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1. Virtual emotions and Criminal Law

Author

María Isabel González-Tapia

Subjects
virtual reality, Criminal Law, emotions, metaverse-crimes, neurolaw, neurorights, Psychology, BF1-990
Abstract

This article examines the role that Criminal Law should play in regulating the non-therapeutic use of immersive Virtual Reality (VR), specifically its massive use by consumers. The starting point has been to consider VR as an intermediate risk scenario, for the purposes of Criminal Law, between the criminality entirely generated in the physical world and that developed in the 2D digital environments [cybercrimes and criminality linked to social networks and persuasive Artificial Intelligence (AI)]. Firstly, specialize literature has been analyzed to establish the nature of virtual reality. From a technical standpoint, virtual reality is a neurotechnology infused with high-risk artificial intelligence; an inseparable synthesis of non-invasive neurotechnology and a set of AI systems, considered high-risk for the fundamental rights of citizens. From the perspective of its functioning, VR is a “transformative” neurotechnology capable of altering what people perceive as reality. This is possible, because its realism lies in the emotional immersion of the user in the virtual experience, similarly to how our brain functions. Therefore, the key idea in the immersive functioning of virtual reality is its capacity to evoke and modify human emotions, which results its greater harmful potential compared to the 2D environment. From there, three central and specific areas of (legally unaddressed) risk arise: (1) the special comprehensive nature of the data collected and stored during its use; (2) its ability to mentally reproduce the “physical” experience of the avatar in the user; and (3) its significant capacity to manipulate individuals. Secondly, the paper examines both the reported cases and the foreseeable criminality in virtual worlds or “proto-metaverse,” focusing on the three risk areas, and exemplifying them with attacks on mental privacy, sexual freedom, and consumer manipulation. Finally, it is proposed that Criminal Law should also intervene (as soon as possible) to define the “red lines” of massive virtual reality use by citizens. With a democratic and human-centered approach, a basic legal framework is outlined for the criminalization of specific harms and risks associated with virtual reality, adapting the existing legal framework as necessary.

Published
2023
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2. Isolation of goat milk small extracellular vesicles by novel combined bio-physical methodology

Author

María Isabel González, Begoña Gallardo, Carlos Cerón, Elena Aguilera-Jiménez, Marta Cortes-Canteli, Héctor Peinado, Manuel Desco, and Beatriz Salinas

Subjects
goat milk, exosomes, small extracellular vesicles, isolation methods, differential ultracentrifugation, exosome characterization, Biotechnology, TP248.13-248.65
Abstract

Introduction: Goat milk is notable as a cost-effective source of exosomes, also known as small extracellular vesicles (sEVs). These nanoparticle-like structures are naturally secreted by cells and have emerged as potential diagnostic agents and drug delivery systems, also supported by their proven therapeutic effects. However, the complexity of goat milk and the lack of standardized protocols make it difficult to isolate pure sEVs. This work presents an optimized approach that combines well-established physical isolation methods with the biological treatment of milk with rennet.Methods: sEVs derived from goat milk were purified using a methodology that combines differential ultracentrifugation, rennet, and size-exclusion chromatography. This novel strategy was compared with two of the main methodologies developed for isolating extracellular vesicles from bovine and human milk by means of physico-chemical characterization of collected vesicles using Transmission Electron Microscopy, Western blot, Bradford Coomassie assay, Dynamic Light Scattering, Nanoparticle Tracking Analysis and Zeta Potential.Results: Vesicles isolated with the optimized protocol had sEV-like characteristics and high homogeneity, while samples obtained with the previous methods were highly aggregated, with significant residual protein content.Discussion: This work provides a novel biophysical methodology for isolating highly enriched goat milk sEVs samples with high stability and homogeneity, for their further evaluation in biomedical applications as diagnostic tools or drug delivery systems.

Published
2023
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3. Elevated pulse pressure and cardiovascular risk associated in Spanish population attended in primary care: IBERICAN study

Author

Ana Moyá-Amengual, Antonio Ruiz-García, Vicente Pallarés-Carratalá, Adalberto Serrano-Cumplido, Miguel Ángel Prieto-Díaz, Antonio Segura-Fragoso, Sergio Cinza-Sanjurjo, the researchers of the IBERICAN study, Alfonso Barquilla García, Ángel Díaz Rodríguez, Carlos Escobar Cervantes, Francisco Javier Alonso Moreno, Jesús Vergara Martín, Juan José Badimón, José Polo García, Luis Rodríguez Padial, Miguel Ángel Prieto, Rafael Vidal Pérez, Sergio Cinza Sanjurjo, Sònia Miravet Jiménez, Sonsoles Velilla Zancada, José Ramón Banegas, Vicente Martín Sánchez, Vicente Pallares Carratalá, Antonio Segura Fragoso y Rafael Manuel Micó Pérez, Antonio López Téllez, Jesús Vergara Martin, María De Los Ángeles Ortega Osuna, Cristóbal Prieto Cid, Mª José Hidalgo Fajardo, José Lorente Serna, Ángel Domínguez Requena, Ricardo Alberola Cañizares, Manuel Ruiz Peña, Filomena Herrero Collado, Marcela Montes Vázquez, Rafael Ángel Carrascal Garrido, María Reyes Herrera Lozano, Beatriz Ortiz Oliva, Francisco José Anguita, Carmen Pérez Ibáñez, Carlos Alberto Cabrera Rodríguez, María José Cruz Rodríguez, Sandra Bonilla Ruiz, Rocio Reina González, Salome Abad Sánchez, Inmaculada Santana Martínez, Rafael Sánchez Jordán, Juan Mª Ramos Navas-Parejo, José Manuel Ramírez Torres, José María Beltrán Poveda, María Adoración De Cruz Benayas, Carmen Fernández Gil, Jon Iñaki Esturo Alcaine, Antonio Mora Quintero, Fernando Leiva Cepas, José Luis Carrasco Martín, Emilio García Criado, Mercedes Vázquez Blanco, Isabel Mora Ortiz, Leovigildo Ginel Mendoza, Juan Carlos Aguirre Rodríguez, Esperanza María Romero Rodríguez, José Acevedo Vázquez, Juan Gabriel García Ballesteros, María De La Paz Fernández Lara, Patricia Agüera Moreno, Eduardo Paños Maturana, Juan Manuel Ignacio Expósito, Noelia Carrillo Peñas, Carmen María Abad Faya, Ana Marina Almagro Duque, Rubén Torrescusa Camisón, Paloma Menéndez Polo, Marina Peña García, Cristina López Fernández, Ascensión Estepa Torres, Miguel Gutiérrez Jansen, Esperanza Loizaga González, Lisardo García Matarín, Enrique José Gamero De Luna, Javier Benítez Rivero, María José Gómez González, Carmen Gómez Montes, Juan Carlos Rodríguez Rodríguez, Juana María González Barranco, Josefa Ramírez Vizcaíno, María Ángeles Miranda Sánchez, Eva Trillo Calvo, Concepción Bayod Calvo, Susana Larripa De La Natividad, German Grasa Lambea, Emilio Jiménez Marín, Ana Cristina Navarro Gonzalvo, Antonio Pablo Martínez Barseló, Irene Peña León, Ángel González Pérez, Liliana Mahulea, María José Pérez Martínez, Ana Piera Carbonell, Margarita Alonso Fernández, María Montserrat Rueda Cuadrado, Rodrigo Abad Rodríguez, José Miguel Álvarez Cabo, Rubén Sánchez Rodríguez, Eva María Cano Cabo, Anny Romero Secin, Miguel Ángel Prieto Díaz, Juan Jesús García Fernández, Saúl Suárez García, Fernando García Romanos, Antonia Moreno González, María Lara Amengual Sastre, Susana Martínez Palli, José Alfonso Ramón Bauza, Jose Ortiz Bolinches, Carmen Fernández Fernández, María Isabel Orlandis Vázquez, Ana Sanchís Mezquita, Fernando Unceta Aramburu, Juan Fernando Peiró Morant, Ana Moyá Amengual, Matéu Seguí-Díaz, José Félix Zuazagoitia Nubla, Ana Echevarría Ituiño, Gregorio Mediavilla Tris, María Carmen Noriega Bosch, Esther González, María Luisa Ruiz Macho, Ruth Sendino Del Olmo, Asunción Olagorta De Prado, Ana López De Viñaspre Muguerza, Jesús Iturralde Iriso, Mª Rosario Virtus Iñurrieta, Lucas Ulloa Bahamonde, Isidro Godoy García, Fernando Rubio Sevillano, María Isabel González González, Marta Pérez Souto, Raquel De León Contreras, Sara Isabel Almeida González, Irene Almería Diez, Virginia Maria Mirabal Sánchez, Francisco José Escobar Lavado, Yoel Anta Pérez, Nayra Sánchez Hernández, Juan Luis Alonso Jerez, Ricardo Koch, Nayra Ramírez Mendoza, Héctor Suárez Hernández, Francisco Jesús Morales Escobar, E Lidia Gutiérrez Fernández, Fernando Andrés Mantecón, Ana Belén García Garrido, Asunción Vélez Escalante, Luisa Alonso Rentería, Jesús Sainz Jiménez, Guillermo Pombo Alles, Esperanza Rueda Alonso, Juan Antonio Divisón Garrote, Pedro Martínez Sotodosos, Juan Antonio Vivancos Fuster, María García Palencia, José Ambrosio Torres Moraleda, Sara González Ballesteros, Ana Carmen Gil Adrados, Antonio González Cabrera, Miguel Ángel Babiano Fernández, Guillermo Rico García, Juan José Criado-Álvarez, Pilar Torres Moreno, Francisco Javier Arribas Aguirregaviria, Alicia Sahuquillo Martínez, Lourdes Maria Santos Bejar, Miguel Laborda Peralta, Raúl Piedra Castro, Carlos Santos Altozano, Lucia González-Tarrio Polo, Pedro Valiente Maresca, Reinilda Mota Santana, Noemi Elizabeth Terrero Ledesma, Noelia Garrido Espada, Gabriela Delia Rosa Zambrana Calvi, Cristina de Castro Mesa, Blanca Cordero García, Pilar Sorrius Sitges, Ana María de Santiago Nocito, César Lozano Suárez, Juan Lorenzo Gutiérrez Montero, Juan Ignacio López Gil, María Dolores Fernández Ortega, Miren Elizari Roncal, María Ascensión López Serrano, Nuria Esther Adrián De La Fuente, Belén Angulo Fdez. De Larrea, Naiara Cubelos Fernández, Guiomar Luz Ferreiro Gómez, Diana Gómez Rodríguez, Sonia María Andrés Tuñón, María Ajenjo González, Serafín De Abajo Olea, Juan José León Regueras, César Manuel Gallego Nieto, Delio Vázquez Mallada, Maria De La O Gutiérrez García, Pablo Baz Rodríguez, José Ignacio Ferradal García, Blanca Delia De Román Martínez, Ana Arconada Pérez, Omar Mahmoud Atoui, Álvaro Morán Bayón, María Teresa Armenteros Del Olmo, Francisco Javier García-Norro Herreros, Enrique Méndez Rodríguez, Diana María Narganes Pineda, Verónica Ortiz Ainaga, Milagros Sonlei Sánchez Guevara, Laura Villota Ferreiro, M Teresa Grande Grande, Francisco Vicente Martínez Gracia, Jesús Palomo del Arco, María Dolores Moriano García, Beatriz Jiménez Muñoz, Gemma Rovira Marcelino, Diana Elizabeth Fernández Valverde, Roser Rodó Bernadó, Maria Teresa Ortiz Lupiañez, Najlaa Najih, José María Diéguez Parra, Mª Rosa Benedicto Acebo, Mari Luz Bravo Vicien, Alberto Ramón León Estella, Juan Antonio Muñoz Gómez, Alicia Mostazo Muntané, Isabel Ortega Abarca, Anna Gasol Fargas, Brenda Elizabeth Riesgo Escudero, Susana Elizabeth Riesgo, Edgar Zaballos Castellvi, Celia Cols Sagarra, Marta Herranz Fernández, Josep Alins Presas, Idaira Damas Pérez, Rosa M Alcolea Garcia, Ines Monte Collado, Roberto Genique Martínez, María José Guasch Villanueva, Teresa Rama Martínez, Lucio Pinto Pena, Josefa María Panisello Royo, Inés Gil Gil, Carlos Gómez Ruiz, Rita Sahun Font, Anna Fuentes Lloveras, Alberto Calderón Montero, María Del Mar Zamora Gómez, Elena Alarcón Cebrián, Mª José Piñero Acin, Celia Pecharroman Sacristán, M Soledad Mayayo Vicente, Mª Paz Pérez Unanua, Nuria Marañón Henrich, Saray Gómez Monreal, Sonia Redondo De Pedro, Blanca Sanz Pozo, Irene Moreno Martínez, Beatriz Lopez Uriarte, Carmelina Sanz Velasco, Amaya Gárriz Aguirre, Montserrat Rivera Teijido, German Reviriego Jaén, Jose Ignacio Aza Pascual-Salcedo, Josefa Vázquez Gallego, Julia Caballer Rodilla, Aida Herrera, Ezequiel Arranz Martínez, Ana María Gómez Calvo, Paula Morán Oliva, Mª Milagros González Béjar, Julio Antonio Heras Hitos, Olga García Vallejo, Manuel De Jesús Frías Vargas, María Jesús Castillejo Boguerin, Aurora García Lerín, Miguel Ángel María Tablado, Elena Concepción García García, Leticia De Miguel Acero, Carmen Zárate Oñate, Aránzazu Barranco Apoita, María Ester Montes Belloso, Ana María Huertas Velasco, Rafael Sáez Jiménez, Julia Natividad García Pascual, María Clemencia Zuluaga Zuluaga, Mª Cruz Díez Pérez, Antonio Ruiz García, Cristina Murillo Jelsbak, Virginia Lasso Oria, Amelia González Gamarra, Elena Rodilla Rodilla, Alberto Galgo Nafría, María Mestre de Juan, Mª Carmen García Albiñana, Mª del Pilar Moreno Cano, Paula Hernanz López, Paloma Casado Pérez, Jacinto Espinosa García, José Ignacio Prieto Romo, Leandro Fernández Fernández, Javier Sierratapia, Nieves Moreno Regidor, Francisco Javier Zaballos Sánchez, Ana Moreno Moreno, Francisco Carramiñana Barrera, Juan Jose Torres Vázquez, María José Gamero Samino, Miguel Ángel De Santiago Rodríguez, Pablo Rafael Gómez Martínez, Antonio Carlos Elías Becerra, Javier Soto Olivera, Víctor Cambero, Julián Domínguez Ávila, Andrés Simón Fuentes, Jorge Manuel De Nicolás Jiménez, Dimas Igual Fraile, Guadalupe Nieto Barco, Ignacio Araujo Ramos, Mª Luz Serrano Berrocal, Francisco Buitrago Ramírez, Minerva Gallego Marcos, Félix Suarez González, Victoriano Chavero Carrasco, Francisco Guerra Peguero, Francisco Javier Sánchez Vega, Manuel Tejero Mas, Alba Palmerín Donoso, Miguel Turégano Yedro, María Beatriz Esteban Rojas, Fátima Cabezudo Moreno, Nawson Elver Quevedo Saldaña, María Del Mar García Fenés, Timotea Garrote Florencio, José María Fernández Toro, Vicente Caballero Pajares, María José Gómez Barquero, Alejandra Rey Rañal, Elena García Del Río, Enrique Nieto Pol, Julio Álvarez Fernández, Pilar Alonso Álvarez, Mª Luisa Jorge Gómez, Antonio Calvo Guerrero, Isabel Celemín Colomina, Lucia Barreiro Casal, Juana Fernández Moreno, Mª Angelines Carballal Martínez, Nabor Díaz Rodríguez, Carlos Moral Paredes, Dolores Recarey García, Francisco Javier Iglesias Mato, Antonio Fouz Ulloa, Amparo Fidalgo González, Noelia Dios Parada, Patricia Conde Sabarís, Ana Isabel Rodríguez Pérez, Ana Inés García Palacio, Víctor Julio Quesada Varela, Lidia Romero Iglesias, Ángel Lado Llerena, Carmen Lires Rodríguez, María Luisa Carretero Díaz, José Carreira Arias, José Luís Vázquez Camino, María Del Carmen Torreiro Penas, Sandra Yáñez Freire, Daniel Rey Aldana, Carlos Piñeiro Díaz, Portal González Lorenzo, José Rodríguez Campos, Rubén Blanco Rodríguez, Manuel Portela Romero, Lucía Vilela de Castro, Sonsoles María Velilla Zancada, Rafael Crespo Sabarís, Oscar Fernando Isaula Jiménez, Jesús Manuel González Puga, Jorge Antonio Benaín Ávila, Óscar Del Toro González, Vicente Llorca Bueno, Ana María Ballesteros Pérez, Domingo J Rubira López, Mª Dolores Esteve Franco, Elena Sánchez Pablo, María Teresa Palacios López, Juan Castillo Meroño, José María Lobo Martínez, Isabel María Peral Martínez, J Eduardo Carrasco Carrasco, Armando Santo González, Juan Gomáriz García, Beatriz Ríos Morata, Laura Sánchez Iñigo, Inés Sanz Pérez, Vicente Pascual Fuster, Mª Dolores Aicart Bort, Natividad Vázquez Gómez, Carlos Lluna Gasco, Teresa Amorós Barber, Pedro Antonio Medina Cano, Miguel Monteagudo Moncho, María Jesús Larré Muñoz, Raquel Navarro Hernández, Francisco José Martínez Egea, Antonio Tramontano, Marta Ferrer Royo, Belén Persiva Saura, Juan A Contreras Torres, José Mª Tirado Moliner, Alejandro Salanova Penalba, Ariadna Cucó Alberola, Fernando María Navarro I Ros, Enrique Beltrán Llicer, Ana Seoane Novás, Inmaculada Martín Valls, Gracia Verdú Mahiques, Enrique Peña Forcada, Nieves Aguilar Gómez, Francisco Javier Sanz García, María Dolores Paradís Bueso, María Eugenia Alegre Romero, Antonio Francés Camus, María Amparo Antón Peinado, Rosa Latorre Santos, Mª Asunción Palomar Marín, María Carmen Botella García, Eva Sánchez Fresquet, Pedro Sala Paños, Tomás Sánchez Ruiz, Rosa Ana Valero Valero, María Seoane Vicente, Magdalena Martin Llinares, Antonio Masiá Alegre, José Luis Llisterri Caro, Irene Lluch Verdu, Vicente Pallarés Carratalá, Francisco Valls Roca, Rafael Manuel Micó Pérez, Carmen Barceló Dupuy, Elena Benages Vicente, María José Gimeno Tortajada, Mercedes Calleja del Ser, Martín Menéndez Rodríguez, and Rosalía Victoria Carbonell Castelló

Subjects
hypertension, pulse pressure, cardiovascular risk factors, subclinical target organ damage, hypertensive cardiovascular disease, cardiovascular disease, Diseases of the circulatory (Cardiovascular) system, RC666-701
Abstract

IntroductionElevated pulse pressure (ePP) is an independent marker of cardiovascular risk (CVR) in people older than 60, and a functional marker of subclinical target organ damage (sTOD) which can predict cardiovascular events in patients with hypertension (HTN), regardless of sTOD.ObjectiveTo evaluate the prevalence of ePP in adult population seen in primary care and its association with other vascular risk factors, sTOD and with cardiovascular disease (CVD).Materials and methodsObservational multicentre study conducted in Spain (8,066 patients, 54.5% women) from the prospective cohort study IBERICAN recruited in Primary Care. Pulse pressure (PP) was defined as the difference between the systolic blood pressure (SBP) and the diastolic blood pressure (DBP) ≥60 mmHg. Adjusted (for age and sex) ePP prevalence were determined. Bivariate and multivariate analyses of the possible variables associated with ePP were carried out.ResultsThe mean of PP was 52.35 mmHg, and was significantly higher (p

Published
2023
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4. Nasal and Fecal Microbiota and Immunoprofiling of Infants With and Without RSV Bronchiolitis

Author

Claudio Alba, Marina Aparicio, Felipe González-Martínez, María Isabel González-Sánchez, Jimena Pérez-Moreno, Blanca Toledo del Castillo, Juan Miguel Rodríguez, Rosa Rodríguez-Fernández, and Leonides Fernández

Subjects
bronchiolitis, respiratory syncytial virus, immunoprofile, nasal lavage, Haemophilus, BAFF/TNFSF13B, Microbiology, QR1-502
Abstract

Bronchiolitis associated with the respiratory syncytial virus (RSV) is the leading cause of hospitalization among infants aged < 1 year. The main objective of this work was to assess the nasal and fecal microbiota and immune profiles in infants with RSV bronchiolitis, and to compare them with those of healthy infants. For this purpose, a total of 58 infants with RSV-positive bronchiolitis and 17 healthy infants (aged < 18 months) were recruited in this case-control study, which was approved by the Ethics Committee of the Hospital Gregorio Marañón. Nasal and fecal samples were obtained and submitted to bacterial microbiota analysis by 16S rDNA sequencing and to analysis of several immune factors related to inflammatory processes. Nasal samples in which Haemophilus and/or Moraxella accounted for > 20% of the total sequences were exclusively detected among infants of the bronchiolitis group. In this group, the relative abundances of Staphylococcus and Corynebacterium were significantly lower than in nasal samples from the control group while the opposite was observed for those of Haemophilus and Mannheimia. Fecal bacterial microbiota of infants with bronchiolitis was similar to that of healthy infants. Significant differences were obtained between bronchiolitis and control groups for both the frequency of detection and concentration of BAFF/TNFSF13B and sTNF.R1 in nasal samples. The concentration of BAFF/TNFSF13B was also significantly higher in fecal samples from the bronchiolitis group. In conclusion, signatures of RSV-associated bronchiolitis have been found in this study, including dominance of Haemophilus and a high concentration of BAFF/TNFSF13B, IL-8 and sTNF.R1 in nasal samples, and a high fecal concentration of BAFF/TNFSF13B.

Published
2021
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5. Bioconversion of Beet Molasses to Alpha-Galactosidase and Ethanol

Author

María-Efigenia Álvarez-Cao, María Esperanza Cerdán, Manuel Becerra, and María-Isabel González-Siso

Subjects
Microbiology (medical), Sucrose, bioconversion, Bioconversion, lcsh:QR1-502, Saccharomyces cerevisiae, Microbiology, lcsh:Microbiology, Stachyose, invertase, 03 medical and health sciences, chemistry.chemical_compound, Food science, Raffinose, Melibiose, Original Research, 030304 developmental biology, 0303 health sciences, 030306 microbiology, Fructose, Invertase, chemistry, beet molasses, alpha-galactosidase, Fermentation, ethanol
Abstract

Molasses are sub-products of the sugar industry, rich in sucrose and containing other sugars like raffinose, glucose, and fructose. Alpha-galactosidases (EC. 3.2.1.22) catalyze the hydrolysis of alpha-(1,6) bonds of galactose residues in galacto-oligosaccharides (melibiose, raffinose, and stachyose) and complex galactomannans. Alpha-galactosidases have important applications, mainly in the food industry but also in the pharmaceutical and bioenergy sectors. However, the cost of the enzyme limits the profitability of most of these applications. The use of cheap sub-products, such as molasses, as substrates for production of alpha-galactosidases, reduces the cost of the enzymes and contributes to the circular economy. Alpha-galactosidase is a specially indicated bioproduct since, at the same time, it allows to use the raffinose present in molasses. This work describes the development of a two-step system for the valuation of beet molasses, based on their use as substrate for alpha-galactosidase and bioethanol production by Saccharomyces cerevisiae. Since this yeast secretes high amounts of invertase, to avoid congest the secretory route and to facilitate alpha-galactosidase purification from the culture medium, a mutant in the SUC2 gene (encoding invertase) was constructed. After a statistical optimization of culture conditions, this mutant yielded a very high rate of molasses bioconversion to alpha-galactosidase. In the second step, the SUC2 wild type yeast strain fermented the remaining sucrose to ethanol. A procedure to recycle the yeast biomass, by using it as nitrogen source to supplement molasses, was also developed.

Published
2019
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6. Metagenomics of an Alkaline Hot Spring in Galicia (Spain): Microbial Diversity Analysis and Screening for Novel Lipolytic Enzymes

Author

María Esperanza Cerdán, Olalla López-López, María-Isabel González-Siso, and Kamila Knapik

Subjects
Microbiology (medical), Genetics, metagenomics, Thaumarchaeota, esterase, biology, Firmicutes, lcsh:QR1-502, biology.organism_classification, Microbiology, alkaline hot spring, lcsh:Microbiology, Fosmid, Chloroflexi (class), Metagenomics, Aquificae, next-generation sequencing, Proteobacteria, beta-lactamase, Original Research, Acidobacteria, biodiversity
Abstract

A fosmid library was constructed with the metagenomic DNA from the water of the Lobios hot spring (76°C, pH=8.2) located in Ourense (Spain). Metagenomic sequencing of the fosmid library allowed the assembly of 9,722 contigs ranging in size from 500 to 56,677 bp and spanning approximately 18 Mbp. 23,207 ORFs (Open Reading Frames) were predicted from the assembly. Biodiversity was explored by taxonomic classification and it revealed that bacteria were predominant, while the archaea were less abundant. The 6 most abundant bacterial phyla were Deinococcus-Thermus, Proteobacteria, Firmicutes, Acidobacteria, Aquificae and Chloroflexi. Within the archaeal superkingdom, the phylum Thaumarchaeota was predominant with the dominant species Candidatus Caldiarchaeum subterraneum. Functional classification revealed the genes associated to one-carbon metabolism as the most abundant. Both taxonomic and functional classifications showed a mixture of different microbial metabolic patterns: aerobic and anaerobic, chemoorganotrophic and chemolithotrophic, autotrophic and heterotrophic. Remarkably, the presence of genes encoding enzymes with potential biotechnological interest, such as xylanases, galactosidases, proteases and lipases, was also revealed in the metagenomic library.Functional screening of this library was subsequently done looking for genes encoding lipolytic enzymes. Six genes conferring lipolytic activity were identified and one was cloned and characterized. This gene was named LOB4Est and it was expressed in a yeast mesophilic host. LOB4Est codes for a novel esterase of family VIII, with sequence similarity to β-lactamases, but with unusual wide substrate specificity. When the enzyme was purified from the mesophilic host it showed half-life of 1 h and 43 minutes at 50°C, and maximal activity at 40°C and pH 7.5 with p-nitrophenyl-laurate as substrate. Interestingly, the enzyme retained more than 80% of maximal activity in a broad range of pH from 6.5-8.

Published
2015
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7. Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Author

Evangelia-Georgia Kostaki, Andreas Flampouris, Timokratis Karamitros, Natalia Chueca, Marta Alvarez, Paz Casas, Belen Alejos, Angelos Hatzakis, Federico Garcia, Dimitrios Paraskevis, CoRIS, Santiago Moreno, Inma Jarrín, David Dalmau, Maria Luisa Navarro, Maria Isabel González, Jose Luis Blanco, Rafael Rubio, Jose Antonio Iribarren, Félix Gutiérrez, Francesc Vidal, Juan Berenguer, Juan González, Belén Alejos, Victoria Hernando, Cristina Moreno, Carlos Iniesta, Luis Miguel Garcia Sousa, Nieves Sanz Perez, M Ángeles Muñoz-Fernández, Isabel María García-Merino, Irene Consuegra Fernández, Coral Gómez Rico, Jorge Gallego de la Fuente, Paula Palau Concejo, Joaquín Portilla, Esperanza Merino, Sergio Reus, Vicente Boix, Livia Giner, Carmen Gadea, Irene Portilla, María Pampliega, Marcos Díez, Juan Carlos Rodríguez, José Sánchez-Payá, Juan Luis Gómez, Jehovana Hernández, María Remedios Alemán, María del Mar Alonso, María Inmaculada Hernández, Felicitas Díaz-Flores, Dácil García, Ricardo Pelazas, Ana López Lirola, José Sanz Moreno, Alberto Arranz Caso, Cristina Hernández Gutiérrez, María Novella Mena, Federico Pulido, Otilia Bisbal, Asunción Hernando, Lourdes Domínguez, David Rial Crestelo, Laura Bermejo, Mireia Santacreu, José Antonio Iribarren, Julio Arrizabalaga, María José Aramburu, Xabier Camino, Francisco Rodríguez-Arrondo, Miguel Ángel von Wichmann, Lidia Pascual Tomé, Miguel Ángel Goenaga, Ma Jesús Bustinduy, Harkaitz Azkune, Maialen Ibarguren, Aitziber Lizardi, Xabier Kortajarena, Mar Masiá, Sergio Padilla, Andrés Navarro, Fernando Montolio, Catalina Robledano, Joan Gregori Colomé, Araceli Adsuar, Rafael Pascual, Marta Fernández, Elena García, José Alberto García, Xavier Barber, Juan Carlos López Bernaldo de Quirós, Isabel Gutiérrez, Margarita Ramírez, Belén Padilla, Paloma Gijón, Teresa Aldamiz-Echevarría, Francisco Tejerina, Francisco José Parras, Pascual Balsalobre, Cristina Diez, Leire Pérez Latorre, Joaquín Peraire, Consuelo Viladés, Sergio Veloso, Montserrat Vargas, Miguel López-Dupla, Montserrat Olona, Anna Rull, Esther Rodríguez-Gallego, Verónica Alba, Marta Montero Alonso, José López Aldeguer, Marino Blanes Juliá, María Tasias Pitarch, Iván Castro Hernández, Eva Calabuig Muñoz, Sandra Cuéllar Tovar, Miguel Salavert Lletí, Juan Fernández Navarro, Jose Miguel Molina, Juan González-garcia, Francisco Arnalich, José Ramón Arribas, Jose Ignacio Bernardino de la Serna, Juan Miguel Castro, Luis Escosa, Pedro Herranz, Victor Hontañón, Silvia García-Bujalance, Milagros García López-Hortelano, Alicia González-Baeza, Maria Luz Martín-Carbonero, Mario Mayoral, Maria Jose Mellado, Rafael Esteban Micán, Rocio Montejano, María Luisa Montes, Victoria Moreno, Ignacio Pérez-Valero, Berta Rodés, Talia Sainz, Elena Sendagorta, Natalia Stella Alcáriz, Eulalia Valencia, José Ramón Blanco, José Antonio Oteo, Valvanera Ibarra, Luis Metola, Mercedes Sanz, Laura Pérez-Martínez, Angels Jaén, Montse Sanmartí, Mireia Cairó, Javier Martinez-Lacasa, Pablo Velli, Roser Font, Mariona Xercavins, Noemí Alonso, Jesús Repáraz, María Gracia Ruiz de Alda, María Teresa de León Cano, Beatriz Pierola Ruiz de Galarreta, Ignacio de los Santos, Jesús Sanz Sanz, Ana Salas Aparicio, Cristina Sarriá Cepeda, Lucio Garcia-Fraile Fraile, Enrique Martín Gayo, José Luis Casado, Fernando Dronda, Ana Moreno, María Jesús Pérez Elías, Cristina Gómez Ayerbe, Carolina Gutiérrez, Nadia Madrid, Santos del Campo Terrón, Paloma Martí, Uxua Ansa, Sergio Serrano, María Jesús Vivancos, Enrique Bernal, Alfredo Cano, Antonia Alcaraz García, Joaquín Bravo Urbieta, Ángeles Muñoz, Maria Jose Alcaraz, Maria del Carmen Villalba, Federico García, José Hernández, Alejandro Peña, Leopoldo Muñoz, David Vinuesa, Clara Martinez-Montes, Fernando García, Carlos Guerrero-Beltran, Jorge Del Romero, Carmen Rodríguez, Teresa Puerta, Juan Carlos Carrió, Mar Vera, Juan Ballesteros, Oskar Ayerdi, Antonio Antela, Elena Losada, Antonio Aguilera, Melchor Riera, María Peñaranda, María Leyes, Ma Angels Ribas, Antoni A Campins, Carmen Vidal, Francisco Fanjul, Javier Murillas, Francisco Homar, Jesús Santos, Crisitina Gómez Ayerbe, Isabel Viciana, Rosario Palacios, Carmen María González, Pompeyo Viciana, Nuria Espinosa, Luis Fernando López-Cortés, Daniel Podzamczer, Elena Ferrer, Arkaitz Imaz, Juan Tiraboschi, Ana Silva, María Saumoy, Julián Olalla, Alfonso del Arco, Javier de la torre, José Luis Prada, José María García de Lomas Guerrero, Javier Pérez Stachowski, Concepción Amador, Onofre Juan Martínez, Francisco Jesús Vera, Lorena Martínez, Josefina García, Begoña Alcaraz, Amaya Jimeno, Angeles Castro Iglesias, Berta Pernas Souto, Alvaro Mena de Cea, Carlos Galera, Helena Albendin, Aurora Pérez, Asunción Iborra, Antonio Moreno, Maria Angustias Merlos, Asunción Vidal, Inés Suárez-García, Eduardo Malmierca, Patricia González-Ruano, Dolores Martín Rodrigo, Mohamed Omar Mohamed-Balghata, Juan A Pineda, Juan Macías, Miguel Thomson, Elena Delgado, Sonia Benito, and Vanessa Montero

Subjects
HIV-1, CRF02_AG, Spain, regional dispersal, spatiotemporal characteristics, Microbiology, QR1-502
Abstract

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p < 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics.

Published
2019
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8. Comparative Metagenomic Analysis of Two Hot Springs From Ourense (Northwestern Spain) and Others Worldwide

Author

María-Eugenia DeCastro, Juan-José Escuder-Rodríguez, Manuel Becerra, Esther Rodríguez-Belmonte, and María-Isabel González-Siso

Subjects
thermophiles, extreme environments, hot springs, microbial communities, metagenomics, next generation sequencing, Microbiology, QR1-502
Abstract

With their circumneutral pH and their moderate temperature (66 and 68°C, respectively), As Burgas and Muiño da Veiga are two important human-use hot springs, previously studied with traditional culture methods, but never explored with a metagenomic approach. In the present study, we have performed metagenomic sequence-based analyses to compare the taxonomic composition and functional potential of these hot springs. Proteobacteria, Deinococcus-Thermus, Firmicutes, Nitrospirae, and Aquificae are the dominant phyla in both geothermal springs, but there is a significant difference in the abundance of these phyla between As Burgas and Muiño da Veiga. Phylum Proteobacteria dominates As Burgas ecosystem while Aquificae is the most abundant phylum in Muiño da Veiga. Taxonomic and functional analyses reveal that the variability in water geochemistry might be shaping the differences in the microbial communities inhabiting these geothermal springs. The content in organic compounds of As Burgas water promotes the presence of heterotrophic populations of the genera Acidovorax and Thermus, whereas the sulfate-rich water of Muiño da Veiga favors the co-dominance of genera Sulfurihydrogenibium and Thermodesulfovibrio. Differences in ammonia concentration exert a selective pressure toward the growth of nitrogen-fixing bacteria such as Thermodesulfovibrio in Muiño da Veiga. Temperature and pH are two important factors shaping hot springs microbial communities as was determined by comparative analysis with other thermal springs.

Published
2021
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9. Metagenomics of Thermophiles with a Focus on Discovery of Novel Thermozymes

Author

María-Eugenia DeCastro, Esther Rodríguez-Belmonte, and María-Isabel González-Siso

Subjects
Metagenomics, bioinformatics, NGS, thermophiles, Thermozymes, Microbiology, QR1-502
Abstract

Microbial populations living in environments with temperatures above 50ºC (thermophiles) have been widely studied, increasing our knowledge in the composition and function of these ecological communities. Since these populations express a broad number of heat-resistant enzymes (thermozymes), they also represent an important source for novel biocatalysts that can be potentially used in industrial processes. The integrated study of the whole-community DNA from an environment, known as metagenomics, coupled with the development of next generation sequencing (NGS) technologies, has allowed the generation of large amounts of data from thermophiles. In this review, we summarize the main approaches commonly utilized for assessing the taxonomic and functional diversity of thermophiles through metagenomics, including several bioinformatics tools and some metagenome-derived methods to isolate their thermozymes.

Published
2016
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