strain. In addition, we established a tight and titratable doxycycline-inducible system and developed CRISPR interference (CRISPRi) for controlled gene expression in GAS. We show that CRISPRi is functional in vivo in a mouse infection model. Additionally, we present SpyBrowse, an intuitive and accessible genome browser (https://veeninglab.com/SpyBrowse). Overall, this work overcomes significant technical challenges of working with GAS and, together with SpyBrowse, represents a valuable resource for researchers in the GAS field.

One of the major challenges in gene therapy is the efficient and safe introduction of nucleic acids into eukaryotic cells. This process requires overcoming various biological barriers and navigating complex pathways to reach target cells and achieve their biological function. To address this obstacle, numerous transfection methods have been developed, including physical techniques and the use of genetic vectors, both viral and non-viral. However, to date, no transfection method is 100% safe and efficient. Within the spectrum of non-viral genetic vectors, cationic liposomes formed by cationic lipids stand out for their ability to protect and deliver therapeutic NA. These liposomes offer greater biocompatibility and lower immunogenicity compared to viral vectors, although they still do not match the efficiency of viral delivery systems. Consequently, ongoing research focuses on synthesizing a wide variety of cationic lipids in the search for compounds that provide high transfection efficiency with minimal cytotoxicity. This study aimed to design and synthesize a novel cationic lipid (CholCadLys) derived from natural cellular molecules for transferring genetic material to eukaryotic cells. The lipid was synthesized using cholesteryl chloroformate for the hydrophobic region, cadaverine as a linker, and lysine for the polar region, connected by carbamate and amide bonds, respectively. Identification was confirmed through thin-layer chromatography, purification through preparative chromatography, and characterization via infrared spectroscopy and mass spectrometry. The synthesis yielded a 60% success rate, with stable nanoliposomes averaging 76 nm in diameter. Liposomes were formed using this CL and commercial neutral lipids, characterized by transmission electron microscopy and Nanoparticle Tracking Analysis. These liposomes, combined with plasmid DNA, formed lipoplexes used to transfect Hek-293 FT cells, achieving up to 40% transfection efficiency without cytotoxicity in the mixture of CholCadLys and CholCad. This novel CL demonstrates potential as an efficient, safe, and cost-effective gene transfer system, facilitating further development in gene therapy.

Abstract The gene dosage compensation hypothesis presents a mechanism through which the expression of certain genes is modulated to compensate for differences in the dose of genes when additional chromosomes are present. It is one of the means through which cancer cells actively cope with the potential damaging effects of aneuploidy, a hallmark of most cancers. Dosage compensation arises through several processes, including downregulation or overexpression of specific genes and the relocation of dosage‐sensitive genes. In cancer, a majority of compensated genes are generally thought to be regulated at the translational or post‐translational level, and include the basic components of a compensation loop, including sensors of gene dosage and modulators of gene expression. Post‐translational regulation is mostly undertaken by a general degradation or aggregation of remaining protein subunits of macromolecular complexes. An increasingly important role has also been observed for transcriptional level regulation. This article reviews the process of targeted gene dosage compensation in cancer and other biological conditions, along with the mechanisms by which cells regulate specific genes to restore cellular homeostasis. These mechanisms represent potential targets for the inhibition of dosage compensation of specific genes in aneuploid cancers. This article critically examines the process of targeted gene dosage compensation in cancer and other biological contexts, alongside the criteria for identifying genes subject to dosage compensation and the intricate mechanisms by which cells orchestrate the regulation of specific genes to reinstate cellular homeostasis. Ultimately, our aim is to gain a comprehensive understanding of the intricate nature of a systems‐level property. This property hinges upon the kinetic parameters of regulatory motifs, which we have termed “gene dosage sensor loops.” These loops have the potential to operate at both the transcriptional and translational levels, thus emerging as promising candidates for the inhibition of dosage compensation in specific genes. Additionally, they represent novel and highly specific therapeutic targets in the context of aneuploid cancer.

Atherosclerosis is the leading cause of cardiovascular diseases in Mexico and worldwide. The membrane transporters ABCA1 and ABCG1 are involved in the reverse transport of cholesterol and stimulate the HDL synthesis in hepatocytes, therefore the deficiency of these transporters promotes the acceleration of atherosclerosis. MicroRNA-33 (miR-33) plays an important role in lipid metabolism and exerts a negative regulation on the transporters ABCA1 and ABCG1. It is known that by inhibiting the function of miR-33 with antisense RNA, HDL levels increase and atherogenic risk decreases. Therefore, in this work, a genetic construct, pPEPCK-antimiR-33-IRES2-EGFP, containing a specific antimiR-33 sponge with two binding sites for miR-33 governed under the PEPCK promoter was designed, constructed, and characterized, the identity of which was confirmed by enzymatic restriction, PCR, and sequencing. Hep G2 and Hek 293 FT cell lines, as well as a mouse hepatocyte primary cell culture were transfected with this plasmid construction showing expression specificity of the PEPCK promoter in hepatic cells. An analysis of the relative expression of miR-33 target messengers showed that the antimiR-33 sponge indirectly induces the expression of its target messengers (ABCA1 and ABCG1). This strategy could open new specific therapeutic options for hypercholesterolemia and atherosclerosis, by blocking the miR-33 specifically in hepatocytes.

This study investigated the changes that occurred during the COVID-19 pandemic in urban water consumption in residential, commercial, industrial, and public agencies in the city of São Leopoldo, southern Brazil, which has about 55,000 consumers and over 200,000 inhabitants. Overall, the city increased water consumption by 5.6% during the 2-year pandemic, with 5.9% in 2020 and 5.5% in 2021. Residential and industrial consumption increased by 6.77 and 9.92% in the first year, and by 5.47 and 14.45% in the second year, respectively. On the other hand, commercial and public sector consumption decreased by 5.48 and 46.26% in the first year and 1.83 and 40.99% in the second year, respectively. In the first months of the pandemic, there was a sharp increase in residential water consumption at the same time as a reduction in consumption in the other categories. In contrast, there was a slight return to previous water consumption patterns in the following months. Overall, we can affirm that the more central neighborhoods presented higher changes in water consumption than the peripheral neighborhoods. In addition, the water consumption during the pandemic and pre-pandemic periods was statistically different for residential, industrial, and public consumers. HIGHLIGHTS Residential water consumption has increased by 1 m³ per household per month.; Commercial and public establishments’ water consumption decreased sharply during two pandemic years.; The increase in COVID-19 containment measures generally causes an increase in residential water consumption.; After 2 years, there was a slight return to previous water consumption patterns.;

Helicobacter pylori have coevolved with mankind since its origins, adapting to different human groups. In America, H. pylori has evolved into several subpopulations. We analysed the genome of 154 Colombian strains along with 1,091 strains from worldwide populations to discern the ancestry and adaption to Colombian people. Population structure and ancestry was inferred with FineStructure and ChromoPainter. Phylogenetic relationship and the relative effect of recombination were analysing the core SNPs. Also, a Fst index was calculated to identify the gene variants with the strongest fixation in the Colombian subpopulations compared to their parent population hspSWEurope. FineStructure allowed the identification of two Colombian subpopulations, the previously described hspSWEuropeColombia and a novel subpopulation named hspColombia, that included three subgroups following their geographic origin. Colombian subpopulations represent an admixture of European, African and Indigenous ancestry; although some genomes showed a high proportion of self identity, suggesting an advanced adaption to these mestizo Colombian groups. We found that recombination is more important that punctual mutations in H. pylori genome diversity, 13.9 more important in hspSWEurope, 12.5 in hspSWEColombia and 10.5 in hspColombia, reflecting the divergence of these subpopulations. Fst analysis identified 82 SNPs fixed in 26 genes of the hspColombia subpopulation that encode for outer membrane and central metabolism proteins. Strongest fixation indexes were identified in genes encoding HofC, HopE, FrpB-4 and Sialidase A. These findings demonstrate that H. pylori has evolved in Colombia to give rise to subpopulations with a self identity ancestry, reflected in allele changes on genes encoding for outer membrane proteins.

Understanding a crop’s productive potential is crucial for optimizing resource use in agriculture, encouraging sustainable practices, and effectively planning planting and preservation efforts. Achieving precise and tailored management strategies is equally important. However, this task is particularly challenging in coffee cultivation due to the absence of accurate productivity maps for this crop. In this article, we created a multicriteria model to estimate the productive potential of coffee trees based on the observation of landscape elements that determine environmental fragility (EF). The model input parameters were slope and terrain shape data, slope flow power, and orbital image data (Landsat 8), allowing us to calculate the NDVI vegetation index. We applied the model developed to coffee trees planted in Bambuí, Minas Gerais, Brazil. We used seven plots to which we had access to yield data in a recent historical series. We compared the productivity levels predicted by the EF model and the historical productivity data of the coffee areas for the years 2016, 2018, and 2020. The model showed a high correlation between the calculated potential and the annual productivity. We noticed a strong correlation (R2) in the regression analyses conducted between the predicted productive potential and the actual productivity in 2018 and 2020 (0.91 and 0.93, respectively), although the correlation was somewhat weaker in 2016 (0.85). We conclude that our model could satisfactorily estimate the yearly production potential under a zero-harvest system in the study area.

IntroductionInborn errors of immunity (IEI) are an expanding group of rare diseases whose field has been boosted by next-generation sequencing (NGS), revealing several new entities, accelerating routine diagnoses, expanding the number of atypical presentations and generating uncertainties regarding the pathogenic relevance of several novel variants.MethodsResearch laboratories that diagnose and provide support for IEI require accurate, reproducible and sustainable phenotypic, cellular and molecular functional assays to explore the pathogenic consequences of human leukocyte gene variants and contribute to their assessment. We have implemented a set of advanced flow cytometry-based assays to better dissect human B-cell biology in a translational research laboratory. We illustrate the utility of these techniques for the in-depth characterization of a novel (c.1685G>A, p.R562Q) de novo gene variant predicted as probably pathogenic but with no previous insights into the protein and cellular effects, located in the tyrosine kinase domain of the Bruton’s tyrosine kinase (BTK) gene, in an apparently healthy 14-year-old male patient referred to our clinic for an incidental finding of low immunoglobulin (Ig) M levels with no history of recurrent infections.Results and discussionA phenotypic analysis of bone marrow (BM) revealed a slightly high percentage of pre-B-I subset in BM, with no blockage at this stage, as typically observed in classical X-linked agammaglobulinemia (XLA) patients. The phenotypic analysis in peripheral blood also revealed reduced absolute numbers of B cells, all pre-germinal center maturation stages, together with reduced but detectable numbers of different memory and plasma cell isotypes. The R562Q variant allows Btk expression and normal activation of anti-IgM-induced phosphorylation of Y551 but diminished autophosphorylation at Y223 after anti IgM and CXCL12 stimulation. Lastly, we explored the potential impact of the variant protein for downstream Btk signaling in B cells. Within the canonical nuclear factor kappa B (NF-κB) activation pathway, normal IκBα degradation occurs after CD40L stimulation in patient and control cells. In contrast, disturbed IκBα degradation and reduced calcium ion (Ca2+) influx occurs on anti-IgM stimulation in the patient’s B cells, suggesting an enzymatic impairment of the mutated tyrosine kinase domain.

Summary: Background: Data on risk factors for carbapenem-resistant Enterobacterales (CRE) with wider applicability are needed to inform preventive measures and efficient design of randomised trials. Methods: An international matched case-control-control study was performed in 50 hospitals with high CRE incidence from March 2016 to November 2018 to investigate different aspects of infections caused by CRE (NCT02709408). Cases were patients with complicated urinary tract infection (cUTI), complicated intraabdominal (cIAI), pneumonia or bacteraemia from other sources (BSI-OS) due to CRE; control groups were patients with infection caused by carbapenem-susceptible Enterobacterales (CSE), and by non-infected patients, respectively. Matching criteria included type of infection for CSE group, ward and duration of hospital admission. Conditional logistic regression was used to identify risk factors. Findings: Overall, 235 CRE case patients, 235 CSE controls and 705 non-infected controls were included. The CRE infections were cUTI (133, 56.7%), pneumonia (44, 18.7%), cIAI and BSI-OS (29, 12.3% each). Carbapenemase genes were found in 228 isolates: OXA-48/like, 112 (47.6%), KPC, 84 (35.7%), and metallo-β-lactamases, 44 (18.7%); 13 produced two. The risk factors for CRE infection in both type of controls were (adjusted OR for CSE controls; 95% CI; p value) previous colonisation/infection by CRE (6.94; 2.74–15.53;

The implementation of rainwater harvesting (RWH) systems depends on technical and socioeconomic assessments. However, most studies do not consider socioeconomic aspects, which could lead to different degrees of RWH implementation and technology selection due to economic constraints and local regulations. We evaluated the socioeconomic potential for RWH as an alternative for the water supply of 24 southern Brazilian municipalities with less than 50,000 inhabitants. A total of 10,080 RWH configurations were assessed and a reliability analysis was carried out to define the RWH system configurations potentially implementable (RWH+) in each municipality. RWH economic benefits were estimated from a social point of view, based on the reduction of the monthly water payment. Overall, RWH+ supplying higher demands with higher economics savings were feasible, as expected. However, several municipalities that showed RWH+ supplying 100% of the domestic water demands obtained lower economic savings, due to low water tariff and water consumption. Still, a set of municipalities presented RWH+ for rainwater demand replacing 50% to 60% of the residential demand, for which the high water tariffs were reflected in higher economic savings. The advantages of using the RWH systems stand out even more when the investments at Federal and Local levels are considered. HIGHLIGHTS Socioeconomic potential for rainwater harvesting in 24 Brazilian municipalities.; 10,080 system configurations were assessed for demand, rooftop area, tank capacity.; System configuration was implementable if it reached at least 80% reliability.; Economic savings of water payment were influenced by water tariff and consumption.; The advantages of using rainwater stand out when the investments are considered.;

Modelling claim frequency and claim severity are topics of great interest in property-casualty insurance for supporting underwriting, ratemaking, and reserving actuarial decisions. Standard Generalized Linear Models (GLM) frequency–severity models assume a linear relationship between a function of the response variable and the predictors, independence between the claim frequency and severity, and assign full credibility to the data. To overcome some of these restrictions, this paper investigates the predictive performance of Gradient Boosting with decision trees as base learners to model the claim frequency and the claim severity distributions of an auto insurance big dataset and compare it with that obtained using a standard GLM model. The out-of-sample performance measure results show that the predictive performance of the Gradient Boosting Model (GBM) is superior to the standard GLM model in the Poisson claim frequency model. Differently, in the claim severity model, the classical GLM outperformed the Gradient Boosting Model. The findings suggest that gradient boost models can capture the non-linear relation between the response variable and feature variables and their complex interactions and thus are a valuable tool for the insurer in feature engineering and the development of a data-driven approach to risk management and insurance.

Several lactic acid bacteria (LAB) species have been recognized as probiotics and are of considerable interest due to their potential ability to confer health benefits upon consumption. In the animal feed sector, probiotics offer an alternative to the use of antibiotic growth promoters. The preservation and incorporation of probiotics into dry products requires carefully meeting several criteria and overcoming technological challenges to maintain their functionality. Drying is a crucial step in the process, but the probiotic properties of the resulting powder and the final cell viability in the food product are significantly influenced by the type of protective compounds and drying techniques employed. In light of the growing demand for functional animal products, this review focuses on the damages incurred during microorganism dehydration processes for food incorporation, and explores strategies to minimize such damages. It provides an overview of the effects of probiotic products in the animal feed industry, including their incorporation in low-moisture food matrices and key considerations for success. Additionally, it highlights postbiotics as an attractive alternative for live probiotic cells with many technological advantages.

Abstract In this work, an experimental analysis of the performance of different types of quasi-randomly distributed reflectors inscribed into a single-mode fiber as a sensing mirror is presented. These artificially-controlled backscattering fiber reflectors are used in short linear cavity fiber lasers. In particular, laser emission and sensor application features are analyzed when employing optical tapered fibers, micro-drilled optical fibers and 50 μm-waist or 100 μm-waist micro-drilled tapered fibers (MDTF). Single-wavelength laser with an output power level of about 8.2 dBm and an optical signal-to-noise ratio of 45 dB were measured when employing a 50 μm-waist micro-drilled tapered optical fiber. The achieved temperature sensitivities were similar to those of FBGs; however, the strain sensitivity improved more than one order of magnitude in comparison with FBG sensors, attaining slope sensitivities as good as 18.1 pm/με when using a 50 μm-waist MDTF as distributed reflector.

Abstract The mojicones are traditional bread dough products that are very popular in several provinces of the central area of the Sierra. They are desired for their sweet taste and their benefits when consumed in the daily diet. This project sought to understand the bromatological composition of the mijicon to improve its commercialization in the city of Riobamba. A descriptive non-experimental design methodology was used with a correlational approach. According to the bromatological analysis, the composition in a 100g portion was as follows: protein (7.92%), fat (7.49%), total dietary fiber (5.37%), carbohydrates (73.2%), sodium chloride (6,762.22 mg), total sugars (348.7 mg), sodium (265.87%), moisture (4.45%) and ashes (1.44%). An informal survey was carried out through various social networks to examine the acceptability of the product and its pros and cons; a SWOT analysis was used. The results showed that the product has high acceptability because it is an innovative product, and its disadvantage is that capital is too weak to stock up on its advertising and marketing. It is therefore possible to recommend wider consumption of the product by a larger population, due to its substantial energy and nutritional benefits.

This article presents two applications developed using Jupyter Notebook in the Google Colab, combining several Python libraries that enable an interactive environment to query, manipulate, analyse, and visualise spatial data. The first application is from an educational context within the MAPFOR project, aiming to elaborate an interactive map of the spatial distributions of teachers with higher education degrees or pedagogical complementation per vacancies in higher education courses. The Jupyter solutions were applied in MAPFOR to better communicate within the research team, mainly in the development area. The second application is a framework to analyse and visualise collaborative emotional mapping data in urban mobility, where the emotions were collected and represented through emojis. The computational notebook was applied in this emotional mapping to enable the interaction of users, without a SQL background, with spatial data stored in a database through widgets to analyse and visualise emotional spatial data. We developed these different contexts in a Jupyter Notebook to practice the FAIR principles, promote the Open Science movement, and Open Geospatial Resources. Finally, we aim to demonstrate the potential of using a mix of open geospatial technologies for generating solutions that disseminate geographic information.

Background The increasingly widespread use of next generation sequencing protocols has brought the need for the development of user-friendly raw data processing tools. Here, we explore 2FAST2Q, a versatile and intuitive standalone program capable of extracting and counting feature occurrences in FASTQ files. Despite 2FAST2Q being previously described as part of a CRISPRi-seq analysis pipeline, in here we further elaborate on the program’s functionality, and its broader applicability and functions. Methods 2FAST2Q is built in Python, with published standalone executables in Windows MS, MacOS, and Linux. It has a familiar user interface, and uses an advanced custom sequence searching algorithm. Results Using published CRISPRi datasets in which Escherichia coli and Mycobacterium tuberculosis gene essentiality, as well as host-cell sensitivity towards SARS-CoV2 infectivity were tested, we demonstrate that 2FAST2Q efficiently recapitulates published output in read counts per provided feature. We further show that 2FAST2Q can be used in any experimental setup that requires feature extraction from raw reads, being able to quickly handle Hamming distance based mismatch alignments, nucleotide wise Phred score filtering, custom read trimming, and sequence searching within a single program. Moreover, we exemplify how different FASTQ read filtering parameters impact downstream analysis, and suggest a default usage protocol. 2FAST2Q is easier to use and faster than currently available tools, efficiently processing not only CRISPRi-seq / random-barcode sequencing datasets on any up-to-date laptop, but also handling the advanced extraction of de novo features from FASTQ files. We expect that 2FAST2Q will not only be useful for people working in microbiology but also for other fields in which amplicon sequencing data is generated. 2FAST2Q is available as an executable file for all current operating systems without installation and as a Python3 module on the PyPI repository (available at https://veeninglab.com/2fast2q).

The identification of Parkinson’s disease (PD) biomarkers has become a main goal for the diagnosis of this neurodegenerative disorder. PD has not only been intrinsically related to neurological problems, but also to a series of alterations in peripheral metabolism. The purpose of this study was to identify metabolic changes in the liver in mouse models of PD with the scope of finding new peripheral biomarkers for PD diagnosis. To achieve this goal, we used mass spectrometry technology to determine the complete metabolomic profile of liver and striatal tissue samples from WT mice, 6-hydroxydopamine-treated mice (idiopathic model) and mice affected by the G2019S-LRRK2 mutation in LRRK2/PARK8 gene (genetic model). This analysis revealed that the metabolism of carbohydrates, nucleotides and nucleosides was similarly altered in the liver from the two PD mouse models. However, long-chain fatty acids, phosphatidylcholine and other related lipid metabolites were only altered in hepatocytes from G2019S-LRRK2 mice. In summary, these results reveal specific differences, mainly in lipid metabolism, between idiopathic and genetic PD models in peripheral tissues and open up new possibilities to better understand the etiology of this neurological disorder.

Mycobacterium bovis causes tuberculosis (TB) in cattle, which in turn can transmit the pathogen to humans. Tuberculosis in dairy cattle is of particular concern where the consumption of raw milk and dairy products is customary. Baja California (BCA), Mexico, presents high prevalence of TB in both cattle and humans, making it important to investigate the molecular epidemiology of the disease in the region. A long-term study was undertaken to fully characterize the diversity of M. bovis genotypes circulating in dairy cattle, cheese and humans in BCA by whole-genome sequencing (WGS). During a 2-year period, 412 granulomatous tissue samples were collected from local abattoirs and 314 cheese samples were purchased from local stores and vendors in BCA and sent to the laboratory for mycobacterial culture, histology, direct PCR and WGS. For tissue samples M. bovis was recovered from 86.8%, direct PCR detected 90% and histology confirmed 85.9% as mycobacteriosis-compatible. For cheese, M. bovis was recovered from 2.5% and direct PCR detected 6% of the samples. There was good agreement between diagnostic tests. Subsequently, a total of 345 whole-genome SNP sequences were obtained. Phylogenetic analysis grouped these isolates into 10 major clades. SNP analysis revealed putative transmission clusters where the pairwise SNP distance between isolates from different dairies was ≤3 SNP. Also, human and/or cheese isolates were within 8.45 (range 0–17) and 5.8 SNP (range 0–15), respectively, from cattle isolates. Finally, a comparison between the genotypes obtained in this study and those reported previously suggests that the genetic diversity of M. bovis in BCA is well-characterized, and can be used to determine if BCA is the likely source of M. bovis in humans and cattle in routine epidemiologic investigations and future studies. In conclusion, WGS provided evidence of ongoing local transmission of M. bovis among the dairies in this high-TB burden region of BCA, as well as show close relationships between isolates recovered from humans, cheese, and cattle. This confirms the need for a coordinated One Health approach in addressing the elimination of TB in animals and humans. Overall, the study contributes to the knowledge of the molecular epidemiology of M. bovis in BCA, providing insight into the pathogen's dynamics in a high prevalence setting.

We recently identified acyl coenzyme A-binding protein (ACBP)/diazepam binding inhibitor (DBI) as a novel ‘hunger factor’: a protein that is upregulated in human or murine obesity and that, if administered to mice, causes hyperphagy, adipogenesis and obesity. Conversely, neutralization of ACBP/DBI by systemic injection of neutralizing monoclonal antibodies or autoantibodies produced after auto-immunization against ACBP/DBI has anorexigenic and lipolytic effects. Thus, neutralization of ACBP/DBI results in reduced food intake subsequent to the activation of anorexigenic neurons and the inactivation of orexigenic neurons in the hypothalamus. Moreover, ACBP/DBI neutralization results into enhanced triglyceride lipolysis in white fat, a surge in free fatty acids in the plasma, enhanced incorporation of glycerol-derived carbon atoms into glucose, as well as an increase in β-oxidation, resulting in a net reduction of fat mass. Importantly, ACBP/DBI neutralization also stimulated an increase in autophagy in various organs, suggesting that it might mediate anti-ageing effects.

Abstract Background Significant advances in the molecular profiling of gliomas, led the 2016 World Health Organization (WHO) Classification to include, for the first-time, molecular biomarkers in glioma diagnosis: IDH mutations and 1p/19q codeletion. Here, we evaluated the effect of this new classification in the stratification of gliomas previously diagnosed according to 2007 WHO classification. Then, we also analyzed the impact of TERT promoter mutations, PTEN deletion, EGFR amplification and MGMT promoter methylation in diagnosis, prognosis and response to therapy in glioma molecular subgroup. Methods A cohort of 444 adult gliomas was analyzed and reclassified according to the 2016 WHO. Mutational analysis of IDH1 and TERT promoter mutations was performed by Sanger sequencing. Statistical analysis was done using SPSS Statistics 21.0. Results The reclassification of this cohort using 2016 WHO criteria led to a decrease of the number of oligodendrogliomas (from 82 to 49) and an increase of astrocytomas (from 49 to 98), while glioblastomas (GBM) remained the same (n = 256). GBM was the most common diagnosis (57.7%), of which 55.2% were IDH-wildtype. 1p/19q codeleted gliomas were the subgroup associated with longer median overall survival (198 months), while GBM IDH-wildtype had the worst outcome (10 months). Interestingly, PTEN deletion had poor prognostic value in astrocytomas IDH-wildtype (p = 0.015), while in GBM IDH-wildtype was associated with better overall survival (p = 0.042) as well as MGMT promoter methylation (p = 0.009). EGFR amplification and TERT mutations had no impact in prognosis. Notably, EGFR amplification predicted a better response to radiotherapy (p = 0.011) and MGMT methylation to chemo-radiotherapy (p = 0.003). Conclusion In this study we observed that the 2016 WHO classification improved the accuracy of diagnosis and prognosis of diffuse gliomas, although the available biomarkers are not enough. Therefore, we suggest MGMT promoter methylation should be added to glioma classification. Moreover, we found two genetic/clinical correlations that must be evaluated to understand their impact in the clinical setting: i) how is PTEN deletion a favorable prognostic factor in GBM IDH wildtype and an unfavorable prognostic factor in astrocytoma IDH wildtype and ii) how EGFR amplification is an independent and strong factor of response to radiotherapy.

Cancer is the second cause of death worldwide, surpassed only by cardiovascular diseases, due to the lack of early diagnosis, and high relapse rate after conventional therapies. Chemotherapy inhibits the rapid growth of cancer cells, but it also affects normal cells with fast proliferation rate. Therefore, it is imperative to develop other safe and more effective treatment strategies, such as gene therapy, in order to significantly improve the survival rate and life expectancy of patients with cancer. The aim of gene therapy is to transfect a therapeutic gene into the host cells to express itself and cause a beneficial biological effect. However, the efficacy of the proposed strategies has been insufficient for delivering the full potential of gene therapy in the clinic. The type of delivery vehicle (viral or non viral) chosen depends on the desired specificity of the gene therapy. The first gene therapy trials were performed with therapeutic genes driven by viral promoters such as the CMV promoter, which induces non-specific toxicity in normal cells and tissues, in addition to cancer cells. The use of tumor-specific promoters over-expressed in the tumor, induces specific expression of therapeutic genes in a given tumor, increasing their localized activity. Several cancer- and/or tumor-specific promoters systems have been developed to target cancer cells. This review aims to provide up-to-date information concerning targeting gene therapy with cancer- and/or tumor-specific promoters including cancer suppressor genes, suicide genes, anti-tumor angiogenesis, gene silencing, and gene-editing technology, as well as the type of delivery vehicle employed. Gene therapy can be used to complement traditional therapies to provide more effective treatments.

We present the case of a 27-year old female with an ovarian tumor and alpha-fetoprotein (AFP) of 1210 ng/m, a right salpingo-oophorectomy was performed and had conservative complementary staging by gynecologic oncologists. The histopathological report was primary hepatoid carcinoma of the ovary (HCO), clinical stage IA, complementary treatment was adjuvant chemotherapy with BEP and remains clinical, imaging and biochemically disease free in three years follow up.

Linking pensions to longevity developments at retirement age has been one of the most common policy responses of pension schemes to aging populations. The introduction of automatic stabilizers is primarily motivated by cost containment objectives, but there are other dimensions of welfare restructuring in the politics of pension reforms, including recalibration, rationalization, and blame avoidance for unpopular policies that involve retrenchments. This paper examines the policy designs and implications of linking entry pensions to life expectancy developments through sustainability factors or life expectancy coefficients in Finland, Portugal, and Spain. To address conceptual and specification uncertainty in policymaking, we propose and apply a Bayesian model averaging approach to stochastic mortality modeling and life expectancy computation. The results show that: (i) sustainability factors will generate substantial pension entitlement reductions in the three countries analyzed; (ii) the magnitude of the pension losses depends on the factor design; (iii) to offset pension cuts and safeguard pension adequacy, individuals will have to prolong their working lives significantly; (iv) factor designs considering cohort longevity markers would have generated higher pension cuts in countries with increasing life expectancy gap.

The by-products of the food industry are an economic alternative as a source of nutrients to obtain biomass. At the same time, theiruse could solve the environmental problem related to their disposal, which is highly polluting due to their elevated biochemical oxygen demand. In this work, we seek to optimize the production of cellular biomass of two native Patagonian strains of Lactiplantibacillus plantarum (UNQLp 11 and UNQLp155), selected for its oenological and technological properties, using apple pomace (AP), a residue from the juice and cider industry. The supplementation of AP with yeast extract, salts, and Tween 80 (sAP), proved to maintain the growth of the Lpb. plantarum strains, similar to the commercial medium used to grow LAB (De Man, Rogosa, Sharpe, MRS). Cultures grown in sAP medium showed good tolerance to wine conditions (high ethanol content and low pH), demonstrated by its ability to consume L-malic acid. The subsequent inoculation of these cultures in sterile wines (Merlot and Pinot noir) was carried out at laboratory scale, evaluating cell viability and L-malic acid consumption for 21 days at 21 °C. Cultures grown in sAP media showed a similar performance to MRS media. Thus, sAP media proved to be a suitable substrate to grow oenological Lpb. plantarum strains where cultures (with high size inoculums) were able to drive malolactic fermentation, with an L-malic acid consumption higher than 90%.

In 2015, we investigated Bartonella quintana and typhus group rickettsiae in body lice from homeless persons in Bogotá, Colombia. We found B. quintana–infected body lice and seroprevalence of this microorganism in 19% of homeless persons and typhus group rickettsiae in 56%. Public health professionals should start preemptive measures and active vector control.

Objectives: To determine genetic diversity by comparing the whole genome sequences of cattle and human Mycobacterium bovis isolates from Baja California. Methods: A whole genome sequencing strategy was used to obtain the molecular fingerprints of 172 isolates of M. bovis obtained from Baja California, Mexico; 155 isolates were from cattle and 17 isolates were from humans. Spoligotypes were characterized in silico and single nucleotide polymorphism (SNP) differences between the isolates were evaluated. Results: A total of 12 M. bovis spoligotype patterns were identified in cattle and humans. Two predominant spoligotypes patterns were seen in both cattle and humans: SB0145 and SB1040. The SB0145 spoligotype represented 59% of cattle isolates (n = 91) and 65% of human isolates (n = 11), while the SB1040 spoligotype represented 30% of cattle isolates (n = 47) and 30% of human isolates (n = 5). When evaluating SNP differences, the human isolates were intimately intertwined with the cattle isolates. Conclusions: All isolates from humans had spoligotype patterns that matched those observed in the cattle isolates, and all human isolates shared common ancestors with cattle in Baja California based on SNP analysis. This suggests that most human tuberculosis caused by M. bovis in Baja California is derived from M. bovis circulating in Baja California cattle. These results reinforce the importance of bovine tuberculosis surveillance and control in this region.

Abstract Background Bovine tuberculosis (TB) control programs generally rely on the tuberculin skin test (TST) for ante-mortem detection of Mycobacterium bovis-infected cattle. Results Present findings demonstrate that a rapid antibody test based on Dual-Path Platform (DPP®) technology, when applied 1-3 weeks after TST, detected 9 of 11 and 34 of 52 TST non-reactive yet M. bovis-infected cattle from the US and GB, respectively. The specificity of the assay ranged from 98.9% (n = 92, US) to 96.0% (n = 50, GB) with samples from TB-free herds. Multi-antigen print immunoassay (MAPIA) revealed the presence of antibodies to multiple antigens of M. bovis in sera from TST non-reactors diagnosed with TB. Conclusions Thus, use of serologic assays in series with TST can identify a significant number of TST non-reactive tuberculous cattle for more efficient removal from TB-affected herds.

This paper recognizes tourism’s potential in balancing economic development of communities in the peripheries of watersheds with nature conservation. It was conducted in Ipo watershed which has 6,600 hectares of forest and provides fresh water to more than 13 million people in Metro Manila, Philippines. The objectives of the study are to determine the community’s perception of tourism and identify factors affecting their support for tourism development and environment conservation. Survey questionnaires and face to face interviews were conducted to 102 respondents. To determine the relationship of different variables, Structural Equation Modelling was used. The study revealed that the communities have positive perception on tourism and in using the Motivation, Opportunity Ability model the study revealed that motivation and ability had significant effect with support for tourism development and environment conservation. The study contributes to the literature on tourism in protected area and can be used by the national and local government in formulating strategies in achieving sustainable protected area management.

Interest in the origin and evolution of Equus dates back to over a century, but there is still no consensus on the definition of the genus or its phylogenetic position. We review the placement of Equus within several phylogenetic frameworks and present a phylogenetic analysis of derived Equini, including taxa referred to Equus, Haringtonhippus, Dinohippus, Astrohippus, Hippidion, and Boreohippidion. A new, morphology-based phylogenetic tree was used as an initial hypothesis for discussing what taxa Equus encompasses, using four criteria previously used to define the genus category in mammals: phylogenetic gaps, uniqueness of adaptive zone, crown group definition, and divergence time. According to the phylogenetic gaps criterion, Equus encompasses clade 6 (Ha. francisci = E. francisci, E. conversidens, E. quagga, E. hemionus, E. mexicanus, E. ferus, E. occidentalis, and E. neogeus) based on morphological synapomorphies. Equus is assigned to clade 6, or possibly clade 7, according to the uniqueness of adaptive zone criterion. The crown group criterion places Equus at clade 6. Based on the time-calibrated phylogeny of Equini, the divergence time criterion suggests that Equus encompasses clade 9. This clade comprises all taxa traditionally assigned to Equus analyzed in our study, including the eight taxa listed above as well as E. stenonis, E. idahoensis, and E. simplicidens; the latter two are sometimes referred to the subgenus Plesippus and the former to the subgenus Allohippus. With the exception of the divergence time criterion, the results of our evaluation are congruent in identifying clade 6 as the most suitable position for Equus. The taxonomic implications of delimiting Equus to clade 6 in our phylogenetic tree include elevation of Allohippus and Plesippus to generic rank, assignment of a new genus to “Dinohippus” mexicanus, and synonymy of Haringtonhippus with Equus.

The stomach bacterium Helicobacter pylori is one of the most prevalent pathogens in humans, closely linked with serious diseases such as gastric cancer. The microbe has been associated with its host for more than 100,000 years and escorted modern humans out of Africa. H. pylori is predominantly transmitted within families and dispersed globally, resulting in distinct phylogeographic patterns, which can be utilized to investigate migrations and bioturbation events in human history. Latin America was affected by several human migratory waves due to the Spanish colonisation that drastically changed the genetic load and composition of the bacteria and its host. Genetic evidence indicates that independent evolutionary lines of H. pylori have evolved in mestizos from Colombia and other countries in the region during more than 500 years since colonisation. The vacuolating cytotoxin VacA represents a major virulence factor of the pathogen comprising two domains, p33 and p55, the latter of which is essential for binding to the host epithelial cell. The evolution of the VacA toxin in Colombia has been strongly biased due to the effects of Spanish colonization. However, the variation patterns and microevolution of the p55 domain have not yet been described for this population. In the present study, we determined the genetic polymorphisms and deviations in the neutral model of molecular evolution in the p55 domain of 101 clinical H. pylori isolates collected in Bogotá, a city located in Andean mountains characterized by its high gastric cancer risk and its dominant mestizo population. The microevolutionary patterns of the p55 domain were shaped by recombination, purifying and episodic diversifying positive selection. Furthermore, amino acid positions 261 and 321 in the p55 domain of VacA show a high variability among mestizos clinical subsets, suggesting that natural selection in H. pylori may operate differentially in patients with different gastric diseases.

Increasing retirement ages in an automatic or scheduled way with increasing life expectancy at retirement is a popular pension policy response to continuous longevity improvements. The question addressed here is: to what extent is simply adopting this approach likely to fulfill the overall goals of policy? To shed some light on the answer, we examine the policies of four countries that have recently introduced automatic indexation of pension ages to life expectancy–The Netherlands, Denmark, Portugal and Slovakia. To this end, we forecast an alternative period and cohort life expectancy measures using a Bayesian Model Ensemble of heterogeneous stochastic mortality models comprised of parametric models, principal component methods, and smoothing approaches. The approach involves both the selection of the model confidence set and the determination of optimal weights. Model-averaged Bayesian credible prediction intervals are derived accounting for various stochastic process, model, and parameter risks. The results show that: (i) retirement ages are forecasted to increase substantially in the coming decades, particularly if a constant period in retirement is targeted; (ii) retirement age policy outcomes may substantially deviate from the policy goal(s) depending on the design adopted and its implementation; and (iii) the choice of a cohort over period life expectancy measure matters. In addition, the distributional issues arising with the increasing socio-economic gap in life expectancy remain largely unaddressed.

This research evaluates the fracture behavior of concrete with reactive magnesium oxide (MgO). Replacing cement with MgO is an attractive option for the concrete industry, mainly due to sustainability benefits and reduction of shrinkage. Four different MgO’s from Australia, Canada, and Spain were used in the concrete mixes, as a partial substitute of cement, at 5%, 10%, and 20% (by weight). The fracture toughness (KI) intensity factor and the stress–strain softening parameters of the wedge split test were evaluated after 28 days. The experimental results showed that the replacement of cement with MgO reduced the fracture energy between 13% and 53%. Moreover, the fracture energy was found to be correlated with both compressive strength and modulus of elasticity. A well-defined relationship between these properties is important for an adequate prediction of the non-linear behavior of reinforced concrete structures made with partial replacement of cement with MgO.

Protecting against unexpected yield curve, inflation, and longevity shifts are some of the most critical issues institutional and private investors must solve when managing post-retirement income benefits. This paper empirically investigates the performance of alternative immunization strategies for funding targeted multiple liabilities that are fixed in timing but random in size (inflation-linked), i.e., that change stochastically according to consumer price or wage level indexes. The immunization procedure is based on a targeted minimax strategy considering the M-Absolute as the interest rate risk measure. We investigate to what extent the inflation-hedging properties of ILBs in asset liability management strategies targeted to immunize multiple liabilities of random size are superior to that of nominal bonds. We use two alternative datasets comprising daily closing prices for U.S. Treasuries and U.S. inflation-linked bonds from 2000 to 2018. The immunization performance is tested over 3-year and 5-year investment horizons, uses real and not simulated bond data and takes into consideration the impact of transaction costs in the performance of immunization strategies and in the selection of optimal investment strategies. The results show that the multiple liability immunization strategy using inflation-linked bonds outperforms the equivalent strategy using nominal bonds and is robust even in a nearly zero interest rate scenario. These results have important implications in the design and structuring of ALM liability-driven investment strategies, particularly for retirement income providers such as pension schemes or life insurance companies.

Lactiplantibacillus plantarum strain UNQLp 11 is a lactic acid bacterium with the potential to carry out malolactic fermentation (MLF) in red wines. Recently, the complete genome of UNQLp 11 was sequenced and this strain possesses four loci of the enzyme β-glucosidase. In order to demonstrate that these glucosidase enzymes could be functional under harsh wine conditions, we evaluated the hydrolysis of p-nitrophenyl-β-D-glucopyranoside (p-NPG) in synthetic wine with different ethanol contents (0%, 12%, and 14% v/v) and at different pH values (3.2, 3.5, and 3.8). Then, the hydrolysis of precursor n-octyl β-D-glucopyranoside was analyzed in sterile Pinot Noir wine (containing 14.5% v/v of ethanol, at different pH values) by headspace sorptive extraction gas chromatography-mass spectrometry (HSSE-GC/MS). The hydrolysis of p-NPG showed that β-glucosidase activity is very susceptible to low pH but induced in the presence of high ethanol content. Furthermore, UNQLp 11 was able to release the glycosilated precursor n-octyl, during MLF to a greater extent than a commercial enzyme. In conclusion, UNQLp 11 could improve the aromatic profile of the wine by the release of volatile precursors during MLF.

Panthera atrox was a common large-sized cat in North America during the late Pleistocene. An isolated lower canine and a fifth metacarpal bone referable to this species were recovered from fluvial Quaternary deposits that outcrop in southeastern Hidalgo, central Mexico. Associated fossil material belonging to Bison indicates a Rancholabrean North American Land Mammal Age; the age assignment is corroborated by the presence of P. atrox. A comparative study with selected specimens of Panthera and Smilodon indicates that the Hidalgoan sample shares the following diagnostic features with P. atrox: a large, robust, and non-strongly curved lower canine; a large and relatively slender fifth metacarpal with a well-developed projection on the palmar side at the proximal end, narrow articulating surface for the unciform; a narrow notch on the articulating surface for the fourth metacarpal; and a diaphysis that at the middle is oval in cross section. The record supplements the evidence of P. atrox in central Mexico and represents the first reported occurrence of this cat species in the state of Hidalgo. By the same token, the known geographic distribution of P. atrox in the Mexican territory suggests that it was relatively common in temperate areas of central Mexico between 19 and 24° N at an altitudinal range from 1500 to 2250 m a.s.l. The large size (mean body mass of 300 kg) and hypercarnivorous adaptations of the American lion suggest it was the top predator of the mammalian community recorded at southeastern Hidalgo, displacing other members of the carnivore guild at the mesopredator level, such as the dire wolf (Canis dirus ), which has been also reported in the area. The high diversity of large herbivores recorded at southeastern Hidalgo, which in turn could represent potential prey of P. atrox, suggests that some areas that now are part of central Mexico were suitable hunting sites for this large-sized cat. A review of the Pleistocene record of Felidae from Mexico revealed that it encompasses 87.5 and 73.3 % of generic and specific diversity known for North America, respectively, including seven genera (Felis, Miracinonyx, Panthera, Puma, Lynx, Leopardus, and Smilodon) and 11 species (Felis rexroadensis, Miracinonyx inexpectatus, Panthera atrox, Panthera onca, Puma concolor, Puma yagouaroundi, Lynx rufus, Leopardus pardalis, Leopardus wiedii, Smilodon fatalis, and Smilodon gracilis). The majority of these taxa have been reported from numerous late Pleistocene localities; in particular, Panthera atrox was relatively common and widely distributed across the Mexican territory.

We characterized the dynamics of autophagy in vitro using four different cell systems and analyzing markers widely used in this field, i.e. LC3 (microtubule-associated protein 1 light chain 3; protein recruited from the cytosol (LC3-I) to the autophagosomal membrane where it is lipidated (LC3-II)) and p62/SQSTM1 (adaptor protein that serves as a link between LC3 and ubiquitinated substrates), (Klionsky et al., 2016) [1]. Data provided include analyses of protein levels of LC3 and p62 by Western-blotting and endogenous immunofluorescence experiments, but also p62 mRNA levels obtained by quantitative PCR (qPCR). To monitor the turnover of these autophagy markers and, thus, measure the flux of this pathway, cells were under starvation conditions and/or treated with bafilomycin A1 (Baf. A1) to block fusion of autophagosomes with lysosomes. Keywords: Autophagy, LC3, p62, Western-blot