No abstract

We examined the influence of caffeine on honeybee lifespan, Nosema resistance, key enzyme activities, metabolic compound concentrations, and total DNA methylation levels. Caffeine slowed age-related metabolic tendencies. Bees that consumed caffeine lived longer and were not infested with Nosema spp. Caffeine-treated workers had higher protein concentrations. The levels increased with aging but they then decreased in older bees. Caffeine increased the activities of antioxidant enzymes (SOD, GPx, CAT, GST), AST, ALT, ALP, neutral proteases, and protease inhibitors, and the concentrations of uric acid, triglycerides, cholesterol, glucose, and Ca2+. Acidic and alkaline protease activities were lower in the bees treated with caffeine. Creatinine and Mg2+ concentrations were higher in the caffeine-treated workers but only up to 14 days of age. Caffeine significantly decreased DNA methylation levels in older bees. The compound could be considered as a natural diet supplement increasing apian resistance to stress factors. Our studies will enhance possibilities of using Apis mellifera as a model organism in gerontological studies.

Three bioactive fractions, extracellular laccase (ex-LAC), crude endopolysaccharides (c-EPL), and a low molecular subfraction of secondary metabolites (ex-LMS), were isolated from the idiophasic cultures of the white rot fungus Cerrena unicolor. For the first time, we determined the antioxidant properties of these samples by chemiluminometric measurement (a) and assessment of the scavenging effect on ABTS (b) and the DPPH reduction rate (c). The highest reducing capability was found for the ex-LMS fraction: 39–90% for (a), 20–90% for (b), and 10–59% for (c) at the concentration of 6.25–800 µg/mL. The scavenging abilities of the C. unicolor c-EPL were between 36 and 70% for (a), 2 and 60% for (b), and 28 and 32% for (c) at the concentration of 6.25–800 µg/mL. A very high prooxidative potential was observed for the ex-LAC probes. The preliminary toxicity tests were done using the Microtox system and revealed the following percentage of the toxic effect against Vibrio fischeri: 85.37% for c-EPL, 50.67% for ex-LAC, and 99.8% for ex-LMS, respectively. The ex-LAC sample showed the antibacterial activity against Escherichia coli, c-EPL against Staphylococcus aureus, and ex-LMS against both bacterial strains, respectively, but the stronger inhibitory effect was exerted on S. aureus.

No abstract

No abstract

-We describe how protease and protease inhibitor activity patterns vary during ontogenesis, with season, and in relation to caste and sex in the honey bee (Apis mellifera). Extraction of body surfaces with water and detergent was followed by the in vitro analysis of proteolytic activity and protease inhibitor level, as well as the electrophoretic separation of extracts in polyacrylamide gels. In in vitro experiments, we compared two groups of detectable proteolytic activities: neutral and alkaline water-soluble versus acidic detergent-soluble. The most active proteases appeared to be acidic ones and were detected on drone pupae in spring. The most distinct and most active protease bands in electrophoretic separations were those obtained for neutral and alkaline activities on queens in all seasons. The highest levels of protease inhibitor activities in vitro were obtained from worker samples in all seasons. The enzymatic properties suggest that all catalytic types of proteases were present in the extracts, but at different activity levels, depending on pH: asparagine and cysteine proteases at pH 2.5, cysteine proteases and metalloproteases at pH 7.0, and serine proteases at pH 11.5, respectively. honey bee / proteases / protease inhibitors / body surface proteolysis / zymography

It is known that earthworm coelomic fluid (CF) can affect not only cancer but also normal cells. The study demonstrated that the CF of the earthworm Dendrobaena veneta exhibited cytotoxicity against A549 lung cancer cells but did not toward the bronchial epithelial cell line BEAS‐2B. The selective effect on the tumor cells was achieved after a short‐term CF heat pre‐treatment at 70 °C. The cytotoxic effect of the CF was time‐ and concentration‐dependent. The CF noticeably decreased the viability and affected the morphology of the A549 cells. Scanning electron microscopy revealed a different degree of destruction of the nucleus and cytoplasm of A549 cells. As determined by atomic force microscopy, the cell surface roughness increased while the cell stiffness was reduced upon the CF treatment. A twofold increase in the caspase 3, 4, 5, and 10 levels was observed in the A549 cells after the incubation with the CF. The results obtained by flow cytometry using Annexin V confirmed the proapoptotic effect of the earthworm CF on A549 lung cancer cells. The D. veneta CF and active fraction obtained with cytotoxicity toward A549 lung cancer is an interesting and promising preparation for further biological, chemical, and biomedical research.

Aims: The aim of our research was to isolate the compounds from the metabolites of Raoultella ornithinolytica with the activity against Candida albicans and to analyse the action of the compounds on the metabolic activity and morphology of the fungus cells. Methods and Results: The effect of active protein fractions on the cell morphology, growth, and metabolic activity of C. albicans was analysed under a light microscope with Nomarski contrast and after staining with calcofluorwhite. The LIVE/DEAD Yeast Viability Kit F-7030 FUN 1 was used for determination of C. albicans metabolic activity. The biomolecules obtained after isolation by ion exchange chromatography were further fractionated by Sephadex G-50 medium gel filtration. Then, after molecular sieve, the fractions were analysed by FTIR and SERS Spectroscopy. A subfraction was isolated from the antifungal protein fraction above 100 kDa. The active subfraction identified as the glyco-protein complex caused a decrease in the metabolic activity and morphological changes of C. albicans cells. Conclusions: The glyco-protein complex obtained from metabolites of bacteria Raoultella ornithinolytica possesses antifungal activity against C. albicans and shows minimal toxicity (1%) against fibroblasts. Significance and Impact of the Study: Studies on the glyco-protein complex obtained from earthworm gut bacteria R. ornithinolytica can lead to their application in biological fungicide and pharmaceutical industry.