In this paper the effect of aflatoxin B1, ochratoxin A and zearalenon on morphology, growth parameters and metabolic activity of yeasts Saccharomyces cerevisiae, Saccharomyces uvarum, Candida utilis and Kluyveromyces marxianus was determined. The results showed that the three mycotoxins affected the morphology of all these yeasts, primarily the cell diameter, but not their final cell count. Fourier transform infrared spectroscopy showed that the yeast membranes bound the mycotoxins, C. utilis in particular. The cell membranes of most yeasts underwent denaturation, except S. uvarum exposed to ochratoxin A and zearalenone. In the early stage of fermentation, all mycotoxin-exposed yeasts had lower metabolic activity and biomass growth than controls, but fermentation products and biomass concentrations reached the control levels by the end of the fermentation, except for C. utilis exposed to 20 µg/mL of zearalenone. The adaptive response to mycotoxins suggests that certain yeasts could be used to control mycotoxin concentrations in the production of fermented food and beverages.

Maceration parameters such as alcohol and mistletoe content were studied to obtain high-quality macerates to produce biska, a traditional Istrian herbal spirit. An ethanol–water solution containing 25, 40, 55, and 70 v/v% ethanol and 20, 40, and 80 g/L mistletoe was macerated and pH, total acids, extract content, and color parameters were measured. Volatile compounds were determined by GC/MS. More than 150 different volatile compounds were detected. The composition of the ethanol–water solution and mistletoe content affects the values of total extract, total acids, and color. Samples with lower alcohol content have a higher number of different acids, aldehydes, alcohols, esters, and ketones, while macerates with higher alcohol content are rich in hydrocarbons and terpenes. Ethyl decanoate; ethyl hexynoate; ethyl octanoate; benzaldehyde; hexanal; octanal; and the terpene compounds limonene, cymene, menthone, humulene, eucalyptol, linalol, and borneol contribute significantly to the aroma of the macerates due to a low odor threshold. Ylangene and α- and β-bourbonene are tentatively determined new terpene molecules that are not found in the mistletoe literature, as well as thujone, which was also detected. For the preparation of aromatic macerates, 40–50 v/v% ethanol and about 40 g/L mistletoe should be used.

Biska, a traditional Istrian herbal spirit, is produced by macerating mistletoe leaves and twigs in a water–ethanol base, usually grape marc spirit. The aim of this study was to determine the optimal mistletoe maceration parameters (ethanol content, concentration of plant material, and duration) for the production of biska, which have not yet been the subject of scientific research. The prepared mistletoe macerates and the collected biska samples were subjected to phenolic compound characterization, including total phenolic content (TPC), total flavonoid content (TFC), and polyphenolic profile, by ultra-high-performance liquid chromatography coupled with mass spectrometry (UPLC-ESI-MS²). In addition, the macerates and the biska samples were analyzed for their biological properties: antioxidant and antiproliferative activity. The results showed that both the macerates and the biska samples contain phenolic compounds of mistletoe such as phenolic acids, flavonoids, and coumarins, which contribute to the observed biological activities—significant antioxidant activity and antiproliferative effect. By applying the maceration parameters, which yielded the highest content of phenolic compounds as well as volatile aromatic compounds (40–80 g/L of mistletoe, 40–55% v/v ethanol, and 21 days’ maceration time), the production process can be improved to produce biska with better aromatic and biological properties.

Research background. During the current SARS-CoV2 pandemic, as well as earlier SARS and MERS epidemics, it has been observed that COVID19-positive women on average tend to have milder symptoms and lower fatality rates than men. There is a number of differences between the sexes known to contribute to different immune responses and severity of the disease, one being the effect of estrogen via estrogen receptor signalling. We wondered if estrogen might also affect the SARS-CoV2 more directly, perhaps by binding to the surface glycoprotein (S protein), thus possibly reducing its infectivity. Experimental approach. To assess whether there is a possibility for estrogen binding on the SARS- CoV2 S protein, we used BLAST and HHpred to compare protein sequences of S protein and human estrogen receptor beta, while 3D structures of a potential estrogen binding site and an active site of estrogen receptor beta were visualized and compared using PyMOL. Results and conclusions. By comparing the sequence of SARS-CoV2 S protein with the human estrogen receptor beta, we identified a potential estrogen binding site on S protein and further determined that it also shares notable similarities with the active site of ER beta when observed in 3D structure of their respective proteins. As a control, SARS-CoV2 S protein was compared with the human androgen receptor, and no such similarities were found. The potential estrogen binding site is part of coronavirus S2 superfamily domain, which is involved in host-virus membrane fusion during infection and appears to be conserved throughout the Coronaviridae family. Novelty and scientific contribution: This preliminary communication shows that SARS- CoV2 S protein features a potential estrogen binding site. Hopefully, this will prompt a more comprehensive study on the possibilities of estrogen binding on the S protein and the effect this might confer on the virus infectivity.