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Resumen Introduccion El objetivo de este estudio es la caracterizacion de cepas de Streptococcus pyogenes con fenotipo mucoide y su comparacion con las cepas no mucoides aisladas entre abril y agosto de 2016. Material y metodos Se llevo a cabo la caracterizacion y el estudio de sensibilidad antimicrobiana de todos los aislados. Se determino el tipo emm y se analizaron los genes de exotoxinas speA, speB, speC, speF, speG, speH, speJ, speZ y ssa. Se recogieron datos clinicos y demograficos. Resultados De 96 aislados analizados, el 47% presentaron un fenotipo mucoide, y de estos ultimos, el 95,5% presentaron los genes speA-speB-speF-speG-ssa y genotipo emm3. La principal manifestacion clinica entre todos los pacientes fue faringoamigdalitis (77,1%) que evoluciono a escarlatina en el 67,5% de los casos. Conclusion Se describe la circulacion de una cepa de aspecto mucoide con perfil de toxinas speA-speB-speF-speG-ssa y genotipo emm3.1 considerado de los mas frecuentes y mas virulentos de SGA.

Egyptian Baladi cattle is one of the few native taurine breeds which remain in the original domestication area of Bos taurus, the Nile Valley (Upper Egypt). Apart from its evolutive relevance, the ...

The goal of this research was to evaluate the nutritive value of full-fat de-hulled sunflower seeds (DHSS) in feeding broiler chickens. Three experiments were conducted. Experiment 1 determined the proximal analysis of DHSS, the essential amino acid content, and apparent metabolizable energy (AME). In experiment 2, two diets were formulated, namely a control and a diet containing 10% DHSS. Apparent ileal digestibility (AID) of dry matter (DM), crude protein (CP), and ether extract (EE) were calculated. In experiment 3, the performances of broilers fed the two diets were evaluated in terms of the relative weights of digestive system organs, breast meat colour, growth and feed consumption. Full-fat de-hulled sunflower seeds were characterized as having 507.9 g/kg EE, 282.1 g/kg CP, 21.02 MJ/kg AME and 9.3 g/16 g N arginine. Apparent ileal digestibility of DM in the 10% DHSS diet was 658.0 g/kg, similar to that of the control diet (643.0 g/kg). Partial substitution of DHSS for soybean meal (SBM) did not affect the relative weight of digestive system organs, except for the liver. The relative weights of the liver were 17.90 g/kg bodyweight in birds fed the DHSS diet and 16.37 g/kg bodyweight (P

Un total de 46 ovejas nulíparas Katahdin se asignaron bajo un diseño de bloques completamente al azar a dos tratamientos dietarios (n= 23) alrededor del empadre para evaluar el efecto de adicionar ácidos grasos poliinsaturados (AGP) sobre el comportamiento reproductivo y el crecimiento pre-destete de las crías. Los tratamientos consistieron en dos concentrados isoenergéticos e isoproteícos formulados sin (testigo) y con AGP omega 6, los cuales se ofrecieron durante 42 d (7 d antes y 35 d de exposición al semental). La adición dietaria de AGP disminuyó (P0.05) las demás variables reproductivas. En crías, el peso al nacimiento no varió (P>0.05) con la adición de AGP; sin embargo, la inclusión dietaria de AGP aumentó (P

Begonia is an important horticultural plant group, as well as one of the most speciose Angiosperm genera, with over 2000 described species. Genus wide studies of genome size have shown that Begonia has a highly variable genome size, and analysis of paralog pairs has previously suggested that Begonia underwent a whole genome duplication. We address the contribution of gene duplication to the generation of diversity in Begonia using a multi-tissue RNA-seq approach. We chose to focus on chalcone synthase (CHS), a gene family having been shown to be involved in biotic and abiotic stress responses in other plant species, in particular its importance in maximising the use of variable light levels in tropical plants. We used RNA-seq to sample six tissues across two closely related but ecologically and morphologically divergent species, Begonia conchifolia and B. plebeja, yielding 17,012 and 19,969 annotated unigenes respectively. We identified the chalcone synthase gene family members in our Begonia study species, as well as in Hillebrandia sandwicensis, the monotypic sister genus to Begonia, Cucumis sativus, Arabidopsis thaliana, and Zea mays. Phylogenetic analysis suggested the CHS gene family has high duplicate turnover, all members of CHS identified in Begonia arising recently, after the divergence of Begonia and Cucumis. Expression profiles were similar within orthologous pairs, but we saw high inter-ortholog expression variation. Sequence analysis showed relaxed selective constraints on some ortholog pairs, with substitutions at conserved sites. Evidence of pseudogenisation and species specific duplication indicate that lineage specific differences are already beginning to accumulate since the divergence of our study species. We conclude that there is evidence for a role of gene duplication in generating diversity through sequence and expression divergence in Begonia.

Electronic Supplementary Information (ESI) available: Supplementary figures and tables and copies of the 1H and 13C NMR (1D and 2D) spectra of the new compounds., The first fluorescent probes that are actively channeled into the mitochondrial matrix by a specific mitochondrial membrane transporter in living cells have been developed. The new functional probes (BCT) have a minimalist structural design based on the highly efficient and photostable BODIPY chromophore and carnitine as a biotargeting element. Both units are orthogonally bonded through the common boron atom, thus avoiding the use of complex polyatomic connectors. In contrast to known mitochondria-specific dyes, BCTs selectively label these organelles regardless of their transmembrane potential and in an enantioselective way. The obtained experimental evidence supports carnitine-acylcarnitine translocase (CACT) as the key transporter protein for BCTs, which behave therefore as acylcarnitine biomimetics. This simple structural design can be readily extended to other structurally diverse starting FBODIPYs to obtain BCTs with varied emission wavelengths along the visible and NIR spectral regions and with multifunctional capabilities. BCTs are the first fluorescent derivatives of carnitine to be used in cell microscopy and stand as promising research tools to explore the role of the carnitine shuttle system in cancer and metabolic diseases. Extension of this approach to other small-molecule mitochondrial transporters is envisaged., We gratefully acknowledge the Spanish Ministerio de Ciencia, Innovación y Universidades (MCIU), Agencia Estatal de Investigación (AEI), and European Regional Development Fund (ERDF) (projects MAT2017-83856-C3-1-P, MAT2017-83856-C3-3-P, and MAT2015-68837-REDT), Gobierno Vasco (IT912-16), and the Instituto de Salud Carlos III-Fondo de Investigación Sanitaria (CIBERONC and PI17/01901) for financial support. A. B.-M. and R. P. M. thank MCIU, AEI and ERDF, and UPV-EHU for a FPI predoctoral contract and predoctoral fellowship, respectively

Background Chagas disease is considered important and presents intense inflammatory and fibrotic processes induced by the perpetuation of the parasite in the affected tissues and organs. Therefore, it is necessary to inquire about the host defense and attack mechanisms to have a more detailed knowledge about Chagas disease. MicroRNAs are found in blood, tissues and extracellular vesicles. These small regulators of gene expression are involved in physiological and pathological processes in both mammals and parasites. Several microRNAs have deregulated expression in chagasic heart disease, although little is known about their extracellular expression. Our main objective was to evaluate the involvement of miR-21, miR-146a and miR-155 in several samples from mice infected with the TcI Ninoa strain from the acute and indeterminate phases. We also explored a potential functional association of the selected microRNAs using STRING software. This software identified 23 pathways associated with Trypanosoma cruzi infection. In addition, eleven genes were identified through bioinformatics analysis, and we found that SMAD family member 5 was downregulated in both phases. This gene serves as a mediator in the TGF-β signaling pathway. Thus, forty female mice of the CD1 strain were distributed into 4 groups and the expression levels of miR-21, miR-146a and miR-155 were measured in samples of heart tissue, total plasma and plasma extracellular vesicles by quantitative real-time polymerase chain reaction. Results Overexpression of miR-21, miR-146a and miR-155 was observed in heart and plasma in both phases. Moreover, in extracellular vesicles miR-21 and miR-146a were also overexpressed in the acute phase, whereas in the indeterminate chronic phase we found only miR-146a up-regulated. Conclusions The expression of inflammatory microRNAs miR-21, miR-146a and miR-155 were up-regulated in each of the samples from acutely and chronically infected mice. The relevant finding was that miR-146a was up-regulated in each sample in both phases; therefore, this miRNA could be a possible candidate biomarker in Chagas disease.

TCF7L2 is the most potent locus for type 2 diabetes (T2D) risk and the first locus to have been robustly reported by genomic linkage studies. TCF7L2 is a transcription factor that forms a basic part of the Wnt signaling pathway. This gene has highly conserved sequence regions that correspond to functional domains. The association of TCF7L2 with T2D is one of the most powerful genetically discovered in studies of complex diseases, as it has been consistently replicated in multiple populations with diverse genetic origins. The mechanisms over which TCF7L2 exerts its effect on T2D are still not well understood. In this article, we describe the main molecular mechanisms of how TCF7L2 is related to T2D. TCF7L2 variants associated with T2D risk exert an influence on the initial therapeutic success of the hypoglycemic oral agent sulfonylurea. Thus, it is important to know whether there are other TCF7L2 variants associated with T2D that can influence treatment with oral hypoglycemic agents. Resequencing of the TCF7L2 gene in diverse ethnic groups is required to reveal common and rare variations and their role in different pathologies and in adverse reactions to drugs. Identification of TCF7L2-susceptibility disease variants will permit, at a given moment, offering of therapies to patients according to their genotype.

Previous studies have described the clinical, serological and pathological features of patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and antibodies directed against the paranodal proteins neurofascin-155, contactin-1 (CNTN1), contactin-associated protein-1 (Caspr1), or nodal forms of neurofascin. Such antibodies are useful for diagnosis and potentially treatment selection. However, antibodies targeting Caspr1 only or the Caspr1/CNTN1 complex have been reported in few patients with CIDP. Moreover, it is unclear if these patients belong to the same pathophysiological subgroup. Using cell-based assays in routine clinical testing, we identified sera from patients with CIDP showing strong membrane reactivity when both CNTN1 and Caspr1 were co-transfected (but not when CNTN1 was transfected alone). Fifteen patients (10 male; aged between 40 and 75) with antibodies targeting Caspr1/CNTN1 co-transfected cells were enrolled for characterization. The prevalence of anti-Caspr1/CNTN1 antibodies was 1.9% (1/52) in the Sant Pau CIDP cohort, and 4.3% (1/23) in a German cohort of acute-onset CIDP. All patients fulfilled European Federation of Neurological Societies/Peripheral Nerve Society (EFNS/PNS) definite diagnostic criteria for CIDP. Seven (47%) were initially diagnosed with Guillain-Barré syndrome due to an acute-subacute onset. Six (40%) patients had cranial nerve involvement, eight (53%) reported neuropathic pain and 12 (80%) ataxia. Axonal involvement and acute denervation were frequent in electrophysiological studies. Complete response to intravenous immunoglobulin was not observed, while most (90%) responded well to rituximab. Enzyme-linked immunosorbent assay (ELISA) and teased nerve fibre immunohistochemistry confirmed reactivity against the paranodal Caspr1/CNTN1 complex. Weaker reactivity against Caspr1 transfected alone was also detected in 10/15 (67%). Sera from 13 of these patients were available for testing by ELISA. All 13 samples reacted against Caspr1 by ELISA and this reactivity was enhanced when CNTN1 was added to the Caspr1 ELISA. IgG subclasses were also investigated by ELISA. IgG4 was the predominant subclass in 10 patients, while IgG3 was predominant in other three patients. In conclusion, patients with antibodies to the Caspr1/CNTN1 complex display similar serological and clinical features and constitute a single subgroup within the CIDP syndrome. These antibodies likely target Caspr1 primarily and are detected with Caspr1-only ELISA, but reactivity is optimal when CNTN1 is added to Caspr1 in cell-based assays and ELISA.

Antibiotics are added to semen extenders to control the growth of bacteria contaminating semen during collection but may contribute to the development of antibiotic resistance. An alternative would be physical separation of spermatozoa from bacteria. The objective of the present study was to evaluate two low densities of Porcicoll for removal of bacteria, and for their effect on sperm recovery and sperm quality. Semen was collected from boars at a commercial station. Aliquots of 8 extended ejaculates were subjected to colloid centrifugation through 20% Porcicoll (P20) and 30% Porcicoll (P30) in 500 mL tubes and then stored at 17 °C. Microbiological examination and sperm quality evaluation (computer assisted sperm analysis and flow cytometry) were carried out on controls and all colloid-selected samples immediately after preparation and again after storage for 3 and 7 days. The microorganisms found were mainly bacteria from the environment, gut or skin. There was a considerable reduction or complete removal of some bacteria by both colloids. Recovery rates were 86% for P20 and 81% for P30. Sperm quality was not adversely affected by colloid centrifugation on day 0, and thereafter showed a more gradual deterioration in colloid centrifuged samples than in controls, possibly due to lower bacterial contamination. There were no differences in sperm quality between the two colloid treatments. Thus, these results show that contaminating bacteria in semen can be controlled by centrifugation through low density colloids.

Individualized pre-operative assessment of the patterns of the lower extremity anatomy and deformities in patients undergoing total knee arthroplasty seems essential for a successful surgery. In the present study, we investigated the relationship among the coronal alignment and the rotational profile of the lower extremities in the Caucasian population with end-stage knee osteoarthritis. We conducted a prospective study of 385 knees that underwent a pre-operative three-dimensional computed tomography-based model. The lower extremity alignment was determined (mechanical tibiofemoral or hip-knee-ankle angle, supplementary angle of the femoral lateral distal angle, and proximal medial tibial angle). For each case, the femoral distal rotation (condylar twist angle), the femoral proximal version, and the tibial torsion were determined. As the coronal alignment changed from varus to valgus, the femoral external rotation increased (r = 0.217; p

Metagenomics and single‐cell genomics have enabled the discovery of relevant uncultured microbes. Recently, single‐virus genomics (SVG), although still in an incipient stage, has opened new avenues in viral ecology by allowing the sequencing of one single virus at a time. The investigation of methodological alternatives and optimization of existing procedures for SVG is paramount to deliver high‐quality genomic data. We report a sequencing dataset of viral single‐amplified genomes (vSAGs) from cultured and uncultured viruses obtained by applying different conditions in each SVG step, from viral preservation and novel whole‐genome amplification (WGA) to sequencing platforms and genome assembly. Sequencing data showed that cryopreservation and mild fixation were compatible with WGA, although fresh samples delivered better genome quality data. The novel TruPrime WGA, based on primase‐polymerase features, and WGA‐X employing a thermostable phi29 polymerase, were proven to be with sufficient sensitivity in SVG. The Oxford Nanopore (ON) sequencing platform did not provide a significant improvement of vSAG assembly compared to Illumina alone. Finally, the SPAdes assembler performed the best. Overall, our results represent a valuable genomic dataset that will help to standardized and advance new tools in viral ecology. This work has been supported by Gordon and Betty Moore Foundation (grant 5334) and Spanish Ministry of Economy and Competitiveness (refs CGL2013‐40564‐R, RTI2018‐094248‐B‐I00 and SAF2013‐49267‐EXP). Work at CRG, BIST and UPF was in part funded by the Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Severo Ochoa 2013‐2017’ and the Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Maria de Maeztu 2016‐2019’.

Introduction: Avocado exports have been affected by the presence of Hemiberlesia lataniae, the main damage is aesthetic and undermines fruit quality. Objective: To determine the spatial distribution of Hemiberlesia lataniae in a ‘Hass’ avocado crop in Estado de Mexico. Methodology: The experiment was developed in three avocado growing areas (Coatepec Harinas, Donato Guerra and Ixtapan del Oro). Sampling was carried out using the quadrat methodology, a total of 400 trees were sampled per zone from August 2019 to July 2020. The trees were divided into three strata (high, medium and low), and a terminal branch from each cardinal point was marked. Each tree was marked and georeferenced with a differential GPS. Results: A total of 24 samples per zone were generated, and all cases had an aggregated spatial behavior of Hemiberlesia lataniae. Semivariograms were fitted to spherical, exponential, and Gaussian models. Limitation of the study: Geostatistics is the only method that provides the exact location of the pest and the sample in density maps. Originality: The present study proposes to implement integrated management strategies to reduce ecological damage, due to indiscriminate use of insecticides, and presence of residues in fruits. Conclusions: The following study allowed us to know the spatial distribution of Hemiberlesia lataniae by means of density maps in three ‘Hass’ avocado producing areas in Estado de Mexico, where an aggregated spatial distribution was observed.

Introduction Acinetobacter is a genus that comprises a group of opportunistic pathogens responsible for a variety of nosocomial infections. The Acinetobacter calcoaceticus–Acinetobacter baumannii (Acb) complex includes some species of clinical importance, mainly A. baumannii, A. pittii and A. nosocomialis, which share phenotypic similarities that make it very difficult to distinguish between them using a phenotypic approach. The aim of this study was to evaluate two commercial matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems for the identification of different Acinetobacter species, with a special focus among those belonging to the Acb complex. Methods One hundred and fifty-six Acinetobacter spp. clinical strains, identified by amplified ribosomal DNA restriction analysis (ARDRA) and rpoB gene sequencing, were analysed by two different MALDI-TOF systems. Results Considering only the 144 strains of the Acb complex evaluated in this study, the Vitek-MS™ and Microflex LT™ systems correctly identified 129 (89.6%) and 143 (99.3%) strains, respectively. Conclusion After analysing 156 strains belonging to Acinetobacter spp., both Vitek-MS™ and Microflex LT™ proved to be rapid and accurate systems for the identification of Acb complex species showing a good correlation. However, both manufacturers should improve their databases to include new species in them.

Domestic turkey production generally utilises only a few genetically improved lines, and local breeds are severely endangered as a result. Furthermore, the genetic resources of domestic turkeys have not been properly investigated, which could, ultimately, lead to the extinction of local breeds and negatively affect their corresponding genetic diversity and environmental adaptation. Although, several microsatellite markers have been designed for mapping and quantitative trait locus analysis, there is no standard panel of markers for genetic characterisation or genetic diversity assessment. Accordingly, the present study aimed to develop a set of polymorphic microsatellite markers that could be used for international turkey population studies. Thirty-nine microsatellites were selected based on polymorphism, DNA sequence and chromosome position, as well as on amplification efficiency, success rate and the absence of nonspecific amplification. The markers were screened using 105 DNA samples from local turkey breeds from Mexico, the United States, Italy, Brazil, Egypt and Spain. A total of 401 alleles were identified, with a mean number of alleles per marker of 10.28 ± 4.25. All microsatellites were polymorphic, with at least four alleles and no more than 19 alleles. Furthermore, allelic richness ranged from 3.810 to 17.985, mean heterozygosity ranged from 0.452 ± 0.229 to 0.667 ± 0.265, polymorphic information content values ranged from 0.213 (MNT264) to 0.850 (RHT0024) and the mean Fis value was 0.322. Overall, the panel was highly polymorphic and exhibited moderate Hardy–Weinberg disequilibrium, thereby indicating its value as a tool for biodiversity and population structure studies that could play an important role in promoting the conservation of local turkey breeds.Highlights Important genetic resources reside within indigenous turkey populations. These are linked to historic heritage production values and breeds. It is important to preserve this heritage and genetic diversity, which threatens to be lost as production systems focus on production characteristics. Microsatellite markers, even though, they are now replaced by single nucleotide polymorphism automatic genotyping platforms in many fields of genetics, remain a viable alternative thanks to their cheapness and simplicity of study which makes them particularly useful when the population to be studied lacks information of the prior genetic structure.

Objectives Corynebacterium urealyticum is a non-diphtherial urease-producing clinically relevant corynebacterium associated with urinary tract infections. Most clinical C. urealyticum isolates are multidrug-resistant. Whole-genome sequencing (WGS) of C. urealyticum VH4248 isolated from a clinical urine sample at Hospital Universitario Marques de Valdecilla, Santander, Spain, was performed to predict its antimicrobial resistance profile and to compare it with results of culture-based phenotypic antimicrobial susceptibility testing. Methods Classical microbiological methods and VITEK® MS were used for isolation and initial identification of strain VH4248. Draft genome sequencing was performed on an Illumina HiSeq 2500 platform, followed by assembly and annotation using SPAdes and RAST. Resistance genes were identified through PATRIC, the Pathosystems Resource Integration Center. Average nucleotide identity (ANI) analysis was done using the EDGAR and OrthoANI databases. Antimicrobial susceptibility was determined by Etest. Results Isolate VH4248 was initially identified asC. urealyticum. Its genome size is 2 261 231 bp with 64.4% GC content. Genome-based identification tools showed an average 93.7% similarity between VH4248 and C. urealyticum genomes deposited in public databases. Therefore, this isolate must be classified as Corynebacterium sp. The blaA and ermX genes as well as a class 1 integron including the aadB and sul1 genes are present in the VH4248 genome. This isolate is highly resistant to ampicillin, erythromycin and trimethoprim/sulfamethoxazole, and moderately resistant to gentamicin and kanamycin. Conclusions WGS is a powerful tool forCorynebacterium identification to species level and for detection of unusual resistance determinants, such as that encoded by the class 1 integron in isolate VH4248.

Here, the authors present the clinical case of a 45-year-old woman with a diagnosis of a chronic periodontal abscess of the tooth #8. After atraumatic dental extraction and bone regeneration with the use of platelet-rich fibrin and bone graft, the area showed excellent bone regeneration with adequate stability of the soft tissue, even 4 months after the surgery.

Plants respond to adverse environmental cues by adjusting a wide variety of processes through highly regulated mechanisms to maintain plant homeostasis for survival. As a result of the sessile nature of plants, their response, adjustment and adaptation to the changing environment is intimately coordinated with their developmental programs through the crosstalk of regulatory networks. Germination is a critical process in the plant life cycle, and thus plants have evolved various strategies to control the timing of germination according to their local environment. The mechanisms involved in these adjustment responses are largely unknown, however. Here, we report that mutations in core elements of canonical RNA-directed DNA methylation (RdDM) affect the germination and post-germination growth of Arabidopsis seeds grown under salinity stress. Transcriptomic and whole-genome bisulfite sequencing (WGBS) analyses support the involvement of this pathway in the control of germination timing and post-germination growth under salinity stress by preventing the transcriptional activation of genes implicated in these processes. Subsequent transcriptional effects on genes that function in relation to these developmental events support this conclusion.

Objective The objective of this study was to determine and compare the distribution of fimA genotypes of Porphyromonas gingivalis (P. gingivalis) in systemic lupus erythematosus (SLE) patients compared with control subjects. Material and methods This observational cross-sectional study included 281 patients divided into two groups. Group 1 (G1) consisted of 162 control subjects (30–54 years old) and, group 2 (G2) included 119 subjects (10–69 years old) diagnosed with SLE. The presence of P. gingivalis was detected by PCR. DNA sequences in acquired plaque samples were identified using P. gingivalis specific sequences and further analyzed to differentiate their fimA genotypes using six sets of fimA genotype-specific primers. Results The presence of periodontitis (PE) was similar in both groups; similar measurements were obtained regarding clinical attachment loss (CAL) (G1 1.76 ± 0.72 vs. G2 1.95 ± 0.76). G2 showed the highest frequency of P. gingivalis (94.95%). FimA genotype II is considered the most virulent and, was the most frequently found in the SLE group (53.09%). Conclusion The genotypes associated with PE are more frequently detected in SLE, which could make them susceptible to develop PE.

OBJECTIVES Multiple sclerosis (MS) is a neurodegenerative chronic inflammatory. Mutations in the vitamin D receptor (VDR) gene can substantially affect serum vitamin D levels or alter its functionality, and can consequently increase susceptibility to developing MS. The objective of this study was to evaluate the association between polymorphisms in the VDR gene and risk of MS in a (Spanish) Caucasian population. PATIENTS AND METHODS We conducted a retrospective case-control study comprising 209 patients with relapsing-remitting multiple sclerosis (RRMS) and 836 controls of Caucasian origin from southern Spain. The ApaI (rs7975232), BsmI (rs1544410), Cdx2 (rs11568820), FokI (rs2228570), and TaqI (rs731236) gene polymorphisms were determined by allelic discrimination real-time PCR using TaqMan probes. RESULTS The recessive logical regression model, adjusted for age and sex, revealed that the TT genotype for VDR FokI (rs2228570) polymorphism was associated with higher risk of MS (P = 0.0150; OR = 1.82; 95% CI = 1.12-2.94; TT vs. CT + CC). No association between the other polymorphisms and development of MS was found in any of the models analyzed. The haplotype analysis, adjusted for age, smoking, and sex, did not find any statistically significant association between the haplotypes analyzed and risk of MS. CONCLUSIONS The VDR FokI (rs2228570) polymorphism was significantly associated with developing MS. We found no influence of the ApaI (rs7975232), BsmI (rs1544410), Cdx2 (rs11568820), FokI (rs2228570), and TaqI (rs731236) gene polymorphisms on the risk of developing MS in our patients.

El garbanzo es una leguminosa, que se cultiva en dos regiones de México principalmente, noroeste (Sonora, Sinaloa y Baja california) y la región de El Bajío (Guanajuato, Michoacán y Jalisco); sin embargo, cada año la producción del cultivo está comprometida con la fusariosis vascular, una de las principales enfermedades que afectan al cultivo y que está asociada al complejo fúngico Fusarium oxysporum, Fusarium solani, Rhizoctonia solani, Macrophomina phaseolina y Sclerotium rolfsii. Una alternativa de control biológico es la aplicación de Trichoderma, la que además tiene un efecto indirecto en la nutrición de la planta. El objetivo de este estudio fue determinar el antagonismo in vitro de dos cepas de Trichoderma harzianum (T1 y T2) y su efecto como biofertilizante. Se realizaron confrontaciones in vitro contra cepas de las razas de Fusariumoxysporum f. sp. ciceris (Foc 0, 1B/C, 5 y 6), Fusarium solani, Macrophomina phaseolina (MSonora y M-GTO) y Sclerotium rolfsii. Se evaluó el efecto de T2 como biofertilizante (TB) midiendo las variables: número de flores, vainas, altura de la planta, diámetro del tallo, longitud de la raíz y rendimiento de grano. Las dos cepas de T. harzianum mostraron antagonismo en diferente escala contra los patógenos. Adicionalmente, con el tratamiento donde se aplicó T. harzianum (TB) se presentaron incrementos en el número de flores (30%), vainas (24%), altura (3%), diámetro de las plantas (3.5%), así como la longitud de la raíz (13%) y rendimiento del grano (23%).

The effect of partial replacement of wheat flour with turmeric flour (Curcuma longa) on the physicochemical and sensory properties of chorizo made with pork was evaluated. Three levels of turmeric flour substitution were proposed 0.8%, 1.4% and 2.1%, corresponding to a substitution of a 19.04%, 33.33% and 50,00%, respectively, along with a control chorizo without the addition of said flour. The pH, water holding capacity, color, texture and sensory evaluation were measured. The pH tended towards acidity in the treatment with the greatest inclusion of uremic flour. The sensory evaluation was carried out with to untrained panelists who determined the chorizo with the highest acceptability. For the sensory parameters, there were no significant differences, except for color. Therefore, turmeric flour can be used as a partial substitute for wheat flour in the production of common chorizo without affecting physicochemical and sensory properties. Resumen Se evaluó el Efecto del remplazo parcial de la harina de trigo por harina de cúrcuma (Curcuma longa) sobre las propiedades fisicoquímicas y sensoriales de chorizos hechos con carne de cerdo. Se propusieron tres niveles de sustitución de harina de cúrcuma 0,8%, 1,4% y 2,1%, correspondientes a una sustitución de 19,04%, 31,33% y 50,00%. Respectivamente y un chorizo control sin adición de dicha harina. Se midieron el pH, la capacidad de retención de humedad, el color, la textura y la evaluación sensorial. El pH mostró una tendencia a la acidez en el tratamiento con una mayor inclusión de harina de cúrcuma. La evaluación sensorial se trabajó con 70 panelistas no entrenados que determinaron el chorizo con la mayor aceptabilidad. En cuanto, a los parámetros sensoriales, no hubo diferencias significativas, excepto para el color. Por lo tanto, se puede concluir que la harina de cúrcuma se puede usar como un sustituto parcial de la harina de trigo en la producción de chorizo común, sin afectar sus propiedades fisicoquímicas y sensoriales.

La gran mayoría de las calabazas que se consumen en el mundo tienen su origen en especies que fueron domesticadas en México, todas ellas pertenecen el género Cucurbita. Se obtuvieron colectas de semillas provenientes de la huasteca, las localidades son: Cd. Mante, Antiguo Morelos, Cd. Valles, Río Verde y Huejutla. En relación con los antecedentes, para esta investigación se decidió escudriñar por la hibridación en cruza simple y autofecundación con selección individual de frutos. Siendo calabazas de clima cálido iniciamos y realizamos las cruzas en un invernadero con temperaturas controladas de 25 a 35 grados, en el CEVAMEX, en 2015 se iniciaron un cruzamiento directo de todas contra todas en las que tuviéramos flores para polinizar y se observó que los estambres se formaban filamentos un pie y discos nectarífero, también con diferente número 3, 4, 5 y hasta 6. El mejoramiento genético con autofecundaciones y selección individual de frutos, por lo cual tratamos de emparejar los estigmas y los estambres iguales, actualmente estamos en F3 que es una endogamia de 87.5%. Se observa en la Figura 4 se ilustra el avance del aumento de carrilleras de semillas de calabaza y eliminando los espacios dentro de las cavidades de fruto de calabaza, así mismo se observa en el Cuadro 1, los pesos de los frutos y sus rendimientos en semilla seca y el porcentaje de ganancia de semilla dentro del fruto, el de 4 carrilleras de 5 a 53% y las de 5 carrilleras es de 37 a 53%.

Healthy Wistar rats were supplemented during 20 weeks with commercial inulin (I) and Agave tequilana fructans (CAT), experimental fructans from A. tequilana (EAT) and A. salmiana (AS) mature stems, rice starch 10% (RS), and standard feed for rodents (C). Feed intake was kept steady, but with I, body weight and abdominal adipose tissue (6.01 g) decreased at the end. Glucose (mg/dL) (C, 120.52; I, 110.69; CAT, 105.75; EAT, 115.48; AS, 101.63; and RS, 121.82), total cholesterol (C, 89.89; I, 64.48; CAT, 68.04; EAT, 68.74; AS, 68.04; and RS, 82), and triglycerides (C, 84.03; I, 59.52; CAT, 68.56; EAT, 59.08; AS, 75.27; and RS, 81.8) kept being normal and without differences between fructans. At the end, there was a significant increase in lactic acid bacteria when the I and AS groups were compared to the C group (C, 9.18; I, 10.64; CAT, 10.34; EAT, 10.36; AS, 10.49; and RS, 9.62 log 10 CFU/g of feces). In addition, with fructans, there was an accelerated process in feces emptiness, Lieberkühn crypts kept their morphology, and there was an increment of goblet cells.

Background: The LRP1 (CR9) domain and, in particular, the sequence Gly1127-Cys1140 (P3) plays a critical role in the binding and internalization of aggregated LDL (agLDL). We aimed to evaluate whether immunization with P3 reduces high-fat diet (HFD)-induced atherosclerosis. Methods: Female New Zealand White (NZW) rabbits were immunized with a primary injection and four reminder doses (R1-R4) of IrP (irrelevant peptide) or P3 conjugated to the carrier. IrP and P3-immunized rabbits were randomly divided into a normal diet group and a HFD-fed group. Anti-P3 antibody levels were determined by ELISA. Lipoprotein profile, circulating and tissue lipids, and vascular pro-inflammatory mediators were determined using standardized methods while atherosclerosis was determined by confocal microscopy studies and non-invasive imaging (PET/CT and Doppler ultrasonography). Studies treating human macrophages (hMΦ) and coronary vascular smooth muscle cells (hcVSMC) with rabbit serums were performed to ascertain the potential impact of anti-P3 Abs on the functionality of these crucial cells. Results: P3 immunization specifically induced the production of anti-P3 antibodies (Abs) and did not alter the lipoprotein profile. HFD strongly induced cholesteryl ester (CE) accumulation in the aorta of both the control and IrP groups, and their serum dose-dependently raised the intracellular CE of hMΦ and hcVSMC, promoting TNFR1 and phospho-NF-kB (p65) overexpression. These HFD pro-inflammatory effects were dramatically decreased in the aorta of P3-immunized rabbits and in hMΦ and hcVSMC exposed to the P3 rabbit serums. Microscopy studies revealed that P3 immunization reduced the percentage of lipids, macrophages, and SMCs in the arterial intima, as well as the atherosclerotic extent and lesion area in the aorta. PET/CT and Doppler ultrasonography studies showed that the average standardized uptake value (SUVmean) of the aorta and the arterial resistance index (ARI) of the carotids were more upregulated by HFD in the control and IrP groups than the P3 group. Conclusions: P3 immunization counteracts HFD-induced fatty streak formation in rabbits. The specific blockade of the LRP1 (CR9) domain with Anti-P3 Abs dramatically reduces HFD-induced intracellular CE loading and harmful coupling to pro-inflammatory signaling in the vasculature., The main support to develop this project was received from Fundació MARATÓ TV3 Project 201521-10 (to VLl-C) and FIS PI18/01584 (to VLl-C), FIS16/00139 (to JCE-G) from the Instituto de Salud Carlos III (ISCIII) and co-financed with ERDFs. Support was received from Fundació Marató TV3 (Project 201521-10) for post-doctoral funding to OB and from Ministerio de Economía y Competitividad to DdG-C (IJCI-2016-29393). CIBER Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM; CB07/08/0016 to JCE-G) and CIBER Enfermedades Cardiovasculares (CIBERCV; CB16/1100403 to DdGC, VLl-C and CB16/11/00411 to LB and MB-P) are projects run by the Instituto de Salud Carlos III (ISCIII). We also acknowledge support from “Secretaria d’Universitats i Recerca del Departament d’Economia I Coneixement de la Generalitat de Catalunya” [2017SGR946 to VLl-C and 2017SGR1480 to LB]. DdG-C, VL-C and JCE-G are members of the Group of Vascular Biology of the Spanish Society of Atherosclerosis (SEA).

Background and ObjectivesTo study the clinical and laboratory features of antineurofascin-155 (NF155)–positive autoimmune nodopathy (AN).MethodsPatients with anti-NF155 antibodies detected on routine immunologic testing were included. Clinical characteristics, treatment response, and functional scales (modified Rankin Scale [mRS] and Inflammatory Rasch-built Overall Disability Scale [I-RODS]) were retrospectively collected at baseline and at the follow-up. Autoantibody and neurofilament light (NfL) chain levels were analyzed at baseline and at the follow-up.ResultsForty NF155+ patients with AN were included. Mean age at onset was 42.4 years. Patients presented with a progressive (75%), sensory motor (87.5%), and symmetric distal-predominant weakness in upper (97.2%) and lower extremities (94.5%), with tremor and ataxia (75%). Patients received a median of 3 (2–4) different treatments in 46 months of median follow-up. Response to IV immunoglobulin (86.8%) or steroids (72.2%) was poor in most patients, whereas 77.3% responded to rituximab. HLA-DRB1*15 was detected in 91.3% of patients. IgG4 anti-NF155 antibodies were predominant in all patients; anti-NF155 titers correlated with mRS within the same patient (r = 0.41, p = 0.004). Serum NfL (sNfL) levels were higher in anti-NF155+ AN than in healthy controls (36.47 vs 7.56 pg/mL, p < 0.001) and correlated with anti-NF155 titers (r = 0.43, p = 0.001), with I-RODS at baseline (r = −0.88, p < 0.001) and with maximum I-RODS achieved (r = −0.58, p = 0.01). Anti-NF155 titers and sNfL levels decreased in all rituximab-treated patients.DiscussionAnti-NF155 AN presents a distinct clinical profile and good response to rituximab. Autoantibody titers and sNfL are useful to monitor disease status in these patients. The use of untagged-NF155 plasmids minimizes the detection of false anti-NF155+ cases.Classification of EvidenceThis study provides Class IV evidence that anti-NF155 antibodies associate with a specific phenotype and response to rituximab.

The objective was to determine the effect of the cutting moment on the productive response of three genotypes of L. corniculatus. The design was completely randomized, with three replications. The highest and lowest yields were presented at 35 and 7 DDR with 5.48 and 0.72 g DM plant-1. The blade was the one that contributed the most to the performance. The highest and lowest leaf:stem ratio was recorded at 7 (5.71) and 42 RDD (2.39), with the 255301 genotype being superior to the rest p > 0.05). The height and weight of the stem increased from 7 and 49 RDD. In conclusion, the harvest age affected the L. corniculatus genotypes, with 255301 being greater than the rest of the genotypes in height, leaf: stem ratio, and weight per individual stem, not in forage yield and botanical and morphological composition.

[Objectives] We evaluated the association of Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp) rectal colonisation with crude mortality and whether this association is independent of the risk of KPC-Kp infection., [Methods] This was a prospective cohort study of patients followed-up 90 days after a study of rectal colonisation. Cox regression was used to study the variables associated with crude mortality. Sensitivity analyses for 90-day crude mortality in different subcohorts were performed., [Results] A total of 1244 patients (1078 non-colonised and 166 colonised) were included. None of the non-colonised patients and 78 (47.0%) of the colonised patients developed KPC-Kp infection. The 90-day crude mortality was 18.0% (194/1078) in non-colonised patients and 41.6% (69/166) in colonised patients. Rectal colonisation was not associated with crude mortality [hazard ratio (HR) = 1.03, 95% confidence interval (CI) 0.69–1.54; P = 0.85] when the model was adjusted for severe KPC-Kp infection [INCREMENT-CPE score (ICS) > 7]. KPC-Kp infection with ICS > 7 was associated with an increased risk of all-cause mortality (HR = 2.21, 95% CI 1.35–3.63; P = 0.002). In the sensitivity analyses, KPC-Kp colonisation was not associated with mortality in any of the analysed subcohorts, including patients who did not develop KPC-Kp infection (HR = 0.93, 95% CI 0.60–1.43; P = 0.74)., [Conclusion] KPC-Kp rectal colonisation was not associated with crude mortality. Mortality increased when colonised patients developed severe KPC-Kp infection (ICS > 7). Rectal colonisation was a necessary although insufficient condition to die from a KPC-Kp infection., This work was supported by the Spanish National R&D&I Plan 2013–2016 and the Carlos III Health Institute (ISCIII), Sub-Directorate General for Cooperative Research Networks and Centers, Spanish Ministry of Economy, Industry and Competitiveness, Spanish Network for Research in Infectious Diseases [REIPI RD16/0016/0001; RD16/0016/0008] and co-financed by the European Regional Development Fund ‘A Way to Achieve Europe’, Operative Program Intelligent Growth 2014–2020.

Author Correction: Genomic surveillance of enterovirus associated with aseptic meningitis cases in southern Spain, 2015-2018. Sci Rep. 2021 Nov 18;11(1):22899. doi: 10.1038/s41598-021-02194-2. New circulating Enterovirus (EV) strains often emerge through recombination. Upsurges of recombinant non-polio enteroviruses (NPEVs) associated with neurologic manifestations such as EVA71 or Echovirus 30 (E30) are a growing public health concern in Europe. Only a few complete genomes of EVs circulating in Spain are available in public databases, making it difficult to address the emergence of recombinant EVs, understand their evolutionary relatedness and the possible implication in human disease. We have used metagenomic (untargeted) NGS to generate full-length EV genomes from CSF samples of EV-positive aseptic meningitis cases in Southern Spain between 2015 and 2018. Our analyses reveal the co-circulation of multiple Enterovirus B (EV-B) types (E6, E11, E13 and E30), including a novel E13 recombinant form. We observed a genetic turnover where emergent lineages (C1 for E6 and I [tentatively proposed in this study] for E30) replaced previous lineages circulating in Spain, some concomitant with outbreaks in other parts of Europe. Metagenomic sequencing provides an effective approach for the analysis of EV genomes directly from PCR-positive CSF samples. The detection of a novel, disease-associated, recombinant form emphasizes the importance of genomic surveillance to monitor spread and evolution of EVs. This work was funded by the Junta de Andalucía, Consejería de Salud y Familias (Project number PI-0216-2019) and by the Acción Estratégica en Salud Intramural (Project number PI20CIII/00005). MD Fernandez-Garcia received a Miguel Servet Research Contract (CP18/00067) from the Strategic Action in Health 2018 and funded by National Institute of Health Carlos III (ISCIII). ESL acknowledges funding from the INCEPTION programme (Investissements d’Avenir grant ANR-16-CONV-0005). Sí

Emiliania huxleyi is a cosmopolitan member of the marine phytoplankton. This species’ capacities for carbon sequestration and sulfur mobilization make it a key player in oceanic biogeochemical cycles that influence climate on a planetary scale. Seasonal E. huxleyi blooms are abruptly terminated by viral epidemics caused by a clade of large DNA viruses collectively known as coccolithoviruses (EhVs). EhVs thereby mediate a significant part of material and energy fluxes associated with E. huxleyi population dynamics. In this study, we use spontaneous Raman microspectroscopy to perform label-free and non-invasive measurements of the macromolecular composition of individual virions and E. huxleyi host cells. Our novel autofluorescence suppression protocol enabled spectroscopic visualization of evolving macromolecular redistributions in individual E. huxleyi cells at different stages of EhV infection. Material transfer from E. huxleyi hosts to single EhV-163 virions was confirmed by combining stable isotope probing (SIP) experiments with Raman microspectroscopy. Inheritance of the host cells’ 13C-enriched isotopic signature was quantified based on red shifts of Raman peaks characteristic of phenylalanine’s phenyl ring. Two-dimensional Raman mapping of EhV-infected E. huxleyi cells revealed that the compact region producing an intense Raman DNA signal (i.e., the nucleus) in healthy E. huxleyi cells becomes diffuse during the first hours of infection. Raman DNA emissions integrated throughout individual cells decreased during the infection cycle. Our observations are consistent with EhV-163 degrading the host’s nuclear DNA, scavenging released nucleotides for its own genome replication, and shedding newly-produced virions prior to host lysis via budding.

Background Guillain–Barré syndrome (GBS) is an acute inflammatory neuropathy with a heterogeneous presentation. Although some evidences support the role of autoantibodies in its pathogenesis, the target antigens remain unknown in a substantial proportion of GBS patients. The objective of this study is to screen for autoantibodies targeting peripheral nerve components in Guillain–Barré syndrome. Methods Autoantibody screening was performed in serum samples from all GBS patients included in the International GBS Outcome study by 11 different Spanish centres. The screening included testing for anti-ganglioside antibodies, anti-nodo/paranodal antibodies, immunocytochemistry on neuroblastoma-derived human motor neurons and murine dorsal root ganglia (DRG) neurons, and immunohistochemistry on monkey peripheral nerve sections. We analysed the staining patterns of patients and controls. The prognostic value of anti-ganglioside antibodies was also analysed. Results None of the GBS patients (n = 100) reacted against the nodo/paranodal proteins tested, and 61 (61%) were positive for, at least, one anti-ganglioside antibody. GBS sera reacted strongly against DRG neurons more frequently than controls both with IgG (6% vs 0%; p = 0.03) and IgM (11% vs 2.2%; p = 0.02) immunodetection. No differences were observed in the proportion of patients reacting against neuroblastoma-derived human motor neurons. Reactivity against monkey nerve tissue was frequently detected both in patients and controls, but specific patterns were only detected in GBS patients: IgG from 13 (13%) patients reacted strongly against Schwann cells. Finally, we confirmed that IgG anti-GM1 antibodies are associated with poorer outcomes independently of other known prognostic factors. Conclusion Our study confirms that (1) GBS patients display a heterogeneous repertoire of autoantibodies targeting nerve cells and structures; (2) gangliosides are the most frequent antigens in GBS patients and have a prognostic value; (3) further antigen-discovery experiments may elucidate other potential antigens in GBS.

Domestic pig breeds reached America on the second Columbus trip, from this date, Iberian pig genetic resources were disseminated throughout the continent, forming diverse creole breeds. These Ecuadorian Creole pigs are important for food production but have been genetically eroded since the introduction of transboundary breeds. In this study, we sought to characterize this erosion more thoroughly through mitochondrial DNA D-Loop analysis of Ecuadorian Pillareño Creole pigs from seven regions of Ecuador. To allow comparison, we also included in our analysis sequences from wild species, commercial lines, and domestic pigs, which were obtained from the NCBI GenBank database. Creole pigs’ population showed overall moderate Hd values and low π values, and a negative value of Tajima’s D was observed. The greatest differentiation from the Ecuadorian Pillareño Creole pigs was observed between Asian wild and Asian domestic pigs. The haplotype analysis revealed three different phylogenetic clades (A, E I, and E II) and 65 haplotypes. Ecuadorian Creole populations were grouped into nine haplotypes for Clade E I and E II, which have not previously been reported for Creole Pillareño populations. Our analysis indicates that in the establishment of Creole Pillareño pigs, individuals most likely separated from the Asian pig population and appear to be genetically influenced by European and Iberian populations raised in Spain.

Vitamin D has been associated with risk, development, and progression of cancer. However, the genes involved in its metabolism are highly polymorphic, compromising its activity. The aim of this study is to evaluate the association between the gene polymorphisms involved in the metabolic pathway of vitamin D and survival in patients with non-small-cell lung cancer (NSCLC). The study was designed as an observational cohort which included 194 Caucasians patients from southern Spain with NSCLC. Real-time polymerase chain reaction was used to analyze the following polymorphisms: CYP27B1 rs4646536, rs3782130, and rs10877012, CYP24A1 rs6068816 and rs4809957, GC rs7041, CYP2R1 rs10741657, VDR rs1544410 (BsmI), rs11568820 (Cdx-2), rs2228570 (FokI), rs7975232 (ApaI), and rs731236 (TaqI). Progression-free survival (PFS) and overall survival were assessed. Cox regression showed that rs4646536 was associated with PFS in the general population (p = 0.0233) and in the non-resected NSCLC subgroup (p = 0.0233). In the resected NSCLC subgroup, rs11568820 was associated with OS (p = 0.0129) and rs7041 with PFS (p = 0.0447). In the non-resected NSCLC subgroup, rs6068816 was associated with PFS (p = 0.0048) and OS (p = 0.0089) and rs731236 and rs7975232 were associated with OS (p = 0.0005) and PFS (p = 0.0002), respectively. The other polymorphisms showed no effect on the results. The rs4646536, rs6068816, rs7041, rs11568820, rs731236, and rs7975232 polymorphisms are associated with survival in NSCLC and may have a substantial role as prognostic markers of the disease.

In this study, we aimed to evaluate the genetic diversity within and among chicken breeds from the northeast region of Brazil (states of Bahia and Piau&iacute, ) using microsatellite markers. In addition, we assessed the identity and genetic relationships of chickens from Europe, Africa, and South America, as well as their influence on the formation of the Brazilian breeds. A total of 25 microsatellite markers and a panel containing 886 samples from 20 breeds (including the Brazilian chickens) were used in this study. Different statistical parameters were used to estimate the genetic diversity and relationship among the genetic groups studied. Our study indicates that the Brazilian Creole chickens have high genetic variability. The results show that chickens reared in the states of Bahia and Piau&iacute, could have originated from different ancestors. The Brazilian breeds studied have an evolutionary relationship with chickens from Portugal, Nigeria, Chile, and Spain. Our results will contribute directly to the conservation and recognition of Brazilian Creole chicken breeds and provide a solid basis for the demonstration of their genetic identity and genetic conservation of American Creole chicken populations.

Background Blood biochemistry and reference intervals help to differentiate between healthy and ill dogs as well as to provide information for the prognosis, evaluation, and monitoring; however, these intervals are often obtained from adult animals. It is essential to understand that puppies and adults are physiologically different, which justifies the need to obtain age-specific biochemical reference intervals. The aim of this research was to assess the potential effect of age, sex, body size, and their interaction on routine biochemical analytes and physiological constants (body temperature, heart rate, and respiratory rate). To carry out the research, we selected 197 healthy dogs of both sexes and different body sizes (small, medium and large) classified by age: group I (4–8 wk), group II (9–24 wk), group III (25–52 wk), and group IV (> 52 wk). The biochemical analysis included the measurement of the enzymatic activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), and the concentrations of cholesterol, triglycerides, total proteins, albumin, globulins, glucose, urea, and creatinine. Statistical analyses used analysis of variance (ANOVA) and a general linear model (GLM), which allows the comparison of multiple factors at two or more levels (p Results The results of this study showed that ALT, total protein, albumin, globulin, urea, creatinine, and body temperature levels were lower in puppies than in adult dogs of group IV (p p > 0.05). Conclusions Some biochemical components are influenced by age. For this reason, this manuscript contributes with additional data for the clinical interpretation of blood biochemical results in puppies.

In this study, tularemia outbreaks associated with humans and several domestic and wild animals (Iberian hares, wild rabbits, voles, mice, grey shrews, sheep, dogs, foxes, wolves, ticks, and river crayfish) are reported in Spain from 2007 to 2020. Special attention was paid to the outbreaks in humans in 2007–2009 and 2014–2015, when the most important waves occurred. Moreover, positive rates of tularemia in lagomorphs were detected in 2007–2010, followed by negative results in 2011–2013, before again returning to positive rates in 2014 and in 2017 and in 2019–2020. Lagomorphs role in spreading Francisella tularensis in the epidemiological chain could not be discarded. F. tularensis is described for the first time infecting the shrew Crocidura russula worldwide, and it is also reported for the first time infecting wild rabbits (Oryctolagus cuniculus) in Spain. Serological positives higher than 0.4% were seen for sheep only from 2007–2009 and again in 2019, while serological rates greater than 1% were revealed in dogs in 2007–2008 and in wild canids in 2016. F. tularensis were detected in ticks in 2009, 2014–2015, 2017, and 2019. Lastly, negative results were achieved for river crayfish and also in environmental water samples from 2007 to 2020.

Study Group on Mechanisms of Action and Resistance to Antimicrobials (GEMARA) on behalf of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC)., Klebsiella pneumoniae is an opportunistic Gram-negative pathogen that employs different strategies (resistance and persistence) to counteract antibiotic treatments. This study aimed to search for new means of combatting imipenem-resistant and persister strains of K. pneumoniae by repurposing the anticancer drug mitomycin C as an antimicrobial agent and by combining the drug and the conventional antibiotic imipenem with the lytic phage vB_KpnM-VAC13. Several clinical K. pneumoniae isolates were characterized, and an imipenem-resistant isolate (harboring OXA-245 β-lactamase) and a persister isolate were selected for study. The mitomycin C and imipenem MICs for both isolates were determined by the broth microdilution method. Time-kill curve data were obtained by optical density at 600 nm (OD600) measurement and CFU enumeration in the presence of each drug alone and with the phage. The frequency of occurrence of mutants resistant to each drug and the combinations was also calculated, and the efficacy of the combination treatments was evaluated using an in vivo infection model (Galleria mellonella). The lytic phage vB_KpnM-VAC13 and mitomycin C had synergistic effects on imipenem-resistant and persister isolates, both in vitro and in vivo. The phage-imipenem combination successfully killed the persisters but not the imipenem-resistant isolate harboring OXA-245 β-lactamase. Interestingly, the combinations decreased the emergence of in vitro resistant mutants of both isolates. Combinations of the lytic phage vB_KpnM-VAC13 with mitomycin C and imipenem were effective against the persister K. pneumoniae isolate. The lytic phage-mitomycin C combination was also effective against imipenem-resistant K. pneumoniae strains harboring OXA-245 β-lactamase., This study was funded by grants PI16/01163 and PI19/00878 awarded to M. Tomás within the State Plan for R+D+I 2013–2016 (National Plan for Scientific Research, Technological Development and Innovation 2008–2011) and cofinanced by the ISCIII-Deputy General Directorate for Evaluation and Promotion of Research–European Regional Development Fund and Instituto de Salud Carlos III FEDER, Spanish Network for the Research in Infectious Diseases (REIPI; RD16/0016/0001, RD16/0016/0006, and RD16/CIII/0004/0002) and by the Study Group on Mechanisms of Action and Resistance to Antimicrobials, GEMARA (SEIMC; http://www.seimc.org/). M. Tomás was financially supported by the Miguel Servet Research Program (SERGAS and ISCIII). I. Bleriot was financially supported by pFIS program (ISCIII, FI20/00302). O. Pacios and M. López were financially supported by grants IN606A-2020/035 and IN606B-2018/008, respectively (GAIN, Xunta de Galicia), and P. Domingo-Calap was financially supported by the ESCMID Research Grant 20200063.

Most studies focus on damage caused by beetle larvae to various crops; however, little is known about the association of adults with wild and ornamental species, their food preferences, and plant-beetle relationships. Therefore, the objective of this work was to report new host plants and floral associations for four species of Cetoniinae and Rutelinae beetles (Coleoptera: Scarabaeidae) and to provide a checklist of Scarabaeidae from San Andres Huayapam, Oaxaca, Mexico. The checklist had 15 species grouped into 12 genera, nine tribes, and four subfamilies. Twelve species had new locality records, of which one was the first accurate distribution record for the State of Oaxaca.

A set of 207 Streptococcus suis isolates were collected from ten autonomous communities from Spain in 2019 to 2020 from pigs with meningitis, pneumonic lungs, arthritic joints or other swollen viscera, to a lesser extent. Thirteen capsular types were detected being the most prevalent serotype 2 (21.7%), followed by serotypes 1 (21.3%), 9 (19.3%) and 3 (6.3%). Serotypes 2 and 9 were recovered mainly from the central nervous system (CNS), while serotype 1 was isolated mostly from swollen joints and serotype 3 from the lungs. Twenty-five isolates (12.1%) could not be typed. The most prevalent pathotype was epf + mrp + sly + luxS (49 isolates, 23.8%), and it was related mainly to serotypes 1 and 2. Serotypes 1–3 and 9 were significantly associated with anatomical sites of isolation and virulence factors, serotype 9 (CNS) and serotypes 3 and 9 (lungs) being associated with virulence profiles without the epf gene. S. suis isolates showed globally high antimicrobial resistances, but ampicillin followed by spectinomycin and tiamulin resulted in the highest activities, while the greatest resistances were detected for sulphadimethoxine, tetracyclines, neomycin, clindamycin and macrolides. A total of 87.4% isolates were positive to the tetO gene, 62.4% to the ermB gene and 25.2% to the fexA gene, while 14.6% were positive to all three genes simultaneously. A significative association between isolate resistances to tetracyclines and macrolides and the resistance genes tested was established, except for phenicol resistance and the fexA gene. A set of 14 multiresistance patterns were obtained according to the number of antimicrobials to which the isolates were resistant, the resistances to 12 or more agents being the most prevalent ones. A remarkable amount of multiresistance profiles could be seen among the S. suis serotype 9 isolates.

The dental plaque is an oral microbiome hardly associated to be the etiological agent of dental caries and periodontal disease which are still considered serious health public problems. Silver nanoparticles (AgNPs) have demonstrated to have good antimicrobial properties affecting a wide variety of microorganisms, including oral bacteria; however, there is no scientific information that has evaluated the antimicrobial effect of AgNPs against clinical oral biofilms associated with dental caries and periodontal disease. The aim of this study was to determine the antimicrobial and substantivity effects of AgNPs in oral biofilms isolated clinically from patients with dental caries and periodontal disease. Sixty-seven young and young-adult subjects with dental caries and periodontal disease were clinically sampled through the collection of subgingival dental plaque. The inhibitory effect of AgNPs was performed with standard microbiological assays by triplicate using two sizes of particle. Polymerase chain reaction (PCR) assay was used to identify the presence of specific bacterial species. All AgNPs showed an inhibitory effect for all oral biofilms for any age and, generally, any gender (p>0.05); however, the effectiveness of the antimicrobial and substantivity effects was related to particle size, time, and gender (p<0.05). The identified microorganisms wereS. mutans,S. sobrinus,S. sanguinis,S. gordonii,S. oralis,P. gingivalis,T. forsythia, andP. intermedia. The AgNPs could be considered as a potential antimicrobial agent for the control and prevention of dental caries and periodontal disease.

Reversible encapsulation of CH 2 Cl 2 or Xe in a non–porous solid–state molecular organometallic framework of [Rh(Cy 2 PCH 2 PCy 2 )(NBD)][BAr F 4 ] occurs in single–crystal to single–crystal transformations. These processes are also probed by solid–state NMR spectroscopy, including 129 Xe SSNMR. Non–covalent interactions with the –CF 3 groups, and hydrophobic channels formed, of [BAr F 4 ] – anions are shown to be important, and thus have similarly to the transport of substrates and products to and from the active site in metalloenzymes.

Severe infectious diseases are often characterized by an overwhelming and unbalanced systemic immune response to microbial infections. Human antithrombin (hAT) is a crucial coagulation inhibitor with anti-inflammatory activities. Here we identify three hAT-binding proteins (CD13, CD300f and LRP-1) on human monocytes that are involved in blocking the activity of nuclear factor-κB. We found that the modulating effect is primarily restricted to the less abundant β-isoform (hβAT) of hAT that lacks N-glycosylation at position 135. Individuals with a mutation at this position have increased production of hβAT and analysis of their blood, which was stimulated ex vivo with lipopolysaccharide, showed a decreased inflammatory response. Similar findings were recorded when heterozygotic mice expressing hAT or hβAT were challenged with lipopolysaccharide or infected with Escherichia coli bacteria. Our results finally demonstrate that in a lethal E. coli infection model, survival rates increased when mice were treated with hβAT one hour and five hours after infection. The treatment also resulted in a reduction of the inflammatory response and less severe organ damage.

Filifactor alocis and Dialister pneumosintes have been associated with the initiation and progression of periodontitis (PE). We determined and compared the frequency of both bacteria in patients with PE, rheumatoid arthritis (RA), and PE/RA simultaneously. Detection was performed by polymerase chain reaction in the subgingival biofilm. Bacteria were more frequent in patients with PE, and clinical periodontal parameters such as pocket depth (PD) and clinical attachment loss (CAL) were significantly higher in patients with PE/RA. F. alocis and D. pneumosintes could influence PD and CAL, hence participating in the initiation and progression of PE in patients with RA.

TLRs, Siglecs and CD163 are cell surface receptors that play an important role in immune response and sepsis. The objective of this study was to assess changes in the expression levels of several of these receptors (TLR2, TLR4, CD163, Siglec-1, Siglec-3, Siglec-5 and Siglec-10) on the surface of peripheral blood mononuclear cells from pigs with sepsis caused by Haemophilus parasuis. Flow cytometry was employed to analyze samples from an experimental infection and from cell cultures. A significant increase in CD163, TLR2 and Siglec-3 expression during infection was seen. However, in vitro exposure of peripheral blood monocytes to bacteria or sera from infected pigs did not increase the expression of these receptors. These changes may be due to recruitment of monocytes into the blood compartment in response to H. parasuis-induced sepsis.

The anatomical characteristics of oak wood, which is usually employed for making wine barrels, affect the properties of the wine, because these characteristics are related to the chemical compounds that can be potentially transferred to wine during the aging process. Grain, as the rate of tree growth, is the most commonly employed parameter to define the quality of the oak wood used in cooperage. Nevertheless, oxygen is a key factor in this process and there are few articles that evaluate the oxygen transmission rate (OTR) of the wood employed in cooperage. This work presents the evaluation of the anatomical characteristics of the French oak (Quercus petraea Liebl.) measured with X-ray computed tomography and the OTR of 267 wood samples extracted from recently constructed wine barrels. This study demonstrates that the OTR is dependent on the anatomical features of oak wood, the first step to classify a natural material as wood for custom oxygenation barrel making. OTR cannot be determined from the wood grain or any other single property because it depends on the overall anatomical characteristics of each wood sample, the grain and the average earlywood and latewood width being the three parameters with a stronger correlation with the OTR.

The genotoxicity of the herbicides clomazone, linuron and simazine was detected using the somatic wing spot assay of Drosophila melanogaster . For the evaluation of the mutagenic and recombinogenic activities induced by the herbicides two crosses were used: the standard cross (ST) which expresses basal levels of cytochrome P450 enzymes and the high bioactivation cross (HB) with overexpression of P450 genes conferring increased sensitivity to promutagens and procarcinogens. Third-instar larvae were exposed by chronic feeding (48 h) to three different concentrations of each herbicide. The frequencies of spots per individual in the treated series were compared to the negative concurrent control series (water or 5 % ethanol solution depending on the herbicide). Clomazone showed positive results for small and total spots in both the ST and the HB crosses. Linuron was positive at all concentrations tested for small and total spots in the ST cross while only positive for small and total spots at the highest concentration assayed in the HB cross. Simazine did not produce positive results in the ST cross while it was positive in the HB cross. The bifunctional cross-linking agent mitomycin C (MMC – 0.15 mM) was used as positive control and produced as expected a significant increase of all types of spots in both crosses. In conclusion, the positive results were due to induced mutations and not by recombinogenic activity, furthermore the significant increase in the frequency of total spots was mainly due to the induction of small single spots.

Mining activity in Cartagena‐La Union mining district (SE Spain) dates back more than 2500 years. Consequently, large amounts of sterile materials with large metal(loid) contents (mostly Zn and Pb) have accumulated in tailings ponds. A reclamation strategy based on aided phytostabilization was carried out to reclaim one tailings pond to reduce environmental and public health hazards. The main goal of this study was to determine the effectiveness of the aided phytostabilization strategy after 30 months from its implementation in enhancing soil quality and fertility and reducing mobility of the metals. Marble waste, raw pig slurry and its solid phase after physical separation were applied as amendments, and 13 Mediterranean native plant species were planted or sown. Various soil physicochemical and biochemical properties, together with the pseudo‐total and bioavailable fraction of the metal(loid)s, As, Cd, Cu, Pb and Zn, were monitored for 30 months. Results showed that pH, total organic carbon (TOC), total nitrogen (TN), cation exchange capacity (CEC), available phosphorus (P), exchangeable potassium (K), microbial biomass carbon (MBC), basal soil respiration (BSR) and enzyme activities increased with the application of the amendments and development of vegetation. In addition, the available fraction of metals markedly decreased (90–99%). Most plant species present in the area did not transfer large amounts of trace elements to the aerial tissues. The roots of Lygeum spartum loefl. Ex L. and Piptatherum miliaceum (L.) Coss. accumulated the largest amounts of As, Pb and Zn. Thus, the application of marble waste to increase pH and immobilize metal(loid)s, and pig slurry as a source of organic matter and nutrients, together with the development of vegetation to immobilize metal(loid)s in the roots and rhizosphere, and the effect of microbial communities on organic matter stabilization and development, have contributed to improve soil quality and fertility. Once the soil quality of mine tailings was improved, the development of vegetation and microbial colonization increased, promoting integrated reclamation of the area. Therefore, aided phytostabilization was a suitable technique for remediation of this tailings pond. HIGHLIGHTS: We evaluated effectiveness of mid‐term and large‐scale aided phytostabilization. Marble waste, pig slurry and native plants were used for reclamation of a tailings pond. Increased soil quality promoted plant development and microbial colonization. Lygeum spartum and Piptatherum miliaceum are good candidates to phytostabilize Pb, Zn and As.

Paroxysmal Nocturnal Hemoglobinuria (PNH) is a clonal disease of blood cells caused by the lack of glycosyl phosphatidyl inositol anchored proteins bound to the cell membrane. In consequence, erythrocytes lead to intravascular hemolysis upon complement activation, which promotes high risk of thrombosis, intravascular hemolytic anemia, and bone marrow failure in patients. The mechanisms of thrombosis in PNH are still poorly understood. Treatment with eculizumab reduces intravascular hemolysis and thrombotic risk, but not in all cases. Exosomes are extracellular vesicles released by cells and whose secretion is closely related to the inflammatory status. They participate in cell communication by activating signaling pathways and transferring genetic material and proteins to host cells. In consequence, exosomes may serve as surrogate biomarkers for the prognosis and/or diagnosis of a disease. Isolation of exosomes was carried out from healthy controls and from three groups of PNH patients, i.e. i) with no eculizumab treatment; ii) under treatment with eculizumab that have not suffered thrombosis; and iii) under treatment with eculizumab but that have suffered thrombosis. The miRNAome and proteome was analyzed using plasma focus miRNAs PCR panel and LC-MS analysis respectively. We found differential expression of miRNAs miR-148b-3p, miR-423-3p, miR29b-3p, miR15b-5p, let-7e-5p, miR126-3p, miR-125b-5p and miR-376c-3p as well as hemoglobin, haptoglobin, protein S and C4-binding protein in healthy controls vs PNH patients. Our results warrant further research and provide new information on the content of exosomes that could play a role in the hypercoagulable state in this disease.

Objective The study aims to investigate the ovarian reserve in premenopausal women with antiphospholipid syndrome (APS) and to evaluate whether it is associated with cumulative organ damage or the risk of clinical complications. Methods This single-center study was conducted in 23 premenopausal female patients (10 with primary APS and 13 with secondary APS) and 24 healthy volunteers. Serum anti-Müllerian hormone (AMH) levels were measured by enzyme-linked immunoassay. Disease-specific organ damage (DIAPS score) and the risk of clinical complications (aGAPSS score) were additionally evaluated in APS patients. Results Serum AMH levels were similar in APS patients (median 6.06, interquartile range 4.31–7.54 ng/ml) and in controls (4.87, 2.64–6.40 ng/ml; P = 0.116), and no differences were observed between the primary (6.60, 5.49–8.88 ng/ml) and secondary (6.06, 3.91–7.30 ng/ml; P = 0.532) forms of the syndrome. In individuals with APS, serum AMH levels correlated inversely with the aGAPSS score (rho–0.421, 95% confidence intervals −0.716 to −0.001; P = 0.045), while no associations were observed with the DIAPS score (rho–0.001, −0.423 to 0.422; P = 0.996). Conclusions Ovarian reserve is not reduced in premenopausal women with APS. In addition, serum AMH levels may reflect the risk of APS-related clinical complications but not the burden of disease-specific organ damage.

Resveratrol is a polyphenol with chemopreventive properties against prostate cancer; however, the mechanisms underlying its actions are not completely understood. Previously, we demonstrated that DUSP1 induces apoptosis in prostate cancer cells; therefore in the present study we investigated the role of this phosphatase on resveratrol effects. Moreover, we analysed the efficiency of combined treatment of resveratrol and the chemotherapeutic drug cisplatin on cellular viability and apoptosis and its relation with DUSP1 in prostate cancer cells. We found that resveratrol up-regulates DUSP1 expression in androgen-independent prostate cancer cells, which in turn, is involved in the inhibition of the NF-κB pathway and Cox-2 expression. This phosphatase is required for the induction of apoptosis achieved by resveratrol, but does not regulate the effects of this compound on cell cycle. Furthermore, we show that resveratrol cooperates with cisplatin both in the up-regulation of DUSP1 levels and in the promotion of apoptosis, suggesting that DUSP1 is a major determinant of cisplatin sensitivity to apoptosis. These results reveal a novel molecular mechanism by which resveratrol induces apoptosis in prostate cancer cells, and highlight the importance of DUSP1 in future therapeutic approaches based in the use of this polyphenol and cisplatin., This work was supported, in part, by Fondo de Investigaciones Sanitarias (PI070832). D.M.-M. and A.S. were recipients of grants from the “Post-Máster Program of Dpt. Biochemistry UAM”. B.G.-A. was funded by a grant from Instituto de Salud Carlos III. B.G. was funded by a grant from MINECO.