Search

AIM The aim was to investigate the influence of distal resection margin and extent of mesorectal excision on long-term oncological outcomes. METHOD Consecutive patients with upper and middle third rectal cancer from June 2006 to February 2016 were reviewed. Patients were divided into four groups depending on the distal margin considered as a surrogate marker of the extension of mesorectal excision (Q1 ≤10 mm, Q2 11-20 mm, Q3 21-30 mm, Q4 ≥31 mm). Local-recurrence-free survival (LRFS), disease-free survival (DFS) and overall survival (OS) were estimated. Cox regression models were used to investigate the influence of surgical and clinicopathological variables on prognosis by adjusting for confounding factors. RESULTS Two hundred and eleven patients with mid (125) and upper (86) rectal cancer underwent wide mesorectal excision. The median follow-up was 48.64 months (interquartile range 28-63). 17.5% patients developed recurrence. The 5-year LRFS, DFS and OS for all patients were 93.20%, 83.89% and 80.1%, respectively, with no statistically significant differences between groups (LRFS, P = 0.601; DFS, P = 0.487; OS, P = 0.468). In the multivariable analysis the recurrences and survival were associated with the quality of the mesorectum (LRFS, hazard ratio 10.629, 95% CI 2.324-48.610, P = 0.002; DFS, hazard ratio 2.789, 95% CI 1.314-5.922, P = 0.008). CONCLUSION A wide anatomical resection with partial mesorectal excision and shorter distal resection margin does not jeopardize the oncological outcomes.

13 pags., 7 figs., 1 tab., Owing to their outstanding catalytic properties, enzymes represent powerful tools for carrying out a wide range of (bio)chemical transformations with high proficiency. In this context, enzymes with high biocatalytic promiscuity are somewhat neglected. Here, we demonstrate that a meticulous modification of a synthetic shell that surrounds an immobilized enzyme possessing broad substrate specificity allows the resulting nanobiocatalyst to be endowed with enantioselective properties while maintaining a high level of substrate promiscuity. Our results show that control of the enzyme nano-environment enables tuning of both substrate specificity and enantioselectivity. Further, we demonstrate that our strategy of enzyme supramolecular engineering allows the enzyme to be endowed with markedly enhanced stability in an organic solvent (i.e., acetonitrile). The versatility of the method was assessed with two additional substrate-promiscuous and structurally different enzymes, for which improvements in enantioselectivity and stability were confirmed. We expect this method to promote the use of supramolecularly engineered promiscuous enzymes in industrially relevant biocatalytic processes., We thank the staff of the Synchrotron Radiation Source at Alba (Barcelona, Spain) for assistance at the BL13-XALOC beamline. The authors would like to acknowledge M. Chami and the Bio-EM lab of the grant agreement no. 634486), grants PCIN-2017-078 (within the Marine Biotechnology ERA-NET), BIO2016-76601-C3-3- R, and BIO2017-85522-R from the Ministerio de Economía y Competitividad, Ministerio de Ciencia, Innovacion y Uni- ́ versidades (MCIU), Agencia Estatal de Investigacion (AEI), ́ Fondo Europeo de Desarrollo Regional (FEDER), and European Union (EU). R.B. acknowledges the support of the UK Biotechnology and Biological Sciences Research Council (BBSRC; grant No. BB/M029085/1) and the Centre of Environmental Biotechnology Project and the Supercomputing Wales project, which are partly funded by the European Regional Development Fund (ERDF) via the Welsh Government

Autophagy is a key process in the maintenance of cellular homeostasis. The age-dependent decline in retinal autophagy has been associated with photoreceptor degeneration. Retinal dysfunction can also result from damage to the retinal pigment epithelium (RPE), as the RPE–retina constitutes an important metabolic ecosystem that must be finely tuned to preserve visual function. While studies of mice lacking essential autophagy genes have revealed a predisposition to retinal degeneration, the consequences of a moderate reduction in autophagy, similar to that which occurs during physiological aging, remain unclear. Here, we describe a retinal phenotype consistent with accelerated aging in mice carrying a haploinsufficiency for Ambra1, a pro-autophagic gene. These mice show protein aggregation in the retina and RPE, metabolic underperformance, and premature vision loss. Moreover, Ambra1+/gt mice are more prone to retinal degeneration after RPE stress. These findings indicate that autophagy provides crucial support to RPE-retinal metabolism and protects the retina against stress and physiological aging.

641 Background: Immune checkpoint inhibitors have become a cornerstone in the management of mRCC. However, to identify the most suitable patients for this treatment is an unmet medical need. We aimed to explore the utility of DL integrating clinical and molecular data to predict response to immunotherapy. Methods: We conducted a retrospective analysis using publicly available data from patients treated with nivolumab in the clinical trials Checkmate 009, 010 and 025. The primary objective was to assess the performance of different DL models (autoencoder and convolutional neural network (CNN)) predicting the PFS of these patients. With that scope, we followed several research lines including the creation of combined datasets with clinic and RNA sequencing and comparing the results of the DL models against the performance of traditional machine learning (ML) models. Finally, we came up with an interpretability analysis of those black-box models using LIME and SHAP values. Results: Clinical and transcriptomic data were available from 181 nivolumab-treated patients. Outcomes achieved confirmed that we can model the response for NIVOLUMAB using RNA sequencing data. (Table) However, DL models have not demonstrated to be significantly better than traditional ML methods when predicting response (p= 0.068). Deep autoencoder provided 68.9% accuracy, but the most accurate model was logistic regression classifier which achieved 86.4% of hit rate. Interpretability results revealed that most relevant genes for the decision making were related with the immune response and the regulation of kinases. Regarding interpretability, best results were achieved integrating both transcriptomic and clinical data (7 out of 10 DL and ML tested models achieved higher hit rates with combined data set. Logistic regression classifier improved its accuracy in 11%). Conclusions: The integration of clinical and molecular data could lead to more accurate predictions of outcome than any dataset by its own. However, further research is intended in the field of the DL analysis, as data codification and data structure could bias the results. The ongoing study ART (Artificial Intelligence in Renal Tumors) will address this issue prospectively. [Table: see text]

Context Derivation of embryonic stem cells (ESC) from single blastomeres is an interesting alternative to the use of whole blastocysts, but derivation rates are lower and the requirements for successful ESC obtention are still poorly defined. Aims To investigate the effects of embryo cryopreservation and of signalling modulators present during embryo culture and/or ESC establishment on ESC derivation efficiency from single 8-cell mouse blastomeres. Method Fresh and cryopreserved 2-cell embryos were cultured and biopsied at the 8-cell stage. Single blastomeres were cultured in the presence of 2i or R2i cocktails, with or without adrenocorticotropic hormone (ACTH). We analysed ESC derivation efficiencies and characterised pluripotency genes expression and karyotype integrity of the resulting lines. We also evaluated the impact of embryo preculture with R2i on epiblast cell numbers and derivation rates. Key results The ESC generation was not compromised by embryo cryopreservation and ACTH was dispensable under most of the conditions tested. While 2i and R2i were similarly effective for ESC derivation, R2i provided higher karyotype integrity. Embryo preculture with R2i yielded increased numbers of epiblast cells but did not lead to increased ESC generation. Conclusions Our findings help to define a simplified and efficient procedure for the establishment of mouse ESC from single 8-cell blastomeres. Implications This study will contribute to improving the potential of this experimental procedure, providing a tool to investigate the developmental potential of blastomeres isolated from different embryonic stages and to reduce the number of embryos needed for ESC derivation.

Fundació Carreras This work was supported by Spanish Ministry of Science and Innovation (PID2020-117645RB-I00/AEI/10.13039/501100011033 and RYC2020-029400-I/AEI/10.13039/501100011033), the FERO Foundation (BFero2020.03), and the Government of Andorra (ATC025-AND-2020). We thank the CERCA Program (Generalitat de Catalunya), the Josep Carreras Leukemia Research Institute and the Catalan Institute of Oncology for their institutional support. LB is supported by the Spanish Ministry of Science and Innovation (MCI) (PID2020-117645RB-I00/AEI/10.13039/501100011033 and RYC2020-029400-I/AEI/10.13039/501100011033), and the FERO Foundation (BFero2020.03). CF is supported by a PhD grant from the Government of Andorra (ATC025-AND-2020). Juvenile myelomonocytic leukemia (JMML) is a rare myelodysplastic/myeloproliferative neoplasm of early childhood. Most of JMML patients experience an aggressive clinical course of the disease and require hematopoietic stem cell transplantation, which is currently the only curative treatment. JMML is characterized by RAS signaling hyperactivation, which is mainly driven by mutations in one of five genes of the RAS pathway, including PTPN11, KRAS, NRAS, NF1, and CBL. These driving mutations define different disease subtypes with specific clinico-biological features. Secondary mutations affecting other genes inside and outside the RAS pathway contribute to JMML pathogenesis and are associated with a poorer prognosis. In addition to these genetic alterations, JMML commonly presents aberrant epigenetic profiles that strongly correlate with the clinical outcome of the patients. This observation led to the recent publication of an international JMML stratification consensus, which defines three JMML clinical groups based on DNA methylation status. Although the characterization of the genomic and epigenomic landscapes in JMML has significantly contributed to better understand the molecular mechanisms driving the disease, our knowledge on JMML origin, cell identity, and intratumor and interpatient heterogeneity is still scarce. The application of new single-cell sequencing technologies will be critical to address these questions in the future.

22 p.-9 fig., Macroautophagy/autophagy is a key process in the maintenance of cellular homeostasis. The age-dependent decline in retinal autophagy has been associated with photoreceptor degeneration. Retinal dysfunction can also result from damage to the retinal pigment epithelium (RPE), as the RPE-retina constitutes an important metabolic ecosystem that must be finely tuned to preserve visual function. While studies of mice lacking essential autophagy genes have revealed a predisposition to retinal degeneration, the consequences of a moderate reduction in autophagy, similar to that which occurs during physiological aging, remain unclear. Here, we described a retinal phenotype consistent with accelerated aging in mice carrying a haploinsufficiency for Ambra1, a pro-autophagic gene. These mice showed protein aggregation in the retina and RPE, metabolic underperformance, and premature vision loss. Moreover, Ambra1+/gt mice were more prone to retinal degeneration after RPE stress. These findings indicate that autophagy provides crucial support to RPE-retinal metabolism and protects the retina against stress and physiological aging., This work was supported by the H2020 Excellent Science [No. 765912]; Instituto de Salud Carlos III,CiberNed [CB06/05/0041]; Spanish Ministry of Science and Innovation [RTI2018-098990-J-I00]; Spanish Ministry of Science and Innovation [PGC2018-098557-B-I00].

Parkinson’s disease (PD) is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Since there are only symptomatic treatments available, new cellular and molecular targets involved in the onset and progression of this disease are needed to develop effective treatments. CCAAT/Enhancer Binding Protein β (C/EBPβ) transcription factor levels are altered in patients with a variety of neurodegenerative diseases, suggesting that it may be a good therapeutic target for the treatment of PD. A list of genes involved in PD that can be regulated by C/EBPβ was generated by the combination of genetic and in silico data, the mitochondrial transcription factor A (TFAM) being among them. In this paper, we observed that C/EBPβ overexpression increased TFAM promoter activity. However, downregulation of C/EBPβ in different PD/neuroinflammation cellular models produced an increase in TFAM levels, together with other mitochondrial markers. This led us to propose an accumulation of non-functional mitochondria possibly due to the alteration of their autophagic degradation in the absence of C/EBPβ. Then, we concluded that C/EBPβ is not only involved in harmful processes occurring in PD, such as inflammation, but is also implicated in mitochondrial function and autophagy in PD-like conditions.

Introduction Diverting stomata are recommended in patients with low anterior resection and risk factors in order to reduce the severity of anastomotic leaks. Usually, a radiology study is performed prior to the closure of the stoma to detect subclinical leaks. The aim of the present study is to assess the clinical utility of the radiology study. Methods A prospective cohort study of patients undergoing anterior rectal resection for rectal cancer and those who underwent stoma closure without contrast enema. This study was carried out after a retrospective review of radiology study results prior to the closure of the stoma in patients operated from 2007 to 2011. Results Eighty-six patients met the study criteria. Thirteen patients (15.1%) presented pelvic sepsis. Contrast enema before stoma closure was pathological in 8 patients (9.3%). Five out of the 13 patients with pelvic sepsis had a pathological radiological study, compared to only 3 out of the 73 patients without intra-abdominal complications after rectal resection (38.5% vs 4.1%; P=.001). Based on these results, we conducted a prospective study omitting the contrast enema in patients with no postoperative complications. Thirty-eight patients had their stoma closed without a prior radiology study. None of the patients presented pelvic sepsis. Conclusions Radiology studies of the colorectal anastomosis before reconstruction can safely be omitted in patients without pelvic sepsis after the previous rectal resection.

Resumen Introduccion Es recomendable practicar un estoma derivativo en pacientes con reseccion anterior baja y factores de riesgo, para reducir la gravedad de la fuga anastomotica. Habitualmente se realiza un estudio radiologico previo al cierre del estoma para detectar fugas subclinicas. El objetivo del presente estudio es evaluar la utilidad clinica del estudio radiologico. Metodos Estudio prospectivo de una cohorte de pacientes sometidos a reseccion anterior de recto por cancer rectal, y a los que se les realiza cierre del estoma sin enema de contraste. Este estudio se lleva a cabo despues de realizar una revision retrospectiva sobre los resultados del estudio radiologico previo al cierre del estoma en pacientes intervenidos entre 2007 y 2011. Resultados Ochenta y seis pacientes cumplieron los criterios del estudio. Trece pacientes (15,1%) presentaron sepsis pelvica. El enema con contraste antes del cierre del estoma fue patologico en 8 pacientes (9,3%). Cinco de los 13 pacientes con sepsis pelvica mostraron un estudio radiologico patologico, en comparacion con solo 3 de 73 pacientes sin complicaciones intraabdominales despues de la reseccion del recto (38,5% vs 4,1%; p = 0,001). Basandonos en estos resultados, realizamos un estudio prospectivo omitiendo el enema con contraste en pacientes con curso postoperatorio no complicado. A 38 pacientes se les cerro el estoma sin estudio radiologico previo. Ningun paciente presento sepsis pelvica. Conclusiones El estudio radiologico de la anastomosis colorrectal antes de la reconstruccion del transito puede omitirse con seguridad en los pacientes sin sepsis pelvica ni ileo paralitico tras la reseccion anterior de recto.

NOTCH1 is a well-established lineage specifier for T cells and among the most frequently mutated genes throughout all subclasses of T cell acute lymphoblastic leukemia (T-ALL). How oncogenic NOTCH1 signaling launches a leukemia-prone chromatin landscape during T-ALL initiation is unknown. Here we demonstrate an essential role for the high-mobility-group transcription factor Tcf1 in orchestrating chromatin accessibility and topology, allowing aberrant Notch1 signaling to convey its oncogenic function. Although essential, Tcf1 is not sufficient to initiate leukemia. The formation of a leukemia-prone epigenetic landscape at the distal Notch1-regulated Myc enhancer, which is fundamental to this disease, is Tcf1-dependent and occurs within the earliest progenitor stage even before cells adopt a T lymphocyte or leukemic fate. Moreover, we discovered a unique evolutionarily conserved Tcf1-regulated enhancer element in the distal Myc-enhancer, which is important for the transition of preleukemic cells to full-blown disease.

Mestrado em Auditoria Submitted by Filomena Borba (fmborba@iscal.ipl.pt) on 2021-05-07T16:42:54Z No. of bitstreams: 1 Dissertaá∆o Mestrado_Vers∆o_Definitiva_Sandra Diogo_20180304_05_03_2021.pdf: 2846754 bytes, checksum: b467e54722eee60905c717dcb97b9099 (MD5) Made available in DSpace on 2021-05-07T16:42:54Z (GMT). No. of bitstreams: 1 Dissertaá∆o Mestrado_Vers∆o_Definitiva_Sandra Diogo_20180304_05_03_2021.pdf: 2846754 bytes, checksum: b467e54722eee60905c717dcb97b9099 (MD5) Previous issue date: 2021-03 info:eu-repo/semantics/publishedVersion

There is a consensus among researchers about the link between low meaning in life and anxiety and depressive symptoms. One unanswered question is whether meaning-making is a mediator of the change in anxiety and depression symptoms in participants with adjustment disorders during cognitive behavioural therapy (CBT) treatment. The aims of this study were (a) to analyse whether there was meaning-making during the application of the CBT, (b) to analyse whether meaning-making was a mediator of anxiety psychopathology and (c) to analyse whether meaning-making was a mediator of depressive symptoms. The sample was composed of 115 patients who satisfied the full Diagnostic and Statistical Manual of Mental Disorders (DSM)-5 criteria for adjustment disorder as their primary diagnosis and completed CBT treatment in a primary care mental health service: 74.78% women, n = 86, and 25.22% men, n = 29, with a mean age of 41.89 (standard deviation [SD] = 10.39) years. The diagnosis was established using the Structured Clinical Interview for DSM-5 (SCID-5), and participants filled out the Beck Anxiety Inventory, the Beck Depression Inventory and Purpose in Life questionnaires. The therapists were clinical psychologists with experience in clinical assessment. A repeated-measures analysis of variance (ANOVA) and two mediation analyses using the bootstrap method were performed. The results indicated that (a) There was meaning-making during the CBT because the treated sample showed a statistically significant improvement in meaning in life, and (b) meaning-making during the CBT was a partial mediator between anxiety symptoms and depressive symptoms before and after the treatment. The present study suggests that meaning in life could be an important variable in the psychopathology of adjustment disorders.

Individuals who suffer from depressive symptoms experience a substantial impact on psychosocial functioning, physical health, mortality, and quality of life. In the search for therapeutic strategies, exercise has been found to play a relevant part in its treatment. However, the promotion of exercise entails adherence difficulties that arose out of the tendency towards sedentarism led by symptomatology. Personalised exercise plans on top of usual care have the potential to enhance behavioural changes and mental health. The present study aims at evaluating the changes in functioning deriving from a blended intervention merging a psychological intervention with a personalised exercise programme based on medical assessment. We will conduct a three-arm randomised controlled trial in which 172 participants suffering from mild–moderate depressive symptoms will be allocated to Intervention A (personalised exercise group programme + app with motivational messages), B (personalised exercise group programme + app with no motivational messages) or control group (app with no motivational messages). Data regarding global functioning, well-being, symptoms, physical activity, and exercise capacity will be collected at baseline, 4, 12, and 36 weeks. The results of this trial will provide information about whether this physical activity support programme may be efficient for improving mental and physical health outcomes. Trial registration: ClinicalTrials.gov NCT04857944 (accessed on 15 April 2021). Registered April 2021.

N,N-dimethyltryptamine (DMT) is a component of the ayahuasca brew traditionally used for ritual and therapeutic purposes across several South American countries. Here, we have examined, in vitro and vivo, the potential neurogenic effect of DMT. Our results demonstrate that DMT administration activates the main adult neurogenic niche, the subgranular zone of the dentate gyrus of the hippocampus, promoting newly generated neurons in the granular zone. Moreover, these mice performed better, compared to control non-treated animals, in memory tests, which suggest a functional relevance for the DMT-induced new production of neurons in the hippocampus. Interestingly, the neurogenic effect of DMT appears to involve signaling via sigma-1 receptor (S1R) activation since S1R antagonist blocked the neurogenic effect. Taken together, our results demonstrate that DMT treatment activates the subgranular neurogenic niche regulating the proliferation of neural stem cells, the migration of neuroblasts, and promoting the generation of new neurons in the hippocampus, therefore enhancing adult neurogenesis and improving spatial learning and memory tasks.

PURPOSE: This study aimed to determine the role of Wnt pathway in mouse embryonic stem cell (mESC) derivation from single blastomeres isolated from eight-cell embryos and in the pluripotency features of the mESC established. METHODS: Wnt activator CHIR99021, Wnt inhibitor IWR-1-endo, and MEK inhibitor PD0325901 were used alone or in combination during ESC derivation and maintenance from single blastomeres biopsied from eight-cell embryos. Alkaline phosphatase activity, FGF5 levels, expression of key pluripotency genes, and chimera formation were assessed to determine the pluripotency state of the mESC lines. RESULTS: Derivation efficiencies were highest when combining pairs of inhibitors (15–24.7%) than when using single inhibitors or none (1.4–10.1%). Full naïve pluripotency was only achieved in CHIR- and 2i-treated mESC lines, whereas IWR and PD treatments or the absence of treatment resulted in co-existence of naïve-like and primed-like pluripotency features. IWR + CHIR- and IWR + PD-treated mESC displayed features of primed pluripotency, but IWR + CHIR-treated lines were able to generate germline-competent chimeric mice, resembling the predicted properties of formative pluripotency. CONCLUSION: Wnt and MAPK pathways have a key role in the successful derivation and pluripotency features of mESC from single precompaction blastomeres. Modulation of these pathways results in mESC lines with various degrees of naïve-like and primed-like pluripotency features.

espanolObjetivos Determinar la incidencia de hernia incisional (HI) en la incision de asistencia (IA) de la pieza en cirugia por neoplasia de colon y recto. Analisis de la relacion de la localizacion de la incision y uso de una malla en la prevencion de la HI en pacientes de alto riesgo. Metodos Revision retrospectiva de la base de datos de cirugia de colon entre enero de 2015 y diciembre de 2016. Se establecieron 2 grupos: incision transversa (IT) e incision media (IM), a su vez este con 2 subgrupos (malla [IMM] y sutura [IMS]). Se categorizaron los pacientes mediante el sistema HERNIAscore. Las hernias se diagnosticaron clinicamente y por TAC. Resultados Se intervino a 210 pacientes, de los que fueron incluidos 182. Tras un seguimiento de 13,0 meses, se detectaron un total de 39 HI (21,9%), de las que 23 (13,4%) fueron en las IA. Estas fueron mucho menos frecuentes en el grupo de IT (3,4%) y en el de IMM (5,9%) que en el de IMS (29,5%; p = 0,007). La probabilidad de aparicion en el grupo IMS de una HI presento una OR = 11,7 (IC 95%: 3,3-42,0) frente a las IT y de 4,3 (IC 95%: 1,1-16,3) frente al grupo IMM. Conclusiones La localizacion de la incision es relevante para disminuir las HI. La IT deberia ser utilizada preferentemente. En los casos en que se utilice una IM, el uso de una malla profilactica en pacientes de alto riesgo puede considerarse, ya que es seguro y con baja morbilidad. EnglishObjectives To determine the incidence of incisional hernia (IH) in the extraction incision (EI) in colorectal resection for cancer. To analyze whether the location of the incision has any relationship with the incidence of hernias and whether mesh could be useful for prevention in high-risk patients. Methods Retrospective review of the colon and rectal surgery database from January 2015 to December 2016. Data were classified into 2 groups, transverse (TI) and midline incision (MI), and the latter was divided into 2 subgroups (mesh [MIM] and suture [MIS]). Patients were classified using the HERNIAscore. Hernias were diagnosed by clinical and/or CT examination. Results A total of 182 out of 210 surgical patients were included. After a median follow-up of 13.0 months, 39 IH (21.9%) were detected, 23 of which (13.4%) were in the EI; their frequency was lower in the TI group (3.4%) and in the MIM group (5.9%) than in the MIS group (29.5%; p = 0.007). The probability of developing IH in the MIS group showed an OR = 11.7 (95%CI: 3.3-42.0) compared to the TI group and 4.3 (IC 95%: 1.1-16.3) versus the MIM group. Conclusions The location of the incision is relevant to avoid incisional hernias. Transverse incisions should be used as the first option. When a midline incision is needed, a prophylactic mesh could be considered in high risk patients because it is safe and associated with low morbidity.

Global attempts to renew scientific education aim to stop the decline of young people's interest in science and technology, and to promote the development of citizens' scientific literacy for sustainable development. Among other changes, these aims require the adaptation of K12 Biological Science Teacher’s training to meet the new objectives.Scientific literacy involves knowing science and how knowledge is developed and validated, recognizing the interactions between science, technology and society, that is, the nature of science (NoS; a set of meta-scientific contents that encompass historical, epistemological and sociological aspects of science with great value for scientific education). It also involves grasping of cognitive skills underlying critical thinking (CT; a set of cognitive abilities, including self-regulation and metacognitive processes) and creative problem solving. Therefore, scientific literacy contributes to making informed decisions, facilitating the participation of citizens in situations and dilemmas of scientific tenor.In addition, CT is closely related to the performance of educators in their professional work. Particularly, in the teaching of science, CT skills favour and enhance the learning of concepts and theories linked not only to science but also to the NoS.Considering the current conditions of middle-higher K12 Biological Science Teachers’ formation and classrooms´ limitations in our country and the region, we propose a pilot project aiming to promote the transformation of initial teachers´ training, seeking to improve the development of CT skills and to deepen NoS comprehension. It will involve the immersion of K12 Biological Science Teacher students in Biological Science Living Physical and "mirror" Virtual Reality Laboratories. These laboratories will be equipped with "do it yourself" (DIY), "do it with others" (DIWO) and "bring your own device" (BYOD) technologies for the implementation of research-type activities framed in the philosophy of the "fabrication laboratories". The virtual platform will also comprise a library with didactic resources under permissive licenses to ensure a broader impact.Within these environments, K12 Biological Science Teacher students will engage in the creation of didactic units involving problem solving and knowledge building in parallel to deeper understanding of scientific processes. We also hope to promote the creativity and innovation of the participants, and the appropriation of DIY/DIWO/BYOD and virtual reality technologies as educational resources in the classroom and everyday life.Thanks to the virtual environment, this approach would also allow to reach both National and International K12 Biological Science Teacher students and graduate Biological Science Teachers. Considering the universal access to the Internet and free access to educational platforms in several countries, we also hope to impact on the non-formal and informal Biological Science education and contribute to achieving quality education for all (Objective # 4 of the Agenda for Sustainable Development 2030, UNESCO) beyond geographical and cultural barriers.This pilot project will be implemented by an interinstitutional, multidisciplinary and international team. It will capitalize on confluent groups´ previous experience and complementary strengths in Science didactics (particularly biology), engineering, arts, virtual reality, fabrication, as well as open hardware and open software culture.The experimental approach corresponds to a quasi-experimental pre-test/post-test design with control groups, and formative and summative evaluation.As a result of the implementation of this educational innovation we expect to contribute to the improvement of Biological Science Teachers students´ CT skills and promote their active involvement in practical activities that should enhance their professional activity.

Background: Early intervention in workers diagnosed with mental disorders is associated with a lower incidence of relapse and shorter sick leave. However, no studies have been carried out on the effect of early intervention using an evidence-based therapy, Cognitive Behavioral Therapy (CBT), on people with sick leave.Aims: The objectives of the present study are to study whether the type of intervention (early or late) will affect the total duration of the sick leave, the partial duration of the sick leave, the duration of the psychotherapy and the time until return to work after the psychotherapy ends. The sample was composed of 167 participants who were on sick leave for adjustment disorders, anxiety disorders or depressive disorder.Results: The participants who had early intervention with CBT had a significantly shorter duration of total sick leave and partial sick leave, and a shorter time until returning to work after the psychotherapy ended than those who had late intervention. There were no statistically differences in the duration or efficacy of the psychotherapy.Conclusion: We can suggest that providing early access to CBT significantly reduces the length of sick leave in patients with mental disorders.

Background Angiogenesis is essential for tumor growth and formation of metastasis. VEGF is the most potent agiogenic citokine. The aim was to investigate the predictive value of postoperative VEGF serum concentration in patients undergoing surgery for colon cancer. Methods Consecutive patients with colon cancer undergoing surgery with curative intent were included. VEGF was measured in serum at 48 h and postoperative day 4. Cox proportional hazards model was used to estimate its contribution with prognosis. Results A total of 94 patients were included. On multivariate analysis VEGF on postoperative day 4 (HR: 1.05; p = 0.011) was independent prognostic factor of decreased DFS and OS. Five-year DFS (57.7% vs. 85%; p = 0.001) and OS (93% vs. 72%; p = 0.005) were significantly lower in patients with postoperative serum VEGF greater than 370 pg/dl. Conclusion Postoperative VEGF serum concentration was an independent predictor of recurrence. These results must be verified in a prospective independent testing cohort.

Effects of altering the properties of an active site in an enzymatic homogeneous catalyst have been extensively reported. However, the possibility of increasing the number of such sites, as commonly done in heterogeneous catalytic materials, remains unexplored, particularly because those have to accommodate appropriate residues in specific configurations. This possibility was investigated by using a serine ester hydrolase as the target enzyme. By using the Protein Energy Landscape Exploration software, which maps ligand diffusion and binding, we found a potential binding pocket capable of holding an extra catalytic triad and oxyanion hole contacts. By introducing two mutations, this binding pocket became a catalytic site. Its substrate specificity, substrate preference, and catalytic activity were different from those of the native site of the wild type ester hydrolase and other hydrolases, due to the differences in the active site architecture. Converting the binding pocket into an extra catalytic active site was proven to be a successful approach to create a serine ester hydrolase with two functional reactive groups. Our results illustrate the accuracy and predictive nature of modern modeling techniques, opening novel catalytic opportunities coming from the presence of different catalytic environments in single enzymes. This project received funding from the European Union’s Horizon 2020 research and innovation program [Blue Growth: Unlocking the potential of Seas and Oceans] under grant agreement no. [634486] (project acronym INMARE). This research was also supported by the grants PCIN-2014-107 (within ERA NET IB2 grant nr. ERA-IB-14-030 - MetaCat), PCIN-2017-078 (within the ERA-MarineBiotech grant ProBone), BIO2014-54494-R, CTQ2016-79138-R and BIO2017-85522-R from the Spanish Ministry of Economy, Industry and Competitiveness. P.N.G. gratefully acknowledges funding from the UK Biotechnology and Biological Sciences Research Council (grant no. BB/M029085/1). R.B. and P.N.G. acknowledge the support of the Supercomputing Wales project, which is part-funded by the European Regional De-velopment Fund (ERDF) via Welsh Government. P.N.G. acknowledges the support of the Centre of Environmental Biotechnology Project funded by the European Regional Development Fund (ERDF) through Welsh Government. The authors gratefully acknowledge financial support provided by the European Regional Development Fund (ERDF). The MALDI-TOF/TOF analysis was performed in the proteomics facility of the Spanish National Center for Biotechnology (CNB-CSIC) that belongs to ProteoRed, PRB2-ISCIII, sup-ported by grant PT13/0001.

Enzyme engineering has allowed not only the de novo creation of active sites catalysing known biological reactions with rates close to diffusion limits, but also the generation of abiological sites performing new-to-nature reactions. However, the catalytic advantages of engineering multiple active sites into a single protein scaffold are yet to be established. Here, we report on proteins with two active sites of biological and/or abiological origin, for improved natural and non-natural catalysis. The approach increased the catalytic properties, such as enzyme efficiency, substrate scope, stereoselectivity and optimal temperature window, of an esterase containing two biological sites. Then, one of the active sites was metamorphosed into a metal-complex chemocatalytic site for oxidation and Friedel–Crafts alkylation reactions, facilitating synergistic chemo- and biocatalysis in a single protein. The transformations of 1-naphthyl acetate into 1,4-naphthoquinone (conversion approx. 100%) and vinyl crotonate and benzene into 3-phenylbutyric acid (≥83%; e.e. >99.9%) were achieved in one pot with this artificial multifunctional metalloenzyme., This work was funded by grant ‘INMARE’ from the European Union’s Horizon 2020 (grant agreement no. 634486), grants PCIN-2017-078 (within the Marine Biotechnology ERA-NET), CTQ2016-79138-R, BIO2016-76601-C3-1-R, BIO2016-76601-C3-3-R, BIO2017-85522-R, RTI2018-095166-B-I00 and RTI2018-095090-B-100 from the Ministerio de Economía y Competitividad, the Ministerio de Ciencia, Innovación y Universidades (MCIU), the Agencia Estatal de Investigación (AEI), the Fondo Europeo de Desarrollo Regional (FEDER) and the European Union (EU). P.N.G. and R.B. acknowledge the support of the UK Biotechnology and Biological Sciences Research Council (BBSRC; grant No. BB/M029085/1) and the Centre of Environmental Biotechnology Project and the Supercomputing Wales project, which are partly funded by the European Regional Development Fund (ERDF) through the Welsh Government. The authors gratefully acknowledge the financial support provided by the ERDF. C.C. thanks the Ministerio de Economía y Competitividad and FEDER for a Ph.D. fellowship (Grant BES-2015-073829). J.L.G.-A. thanks the support of the Spanish Ministry of Education, Culture and Sport through the National Program FPU (FPU17/00044). I.C.-R. thanks the Regional Government of Madrid for a fellowship (PEJ_BIO_AI_1201). The authors would like to acknowledge S. Ciordia and M. C. Mena for MALDI-TOF/TOF analysis. We thank the staff of both the European Synchrotron Radiation Facility (ESRF, Grenoble, France), for providing access and technical assistance at beamline ID30A-1/MASSIf-1, and the Synchrotron Radiation Source at Alba (Barcelona, Spain), for assistance at BL13-XALOC beamline. The authors would also like to acknowledge M. J. Vicente and M. A. Pascual at the Servicio Interdepartamental de Investigación (SIDI) of the Autonomous University of Madrid for the ESI-MS analyses.

A global census of marine microbial life has been underway over the past several decades. During this period, there have been scientific breakthroughs in estimating microbial diversity and understanding microbial functioning and ecology. It is estimated that the ocean, covering 71% of the earth's surface with its estimated volume of about 2 × 1018 m3 and an average depth of 3800 m, hosts the largest population of microbes on Earth. More than 2 million eukaryotic and prokaryotic species are thought to thrive both in the ocean and on its surface. Prokaryotic cell abundances can reach densities of up to 1012 cells per millilitre, exceeding eukaryotic densities of around 106 cells per millilitre of seawater. Besides their large numbers and abundance, marine microbial assemblages and their organic catalysts (enzymes) have a largely underestimated value for their use in the development of industrial products and processes. In this perspective article, we identified critical gaps in knowledge and technology to fast-track this development. We provided a general overview of the presumptive microbial assemblages in oceans, and an estimation of what is known and the enzymes that have been currently retrieved. We also discussed recent advances made in this area by the collaborative European Horizon 2020 project ‘INMARE’., The success of using marine resources for the development of industrial products and processes depends on the manner they are screened, produced and tested on a larger scale.

Periwinkles of the family Littorinidae (Children, 1834) are common members of seashore littoral communities worldwide. Although the family is composed of more than 200 species belonging to 18 genera, chromosome numbers have been described in only eleven of them. A molecular cytogenetic analysis of nine periwinkle species, the rough periwinkles Littorina arcana, L. saxatilis, and L. compressa, the flat periwinkles L. obtusata and L. fabalis, the common periwinkle L. littorea, the mangrove periwinkle Littoraria angulifera, the beaded periwinkle Cenchritis muricatus, and the small periwinkle Melarhaphe neritoides was performed. All species showed diploid chromosome numbers of 2n = 34, and karyotypes were mostly composed of metacentric and submetacentric chromosome pairs. None of the periwinkle species showed chromosomal differences between male and female specimens. The chromosomal mapping of major and minor rDNA and H3 histone gene clusters by fluorescent in situ hybridization demonstrated that the patterns of distribution of these DNA sequences were conserved among closely related species and differed among less related ones. All signals occupied separated loci on different chromosome pairs without any evidence of co-localization in any of the species.

An important goal of mental health specialists is to improve the quality of life and the adaptation of people with common mental disorders on sick leave. Meaning in life is a protective factor for people adjusting to distress and negative events. This study explores the buffering role of meaning in life in the relationship between clinical anxiety or negative affect and maladjustment in participants with common mental disorders on sick leave. The sample was 167 participants with Adjustment, Anxiety and, Depressive Disorders; n = 115 were women and n = 52 men. Participants’ mean age M = 42.16 (SD = 9.91) years. We performed zero-order correlations and hierarchical regression analyses. Meaning in life moderated and buffered the association between clinical anxiety or negative affect and global maladjustment to daily life. These findings suggest that Meaning in life is a relevant variable in the adjustment of common mental disorders on sick leave.

This work received the honour of being the Cover of the ACS Chemical Neuroscience in its issue of May 2015, 6-Methoxy-1,2,3,4-tetrahydro-β-carboline (pinoline) and N-acetyl-5-methoxytryptamine (melatonin) are both structurally related to 5-hydroxytryptamine (serotonin). Here we describe the design, synthesis, and characterization of a series of melatonin rigid analogues resulting from the hybridization of both pinoline and melatonin structures. The pharmacological evaluation of melatonin-pinoline hybrids comprises serotonergic and melatonergic receptors, metabolic enzymes (monoamine oxidases), antioxidant potential, the in vitro blood-brain barrier permeability, and neurogenic studies. Pinoline at trace concentrations and 2-acetyl-6-methoxy-1,2,3,4-tetrahydro-β-carboline (2) were able to stimulate early neurogenesis and neuronal maturation in an in vitro model of neural stem cells isolated from the adult rat subventricular zone. Such effects are presumably mediated via serotonergic and melatonergic stimulation, respectively., This work was supported by the Spanish Ministry of Economy and Competitiveness (Project SAF2012-31035), the Fundación de Investigacion Mediica Mutua Madrileña Automovilistica (AP103952012), and the CSIC (PIE-201280E074). M.F.R. had a JAE-Predoctoral Contract (Grant JAE-Pre-2009-106) from the Program “Junta para la Ampliación de Estudios” cofinanced by the CSIC and the European Social Fund.

Background Anastomotic leakage is associated with higher rates of recurrence after surgery for colorectal cancer. However, the mechanisms responsible are unknown. The aim was to investigate the inflammatory and angiogenic responses in patients undergoing surgery for colorectal cancer who had postoperative intra-abdominal infection, and to compare the results with patients without complications. Methods Consecutive patients undergoing surgery for colorectal cancer with curative intent were included. Patients who had an anastomotic leak or intra-abdominal abscess were included in the infection group and matched with patients who had an uncomplicated postoperative course. IL-6 and VEGF were measured in serum and peritoneal fluid. Results Serum concentration of IL-6 was higher in the infection group (n = 30) compared with the control group (n = 30) on day 4 (infection: 42.3 [27.6–1473.2] versus control: 0.6 [0.6–17.1] pg/ml; p = 0.008). IL-6 in peritoneal fluid was higher in the infection group at 48 h and day 4 (infection: 1000.2 [995.4–1574.0] versus control: 90.3 [35.2.6–106.1] pg/ml; p = 0.001). Serum VEGF was higher in the infection group on day 4 (infection: 1128.6 [427.3–10000.0] versus control: 438.3 [214.1–677.6] pg/ml; p = 0.001). Peritoneal VEGF concentration was higher in the infection group at 48 h and day 4 (infection: 10000.0 [2563.0–10000.0] versus control: 477.8 [313.5–814.4] pg/ml; p = 0.001). Two-year recurrence rate was higher in patients with infection (infection: 30% versus control: 4%; p = 0.001). Conclusions Intra-abdominal infection increases IL-6 and VEGF after surgery for colorectal cancer. Amplification of inflammation and angiogenesis might be one of the mechanisms responsible for the higher recurrence rate observed in patients with anastomotic leakage or intra-abdominal abscess.

Objectives To determine the incidence of incisional hernia (IH) in the extraction incision (EI) in colorectal resection for cancer. To analyze whether the location of the incision has any relationship with the incidence of hernias and whether mesh could be useful for prevention in high-risk patients. Methods Retrospective review of the colon and rectal surgery database from January 2015 to December 2016. Data were classified into 2 groups, transverse (TI) and midline incision (MI), and the latter was divided into 2 subgroups (mesh [MIM] and suture [MIS]). Patients were classified using the HERNIAscore. Hernias were diagnosed by clinical and/or CT examination. Results A total of 182 out of 210 surgical patients were included. After a median follow-up of 13.0 months, 39 IH (21.9%) were detected, 23 of which (13.4%) were in the EI; their frequency was lower in the TI group (3.4%) and in the MIM group (5.9%) than in the MIS group (29.5%; P=.007). The probability of developing IH in the MIS group showed an OR=11.7 (95% CI: 3.3–42.0) compared to the TI group and 4.3 (IC 95%: 1.1–16.3) vs the MIM group. Conclusions The location of the incision is relevant to avoid incisional hernias. Transverse incisions should be used as the first option. When a midline incision is needed, a prophylactic mesh could be considered in high risk patients because it is safe and associated with low morbidity.

RESUMEN Introducción y objetivo: el tatuaje endoscópico preoperatorio es un procedimiento eficaz que permite la localización intraoperatoria de neoplasias de pequeño tamaño. Sin embargo, actualmente no existen unos criterios definidos sobre las indicaciones del tatuaje endoscópico de estas neoplasias en el momento del diagnóstico. El objetivo es establecer unos criterios endoscópicos para seleccionar los pacientes que precisarán un tatuaje. Material y métodos: estudio ambispectivo de pacientes intervenidos de neoplasia colorrectal por laparoscopia en los que se realizó tatuaje endoscópico en el periodo (2007-2013 y 2016-2017). De acuerdo con la descripción endoscópica de las neoplasias se clasificaron en: lesiones polipoideas, neoplasias que ocupan < 50% o ≥ 50% de la luz intestinal y neoplasias estenosantes. Resultados: se realizó tatuaje de la lesión en 120 pacientes y en 114 (95%) se identificó el mismo durante la cirugía. La mayor parte de las neoplasias descritas como polipoideas y neoplasias que ocupaban < 50% de la luz intestinal no se visualizaban en la cirugía y por tanto precisaban el tatuaje (33 de 42 y 18 de 26 respectivamente, p = 0.0001, 2). En cambio, aquellas lesiones estenosantes o bien neoplasias que ocupaban ≥ 50% de la luz intestinal se identificaban mayoritariamente en la cirugía (15 de 15 y 36 de 37 respectivamente, p = 0.0001, 2) sin necesidad de tatuaje. En conjunto la identificación de las neoplasias según los criterios establecidos fue del 98%. Conclusiones: estos resultados sugieren que es posible establecer unos criterios endoscópicos que permitan realizar un tatuaje selectivo durante la endoscopia diagnóstica manteniendo el éxito del mismo. ABSTRACT Background and aim: Preoperative endoscopic tattooing is an effective procedure to identify small intraoperative neoplasms. However, there are no defined criteria with regard to the indications for endoscopic tattooing of these lesions at the time of diagnosis. The aim of this study was to establish endoscopic criteria that allow the selection of patients who will need a tattoo during the diagnostic colonoscopy. Methods: An ambispective study of patients undergoing laparoscopy due to a colorectal neoplasia who underwent endoscopic tattooing during the period from 2007-2013 and 2016-2017. According to the endoscopic description of the neoplasms, the classification was polypoid lesions, neoplasms occupying < 50% or ≥ 50% of the intestinal lumen and stenosing neoplasias. Results: Tattooing of the lesion was performed in 120 patients and the same lesions were identified during surgery in 114 (95%) cases. Most of the neoplasias described as polypoids and neoplasias that occupied < 50% of the intestinal lumen were not visualized during surgery and therefore required a tattoo (33 of 42 and 18 of 26 respectively, p = 0.0001, 2). On the other hand, stenosing lesions or neoplasias occupying ≥ 50% of the intestinal lumen were mostly identified during surgery (15 of 15 and 36 of 37 respectively, p = 0.0001, 2) without the need for a tattoo. Overall, the identification of neoplasms according to established criteria was 98%. Conclusion: These results suggest that it is possible to establish endoscopic criteria that allow a successful selective tattooing during diagnostic endoscopy.

RESUMEN Introducción y objetivo: el tatuaje endoscópico preoperatorio es un procedimiento eficaz que permite la localización intraoperatoria de neoplasias de pequeño tamaño. Sin embargo, actualmente no existen unos criterios definidos sobre las indicaciones del tatuaje endoscópico de estas neoplasias en el momento del diagnóstico. El objetivo es establecer unos criterios endoscópicos para seleccionar los pacientes que precisarán un tatuaje. Material y métodos: estudio ambispectivo de pacientes intervenidos de neoplasia colorrectal por laparoscopia en los que se realizó tatuaje endoscópico en el periodo (2007-2013 y 2016-2017). De acuerdo con la descripción endoscópica de las neoplasias se clasificaron en: lesiones polipoideas, neoplasias que ocupan < 50% o ≥ 50% de la luz intestinal y neoplasias estenosantes. Resultados: se realizó tatuaje de la lesión en 120 pacientes y en 114 (95%) se identificó el mismo durante la cirugía. La mayor parte de las neoplasias descritas como polipoideas y neoplasias que ocupaban < 50% de la luz intestinal no se visualizaban en la cirugía y por tanto precisaban el tatuaje (33 de 42 y 18 de 26 respectivamente, p = 0.0001, 2). En cambio, aquellas lesiones estenosantes o bien neoplasias que ocupaban ≥ 50% de la luz intestinal se identificaban mayoritariamente en la cirugía (15 de 15 y 36 de 37 respectivamente, p = 0.0001, 2) sin necesidad de tatuaje. En conjunto la identificación de las neoplasias según los criterios establecidos fue del 98%. Conclusiones: estos resultados sugieren que es posible establecer unos criterios endoscópicos que permitan realizar un tatuaje selectivo durante la endoscopia diagnóstica manteniendo el éxito del mismo.

Peritoneal infection after colorectal cancer surgery is associated with a higher rate of tumor relapse. We have recently proposed that soluble inflammatory factors released in response to a postoperative infection enhance tumor progression features in residual tumor cells. In an effort to set up models to study the mechanisms of residual tumor cell activation during surgery-associated inflammation, we have analyzed the phenotypic response of colon cancer cell lines to the paracrine effects of THP-1 and U937 differentiated human macrophages, which release an inflammatory medium characteristic of an innate immune response. The exposure of the colon cancer cell lines HT-29 and SW620 to conditioned media isolated from differentiated THP-1 and U937 macrophages induced a mesenchymal-like phenotypic shift, involving the activation of in vitro invasiveness. The inflammatory media activated the β-catenin/TCF4 transcriptional pathway and induced the expression of several mesenchymal (e.g., FN1 and VIM) and TCF4 target genes (e.g., MMP7, PTGS2, MET, and CCD1). Similarly, differential expression of some transcription factors involved in epithelial-to-mesenchymal transitions (i.e. ZEB1, SNAI1, and SNAI2) was variably observed in the colon cancer cell lines when exposed to the inflammatory media. THP-1 and U937 macrophages, which displayed characteristics of M1 differentiation, overexpressed some cytokines previously shown to be induced in colorectal cancer patients with increased rates of tumor recurrence associated with postoperative peritoneal infections, thus suggesting their pro-tumoral character. Therefore, the environment created by inflammatory M1 macrophages enhances features of epithelial-to-mesenchymal transition, and may be useful as a model to characterize pro-inflammatory cytokines as putative biomarkers of tumor recurrence risk.