Exploring the powerful phytoarsenal of white grape marc against bacteria and parasites causing significant diseases

Natural extracts containing high polyphenolic concentration possess antibacterial, antiparasitic and fungicidal activities. The present research characterises white grape marc, a winemaking by-product describing their physicochemical features and antimicrobial capacities. Main components of 2 different extracts generated were phenolic acids, flavan-3-ols and their gallates, and flavonols and their glycosides. As a result of this complex composition white grape marc extracts showed pronounced bioactivities with potential uses in agricultural, pharmaceutical and cosmetic industries, among others. Specifically, polyphenol compounds were extracted by using hydro-organic solvent mixtures from the by-product of Albariño white wines (Galicia, NW Spain) production. In the present work the “in vitro” antimicrobial activity of the bioactive extracts was evaluated using two different hydro-organic mixtures (HOL & HOP). The microorganisms tested included Gram positive and negative bacteria, two Apicomplexan parasite species and one Oomycota parasite. Microbial species investigated are causing agents of several human and animal diseases, such as foodborne illnesses (Bacillus cereus, Eschericha coli, Salmonella enterica), skin infections (Staphylococcus aureus), mastitis (Streptococcus uberis), parasite infections as Malaria (Plasmodium falciparum) or Toxoplasmosis (Toxoplasma gondii), and plant infections as "chestnut ink" in chestnuts or "root rot" in avocado, both diseases caused by Phytophthora cinnamomi. Both extracts verified activity against all the tested species demonstrating their potentiality to be used for the development of biocides to control a wide range of pathogenic agents; at the same time that they contribute to winemaking industry residues valorisationThis research was supported by projects GPC2017/04 (Consolidated Research Groups Program) & ED431E 2018/01 Cross-Research in Environmental Technologies (CRETUS) (Xunta de Galicia, Spain)info:eu-repo/semantics/publishedVersio

Proteomic Characterization of Antibiotic Resistance, and Production of Antimicrobial and Virulence Factors in Streptococcus Species Associated with Bovine Mastitis. Could Enzybiotics Represent Novel Therapeutic Agents Against These Pathogens?

Streptococcus spp. are major mastitis pathogens present in dairy products, which produce a variety of virulence factors that are involved in streptococcal pathogenicity. These include neuraminidase, pyrogenic exotoxin, and M protein, and in addition they might produce bacteriocins and antibiotic-resistance proteins. Unjustifiable misuse of antimicrobials has led to an increase in antibiotic-resistant bacteria present in foodstuffs. Identification of the mastitis-causing bacterial strain, as well as determining its antibiotic resistance and sensitivity is crucial for effective therapy. The present work focused on the LC–ESI–MS/MS (liquid chromatography–electrospray ionization tandem mass spectrometry) analysis of tryptic digestion peptides from mastitis-causing Streptococcus spp. isolated from milk. A total of 2706 non-redundant peptides belonging to 2510 proteins was identified and analyzed. Among them, 168 peptides were determined, representing proteins that act as virulence factors, toxins, anti-toxins, provide resistance to antibiotics that are associated with the production of lantibiotic-related compounds, or play a role in the resistance to toxic substances. Protein comparisons with the NCBI database allowed the identification of 134 peptides as specific to Streptococcus spp., while two peptides (EATGNQNISPNLTISNAQLNLEDKNK and DLWC*NM*IIAAK) were found to be species-specific to Streptococcus dysgalactiae. This proteomic repository might be useful for further studies and research work, as well as for the development of new therapeutics for the mastitis-causing Streptococcus strainsThis work was supported by the Xunta de Galicia; the European Union Social Fund (ESF); the Spanish Ministry of Economy and Competitiveness (Project AGL 2.013-48.244-R); and by the European Regional Development Fund (ERDF) (2007-2013)S

Exploring the powerful phytoarsenal of white grape marc against bacteria and parasites causing significant diseases

Natural extracts containing high polyphenolic concentration possess antibacterial, anti-parasitic and fungicidal activities. The present research characterises two extracts based on white grape marc, a winemaking by-product, describing their physicochemical features and antimicrobial capacities. The main components of these extracts are phenolic acids, flavan-3-ols and their gallates and flavonols and their glycosides. As a result of this complex composition, the extracts showed pronounced bioactivities with potential uses in agricultural, pharmaceutical and cosmetic industries. Polyphenol compounds were extracted by using hydro-organic solvent mixtures from the by-product of Albariño white wines (Galicia, NW Spain) production. The in vitro antimicrobial activity of these extracts was evaluated on Gram-positive and Gram-negative bacteria and Apicomplexan and Oomycota parasites. Microbial species investigated are causing agents of several human and animal diseases, such as foodborne illnesses (Bacillus cereus, Escherichia coli, Salmonella enterica, and Toxoplasma gondii), skin infections and/or mastitis (Staphylococcus aureus and Streptococcus uberis), malaria (Plasmodium falciparum) and plant infections as “chestnut ink” or “root rot” (Phytophthora cinnamomi). Both extracts showed activity against all the tested species, being nontoxic for the host. So, they could be used for the development of biocides to control a wide range of pathogenic agents and contribute to the enhancement of winemaking industry by-products

Application of new information and communication methodologies (ICT) for laboratory teaching in applied physics and physical chemistry

En este artículo se presenta un trabajo de innovación en metodología docente que tiene como objetivo la mejora en el aprendizaje de las enseñanzas prácticas de laboratorio de la asignatura de Física Aplicada y Fisicoquímica de la Licenciatura de Farmacia de la Universidad de Granada. La enseñanza práctica de diversas asignaturas de esta facultad tradicionalmente se ha realizado con la colaboración de los “monitores de prácticas”, los cuales se encargan de ayudar a sus compañeros en la realización de las prácticas de laboratorio, estando siempre coordinados y supervisados por un profesor. Puesto que este sistema tiene sus carencias y limitaciones, con la metodología presentada en este trabajo se pretende que desaparezca la figura del monitor de prácticas persiguiendo al mismo tiempo que el alumno sea más receptivo y muestre mayor interés. La experiencia nos demuestra que resulta fundamental la visualización de la manipulación y las operaciones necesarias para la realización de una práctica. Las nuevas tecnologías audiovisuales de la información y la comunicación (TIC) ponen al servicio de la enseñanza herramientas muy didácticas que hacen más atractivo el aprendizaje. Con tal fin, se ha elaborado para la asignatura antes mencionada un vídeo de cada práctica en formato DVD, en los que, con objeto de guiar al alumno en el trabajo experimental, se muestra la manipulación del material y reactivos empleados en cada práctica, uso de la instrumentación específica, buenas prácticas en un laboratorio y adquisición y análisis de datos. El objetivo que se persigue es que el alumno realice la práctica simultáneamente al desarrollo del vídeo, con la opción de interactuar con el vídeo y con el propio profesor presente en la sesión. En definitiva, mediante esta metodología de prácticas innovadora se pretende la captación de la atención del alumno, su motivación, promoción del trabajo autónomo y capacidad de iniciativa facilitando de esta forma su capacidad de autoaprendizaje. Todo lo comentado, contribuye a potenciar la adaptación de los actuales estudios universitarios al Espacio Europeo de Educación Superior.The aim of this innovative teaching work is the improvement in the learning process of laboratory practical sessions of the subject ‘Applied Physics and Physical Chemistry’ of the Pharmacy degree syllabuses at the University of Granada. The laboratory practical sessions of several subjects in this Faculty have been traditionally carried out with the collaboration of ‘laboratory demonstrators’, who help to their classmates in the performance of the experiments, being always coordinated and project, we expect on one hand the figure of ‘laboratory demonstrators’ to disappear, and on the other hand to achieve that the student is more receptive and shows a higher interest in the practical sessions. Our experience proves that it is very important for the student to visualize the manipulation and methodology needed for the performance of the experiments in the laboratory. The new audiovisual Information and Communication Technologies (ICT) provide very didactic tools, which make the learning more attractive. With this purpose, we have prepared a DVD movie of each experiment planned in the practical sessions of the subject. These movies show the manipulation of the material and reagents, the use of specific instrumentation, good behaviour in the laboratory, and data acquisition and analysis, with the aim of guiding the student in the experimental work. The student will perform the experiments simultaneously to the movie projection, with the possibility of interacting with the movie itself and the senior professor present during the session. In short, we expect to enhance the motivation and own-initiative of the students, facilitating their self-learning, by means of an innovative teaching methodology that contributes to promote the adaptation to the European Space for Higher Education.El proyecto ha sido financiado por la Unidad de Innovación Docente de la Universidad de Granada (Proyecto de Innovación Docente, Código Nº 08-109

MKK6 deficiency promotes cardiac dysfunction through MKK3-p38γ/δ-mTOR hyperactivation.

Stress-activated p38 kinases control a plethora of functions, and their dysregulation has been linked to the development of steatosis, obesity, immune disorders, and cancer. Therefore, they have been identified as potential targets for novel therapeutic strategies. There are four p38 family members (p38α, p38β, p38γ, and p38δ) that are activated by MKK3 and MKK6. Here, we demonstrate that lack of MKK6 reduces the lifespan in mice. Longitudinal study of cardiac function in MKK6 KO mice showed that young mice develop cardiac hypertrophy which progresses to cardiac dilatation and fibrosis with age. Mechanistically, lack of MKK6 blunts p38α activation while causing MKK3-p38γ/δ hyperphosphorylation and increased mammalian target of rapamycin (mTOR) signaling, resulting in cardiac hypertrophy. Cardiac hypertrophy in MKK6 KO mice is reverted by knocking out either p38γ or p38δ or by inhibiting the mTOR pathway with rapamycin. In conclusion, we have identified a key role for the MKK3/6-p38γ/δ pathway in the development of cardiac hypertrophy, which has important implications for the clinical use of p38α inhibitors in the long-term treatment since they might result in cardiotoxicity.We thank S Bartlett and F Chanut for English editing. We are grateful to RJ Davis, A Padmanabhan, M Costa and C López-Otín for critical reading of the manuscript. We thank Dr. RJ Davis for the MKK3 and MKK6 KO animals and Dr. Erwin F Wagner for the p38α flox mice. We thank AC Silva (ana@anasilva illustrations.com) for help with figure editing and design. This work was funded by a CNIC Intramural Project Severo Ochoa (Expediente 12–2016 IGP) to GS and JJ and PID2019-104399RB-I00 funded by MCIN/AEI/10.13039/501100011033 to GS. BGT was a fellow of FPI Severo Ochoa CNIC Program (SVP-2013-067639) and is an American Heart Association Postdoctoral Fellow (18POST34080175). RRB is a fellow of the FPU Program (FPU17/03847). The following grants provided additional funding: GS is granted by funds from European Regional Development Fund (ERDF): EFSD/Lilly European Diabetes Research Programme Dr Sabio, Fundación AECC PROYE19047SABI and Comunidad de Madrid IMMUNOTHERCAN-CM B2017/BMD-3733; US National Heart, Lung, and Blood Institute (R01 Grant HL122352), Fondos FEDER, Madrid, Spain, and Fundación Bancaria “La Caixa (project HR19/52160013); Fundación La Marató TV3: Ayudas a la investigación en enfermedades raras 2020 (LA MARATO-2020); and Instituto de Salud Carlos III to JJ. IN was funded by EFSD/Lilly grants (2017 and 2019), the CNIC IPP FP7 Marie Curie Programme (PCOFUND-2012–600396), EFSD Rising Star award (2019), JDC-2018-Incorporación (MIN/JDC1802). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovación (MCIN) and the Pro CNIC FoundationS

Growth rate and gonadal index improvement in sea urchin culture Paracentrotus lividus (Echinoida: Echinidae).

The high demand for Paracentrotus lividus (Lamarck, 1816) in countries like France, Italy, Turkey, Belgium and Japan, has raised the level of extraction of this echinoderm in recent years and has generated wear urchin populations, which take years to regenerate. The economic importance of P. lividus in Galicia, with an average annual production of 700≤000 kg and a value close to € 2 million in 2013, has led us to perform a variety of experiences in order to analyze the possibilities of the culture of this species in Galicia in raft. We have designed three experiences: 1) Effect of five diets and three densities on the growth of juvenile P. lividus obtained from the natural environment; 2) Growth of juvenile P. lividus obtained from hatchery, fed algae, and 3) Effect of different experimental diets on gonadal index of P. lividus commercial size. The experiments were performed on a USC raft situated in the estuary of Muros-Noia (Galicia) for four years (2007 - 2010), using 2 231 total urchins, from the natural environment (450), hatchery (773) and market size (1008), being designed various experimental diets that try to increase somatic and gonadal growth in sea urchins, in order to reduce the rise time of juveniles from the wild and hatchery until they reach the size commercial (55 mm diameter), or until they get a suitable size to repopulate overfished areas of the natural environment (20 mm diameter), and the growth of the gonads of sea urchins with different diets. Monthly has taken diameter of juvenile sea urchins in order to determine the effect of different diets on growth and densities of the same, as well as determining the gonadal index (GI) in sea urchins of commercial size to see the effect took different tested diets. With the data obtained estimates that, at present, it would be profitable to feed juvenile sea urchins P. lividus in raft to reach commercial size, due to the low price of this species in the market (3€ / kg). Additonally they could reach optimum size and be used to repopulate areas where natural populations are depleted. This should be combined with the fattening of gonads and extended time of commercial exploitation. Rev. Biol. Trop. 63 (Suppl. 2): 261-272. Epub 2015 June 01.La gran demanda de Paracentrotus lividus (Lamarck, 1816) en países como Francia, Italia, Turquía, Bélgica o Japón, ha elevado el nivel de extracción de este equinodermo en los últimos años que ha generado un desgaste en las poblaciones de erizos, los cuales tardan años en regenerarse. La importancia económica de P. lividus en Galicia, con una producción media anual de 700 000 kg y un valor cercano a los 2 millones de € en el año 2013, nos ha llevado a realizar diversas experiencias con el objetivo de analizar las posibilidades del cultivo en Galicia de esta especie en batea. Se han diseñado tres experiencias: 1) Efecto de cinco dietas y tres densidades sobre el crecimiento de juveniles de P. lividus obtenidos del medio natural; (2) Crecimiento de juveniles de P. lividus obtenidos en criadero, alimentados con algas, y 3) Efecto de diferentes dietas experimentales sobre el índice gonadal de P. lividus de tamaño comercial. Las experiencias se realizaron en una batea de la USC situada en la ría de Muros-Noia (Galicia), durante cuatro años (2007 - 2010), utilizando 2 231 erizos en total, procedentes del medio natural (450), de criadero (773) y de tamaño comercial (1 008), diseñándose diversas dietas experimentales que tratan de incrementar el crecimiento somático y gonadal de los erizos, con el fin de reducir el tiempo de crecimiento de los juveniles procedentes del medio natural y de criadero hasta que alcancen la talla comercial (55 mm de diámetro), o bien hasta que consigan un tamaño adecuado para repoblar las zonas sobreexplotadas del medio natural (20 mm de diámetro), así como el crecimiento de las gónadas de erizos comerciales con diferentes dietas. Mensualmente se tomó el diámetro de los erizos juveniles con el fin de determinar el efecto de las diferentes dietas y densidades sobre el crecimiento de los mismos, así como la determinación del índice gonadal (IG) en los erizos de tamaño comercial para ver el efecto de las diferentes dietas ensayadas. Con los datos obtenidos estimamos que, en la actualidad, no sería rentable alimentar erizos juveniles de P. lividus en batea hasta alcanzar la talla comercial, debido al bajo precio que tiene esta especie en el mercado (3€ / kg aproximadamente), aunque sí sería interesante mantenerlos en cultivo en batea hasta que alcancen el tamaño óptimo para repoblar zonas donde las poblaciones naturales están mermadas, combinando esto con el engorde de gónadas con alguno de los piensos diseñados que aumenta el IG y amplía el tiempo de explotación comercial de los erizos.Trabajo financiado por JACUMAR-Secretaría General de Pesca Marítima y la Consellería do Medio Rural e do Mar de la Xunta de GaliciaS

Lesion Index Titration Using Contact-Force Technology Enables Safe and Effective Radiofrequency Lesion Creation at the Root of the Aorta and Pulmonary Artery

BACKGROUND: Ablation of some myocardial substrates requires catheter-based radiofrequency delivery at the root of a great artery. We studied the safety and efficacy parameters associated with catheter-based radiofrequency delivery at the root of the aorta and pulmonary artery. METHODS: Thirty-six pigs underwent in-vivo catheter-based ablation under continuous contact-force and lesion index (power, contact-force, and time) monitoring during 60-s radiofrequency delivery with an open-irrigated tip catheter. Twenty-eight animals were allocated to groups receiving 40 W (n=9), 50 W (n=10), or 60 W (n=9) radiofrequency energy, and acute (n=22) and chronic (n=6) arterial wall damage was quantified by multiphoton microscopy in ex vivo samples. Adjacent myocardial lesions were quantified in parallel samples. The remaining 8 pigs were used to validate safety and efficacy parameters. RESULTS: Acute collagen and elastin alterations were significantly associated with radiofrequency power, although chronic assessment revealed vascular wall recovery in lesions without steam pop. The main parameters associated with steam pops were median peak temperature >42°C and impedance falls >23 ohms. Unlike other parameters, lesion index values of 9.1 units (interquartile range, 8.7-9.8) were associated with the presence of adjacent myocardial lesions in both univariate ( P=0.03) and multivariate analyses ( P=0.049; odds ratio, 1.99; 95% CI, 1.02-3.98). In the validation group, lesion index values using 40 W over a range of contact-forces correlated with the size of radiofrequency lesions (R2=0.57; P=0.03), with no angiographic or histopathologic signs of coronary artery damage. CONCLUSIONS: Lesion index values obtained during 40 W radiofrequency applications reliably monitor safe and effective lesion creation at the root of the great arteries.This study was supported by the Fundación Interhospitalaria para la Investigación Cardiovascular (FIC) and the Heart Rhythm Section of the Spanish Society of Cardiology. The Centro Nacional de Investigaciones Cardiovasculares (CNIC) is supported by the Ministry of Science, Innovation and Universities and the Pro CNIC Foundation. The CNIC is a Severo Ochoa Center of Excellence (SEV-2015- 0505). This study was supported by grants from Fondo Europeo de Desarrollo Regional (CB16/11/00458) and the Spanish Ministry of Science, Innovation and Universities (SAF2016-80324-R).S

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children