Active Intracellular Delivery of a Cas9/sgRNA Complex Using Ultrasound‐Propelled Nanomotors

Direct and rapid intracellular delivery of a functional Cas9/sgRNA complex using ultrasound‐powered nanomotors is reported. The Cas9/sgRNA complex is loaded onto the nanomotor surface through a reversible disulfide linkage. A 5 min ultrasound treatment enables the Cas9/sgRNA‐loaded nanomotors to directly penetrate through the plasma membrane of GFP‐expressing B16F10 cells. The Cas9/sgRNA is released inside the cells to achieve highly effective GFP gene knockout. The acoustic Cas9/sgRNA‐loaded nanomotors display more than 80 % GFP knockout within 2 h of cell incubation compared to 30 % knockout using static nanowires. More impressively, the nanomotors enable highly efficient knockout with just 0.6 nm of the Cas9/sgRNA complex. This nanomotor‐based intracellular delivery method thus offers an attractive route to overcome physiological barriers for intracellular delivery of functional proteins and RNAs, thus indicating considerable promise for highly efficient therapeutic applications.     

Ageing of spherulites of an industrial relevant aromatic amine derivative

Polycrystalline spherulites of an aromatic amine derivative have been precipitated in a batch process by pH‐shift with hydrochloric acid from stirred aqueous solutions. The time dependent behaviour of the spherulites has been studied during crystallization in the temperature range from 5 °C to 60 °C. Cake resistance values have been obtained from batch filtration tests performed at 2 bar pressure difference at different stages of the crystallization process. The FBRM mean chord length of the crystals decreases with time as a result of crystal ageing into plate‐like crystals. The rate of the ageing process increases with temperature. XRD‐studies show no significant differences in the crystal structure during the ageing process, and the mechanism of the transformation was not established. The filterability of aromatic amine crystals deteriorates as the crystallization progresses. The decrease in the filterability is attributed to the appearance of small plate‐like crystals and a change in the interaction between the crystal surface and the solution, during the ageing of the particles. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Formation and ageing of L‐glutamic acid spherulites

Polycrystalline spherulites of L‐glutamic acid have been crystallized by pH‐shift precipitation from stirred aqueous solutions. The time dependent behaviour of the spherulites has been studied during the crystallization process and batch filtration tests have been performed. It has been shown that the FBRM mean chord length of the investigated spherulites decreases in the course of time. The fact that the size reduction progresses faster at higher temperature and the solubility of resuspended polycrystalline particles decreasing with time, implies an ageing mechanism to be responsible for the observed changes in the particle size. It has been shown that the surface area decreases with time, ruling out particle breakage as a possible explanation for the decrease in particle size. XRD and Raman studies of L‐glutamic acid, however, show only marginal differences in the crystalline structure of particles obtained from different time stages. The ageing may occur due to several different mechanisms like phase transformation and Ostwald ripening. L‐glutamic acid spherulites after 3 h exhibit a 3‐fold higher value for the cake resistance as compared to particles after 0.5 h. However, particles obtained after 22 h exhibit an 8‐fold lower cake resistance as compared to the initially obtained spherulites, The increase in the cake resistance is attributed to the appearance of small plate‐like crystals and a change in the interaction between the crystal surface and the solution. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Pharmacological Insights into Halophyte Bioactive Extract Action on Anti-Inflammatory,Pain Relief and Antibiotics-Type Mechanisms

The pharmacological activities in bioactive plant extracts play an increasing role in sustainable resources for valorization and biomedical applications. Bioactive phytochemicals, including natural compounds, secondary metabolites and their derivatives, have attracted significant attention for use in both medicinal products and cosmetic products. Our review highlights the pharmacological mode-of-action and current biomedical applications of key bioactive compounds applied as anti-inflammatory, bactericidal with antibiotics effects, and pain relief purposes in controlled clinical studies or preclinical studies. In this systematic review, the availability of bioactive compounds from several salt-tolerant plant species, mainly focusing on the three promising species Aster tripolium, Crithmum maritimum and Salicornia europaea, are summarized and discussed. All three of them have been widely used in natural folk medicines and are now in the focus for future nutraceutical and pharmacological applications.