The effect of arsenic, cadmium, mercury, and lead on the genotoxic activity of Boletaceae family mushrooms present in Serbia.

The aim of this study was to determine accumulation of heavy metals and metalloids which are widely distributed in the environment and in food chain using wild edible mushrooms belonging to the Boletaceae family mushrooms. In addition, methanol extracts of mushrooms were tested for in vitro protective effect by the cytochalasin-B blocked micronucleus (CBMN) assay using chromosome aberrations in human peripheral lymphocytes as a model. The genotoxic activity of methanol extracts prepared at 4 different concentrations (1, 2, 3 or 6 µg/ml) was examined using amifostine and mitomycin C as positive controls. Extracts of species B. regius and B. edulis exhibited the greatest reduction in the frequency of micronuclei (MN). Extract of B. regius at concentrations of 2 µg/ml showed the highest decrease in number of MN. In comparison, extract of mushroom B. edulis at a concentration of 3 µg/ml displayed less reduction. However, as heavy metals and metalloids are found in mushrooms, another aim was to examine whether these agents affected genotoxicity. Principal component analysis (PCA) identified clustering differences between control and heavy metals and metalloids groups and might explain the influence of heavy element content and genotoxic activity in mushrooms.

Secondary metabolites of Seseli rigidum: Chemical composition plus antioxidant, antimicrobial and cholinesterase inhibition activity.

Extracts of different polarity obtained from various plant parts (root, leaf, flower and fruit) of Seseli rigidum were studied by different antioxidant assays: DPPH and ABTS radical scavenging activity, by total reducing power method as well as via total content of flavonoids and polyphenols. Essential oils of all plant parts showed weak antioxidant characteristics. The inhibitory concentration range of the tested extracts, against bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, and fungi Candida albicans and Aspergillus niger was 0.01-1.50 mg/mL and of a microbicidal 0.02-3.00 mg/mL. In the interaction with cholinesterase, all essential oils proved effective as inhibitors. The highest percentage of inhibition versus human and horse cholinesterase was shown by root essential oil (38.20% and 48.30%, respectively) among oils, and root hexane extract (40.56% and 50.65% respectively). Essential oils and volatile components of all plant parts were identified by GC, GC-MS and headspace/GC-MS. Statistical analysis of the ensemble of results showed that the root essential oil composition differed significantly from essential oils of other parts of the plant. Taking into account all of the studied activities, the root hexane extract showed the best overall properties. By means of high performance liquid chromatography coupled to high resolution mass spectrometry, the 30 most abundant constituents were identified in extracts of different polarity. The presence of identified constituents was linked to observed specific biological activities, thus designating compounds potentially responsible for each exhibited activity.

Wild fire impact on copper, zinc, lead and cadmium distribution in soil and relation with abundance in selected plants of Lamiaceae family from Vidlic Mountain (Serbia).

Fire has been considered as an improving factor in soil quality, but only if it is controlled. Severe wild fire occurred in the summer 2007 on the Vidlic Mountain (Serbia) overspreading a huge area of meadows and forests. Main soil characteristics and content of heavy metals (Cu, Pb, Cd, Zn) in different fractions obtained after sequential extraction of soil from post-fire areas and from fire non disturbed areas were studied. In four plant species of Lamiaceae family (Ajuga genevensis L., Lamium galeobdolon (L.) L., Teucrium chamaedrys L., Acinos alpinus (L.) Moench.), that grow in typical habitats of the mountain, distribution of heavy metals in aerial parts and roots was investigated too. For all samples from post-fire area cation exchange capacity and soil organic matter content are increased while rH is decreased. Fire caused slightly increased bioavailability of the observed metals but more significant rise happened in metal amounts bound to oxides and organics. The plants showed variable behavior. T. chamaedrys collected on the post-fire area contained elevated concentrations of all analyzed metals. A. alpinus showed higher phytoaccumulation for Zn and Cd, while the other two plant species for Pb and Cd in the post-fire areas.

A modification of the kinetic determination of pancuronium bromide based on its inhibitory effect on cholinesterase.

A modification of the existing spectrophotometric kinetic method for the determination of pancuronium bromide (PCBr), based on pooled human serum cholinesterase (ChE, EC 3.1.1.8 acylcholine acylhydrolase) inhibition, was developed. Butyrylthiocholine iodide (concentration 1.667 mmol/L) was used as substrate and determination was performed at pH 7.6. Essential basic kinetic parameters were also determined: Michaelis-Menten's constant KM=0.33 mmol/L, maximal reaction rate Vmax=42.29 micromol/L min, inhibition constant KI=0.34 micromol/L, and IC50=0.235 micromol/L. Linear dependence between the reaction rate and the inhibitor concentration exists in PCBr concentration range 8.29-265.28 nmol/L, which corresponds to the real sample concentrations from 0.166 to 5.306 micromol/L. The method detection limit was established to be 1.86 nmol/L and the quantification limit was 6.18 nmol/L. Precision of the method was tested for three pancuronium concentrations (16.58, 99.48, and 198.96 nmol/L). The relative standard deviation (RSD) was in the range 0.78-5.13%. Accuracy was examined by the standard addition method. The influence of substances usually present in serum and urine on the reaction rate was determined. The method developed was applied for PCBr determination in spiked serum and urine samples and in the urine taken during surgery. The method was proven to have good sensitivity, accuracy, and precision and can be considered suitable for clinical practice.

Kinetic-photometric determination of iodide based on its inhibitory effect on the bromate oxidation of Victoria Blue 4-R.

A new sensitive kinetic method has been developed for the determination of trace amounts of I- over the range of 50-300 ng cm(-3). The method was based on the inhibitory effect of iodide on the oxidation of Victoria Blue 4-R by KBrO3. The reaction is monitored spectrophotometrically at 596.3 nm. Method development includes optimization of reagent concentration and temperature. The kinetic parameters of the reaction were reported and a rate equation was suggested. The effects of certain foreign ions upon the reaction rate were determined for the assessment of the selectivity of the method. The proposed method was successfully applied to the determination of iodide in real samples. The new developed method was found to have fairly good selectivity, sensitivity, simplicity and rapidity.