Diagrames Pressió-Temperatura de compostos moleculars amb activitat terapèutica

L’objectiu d’aquest projecte és la determinació de diagrames d’equilibri Pressió-Temperatura de compostos que presenten activitat terapèutica utilitzats en la indústria farmacèutica. També s’inclou en el projecte l’estudi del polimorfisme, capacitat d’una substància a existir en diferents fases sòlides, i l’establiment de l’ordre d’estabilitat de les fases. Per a l’obtenció dels diagrames s’utilitzen les tècniques d’anàlisi tèrmica diferencial, aquestes permeten caracteritzar transformacions termodinàmiques, principalment canvis d’estat, tant a pressió atmosfèrica com a altres pressions. Amb aquestes tècniques es pot conèixer la temperatura a la qual es dóna la transició de fase, així com altres dades de caire energètic. També s’han realitzat anàlisis de difracció de raig X, per una banda determinar el volum específic del compost analitzat i, per altra banda, per si aquest presenta cap transició a fases més estables. Els compostos analitzats són l’enantiòmer (R) de l’àcid 2-CloroMandelic, el compost racèmic l’àcid 2-CloroMandelic i una mescla eutèctica de composició X=0,56. Els resultats experimentals mostren com ni l’enantiòmer ni la mescla eutèctica presenten cap forma polimorfa, només el racemat en presenta. També s’ha observat com en els segons escalfament de la mescla eutèctica i el racemat tendeixen al mateix compost, el conglomerat. El treball aquí exposat té repercussió en l’àmbit farmacèutic, ja que l’estudi del polimorfisme és molt important en la formulació de fàrmacs, i també perquè aquests composts són de naturalesa quiral i, per tant, els descobriments en aquests tipus de productes poden suposar un gran avanç

Carcinoide bronquial y síndrome de neoplasias endocrinas múltiples tipo 1: aportación de un caso

Case report[Abstract] Carcinoid tumours of bronchial origin are rare in type 1 multiple endocrine neoplasia (MEN1) syndrome. The prevalence of histologically confirmed cases is approximately 5-8%, although in more recent studies it is estimated that it could be much higher and a possible relationship with the presence of hypergastrinaemia is suggested. We report a patient with a type 1 MEN syndrome, with no respiratory symptoms, with hypergastrinaemia, and in whom a 5mm diameter nodule was detected in the wall of the left main bronchus by computed tomography. The bronchial biopsy confirmed that it was a typical bronchial carcinoid and the octreoscan showed a single focus of high uptake coinciding with this lesion. A bronchoplastic (sleeve) was performed with extirpation of 3 bronchial rings, which also demonstrated that it was a typical carcinoid.[Resumen] Los tumores carcinoides de origen bronquial son raros en el síndrome de neoplasias endocrinas múltiples tipo 1 (MEN1). La prevalencia de casos con confirmación histológica es aproximadamente del 5%–8%, aunque en estudios más recientes se estima que podría ser más alta y, en ellos, se contempla una posible relación con la presencia de hipergastrinemía. Aportamos una paciente con síndrome de MEN1, sin síntomas respiratorios, con hipergastrinemia, y que en una tomografía computarizada se detectó un nodulo de 5 mm de diámetro en la pared del bronquio principal izquierdo. La biopsia bronquial confirmó que se trataba de un carcinoide bronquial típico y el Octeoscran® mostró un único foco de hipercaptación coincidente con esta lesión. Se realizó una broncotomía con extirpación de 3 anillos bronquiales, que también demostró que se trataba de un carcinoide típico

Protocol for cryopreservation of the turbot parasite Philasterides dicentrarchi (Ciliophora, Scuticociliatia)

This is the accepted manuscript of the following article: Folgueira, I., de Felipe, A.P., Sueiro, R.A., Lamas, J. & Leiro, J. (2018). Protocol for cryopreservation of the turbot parasite Philasterides dicentrarchi (Ciliophora, Scuticociliatia). Cryobiology, 80, 77-83. doi: 10.1016/j.cryobiol.2017.11.010Philasterides dicentrarchi is a free-living marine ciliate that can become an endoparasite that causes a severe disease called scuticociliatosis in cultured fish. Long-term maintenance of this scuticociliate in the laboratory is currently only possible by subculture, with periodic passage in fish to maintain the virulence of the isolates. In this study, we developed and optimized a cryopreservation protocol similar to that used for the long-term storage of scuticociliates of the genus Miamiensis. The cryogenic medium comprised ATCC medium 1651 and a combination of 11% dimethylsulfoxide and 5% glycerol. We have verified that the most important factor ensuring the efficiency of the cryopreservation procedure is the growth phase of the culture, and that ciliates should be cryopreserved at the stationary phase (around the sixth day of culture). The cryopreservation protocol described here can be used for all strains of P. dicentrarchi as well as commercial strains of Miamiensis and enables the virulence of the strains to be maintained. Finally, this cryopreservation protocol has been shown to be more effective than others routinely applied to scuticociliates, yielding a higher survival rate with a lower initial concentration of ciliates. The results obtained indicate that the cropreservation protocol enables the long-term storage of scuticociliate parasites while maintaining the virulence of the isolates. The protocol is therefore suitable for use in vaccine production and related studiesThis work was financially supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429 (PARAFISHCONTROL), by the Ministerio de Economía y Competitividad (Spain) under grant agreement AGL2014-57125-R and by grant GPC2014/069 from the Xunta de Galicia (Spain)S

Identification and Molecular Characterization of Superoxide Dismutases Isolated From A Scuticociliate Parasite: Physiological Role in Oxidative Stress

Philasterides dicentrarchi is a free-living microaerophilic scuticociliate that can become a facultative parasite and cause a serious parasitic disease in farmed fsh. Both the free-living and parasitic forms of this scuticociliate are exposed to oxidative stress associated with environmental factors and the host immune system. The reactive oxygen species (ROS) generated by the host are neutralized by the ciliate by means of antioxidant defences. In this study we aimed to identify metalloenzymes with superoxide dismutase (SOD) activity apable of inactivating the superoxide anion (•O2−) generated during induction of oxidative stress. P. dicentrarchi possesses the three characteristic types of SOD isoenzymes in eukaryotes: copper/zinc-SOD, manganese-SOD and iron-SOD. The Cu/Zn-SOD isoenzymes comprise three types of homodimeric proteins (CSD1-3) of molecular weight (MW) 34–44kDa and with very diferent AA sequences. All Cu/Zn-SODs are sensitive to NaCN, located in the cytosol and in the alveolar sacs, and one of them (CSD2) is extracellular. Mn- and Fe-SOD transcripts encode homodimeric proteins (MSD and FSD, respectively) in their native state: a) MSD (MW 50kDa) is insensitive to H2O2 and NaN3 and is located in the mitochondria; and b) FSD (MW 60kDa) is sensitive to H2O2, NaN3 and the polyphenol trans-resveratrol and is located extracellularly. Expression of SOD isoenzymes increases when •O2 − is induced by ultraviolet (UV) irradiation, and the increase is proportional to the dose of energy applied, indicating that these enzymes are actively involved in cellular protection against oxidative stressThis study was financially supported by grant AGL2017-83577-R awarded by the Ministerio de Economía y Competitividad (Spain) and Fondo Europeo de Desarrollo Regional -FEDER- (European Union), by grant ED431C2017/31 from the Xunta de Galicia (Spain), and by PARAFISHCONTROL project, which received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429S

New data on flatfish scuticociliatosis reveal that Miamiensis avidus and Philasterides dicentrarchi are different species

This article has been published in a revised form in Parasitology [http://doi.org/10.1017/S0031182017000749]. This version is free to view and download for private research and study only. Not for re-distribution, re-sale or use in derivative works. © 2017 Cambridge University PressScuticociliatosis is a severe disease in farmed flatfish. However, the causative agent is not always accurately identified. In this study, we identified two isolates of scuticociliates from an outbreak in cultured fine flounder Paralichthys adspersus. Scuticociliate identification was based on morphological data, examination of life stages and the use of molecular approaches. The isolates were compared with a strain of Philasterides dicentrachi from turbot Scophthalmus maximus and with a strain deposited in the American Type Culture Collection as Miamiensis avidus ATCC® 50180™. The use of morphological, biological and molecular methods enabled us to identify the isolates from the fine flouder as P. dicentrarchi. Comparison of P. dicentrachi isolates and M. avidus revealed some differences in the buccal apparatus. Unlike P. dicentrarchi, M. avidus has a life cycle with three forms: macrostomes (capable of feeding on P. dicentrarchi), microstomes and tomites. Additionally, we found differences in the 18S rRNA and α- and β-tubulin gene sequences, indicating that P. dicentrarchi and M. avidus are different species. We therefore reject the synonymy/conspecificity of the two taxa previously suggested. Finally, we suggest that a combination of morphological, biological, molecular (by multigene analysis) and serological techniques could improve the identification of scuticociliates parasites in fishThis work was financially supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429 (PARAFISHCONTROL), by the Ministerio de Economía y Competitividad (Spain) under grant agreement AGL2014-57125-R and by grant GPC2014/069 from the Xunta de Galicia (Spain)S

Evidence for the role of extrusomes in evading attack by the host immune system in a scuticociliate parasite

This is the accepted manuscript of the following article: Folgueira I, Lamas J, De Felipe AP, Sueiro RA, Leiro JM. (2019). Evidence for the role of extrusomes in evading attack by the host immune system in a scuticociliate parasite. Fish Shellfish Immunol. 2019 Jul 5;92:802-812. doi: 10.1016/j.fsi.2019.07.008Like other ciliates, Philasterides dicentrarchi, the scuticociliate parasite of turbot, produces a feeding-only or growing stage called a trophont during its life cycle. Exposure of the trophonts to heat-inactivated serum extracted from the turbot host and containing specific antibodies that induce agglutination/immobilization leads to the production of a mucoid capsule from which the trophonts later emerge. We investigated how these capsules are generated, observing that the mechanism was associated with the process of exocytosis involved in the release of a matrix material from the extrusomes. The extruded material contains mucin-like glycoproteins that were deposited on the surface of the cell and whose expression increased with time of exposure to the heat-inactivated immune serum, at both protein expression and gene expression levels. Stimulation of the trophonts with the immune serum also caused an increase in discharge of the intracellular storage compartments of calcium necessary for the exocytosis processes in the extrusomes. The results obtained suggest that P. dicentrarchi uses the extrusion mechanism to generate a physical barrier protecting the ciliate from attack by soluble factors of the host immune system. Data on the proteins involved and the potential development of molecules that interfere with this exocytic process could contribute to improving the prevention and control of scuticociliatosis in turbotThis study was financially supported by grants from the Ministerio de Economía y Competitividad and Fondo Europeo de Desarrollo Regional -FEDER- (European Union) (AGL2017-83577-R) and from the Xunta de Galicia (ED431C2017/31) and also by the PARAFISHCONTROL project, which received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No. 634429S

Eradication of pseudomonas aeruginosa with inhaled colistin in adults with non-cystic fibrosis bronchiectasis

[Abstract] The persistent isolation of Pseudomonas aeruginosa in the airways of non-cystic fibrosis bronchiectasis (NCFB) patients is associated with a worsening of the symptoms, increase of exacerbations, poor quality of life and functional impairment. The objective of this study was the analysis of the eradication rate of P. aeruginosa in the sputum of patients with NCFB treated with inhaled colistin and the effects of the treatment in the exacerbations. This was a prospective, cohort, study of 67 NCFB patients treated with inhaled colistin at the Hospital of A Corun˜a (Spain).We recorded dyspnoea, exacerbations, lung function and sputum cultures of P. aeruginosa in the patients. The mean age of the patients was 67.25+14.6 years (59.7% male). The percentages of eradication of P. aeruginosa in sputum at 3, 6, 9 and 12 months were 61.2%, 50.7%, 43.3% and 40.3%, respectively.We observed a significant decrease in exacerbations after 1 year of colistin treatment (1.98+3.62) versus the previous year (3.40 + 4.21, p < 0.001). We conclude that treatment with inhaled colistin in patients with NCFB and P. aeruginosa in sputum can achieve high rates of eradication even in patients with several previous positive cultures, as well as a significant decrease of exacerbations and hospital admissions

Combined antiparasitic and anti‐inflammatory effects of the natural polyphenol curcumin on turbot scuticociliatosis

This is the peer reviewed version of the following article: Mallo, N. , DeFelipe, A. P., Folgueira, I. , Sueiro, R. A., Lamas, J. and Leiro, J. M. (2017), Combined antiparasitic and anti‐inflammatory effects of the natural polyphenol curcumin on turbot scuticociliatosis. J Fish Dis, 40: 205217. doi:10.1111/jfd.12503, which has been published in final form at https://doi.org/10.1111/jfd.12503. This article may be used for noncommercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived VersionsThe histiophagous scuticociliate Philasterides dicentrarchi is the aetiological agent of scuticociliatosis, a parasitic disease of farmed turbot. Curcumin, a polyphenol from Curcuma longa (turmeric), is known to have antioxidant and anti-inflammatory properties. We investigated the in vitro effects of curcumin on the growth of P. dicentrarchi and on the production of pro-inflammatory cytokines in turbot leucocytes activated by parasite cysteine proteases. At 100 μm, curcumin had a cytotoxic effect and completely inhibited the growth of the parasite. At 50 μm, curcumin inhibited the protease activity of the parasite and expression of genes encoding two virulence-associated proteases: leishmanolysin-like peptidase and cathepsin L-like. At concentrations between 25 and 50 μm, curcumin inhibited the expression of S-adenosyl-L-homocysteine hydrolase, an enzyme involved in the biosynthesis of the amino acids methionine and cysteine. At 100 μm, curcumin inhibited the expression of the cytokines tumour necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) produced in turbot leucocytes activated by parasite proteases. Results show that curcumin has a dual effect on scuticociliatosis: an antiparasitic effect on the catabolism and anabolism of ciliate proteins, and an anti-inflammatory effect that inhibits the production of proinflammatory cytokines in the host. The present findings suggest the potential usefulness of this polyphenol in treating scuticociliatosisThis work was financially supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429 (PARAFISHCONTROL) and by the Ministerio de Economía y Competitividad (Spain) under grant agreement AGL2014-57125-RS

Restauración de obras escultóricas mediante el empleo de Software libre

La digitalización del patrimonio cultural es hoy en día uno de los ámbitos de las denominadas humanidades di- gitales que se encuentra en mayor expansión. Tanto con fines de investigación como para la restauración, con- servación o difusión de bienes culturales, son innumerables los contextos en los que esta disciplina resulta de utilidad. Sin embargo suele percibirse que este tipo de tareas exigen un importante desembolso económico en equipamiento y software informático, quedando su uso al alcance únicamente de grandes instituciones. Con el presente proyecto hemos intentado poner en cuestión esta creencia desarrollando un proyecto completo de res- tauración escultórica mediante únicamente software y hardware libre o gratuito. Desde el inicio del proyecto, con la creación de una mesa rotatoria automatizada para facilitar el escaneado, hasta la impresión final de las piezas faltantes en la escultura, pasando por la captura de las imágenes, la restitución fotogramétrica, el procesa- do del modelo y la creación de los fragmentos perdidos, todo ello ha sido realizado con la doble premisa de in- currir en un coste económico próximo a cero y en que la calidad y precisión de los resultados sea completamente profesional. La presente memoria relata la experiencia y permite constatar el éxito de nuestra propuesta

The coagulation system helps control infection caused by the ciliate parasite Philasterides dicentrarchi in the turbot Scophthalmus maximus (L.)

This is the accepted manuscript of the following article: Blanco-Abad, V., Noia, M., Valle, A., Fontenla, F., Folgueira, I., & De Felipe, A. et al. (2018). The coagulation system helps control infection caused by the ciliate parasite Philasterides dicentrarchi in the turbot Scophthalmus maximus (L.). Developmental & Comparative Immunology, 87, 147-156. doi: 10.1016/j.dci.2018.06.001Many studies have shown that coagulation systems play an important role in the defence against pathogens in invertebrates and vertebrates. In vertebrates, particularly in mammals, it has been established that the coagulation system participates in the entrapment of pathogens and activation of the early immune response. However, functional studies investigating the importance of the fish coagulation system in host defence against pathogens are scarce. In the present study, injection of turbot (Scopthalamus maximus) with the pathogenic ciliate Philasterides dicentrarchi led to the formation of macroscopic intraperitoneal clots in the fish. The clots contained abundant, immobilized ciliates, many of which were lysed. We demonstrated that the plasma clots immobilize and kill the ciliates in vitro. To test the importance of plasma clotting in ciliate killing, we inhibited the process by adding a tetrapeptide known to inhibit fibrinogen/thrombin clotting in mammals. Plasma tended to kill P. dicentrarchi slightly faster when clotting was inhibited by the tetrapeptide, although the total mortality of ciliates was similar. We also found that kaolin, a particulate activator of the intrinsic pathway in mammals, accelerates plasma clotting in turbot. In addition, PMA-stimulated neutrophils, living ciliates and several ciliate components such as cilia, proteases and DNA also displayed procoagulant activity in vitro. Injection of fish with the ciliates generated the massive release of neutrophils to the peritoneal cavity, with formation of large aggregates in those fish with live ciliates in the peritoneum. We observed, by SEM, numerous fibrin-like fibres in the peritoneal exudate, many of which were associated with peritoneal leukocytes and ciliates. Expression of the CD18/CD11b gene, an integrin associated with cell adhesion and the induction of fibrin formation, was upregulated in the peritoneal leukocytes. In conclusion, the findings of the present study show that P. dicentrarchi induces the formation of plasma clots and that the fish coagulation system may play an important role in immobilizing and killing this parasiteThis work was financially supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429 (PARAFISHCONTROL), by the Ministerio de Economía y Competitividad (Spain) under grant agreement AGL2014-57125-R and by grant GPC2014/069 from the Xunta de Galicia (Spain)S