Enzymatic upgrading of nanochitin using an ancient lytic polysaccharide monooxygenase

Numerous enzymes have the potential to upgrade biomass, converting it into high-tech materials for new applications. However, the features of natural enzymes often limit their use beyond chemical conversion of the substrate. The development of strategies for the enzymatic conversion of biomass into high-value materials may broaden the range of applications of enzymes and enzyme design techniques. A relevant case is lytic polysaccharide monooxygenase (LPMO), a class of enzymes that catalyzes the oxidative cleavage of glycosidic bonds. Here, we show that an ancestral LPMO can generate chitin nanocrystals. Physicochemical characterization of the chitin nanocrystals demonstrates modifications that make it superior compared to chitin obtained by chemical treatments. We show that the nanocrystals are suitable for controlled 2D and 3D cell cultures, as well as for engineering a biomatrix that combines with graphene oxide, forming a hybrid conductive bioink.This work has been supported by grants PID2019-109087RB-I00 to R.P.-J. from Spanish Ministry of Science and Innovation. This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 964764 to R.P.-J. ‘Materials + Technologies’ Research Group also acknowledges UPV/EHU and the Basque Government in the frame of “Research Group” (GIU 18/216) and “Grupos Consolidados” (IT776-13), respectively. We also thank Gipuzkoako Foru Aldundia for financial Support. HEK293T cells were a kind gift from Dr. Maria Muñoz Caffarel (Biodonostia, San Sebastian, Spain)

Aproximación multidisciplinar a la supervisión del practicum en las carreras de educación, medicina, odontología, informática, comunicación y documentación

Memoria ID-035. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2013-2014

Estudio de la estabilidad y evaluación de la toxicidad de las mezclas estandarizadas de terapia triple intratecal con metotrexato citarabina e hidrocortisona

Objetivos: Los objetivos principales del estudio fueron: 1) Estudiar la estabilidad fisicoquímica, en condiciones de uso, de cuatro mezclas estandarizadas triple intratecales (TIT) de metotrexato, citarabina e hidrocortisona conservadas a temperatura ambiente y en refrigeración. 2) Evaluar la toxicidad de la administración de tratamiento triple intratecal estandarizado como profilaxis y tratamiento de la enfermedad leptomeníngea en pacientes adultos y pediátricos. Metodología: La estabilidad química de la muestra se midió por triplicado mediante cromatografía líquida de alta eficacia a las 0, 3, 6, 12 y 24 horas y 2, 4, 5, 6 y 7 días de las mezclas conservadas a temperatura ambiente y bajo refrigeración, se determinó también a esos tiempo el pH y la osmolaridad de las mezclas. La concentración de fármacos a tiempo 0 se definió como el 100% y se consideró que los fármacos eran estables químicamente si la concentración era igual o mayor al 90% de la inicial. Los valores de pH y osmolaridad aceptados como válidos fueron cercanos al rango fisiológico del líquido cefalorraquídeo, considerándose las mezclas triple intratecales estables si el rango de pH y osmolaridad era de 7–7,5 y de 280–310 mOsm/kg, respectivamente. La evaluación de la toxicidad se realizó de forma prospectiva en todos los pacientes adultos y pediátricos a los que se les administró quimioterapia TIT estandarizada para el tratamiento o la profilaxis de la enfermedad leptomeníngea en el Hospital Universitario Virgen de la Arrixaca desde el 1 de enero de 2013 hasta el 31 de Julio de 2014 (18 meses). Se realizó un seguimiento de todos los pacientes desde la administración de la primera TIT del estudio hasta el 31 de Julio de 2014, excepto exitus. Los eventos adversos fueron detectados por el médico responsable del paciente y registrados por el investigador principal. La definición y gradación de los eventos adversos se realizó según la escala CTCAE v.4.0. Conclusiones: 1) Las concentraciones de metotrexato sódico, citarabina e hidrocortisona fosfato sódico de las cuatro mezclas triple intratecales estandarizadas, conservadas a 25ºC y 2-8ºC no variaron más del 10% respecto a las concentraciones iniciales a ninguno de los tiempos de medida, considerándose estables químicamente durante, al menos, 7 días. La osmolaridad de las mezclas se mantuvo durante los 7 días de estudio, mientras que el pH se situó dentro de rango de estudio durante 5-7 días en las mezclas refrigeradas y 48-96 horas en las conservadas a temperatura ambiente, en función de la mezcla. 2) En los pacientes pediátricos, se ha observado toxicidad en el 56,1% de los pacientes y el 16,7% de las administraciones y en los pacientes adultos se ha observado toxicidad en el 70% de los pacientes y el 39,3% de las administraciones. La toxicidad mayoritaria en los pacientes pediátricos fue la emesis, seguida de cefalea y dolor lumbar y en los adultos la cefalea, seguido de la emesis y la aparición de vértigos. La mayoría de los eventos adversos detectados durante el periodo de estudio, en la población global, fueron de grado 1-2. La frecuencia de aparición de toxicidad grado 3 fue del 4,9% en pacientes pediátricos y 3,2% en adultos. No se detectaron reacciones adversas de grado 4-5. Objectives: The main objectives of the study were: 1) Study the physicochemical stability of four standard triple intrathecal (TIT) mixtures of methotrexate, cytarabine and hydrocortisone stored at room temperature and refrigerated. 2) Evaluate the toxicity of intrathecal administration of standard triple therapy for prophylaxis and treatment of leptomeningeal disease in adults and pediatric patients. Methodology: The chemical stability of the sample was measured in triplicate by high-performance liquid chromatography at 0, 3, 6, 12 and 24 hours and 2, 4, 5, 6 and 7 days preserving the samples at room temperature and under refrigeration, pH and osmolarity of the mixtures were also determined at such time. The drug concentration at time 0 was defined as 100% and the drugs were considered chemically stable if the concentration was equal or greater than 90% of the initial. The pH and osmolarity were accepted as valid close to the physiological range of cerebrospinal fluid, considered stable triple mixtures intrathecal if the range of pH and osmolality was 7-7,5 and 280-310 mOsm/ kg, respectively. The toxicity evaluation was performed prospectively in all adult and pediatric patients who were administered chemotherapy TIT standardized for the treatment or prophylaxis of leptomeningeal disease at the University Arrixaca Hospital from 1st of January 2013 to 31st of July 2014 (18 months). The monitoring of all patients was performed from the administration of the first TIT study until 31st of July 2014, except for death. Adverse events were detected by the attending physician and recorded by the principal investigator. The definition and grading of adverse events was performed according to CTCAE v.4.0 scale. Conclusions: 1) The concentrations of sodium methotrexate, cytarabine and hydrocortisone sodium phosphate of the four triple intrathecal standardized mixtures, preserved at 25 ° C and 2-8ºC did not vary more than 10% compared to the initial concentrations any of the measured times, considering chemically stable for at least 7 days. Osmolarity mixtures remained during the 7 day of the study, while the pH was within range study for 5-7 days in refrigerated mixtures and 48-96 hours in room temperature ones, depending on the mixture. 2) In pediatric patients, toxicity has been observed in 56,1% of patients and 16,7% of administrations while in adult patients toxicity has been observed in 70% of patients and 39,3 % of administrations. The major toxicity in pediatric patients was emesis, followed by headache and low back pain and in adults was headache, followed by the appearance of emesis and dizziness. Most adverse events detected during the study period, in the overall population, were grade 1-2. The frequency of grade 3 toxicity was 4.9% in pediatric patients and 3.2% in adults. No grade 4-5 adverse reactions were detected

3D printed alginate-cellulose nanofibers based patches for local curcumin administration

Alginate and nanocellulose are potential biomaterials to be employed as bioinks for three-dimensional (3D) printing. Alginate-cellulose nanofibers (A-CNF) formulations with CNF amounts up to 5 wt% were developed and rheologically characterized to evaluate their printability. Results showed that formulations with less than 3 wt% CNF did not present suitable characteristics to ensure shape fidelity after printing. Selected A-CNF bioinks were 3D printed and freeze-dried to obtain porous scaffolds. Morphological and mechanical analysis were performed, showing that CNF contributed to the reinforcement of the scaffolds and modulated their porosity. The applicability for drug delivery was evaluated by the addition of curcumin to printable A-CNF formulations. The curcumin loaded bioinks were successfully 3D printed in patches and the in vitro release tests showed that alginate and CNF played an important role in curcumin stabilization, whereas the CNF content and the disintegration of the scaffold were essential in the release kinetics.Financial support from the University of the Basque Country (UPV/EHU) (GIU18/216 Research Group), from the Basque Government in the frame of Elkartek KK-2020/00053 and PIBA2020-1-0041 and from Spanish Ministry of Science, Innovation and Universities and European Union (MICINN/EU/FEDER) in the frame of MAT2016-76294-R and PID2019-105090RB-I00 projects, are gratefully acknowledged. Moreover, we are grateful to the Macrobehavior-Mesostructure-Nanotechnology SGIker unit of the UPV/EHU. Raquel Olmos Juste wishes to acknowledge the Ministry of Economy, Industry and Competitiveness for her PhD grant

Treatment adherence in patients older than 65 years who suffer early readmissions

Objective: Analyze the frequency of therapeutic noncompliance in patients who suffer early readmissions, and identify the factors associated with it. Method: Observational, descriptive study of three months duration (March - May 2014). All patients older than 65 years who readmitted in the 3-30 days following the last hospital discharge were included. We excluded programmed re-admissions and readmissions in the Intensive Care Unit. The variables collected were: age, sex, medical service, major diagnostic category, polypharmacy, number of days since the last hospital discharge, presence of hypertension and/or diabetes. The therapeutic compliance and the difficulty in the administration of medication were evaluated by means of the Morisky-Green test and the Haynes-Sackett test respectively. A descriptive analysis of the variables was carried out and they were related to the therapeutic adherence. The variables with statistical significance were included in a multivariate logistic regression model. Results: Fifty seven percent of the patients presented lack of adherence to pharmacological treatment. Twenty three percent had difficulty administering the medication. Eighty six percent had comorbidities (hypertension and/or diabetes) and 79% had a caregiver. Eighty six percent of patients were polymedicated (≥ 5 drugs). There is a relationship between lack of adherence and difficulty in the administration of medications (p=0.021), polypharmacy (p=0.002), and the presence of diabetes mellitus (p=0.018). Conclusions: Polymedication, the presence of diabetes mellitus and the existence of difficulty in the administration of medication are evidenced as prognostic factors of the lack of adherence to treatment in patients older than 65 years

La estímulación sensorial para el desarrollo competencial del alumno de educación especial

Seleccionado en la convocatoria: Ayudas a la innovación e investigación educativa en centros docentes de niveles no universitarios, Gobierno de Aragón 2010-11Proyecto del colegio Pío XII cuyo fin es la estimulación sensorial de los alumnos de educación especial. Sus objetivos son: presentar estímulos sensoriales; verbalizar oralmente las vivencias; manipular la pizarra táctil digital; fomentar el espíritu emprendedor; conocer el entorno próximo y sus propiedades; y trabajar con mayor profundidad aspectos básicos. Se diseñan actividades que se desarrollan en el gimnasio y en sala de psicomotricidad del centro, que contribuyen a la consecución de los objetivos. Se llevan a cabo actividades en el medio natural: visita de parques y entorno natural de Isin y excursión al acuario de Zaragoza.Gobierno de Aragón. Departamento de Educación, Cultura y DeporteAragónDirección General de Política Educativa; Avda. Gómez Laguna, 25, planta 2; 50009 Zaragoza; Tel. +34976715416; Fax +34976715496ES

Role of PATJ in stroke prognosis by modulating endothelial to mesenchymal transition through the Hippo/Notch/PI3K axis

Through GWAS studies we identified PATJ associated with functional outcome after ischemic stroke (IS). The aim of this study was to determine PATJ role in brain endothelial cells (ECs) in the context of stroke outcome. PATJ expression analyses in patient's blood revealed that: (i) the risk allele of rs76221407 induces higher expression of PATJ, (ii) PATJ is downregulated 24 h after IS, and (iii) its expression is significantly lower in those patients with functional independence, measured at 3 months with the modified Rankin scale ((mRS) ≤2), compared to those patients with marked disability (mRS = 4-5). In mice brains, PATJ was also downregulated in the injured hemisphere at 48 h after ischemia. Oxygen-glucose deprivation and hypoxia-dependent of Hypoxia Inducible Factor-1α also caused PATJ depletion in ECs. To study the effects of PATJ downregulation, we generated PATJ -knockdown human microvascular ECs. Their transcriptomic profile evidenced a complex cell reprogramming involving Notch, TGF-ß, PI3K/Akt, and Hippo signaling that translates in morphological and functional changes compatible with endothelial to mesenchymal transition (EndMT). PATJ depletion caused loss of cell-cell adhesion, upregulation of metalloproteases, actin cytoskeleton remodeling, cytoplasmic accumulation of the signal transducer C-terminal transmembrane Mucin 1 (MUC1-C) and downregulation of Notch and Hippo signaling. The EndMT phenotype of PATJ-depleted cells was associated with the nuclear recruitment of MUC1-C, YAP/TAZ, β-catenin, and ZEB1. Our results suggest that PATJ downregulation 24 h after IS promotes EndMT, an initial step prior to secondary activation of a pro-angiogenic program. This effect is associated with functional independence suggesting that activation of EndMT shortly after stroke onset is beneficial for stroke recovery

Glioblastoma Treatment: State-of-the-Art and Future Perspectives

(1) Background: Glioblastoma is the most frequent and lethal primary tumor of the central nervous system. Through many years, research has brought various advances in glioblastoma treatment. At this time, glioblastoma management is based on maximal safe surgical resection, radiotherapy, and chemotherapy with temozolomide. Recently, bevacizumab has been added to the treatment arsenal for the recurrent scenario. Nevertheless, patients with glioblastoma still have a poor prognosis. Therefore, many efforts are being made in different clinical research areas to find a new alternative to improve overall survival, free-progression survival, and life quality in glioblastoma patients. (2) Methods: Our objective is to recap the actual state-of-the-art in glioblastoma treatment, resume the actual research and future perspectives on immunotherapy, as well as the new synthetic molecules and natural compounds that represent potential future therapies at preclinical stages. (3) Conclusions: Despite the great efforts in therapeutic research, glioblastoma management has suffered minimal changes, and the prognosis remains poor. Combined therapeutic strategies and delivery methods, including immunotherapy, synthetic molecules, natural compounds, and glioblastoma stem cell inhibition, may potentiate the standard of care therapy and represent the next step in glioblastoma management research