Tajemství vládnutí spočívá totiž v tom, jak spojit víru ve vlastní neomylnost se schopností učit se z minulých chyb.

George Orwell

Výzkum

Electrochemical and Spectrometric Study of GFP-AZT Interaction

Highly active antiretroviral therapy (HAART) was introduced in 1996 and revolutionized the existing treatment of patients infected by human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS). Currently, this type of treatment is called also as a combination antiretroviral therapy (cART) [1]. With the arrival of this therapy the incidence of opportunistic infections and premature death was significantly reduced [2]. Before the year 1996 the median of survival of young infected people was 7 years after the diagnosis, while currently it is 35 years [3]. This treatment consists of a combination of protease inhibitors (PI), nucleoside reverse transcriptase inhibitors (NRTI), and/or non-nucleoside reverse transcriptase inhibitors (NNRTI) [4]. The group of NRTI includes also azidothymidine (3'-Azido-3'-deoxythymidine, Zidovudine, AZT) and together with another drug from this class is administered as a part of HAART particularly in the treatment of HIV-1. This is the first compound that has been approved for the treatment of HIV and is still an integral part of systemic therapy [5]. AZT, as well as drugs belonging to NRTI, requires activation by phosphorylation [6], then it is followed by the end of termination and by the blocking of nucleotide-binding site for HIV-1 reverse transcriptase [7-9]. Besides this mechanism the using of this treatment also leads to the inhibition of gamma-polymerase resulting in decline in the occurrence of mtDNA [10,11]. Despite the fact that thanks to the use of AZT and other drugs under HAART the suppression of viral replication occurs in a long term, this effect is redeemed by a number of side effects, which follow exactly the decrease of the mtDNA and thus lead to mitochondrial toxicity that leads to a liver failure and a lactic acidosis [10]. Side effects associated with HAART therapy include anaemia, cardiomyopathy, gastrointestinal disturbance, drug-induced hypersensitivity, nephrotoxicity, pancreatitis, ototoxicity and others [6]. AZT, compared to other drugs from NRTI group, shows markedly less inhibition of gamma-polymerase, but through other mechanisms associated with this has similar side effects [12].

Despite considerable research efforts related to a HIV treatment there are not known all the mechanisms of action of these drugs. The possibility of in vivo imaging of drug interaction in an organism can help to elucidate these mechanisms [13]. For the labelling of target molecules green fluorescent protein (GFP), which thanks to its fluorescent properties allows in vivo monitoring of events in a real-time, can be used [14]. The aim of this study was to assess the bond between AZT and GFP electrochemically, and further analysis of the GFP fluorescent properties influenced by the binding of AZT.

Práce je spojená s projektem NanoBioMetalNet CZ.1.07/2.4.00/31.0023

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