Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution

Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MT using an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4 °C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 μg ml(-1) of MT (50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(i) ions (50 μM AgNO(3)). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(i) ions was determined by high performance liquid chromatography with electrochemical detection.     

Charging systems for municipal solid waste: experience from the Czech Republic

The paper presents results of research into municipal waste treatment in the Czech Republic. Its special focus is on the impacts of various municipal solid waste charging systems on separating and recycling efforts of municipalities and households. The municipal solid waste charging systems are shortly described first, including the principles of the relevant Czech legislation. It shows that the Czech waste legislation provides space for implementing Pay-as-You-Throw (PAYT) models in the Czech Republic. The main results of representative surveys conducted by the authors within the EU PAYT project in 2003 in selected Czech municipalities and Prague households are shown. The survey confirmed that in municipalities that apply the PAYT charging system, citizens separate more waste and produce less residual waste. The survey data analysis has also shown which factors contributing to satisfactory waste separation are relevant and should be taken into the account when providing policy recommendations for introducing PAYT charging systems in other cities.