Chemical Composition and Protective Possibilities of Juglans Nigra Leaves and Green Husks Extracts: DNA Binding and Micronucleus Assay in Human Lymphocytes.

To better understand the mechanism of action of the compounds in the ethanolic extracts of J. nigra leaves and green husks, their binding to CT-DNA was investigated. This study was conducted to elucidate the in vitro protective effect of extracts against chromosomal damage in mitogen-induced human lymphocytes and investigate the possible application of selec+ted extracts as a natural source of polyphenolic compounds. Using HPLC-MS analysis, 103 different compounds were identified as having a higher number of active species, which is consistent with their activity. The frequency of micronuclei (MN) was scored in binucleated cells, and the nuclear proliferation index was calculated. Cyclic voltammetry experiments demonstrate that the nature of the interaction between extracts and CT-DNA is a synergy of electrostatic and intercalative modes, where leaves extracts showed a higher ability to bind to DNA. Extracts showed excellent antioxidant activity. At a concentration of only 4 µg/mL, extract of J. nigra leaves and the green husks reduced the incidence of MN by 58.2% and 64.5%, respectively, compared to control cell cultures.

Optimization of radioprotective dose of Juglans nigra leaf extract using response surface methodology.

J. nigra leaf contains mixture of various pharmacologically active compounds and it is assumed that they may have the potential radioprotective effect. The purpose of this work was to predict radioprotective doses by correlating changes in organ distribution of radiopharmaceuticals with extract dose levels and rat body weight using response surface methodology (RSM) based on a second-order polynomial equation. The extract was administered daily by oral gavage to rats at dose of 6.9, 10.3, or 13.7 mg kg-1 body weight (bw) day-1 for 10 days. On the eleventh day, 0.1 ml of the one radiopharmaceutical (Na99mTcO4, 99mTc-dimercaptosuccinic acid and 99mTc-tin colloid) was injected into the tail vein. The statistical parameters: the coefficient of determination (0.81-0.95), the coefficient of variation (3.05-11.1), the adequate precision (8.82-19.12) and the mean relative percentage deviation (± 2.3-8.2) were indicated the precision and reliability of RSM. Using RSM, the predicted daily dose of leaf extract ranging from 11.19 to 11.99 mg kg-1 bw may be considered as an optimal daily radiopotective dose for protection of organs from radiation in cases of planned radiation exposures.

Modelling and optimisation of treatment parameters in high-dose-rate mono brachytherapy for localised prostate carcinoma using a multilayer artificial neural network and a genetic algorithm: Pilot study.

BACKGROUND: High-dose-rate mono brachytherapy (HDR-MB) is employed in the treatment of prostate carcinoma (CaP). As an ideal plan of CaP brachytherapy cannot be created, it is necessary to identify a reliable tool to optimise the parameters of HDR-MB. This paper applies a multilayer artificial neural network (MANN) and a genetic algorithm (GA) to optimise brachytherapy parameters based on an individual dose-volumetric analysis. METHODS: Patients with localised CaP of various risks were treated with HDR-MB. Consecutive levels of the biochemical control parameter (prostate specific antigen (PSA) nadir) have been collected after completion of HDR-MB in the range 2-9 years. The Kaplan-Meier regression analysis of biochemical-free survival (BFS) was applied. The clinical risk of recurrent CaP (RCaP), the therapy dose (TD), TD coverage index (CI100%) and PSA nadir were modelled using the MANN and GA. RESULTS: In the low-risk group, BFS was achieved in 100% of treated patients, while in the group of patients with high risk, BFS was achieved in 95.8% of treated patients. The MANN-GA model optimises a TD of 47.3 Gy and CI100% of 1.14 as well as a TD of 50.4 Gy and CI100% of 1.6 for the low-risk group and high-risk group, respectively, of localised CaP. The optimised PSA nadir was 0.047 and 0.25 ng cm-3 for low-risk group and high-risk group, respectively. CONCLUSIONS: The developed MANN-GA model presents a method for optimising the treatment parameters in radiation therapy, which could be a valuable tool in planning of the HDR-MB.

In vitro protease inhibition and cytotoxicity of Aspergillus fumigatus biomolecules secreted under long-term aerated conditions.

The fatality rate of invasive aspergillosis (IA) is still very high, especially in prolonged and untreated pulmonary cases. Aspergillus fumigatus is the main causative agent of IA and investigation of its metabolites could provide valuable insight into virulence factor(s) associated with this organism. We evaluated the A. fumigatus culture filtrate (CF) products generated during short- and long-term aerated and non-aerated conditions and tested for (i) inhibition of cysteine or serine proteases and (ii) cytotoxicity. In addition, the mathematical model was determined using response surface methodology (RSM) to estimate the influence of different fermentation conditions on A. fumigatus CF characteristics, predict enzyme inhibition and make possible correlations with in vivo conditions. Biosynthesis of A. fumigatus low molecular weight proteinaceous products (from 6.4 to 15.4 kDa) was observed after 6 days of growth under aerated and alkaline conditions. Also, only these CFs showed significant reduction in cell lines survival (Caco-2 and WISH 35.6% and 54.6%, respectively). Obtained results provide solid starting point for further studies that would include: (i) detailed chemical characterization of A. fumigatus CF, (ii) activity relationships and in vivo correlation with pathogenicity of prolonged pulmonary IA and (iii) possible use of biomolecules as diagnostic or therapeutic markers.