Objective The aim of the study was to provide data on the fracture strength and fracture mode of monolithic high translucent Y-TZP crowns and porcelainveneered high translucent Y-TZP crown cores and to compare that data with the fracture strength and fracture mode of porcelain-veneered Y-TZP crown cores and monolithic lithium disilicate glass-ceramic crowns.

The kinetics and homing characteristics of T-cell responses in humans after mucosal immunizations have not been well characterized. Therefore, we have investigated the magnitude and duration of such responses as well as the homing receptor expression of antigen-specific peripheral blood T cells by using an oral model vaccine, i.e., the live, attenuated Salmonella enterica serovar Typhi vaccine (Ty21a). Eight volunteers were each given three doses of the vaccine 2 days apart, and blood samples, from which CD4 ؉ and CD8 ؉ T cells were selected by the use of magnetic beads, were collected before vaccination and at regular intervals thereafter. To purify the potentially antigen-specific gut-homing T cells, CD45RA؊ integrin ␤ 7 ؉ cells were further sorted by flow cytometry. The sorted cells were then stimulated in vitro with the serovar Typhi vaccine strain, and the proliferation of cells and the cytokine production were measured. Following vaccination, there was a large increase in both the proliferation of and the gamma interferon (IFN-␥) production by blood T cells stimulated with the vaccine strain. The responses were seen among both CD4؉ and CD8 ؉ T cells, although the CD8 ؉ cells produced the largest amounts of IFN-␥. Peak responses were seen 7 to 14 days after the onset of vaccination. Furthermore, most of the IFN-␥ produced by both CD4؉ and CD8 ؉ cells emanated from cells with the potential to home to mucosal tissues, as the integrin ␤ 7 -expressing memory T cells produced around 10-fold more IFN-␥ than the remaining populations. In conclusion, we demonstrate that oral vaccination with a live oral bacterial vaccine induces antigen-specific CD4؉ and CD8 ؉ memory T cells, almost all of which express the gut-homing integrin ␤ 7 .Although the kinetics and homing characteristics of B-cell responses in humans after mucosal immunizations have been extensively studied, mucosal T-cell responses after oral vaccination have been investigated in only a few studies (9,19,22,23,24). Oral vaccination with the attenuated Salmonella enterica serovar Typhi strains Ty21a (Vivotif) and CVD908 has been shown to induce proliferative responses, production of gamma interferon (IFN-␥), and antibody-dependent antibacterial activity by peripheral blood lymphocytes in the majority of vaccinated subjects at 3 weeks postvaccination (19,21). Furthermore, a more recent report demonstrated that volunteers vaccinated with two doses of the CVD908 strain developed CD8ϩ cytotoxic-T-lymphocyte precursors, which after a short in vitro expansion were capable of killing serovar Typhiinfected cells (18). Except in that study, the relative contributions of CD4 ϩ and CD8 ϩ T cells in the response to oral vaccinations have not been investigated and the kinetics and homing patterns of the T-cell responses have not been evaluated in any detail.A central event in the homing of lymphocytes to different tissues is the adhesion of lymphocytes by means of surfacehoming receptors to binding structures, so-called addressins, on endothelial cells. Because some of...

Receptor-dependent productive uptake of GLP1-conjugated antisense oligonucleotides occurs selectively in pancreatic β-cells.

Background Duchenne muscular dystrophy (DMD) is a fatal disease for which no cure is available. Clinical trials have shown to be largely underpowered due to inter-individual variability and noisy outcome measures. The availability of biomarkers able to anticipate clinical benefit is highly needed to improve clinical trial design and facilitate drug development.Methods In this study, we aimed to appraise the value of protein biomarkers to predict prognosis and monitor disease progression or treatment outcome in patients affected by DMD. We collected clinical data and 303 blood samples from 157 DMD patients in three clinical centres; 78 patients contributed multiple blood samples over time, with a median follow-up time of 2 years. We employed linear mixed models to identify biomarkers that are associated with disease progression, wheelchair dependency, and treatment with corticosteroids and performed survival analysis to find biomarkers whose levels are associated with time to loss of ambulation. Results Our analysis led to the identification of 21 proteins whose levels significantly decrease with age and nine proteins whose levels significantly increase. Seven of these proteins are also differentially expressed in non-ambulant patients, and three proteins are differentially expressed in patients treated with glucocorticosteroids. Treatment with corticosteroids was found to partly counteract the effect of disease progression on two biomarkers, namely, malate dehydrogenase 2 (MDH2, P = 0.0003) and ankyrin repeat domain 2 (P = 0.0005); however, patients treated with corticosteroids experienced a further reduction on collagen 1 serum levels (P = 0.0003), especially following administration of deflazacort. A time to event analysis allowed to further support the use of MDH2 as a prognostic biomarker as it was associated with an increased risk of wheelchair dependence (P = 0.0003). The obtained data support the prospective evaluation of the identified biomarkers in natural history and clinical trials as exploratory biomarkers. Conclusions We identified a number of serum biomarkers associated with disease progression, loss of ambulation, and treatment with corticosteroids. The identified biomarkers are promising candidate prognostic and surrogate biomarkers, which may support drug developers if confirmed in prospective studies. The serum levels of MDH2 are of particular interest, as they correlate with disease stage and response to treatment with corticosteroids, and are also associated with the risk of wheelchair dependency and pulmonary function.

Lipid nanoparticles (LNPs) are the most clinically advanced delivery system for RNA-based drugs but have predominantly been investigated for intravenous and intramuscular administration. Subcutaneous administration opens the possibility of patient self-administration and hence long-term chronic treatment that could enable messenger RNA (mRNA) to be used as a novel modality for protein replacement or regenerative therapies. In this study, we show that subcutaneous administration of mRNA formulated within LNPs can result in measurable plasma exposure of a secreted protein. However, subcutaneous administration of mRNA formulated within LNPs was observed to be associated with dose-limiting inflammatory responses. To overcome this limitation, we investigated the concept of incorporating aliphatic ester prodrugs of anti-inflammatory steroids within LNPs, i.e., functionalized LNPs to suppress the inflammatory response. We show that the effectiveness of this approach depends on the alkyl chain length of the ester prodrug, which determines its retention at the site of administration. An unexpected additional benefit to this approach is the prolongation observed in the duration of protein expression. Our results demonstrate that subcutaneous administration of mRNA formulated in functionalized LNPs is a viable approach to achieving systemic levels of therapeutic proteins, which has the added benefits of being amenable to self-administration when chronic treatment is required.