TY - JOUR AB - Cellular vesicles (CVs) have been proposed as alternatives to exosomes for targeted drug delivery. CVs, prepared from human embryonic kidney 293 cells (HEK-293), C57BL/6 mouse B16F10 skin melanoma cells (B16F10), and immortalized human cerebral microvascular endothelial cells (hCMEC/D3) by liposome technology methods, were characterized for morphology, cytotoxicity, and cell uptake properties. CV brain-targeting potential was evaluated in vitro on the hCMEC/D3 blood-brain barrier (BBB) model, and in vivo/ex vivo. CV sizes were between 135 and 285 nm, and the ζ-potential was negative. The dehydration-rehydration method conferred highest calcein loading and latency to CVs compared with other methods. The increased calcein leakage from CVs when compared with liposomes indicated their poor integrity, which was increased by pegylation. The in vivo results confirmed lower liver uptake by PEG-CVs (compared with nonpegylated) proving that the calcein integrity test is useful for prediction of CV biodistribution, as used for liposomes. The cell uptake of homologous origin CVs was not always higher compared with that of non-homologous. Nevertheless, CVs from hCMEC/D3 demonstrated the highest BBB permeability (in vitro) compared with OX-26 targeted liposomes, and brain localization (in vivo). CVs from hCMEC/D3 cells grown in different media demonstrated decreased interaction with brain cells and brain localization. Significant differences in proteome of the two latter CV types were identified by proteomics, suggesting a potential methodology for identification of organotropism-determining CV components. AU - Marazioti, A.* AU - Papadia, K.* AU - Kannavou, M.* AU - Spella, M.* AU - Basta, A.* AU - de Lastic, A.L.* AU - Rodi, M.* AU - Mouzaki, A.* AU - Samiotaki, M.* AU - Panayotou, G.* AU - Stathopoulos, G.T. AU - Antimisiaris, S.G.* C1 - 56876 C2 - 47408 SP - 772-785 TI - Cellular vesicles: New insights in engineering methods, interaction with cells and potential for brain targeting. JO - J. Pharmacol. Exp. Ther. VL - 370 IS - 3 PY - 2019 SN - 0022-3565 ER - TY - JOUR AB - G-quadruplexes (G4) are nucleic acid secondary structures frequently assumed by G-rich sequences located mostly at telomeres and proto-oncogenes promoters. Recently, we identified, in canine KIT (v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog) promoter, two G-rich sequences able to fold into G4: d_kit1 and d_kit2_A16. In this study, an anthraquinone (AQ1) and an anthracene derivative (AN6), known to stabilize the G4 structures of the corresponding human h_kit1 and h_kit2, were tested on the canine G4 and in two canine mast cell tumor (MCT) cell lines (C2 and NI-1) to verify their capability to down-regulate KIT expression. The cytotoxicity of AQ1 and AN6 was determined using the Alamar Blue test; also the constitutive expression of KIT and other proto-oncogenes containing G4 structures in their promoter (BCL2, VEGF alpha, VEGFR2, KRAS, and TERT) was assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Then the time- and dose-dependent effects of both ligands on target gene expression were assessed by qRT-PCR. All target genes were constitutively expressed up to 96 hours of culture. Both ligands decreased KIT mRNA levels and c-kit protein amount, and AN6 was comparatively fairly more effective. DNA interaction studies and a dual-luciferase gene reporter assay performed on a noncancerous canine cell line (Madin-Darby Canine Kidney cells) proved that this down-regulation was the result of the interaction of AN6 with KIT proximal promoter. Interestingly, our results only partially overlap with those previously obtained in human cell lines, where AQ1 was found as the most effective compound. These preliminary data might suggest AN6 as a promising candidate for the selective targeting of canine KIT-dependent tumors. AU - Zorzan, E.* AU - Da Ros, S.* AU - Giantin, M.* AU - Zorro Shahidian, L. AU - Guerra, G.* AU - Palumbo, M.* AU - Sissi, C.* AU - Dacasto, M.* C1 - 54442 C2 - 45609 CY - 9650 Rockville Pike, Bethesda, Md 20814-3995 Usa SP - 461-472 TI - Targeting canine KIT promoter by candidate DNA G-quadruplex ligands. JO - J. Pharmacol. Exp. Ther. VL - 367 IS - 3 PB - Amer Soc Pharmacology Experimental Therapeutics PY - 2018 SN - 0022-3565 ER - TY - JOUR AU - Beck-Speier, I. AU - Dayal, N. AU - Karg, E.W. AU - Maier, K.L. AU - Schumann, G. AU - Semmler, M. AU - Koelsch, S.M.* C1 - 3551 C2 - 23448 SP - 843-851 TI - Oxymetazoline inhibits proinflammatory reactions: Effect on arachidonic acid-derived metabolites. JO - J. Pharmacol. Exp. Ther. VL - 316 PY - 2006 SN - 0022-3565 ER - TY - JOUR AU - Rozman, K.K. AU - Doull, J.* C1 - 21866 C2 - 20090 SP - 663-668 TI - The Role of Time as a Quantifiable Variable of Toxicity and the Experimental Conditions When Haber's c x t Product Can Be Observed : Implications for Therapeutics. JO - J. Pharmacol. Exp. Ther. VL - 296 PY - 2001 SN - 0022-3565 ER - TY - JOUR AB - Distribution, metabolism, and excretion of monochloroacetic acid (MCA) were examined in adult male rats at a subtoxic (10 mg/kg) and a toxic (75 mg/kg) dose. Rats were injected i.v. with [14C]MCA and housed individually. Urine and feces were collected. Animals were euthanized at different time intervals after dosing and tissues procured. Radioactivity in aliquots showed very rapid distribution of MCA to tissues. Concentrations of MCA in plasma, liver, heart, lungs, and brown fat paralleled each other, whereas those in brain and thymus did not. There was no dose proportionality in tissue concentrations. Elimination of MCA from plasma required modeling by two compartments. Most of the radioactivity found in plasma was parent MCA. Elimination rate constant (K(10)) and distribution rate constant (K(12)) were greatly reduced at the toxic dose. Elimination of the toxic dose was further retarded due to increased retention of MCA in the peripheral compartment as indicated by increased mean residence times in most tissues. A very large fraction of dose was found in the gastrointestinal tract, almost all of which was reabsorbed. Attempts to reduce toxicity by blocking the enterohepatic circulation with activated charcoal or cholestyramine failed. Radioactivity found in bile was associated with one metabolite more polar than the parent compound. A very large fraction of dose (73 and 59%) was found in urine, 55 to 68% of which was parent MCA. The rate-determining step in the toxicity of MCA was identified as its detoxification by the liver. A therapeutic approach in MCA intoxications is suggested. AU - Saghir, S.A.* AU - Fried, K.* AU - Rozman, K.K. C1 - 21865 C2 - 20091 SP - 617-627 TI - Kinetics of monochloroacetic acid in adult male rats after intravenous injection of a subtoxic and a toxic dose. JO - J. Pharmacol. Exp. Ther. VL - 296 IS - 2 PB - Amer. Soc. Pharmacology Experimental Therapeutics PY - 2001 SN - 0022-3565 ER -