Elaboración de kits de diagnóstico clínico mediante metodología de ELISA en diferentes enfermedades metabólicas

Los errores congénitos del metabolismo (ECM) son un grupo de enfermedades hereditarias causadas por defectos genéticos en las rutas metabólicas. Aunque existen más de 7000, la frecuencia de cada de una de ellas es tan baja que se consideran enfermedades raras y, actualmente, sólo la mitad son diagnosticables. El elevado coste de investigación ralentiza el desarrollo de métodos de diagnóstico más eficientes y posibles tratamientos. Una de los ECM más frecuente en España es la enfermedad de Gaucher, en la cual una mutación en el gen GBA lleva a la deficiencia en la actividad de la enzima β‑glucocerebrosidasa, provocando la acumulación de glucosilceramida en los lisosomas de macrófagos. La forma más común de Gaucher (94%) es el tipo 1, en el cual se aprecian manifestaciones óseas en más del 90% de los casos. En los últimos años se ha observado que estos pacientes presentan niveles elevados de catepsina K en suero. La catepsina K es una proteína esencial en la reabsorción ósea en personas normales. Es secretada por los osteoclastos hacia las zonas de contacto entre estas células y la superficie del hueso, donde participa en la degradación de la matriz orgánica mediante la escisión de las proteínas colágeno 1 y osteonectina. Los niveles elevados de catepsina K en pacientes de Gaucher podrían ser un indicativo del mayor nivel de degradación ósea que presentan estos pacientes y, por tanto, podrían utilizarse para el diagnóstico y seguimiento de la enfermedad. Se pretende desarrollar un kit para el diagnóstico y el seguimiento de la enfermedad de Gaucher tipo I basado en la metodología ELISA. Para ello, se va a poner a punto un procedimiento de ELISA Sándwich Heterólogo para la detección de catepsina K en muestras de suero de pacientes. Los resultados se compararán con niveles de quitotriosidasa de estos pacientes, el marcador utilizado actualmente para el diagnóstico de la enfermedad. Además, se contrastará con un análisis de otros marcadores de formación y destrucción ósea en los mismos pacientes. Los primeros resultados obtenidos muestran que existe una correlación clara entre los niveles de catepsina K en suero y la gravedad de la enfermedad en cada paciente, determinada por los niveles de quitotriosidasa. El análisis de los otros marcadores de formación y degradación ósea arroja resultados positivos. Es necesario llevar a cabo una validación completa del kit ELISA que se está desarrollando, así como comparar los resultados con kits existentes en el mercado

Determination of microbiological and sensory parameters of fish fillets with propolis preserved under refrigeration

ABSTRACT Objective: To determine the ability of propolis preservative cachama fillets during refrigerated storage. Materials and method. Ethanol extracts of propolis (EEP) were used in cachama fillets. The treatments included: i) 96% ethanol alcohol as the control; ii) 0.8% EEP; iii) 1.2% EEP; and iv) liquid smoke. A in vitro analysis was used to determine the inhibitory effect of propolis on Staphylococcus aureus, Escherichia coli, Salmonella sp. and Clostridium sp. and on the fish matrix to determine the mesophiles, psychrotrophiles, total coliforms, fecal coliforms, sulphite reducing spores and the presence of Salmonella. Results. The results of the in vitro analysis demonstrated the control that the EEP had over the evaluated microorganisms without presenting significant differences between the different concentrations (p>0.05). The analyses of the fish fillet matrix presented acceptable contents for the evaluated microorganisms in the treatments with EEP. A different situation was seen in the treatment with liquid smoke and the control, which had samples that where rejected after 20 days of storage. The sensory analysis showed acceptance for the samples with EEP until the end of the storage period but low marks for the treatment with liquid smoke and the control. Conclusions. The EEP used in this study could be effective for the control of Gram positive bacteria and some Gram negative bacteria that are present in cachama fillets; and could be an alternative to the use of chemical preservatives

Topological effects in ultrafast photoinduced processes between flurbiprofen and tryptophan in linked dyads and within human serum albumin

[EN] The interaction dynamics between flurbiprofen (FBP) and tryptophan (Trp) has been studied in covalently linked dyads and within human serum albumin (HSA) by means of fluorescence and ultrafast transient absorption spectroscopy. The dyads have proven to be excellent models to investigate photoinduced processes such as energy and/or electron transfer that may occur in proteins and other biological media. Since the relative spatial arrangement of the interacting units may affect the yield and kinetics of the photoinduced processes, two spacers consisting of amino and carboxylic groups separated by a cyclic or a long linear hydrocarbon chain (1 and 2, respectively) have been used to link the (S)- or (R)-FBP with the (S)-Trp moieties. The main feature observed in the dyads was a strong intramolecular quenching of the fluorescence, which was more important for the (S,S)- than for the (R,S)- diastereomer in dyads 1, whereas the reverse was true for dyads 2. This was consistent with the results obtained by simple molecular modelling (PM3). The observed stereodifferentiation in (S,S)-1 and (R,S)-1 arises from the deactivation of (1)Trp*, while in (S,S)-2 and (R,S)-2 it is associated with (FBP)-F-1*. The mechanistic nature of (FBP)-F-1* quenching is ascribed to energy transfer, while for (1)Trp* it is attributed to electron transfer and/or exciplex formation. These results are consistent with those obtained by ultrafast transient absorption spectroscopy, where (FBP)-F-1* was detected as a band with a maximum at ca. 425 nm and a shoulder at similar to 375 nm, whereas Trp did not give rise to any noticeable transient. Interestingly, similar photoprocesses were observed in the dyads and in the supramolecular FBP@HSA complexes. Overall, these results may aid to gain a deeper understanding of the photoinduced processes occurring in protein-bound drugs, which may shed light on the mechanistic pathways involved in photobiological damage.Grant PID2020-115010RB-I00 funded by MCIN/AEI/10.13039/501100011033 and grant from Conselleria d'Innovacio, Universitats, Ciencia i Societat Digital (CIAICO/2021/061) are gratefully acknowledged.Tamarit-Mayo, L.; García-Gabarda, L.; Jiménez, MC.; Miranda, MA.; Vayá Pérez, I. (2023). Topological effects in ultrafast photoinduced processes between flurbiprofen and tryptophan in linked dyads and within human serum albumin. Physical Chemistry Chemical Physics. 25(23):16148-16156. https://doi.org/10.1039/d3cp01082a1614816156252

Informe de seguimiento de la actividad de los Ministerios en relación con los Programas del Marco General para la Mejora de la Calidad de la Administración General del Estado durante 2011: Real Decreto 951/2005, de 29 de julio, por el que se establece el Marco General para la Mejora de la Calidad en la AGE

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The upper Valanginian of the Oliva section (Prebetic Zone, Valencia): facies analysis, biostratigraphy, C-isotope stratigraphy and organic geochemistry

The Upper Valanginian stratigraphic section of the sierra de la Oliva (Prebetic, Valencia province) records a significant sedimentary episode which has been named as ”Weissert Event”.This event is characterized by a positive excursion in the δ13C profile, which have been considered to be linked to a global perturbation in the carbon cycle, with environmental consequences that have been the object of debate, especially the degree of oxygenation of the sea bottom waters.We studied the sedimentary evolution during the late Valanginian in a shallow platform setting, beginning with the drowning of the early Valanginian carbonate platform, followed by a succession of shallowing- upwards sequences, which define a general deepening-upward evolution. The δ13C profile records a positive excursion during the late Valanginian, and the biomarker study has revealed the episodic development of anoxia, coeval to the deposition of organic rich facies, occurred during the episode of maximum deepening of the platform.This study has demonstrated that the “Weissert Event” has been recorded in the Prebetic platform, and that the environmental perturbations gave place to the occasional development of anoxia in a context of high organic productivityLa sección estratigráfica del Valanginiense superior de la sierra de la Oliva (Prebético, provincia de Valencia) registra un interesante episodio sedimentario que ha sido denominado “Evento Weissert”. Este evento está caracterizado por una excursión positiva en los valores del δ13C, que se considera ligada a una perturbación global en el ciclo del carbono, cuyas consecuencias ambientales son objeto de discusión, especialmente las condiciones de oxigenación en los fondos marinos. El estudio presentado aquí muestra la evolución sedimentaria del Valanginiense superior en un ámbito de plataforma somera, que comienza con el drowning de la plataforma carbonatada del Valanginiense inferior, y continúa con una sucesión de secuencias de somerización, que en conjunto definen una evolución general de profundización. El perfil de δ13C registra una excursión positiva durante el Valanginiense superior, y el estudio de los biomarcadores ha puesto de manifiesto el desarrollo puntual de anoxia, simultáneo al depósito de facies ricas en materia orgánica, ocurrido en el momento de mayor profundización de la plataforma. Este estudio demuestra que el denominado “EventoWeissert” quedó reflejado en la plataforma Prebética, y que las perturbaciones ambientales locales dieron lugar al desarrollo puntual de anoxia en un contexto de elevada productividad orgánic

Evaluation of a Multiplex ELISA for Autoantibody Profiling in Patients with Autoimmune Connective Tissue Diseases

The performance of immunoassays for the detection of autoantibodies is of critical importance in the diagnosis and assessment of patients with autoimmune connective tissue diseases (ACTD). Our objective was to compare the features of two multiplexed assays—INNO-LIA ANA and Gennova-PictArray ENA ELISA—for measurement of multiple autoantibodies and their utility as a clinical tool in ACTD diagnosis. The antigens included SS-A/Ro (60 and 52), SSB/La, Sm, Sm/RNP, CENP-B, Jo-1, and Scl-70. Stored sera from 85 ACTD patients and 80 controls consisting of patients with vasculitis, rheumatoid arthritis and infectious diseases, as well as healthy subjects were analyzed jointly with clinical and laboratory data. Agreement between the two methods varied between 58 and 99% (Cohen’s kappa: 0.21–0.71) mostly for SSA and SSB. The frequency of specific autoantibodies measured using the two methods was more variable for SSA, SSB, and RNP/Sm. There were a higher number of ambiguous results when using INNO-LIA. The optimized cut-off values of the Gennova-PictArray resulted in over 99% specificities in samples obtained from the control group. Sensitivity patterns were more accurate in Gennova-PictArray than in INNO-LIA, as suggested in previously reported studies. A third method could be applied to determine which of the two methods is more accurate

Novel Fluorescent Labeled Octasilsesquioxanes Nanohybrids as Potential Materials for Latent Fingerprinting Detection

The recent demand for fluorescent labeled materials (FLMs) in forensic security concepts such as latent fingerprints (LFs) that encodes information for anti-counterfeiting and encryption of confidential data makes necessary the development of building new and innovative materials. Here, novel FLMs based on Polyhedral Oligomeric Silsesquioxanes (POSS) functionalized with fluorophores via “click” reactions have been successfully synthesized and fully characterized. A comprehensive study of their photophysical properties has displayed large Stokes’s shift together with good photostability in all cases, fulfilling the fundamental requisites for any legible LF detection on various surfaces. The excellent performance of the hetero-bifunctional FLM in the visualization of LF is emphasized by their legibility, selectivity, sensitivity and temporal photostability. In this study, development mechanisms have been proposed and the overall concept constitute a novel approach for vis-à-vis forensic investigations to trace an individual’s identity.Alexander von Humboldt FoundationGeneralitat ValencianaUniversität RegensburgUniversidad de La LagunaMinisterio de Ciencia, Innovación y Universidade

Transient absorption spectroscopy detection of sensitized delayed fluorescence in chiral benzophenone/naphthalene systems

Transient absorption spectroscopy has proven to be a powerful tool to investigate the formation and decay of excited singlet states upon triplet–triplet annihilation, following T–T energy transfer from a selectively excited sensitizer. Thus, upon selective excitation of benzophenone (BZP) by laser flash photolysis (LFP) at λ = 355 nm in the presence of naphthalene (NPT), a negative band centered at 340 nm has been detected, with growth and decay in the microsecond timescale. It has been assigned to the P-type NPT delayed-fluorescence. In the case of chiral BZP/NPT systems, stereodifferentiation has been observed in the kinetics of the involved photophysical processesFinancial support from the MICINN (Grant CTQ2010-14882 and predoctoral fellowship to P. B.) is gratefully acknowledged.Bonancía Roca, P.; Jiménez Molero, MC.; Miranda Alonso, MÁ. (2011). Transient absorption spectroscopy detection of sensitized delayed fluorescence in chiral benzophenone/naphthalene systems. Chemical Physics Letters. 515(1-3):194-196. https://doi.org/10.1016/j.cplett.2011.08.096S1941965151-

Recent applications of biphotonic processes in organic synthesis

Currently, evolution of chemical transformations by visible light irradiation is highly desirable from cost, safety, availability, and environmental friendliness points of view. Besides, activation of less reactive substrates under very mild conditions becomes one of the most challenging tasks in organic synthesis. However, the insufficient energy provided by one photon of visible light for their activation definitely makes necessary the development of new protocols together with the design of new photocatalytic systems to overcome this limitation. In this context, the implementation of biphotonic processes has been found to be a solution for these drawbacks. This new mechanistic paradigm which combines light upconversion processes with energy/electron transfers holds great potential for high energy demanding bond activations, expanding the accessible reactivity window. Here, we wish to highlight the recent applications of biphotonic processes in organic synthesis.Generalitat ValencianaGobierno de Españ