Food Risk Analysis: Towards a Better Understanding of "Hazard" and "Risk" in EU Food Legislation.

For risk communication, it is important to understand the difference between "hazard" and "risk". Definitions can be found in Codex Alimentarius and the European Union (EU) General Food Regulation (EC) No. 178/2002. The use of these terms as synonyms or their interchange is a recurrent issue in the area of food safety, despite awareness-raising messages sent by EFSA (European Food Safety Authority) and other interested entities. A quick screening of the EU's food regulations revealed several inconsistencies. Hence, it was considered necessary to further investigate if regulations could act as a source for this problem. A software tool was developed to support the detection and listing of inconsistent translations of "hazard" and "risk" in certain EU food regulations. Subsequently, native-speaking experts working in food safety from each EU country were asked to provide their individual scientific opinion on the prepared list. All data were statistically analysed after applying numerical scores (1-5) describing different levels of consistency. Results showed that the most common problem was the interchange of "hazard" with "risk" and vice versa. This lack of consistency can create confusion that can further translate into misjudgments at food risk assessment and communication levels.

Proteomic data and structure analysis combined reveal interplay of structural rigidity and flexibility on selectivity of cysteine cathepsins.

Addressing the elusive specificity of cysteine cathepsins, which in contrast to caspases and trypsin-like proteases lack strict specificity determining P1 pocket, calls for innovative approaches. Proteomic analysis of cell lysates with human cathepsins K, V, B, L, S, and F identified 30,000 cleavage sites, which we analyzed by software platform SAPS-ESI (Statistical Approach to Peptidyl Substrate-Enzyme Specific Interactions). SAPS-ESI is used to generate clusters and training sets for support vector machine learning. Cleavage site predictions on the SARS-CoV-2 S protein, confirmed experimentally, expose the most probable first cut under physiological conditions and suggested furin-like behavior of cathepsins. Crystal structure analysis of representative peptides in complex with cathepsin V reveals rigid and flexible sites consistent with analysis of proteomics data by SAPS-ESI that correspond to positions with heterogeneous and homogeneous distribution of residues. Thereby support for design of selective cleavable linkers of drug conjugates and drug discovery studies is provided.

Implementation of food matrix effects into chemical food contaminant risk assessment.

Food risk assessment plays an important role in protecting public health worldwide. Stakeholders involved in food risk assessment, such as national authorities, agencies, non-governmental organisations (NGOs), industry and consumers, need to properly understand the terminology of food risk assessment effectively. In this respect, the first part of the EU-FORA work programme (WP1) aimed to provide insights into the actual translation of two essential terms used in food risk assessment. 'Hazard' and 'risk' were first identified and compared between the English version of various food regulations and their equivalents in the national legislation of EU Member States. The comparison and critical evaluation revealed several inconsistencies. These inconsistencies could lead to misinterpretations, followed by errors in conducting risk assessments or communicating risks. We recommend that consistency is restored and maintained so that the message is properly communicated. The second part of the work programme (WP2) was focused on a specific area within chemical risk assessment (CRA). In this context, special attention was given to the impact of the food matrix on the bioaccessibility and bioavailability of heavy metals and metalloids. After collection and careful selection of data from scientific journals, a database with information on the bioaccessibility and bioavailability of cadmium (Cd), lead (Pb), mercury (Hg) and arsenic (As) in different food matrices was created for future statistical analyses related to dietary exposure.

Mechanisms Applied by Protein Inhibitors to Inhibit Cysteine Proteases.

Protein inhibitors of proteases are an important tool of nature to regulate and control proteolysis in living organisms under physiological and pathological conditions. In this review, we analyzed the mechanisms of inhibition of cysteine proteases on the basis of structural information and compiled kinetic data. The gathered structural data indicate that the protein fold is not a major obstacle for the evolution of a protease inhibitor. It appears that nature can convert almost any starting fold into an inhibitor of a protease. In addition, there appears to be no general rule governing the inhibitory mechanism. The structural data make it clear that the "lock and key" mechanism is a historical concept with limited validity. However, the analysis suggests that the shape of the active site cleft of proteases imposes some restraints. When the S1 binding site is shaped as a pocket buried in the structure of protease, inhibitors can apply substrate-like binding mechanisms. In contrast, when the S1 binding site is in part exposed to solvent, the substrate-like inhibition cannot be employed. It appears that all proteases, with the exception of papain-like proteases, belong to the first group of proteases. Finally, we show a number of examples and provide hints on how to engineer protein inhibitors.

Changes of poultry faecal microbiota associated with Clostridium difficile colonisation.

Bacterial, fungal and archaeal microbiota was analysed in 143 chicken faecal samples from a single poultry farm. After DHPLC (denaturing high performance liquid chromatography) 15 bacterial groups, 10 fungal groups and a single archaeal species were differentiated. Samples were grouped into two clusters with significantly different frequencies of C. difficile positive and negative samples in each cluster. Acidaminococcus intestini, described here for the first time as a part of poultry faecal microbiota, was significantly more likely present in C. difficile negative samples, while presence/absence of some other microorganisms (Enterococcus cecorum, Lactobacillus galinarum, Moniliella sp. and Trichosporon asahii) was close to significance. Two other groups not reported previously for poultry, Coprobacillus sp. and Turicibacter sp. did not differ significantly between C. difficile positive and negative samples. Differences in microbiota diversity depend on animal age, but not on the presence of C. difficile. With machine learning (WEKA J48) we have defined specific combinations of microbial groups predictive for C. difficile colonisation. Microbial groups associated with C. difficile colonisation in poultry are different than those reported for humans and include bacteria as well as fungi. Also with this approach A. intestini was found to be most strongly related to C. difficile negative samples.