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Background and aims: Leptin receptor (LEPR) deficiency promotes severe obesity and metabolic disorders. However, the current therapeutic options against this syndrome are scarce. Methods: db/db mice and their wildtypes were systemically treated with neuronal-targeted small extracellular vesicles (sEVs) harboring a plasmid encoding a dominant negative mutant of AMP-activated protein kinase alpha 1 (AMPKα1-DN) driven by steroidogenic factor 1 (SF1) promoter; this approach allowed to modulate AMPK activity, specifically in SF1 cells of the ventromedial nucleus of the hypothalamus (VMH). Animals were metabolically phenotyped. Results: db/db mice intravenously injected with SF1-AMPKα1-DN loaded sEVs showed a marked feeding-independent weight loss and decreased adiposity, associated with increased sympathetic tone, brown adipose tissue (BAT) thermogenesis and browning of white adipose tissue (WAT).Conclusion: Overall, this evidence indicates that specific modulation of hypothalamic AMPK using a sEV-based technology may be a suitable strategy against genetic forms of obesity, such as LEPR deficiency Ministerio de Ciencia y Universidades co-funded by the FEDER Program of EU (CD: BFU2017-87721; RN: RTI2018-099413-B-I00 and RED2018-102379-T; ML: RTI2018-101840-B-I00, PID2021-128145NB-I00 and PDC2022-133958-I00). “la Caixa” Foundation (ID100010434), under the agreement LCF/PR/HR19/52160022 (ML); EuroNanoMed III (RA & ML: EURONANOMED2019-050-ENAMEP); European Research Council (RN: ERC Synergy Grant-2019-WATCH-810331) SI

Selenium (Se) is an essential micronutrient with several functions in cellular and molecular anticancer processes. There is evidence that Se depending on its chemical form and the dosage use could act as a modulator in some anticancer mechanisms. However, the metabolism of organic and inorganic forms of dietary selenium converges on the main pathways. Different selenocompounds have been reported to have crucial roles as chemopreventive agents, such as antioxidant activity, activation of apoptotic pathways, selective cytotoxicity, antiangiogenic effect, and cell cycle modulation. Nowadays, great interest has arisen to find therapies that could enhance the antitumor effects of different Se sources. Herein, different studies are reported related to the effects of combinatorial therapies, where Se is used in combination with proteins, polysaccharides, chemotherapeutic agents or as nanoparticles. Another important factor is the presence of single nucleotide polymorphisms in genes related to Se metabolism or selenoprotein synthesis which could prevent cancer. These studies and mechanisms show promising results in cancer therapies. This review aims to compile studies that have demonstrated the anticancer effects of Se at molecular levels and its potential to be used as chemopreventive and in cancer treatment.

[EN] Celiac disease is a complex and autoimmune disorder caused by the ingestion of gluten affecting almost 1% of global population. Nowadays an effective treatment does not exist, and the only way to manage the disease is the removal of gluten from the diet. Owing the key role played by gluten, clear and regulated labelling of foodstuff and smart methods for gluten detection are needed to fight frauds on food industry and to avoid the involuntary ingestion of this protein by celiac patients. On that scope, the development of a novel detection system of gluten is here presented. The sensor consists of nanoporous anodic alumina films loaded with a fluorescent dye and capped with an aptamer that recognizes gliadin (gluten's soluble proteins). In the presence of gliadin, aptamer sequences displace from the surface of anodic alumina resulting in pore opening and dye delivery. The dispositive shows a limit of detection (LOD) of 100 mu g kg(-1) of gliadin, good selectivity and a detection time of approximately 60 min. Moreover, the sensor is validated in real food samples. This novel probe allows fast gluten detection through a simple signalling process with potential use for food control. (C) 2020 Elsevier B.V. All rights reserved., The authors want to thank the Spanish Government RTI2018-100910-B-C41 (MCUI/AEI/FEDER, UE) and the Generalitat Valenciana (project PROMETEO/2018/024) for support. L.P. thanks to Ministerio de Economia, Industria y Competitividad for his FPI grant. S.S.-F. thanks CIBER and ISCIII for her Sara Borrell contract. The authors also thank the Electron Microscopy Service at the UPV for support.

Background Intravenous drug administration is associated with potential complications, such as phlebitis. The physiochemical characteristics of the infusate play a very important role in some of these problems. Aim The aim of this study was to standardize the dilutions of intravenous drugs most commonly used in hospitalized adult patients and to characterize their pH, osmolarity and cytotoxic nature to better guide the selection of the most appropriate vascular access. Methods The project was conducted in three phases: (i) standardization of intravenous therapy, which was conducted using a modified double-round Delphi method; (ii) characterization of the dilutions agreed on in the previous phase by means of determining the osmolarity and pH of each of the agreed concentrations, and recording the vesicant nature based on the information in literature; and (iii) algorithm proposal for selecting the most appropriate vascular access, taking into account the information gathered in the previous phases. Results In total, 112 drugs were standardized and 307 different admixtures were assessed for pH, osmolarity and vesicant nature. Of these, 123 admixtures (40%), had osmolarity values >600 mOsm/L, pH < 4 or > 9, or were classified as vesicants. In these cases, selection of the most suitable route of infusion and vascular access device is crucial to minimize the risk of phlebitis-type complications. Conclusions Increasing safety of intravenous therapy should be a priority in the healthcare settings. Knowing the characteristics of drugs to assess the risk involved in their administration related to their physicochemical nature may be useful to guide decision making regarding the most appropriate vascular access and devices.

Background Familial adenomatous polyposis is described as one of the common two types of genetic disorders: APC and MUTYH gene associated polyposis syndrome and the clinical differences between the two can sometimes be unclear. Materials and methods A retrospective analysis and comparison was made of clinical, surgical, and histological criteria, mutation types and the long-term results of patients who underwent genetic analysis which resulted in the diagnosis of familial adenomatous polyposis between 1984 and 2018. Results Of the total 71 patients included in the study, 14 were identified with the MUTYH gene, and 57 with the APC mutation. In patients with the APC mutation, 63% had duodenal adenoma, 61% gastric polyp and 54% had desmoid tumor. Of the patients with the MUTYH mutation, 21% had duodenal adenoma and 21% were diagnosed with gastric polyps. In 21% of the patients with APC mutation, the polyp count was 100 polyps in the colon No statistical difference was determined between the groups in respect of the proportion of patients with >100 polyps. Conclusion The pre-operative genetic testing of patients with polyposis coli will be useful in determining the future clinical outcome and helpful in guiding an informed decision as to whether to apply surgical treatment. It is useful to determine the colonic and extra-colonic involvement of genetic mutation diseases in patients with familial adenomatous polyposis.

Resumen Antecedentes La poliposis adenomatosa familiar (PAF) es una patologia hereditaria, caracterizada por la existencia de polipos y cancer en el colon. La PAF puede ser consencuencia de dos trastornos geneticos: El gen adenomatous polyposis coli (APC) o el gen mutacion Y homologo (MUTYH). Las diferencias clinicas y fenotipicas entre las dos alteraciones geneticas no estan claramente establecidas. Materiales y metodos Se realizo un analisis restrospectivo de las manifestaciones clinicas, criterios quirurgicos, caracteristicas histologicas, tipo de mutacion y resultados a largo plazo de pacientes diagnosticados mediante analisis geneticos de poliposis adenomatosa familiar entre 1984 y 2018. Resultados De un total de 71 pacientes incluidos en el estudio, en 14 de ellos se identifico mutacion del gen MUTYH y en 57, mutacion del gen APC. A 60 pacientes se les realizo tratamiento quirurgico, a la mitad de ellos se les practico proctocolectomia y a la otra mitad, colectomia total. En pacientes con la mutacion APC, el 63% presento adenomas duodenales; el 61%, polipos gastricos y el 54% tumor desmoide. De los pacientes con la mutacion del gen MUTYH, el 21% presento adenomas duodenales y al 21% se le diagnostico polipos gastricos. En el 21% de los pacientes con mutacion del gen APC, el numero de polipos fue inferior a 100 y en el 64% de los pacientes que presentaron mutacion del gen MUTYH se observaron mas de 100 polipos en el colon. No existio diferencias estadisticamente significativas entre lo grupos respecto a la proporcion de pacientes con mas de 100 polipos. Conclusion Es importante valorar la afectacion colonica y la extracolonica en pacientes con mutaciones geneticas asociadas a la PAF.

Multiwalled carbon nanotubes (MWCNTs) are tubular carbon structures that are able to enter cells through holes in the plasma membrane and produce changes in gene expression. In this work, we compared the functionality of carbon nanotubes with the electroporation that perforates membranes, in Brassica oleracea var. Italica (broccoli) root protoplasts. For this, we combined those treatments with control conditions and abiotic stress (salinity) in order to elucidate if the response is related to conditions optimal for the plant. The measurement of the osmotic water permeability (Pf), mineral concentrations and expression levels of aquaporins (PIP1s and PIP2s) revealed that the physiological action of the nanotubes was similar to that achieved with electroporation for both Pf and the concentrations of nutrients in the protoplasts. On the other hand, PIP1s and PIP2s expression was increased in the protoplasts receiving the control plus MWCNTs treatment but not in those treated with electroporation. This opens new and interesting lines, as it shows that nanotubes are able to modulate the expression of aquaporins.

Anal intraepithelial neoplasia (AIN) is a premalignant lesion of anal squamous cell carcinoma. HIV-positive males who have sex with males, are the most affected at-risk population. Cytology and anuscopy are the best accepted methods for its diagnosis, although it is controversial which patients should complete it with a biopsy. Neither which patients should undergo treatment nor which is the best treatment is not well established. With this study, we would like to present our experience in the diagnostic-therapeutic management of AIN in the short term.Retrospective observational study of patients at risk of AIN with altered anal cytology who underwent high-resolution anuscopy with biopsy. After histological confirmation of dysplasia, they started treatment with trichloroacetic acid. Its effectiveness was verified by subsequent cytology. The demographic variables of the sample and the results of both diagnostic and treatment tests were analyzed.The majority were HIV-positive males (104/115) and 50% had sexual relations with other men. We included 115 patients with altered anal cytology, of whom 92% had dysplasia on biopsy. 97% with atypia of uncertain significance on cytology had histological dysplasia. Cytology normalized after treatment in 60% of patients.Early detection of AIN should be routinely considered in known at-risk populations. Any cytological abnormality should be biopsied. Tricholoroacetic acid can be an effective treatment achieving a high percentage of regression, although currently, the information we have is of low level of evidence.

The circumstances arising from the exceptional situation caused by the COVID-19 pandemic have affected all socioeconomic areas in the last two years. The field of Education has not been an exception, and the management of the situation seems to have caused an increase in the level of perceived stress of university students. On this basis, this research that aims, first, to analyze the student’s level of perceived stress during the de-escalation and return to normality period, secondly, to evaluate its relationship with life satisfaction and, finally, to detect the students’ needs in stress management, has been developed. An exploratory-descriptive study of quantitative and cross-sectional nature has been carried out. A total of 222 university students of the Childhood and Primary Education Degrees of the University of Jaén (Spain) have participated. The instruments, Perceived Stress Scale (PSS) and Satisfaction with Life Scale (SWLS) have been used. The results show a moderate and partial relationship among the dimensions of the used instruments. The obtained coefficients of determination are, Academic Performance (r² = 0.019) and Life Satisfaction (r² = 0.402), with a mean square error (SRMR) of 0.079. These findings show the need to develop actions within the university training program in effective stress management strategies.

The epidemics of obesity and type 2 diabetes have led to intensive investigation of the underlying mechanisms of these diseases and their main complications such as cardiovascular diseases and non-alcoholic fatty liver disease. This search has contributed to better understand how organs and tissues communicate with each other in the so-called inter-organ crosstalk. Adipose tissue, the liver, or skeletal muscle can actively release secreted factors termed "organokines" which can interact with other distant targets in complex networks. More recently, other novel mediators of inter-organ crosstalk such as extracellular vesicles and their non-traditional cargoes as miRNAs and lncRNAs are gaining importance and represent potential therapeutic targets. In the present chapter we summarize some of the current knowledge on inter-organ communication with a focus on adipose tissue-released factors and their modulation on other organs and tissues like pancreas, liver, skeletal muscle, the cardiovascular system, and the gut in the context of obesity and its progression to insulin resistance. We also provide a perspective on mediators of inter-organ crosstalk as potential therapeutic targets.

This study aimed to highlight the relationship between diet, animal performance and mucosal adherent gut microbiota (anterior intestine) in fish fed plant-based diets supplemented with an egg white hydrolysate (EWH) with antioxidant and anti-obesogenic activity in obese rats. The feeding trial with juveniles of gilthead sea bream (Sparus aurata) lasted 8 weeks. Fish were fed near to visual satiety with a fish meal (FM)/fish oil (FO) based diet (CTRL) or a plant-based diet with/without EWH supplementation. Specific growth rate decreased gradually from 2.16% in CTRL fish to 1.88% in EWH fish due to a reduced feed intake, and a slight impairment of feed conversion ratio. Plant-based diets feeding triggered a hyperplasic inflammation of the anterior intestine regardless of EWH supplementation. However, EWH ameliorated the goblet cell depletion, and the hepatic and intestinal lipid accumulation induced by FM/FO replacement. Illumina sequencing of gut mucosal microbiota yielded a mean of 136,252 reads per sample assigned to 2,117 OTUs at 97% identity threshold. The bacterial diversity was similar in all groups, but a significantly lower richness was found in EWH fish. At the phylum level, Proteobacteria reached the highest proportion in CTRL and EWH fish, whereas Firmicutes were decreased and Actinobacteria increased with the FM/FO replacement. The proportion of Actinobacteria was restored by dietary EWH supplementation, which also triggered a highest amount of Bacteroidetes and Spirochaetes. At a closer look, a widespread presence of Lactobacillales among groups was found. Otherwise, polysaccharide hydrolases secretors represented by Corynebacterium and Nocardioides were increased by the FM/FO replacement, whereas the mucin-degrading Streptococcus was only raised in fish fed the plant-based diet without EWH. In addition, in EWH fish, a higher abundance of Propionibacterium was related to an increased concentration of intestinal propionate. The antagonism of gut health-promoting propionate with cholesterol could explain the inferred underrepresentation of primary bile acid biosynthesis and steroid degradation pathways in the EWH fish microbiota. Altogether, these results reinforce the central role of gut microbiota in the regulation of host metabolism and lipid metabolism in particular, suggesting a role of the bioactive EWH peptides as an anti-obesity and/or satiety factor in fish., This work was supported by the EU H2020 Research Innovation Program under the TNA Program (project AE150009) at IATS-CSIC Research Infrastructure within AQUAEXCEL2020 Project (652831). This output reflects only the author’s view and the European Union cannot be held responsible for any use that may be made of the information contained herein. Additional funding was obtained by a Spanish MICINN project (Bream-AquaINTECH and RTI2018–094128-B-I00). MCP was funded by a Ramón y Cajal Postdoctoral Research Fellowship [RYC2018-024049-I/AEI/10.13039/501100011033 co-funded by the European Social Fund (ESF) and ACOND/2020 Generalitat Valenciana].

Proton-nuclear-magnetic-resonance-spectroscopy (

Lactococcus lactis contains a large number of key enzymes responsible for the formation of volatile compounds characteristic of cheese flavour. In the present work we have investigated the expression of genes codifying amino acid converting enzymes (AACE) and the formation of volatile compounds by L. lactis IFPL953 and its mutant lacking the hydroxy acid dehydrogenase PanE (L. lactis IFPL953ΔpanE). The growth in absence of isoleucine was the main induction factor in the expression of araT, bcaT, panE, and kivD. The expression of the gdh gene of L. lactis IFPL953ΔpanE increased remarkably during the stationary growth phase, particularly under isoleucine and valine starvation conditions. L. lactis IFPL953ΔpanE showed an enhanced formation of 2- and 3-methylbutanal and their corresponding alcohols and an increase in the formation of ketones. These findings contribute to a better knowledge of L. lactis AACE regulation and its potential application in cheese flavour formation., This study was funded by Spanish Ministry of Science (Grants AGL2016-75951-R and PCIN-2017-003).

Naranjilla (Solanum quitoense) is an Andean tropical fruit of the solanaceae family with high nutritional value. Its cultivation is gravely affected by the lethal disease produced by Fusarium oxysporum f.sp. quitoense, causing severe economic losses. In this work we studied the possibility of boosting the plants’ natural defenses to the fungus by using plant hormones. Our results show that spraying jasmonic acid or sodium nitroprusside (a NO donor) on naranjilla plantlets before Fusarium inoculation protected them from infection, and elicited strong activation of the corresponding signaling pathway, suggesting a mechanism of defense priming triggered by those compounds.

[EN] Microalgae play a fundamental role in aquatic primary production and the food chain. They are a recognized source of fatty acids and fatty acid-based lipids of potential interest in the preparation of functional health products, biofuels and renewable chemicals. The exploitation of this bioresource requires a fine monitoring of each production stage. The aim of this work is the microalgae degradation follow up after the concentration stage at the end of the production process by voltammetric electronic tongue, impedance spectroscopy and H-1 NMR spectroscopy. Microalgae samples were allowed to progress along time (from 1 to 23 days). At scheduled selected times, voltammetry, impedance spectroscopy and H-1 NMR measurements were performed. Multivariate analysis was carried out on these data by PLSR. A model calculated in a training set was then applied to a set of validation to predict the time of evolution. For the three techniques good results in prediction for the validation set were obtained (R-2/RMSEP of 0.961/1.51, 0.956/1.67 and 0.969/1.25 respectively for impedance, voltammetry and NMR spectroscopy). The three techniques were sensitive to the evolution of the microalgae samples. The detection of metabolical changes in the H-1 NMR spectra is also included. This proof of concept could be the basis for future development of rapid and robust strategies for quality control on microalgae production plants., Authors gratefully acknowledge the financial support of MAT2015-64139-C4-1-R and MAT2015-64139-C4-3-R (MINECO/FEDER, UE) and PROMETEOII/2014/047 projects. BUGGYPOWER, S.L. is acknowledged for the financial support and for providing the samples. U26 Nanbiosis is acknowledged by the NMR measurements.

Producción Científica, Corneal cross-linking (CXL) is a surgical procedure able to modify corneal biomechanics and stabilize keratoconus progression. Although it is known that CXL produces changes in corneal collagen distribution, these are still a topic of discussion. Here we quantitatively compare the corneal stroma architecture between two animal models four weeks after in vivo conventional CXL treatment, with second harmonic generation (SHG) imaging microscopy and the structure tensor (ST). The healing stage and the stroma recovery were also analyzed by means of histological sections. Results show that the CXL effects depend on the initial arrangement of the corneal collagen. While the treatment increases the order in corneas with a low level of initial organization, corneas presenting a fairly regular pattern are hardly affected. Histological samples showed active keratocytes in anterior and middle stroma, what means that the recovery is still in progress. The combination of SHG imaging and the ST is able to objectively discriminate the changes suffered by the collagen arrangement after the CXL treatment, whose effectiveness depends on the initial organization of the collagen fibers within the corneal stroma., Sociedad Española para la Investigación de las Diferencias Individuales (grant FIS2016-76163-R), Fundación Séneca (grant 19897/GERM/15)

Nidogen-2 is a basement membrane (BM) glycoprotein that could be a key to understanding why defects in BM regeneration occur after severe trauma to the cornea. We monitored the location and expression of nidogen-2 during corneal repair after alkali burn in rabbits. In rabbits that received both general and ocular topical anaesthesia, the central cornea of the left eye was burned by placing an 8-mm diameter filter paper soaked in 0.5 N NaOH for 60 s. Right corneas were used as controls. The eyes were evaluated at 2, 7, 15, and 30 days after burning and analysed by immunohistochemistry for nidogen-2 and α-smooth muscle actin, a myofibroblast marker. Nidogen-2 mRNA expression levels were determined by quantitative real-time polymerase chain reaction. In control corneas, nidogen-2-positive cells were in all epithelial layers, the endothelium, and the anterior and posterior stromal regions. At Day 2 after the alkali burn, the wound area epithelium and the peripheral epithelium were made up of only 1 to 2 cell layers, all of them nidogen-2 positive. At Day 7 in the wound area, the epithelium consisted of two cell layers, and the basally located cells were mostly nidogen-2 positive. The greatest change was observed at Day 30. At this time, the ulcer prevalence in the alkali-burned corneas was approximately 50% and the central epithelial defects remained. In unepithelialized corneas, frequent epithelial detachments were present, in which almost of the epithelial cells were nidogen-2 negative. The injured stroma was repopulated by activated stromal cells that synthesized nidogen-2. The nidogen-2 was retained in the newly secreted, but disordered, matrix produced mainly by the myofibroblasts localized in the stroma at 7, 15, and 30 days after burning. Thus, even though nidogen-2 was present, it was unable to contribute to the effective regeneration of the BM.

Resumen Objetivo Evidenciar la eficacia de la acupuntura en la ansiedad ante los examenes, en alumnos universitarios. Diseno Estudio experimental sin asignacion aleatoria. Emplazamiento Universidad CEU Cardenal Herrera, Elche, Alicante. Participantes Estudiantes universitario sanos de edad ≥ 18 anos: 23 sujetos en el grupo de intervencion y 21 en el grupo control. Intervenciones El estudio consistio en 10 sesiones de acupuntura (2 por semana) de 30 min cada una, durante las 5 semanas anteriores a los examenes. Al grupo control se le paso el Inventario de Ansiedad de Beck (BAI, Beck Anxiety Inventory) en los mismos tiempos (T1, T2 y T3) en que se realizo las sesiones al grupo de intervencion. Mediciones principales Para la evaluacion de la ansiedad se ha utilizado el BAI, que evalua sobre todo sintomas fisiologicos: 14 de sus 21 items se refieren a sintomas fisiologicos, 4 evaluan aspectos cognitivos y 3 aspectos afectivos. El cuestionario consta de 21 preguntas, con un rango de puntuacion entre 0 y 63. Los puntos de corte son: ansiedad muy baja (0-21), ansiedad moderada (22-35) y ansiedad grave (> 35). Si la puntuacion del BAI es igual o mayor de 22 puntos, la ansiedad es probable. Resultados Tras 5 sesiones de tratamiento se confirma que los niveles de ansiedad disminuyen respecto a la puntuacion obtenida en el BAI antes de comenzar el tratamiento. Despues de 10 sesiones, el grupo de intervencion mantiene los niveles de ansiedad y en el grupo control sin tratamiento se disparan los niveles de ansiedad de forma significativa. Conclusiones La acupuntura ha demostrado ser efectiva para reducir los niveles de ansiedad antes de los examenes en estudiantes de fisioterapia.

Many important human diseases, and especially cancer, have been related to the overproduction of 8-oxo-7,8-dihydro-2 '-deoxyguanosine (8-oxo-dG). This molecule is a product of oxidative stress processes over nucleophilic bases in DNA. In this work, an aptasensor for the rapid, selective and accurate detection of this oncomarker is presented. The aptasensor consists of a nanoporous anodic alumina material loaded with a dye and is functionalized with an aptamer-based "molecular gate". In the presence of target 8-oxo-dG, the capping aptamer displaces from the surface due to the high affinity of the analyte with the capping aptamer, thus inducing delivery of the preloaded fluorescent dye. In contrast, in the absence of 8-oxo-dG, a poor payload delivery is accomplished. This aptamer-based nanodevice has great sensitivity for 8-oxo-dG, resulting in a LOD of 1 nM and a detection time of ca. 60 min. Moreover, the aptasensor is able to accurately detect 8-oxo-dG in unmodified urine and serum without pre-concentration treatments. This diagnostic tool is validated in a set of 38 urine and serum samples from patients diagnosed of colorectal cancer and control patients. These samples are also analyzed using a standardized and specific ELISA kit. The aptasensor displays excellent sensitivity (95.83/100%) and specificity (80/100%) for 8-oxo-dG detection in serum and urine samples, respectively. Our results may serve as a basis for the development of generalized fluorogenic diagnostic platforms for the easy diagnosis of cancer in biofluids as well as for monitoring therapeutic treatments and detection of relapses without the use of expensive equipment or trained personnel.

[EN] A nanodevice consisted on nanoporous anodic alumina (NAA) supports functionalized with specific and selective antibody-based gatekeepers for the detection of IDH1-R132H mutant enzyme is here reported. Molecular profile and tissue mutations of the tumours (such as IDH1/IDH2 mutations in gliomas) are a great source of information that already make a difference in terms of prognosis and prediction of response to combined therapy. However, standardized methodologies to determine this mutation are time-consuming and cannot provide information before or during surgical intervention, which significantly limits their utility in terms of intraoperative decisionmaking. To solve this limitation, our sensing system, in the presence of the target IDH1-R132H mutant enzyme triggers the delivery of a fluorescent reporter from the antibody-capped NAA that is easily detected using a standard fluorimeter in less than 1 h. Good capabilities of the sensor in terms of sensitivity (limit of detection as low as 0.01 ng/mL) and selectivity are determined. The biosensor is validated in seventeen human glioma tissue samples, also analysed by standardized methodologies, showing significant differences between IDH1 wild-type and IDH1-R132H mutant gliomas (p < 0.01) with high sensitivity, specificity and accuracy (87.5/88.9/88.2) for IDH1-R132H mutational status classification in human glioma tissues. This new detection system might play an important role in surgical interventions, anticipating mutational status information in gliomas and supporting thus decision making and patient survival., This study was supported by the Spanish Government (project RTI2018-100910-B-C41 MCUI/AEI/FEDER, UE), the Generalitat Valenciana (project PROMETEO/2018/024), the Universitat Politecnica de Valencia-Instituto de Investigacion Sanitaria La Fe (GLIOVIEW-IDH and GLIOVIEWIDH_2.0 project) and CIBER-BBN (NANOPATH AND CANDI-EYE projects). S.S. thanks the Instituto de Salud Carlos III and the European Social Fund for the financial support "Sara Borrell" (CD16/000237). L.P. thanks to Ministerio de Economia, Industria y Competitividad for his FPI grant. We also a knowledge service provided by Biobanco La Fe of Instituto de Investigacion Sanitaria La Fe, Val`encia, Spain.

Intestinal microbiota seems to play a key role in obesity. The impact of the composition and/or functionality of the obesity-associated microbiota have yet to be fully characterized. This work assessed the significance of the taxonomic composition and/or metabolic activity of obese- microbiota by massive 16S rRNA gene sequencing of the fecal microbiome of obese and normoweight individuals. The obese metabolic activity was also assessed by in vitro incubation of obese and normoweight microbiotas in nutritive mediums with different energy content. We found that the microbiome richness and diversity of the two groups did not differ significantly, except for Chao1 index, significantly higher in normoweight individuals. At phylum level, neither the abundance of Firmicutes or Bacteroidetes nor their ratio was associated with the body mass index. Besides, the relative proportions in Collinsella, Clostridium XIVa, and Catenibacterium were significantly enriched in obese participants, while Alistipes, Clostridium sensu stricto, Romboutsia, and Oscillibacter were significantly diminished. In regard to metabolic activity, short-chain fatty acids content was significant higher in obese individuals, with acetate being the most abundant followed by propionate and butyrate. Acetate and butyrate production was also higher when incubating obese microbiota in mediums mimicking diets with different energy content; interestingly, a reduced capability of propionate production was associated to the obese microbiome. In spite of the large interindividual variability, the obese phenotype seems to be defined more by the abundance and/or the absence of distinct communities of microorganism rather than by the presence of a specific population., This work was supported by the Spanish Ministry of Science and Innovation (AGL2016-75951-R and PID2019-106071RB-I00 Grants)., We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). We thank Marta Gil for her technical support.

This chapter covers methodologies currently in use for the selection and analysis of lactic acid bacteria (LAB) strains to be employed as starter cultures in dairy fermentations. The aspects covered are the methods for enumeration and identification of LAB strains and the evaluation of their technological properties for improving processing conditions and product quality during dairy fermentations. The selection of LAB strains based on health benefit properties and the conditions for the production and maintenance of dairy cultures are also addressed. The production of desirable and industrially robust dairy starter cultures can be achieved by choosing the desired phenotypic traits and the knowledge to select for these traits at the genotypic level. The analysis of a large number of strains for selecting specific properties requires high-throughput screening tests to progressively reduce the number of candidates. The advances in omics technologies and analytical instruments have provided the bases to improve the evaluation and selection of LAB strains that are able to restore the unique characteristics of traditional dairy products and to benefit human health., Financial support from Spanish Ministry of Science (Projects AGL2016-75951-R, PCIN-2017-075 and IDI-20190077) and CYTED (Strategic Project 917PTE0537) is acknowledged.

Producción Científica, The organization of the corneal stoma is modified due to different factors, including pathology, surgery or external damage. Here the changes in the organization of the corneal collagen fibers during natural healing after chemical burn are investigated using second harmonic generation (SHG) imaging. Moreover, the structure tensor (ST) was used as an objective tool for morphological analyses at different time points after burn (up to 6 months). Unlike control corneas that showed a regular distribution, the collagen pattern at 1 month of burn presented a non-organized arrangement. SHG signal levels noticeably decreased and individual fibers were hardly visible. Over time, the healing process led to a progressive re-organization of the fibers that could be quantified through the ST. At 6 months, the stroma distribution reached values similar to those of control eyes and a dominant direction of the fibers re-appeared. The present results show that SHG microscopy imaging combined with the ST method is able to objectively monitor the temporal regeneration of the corneal organization after chemical burn. Future implementations of this approach into clinically adapted devices would help to diagnose and quantify corneal changes, not only due to chemical damages, but also as a result of disease or surgical procedures., © 2021 Optical Society of America. Users may use, reuse, and build upon the article, or use the article for text or data mining, so long as such uses are for non-commercial purposes and appropriate attribution is maintained. All other rights are reserved., Ministerio de Sanidad. Instituto de Salud Carlos III (PI15/01906), Fundación Séneca (19897/GERM/15)

Given the rise in cyberbullying among secondary education students and the importance of certain psychological adjustment variables for the comprehension of this type of violent behavior, both in bullies and in victims, the purpose of the present study was to analyse the emotional adjustment of those involved in cyber- and traditional bullying. The adjustment variables studied were self-concept, perceived stress, loneliness, depressive symptomatology, social anxiety, life satisfaction, and emotional intelligence. Using a sample of 1318 adolescents (47% boys), aged between 11 and 17 years, four groups were established to compare victims and cybervictims (uninvolved students, traditional victims, cybervictims, and traditional–cybervictims). The analysis of variance showed that students who performed the same role (bully or victim) in both contexts (at school and online). In particular, those who suffered traditional or cyberbullying or both conjointly presented lower scores in physical and social self-concept, life satisfaction, emotional clarity, and emotion regulation, as well as higher scores in perceived stress, loneliness, depression, and social anxiety. In bullies—traditional, cyberbullying or both simultaneously—higher scores were observed in perceived stress, loneliness, depressive symptomatology, fear of negative assessment, avoidance, and general social anxiety, and lower scores in the dimensions of academic and family self-concept, life satisfaction, emotional clarity, and emotion regulation. In general, the findings indicate that students who were involved in bullying situations, both victims and bullies, presented more damaged emotional profiles than those who are uninvolved, especially students who performed the same role (bully or victim) in both contexts (at school and online). This work is part of the Project “Bullying, cyberbullying and child-to-parent violence in adolescence”—Reference: PSI2015-65683-P [MINECO/ERDF, EU], funded by the Ministry of Economy and Competitiveness of Spain and the European Union through the European Regional Development Fund-FEDER- “One way to make Europe”.

Familial adenomatous polyposis is described as one of the common two types of genetic disorders: APC and MUTYH gene associated polyposis syndrome and the clinical differences between the two can sometimes be unclear.A retrospective analysis and comparison was made of clinical, surgical, and histological criteria, mutation types and the long-term results of patients who underwent genetic analysis which resulted in the diagnosis of Familial Adenomatous Polyposis between 1984 and 2018.Of the total 71 patients included in the study, 14 were identified with the MUTYH gene, and 57 with the APC mutation. In patients with the APC mutation, 63% had duodenal adenoma, 61% gastric polyp and 54% had desmoid tumor. Of the patients with the MUTYH mutation, 21% had duodenal adenoma and 21% were diagnosed with gastric polyps. In 21% of the patients with APC mutation, the polyp count was100, and 64% of those with the MUTYH mutation had100 polyps in the colon No statistical difference was determined between the groups in respect of the proportion of patients with100 polyps.The pre-operative genetic testing of patients with polyposis coli will be useful in determining the future clinical outcome and helpful in guiding an informed decision as to whether to apply surgical treatment. It is useful to determine the colonic and extra-colonic involvement of genetic mutation diseases in patients with Familial adenomatous polyposis.

8 pags., 6 figs., PURPOSE. Photoactivated cornea collagen cross-linking (CXL) increases corneal stiffness by initiating formation of covalent bonds between stromal proteins. Because CXL depends on diffusion to distribute the photoinitiator, a gradient of CXL efficiency with depth is expected that may affect the degree of stromal collagen organization. We used second harmonic generation (SHG) microscopy to investigate the differences in stromal collagen organization in rabbit eyes after corneal CXL in vivo as a function of depth and time after surgery. METHODS. Rabbit corneas were treated in vivo with either riboflavin/UV radiation (UVX) or Rose Bengal/green light (RGX) and evaluated 1 and 2 months after CXL. Collagen fibers were imaged with a custom-built SHG scanning microscope through the central cornea (350 μm depth, 225 × 225 μm en face images). The order coefficient (OC), a metric for collagen organization, and total SHG signal were computed for each depth and compared between treatments. RESULTS. OC values of CXL-treated corneas were larger than untreated corneas by 27% and 20% after 1 month and 38% and 33% after 2 months for the RGX and UVX, respectively. RGX OC values were larger than UVX OC values by 3% and 5% at 1 and 2 months. The SHG signal was higher in CXL corneas than untreated corneas, both at 1 and 2 months after surgery, by 18% and 26% and 1% and 10% for RGX and UVX, respectively. CONCLUSIONS. Increased OC corresponded with increased collagen fiber organization in CXL corneas. Changes in collagen organization parallel reported temporal changes in cornea stiffness after CXL and also, surprisingly, are detected deeper in the stroma than the regions stiffened by collagen cross-links., Supported by the European Research Council Advanced Grant ERC-2011-AdG Ref. 294099, Spanish Government Grants FIS2014-56643 and FIS2017-84753-R, H2020 Research and Innovation Action H2020-ICT-2017 Ref. 779960.

To analyze liver function tests (LFT), weight, metabolic syndrome (MetS) and at risk of meeting MetS criteria (AR-MetS) in children and adolescents on antipsychotics (AP) during a year-long follow-up.Two hundred sixteen patients, AP naïve or quasi-naïve (30 days on AP), were included. Total bilirubin, the enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), weight and other parameters of MetS were measured at baseline, and at 3, 6 and 12 months, while patients remained on the same AP.At baseline, patients (mean age: 14.1 ± 3.1 years; 60.2% male) were on risperidone (N = 143), olanzapine (N = 37), or quetiapine (N = 36), although the sample decreased over time to 67 patients at 12 months (risperidone N = 46, olanzapine N = 10, and quetiapine N = 11). Around 3% of patients had ALT/AST levels that were at least twice the upper limit of normal (ULN) at 3 and 6 months; whereas roughly 19% of patients had ALP levels that were at least twice the ULN in at least one assessment after baseline, but had no clinical symptoms. From baseline to 6 months, significant increases were observed in ALT levels in the whole sample (p = 0.005), whereas ALP increased only in patients on risperidone. Patients showed significant weight gain, and more individuals met criteria for MetS and AR-MetS over time (from baseline: 2.8% and 8.3%, to 1 year: 10.5% and 23.9%, respectively). There was a trend-level group effect in global ALT across time (p = 0.076). Patients with MetS showed higher ALT concentrations (28.9 [18.4-39.4] U/L) than AR-MetS (20.4 [8.5-32.2] U/L), and no-AR-MetS (19.2 [8.4-29.9] U/L).Less than 3% of children and adolescents on AP during 1-year follow-up showed an increase in ALT or AST levels in one or more of the assessments, and none of these increases was of clinical significance. Patients with MetS and AR-MetS increased during this period, and the possible role of ALT levels to monitor these patients deserves further study.

[EN] Colon targeted drug delivery is highly relevant not only to treat colonic local diseases but also for systemic therapies. Mesoporous silica nanoparticles (MSNs) have been demonstrated as useful systems for controlled drug release given their biocompatibility and the possibility of designing gated systems able to release cargo only upon the presence of certain stimuli. We report herein the preparation of three gated MSNs able to deliver their cargo triggered by different stimuli (redox ambient (S1), enzymatic hydrolysis (S2), and a surfactant or being in contact with cell membrane (S3)) and their performance in solution and in vitro with Caco-2 cells. Safranin O dye was used as a model drug to track cargo fate. Studies of cargo permeability in Caco-2 monolayers demonstrated that intracellular safranin O levels were significantly higher in Caco-2 monolayers when using MSNs compared to those of free dye. Internalization assays indicated that S2 nanoparticles were taken up by cells via endocytosis. S2 nanoparticles were selected for in vivo tests in rats. For in vivo assays, capsules were filled with S2 nanoparticles and coated with Eudragit FS 30 D to target colon. The enteric coated capsule containing the MSNs was able to deliver S2 nanoparticles in colon tissue (first step), and then nanoparticles were able to deliver safranin O inside the colonic cells after the enzymatic stimuli (second step). This resulted in high levels of safranin O in colonic tissue combined with low dye levels in plasma and body tissues. The results suggested that this combination of enzyme-responsive gated MSNs and enteric coated capsules may improve the absorption of drugs in colon to treat local diseases with a reduction of systemic effects., The authors acknowledge the financial support from the Spanish Government (Projects MAT2015-64139-C4-1-R, SAF2016-78756 and AGL2015-70235-C2-2-R) and the Generalitat Valenciana (Project GVA/2014/13).

X-ray diffraction has been widely used to characterize the structural properties (strain and structural quality) of semiconductor heterostructures. This work employs hybrid multiple diffraction to analyzer-oriented Zn1−xMgxO layers grown by molecular beam epitaxy on ZnO substrates. In such a low-symmetry material system, additional features appear in symmetric reflection scans, which are described as arising from hybrid multiple diffraction. First, the Bragg conditions necessary for these high-order processes to occur are introduced and applied to explain all the observed satellite reflections, identify the planes that contribute and computea priorithe angles at which they are observed. Furthermore, thanks to this hybrid multiple-diffraction technique, it is possible to determine the layer lattice parameters (in-plane and out-of-plane) in an easy and accurate way by using one single measurement in standard symmetric conditions. The achieved precision is at least as high as that obtained from the combination of symmetric and asymmetric reciprocal space map measurements.

There is limited information that relates the intake of food-derived bioactive peptides and the gut microbiota. We have previously described a pepsin hydrolysate of egg white (EWH) that ameliorates fat accumulation and dyslipidemia, while reducing oxidative stress and inflammation markers in obese Zucker rats. The aim of this study was to associate the beneficial effects of EWH with gut microbiota changes in these animals. Obese Zucker rats received daily 750 mg kg−1 EWH in drinking water for 12 weeks and faeces were analysed for microbial composition and metabolic compounds in comparison with Zucker lean rats and obese controls. EWH supplementation modulated the microbiological characteristics of the obese rats to values similar to those of the lean rats. Specifically, counts of total bacteria, Lactobacillus/Enterococcus and Clostridium leptum in EWH fed obese Zucker rats were more similar to the lean rats than to the obese controls. Besides, feeding the obese Zucker rats with EWH reduced (P < 0.05) the faecal concentration of lactic acid. The physiological benefits of EWH in the improvement of obesity associated complications of Zucker rats could be associated with a more lean-like gut microbiota and a tendency to diminish total short-chain fatty acids (SCFA) production and associated obesity complications. The results warrant the use of pepsin egg white hydrolysate as a bioactive food ingredient., The authors acknowledge funding from the Spanish Ministry for Science and Innovation (AGL2012-35814 and AGL2012-32387) and CSIC (COOPB-20099 and Intramural 201570I028).

Background: The prognosis for patients with colorectal cancer shows variation. The characteristics of colorectal cancer patients with signet-ring cell carcinoma (SRCC) are still not clear. Materials and Methods: A retrospective comparison was made of the data of signet-ring cell colorectal carcinoma patients operated on between 2009 and 2018 in respect of clinicopathological and operative results, morbidity, mortality, and long-term survival. Results: The study included a total of 34 patients comprising 26 (76%) males and 8 (24%) females with a mean age of 58 ± 11.7 years. Incidence of SRCC was determined as 1.8%. Lymphovascular invasion was determined in 22 (64%) patients. Tumors were determined as stage T2 in 8 (32%) patients, stage T3 in 9 (36%), and stage T4 in 8 (32%). According to the TNM classification, 5 (14.7%) patients were diagnosed with stage 1, 7 (20.6%) with stage 2, 15 (44.1%) with stage 3, and 7 (20.6%) with stage 4. The mean follow-up period was 40.6 ± 30.4 months, and mean disease-free follow-up was determined as 33.1 ± 36.1 months. Fifteen (44.1%) patients died because of the disease. Conclusion: Although SRCC is a poor prognostic factor, it should be kept in mind when determining adjuvant therapies and prognosis of patients determined with advanced-stage SRCC.

Patients with non-muscle invasive bladder cancer (NMIBC) undergo lifelong monitoring based on repeated cystoscopy and urinary cytology due to the high recurrence rate of this tumor. Nevertheless, these techniques have some drawbacks, namely, low accuracy in detection of low-grade tumors, omission of pre-neoplastic lesions and carcinomas in situ (CIS), invasiveness, and high costs. This work aims to identify a urinary metabolomic signature of recurrence by proton Nuclear Magnetic Resonance (1H NMR) spectroscopy for the follow-up of NMIBC patients. To do this, changes in the urinary metabolome before and after transurethral resection (TUR) of tumors are analyzed and a Partial Least Square Discriminant Analysis (PLS-DA) model is developed. The usefulness of this discriminant model for the detection of tumor recurrences is assessed using a cohort of patients undergoing monitoring. The trajectories of the metabolomic profile in the follow-up period provide a negative predictive value of 92.7% in the sample classification. Pathway analyses show taurine, alanine, aspartate, glutamate, and phenylalanine perturbed metabolism associated with NMIBC. These results highlight the potential of 1H NMR metabolomics to detect bladder cancer (BC) recurrences through a non-invasive approach.