Alkaline-phosphatase-based nanostructure assemblies for electrochemical detection of microRNAs

Different nanoarchitectures, rich in enzyme labels, are herein investigated for signal amplification in the electrochemical detection of nucleic acids and in particular of microRNAs. Dendritic amplification, accomplished by the use of streptavidin and biotinylated alkaline phosphatase, and enzymedecorated liposomes are used as labels to amplify the microRNA-sensing, by their association to the probe-microRNA hybrid generated onto a gold transducer. Differential pulse voltammetry and faradaic impedance spectroscopy were employed to characterize these different amplification routes

custom splint application

To evaluate the clinical effectiveness of wrist splint usage arranged by determining the optimal position on which the median nerve is compressed the least through sonographic examination for patients with carpal tunnel syndrome (CTS). This study was a prospective, clinical trial with a 6-week follow-up. Twenty-four patients diagnosed clinically and electromyographically with CTS were included in the study. A total of 37 wrists were studied on. When the patients were grouped according to the optimal position, Group I comprising 16 (43.24%) wrists was at 15 degree flexion, Group II comprising 12 (32.43%) wrists was neutral, Group III comprising 6 (16.22%) wrists was at 15A degrees extension and Group IV comprising 3 (8.11%) wrists was at 30A degrees extension configurations. Groups I, II and III were included in clinical follow-up. Symptom severity score (SSS), functional status score (FSS), Grip strength and Pinch strength were used for the clinical follow-up and evaluation of the patients. When pre- and post-treatment were compared, a statistically significant recovery was detected in all three groups in respect to SSS (Group I P 0.05). In this study, by sonographic examination of the patients included in the study with CTS, we saw that the optimal position on which the median nerve is compressed the least varies depending on the individual and we determined that this position was 15A degrees flexion most frequently for our patients. We also observed that in clinical follow-up of wrist splint usage arranged on custom optimal position results in significant recovery

Candida albicans V132 induces trained immunity and enhances the responses triggered by the polybacterial vaccine MV140 for genitourinary tract infections

Introduction: Recurrent urinary tract infections (RUTIs) and recurrent vulvovaginal candidiasis (RVVCs) represent major healthcare problems all over the world. Antibiotics and antifungals are widely used for such infectious diseases, which is linked with microbial resistances and microbiota deleterious effects. The development of novel approaches for genitourinary tract infections (GUTIs) such as trained immunity-based vaccines (TIbV) is therefore highly required. MV140 is a sublingual whole-cell heat-inactivated polybacterial preparation with demonstrated clinical efficacy for RUTIs. The sublingual heat-inactivated Candida albicans vaccine V132 has been developed for RVVCs. We previously showed that the combination of MV140 and V132 promotes potent Th1/Th17 and regulatory T-cell responses against antigens contained in the formulation and unrelated antigens. The specific contribution of each preparation to such effects and the underlying molecular mechanisms remain incompletely understood. Methods: PBMC and monocytes were isolated from healthy donors and in vitro stimulated with V132, MV140 or MV140/V132. After 6 days of resting, cells were reestimulated with LPS and MV140. Analysis of cytokine production by ELISA, Seahorse assays for functional metabolic experiments and chromatin immunoprecipitation assays were performed. BALB/c mice were intraperitoneally and sublingually immunized with V132. Results: We uncover that V132 induces trained immunity in human PBMCs and purified monocytes, significantly increasing the responses triggered by purified monocytes, significantly increasing the responses triggered by subsequent stimulation with MV140. Mechanistically, V132 drives metabolic rewiring towards increased glycolysis and oxidative phosphorylation and induces epigenetic reprogramming that enhances the transcription of the pro-inflammatory genes IL6 and TNFA. Splenocytes and peritoneal cells from V132-immunize mice show increased responses upon in vitro stimulation with MV140. Remarkably, splenocytes from sublingually V132-immunized and MV140 in vivo treatment mice show stronger Th17 responses than mice exposed to excipients upon in vitro stimulation with MV140. Conclusion: Overall, we provide novel mechanistic insights into how V132- induced trained immunity enhances both innate and adaptive immune responses triggered by MV140, which might open the door for new interventions for GUTIs with important clinical implications.MINECOThe Netherlands Organization for Scientific ResearchNetherlands Organization for Scientific ResearchERC Advanced GrantDepto. de Bioquímica y Biología MolecularFac. de Ciencias QuímicasTRUEpu

Dimethyl itaconate induces long-term innate immune responses and confers protection against infection

Itaconate is an immunomodulatory metabolite produced by immune cells under microbial stimulation and certain pro-inflammatory conditions and triggers antioxidant and anti-inflammatory responses. We show that dimethyl itaconate, a derivative of itaconate previously linked to suppression of inflammation and widely employed as an alternative to the endogenous metabolite, can induce long-term transcriptional, epigenomic, and metabolic changes, characteristic of trained immunity. Dimethyl itaconate alters glycolytic and mitochondrial energetic metabolism, ultimately leading to increased responsiveness to microbial ligand stimulation. Subsequently, mice treated with dimethyl itaconate present increased survival to infection with Staphylococcus aureus. Additionally, itaconate levels in human plasma correlate with enhanced ex vivo pro-inflammatory cytokine production. Collectively, these findings demonstrate that dimethyl itaconate displays short-term anti-inflammatory characteristics and the capacity to induce long-term trained immunity. This pro-and anti-inflammatory dichotomy of dimethyl itaconate is likely to induce complex immune responses and should be contemplated when considering itaconate derivatives in a therapeutic context. Proteomic