The high temperatures used in the production of milk may induce modifications in proteins structure. Due to occurrence of the Maillard reaction, lactose binds lysine residues in proteins, affecting the nutritional value. Milk is also an important source of allergenic proteins (i.e., caseins, β-lactoglobulin and α-lactalbumin). Thus, this modification may also affect the allergenicity of these proteins. Focusing on milk whey proteins, a screening on different Ultra High Temperatures (UHT) and pasteurized milk samples was performed to identify lactosylation sites, in particular in protein known epitopes, and to verify the correlation between lactosylation and the harshness of the treatment. Whey proteins were extracted from milk samples after caseins precipitations at pH 4.6 and, after chymotryptic and tryptic in solution digestion, peptides were analysed by UPLC-MS and LTQ-Orbitrap. Results show the presence of lactosylated lysine residues in several known epitopes. Then, a β-lactoglobulin epitope was selected and synthesized by solid phase synthesis followed by in solution lactosylation, obtaining high reaction yields and purities. The synthesis of lactosylated allergenic epitopes, described here for the first time, is a useful tool for further studies on the technological impacts on food allergenicity.

Hearing loss (HL) affects millions of people worldwide. While epidemiological data from Western countries are already available, this information is still lacking for many developing countries and rural communities. Here we report, for the first time, a study on the frequency of HL in a series of rural communities located along the Silk Road. Study design: Four hundred and ninety-six subjects (236 males and 260 females ranging from eight to 84 years of age) selected in non-random, convenience samples from rural communities belonging to Terra Madre Organization have been enrolled: 228 from Georgia and Azerbaijan (Caucasus region), 151 from Uzbekistan and Kazakhstan (Central Asia), and 117 from Pair (Tajikistan). Subjects underwent pure-tone audiometry at 0.25, 0.5, 1, 2, 4, 6, and 8 kHz; pure-tone average (PTA) values were determined. Results: The overall HL frequency, ranging from 9% to 18%, in the investigated communities located along the Silk Road is higher than that reported for WHO European region countries but comparable with that reported for WHO South-East Asian region countries and other developing countries. Interestingly, with regard to the impact of age, two different disease behaviours have been identified in all the tested communities – one for males and the other for females. In females HL starts at all PTAs between 30 and 40 years of age. In contrast, in males age starts to affect hearing at high frequencies in young adulthood (20–25 years old), but later (50–60 years old) at low and medium frequencies. Conclusion: Despite the difficulty in reaching rural communities, mainly located in remote places, and the need to perform further studies using a larger sample size, recent data provide new information and will contribute to a better definition of HL worldwide frequency

The main coffee diterpenes cafestol, kahweol, and 16-O-methylcafestol, present in the bean lipid fraction, are mostly esterified with fatty acids. They are believed to induce dyslipidaemia and hypercholesterolemia when taken with certain types of coffee brews. The study of their binding to serum albumins could help explain their interactions with biologically active xenobiotics. We investigated the interactions occurring between cafestol and 16-O-methylcafestol palmitates with Bovine Serum Albumin (BSA), Human Serum Albumin (HSA), and Fatty Free Human Serum Albumin (ffHSA) by means of circular dichroism and fluorimetry. Circular Dichroism (CD) revealed a slight change (up to 3%) in the secondary structure of fatty-free human albumin in the presence of the diterpene esters, suggesting that the aliphatic chain of the palmitate partly occupies one of the fatty acid sites of the protein. A warfarin displacement experiment was performed to identify the binding site, which is probably close but not coincident with Sudlow site I, as the affinity for warfarin is enhanced. Fluorescence quenching titrations revealed a complex behaviour, with Stern–Volmer constants in the order of 10³–10⁴ Lmol⁻¹. A model of the HSA-warfarin-cafestol palmitate complex was obtained by docking, and the most favourable solution was found with the terpene palmitate chain inside the FA4 fatty acid site and the cafestol moiety fronting warfarin at the interface with site I.