Screening and Characterisation of Antimicrobial Properties of Semisynthetic Betulin Derivatives

Peer reviewe

Antimicrobial activity of the marine alkaloids, clathrodin and oroidin, and their synthetic analogues

Peer reviewe

Is oxygenation related to the decomposition of organic matter in cryoconite holes?

Cryoconite is a sediment occurring on glacier surfaces worldwide which reduces ice albedo and concentrates glacier surface meltwater into small reservoirs called cryoconite holes. It consists of mineral and biogenic matter, including active microorganisms. This study presents an experimental insight into the influence of sediment oxygenation on the cryoconite ability to produce and decomposition of organic matter. Samples were collected from five glaciers in the Arctic and the European mainland. Cryoconite from three glaciers was incubated in stagnant and mechanically mixed conditions to imitate inter-hole water–sediment mixing by meltwater occurring on glaciers in Northern Hemisphere, and its effect on oxygen profiles and organic matter content. Moreover, we investigated short-term changes of oxygen conditions in cryoconite from four glaciers in illuminated and dark conditions. An anaerobic zone was present or approaching zero oxygen in all illuminated cryoconite samples, varying in depth depending on the origin of cryoconite: from 1500 µm from Steindalsbreen (Scandinavian Peninsula) and Forni Glacier (The Alps) to 3100 µm from Russell Glacier and Longyearbreen (Arctic) after incubation. Organic matter content varied between glaciers from 6.11% on Longyearbreen to 16.36% on Russell Glacier. The mixed sediment from the Forni Glacier had less organic matter than stagnant, the sediment from Longyearbreen followed this trend, but the difference was not statistically significant, while the sediment from Ebenferner did not differ between groups. Our results have implications for the understanding of biogeochemical processes on glacier surfaces, the adaptation of organisms to changing physical conditions due to abrupt sediment mixing, but also on the estimation of productivity of supraglacial systems

The retinoid agonist Tazarotene promotes angiogenesis and wound healing

Therapeutic angiogenesis is a major goal ofregenerative medicine, but no clinically approved small molecule exists that enhancesnew blood vessel formation. Here we show, using a phenotype-driven high-content imaging screen of an annotated chemical library of 1280 bioactive small molecules, that the retinoid agonist Tazarotene, enhances in vitroangiogenesis, promoting branching morphogenesis, and tubule remodeling. The pro-angiogenic phenotype is mediated by Retinoic Acid Receptor (RAR) but not Retinoic X Receptor(RXR) activation, and is characterized by secretion of the pro-angiogenic factors Hepatocyte Growth Factor (HGF), Vascular Endothelial Growth Factor (VEGFA), Plasminogen Activator, Urokinase (PLAU) and Placental Growth Factor (PGF), and reduced secretion of the antiangiogenic factor Pentraxin-3 (PTX3) from adjacent fibroblasts. In vivo, Tazarotene enhanced the growth of mature and functional microvessels in Matrigel implants and wound healing models, and increased blood flow. Notably, in ear punch wound healing model, Tazarotene promoted tissue repair characterized by rapid ear punch closure with normal-appearing skin containing new hair follicles, and maturing collagen fibers. Our study suggests that Tazarotene, an FDA-approved small molecule, could be potentially exploited for therapeutic applications in neovascularization and wound healing

A Novel High Content Angiogenesis Assay Reveals That Lacidipine, L-Type Calcium Channel Blocker, Induces In Vitro Vascular Lumen Expansion

Angiogenesis is a critical cellular process toward establishing a functional circulatory system capable of delivering oxygen and nutrients to the tissue in demand. In vitro angiogenesis assays represent an important tool for elucidating the biology of blood vessel formation and for drug discovery applications. Herein, we developed a novel, high content 2D angiogenesis assay that captures endothelial morphogenesis’s cellular processes, including lumen formation. In this assay, endothelial cells form luminized vascular-like structures in 48 h. The assay was validated for its specificity and performance. Using the optimized assay, we conducted a phenotypic screen of a library containing 150 FDA-approved cardiovascular drugs to identify modulators of lumen formation. The screening resulted in several L-type calcium channel blockers being able to expand the lumen space compared to controls. Among these blockers, Lacidipine was selected for follow-up studies. We found that the endothelial cells treated with Lacidipine showed enhanced activity of caspase-3 in the luminal space. Pharmacological inhibition of caspase activity abolished the Lacidipine-enhancing effect on lumen formation, suggesting the involvement of apoptosis. Using a Ca2+ biosensor, we found that Lacipidine reduces the intracellular Ca2+ oscillations amplitude in the endothelial cells at the early stage, whereas Lacidipine blocks these Ca2+ oscillations completely at the late stage. The inhibition of MLCK exhibits a phenotype of lumen expansion similar to that of Lacidipine. In conclusion, this study describes a novel high-throughput phenotypic assay to study angiogenesis. Our findings suggest that calcium signalling plays an essential role during lumen morphogenesis. L-type Ca2+ channel blockers could be used for more efficient angiogenesis-mediated therapies

Antimicrobial activity of the marine alkaloids, clathrodin and oroidin, and their synthetic analogues

Marine organisms produce secondary metabolites that may be valuable for the development of novel drug leads as such and can also provide structural scaffolds for the design and synthesis of novel bioactive compounds. The marine alkaloids, clathrodin and oroidin, which were originally isolated from sponges of the genus, Agelas, were prepared and evaluated for their antimicrobial activity against three bacterial strains (Enterococcus faecalis, Staphylococcus aureus and Escherichia coli) and one fungal strain (Candida albicans), and oroidin was found to possess promising Gram-positive antibacterial activity. Using oroidin as a scaffold, 34 new analogues were designed, prepared and screened for their antimicrobial properties. Of these compounds, 12 exhibited >80% inhibition of the growth of at least one microorganism at a concentration of 50 µM. The most active derivative was found to be 4-phenyl-2-aminoimidazole 6h, which exhibited MIC$_{90}$ (minimum inhibitory concentration) values of 12.5 µM against the Gram-positive bacteria and 50 µM against E. coli. The selectivity index between S. aureus and mammalian cells, which is important to consider in the evaluation of a compound\u27s potential as an antimicrobial lead, was found to be 2.9 for compound 6h