Effects of short-term hyposalinity stress on four commercially important bivalves: A proteomic perspective

Increased heavy rainfall can reduce salinity to values close to 0 in estuaries. Lethal and sublethal physiological and behavioural effects of decreases in salinity below ten have already been found to occur in the commercially important clam species Venerupis corrugata, Ruditapes decussatus and R. philippinarum and the cockle Cerastoderma edule, which generate an income of ∼74 million euros annually in Galicia (NW Spain). However, studies of the molecular response to hyposaline stress in bivalves are scarce. This ‘shotgun’ proteomics study evaluates changes in mantle-edge proteins subjected to short-term hyposaline episodes in two different months (March and May) during the gametogenic cycle. We found evidence that the mantle-edge proteome was more responsive to sampling time than to hyposalinity, strongly suggesting that reproductive stages condition the stress response. However, hyposalinity modulated proteome profiles in V. corrugata and C. edule in both months and R. philippinarum in May, involving proteins implicated in protein folding, redox homeostasis, detoxification, cytoskeleton modulation and the regulation of apoptotic, autophagic and lipid degradation pathways. However, proteins that are essential for an optimal osmotic stress response but which are highly energy demanding, such as chaperones, osmoprotectants and DNA repair factors, were found in small relative abundances. In both months in R. decussatus and in March in R. philippinarum, almost no differences between treatments were detected. Concordant trends in the relative abundance of stress response candidate proteins were also obtained in V. corrugata and C. edule in the different months, but not in Ruditapes spp., strongly suggesting that the osmotic stress response in bivalves is complex and possibly influenced by a combination of controlled (sampling time) and uncontrolled variables. In this paper, we report potential molecular targets for studying the response to osmotic stress, especially in the most osmosensitive native species C. edule and V. corrugata, and suggest factors to consider when searching for biomarkers of hyposaline stress in bivalves.Ministerio de Economía y Competitividad | Ref. CTM2014-51935-RXunta de Galicia | Ref. GRC2013-004Xunta de Galicia | Ref. ED431C 2017/46Xunta de Galicia | Ref. ED431C 2020/05Financiado para publicación en acceso aberto: Universidade de Vigo/CISU

Coagulación intravascular diseminada como forma de presentación de leucemia aguda promielocítica: evaluación y tratamiento

Presentamos el caso de un varón de 29 años ingresado en nuestro centro por hematuriamacroscópica, coagulopatía y leucotrombopenia. Se estableció el diagnóstico inicial de coagulación intravascular diseminada (CID) aguda. El proceso diagnóstico para definir la causa de la CID confirmó la existencia de una Leucemia Promielocítica Aguda (LPA). La coagulopatía asociada a LPA es compleja, con datos biológicos de CID e hiperfibrinolisis primaria. Esta coagulopatía se asocia con hemorragias y se observa en el momento del diagnóstico, agravándose con la quimioterapia antileucémica. Constituye una emergencia médica, con una mortalidad cercana al 10% si no se maneja de forma adecuada. Actualmente, la LPA se trata con agentes no citotóxicos como el Ácido Transretinoico (ATRA) y el Trióxido de Arsénico (ATO), con una disminución de las tasas de sangrado. Por lo que un diagnóstico y tratamiento precoz de la coagulopatía suponen un aumento de la supervivencia en estos pacientes.We present the case of a 29-year-old man admitted to our center for hematuria with coagulopathy, mild leukopenia and moderate thrombopenia. A diagnosis of disseminated intravascular coagulation (DIC) secondary to Acute Promyelocytic Leukemia (APL) was made. Bleeding episodes due to DIC are one of the main causes of mortality at diagnosis in patients with APL. Currently there are lines of treatment with Arsenic Trioxide (ATO) and trans-retinoic acid (ATRA) that have replaced conventional chemotherapy regimens, with a decrease in bleeding rates. Therefore, an early diagnosis and treatment of DIC entails an increase in survival in these patients

The concept of death in children aged from 9 to 11 years: Evidences through inductive and deductive analysis of drawings

The objective of the research was to analyze children’s conceptualization of death through drawings, using a mixed approach, which combines deductive and inductive qualitative analysis. The sample consisted of 99 children aged 9–11 years, who were asked to elaborate a drawing about their idea of death and to explain it to the researchers. Drawings were coded basing on Tamm and Granqvist’s model (deductive analysis) and codes and categories were created and modified (inductive analysis). Three main categories were identified in the analysis and four sub-categories were modified and/or created: causes of death, good death, anxiety-fear and symbolization

Burnout syndrome assessment among Spanish oral surgery consultants:a two populations comparative pilot study

The professional Burnout Syndrome (BOS) or Burnout is considered a professional disease made up of three interrelated dimensions (emotional exhaustion, depersonalization and lack of personal fulfillment). BOS has been documented to most severely affect the healthcare professions, especially dentists. On the other hand, its appearance has been documented at an early age, during dental training. However, there are no studies that analyze its incidence in professionals dedicated to Oral Surgery and Implantology, determining the age of onset and related factors. The modified Maslach questionnaire was carried out anonymously among the professors and students of the Master of Oral Surgery and Implantology at the Complutense University of Madrid. A total of 36 participants were enrolled in this study and the results of the modified Maslach Questionnaire were established into four groups [1st year (n=6), 2nd year (n=6), 3rd year (n=6) postgraduate students and clinical teachers (n=18)]. The following variables were recorded: Age, sex, years of experience, weekly hours of work, dedication on weekends and scope of work. The statistical analysis performed included Pearson's correlation, analysis of variance, Student's t-test, F-Anova, Chi-Square and Gamma correlation. Statistical Significance of the tests was established of p?0.05. 36 questionnaires were analyzed, of which 22.2% (n = 8) presented BOS, and 77.8% (n = 28) a medium risk of suffering it. The mean values and standard deviation ??of emotional exhaustion (7.50 ± 2.43; 9.83 ± 4.12; 15.83 ± 6.21; 30.22 ± 7.86), depersonalization (5.50 ± 1.23; 50 ± 3.27; 11.33 ± 1.75; 17.56 ± 4.13), low personal fulfillment (39.67 ± 3.72; 39.33 ± 2.34; 43.17 ± 3, 55; 37.33 ± 5.51) and professional burnout (54.33 ± 2.66; 61.67 ± 2.88; 70.33 ± 5.43; 85.11 ± 9.05) in the four groups respectively. A significant association was found in the appearance of emotional exhaustion and depersonalization, years of experience, weekly work hours and the work environment. BOS is a disease that can appear from 30 years of age, after 5 years of professional experience and when there is a clinical consultation of 40 hours a week. Oral Surgery and Implantology seems to be a risk activity for the manifestation of depersonalization

Responses to salinity stress in bivalves: Evidence of ontogenetic changes in energetic physiology on Cerastoderma edule

Estuarine bivalves are especially susceptible to salinity fluctuations. Stage-specific sensibilities may influence the structure and spatial distribution of the populations. Here we investigate differences on the energetic strategy of thread drifters (3–4 mm) and sedentary settlers (9–10 mm) of Cerastoderma edule over a wide range of salinities. Several physiological indicators (clearance, respiration and excretion rates, O:N) were measured during acute (2 days) and acclimated responses (7 days of exposure) for both size classes. Our results revealed a common lethal limit for both developmental stages (Salinity 15) but a larger physiological plasticity of thread drifters than sedentary settlers. Acclimation processes in drifters were initiated after 2 days of exposure and they achieved complete acclimation by day 7. Sedentary settlers delay acclimation and at day 7 feeding activity had not resumed and energetic losses through respiration and excretion were higher at the lowest salinity treatment. Different responses facing salinity stress might be related to differences in habitat of each stage. For sedentary settlers which occupy relatively stable niches, energy optimisation include delaying the initiation of the energetically expensive acclimation processes while drifters which occupy less stable environments require a more flexible process which allow them to optimize energy acquisition as fast as possible.Ministerio de Economía y Competitividad | Ref. CTM2014–51935-RXunta de Galicia | Ref. POS-B/2016/032Xunta de Galicia | Ref. GRC2013–00

GNIP1 E3 ubiquitin ligase is a novel player in regulating glycogen metabolism in skeletal muscle.

Background: Glycogenin-interacting protein 1 (GNIP1) is a tripartite motif (TRIM) protein with E3 ubiquitin ligase activity that interacts with glycogenin. These data suggest that GNIP1 could play a major role in the control of glycogen metabolism. However, direct evidence based on functional analysis remains to be obtained. Objectives: The aim of this study was 1) to define the expression pattern of glycogenin-interacting protein/ Tripartite motif containing protein 7 (GNIP/TRIM7) isoforms in humans, 2) to test their ubiquitin E3 ligase activity, and 3) to analyze the functional effects of GNIP1 on muscle glucose/glycogen metabolism both in human cultured cells and in vivo in mice. Results: We show that GNIP1 was the most abundant GNIP/TRIM7 isoform in human skeletal muscle, whereas in cardiac muscle only TRIM7 was expressed. GNIP1 and TRIM7 had autoubiquitination activity in vitro and were localized in the Golgi apparatus and cytosol respectively in LHCN-M2 myoblasts. GNIP1 overexpression increased glucose uptake in LHCN-M2 myotubes. Overexpression of GNIP1 in mouse muscle in vivo increased glycogen content, glycogen synthase (GS) activity and phospho-GSK-3α/β (Ser21/9) and phospho-Akt (Ser473) content, whereas decreased GS phosphorylation in Ser640. These modifications led to decreased blood glucose levels, lactate levels and body weight, without changing whole-body insulin or glucose tolerance in mouse. Conclusion: GNIP1 is an ubiquitin ligase with a markedly glycogenic effect in skeletal muscle

Vertidos tóxicos al río Guadiamar: propuestas técnicas para su corrección

Inmediatamente de producirse el vertido tóxico al río Guadiamar, el Grupo T.A.R. se lanzó sin pensarlo dos veces a la búsqueda de soluciones técnicas a un panorama desolador y de efectos desconocidos, todos ellos amenazantes. El ácido “se comía el suelo inundado” por la riada, el agua retenida en Entremuros a pH 3, y con un enorme contenido de metales pesados, ocupaba una extensión de kilómetros. Nos hundimos en el agua hasta el cuello, y cuando nos cubría cogimos la barca, metimos el río a pedazos en nuestro laboratorio, para trabajar todas las hipótesis, ensayar todas las posibilidades. Peleando con la realidad le sacamos datos al Guadiamar, diseñamos actuaciones, poniéndole ingeniería a cuantas hipótesis nos planteaba la situación. En primera fila observamos las mejores actuaciones que nadie diseñó. El propio río, activando sus defensas naturales, mejoró la calidad del agua retenida en el dique de Entremuros subiendo el pH y precipitando los metales pesados. Los mecanismos de entrada de los metales pesados en la cadena trófica parecían ser lentos, dando tiempo a que la retirada de los lodos tóxicos llevada a cabo por la Administración fuera eficaz y diera tiempo a realizar tanto esfuerzo. Aunque el Guadiamar ha trabajado muy duro en su propia recuperación, con su ayuda hemos elaborado una gran cantidad de propuestas técnicas; unas para actuaciones de emergencia, otras a corto, medio y largo plazo. También hemos dado forma a un Plan frente a las previsibles avenidas de este primer otoño después del vertido. Nuestro objetivo ha sido poner a disposición soluciones preparadas para todo tipo de problemas, en primera o en segunda instancia. Prevenir no solo una o dos contingencias, se ha tratado de estar preparado para la mayor cantidad de eventualidades posibles. Por ello algunas serán utilizables, otras estarán en reserva, y muchas irían al cajón de los papeles. Pero ahí están por si acaso. Este libro recoge los trabajos de campo, los ensayos de laboratorio y la ingeniería desarrollada en los primeros cuatro meses. Durante el siguiente preparamos la edición del mismo, mientras, en paralelo, continuábamos en el trabajo experimental y el diseño. Cuando se cumpla el quinto mes, el 25 de Septiembre de 1998, lo presentaremos, ciento cincuenta días después... Con la financiación de la Diputación de Sevilla hemos preparado la primera edición en formato CD Rom e Internet, con muy poco coste para acceder a su contenido. En poco tiempo saldrá la edición en papel, con la misma financiación que la primera. Nos gustaría que este documento fuera entendido como lo que es, en nuestra opinión, una llamada urgente al debate de las ideas. Tratamos de ofrecer la información necesaria y el foro donde recoger las propuestas que seguramente muchos pueden aportar sin saber como transmitir sus experiencias. El Grupo de Tratamiento de Aguas Residuales (T.A.R.) abre con este libro la MESA DE DISCUSIÓN, para buscar un poco de luz, avanzar en las soluciones técnicas a la inmensa tarea de recuperar el río Guadiamar. El libro presenta lagunas, unas por la enorme prisa, otra por falta de datos, muchas por nuestra escasez de conocimientos. Dicen en España que “lo mejor es enemigo de lo bueno”...,y nos gustaría recoger ideas hoy mejor que mañana, que podría ser tarde. Nos comprometemos a seguir trabajando en soluciones técnicas, innovaciones tecnológicas e investigación aplicada a la recuperación del Guadiamar, a conocer lo ocurrido y su remedio. Nos comprometemos a publicar de la misma forma los resultados obtenidos, de manera que la discusión y el debate sigan siempre abiertos. El grupo T.A.R. podría ser un punto de intercambio de conocimientos universal, abierto, respetuoso y tolerante, universitario en definitiva, y por tanto útil en el cumplimiento de sus obligaciones. La primera necesidad de responder urgentemente, está dando paso a unas actuaciones programadas, a medida de los efectos de las correcciones introducidas. Deben instaurarse políticas de prevención y nuevas actuaciones para recuperar el Guadiamar, mejorar urgentemente las condiciones del entorno. Aprender de las soluciones adoptadas y generar mejores prácticas, puede ser una buena conclusión del trabajo realizado por tanta gente. Lo que empezó siendo una carrera de velocidad se nos convierte en un maratón, ya no hay que correr explosivamente, hay que mantener un ritmo en la carrera; hay que persistir en el esfuerzo todos los días durante mucho tiempo. Este nuevo desafío sigue siendo duro y difícil. Podéis contar con el Grupo T.A.R. para recorrer el duro camino de la Recuperación

Vaccine breakthrough infections with SARS-CoV-2 Alpha mirror mutations in Delta Plus, Iota, and Omicron

Replication of SARS-CoV-2 in the human population is defined by distributions of mutants that are present at different frequencies within the infected host and can be detected by ultra-deep sequencing techniques. In this study, we examined the SARS-CoV-2 mutant spectra of amplicons from the spike-coding (S-coding) region of 5 nasopharyngeal isolates derived from patients with vaccine breakthrough. Interestingly, all patients became infected with the Alpha variant, but amino acid substitutions that correspond to the Delta Plus, Iota, and Omicron variants were present in the mutant spectra of the resident virus. Deep sequencing analysis of SARS-CoV-2 from patients with vaccine breakthrough revealed a rich reservoir of mutant types and may also identify tolerated substitutions that can be represented in epidemiologically dominant variants.This work was supported by the Instituto de Salud Carlos III, Spanish Ministry of Science and Innovation (COVID-19 Research Call COV20/00181) and co-financed by the European Development Regional Fund “A way to achieve Europe.” The work was also supported by grants CSIC-COV19-014 from the CSIC, project 525/C/2021 from the Fundació La Marató de TV3; PID2020-113888RB-I00 from the Ministerio de Ciencia e Innovación; BFU2017-91384-EXP from the Ministerio de Ciencia, Innovación y Universidades (MCIU);PI18/00210 and PI21/00139 from the Instituto de Salud Carlos III; and S2018/BAA-4370 (PLATESA2) from the Comunidad de Madrid/ FEDER. This research work was also funded by the European Commission – NextGenerationEU (regulation EU 2020/2094), through the CSIC’s Global Health Platform (PTI Salud Global). CP and PM are supported by the Miguel Servet programme of the Instituto de Salud Carlos III (CPII19/00001 and CP16/00116, respectively), cofinanced by the European Regional Development Fund (ERDF). CIBERehd is funded by the Instituto de Salud Carlos III. Institutional grants from the Fundación Ramón Areces and Banco Santander to the CBMSO are also acknowledged. The team at CBMSO belongs to the Global Virus Network (GVN). BMG is supported by predoctoral contract PFIS FI19/00119 from the Instituto de Salud Carlos III (Ministerio de Sanidad y Consumo), cofinanced by the Fondo Social Europeo (FSE). CGC is supported by predoctoral contract PRE2018- 083422 from the MCIU. BS was supported by a predoctoral research fellowship (Doctorados Industriales, DI-17-09134) from the Spanish Ministry of Economy and Competitiveness (MINECO).Peer reviewe

Amino acid substitutions associated with treatment failure of hepatitis C virus infection

Trabajo presentado en el XVI Congreso Nacional de Virología, celebrado en Málaga (España) del 06 al 09 de septiembre de 2022.Despite the high sustained virological response rates achieved with current directly-acting antiviral agents (DAAs) against hepatitis C virus (HCV), around 2% to 5% of patients do not achieve such a response. Identification of amino acid substitutions associated with treatment failure requires analytical designs, such as subtype-specific ultra-deep sequencing (UDS) methods for HCV characterization and patient management. By deep sequencing analysis of 220 subtyped HCV samples from infected patients who failed therapy, collected from 39 Spanish hospitals, we determined amino acid sequences of the DAA-target proteins NS3, NS5A and NS5B, by UDS of HCV patient samples, in search of resistanceassociated substitutions (RAS). Using this procedure, we have identified six highly represented amino acid substitutions (HRSs) in NS5A and NS5B of HCV, which are not bona fide RAS. They were present frequently in basal and post-treatment virus of patients who failed therapy to different DAA-based therapies. Contrary to several RAS, HRSs belong to the acceptable subset of substitutions according to the PAM250 replacement matrix. Coherently, their mutant frequency, measured by the number of deep sequencing reads within the HCV quasispecies that encode the relevant substitutions, ranged between 90% and 100% in most cases. Also, they have limited predicted disruptive effects on the threedimensional structures of the proteins harboring them. The information on HRSs that will be gathered during sequencing should be relevant not only to help predict treatment outcomes and disease progression but also to further understand HCV population dynamics, which appears much more complex than thought prior to the introduction of deep sequencing.The work at CBMSO was supported by grants SAF2014-52400-R from MINECO, SAF2017-87846-R and BFU2017-91384-EXP MICIU, PI18/00210 from ISCIII, S2013/ABI-2906 (PLATESA) and S2018/BAA-4370 (PLATESA2) from Comunidad de Madrid/FEDER. C.P. is supported by the Miguel Servet program of the ISCIII (CP14/00121 and CPII19/00001), cofinanced by the European Regional Development Fund (ERDF). CIBERehd is funded by ISCIII. Institutional grants from the Fundación Ramón Areces and Banco Santander to the CBMSO are also acknowledged. The team at CBMSO belongs to the Global Virus Network (GVN). The work in Barcelona was supported by ISCIII, cofinanced by ERDF grant number PI19/00301 and by the Centro para el Desarrollo Tecnológico Industrial (CDTI) from the MICIU, grant number IDI20151125. Work at CAB was supported by MINECO grant BIO2016-79618R and PID2019-104903RB-I00 (funded by the EU under the FEDER program) and by the Spanish State research agency (AEI) through project number MDM-2017-0737 Unidad de Excelencia “María de Maeztu”-Centro de Astrobiología (CSIC-INTA). Work at IBMB was supported by MICIN grant BIO2017-83906-P (funded by the EU under the FEDER program). C.G.-C. is supported by predoctoral contract PRE2018-083422 from MICIU. B.M.-G. is supported by predoctoral contract PFIS FI19/00119 from Instituto de Salud Carlos III (Ministerio de Sanidad y Consumo), cofinanced by Fondo Social Europeo (FSE).Peer reviewe