Synthetic Biology of Cannabinoids and Cannabinoid Glucosides in Nicotiana benthamiana and Saccharomyces cerevisiae.

Phytocannabinoids are a group of plant-derived metabolites that display a wide range of psychoactive as well as health-promoting effects. The production of pharmaceutically relevant cannabinoids relies on extraction and purification from cannabis (Cannabis sativa) plants yielding the major constituents, Δ9-tetrahydrocannabinol and cannabidiol. Heterologous biosynthesis of cannabinoids in Nicotiana benthamiana or Saccharomyces cerevisiae may provide cost-efficient and rapid future production platforms to acquire pure and high quantities of both the major and the rare cannabinoids as well as novel derivatives. Here, we used a meta-transcriptomic analysis of cannabis to identify genes for aromatic prenyltransferases of the UbiA superfamily and chalcone isomerase-like (CHIL) proteins. Among the aromatic prenyltransferases, CsaPT4 showed CBGAS activity in both N. benthamiana and S. cerevisiae. Coexpression of selected CsaPT pairs and of CHIL proteins encoding genes with CsaPT4 did not affect CBGAS catalytic efficiency. In a screen of different plant UDP-glycosyltransferases, Stevia rebaudiana SrUGT71E1 and Oryza sativa OsUGT5 were found to glucosylate olivetolic acid, cannabigerolic acid, and Δ9-tetrahydrocannabinolic acid. Metabolic engineering of N. benthamiana for production of cannabinoids revealed intrinsic glucosylation of olivetolic acid and cannabigerolic acid. S. cerevisiae was engineered to produce olivetolic acid glucoside and cannabigerolic acid glucoside.

Comparison of statistical methods for predicting penetration capacity of drugs into human breast milk using physicochemical, pharmacokinetic and chromatographic descriptors.

In silico methods are often used for predicting pharmacokinetic properties of drugs due to their simplicity and cost-effectiveness. This study evaluates the penetration of 83 active pharmaceutical ingredients into human breast milk with an experimental milk-to-plasma ratio (M/P) obtained from the literature. Multiple linear regression (MLR), partial least squares (PLS) and random forest (RF) regression methods were compared to uncover the relationship between physicochemical, pharmacokinetic and membrane crossing properties of active pharmaceutical ingredients (APIs) using their rapid reference measurement value (Rf values), thin-layer chromatography (TLC) data from albumin-impregnated plates. Molecular descriptors of APIs proven to be important for their crossing into breast milk, including protein binding, ionisation state and lipophilicity and TLC data, have been included in the development of the prediction models. The best regression results were achieved by MLR (r 2 = 0.83 and r 2 = 0.86, n = 28) and RF (r 2 = 0.85, n = 58). In addition, the discriminant function analysis (DFA) was performed on acidic, basic and neutral drugs separately and showed a prediction accuracy of 93%, with M/P included as the discriminating variable.

[Minimally invasive interventions for acute cholecystitis complicated by mechanical jaundice in elderly and senile patients].

Results of the diagnosis and treatment of 165 patients with acute cholecystitis complicated by mechanical jaundice in elderly and senile age were analyzed. The method of endoscopic retrograde papillocholangiography used in 153 patients with acute cholecystitis complicated by mechanical jaundice is thought to be the most informative method of investigation of the bile ducts which allows determination of the exact cause and the level of disturbance of their patency. In 138 (83.6%) patients the cause of these complications was choledocholithiasis, in 18 (10.9%) isolated scarry strictures of the terminal part of the common bile duct and major duodenal papilla and in 9 patients (5.5%) a combination of these two diseases. Purulent cholangitis developed in 61 patients. Operations were fulfilled in 122 patients at the second stage, in 38 patients it was laparoscopic cholecystectomy. The differentiated medico-diagnostic strategy for acute cholecystitis complicated by mechanical jaundice using a two-stage method of decompression and sanitation of the bile ducts is the most rational method of treatment of elderly and senile patients.