Ocjena apsorpcije, djelotvornosti protiv bakterije Escherichia coli i citotoksičnosti krutih lipidnih nanočestica s moksifloksacinom

Moxifloxacin (MOX) is an important antibiotic commonly used in the treatment of recurrent Escherichia coli (E. coli) infections. The aim of this study was to investigate its antibacterial efficiency when used with solid lipid nanoparticles (SNLs) and nanostructured lipid carriers (NLCs) as delivery vehicles. For this purpose we designed two SLNs (SLN1 and SLN2) and two NLCs (NLC1 and NLC2) of different characteristics (particle size, size distribution, zeta potential, and encapsulation efficiency) and loaded them with MOX to determine its release, antibacterial activity against E. coli, and their cytotoxicity to the RAW 264.7 monocyte/macrophage-like cell line in vitro. With bacterial uptake of 57.29 %, SLN1 turned out to be significantly more effective than MOX given as standard solution, whereas SLN2, NLC1, and NLC2 formulations with respective bacterial uptakes of 50.74 %, 39.26 %, and 32.79 %, showed similar activity to standard MOX. Cytotoxicity testing did not reveal significant toxicity of nanoparticles, whether MOX-free or MOX-loaded, against RAW 264.7 cells. Our findings may show the way for a development of effective lipid carriers that reduce side effects and increase antibacterial treatment efficacy in view of the growing antibiotic resistance.Moksifloksacin je važan antibiotik koji se često rabi za liječenje rekurentne infekcije bakterijom Escherichia coli (E. coli). Cilj je ovog istraživanja bio ocijeniti njegovu djelotvornost u formulaciji s krutim lipidnim nanočesticama (engl. solid lipid nanoparticles, krat. SNL) i nanostrukturiranim lipidnim nosačima (engl. nanostructured lipid carriers, krat. NLC) kao njegovim vehikulima. U tu smo svrhu osmislili dva SLN-a (SLN1 I SLN2) te dva NLC-a (NLC1 i NLC2) različitih svojstava (veličine čestice, raspodjele veličina, zeta potencijala i sposobnosti enkapsulacije) te ih obogatili moksifloksacinom kako bismo utvrdili njegovo otpuštanje, djelovanje protiv E. coli i citotoksičnost za makrofagnu staničnu liniju RAW 264.7 in vitro. S bakterijskom apsorpcijom od 57,29 %, SLN1 se pokazao značajno djelotvornijim vehikulom moksifloksacina od njegove standardne formulacije (otopine), a formulacije s SLN2, NLC1 odnosno NLC2 s odgovarajućim apsorpcijama od 50,74 %, 39,26 % odnosno 32,79 % iskazale su djelotvornost sličnu onoj standardnog antibiotika. Test citotoksičnosti nije pokazao značajnu toksičnost nanočestica bez obzira na to jesu li sadržavale moksifloksacin ili nisu. Naši rezultati upućuju na mogući smjer razvoja djelotvornih lipidnih nosača kojima bi se mogle smanjiti nuspojave i povećati antibakterijska djelotvornost liječenja s obzirom na sve veću bakterijsku rezistentnost

Genotoxicity evaluation of naringin on mammalian cell lines by micronucleus assay

53rd Congress of the European-Societies-of-Toxicology (EUROTOX) -- SEP 10-13, 2017 -- Bratislava, SLOVAKIAWOS: 000425486700267…European Soc Toxico

DNA damage assessment with buccal micronucleus cytome assay in Turkish coal miners

The aim of this study was to assess DNA damage in Turkish coal miners with the buccal micronucleus cytome (BMCyt assay as the least invasive and therefore most practical method that may find wider application in coal miner biomonitoring. Buccal epithelial cell samples were taken from 54 coal miners and 42 controls from Zonguldak, Turkey to establish their micronucleus (MN), binucleus (BN), condensed chromatin (CC), karyorrhectic (KHC), karyolytic (KYL), nuclear bud (NBUD), and pyknotic (PYC) frequencies. We also analysed the effects of confounding factors such as age, years of work at the mine, smoking, alcohol drinking, and use of protective equipment on differences in MN frequencies. Two miners had confirmed and three suspect pneumoconiosis, whereas 49 displayed normal chest radiographs. MN, BN, KHC, and NBUD frequencies were significantly higher in coal miners than controls. Years of work at the mine also showed a significant effect on buccal MN frequencies in coal miners, but we found no correlation between MN frequencies and age, smoking, and alcohol consumption. In conclusion, BMCyt assay proved itself an accurate and practical screening method, as it can detect DNA damage much earlier than pneumoconiosis develops.Cilj je ovoga istraživanja bio procijeniti oštećenje DNA u turskih rudara primjenom tzv. Cytome inačice mikronukleustesta na bukalnim epitelnim stanicama (engl. buccal micronucleus cytome assay, krat. BMCyt) kao najmanje invazivne i stoga najpraktičnije metode koja bi mogla naći svoju primjenu u biomonitoringu rudara u ugljenokopima. Uzorci bukalnih epitelnih stanica prikupljeni su od 54 rudara i 42 kontrolna ispitanika iz Zonguldaka u Turskoj radi utvrđivanja učestalosti mikronukleusa (MN), binuklearnih stanica (BN), zgusnutoga kromatina (CC), karioreksije (KHC), kariolize (KYL), jezgrinih pupova (NB) i piknoze (PYC). Osim toga, analizirali smo koliko čimbenici poput dobi, godina staža u rudniku, pušenja, pijenja alkohola i uporabe zaštitne opreme utječu na razlike u nalazima učestalosti MN-a. Dva su rudara imala potvrđenu dijagnozu pneumokonioze, u tri je rudara postojala sumnja na nju, a ostalih 49 rudara imalo je uredne rendgenske nalaze. Učestalosti MN-a, BN-a, KHC-a i NB-a bile su značajno više u rudara nego u kontrolnih ispitanika. Godine staža u rudniku također su značajno pridonijele povišenoj učestalosti MN-a u rudara, ali takva korelacija nije utvrđena za dob, pušenje i pijenje alkohola. BMCyt test pokazao se preciznim i praktičnim probirnim testom jer otkriva oštećenje DNA mnogo ranije nego što se pojavi pneumokonioza

Procjena oštećenja DNA bukalnih epitelnih stanica primjenom Cytome inačice mikronukleus-testa

The aim of this study was to assess DNA damage in Turkish coal miners with the buccal micronucleus cytome (BMCyt assay as the least invasive and therefore most practical method that may find wider application in coal miner biomonitoring. Buccal epithelial cell samples were taken from 54 coal miners and 42 controls from Zonguldak, Turkey to establish their micronucleus (MN), binucleus (BN), condensed chromatin (CC), karyorrhectic (KHC), karyolytic (KYL), nuclear bud (NBUD), and pyknotic (PYC) frequencies. We also analysed the effects of confounding factors such as age, years of work at the mine, smoking, alcohol drinking, and use of protective equipment on differences in MN frequencies. Two miners had confirmed and three suspect pneumoconiosis, whereas 49 displayed normal chest radiographs. MN, BN, KHC, and NBUD frequencies were significantly higher in coal miners than controls. Years of work at the mine also showed a significant effect on buccal MN frequencies in coal miners, but we found no correlation between MN frequencies and age, smoking, and alcohol consumption. In conclusion, BMCyt assay proved itself an accurate and practical screening method, as it can detect DNA damage much earlier than pneumoconiosis develops.Cilj je ovoga istraživanja bio procijeniti oštećenje DNA u turskih rudara primjenom tzv. Cytome inačice mikronukleustesta na bukalnim epitelnim stanicama (engl. buccal micronucleus cytome assay, krat. BMCyt) kao najmanje invazivne i stoga najpraktičnije metode koja bi mogla naći svoju primjenu u biomonitoringu rudara u ugljenokopima. Uzorci bukalnih epitelnih stanica prikupljeni su od 54 rudara i 42 kontrolna ispitanika iz Zonguldaka u Turskoj radi utvrđivanja učestalosti mikronukleusa (MN), binuklearnih stanica (BN), zgusnutoga kromatina (CC), karioreksije (KHC), kariolize (KYL), jezgrinih pupova (NB) i piknoze (PYC). Osim toga, analizirali smo koliko čimbenici poput dobi, godina staža u rudniku, pušenja, pijenja alkohola i uporabe zaštitne opreme utječu na razlike u nalazima učestalosti MN-a. Dva su rudara imala potvrđenu dijagnozu pneumokonioze, u tri je rudara postojala sumnja na nju, a ostalih 49 rudara imalo je uredne rendgenske nalaze. Učestalosti MN-a, BN-a, KHC-a i NB-a bile su značajno više u rudara nego u kontrolnih ispitanika. Godine staža u rudniku također su značajno pridonijele povišenoj učestalosti MN-a u rudara, ali takva korelacija nije utvrđena za dob, pušenje i pijenje alkohola. BMCyt test pokazao se preciznim i praktičnim probirnim testom jer otkriva oštećenje DNA mnogo ranije nego što se pojavi pneumokonioza

EFFECTS OF EXPOSURE TO HAZARDOUS SUBSTANCES IN POTTERY AND CERAMIC WORKERS

During the last decade, the production and hence, the amount and the types of exposure of hazardous chemicals have been increased in the occupationally exposed workers. Most of these chemicals have deterious effects in the living systems. The level of chemical exposure in the occupational settings and the biomonitoring of workers and also establishing the regulatory endpoints are very important. The exposures of chemicals in the workplaces have been associated with the increase in allergy, organ and system disorders and cancer. Therefore, the removal of these hazardous substances or the substitutions of them with non-toxic ones are necessary for the protection of workers’ health. In a large variety of occupations such as pottery and ceramic industry, workers may be exposed to the many chemicals mainly to crystalline silica. Occupational exposure to crystalline silica dust is related to an increased risk for pulmonary diseases such as silicosis, tuberculosis, chronic bronchitis and lung cancer. The genotoxic changes as assessed by comet assay in foundry and ceramic workers will be given. In the presentation, the effects of age, smoking, alcohol and protective equipment usage on the genotoxicity parameters will also be discussed

Resveratrol Protects Sepsis-Induced Oxidative DNA Damage in Liver and Kidney of Rats

Background: The increases of free radicals have been proposed to be involved in the pathogenesis of sepsis, which leads to multiple-organ dysfunction syndromes. The uses of antioxidants as a complementary tool in the medical care of oxidative stress-related diseases have attracted attention of researchers. Resveratrol (RV) has suggested being antioxidant, anti-proliferative, and anti-inflammatory effects in various experimental models and clinical settings. Aims: This study was undertaken to evaluate the protective effects of RV on oxidative DNA damage induced by sepsis in the liver and kidney tissues of Wistar albino rats. Study Design: Animal experimentation. Methods: Four experimental groups consisting of eight animals for each was created using a total of thirty-two male Wistar albino rats. Sham group was given 0.5 mL of saline intra-peritoneal (ip) only following laparatomy. Sepsis group was given 0.5 mL saline ip only following the induction of sepsis. RV-treated group was given a dose of 100 mg/kg ip RV in 0.5 mL saline following laparatomy. RV-treated sepsis group was given 100 mg/kg ip RV in 0.5 mL saline following the induction of sepsis. A model of sepsis was created by cecal ligation and puncture technique. In the liver and kidney tissues, oxidative stress parameters (malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPX)) and a proinflammatory cytokine (tumor necrosis factor alpha (TNF-alpha)), were evaluated spectrophotometrically and DNA damage was determined by the alkaline single cell gel electrophoresis (comet assay) technique using formamidopyrimidine DNA glycosylase protein. Results: In the RV-treated sepsis group, the levels of MDA and TNF-alpha were lower and GSH levels, SOD and GPX activities were higher than in the septic rats (p<0.05). RV treatment significantly reduced the sepsis-induced oxidative DNA damage in the liver and kidney cells (p<0.05). Conclusion: It is suggested that RV treatment might reduce the sepsis-induced oxidative DNA damages in sepsis-related diseases; however, there is a need for more studies to clear up the protective mechanisms of RV against sepsis

Can occupational exposure to crystalline silica in ceramic industries affect the immune parameters?

53rd Congress of the European-Societies-of-Toxicology (EUROTOX) -- SEP 10-13, 2017 -- Bratislava, SLOVAKIAWOS: 000425486700564…European Soc Toxico

Preventive role of Pycnogenol (R) against the hyperglycemia-induced oxidative stress and DNA damage in diabetic rats

WOS: 000458712400006PubMed: 30465898Diabetes mellitus, a complex progressive metabolic disorder, leads to some oxidative stress related complications. Pycnogenol (R) (PYC), a plant extract obtained from Pinus pinaster, has been suggested to be effective in many diseases including diabetes, cancer, inflammatory and immune system disorders. The mechanisms underlying the effects of PYC in diabetes need to be elucidated. The aim of this study was to determine the effects of PYC treatment (50 mg/kg/day, orally, for 28 days) on the DNA damage and biochemical changes in the blood, liver, and kidney tissues of experimental diabetic rats. Changes in the activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase enzymes, and the levels of 8-hydroxy-2'-deoxyguanosine, total glutathione, malondialdehyde, insulin, total bilirubin, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, high density lipoprotein, low density lipoprotein, total cholesterol, and triglyceride were evaluated. DNA damage was also determined in the whole blood cells and the liver and renal tissue cells using the alkaline comet assay. PYC treatment significantly ameliorated the oxidative stress, lipid profile, and liver function parameters as well as DNA damage in the hyperglycemic rats. The results show that PYC treatment might improve the hyperglycemia-induced biochemical and physiological changes in diabetes.Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [114S919]This work was supported by The Scientific and Technological Research Council of Turkey (Grant number: 114S919)