Conjugation of different immunogenic enterococcal vaccine target antigens leads to extended strain coverage

[Abstract] Enterococci have emerged as important nosocomial pathogens due to their resistance to the most commonly used antibiotics. Alternative treatments or prevention options are aimed at polysaccharides and surface-related proteins that play important roles in pathogenesis. Previously, we have shown that 2 Enterococcus faecium proteins, the secreted antigen A and the peptidyl-prolyl cis-trans isomerase, as well as the Enterococcus faecalis polysaccharide diheteroglycan, are able to induce opsonic and cross-protective antibodies. Here, we evaluate the use of glycoconjugates consisting of these proteins and an enterococcal polysaccharide to develop a vaccine with broader strain coverage. Diheteroglycan was conjugated to these 2 enterococcal proteins. Rabbit sera raised against these glycoconjugates showed Immunoglobulin G titers against the corresponding conjugate, as well as against the respective protein and carbohydrate antigens. Effective opsonophagocytic killing for the 2 sera was observed against different E. faecalis and E. faecium strains. Enzyme-linked immunosorbent assays against whole bacterial cells showed immune recognition of 22 enterococcal strains by the sera. Moreover, the sera conferred protection against E. faecalis and E. faecium strains in a mouse infection model. Our results suggest that these glycoconjugates are promising candidates for vaccine formulations with a broader coverage against these nosocomial pathogens and that the evaluated proteins are potential carrier proteins

The Siderophore Piscibactin Is a Relevant Virulence Factor for Vibrio anguillarum Favored at Low Temperatures

Vibrio anguillarum causes vibriosis, a hemorrhagic septicaemia that affects many cultured marine fish species worldwide. Two catechol siderophores, vanchrobactin and anguibactin, were previously identified in this bacterium. While vanchrobactin is a chromosomally encoded system widespread in all pathogenic and environmental strains, anguibactin is a plasmid-encoded system restricted to serotype O1 strains. In this work, we have characterized, from a serotype O2 strain producing vanchrobactin, a novel genomic island containing a cluster of genes that would encode the synthesis of piscibactin, a siderophore firstly described in the fish pathogen Photobacterium damselae subsp. piscicida. The chemical characterization of this siderophore confirmed that some strains of V. anguillarum produce piscibactin. An in silico analysis of the available genomes showed that this genomic island is present in many of the highly pathogenic V. anguillarum strains lacking the anguibactin system. The construction of single and double biosynthetic mutants for vanchrobactin and piscibactin allowed us to study the contribution of each siderophore to iron uptake, cell fitness, and virulence. Although both siderophores are simultaneously produced, piscibactin constitute a key virulence factor to infect fish, while vanchrobactin seems to have a secondary role in virulence. In addition, a transcriptional analysis of the gene cluster encoding piscibactin in V. anguillarum showed that synthesis of this siderophore is favored at low temperatures, being the transcriptional activity of the biosynthetic genes three-times higher at 18°C than at 25°C. We also show that iron levels and temperature contribute to balance the synthesis of both siderophoresThis work was supported by grants AGL2015-63740-C2-1-R and AGL2015-63740-C2-2-R (AEI/FEDER, EU) from the State Agency for Research (AEI) of Spain, and co-funded by the FEDER Programme from the European Union. The support of Xunta de Galicia (Spain) with grant GRC-2014/007 is also acknowledgedS

FrpA Is the Outer Membrane Piscibactin Transporter in Vibrio Anguillarum: Structural Elements in Synthetic Piscibactin Analogues Required for Transport

Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG[Abstract] Piscibactin (Pcb) is a labile siderophore widespread among Vibrionaceae. Its production is a major virulence factor of some fish pathogens such as Photobacterium damselae subsp. piscicida and Vibrio anguillarum. Although FrpA was previously suggested as the putative outer membrane transporter (OMT) for ferri-piscibactin, its role in piscibactin uptake was never demonstrated. In this work, we generated mutants of V. anguillarum defective in FrpA and analyzed their ability to use piscibactin as iron source. The results showed that inactivation of frpA completely disables piscibactin utilization, and the original phenotype could be restored by gene complementation, confirming that FrpA is the OMT that mediates ferri-Pcb uptake. Additionally, the ability of several Pcb thiazole analogues, with different configurations at positions 9, 10, and 13, to be internalized through FrpA, was evaluated measuring their ability to promote growth under iron deficiency of several indicator strains. The results showed that while those analogues with a thiazole ring maintain almost the same activity as Pcb, the maintenance of the hydroxyl group present in natural piscibactin configuration at position C-13 is crucial for Fe³⁺ chelation and, in consequence, for the recognition of the ferri-siderophore by the cognate OMT. All these findings allowed us to propose a Pcb analogue as a good candidate to vectorize antimicrobial compounds, through the Trojan horse strategy, to develop novel compounds against bacterial fish diseases.This work was supported by grants RTI2018-093634-B-C21/C22 (AEI/FEDER, EU), cofunded by the FEDER Programme from the European Union, and by Grant PID2019-103891RJ-100 (AEI) from the Agency for Research (AEI) of Spain. Work in University of Santiago de Compostela and University of A Coruña was also supported by grants GRC2018/018 and GRC2018/039, respectively, from Xunta de Galicia and BLUEBIOLAB (0474_BLUEBIOLAB_1_E), Programme INTERREG V A of Spain-Portugal (POCTEP). L.A. thanks Xunta de Galicia (Spain) for a predoctoral fellowship (ED481A-2019/081) co-financed by European Social Fund (ESF). Open Access funding provided thanks to the CRUE-CSIC agreement with Springer NatureXunta de Galicia; GRC2018/018Xunta de Galicia; GRC2018/039Xunta de Galicia; ED481A-2019/08

Absolute Configuration by Vibrational Circular Dichroism of Anti-inflammatory Macrolide Briarane Diterpenoids From the Gorgonian Briareum Asbestinum

[Abstract] The four new briarane diterpenoids 2-butyryloxybriarane B-3 (2), 9-acetylbriarenolide S (3), briarenolide W (4), and 12-isobriarenolide P (5), along with briarane B-3 (1) and the five known diterpenes 6–10 were isolated from the gorgonian coral Briareum asbestinum collected from the Mexican Caribbean Sea. The structures were elucidated by 1D and 2D NMR and MS measurements. Since the structure of briarane B-3 (1) was only suggested and published without any spectroscopic support, it was herein confirmed, and the supporting data are now provided. In addition, 1 provided the opportunity to explore the sensitivity of vibrational circular dichroism (VCD) to determine the configuration of a single stereogenic center in the presence of eight other stereogenic centers in a molecule possessing a highly flexible ten-member ring. A single-crystal X-ray diffraction study, in which the Flack and Hooft parameters of 1 were determined, further confirmed that briarane B-3 is (1S,2S,6S,7R,8R,9S,10S,11R,17R)-1. This paper reports for first time the use of VCD in briarane diterpenes and with the presence of chlorine atoms. Biological evaluation of seven isolated compounds evidenced a moderate anti-inflammatory activity for compounds 6 and 9 but it did not show any cytotoxic, antiviral, antibacterial, and topoisomerase inhibitory activity.This work was supported by Grants RTI2018-093634-B-C22 (AEI/FEDER, EU) from the State Agency for Research (AEI) of Spain, co-funded by the FEDER Programme from the European Union. We thank to Xunta de Galicia for the support of Grants GRC2018/039 and ED431E 2018/03 for CICA-INIBIC strategic group and BLUEBIOLAB (o474_BLUEBIOLAB_1_E, Programme INTERREG V A of Spain-Portugal (POCTEP). JR, CJ, and D.P.-P. acknowledge CESGA for the use of the computational facilities. PJ-N acknowledges partial financial support from CONACYT-Mexico Grant No. 284194. D.P.-P. received a postdoctoral fellowship from the National Council of Science and Technology (CONACYT) of Mexico. The work was also supported by UIDB/50006/2020 with funding from FCT/MCTES through national fundsXunta de Galicia; GRC2018/039Xunta de Galicia; ED431E 2018/03México. Consejo Nacional de Ciencia y Tecnología; 284194Portugal. Fundação para a Ciência e a Tecnologia; UIDB/50006/202

Modelo estratégico integral para la implementación del Sistema de Gestión de la Seguridad y Salud en el Trabajo (SG-SST) de la empresa Latinoamericana de Seguridad Eléctrica S.A.S.

El presente trabajo contiene un Modelo Estratégico Integral para la implementación del Sistema de Gestión de la Seguridad y Salud en el trabajo (SGSST) en la empresa Latinoamericana de Seguridad Eléctrica SAS., basado en los estándares mínimos de calidad de los programas de salud ocupacional de la empresa, con el fin de verificar y evaluar si estos cumplen con los procedimientos básicos que garanticen la integridad y el bienestar de los trabajadores que participan en la organización.The present work contains an Integral Strategic Model for the implementation of the Occupational Health and Safety Management System (SGSST) in the Latin American company of Electrical Safety SAS, based on the minimum quality standards of the occupational health programs of the company, in order to verify and evaluate if these comply with the basic procedures that guarantee the integrity and well-being of the workers who participate in the organization

Vocabulario de la sociedad civil, la ruralidad y los movimientos sociales en América Latina

El Vocabulario de la Sociedad Civil, la Ruralidad y los Movimientos Sociales en América Latina tiene como objetivo desarrollar vocablos relacionados con temas de gran trascendencia para la vida colectiva de la población Latinoamericana; pretende introducir a estudiantes, personas del ámbito académico y activistas en la comprensión de estas categorías de análisis. A través de la mirada de 70 especialistas que participaron en este vocabulario, es posible comprender muchos de los términos que se utilizan dentro de la investigación social y áreas relacionadas con las ciencias políticas, ambientales y rurales, a partir de una mayor explicación y detalle. Es por ello que se inserta este trabajo desde una mirada colectiva y amplia de los conceptos que se exponen. En este libro podrá encontrar las ideas de varios autores y autoras de distintas universidades, con una visión multi, inter y transdisciplinaria. El esfuerzo que se realizó para conjuntar varios términos y analizar su compleja red de interpretaciones, permitirá que este manuscrito pueda ser consultado por estudiantes, personas del ámbito científico-académico, y ciudadanía; porque contiene el estado del arte, la historia del paulatino avance de múltiples conceptos y su vigencia en el contexto actual

Colombian consensus recommendations for diagnosis, management and treatment of the infection by SARS-COV-2/ COVID-19 in health care facilities - Recommendations from expert´s group based and informed on evidence

La Asociación Colombiana de Infectología (ACIN) y el Instituto de Evaluación de Nuevas Tecnologías de la Salud (IETS) conformó un grupo de trabajo para desarrollar recomendaciones informadas y basadas en evidencia, por consenso de expertos para la atención, diagnóstico y manejo de casos de Covid 19. Estas guías son dirigidas al personal de salud y buscar dar recomendaciones en los ámbitos de la atención en salud de los casos de Covid-19, en el contexto nacional de Colombia

Búsqueda de compuestos bioactivos a partir de bacterias del Phylumfirmicutes aisladas del octocoral pseudopterogorgia elisabethae recolectado en la Isla de Providencia

En este trabajo se emplearon 15 cepas bacterianas del Phylum Firmicutes con el fin de seleccionar una cepa ellas como productora de compuestos con actividad IQS. Para esto se llevó a cabo su cultivo en 4 medios diferentes (LBS, TAK, BM y MM), y luego se obtuvieron sus extractos mediantes el uso de la resina no iónica Diaion® HP-20, estos extractos fueron evaluados en el ensayo de actividad IQS frente al biosensor Chromobacterium violaceum ATCC 31532 (se evaluaron 64 extractos a 2.0, 1.0 y 0.2 mg/sensidisco haciendo los experimentos por duplicado). Los resultados de actividad IQS obtenidos permitieron seleccionar a las cepas Jeotgalicoccus halophilus, Bacillus licheniformis, Staphylococcus warneri, Ocenobacillus profundis, Bacillus anthracis y Bacillus horneckiae para continuar con el trabajo por tener extractos bioactivos. Luego, cada uno de los extractos bioactivos fue fraccionado por medio de CC en fase reversa obteniendo 3 fracciones (MeOH:H2O 1:9, MeOH:H2O 1:1 y MeOH), las cuales fueron evaluadas en el ensayo de actividad IQS, de esta manera se pudo establecer que la actividad IQS en todos los casos se presenta en las fracción metanólica, lo que muestra un carácter desde poco hasta medianamente polar de los compuestos responsables de la actividad IQS. En esta segunda etapa se seleccionaron las cepas J. halophilus y O. profundis para continuar con el estudio, por proporcionar las fracciones metanólica más activas y por no existir reportes de su química en la literatura. A continuación las bacterias seleccionadas J. halophilus y O. profundis fueron cultivadas en los medios LBS, TAK y BM, y sus medios agotados (100 mL x 10) fueron extraídos con DCM mediante una metodología L-L contínua lo que permitió obtener el extracto orgánico; así mismo, la fase acuosa resultante fue extraída por medio de resina Diaion® HP-20 obteniendo el extracto HP-20 residual por elución con metanol. Cada uno de estos dos extractos fue evaluado en el ensayo IQS encontrando que el extracto con mayor actividad correspondió al extracto orgánico obtenido del cultivo de J. halophilus en el medio LBS, por lo que esta cepa fue la selecionada para escalar su cultivo y obtener los compuestos bioactivos en cantidad suficiente para su identificación. En el paso siguiente se estudió cómo algunas variables en las condiciones de cultivo (volumen, agitación y tiempo de incubación) de J. halophilus en LBS, influyen en la cantidad y calidad del extracto obtenido (actividad IQS y complejidad). Encontrando que cuando se emplea agitación a 100 rpm, temperatura ambiente (~ 19 °C), durante un tiempo entre 96-144 h y haciendo cultivos de 100 mL en fiolas de 500 mL se logra obtener una buena cantidad de extracto activo. El proceso se repitió hasta obtener una masa de extracto suficiente (273 cultivos). Así mismo se estableció que el mejor método de extracción es el uso DCM en una extracción continua L-L. Una vez obtenido el extracto biactivo, se buscó establecer la influencia de la fase estacionaría en la separación de este extracto (RP-18, sílica y diol) y se pudo determinar que se obtienen los mejores resultados con resinas diol. Así, el fraccionamiento de todo el extracto se hizo por CC flash diol y se llevó a cabo de manera bioguiada, encontrando que la mayor actividad residía en las fracciones F2-F6. Estas fracciones fueron sometidas a cromatografía de exclusión de tamaño, cromatografía en columna (sílica gel) y cromatografía en placa delgada preparativa, obteniendo los siguientes compuestos con actividad IQS (en paréntesis se da la mínima concentración en la que detectamos actividad IQS): 4-(metiltio)fenol [1] (0.36 M), p-hidroxibenzaldehído [2] (0.41 M), 3-indolaldehído [3] (0.68 M), (1H-indol-3-il)oxoacetamida [4] (1.06 M), y 2-metilpropanoamida [5] (1.14 M). Todos los compuestos fueron determinados inequívocamente mediante el estudio de sus espectros de RMN y masas en modo IE, siendo los compuestos 3-4 los mayoritarios. Los compuestos 1 y 4 se reportan por primera vez para bacterias. Finalmente, fue estudiado el efecto del medio de cultivo en la producción de los compuestos bioactivos antes identificados 1-5. Para esto J. halophilus se cultivó en MOLP, KAL y LBS, con y sin adición de agar (0.3 g /L), encontrando que en el medio MOLP no hay evidencia de la producción de los compuestos indólicos 3-4, y no parece que se indujera la producción de ningún compuesto a juzgar por los análisis de CLAE y RMN, en las condiciones de cultivo probadas. Mientras, en los cultivos en LBS y KAL sí hay producción de derivados indólicos, y sorprendentmente la adición de agar en el medio LBS estimula la producción del ácido tiobenzoico [6], que nunca había sido reportado como metabolito para una bacteria. Con este estudio se contribuyó por primera vez en Colombia al desarrollo de una metodología automatizable para la selección de microorganismos capaces de producir compuestos con actividad IQS, a partir de aislamientos nativos de bacterias marinas. Lo anterior ha ayudado a abrir este campo del conocimiento en los productos naturales, del que se esperan los mayores desarrollos hacia el futuro, tanto a nivel mundial como local, y del que apenas se dan los primeros pasos en Colombia.Abstract. Fifteen bacterial strains belonged to the Phylum Firmicutes were used to develop a quorum sensing Inhibitors (QSI) screening. Each bacterium were cultured in four different media (LBS, TAK, BM y MM) and then, the compounds produced were extracted using the non-polar resin Diaion® HP-20. All the extracts (60 bacterial extracts and 4 control media) were tested using a QSI assay, against Chromobacterium violaceum ATCC 31532, at three different concentrations (2.0, 1.0 and 0.2 mg/disc) by duplicated. The culture extracts of Jeotgalicoccus halophilus, Bacillus licheniformis, Staphylococcus warneri, Ocenobacillus profundis, Bacillus anthracis and Bacillus horneckiae were selected as the most active in this preliminary screening. Subsequently each one of the extracts selected were fractionated by reverse phase column chromatography, obtaining three fractions (MeOH:H2O 1:9, MeOH:H2O 1:1 and MeOH). Each fractions were tested in the bioassay, showing in all the cases that the MeOH fraction was the most active one. Considering, the activity, the extracts yield, complexity by HPLC, the bacterial strains J. halophilus and O. profundis were selected to continue the study. Likewise, there are no reports about their chemistry. J. halophilus and O. profundis were cultured (100 mL x10) in LBS, TAK and BM culture media and the culture supernatants were extracted using a continuous liquid-liquid extraction with dichloromethane (organic extract). The remained aqueous phases were further extracted using the non-polar resin Diaion® HP-20 and named as HP-20 residual extracts. Each extract (organic and HP-20) were evaluated in the QSI assay. The results showed that DCM extract of culture of J. halophilus in LBS media was the most active. In order to know if there are changes in the yield and composition (QSI activity and complexity) of the extract of J. halophilus in LBS broth, other culture parameters as volume, shake and time were studied. Thus, J. halophilus were culture in LBS using three different volumes (100, 500 and 1000 mL), at 0 and 100 rpm, and cultured during 96 h and 144 h. Finally, the culture of J. halophilus was scaled up (100 mL X 273) at 100 rpm and 19 °C. Additionally, it was established the continous L-L extraction with DCM, as the best method for obtaining a clean extract. Subsequently, the influence of the stationary phase in the separation of the extract was studied using different phases. The best results were obtained using Diol flash CC. The bioguided fractionation demonstrated that the fractions F2 to F6 were the most active fractions. These fractions were purified by size exclusion chromatography, column chromatography and preparative thin layer chromatography yield the QSI compounds: 4- (methylthio)phenol 1 (0.36 M), p-hydroxibenzaldehide 2 (0.41 M), 3-indolaldehide 3 (0.68 M), (1H-indol-3-yl)oxoacetamide 4 (1.06 M) y 2-methylpropanoamide 5 (1.14 M), and the thiobenzoic acid 6 (activity was not determined). All of these compounds were unequivocally identified by NMR and MS-EI. The major components produced by J. halophilus were 3 and 4. The compound 1, 4 and 6 are reported by first time from bacterial source. Finally, the effect of the culture media over the production of the identified bioactive compounds was made. J. halophilus was cultured in MOLP, KAL and LBS with and without agar addition (0.3 g/L), finding that was not production of indolic compounds (3-4) or different kind of compounds, using MOLP media, judging by its HPLC-PDA and NMR analysis. While, indolic compounds were produced in KAL and LBS media, also the agar addition stimulates the production of thiobenzoic acid 6. This is the first chemical study reported for the bacterium J. halophilus and also in Colombia, this is the first report of the development of a new automatized methodology for screening of microorganisms (native marine bacteria) as producers of QSI compounds. This study allows opening a new field of knowledge in natural products from native marine microorganism. Future global and local studies will be necessary in this field of research.Maestrí

Voces de Cuba

Curs 2016-2017El 25 de novembre de 2016 va morir Fidel Castro, líder històric de la Revolució cubana. Un mes després, tres estudiants de la Universitat de Vic – Universitat Central de Catalunya, van viatjar a l’Havana per conèixer de primera mà com els cubans estaven vivint aquell fet transcendental. De les opinions i reflexions recollides, en va sortir el documental “Voces de Cuba”. Aquest document és un recull de tot allò que ha comportat realitzar aquest documental, tant en l'àmbit tècnic i logístic, com en el personal. L'objectiu és explicar quines són les decisions que l'equip ha pres i com ha estat aquest llarg procés. També hi ha una descripció detallada de la narrativa, la direcció, producció i edició, que analitza i justifica el projecte. És lògic pensar que el resultat del documental podria haver estat diferent, i hi estem d'acord. Només hauria calgut que s'incorporés algú nou a l'equip, que algú no hi hagués estat o que els factors externs s'haguessin alineat d'una altra manera. Però quan es treballa amb la realitat, i no amb la ficció, és molt difícil controlar el què i el com.On November 25th, 2016, Fidel Castro, the historical leader of the Cuban Revolution died. A month later, three students from the University of Vic – Central University of Catalonia, traveled to La Habana to learn from first person how cubans are dealing with that transcendental fact. The documentary “Voces de Cuba”, comes from a deep thinking of the situation that this country is living in this century. However, this docuement explains why “Voces de Cuba” has been made and how. Adding a detailed description of the narrative, direction of the Project, producing, editing, analyzing, and justifying this documentary. Anyone could say this project could have another prespective and this team agrees that it could have been made differently. But when working with reality, and not fiction, is hard to control what to do and how to do it after living the expereince