Contribution of the Dynamic Intestinal Absorption Model (Diamod) to the Development of a Patient-Centric Drug Formulation.

Compound X is a weak basic drug targeting the early stages of Parkinson's disease, for which a theoretical risk assessment has indicated that elevated gastric pH conditions could potentially result in reduced plasma concentrations. Different in vitro dissolution methodologies varying in level of complexity and a physiologically based pharmacokinetic (PBPK) absorption model demonstrated that the dissolution, solubility, and intestinal absorption of compound X was indeed reduced under elevated gastric pH conditions. These observations were confirmed in a crossover pharmacokinetic study in Beagle dogs. As a result, the development of a formulation resulting in robust performance that is not sensitive to the exposed gastric pH levels is of crucial importance. The dynamic intestinal absorption MODel (Diamod), an advanced in vitro gastrointestinal transfer tool that allows to study the gastrointestinal dissolution and interconnected permeation of drugs, was selected as an in vitro tool for the formulation optimization activities given its promising predictive capacity and its capability to generate insights into the mechanisms driving formulation performance. Different pH-modifiers were screened for their potential to mitigate the pH-effect by decreasing the microenvironmental pH at the dissolution surface. Finally, an optimized formulation containing a clinically relevant dose of the drug and a functional amount of the selected pH-modifier was evaluated for its performance in the Diamod. This monolayer tablet formulation resulted in rapid gastric dissolution and supersaturation, inducing adequate intestinal supersaturation and permeation of compound X, irrespective of the gastric acidity level in the stomach. In conclusion, this study describes the holistic biopharmaceutics approach driving the development of a patient-centric formulation of compound X.

Risk Factors Associated with Failure and Technical Complications of Implant-Supported Single Crowns: A Retrospective Study.

Background and Objectives: Implant-supported single crowns have become a routine approach for the replacement of missing single teeth, being considered as one of the most common ways of rehabilitation when adjacent teeth are healthy. The present retrospective study aimed to investigate the risk factors possibly associated with failure and technical complications of implant-supported single crowns and their supporting implants. Materials and Methods: Patients treated at one faculty (2009-2019) were considered for inclusion. Complications investigated included ceramic fracture/chipping, crown loss of retention/mobility, crown failure/fracture, loosening/loss/fracture of prosthetic screw, and implant failure/fracture. Any condition/situation that led to the removal/replacement of crowns (implant failure not included) was considered prosthesis failure. Univariate/multivariate Cox regression models were used to evaluate the associations between clinical covariates and failure. Results: 278 patients (358 crowns) were included. Mean ± SD follow-up was 56.5 ± 29.7 months. Seven implants (after a mean of 76.5 ± 43.7 months) and twenty crowns (21.3 ± 23.5 months) failed. The cumulative survival rate (CSR) for crowns was 93.5% after 5, remaining at 92.2% between 6 and 11 years. The most common reasons for crown failure were porcelain large fracture (n = 6), crown repeatedly loose (n = 6), and porcelain chipping (n = 5). Men and probable bruxism were identified in the Cox regression model as being associated with crown failure. The most common observed technical complications were mobility of the crown and chipping of the ceramic material, with the latter being observed even in crowns manufactured of monolithic zirconia. Cases with at least one technical complication (not considering loss of screw hole sealing) were more common among probable bruxers than in non-bruxers (p = 0.002). Cases of ceramic chipping were more common among bruxers than in non-bruxers (p = 0.014, log-rank test). Conclusions: Probable bruxism and patient's sex (men) were factors associated with a higher risk of failure of implant-supported single crowns.

The Dynamic Intestinal Absorption Model (Diamod®), an in vitro tool to study the interconnected kinetics of gastrointestinal solubility, supersaturation, precipitation, and intestinal permeation processes of oral drugs.

This study aimed at developing the Diamod® as a dynamic gastrointestinal transfer model with physically interconnected permeation. The Diamod® was validated by studying the impact of the intraluminal dilution of a cyclodextrin-based itraconazole solution and the negative food effect for indinavir sulfate for which clinical data are available demonstrating that the systemic exposure was strongly mediated by interconnected solubility, precipitation, and permeation processes. The Diamod® accurately simulated the impact of water intake on the gastrointestinal behavior of a Sporanox® solution. Water intake significantly decreased the duodenal solute concentrations of itraconazole as compared to no intake of water. Despite this duodenal behavior the amount of permeated itraconazole was not affected by water intake as observed in vivo. Next to this, the Diamod® accurately simulated the negative food effect for indinavir sulfate. Different fasted and fed state experiments demonstrated that this negative food effect was mediated by an increased stomach pH, entrapment of indinavir in colloidal structures and the slower gastric emptying of indinavir under fed state conditions. Therefore, it can be concluded that the Diamod® is a useful in vitro model to mechanistically study the gastrointestinal performance of drugs.

Split-marker-mediated genome editing improves homologous recombination frequency in the CTG clade yeast Candida intermedia.

Genome-editing toolboxes are essential for the exploration and exploitation of nonconventional yeast species as cell factories, as they facilitate both genome studies and metabolic engineering. The nonconventional yeast Candida intermedia is a biotechnologically interesting species due to its capacity to convert a wide range of carbon sources, including xylose and lactose found in forestry and dairy industry waste and side-streams, into added-value products. However, possibilities of genetic manipulation have so far been limited due to lack of molecular tools for this species. We describe here the development of a genome editing method for C. intermedia, based on electroporation and gene deletion cassettes containing the Candida albicans NAT1 dominant selection marker flanked by 1000 base pair sequences homologous to the target loci. Linear deletion cassettes targeting the ADE2 gene originally resulted in <1% targeting efficiencies, suggesting that C. intermedia mainly uses nonhomologous end joining for integration of foreign DNA fragments. By developing a split-marker based deletion technique for C. intermedia, we successfully improved the homologous recombination rates, achieving targeting efficiencies up to 70%. For marker-less deletions, we also employed the split-marker cassette in combination with a recombinase system, which enabled the construction of double deletion mutants via marker recycling. Overall, the split-marker technique proved to be a quick and reliable method for generating gene deletions in C. intermedia, which opens the possibility to uncover and enhance its cell factory potential.