Effects of bee venom and melittin on cell response of tumor and non-tumor cells in vitro

U ovom radu je istražen potencijalni protutumorski učinak pčelinjega otrova i njegove glavne sastavnice melitina prema različitim tipovima tumorskih stanica i njihovim sublinijama otpornima na citostatike kao i njihov učinak prema normalnim ne-tumorskim stanicama. Nakon obrade pčelinjim otrovom i melitinom sve stanine linije pokazale su citotoksini učinak koji je bio ovisan o dozi. Nadalje, obrada pčelinjim otrovom i melitinom uzrokovala je značajne morfološke promjene i staninu smrt uzrokovanu dominantno nekrozom. Nakon zajedničke obrade sa kemoterapeutikom cisplatinom, pčelinji otrov je pokazao aditivan učinak koji bi mogao biti koristan u vidu smanjivanja doze cisplatine za vrijeme kemoterapije. Također, pčelinji otrov i melitin uzrokovali su citotoksičnost i kod normalnih stanica uzrokujući smanjenje stanične vijabilnosti, morfološke promjene i oštećenja molekule DNA. Smanjenje glutationa i porast koncentracije malondialdehida, uz oksidativna oštećenja molekule DNA i porast aktivnosti fosfolipaze C, ukazuju da pčelinji otrov i melitin djeluju putem lipidne peroksidacije te da je oksidativni stres barem djelomično uključen u mehanizam djelovanja ovih spojeva. Iako toksičan kod svih ispitanih staninčih tipova, toksičnost pčelinjega otrova i melitina ovisna je o tipu stanica čineći tumorske stanice osjetljivijima na oba spoja naspram ne-tumorskih stanica. Ovisno o podrijetlu, stanice otporne na citostatike mogle bi biti osjetljivije na pčelinji otrov i melitin od svojih roditeljskih linija. Iako su se pokazali toksičnim kod svih ispitanih stanica, ovi rezultati idu u prilog upotrebe pčelinjega otrova i melitina u terapiji protiv tumora.Possible anticancer ability of bee venom and its major component melittin towards different types of tumour cells and their drug resistant sublines as well as their impact on normal non-tumour cells was investigated. After the treatment with whole bee venom or melittin all the cell lines tested displayed dose dependent cytotoxicity. In addition, treatment with bee venom and melittin caused significant morphological changes and induced dominantly necrotic type of cell death. Following a combined treatment with chemotherapeutic drug cisplatin, bee venom exhibited additive effect what could be useful from the point of minimizing cisplatin concentration during chemotherapy. Bee venom and melittin were toxic to normal cells as well; causing lover cell viability, morphological cell alterations and DNA damage. Depletion of glutathione and increment in malondialdehid concentration parallel with oxidative DNA damage and induction of phospholipase C activity, suggest that both bee venom and melittin act through lipid peroxidation and that oxidative stress is at least partly involved in their mode of action. Although toxic to all cell types tested, toxicity of bee venom and melittin was cell type dependent making tumour cells more sensitive to both, as compared to the non-tumour cells. Depending on the origin, drug resistant cells could be more sensitive to bee venom and melittin than parental cells. Although toxic to all the cell lines tested, our results speak in favour of application of bee venom and melittin in anticancer therapy

Efficacy of HUMN criteria for scoring the micronucleus assay in human lymphocytes exposed to a low concentration of p, p, -DDT

The cytokinesis-block micronucleus (CBMN) assay is one of the standard cytogenetic tools employed to assess chromosomal damage subsequent to exposure to genotoxic/cytotoxic agents, and is widely applicable to plant, animal and human cells. In the present study, the CBMN assay was used to assess the baseline damage in binuclear human peripheral blood lymphocytes exposed to 25 microg/L p, p'-DDT for 1, 2, 24, and 48 h by measuring the frequency of micronuclei, nucleoplasmic bridges and nuclear buds. These new scoring criteria facilitated the detection of different types of clastogenic and aneugenic effects induced by this type of pollutant. With these criteria, CBMN can also be used to measure nucleoplasmic bridges which are considered to be consequences of chromosome rearrangements and nuclear buds which are biomarkers of altered gene amplification and gene dosage. The total number of micronuclei observed in binuclear human peripheral blood lymphocytes of the exposed samples (ranging from 32 to 47) was significantly greater (P < 0.05) than that detected in the unexposed (0 time) control sample, where the total number of micronuclei was 7. The number of nucleoplasmic bridges and nuclear buds obtained after 24 and 48 h was also significantly (P < 0.05) greater in the samples treated with p, p'-DDT than in the unexposed control samples. Thus, our results confirmed the usefulness of the new criteria applicable for the CBMN assay employed in measuring the DNA damage and its role of a sensitive cytogenetic biomarker

Atorvastatin therapy safety from the aspect of genotoxicity

Primjena statina danas predstavlja jedan od osnovnih pristupa u liječenju bolesti koje su vezane uz povišenu razinu kolesterola u krvi. Osnovni cilj djelovanja ove skupine lijekova usmjeren je prema snižavanju ukupne koncentracije kolesterola u krvi inhibicijom djelovanja 3-hidroksi-3-metilglutaril koenzim A (HMG-CoA) reduktaze. Da bi se postigao što bolji terapijski učinak u što kraćem vremenskom razdoblju, primjenjuju se različite koncentracije ovih lijekova u rasponu od 10 mg/dan do 80 mg/dan. Rezultati dosadašnjih epidemioloških istraživanja pokazali su da određene koncentracije ovih lijekova mogu ostaviti posljedice na okolne stanice i tkiva nakon dulje izloženosti, od kojih su najčešće bolesti skeletnih mišića, jetrene bolesti, povećane vrijednosti serumskih transaminaza, te miopatija koja može voditi u rabdomiolizu i bubrežno zatajenje. Istraživanja lijekova koji se koriste za snižavanje razine kolesterola u krvi provode se od njihovog otkrića u ranim sedamdesetim godinama prošloga stoljeća. Iako su se pristupi istraživanjima mijenjali tijekom godina, cilj im je uvijek bio isti – pronaći one najpogodnije te one doze u kojima bi njihova djelotvornost bila najučinkovitija, a rizik od mogućih posljedica najmanji. Ovaj rad stoga donosi pregled dosadašnjih istraživanja vezanih uz sigurnost atorvastatina u terapijske svrhe i smjernice za provođenje protokola za tesiranje lijekova.The application of statin today represents one of the basic approaches to the treatment of the disease connected to high levels of cholesterol in the blood. Statins are a widely used group today found in different generic names of cholesterol-lowering agents that act by inhibiting 3-hydroxy 3-methylglutaryl CoA (HMG CoA) reductase, an enzyme which catalyses the rate-limiting step in cholesterol biosynthesis. In order to achieve the best therapeutical effects in the shortest period of time various concentrations of these drugs are administered in the range of containing 10 mg/day to 80 mg/day atorvastatin. The results of recent epidemiological researches have shown that determined concentrations of these drugs administered over a longer period can have serious side effects. The most important adverse effects in clinical practice are disease of the skeletal muscles, asymptomatic increases in liver transaminases, progressive liver disease and myopathy which can lead to rhabdomyolysis and cause acute renal failure. Researches regarding this group of drugs have been conducted since the early seventies of the last century with the same effort; to find suitable dosage that would be effective in reducing the cholesterol level. Even though the research approaches have changed through the years, the aim has always been the same; to find the most adequate and most effective dosage with the lowest side effect risk. This review paper brings therefore an overview of recent researches regarding atorvastatin therapy safety with guidelines for protocols used in drug research

Presence of Dichlorodiphenyltrichloroethane (DDT) in Croatia and Evaluation of Its Genotoxicity

The DDT was one of the most commonly used pesticide in mid 20th century and even though its use is banned it is detectible in water, soil, fish, meat and milk products. Using comet assay and micronucleus test we have managed to detect genotoxic properties of this pesticide on human peripheral blood lymphocytes. Results obtained in this research indicate the need for further environmental and food monitoring, and cytogenetic research using sensitive methods in detection of primary genome damage after exposure to DDT to establish the impact of such chemicals on human genome and health

Working Group “Yellow”. Environmental pollution monitoring and adverse effects of chemicals used in food production (FoCUS – Food Chemicals Used Safely)

This project addresses the need to assess the environmental and health risks of chemicals used in food production, since their release into the environment may lead to different ecological effects. The occurrence and effects of selected chemicals on wildlife and humans will be addressed to provide data sets, which are necessary for scientifically-based risk assessment. Special emphasis will be put on the combined effects of environmentally relevant mixtures. A combination of state-of-the-art methods will be applied to predict synergistic and/or additive effects of combined exposure. Development and implementation of new technologies for waste treatment and re-use of food industry by-products by converting them into value-added items will also be a significant task

Workgroup A. BioBricks. The Fate and Effetct of Sewage Sludge-Based Bricks on Human Health and Water Resource Quality

The overall project goal is to prevent water pollution in the River Basin of the Danube’s stretch crossing Croatia, Serbia and Hungary, coming from the use of sewage sludge-based bricks in house construction, which despite representing a solution to the ever increasing problem of waste management and determining a reduction of the clay needed for construction, may pose environmental and health dangers if not properly used, due to leaching of toxic substances and heavy metals into the waters. The project aims at creating a comprehensive database on SSBB environmental and health effects; developing a guideline for SSBB manufacturers on standardised SSBB production, and increasing the awareness and knowledge of manufacturers and consumers on possible negative effects and the proper handling/use of SSBBs