Red blood cells (RBCs) are the most abundant cells in the human blood that have been extensively studied under morphology, ultrastructure, biochemical and molecular functions. Therefore, RBCs are excellent cell models in the study of biologically active compounds like drugs and toxins on the structure and function of the cell membrane. The aim of the present study was to explore erythrocyte ghost’s proteome to identify changes occurring under the influence of three bee venom peptides-melittin, tertiapin, and apamin. We conducted preliminary experiments on the erythrocyte ghosts incubated with these peptides at their non-hemolytic concentrations. Such preparations were analyzed using liquid chromatography coupled with tandem mass spectrometry. It was found that when higher concentrations of melittin and apamin were used, fewer proteins were identified. Moreover, the results clearly indicated that apamin demonstrates the greatest influence on the RBCs ghosts proteome. Interestingly, the data also suggest that tertiapin exerted a stabilizing effect on the erythrocyte membrane. The experiments carried out show the great potential of proteomic research in the projects focused on the toxin’s properties as membrane active agents. However, to determine the specificity of the effect of selected bee venom peptides on the erythrocyte ghosts, further proteomic research should be focused on the quantitative analysis.
Melittin (MEL) is a basic polypeptide originally purified from honeybee venom. MEL exhibits a broad spectrum of biological activity. However, almost all studies on MEL activity have been carried out on vertebrate models or cell lines. Recently, due to cheap breeding and the possibility of extrapolating the results of the research to vertebrates, insects have been used for various bioassays and comparative physiological studies. For these reasons, it is valuable to examine the influence of melittin on insect physiology. Here, for the first time, we report the immunotropic and cardiotropic effects of melittin on the beetle Tenebrio molitor as a model insect. After melittin injection at 10−7 M and 10−3 M, the number of apoptotic cells in the haemolymph increased in a dose-dependent manner. The pro-apoptotic action of MEL was likely compensated by increasing the total number of haemocytes. However, the injection of MEL did not cause any changes in the percent of phagocytic haemocytes or in the phenoloxidase activity. In an in vitro bioassay with a semi-isolated Tenebrio heart, MEL induced a slight chronotropic-positive effect only at a higher concentration (10−4 M). Preliminary results indicated that melittin exerts pleiotropic effects on the functioning of the immune system and the endogenous contractile activity of the heart. Some of the induced responses in T. molitor resemble the reactions observed in vertebrate models. Therefore, the T. molitor beetle may be a convenient invertebrate model organism for comparative physiological studies and for the identification of new properties and mechanisms of action of melittin and related compounds.
Oviductal epithelial cells (OECs) actively produce stimulating and protecting factors, favoring survival and viability of gametes and early embryos. The oviduct participates in the initial reproductive events, which strongly depends on adhesion. The analysis of differential gene expression in OECs, during long-term in vitro culture, enables recognition of new molecular markers regulating several processes, including “biological adhesion”. Porcine oviducts were stained with hematoxylin and eosin, as well as with antibodies against epithelial markers. Then, OECs were long-term in vitro cultured and after 24 h, 7, 15, and 30 days of culture were subjected to transcriptomic and proteomic assays. Microarrays were employed to evaluate gene expression, with Matrix-assisted laser desorption/ionization-time of light (MALDI-TOF) mass spectrometry applied to determine the proteome. The results revealed proper morphology of the oviducts and typical epithelial structure of OECs during the culture. From the set of differentially expressed genes (DEGs), we have selected the 130 that encoded proteins detected by MALDI-TOF MS analysis. From this gene pool, 18 significantly enriched gene ontology biological processes (GO BP) terms were extracted. Among them we focused on genes belonging to “biological adhesion” GO BP. It is suggested that increased expression of studied genes can be attributed to the process of intensive secretion of substances that exhibit favorable influence on oviductal environment, which prime gametes adhesion and viability, fertilization, and early embryo journey.
Dietary supplements cover a wide range of products, the most popular are those containing plant-based ingredients. Supplements are consumed by consumers of all ages as well as by both healthy and sick people. The lack of unified regulation in this sector increases the probability that supplements are poor chemical and microbiological quality and can be dangerous for patients. The aim of this paper is to highlight selected issues associated with the microbiological quality of dietary supplements containing plant materials. We focus on the most recent reports referring to bacterial and fungal contaminations as well as the presence of mycotoxins. Dietary supplements containing plant ingredients commonly show a variety of microbial contaminants, which might be crucial for consumer safety. They often contain microorganisms potentially pathogenic to humans. Metabolites produced by microorganisms may pose a threat to the health of consumers. Because of that, in this review, we emphasize the risk that may be associated with the lack of appropriate studies of the quality of the supplements.
Since honeybee pollen is considered a “perfectly complete food” and is characterized by many beneficial properties (anti-inflammatory, antioxidant, anti-bacterial, etc.), it has begun to be used for therapeutic purposes. Consequently, there is a high need to develop methods for controlling its composition. A thorough bee pollen analysis can be very informative regarding its safety for consumption, the variability of its composition, its biogeographical origin, or harvest date. Therefore, in this study, two reliable and non-destructive spectroscopy methods, i.e., ED-XRF and ATR–FTIR, are proposed as a fast approach to characterize bee pollen. The collected samples were derived from apiaries located in west-central Poland. Additionally, some commercially available samples were analyzed. The applied methodology was optimized and combined with sophisticated chemometric tools. Data derived from IR analyses were also subjected to two-dimensional correlation spectroscopy. The developed ED-XRF method allowed the reliable quantification of eight macro- and micro-nutrients, while organic components were characterized by IR spectroscopy. Principal component analysis, cluster analysis, and obtained synchronous and asynchronous maps allowed the study of component changes occurring dependently on the date and location of harvest. The proposed approach proved to be an excellent tool to monitor the variability of the inorganic and organic content of bee pollen.
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