The title method was used for phytoremediation of soils. The experiments were performed with leaves of willow (S. smithiana, S. alba, S. rubens and S. dasyclados) grown in soil contaminated with heavy metals (Cd, Zn, Pb), The ability of the leaves to accumulate these metals was confirmed. The total contents of metals in 0.02 mol l?1 Tris buffer (pH 7.5) and in 1 mol l?1 sodium hydroxide extracts were determined. Size exclusion chromatography combined with inductively coupled plasma mass spectrometry was used for separation and determination of metals in the extracts. The metals in the extracts are present mainly in low-molecular-weight fractions (M ? 2 kDa). These fractions were obtained by preparative size exclusion chromatography and refined by immobilized metal affinity chromatography. The compounds in the fractions were analyzed by amino acid analysis and MALDI mass spectrometry. The results show that majority of heavy metals in willow biomass remained in insoluble residue whereas the extracts contain substantial amounts of chelating agents, which suggests their role in metal detoxication processes in willow plants.
- MeSH
- chromatografie afinitní metody MeSH
- financování organizované MeSH
- látky znečišťující půdu analýza MeSH
- listy rostlin chemie MeSH
- monitorování životního prostředí MeSH
- regenerace a remediace životního prostředí metody MeSH
- Salix metabolismus růst a vývoj MeSH
- stopové prvky izolace a purifikace MeSH
- těžké kovy analýza MeSH
The chemical modification of amino acid side-chains followed by mass spectrometric detection can reveal at least partial information about the 3-D structure of proteins. In this work we tested diethylpyrocarbonate, as a common histidyl modification agent, for this purpose. Appropriate conditions for the reaction and detection of modified amino acids were developed using angiotensin II as a model peptide. We studied the modification of several model proteins with a known spatial arrangement (insulin, cytochrome c, lysozyme and human serum albumin). Our results revealed that the surface accessibility of residues is a necessary, although in itself insufficient, condition for their reactivity; the microenvironment of side-chains and the dynamics of protein structure also affect the ability of residues to react. However the detection of modified residues can be taken as proof of their surface accessibility, and of direct contact with solvent molecules.
- MeSH
- diethylpyrokarbonát analýza chemie MeSH
- financování organizované MeSH
- histidin chemie MeSH
- koně MeSH
- kur domácí MeSH
- lidé MeSH
- lysin chemie MeSH
- molekulární struktura MeSH
- povrchové vlastnosti MeSH
- proteiny analýza chemie MeSH
- spektrofotometrie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
INTRODUCTION: Various dysfibrinogenemias have been described worldwide. This paper describes two new cases of dysfibrinogenemia identified in the Czech Republic. MATERIALS AND METHODS: The proposita of fibrinogen Novy Jicin, a 12-year-old girl, presented with hemorrhagic complications, low Clauss fibrinogen level (0.3 g/l) and prolonged both thrombin (70.8 s) and reptilase (>180 s) time. Her mother and sister both presented with normal coagulation tests, normal fibrinogen level and reported no history of bleeding. The carriers of the fibrinogen Praha II were a 31-year-old man and his 11-year-old daughter. They both presented with low fibrinogen Clauss level (0.88 g/l) and prolonged thrombin and reptilase time. To identify the genetic mutation responsible for these dysfibrinogens, genomic DNA extracted from the blood was analyzed. The presence of the mutant chains in the circulation was determined by MALDI-TOF mass spectroscopy. Scanning electron micrographs of the patients' fibrin clots were obtained. RESULTS: The kinetics of fibrinopeptide release and fibrin polymerization were impaired for both fibrinogen Novy Jicin and Praha II. DNA sequencing showed heterogeneous fibrinogen Aalpha R16C mutation in the fibrinogen Novy Jicin case and heterogeneous fibrinogen Aalpha R16H in the fibrinogen Praha II case. The mutant chains were found to be expressed to the circulation by MALDI-TOF mass spectroscopy. Scanning electron micrographs of the patient's fibrin clot were found to be abnormal. CONCLUSIONS: The case of dysfibrinogenemia Aalpha R16C-fibrinogen Novy Jicin and the case of dysfibrinogenemia Aalpha R16H were found by routine coagulation testing and were genetically identified.
- MeSH
- afibrinogenemie genetika MeSH
- dítě MeSH
- dospělí MeSH
- fibrin metabolismus MeSH
- fibrinogeny abnormální genetika MeSH
- kinetika MeSH
- krvácení MeSH
- lidé MeSH
- missense mutace MeSH
- mutační analýza DNA MeSH
- vyšetření krevní srážlivosti MeSH
- zdraví rodiny MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Geografické názvy
- Česká republika MeSH
- Publikační typ
- abstrakt z konference MeSH
