Inhibitory Effect of Multivalent Rhamnobiosides on Recombinant Horseshoe Crab Plasma Lectin Interactions with Pseudomonas aeruginosa PAO1.

To evaluate the molecular interaction of recombinant horseshoe crab plasma lectin (rHPL) with Pseudomonas aeruginosa PAO1, multivalent rhamnobioside derivatives were designed. Eight rhamnoclusters with three or four α(1-3)-rhamnobiosides attached to different central cores, such as methyl gallate, pentaerythritol, and N-Boc Tris, through either an ethylene glycol or a tetraethylene glycol linker, were assembled in two consecutive azide-alkyne cycloaddition click reactions. The synthetic method embraced the preparation of two α(1-3)-rhamnobiosides with different linker arms and their conjugation, in stoichiometric or substoichiometric amounts, to propargyl ether-functionalized tri- or tetravalent scaffolds. A divalent derivative and two self-assembling rhamnobiosides were also prepared. The different architectures and valences of the rhamnoclusters provided an opportunity to evaluate the impact of topology and valency on the binding properties toward rHPL. Inhibitory ELISA data showed that all covalently linked rhamnoclusters could inhibit P. aeruginosa PAO1 recognition activity of rHPL with high efficacy. Trivalent rhamnobiosides showed a stronger inhibitory effect on P. aeruginosa PAO1 binding, and the more flexible clusters on a pentaerythritol or a Tris core were superior to the less flexible methyl gallate-based clusters. Interestingly, the length of the linker arms had a very low impact on the binding ability of the rhamnoclusters. Herein, the two trivalent derivatives on an N-Boc protected Tris central core were the best inhibitors. The self-assembling amphiphilic rhamnobioside derivatives were found to display no multivalent effect.

Antidepressants cause foot detachment from substrate in five species of marine snail.

Active Pharmaceutical Ingredients (APIs) are released into aquatic ecosystems through discharged sewage wastewater. Antidepressants are among those APIs often detected in wastewater effluent and have been recently reported to cause foot detachment from the substrate in freshwater snails. We tested the effects of four commonly prescribed antidepressants {fluoxetine ("Prozac"), fluvoxamine ("Luvox"), venlafaxine ("Effexor"), and citalopram ("Celexa") on adhesion to the substrate in five species of marine snails, three from the Pacific coast (Chlorostoma funebralis, Nucella ostrina, Urosalpinx cinerea) and two species from the Atlantic coast (Tegula fasciatus and Lithopoma americanum) of North America representing three different gastropod families. All antidepressants tested induced foot detachment from the substrate in all snail species in a mainly dose-dependent manner (p < 0.04-0.00000001). The lowest LOECs (lowest observed effect concentration) for antidepressants and snails were recorded for Lithopoma in 43.4 µg/L (100 nM) fluvoxamine and Chlorostoma in 157 µg/L (500 nM) venlafaxine and 217 µg/L (500 nM) fluvoxamine. The trochids and turbinids were 2-10× more sensitive to the antidepressants than the muricids. Latency to detachment was also dose dependent, with the fastest average times to detach seen in Chlorostoma and Lithopoma (7.33 and 13.16 min respectively in 3.13 mg/L venlafaxine). The possible physiological mechanisms regulating antidepressant-induced foot detachment in marine snails and the possible ecological consequences are discussed.

Inhibition of endothelial/smooth muscle cell contact loss by the investigational angiopoietin-2 antibody MEDI3617.

A tumor's dependence on angiogenesis for survival and growth has led to the advancement of a variety of blood vessel directed anticancer treatment strategies. Overexpression of angiopoietin-2 (Ang-2) in tumor vasculature and its crucial role in angiogenesis, i.e. the destabilization of endothelial/peri-endothelial cell interactions, now raises the possibility of additional novel anti-angiogenic therapeutics. The present study utilized a co-culture sphere model to (i) demonstrate the destabilizing effect of Ang-2 on endothelial/smooth muscle cell contact and (ii) evaluate the impact of the investigational Ang-2 antibody MEDI3617 on endothelial/smooth muscle cell dissociation. Real time imaging of spheres showed both exogenous Ang-2 and PMA induced endogenous Ang-2 secretion resulted in sphere destabilization (loss of endothelial cells from smooth muscle cell core). The presence of MEDI3617 inhibited this process. To assess the anti-angiogenic potential of MEDI3617 in vivo, nude mice were injected intradermally with human renal cell carcinoma cells (Caki-1, Caki-2) and the number of blood vessels induced over a 3 day period was scored. MEDI3617 (2, 10, 20 mg/kg) significantly reduced the initiation of blood vessels for both tumor models at all doses investigated. These data indicate that MEDI3617 treatment significantly impairs the initiation of angiogenesis by inhibiting the Ang-2 mediated disruption of endothelial/muscle cell interaction associated with blood vessel destabilization and thereby reduces tumor cell induced angiogenesis. The results support the notion that targeting the angiopoietin/Tie2 axis may offer novel anti-angiogenic strategies for cancer treatment.