New Advances in the Exploration of Esterases with PET and Fluorescent Probes.

Esterases are hydrolases that catalyze the hydrolysis of esters into the corresponding acids and alcohols. The development of fluorescent probes for detecting esterases is of great importance due to their wide spectrum of biological and industrial applications. These probes can provide a rapid and sensitive method for detecting the presence and activity of esterases in various samples, including biological fluids, food products, and environmental samples. Fluorescent probes can also be used for monitoring the effects of drugs and environmental toxins on esterase activity, as well as to study the functions and mechanisms of these enzymes in several biological systems. Additionally, fluorescent probes can be designed to selectively target specific types of esterases, such as those found in pathogenic bacteria or cancer cells. In this review, we summarize the recent fluorescent probes described for the visualization of cell viability and some applications for in vivo imaging. On the other hand, positron emission tomography (PET) is a nuclear-based molecular imaging modality of great value for studying the activity of enzymes in vivo. We provide some examples of PET probes for imaging acetylcholinesterases and butyrylcholinesterases in the brain, which are valuable tools for diagnosing dementia and monitoring the effects of anticholinergic drugs on the central nervous system.

Human blastocysts uptake extracellular vesicles secreted by endometrial cells containing miRNAs related to implantation.

STUDY QUESTION: Are the extracellular vesicles (EVs) secreted by the maternal endometrium uptaken by human embryos and is their miRNA cargo involved in implantation and embryo development? SUMMARY ANSWER: Data suggest that EVs secreted by human endometrial epithelial cells are internalized by human blastocysts, and transport miRNAs to modulate biological processes related to implantation events and early embryo development. WHAT IS KNOWN ALREADY: Successful implantation is dependent on coordination between maternal endometrium and embryo, and EVs role in the required cell-to-cell crosstalk has recently been established. In this regard, our group previously showed that protein cargo of EVs secreted by primary human endometrial epithelial cells (pHEECs) is implicated in biological processes related to endometrial receptivity, embryo implantation, and early embryo development. However, little is known about the regulation of these biological processes through EVs secreted by the endometrium at a transcriptomic level. STUDY DESIGN, SIZE, DURATION: A prospective descriptive study was performed. Endometrial biopsies were collected from healthy oocyte donors with confirmed fertility on the day of oocyte retrieval, 36 h after the LH surge. pHEECs were isolated from endometrial biopsies (n = 8 in each pool) and cultured in vitro. Subsequently, conditioned medium was collected and EVs were isolated and characterized. Uptake of EVs by human blastocysts and miRNA cargo of these EVs (n = 3 pools) was analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: EVs were isolated from the conditioned culture media using ultracentrifugation, and characterization was performed using western blotting, nanoparticle tracking analysis, and transmission electron microscopy. EVs were fluorescently labeled with Bodipy-TR ceramide, and their uptake by human blastocysts was analyzed using confocal microscopy. Analysis of the miRNA cargo of EVs was performed using miRNA sequencing, target genes of the most expressed miRNA were annotated, and functional enrichment analysis was performed. MAIN RESULTS AND THE ROLE OF CHANCE: EVs measured 100-300 nm in diameter, a concentration of 1.78 × 1011 ± 4.12 × 1010 (SD) particles/ml and expressed intraluminal protein markers Heat shock protein 70 (HSP70) and Tumor Susceptibility Gene 101 (TSG101), in addition to CD9 and CD81 transmembrane proteins. Human blastocysts efficiently internalized fluorescent EVs within 1-2 h, and more pronounced internalization was observed in the hatched pole of the embryos. miRNA-seq analysis featured 149 annotated miRNAs, of which 37 were deemed most relevant. The latter had 6592 reported gene targets, that in turn, have functional implications in several processes related to embryo development, oxygen metabolism, cell cycle, cell differentiation, apoptosis, metabolism, cellular organization, and gene expression. Among the relevant miRNAs contained in these EVs, we highlight hsa-miR-92a-3p, hsa-let-7b-5p, hsa-miR-30a-5p, hsa-miR-24-3p, hsa-miR-21-5p, and hsa-let-7a-5p as master regulators of the biological processes. LIMITATIONS, REASONS FOR CAUTION: This is an in vitro study in which conditions of endometrial cell culture could not mimic the intrauterine environment. WIDER IMPLICATIONS OF THE FINDINGS: This study defines potential biomarkers of endometrial receptivity and embryo competence that could be useful diagnostic and therapeutic targets for implantation success, as well as open insight further investigations to elucidate the molecular mechanisms implicated in a successful implantation. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by the Spanish Ministry of Education through FPU awarded to M.S.-B. (FPU18/03735), the Health Institute Carlos III awarded to E.J.-B. (FI19/00110) and awarded to H.F. by the Miguel Servet Program 'Fondo Social Europeo «El FSE invierte en tu futuro¼' (CP20/00120), and Generalitat Valenciana through VALi+d Programme awarded to M.C.C.-G. (ACIF/2019/139). The authors have no conflicts of interest to disclose. TRIAL REGISTRATION NUMBER: N/A.

Extracellular vesicles secreted by adenomyosis endometrial organoids contain miRNAs involved in embryo implantation and pregnancy.

RESEARCH QUESTION: Do extracellular vesicles secreted by the endometrium of women with adenomyosis contain miRNAs involved in adenomyosis-related infertility? DESIGN: A descriptive study using organoids from eutopic endometrium of women with adenomyosis (n = 4) generated and differentiated to secretory and gestational phases, in which miRNA cargo from extracellular vesicles secreted by these differentiated organoids in each phase was analysed by next-generation sequencing. miRNAs in secretory-extracellular vesicles and gestational-extracellular vesicles were selected based on the counts per million. miRNAs target genes in each phase were obtained from miRNet and gene ontology was used for enrichment analysis. RESULTS: miRNA sequencing identified 80 miRNAs in secretory-phase extracellular vesicles, including hsa-miR-21-5p, hsa-miR-24-3p, hsa-miR-26a-5p, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-miR-200c-3p and hsa-miR-423a-5p, related to adenomyosis pathogenesis and implantation failure. Further, 60 miRNAs were identified in gestational-phase extracellular vesicles, including hsa-miR-21-5p, hsa-miR-26a-5p, hsa-miR-30a-5p, hsa-miR-30c-5p, hsa-miR-222-3p and hsa-miR-423a-5p were associated with preeclampsia and miscarriage. Among the target genes of these miRNAs, PTEN, MDM4, PLAGL2 and CELF1, whose downregulation (P = 0.0003, P < 0.0001, P = 0.0002 and P = 0.0003, respectively) contributes to adenomyosis pathogenesis, and impaired early embryo development, leading to implantation failure and miscarriage, are highlihghted. Further, functional enrichment analyses of the target genes revealed their involvement in cell differentiation, proliferation, apoptosis, cell cycle regulation and response to extracellular stimuli. CONCLUSIONS: Eutopic endometrium in secretory and gestational phase from women with adenomyosis releases extracellular vesicles containing miRNAs involved in adenomyosis progression, impaired embryo implantation and pregnancy complications.

Breakthrough Infections Following mRNA SARS-CoV-2 Vaccination in Kidney Transplant Recipients.

BACKGROUND: The clinical effectiveness of coronavirus disease 2019 (COVID-19) vaccination in kidney transplant (KT) recipients is lower than in the general population. METHODS: From April to October 2021, 481 KT recipients with COVID-19, included in the Spanish Society of Nephrology COVID-19 Registry, were analyzed. Data regarding vaccination status and vaccine type were collected, and outcomes of unvaccinated or partially vaccinated patients (n = 130) were compared with fully vaccinated patients (n = 351). RESULTS: Clinical picture was similar and survival analysis showed no differences between groups: 21.7% of fully vaccinated patients and 20.8% of unvaccinated or partially vaccinated died (P = 0.776). In multivariable analysis, age and pneumonia were independent risk factors for death, whereas vaccination status was not related to mortality. These results remained similar when we excluded patients with partial vaccination, as well as when we analyzed exclusively hospitalized patients. Patients vaccinated with mRNA-1273 (n = 213) showed a significantly lower mortality than those who received the BNT162b2 vaccine (n = 121) (hazard ratio: 0.52; 95% confidence interval, 0.31-0.85; P = 0.010). CONCLUSIONS: COVID-19 severity in KT patients has remained high and has not improved despite receiving 2 doses of the mRNA vaccine. The mRNA-1273 vaccine shows higher clinical effectiveness than BNT162b2 in KT recipients with breakthrough infections. Confirmation of these data will require further research taking into account the new variants and the administration of successive vaccine doses.

Madurez de las vellosidades coriales en relación con malformaciones del tubo neural durante el tercer trimestre del embarazo / Maturity of corial villi in relation with neural tube malformations during the third trimester of pregnancy

Evaluar las características que determinan el grado de desarrollo de la vellosidad placentaria en los casos de las malformaciones del tubo neural. Laboratorio de microscopia electrónica del CIADANA, Maracay. Se utilizó un protocolo de observación, usando la microspía de luz, basado en los tipos de vellosidades y de membranas vásculosincitiales; incremento de citotrofoblastos, calcificación y puente sincitiales en los casos de acraneo-excencefalia; cráneo-raquisquisis con trisomía 18, mielomeningocele con hidrocefalia asociada con otras malformaciones congénitas, mielomeningocele lumbar, anencefalia y malformación de Chiari tipo II apareando con controles a la misma edad gestacional sin malformaciones. Las cifras en porcentajes de vellosidades maduras e inmaduras y de membranas vásculosincitiales periféricas o centrales exhiben variaciones según la malformación con predominio de inmaduras. Se encontró daño hipóxico velloso extenso. Estos resultados revelaron inmadures vellosa persistente correspondiéndose con la mala perfusión placentaria y disfunción de la capacidad de transferencia. Estos hallazgos son de interés cuando otros test como la determinación de alfafetoproteínas y estudios ultrasonográficos son empleados al brindar información adicional útil en el diagnóstico complementario

Observaciones en la hiperplasia mesenquimal placentaria con el microscopio electrónico de barrido / Observations in the placental mesenchymal hyperplasia by scanning electron microscopy

Material de una placenta de embarazo con diagnóstico de enfermedad mesenquimatosa placentaria a las 27 semanas de gestación y dolicocefalia fetal fue obtenido para ser procesado con las técnicas de microscopia electrónica de barrido previo análisis macro y microscópico del mismo. El examen macroscopico mostró placentomegalia, vellosidades troncales hidrópticas de tamaño entre 2 y 35 mm, placa carionica comprimida, hematoma retroplacentario

La vellosidad placentaria en caso de anemia drepanocítica / The placentary villi to case of drepanocytic anemia

Describir los efectos de la hemoglobinopatía SS en la organización histoarquitectónica de las últimas ramificaciones periféricas del árbol velloso maduro. Se tomaron imágenes de micrografías de luz y electrónica de barrido con las alteraciones sufridas. Centro de Investigación y Análisis Docente Asistencial del Núcleo Aragua. facultad de Ciencias de la Salud. Laboratorio de Microscopia Electrónica. Maracay. Estado Aragua. Deposición de fibrinoide intravelloso incrementado se observó en las diferentes vellosidades infiltrado por células X trofoblástica. Es notorio el incremento de nódulos sincitiales. Hay necrosis del trofoblasto. El estroma contiene vasos dilatados y congestionados con uno o tres eritrocitos nucleados. Se identificó hemorragia estronal, necrosis endotelial, fibrosis y desorganización estromal por edema hidrópico, calcificación, infartos vellosidades intermedias maduras con deficiencia de vellosidad terminal. Los resultados indican cambios degenerativos a nivel de la barrera placentaria que pronostican retardo del crecimiento intrauterino similar al encontrado con mala perfusión de la vellosidad placentaria de origen fetal o materno.

Vellosidades placentarias terminales filiformes en la anemia drepanocítica / Terminal filiform placental villi in sickle cell disease

Describir las vellosidades placentarias filiformes, descritas anteriormente como vellosidades intermedias maduras con deficiencia de vellosidades terminales, en la hemoglobinopatía ss, con técnicas de microscopia de luz y electrónica de barrido, es el propósito de nuestro trabajo. Cincuenta láminas de hemotoxilina-eosina fueron observadas de regiones marginales, centrales e intermedias próximas a la placa basal de la placenta y especímenes fueron tomados para su observación con el microscopio electrónico de barrido. Los resultados indican la presencia de vellosidades terminales filiformes, que sobresalen del conglomerado con un eje longitudinal muy alargado diferente a las vellosidades terminales normales. Se destacan por la ausencia de citotrofoblastos, trofoblasto sincisial de grosor variable y reducción de membrana vasculosincisiales. La distancia del centro de la luz del capilar a la superficie del trofoblasto es relativamente grande. Este último carácter y el estancamiento de eritrocitos en los capilares alargados dañan la superficie de intercambio de gases y nutrientes que pronostican retardo del crecimiento intrauterino

Vasculogénesis en la vellosidad placentaria humana de nueve semanas / Vaculogenesis in nine months human placental villi

Pertubaciones en el desarrollo vascular placentario demostrado por reducida ramificación capilar vellosa y alteraciones en la expresión de factores angiogénenicos han estado asociados con aborto, preeclapsia, retardo del crecimiento intrauterino y diabetes. Esta influencia negativa sobre el desarrollo fetal amerita el estudio urgente y detallado de la formación de vasos intravellositarios mediante técnicas de hematoxilina-eosina. Especímenes fueron tomados de un aborto terapéutico de nueve semanas de gestación exactamente fechada según fecha de última mestruación y datos ultrasonográficos. Los resultados revelan vellosidades intermedias inmaduras, troncales y numerosos macrófagos. Se observó el proceso de vasculogénesis desde la transformación de células mesenquimales a células hemangioblásticas productoras de células endoteliales, de los primitivos tubos capilares. Se observaron espacios claros entre dos o tres de estas últimas células que indican la formación de la futura luz del vaso. Cuatro tipos celulares parecen participar en la formación de vasos.

La membrana basal en caso de mola hidatidiforme completa / The basal menbrane in case of complete hydatidiform mole

Para la evaluación de la integridad de la menbrana basal así como sus características en caso de mola hidatidiforme completa como parte de la matriz extracelular subyacente al trofoblasto, fueron preparadas las láminas con las técnicas de ácido periódico reactivo de Schiff para polisacáridos complejos y nitrato de plata para proteínas. Las láminas positivas fueron seleccionadas para su observación con el microscopio electrónico de barrido. La membrana basal mostró engrosamiento, desprendimiento o separación de la membrana plasmática del sincitio, interrupciones o discontinuidades, adelgazamientos, bifurcaciones o reduplicaciones, zonas de mínima o exagerada hiperplasia pueden carecer de membrana basal que se altera en áreas de acentuado edema. Los cambios degenerativos en la membrana basal pudieran ser indicativos de la interacción de células trofoblásticas con la matrizextracelular, pronóstico de posible transformación maligna o del efecto provocado por un permanente transporte de fluido

La variabilidad morfológica de la vellosidad placentaria hidrópica / Morfological variability of the hydropic placentary villi

Este estudio se propone distinguir la diversidad de formas que adopta la vellosidad placentaria hidrópica en caso de aborto, mola hidatidiforme y displasia mesenquimal placentaria mediante una secuencia de imágenes logradas con microscopia de luz y electrónica de "barrido", Vesículas vellositarias procedentes de legrado, de diversos tamaños, fueron tomadas para ser procesadas con las técnicas rutinarias de hematoxilina-eosina y microscopia electrónica de "barrido". Los resultados en imágenes micrográficas demostraron la diferenciación histopatológica de la vellosidad en las tres lesiones mencionadas basadas en criterios morfológicamente ya establecidos en la litertura. Como hay diferencias en el origen, la morfología y la historia natural de estas vellosidades, esta secuencia nos permite identificar especímenes raros que se desvíen de las aquí mostradas