Alternaria toxins as casein kinase 2 inhibitors and possible consequences for estrogenicity: a hybrid in silico/in vitro study

Emerging mycotoxins produced by Alternaria spp. were previously reported to exert cytotoxic, genotoxic, but also estrogenic effects in human cells. The involved mechanisms are very complex and not fully elucidated yet. Thus, we followed an in silico target fishing approach to extend knowledge on the possible biological targets underlying the activity of alternariol, taken as the signature compound of Alternaria toxins. Combining ligand-based screening and structure-based modeling, the ubiquitous casein kinase 2 (CK2) was identified as a potential target for the compound. This result was validated in a cell-free in vitro CK2 activity assay, where alternariol inhibited CK2 with an IC50 of 707 nM. As CK2 was recently discussed to influence estrogen receptor (ER) transcription and DNA-binding affinity, we assessed a potential impact on the mRNA levels of ERα or ERβ by qRT-PCR and on nuclear localization of the receptors by confocal microscopy, using estrogen-sensitive Ishikawa cells as a model. While AOH did not affect the transcription of ERα or ERβ, an increase in nuclear localization of ERα after incubation with 10 µM AOH was observed. However, this effect might be due to ER binding affinity and therefore estrogenicity of AOH. Furthermore, in silico docking simulation revealed not only AOH, but also a number of other Alternaria toxins as potential inhibitors of CK2, including alternariol monomethyl ether and the perylene quinone derivative altertoxin II (ATX-II). These findings were representatively confirmed in vitro for the perylene quinone derivative altertoxin II, which was found to inhibit the kinase with an IC50 of 5.1 µM. Taken together, we propose CK2 inhibition as an additional mechanism to consider in future studies for alternariol and several other Alternaria toxins

Cake Perception, Texture and Aroma Profile as Affected by Wheat Flour and Cocoa Replacement with Carob Flour

Carob flour has been used in the production of a wide range of functional food formulations such as bakery goods either as a natural sweetener or food ingredient that, when roasted, exerts a chocolate/cocoa-reminiscent flavor and color. The aim of the present study was twofold; firstly to study the effect of an increasing incorporation of roasted carob flour (0–70% flour basis) on the quality and sensory attributes of a conventional cocoa cake recipe and secondly to investigate the obtained volatile fraction responsible for the aroma by means of headspace solid phase microextraction (HS-SPME) technique coupled to gas chromatography/mass spectrometry (GC/MS) while comparing it with the control, cocoa-containing cake recipe. Thirty and fifty percent carob flour incorporation rendered cakes with acceptable texture and sensory attributes, comparable to the control cake recipe containing 20% cocoa. Similarity to cocoa aroma was attributed to a great number of odor active compounds mainly belonging to aldehydes, lactones, furan/pyran derivatives, and pyrrole derivatives