Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease with no therapeutic consensus. Oxidation and inflammation are hallmarks in the progression of this complex disease, which also involves interactions between the genetic background and the environment. Mastiha is a natural nutritional supplement known to possess antioxidant and anti-inflammatory properties. This study investigated how a 6-month Mastiha supplementation (2.1 g/day) could impact the antioxidant and inflammatory status of patients with NAFLD, and whether genetic variants significantly mediate these effects. We recruited 98 patients with obesity (BMI ≥ 30 kg/m²) and NAFLD and randomly allocated them to either the Mastiha or the placebo group for 6 months. The anti-oxidative and inflammatory status was assessed at baseline and post-treatment. Genome-wide genetic data was also obtained from all participants, to investigate gene-by-Mastiha interactions. NAFLD patients with severe obesity (BMI > 35kg/m²) taking the Mastiha had significantly higher total antioxidant status (TAS) compared to the corresponding placebo group (P value=0.008). We did not observe any other significant change in the investigated biomarkers as a result of Mastiha supplementation alone. We identified several novel gene-by-Mastiha interaction associations with levels of cytokines and antioxidant biomarkers. Some of the identified genetic loci are implicated in the pathological pathways of NAFLD, including the lanosterol synthase gene (LSS) associated with glutathione peroxidase activity (Gpx) levels, the mitochondrial pyruvate carrier-1 gene (MPC1) and the sphingolipid transporter-1 gene (SPNS1) associated with hemoglobin levels, the transforming growth factor‐beta‐induced gene (TGFBI) and the micro-RNA 129-1 (MIR129-1) associated with IL-6 and the granzyme B gene (GZMB) associated with IL-10 levels. Within the MAST4HEALTH randomized clinical trial (NCT03135873, www.clinicaltrials.gov) Mastiha supplementation improved the TAS levels among NAFLD patients with severe obesity. We identified several novel genome-wide significant nutrigenetic interactions, influencing the antioxidant and inflammatory status in NAFLD.Clinical Trial Registration: ClinicalTrials.gov, identifier NCT03135873.

Life history traits determine the persistence and reproduction of each species. Factors that can affect life history traits are numerous and can be of different origin. We investigated the influence of population origin and heavy metal exposure on microbiota diversity and two life history traits, egg-to-adult viability and developmental time, in Drosophila melanogaster and Drosophila subobscura, grown in the laboratory on a lead (II) acetate-saturated substrate. We used 24 samples, 8 larval and 16 adult samples (two species × two substrates × two populations × two sexes). The composition of microbiota was determined by sequencing (NGS) of the V3–V4 variable regions of the 16S rRNA gene. The population origin showed a significant influence on life history traits, though each trait in the two species was affected differentially. Reduced viability in D. melanogaster could be a cost of fast development, decrease in Lactobacillus abundance and the presence of Wolbachia. The heavy metal exposure in D. subobscura caused shifts in developmental time but maintained the egg-to-adult viability at a similar level. Microbiota diversity indicated that the Komagataeibacter could be a valuable member of D. subobscura microbiota in overcoming the environmental stress. Research on the impact of microbiota on the adaptive response to heavy metals and consequently the potential tradeoffs among different life history traits is of great importance in evolutionary research.

Coprinellus disseminatus (Pers.) J.E. Lange 1938 is an edible saprotrophic fungal species with bioactive potential which is not extensively investigated, mostly due to its small size. Accordingly, the aim of this study was to determine chemical and biological properties of crude ethanol (CdEtOH) and water extracts (CdAq). The bioactivity screens included antioxidant and antiproliferative (human breast MCF-7 cancer cell-line; MTT and SRB assays) activity. In addition, CdAq and CdEtOH were primarily characterized by UV-VIS spectrophotometry, with an aim of determining its chemical composition (total phenol and flavonoid contents). The best antioxidant activities were obtained for hydroxyl radicals (OH∙) and superoxide anion radical (SOA) assays for both crude extracts (IC50 =4.02 μg/ml for CdAq and IC50 =1.40 μg/ml for CdEtOH) while total phenolic (TP) and total flavonoid (TF) content was higher in CdEtoH than in CdAq. Both crude extracts have shown activity against MCF-7 cell line although CdEtOH showed better effect than CdAq extract after 72h in MTT and SRB assay, reaching IC50 at 217.90 and 205.90 μg/ml, respectively. Based on obtained results, the saprotrophic fungal species Coprinellus disseminatus (Pers.) J.E. Lange 1938 could be considered as a good potential alternative source of nutraceuticals and biologically active compounds.

The composition of microbiota affects different traits of Drosophila throughout its life cycle and represents an important part of the adaptive response to environmental changes, such as increased levels of accumulated heavy metals in their habitats. We investigated the effects of lead added to the feeding substrate on the microbiota diversity in the fruit flies, Drosophila melanogaster and Drosophila subobscura. We compared the bacterial diversity of wild-caught flies and their progeny reared under standard laboratory conditions. We analyzed the shifts in bacterial diversity in the experimental groups of flies reared for 13 generations on standard and lead(II) acetate (Pb(CH3COO)2), Pb acetate-saturated substrates. Identification of the main bacterial genera was performed by next-generation sequencing (NGS) of the V3-V4 variable regions of the 16S rRNA gene. Results indicate significant species-specific differences in bacterial composition between natural and laboratory populations, and between the substrates. Diversity increased in both species under prolonged exposure to lead-polluted substrate, suggesting a potential adaptive response to an environmental stress.

Whereas the etiology of non-alcoholic fatty liver disease (NAFLD) is complex, the role of nutrition as a causing and preventive factor is not fully explored. The aim of this study is to associate dietary patterns with magnetic resonance imaging (MRI) parameters in a European population (Greece, Italy, and Serbia) affected by NAFLD. For the first time, iron-corrected T1 (cT1), proton density fat fraction (PDFF), and the liver inflammation fibrosis score (LIF) were examined in relation to diet. A total of 97 obese patients with NAFLD from the MAST4HEALTH study were included in the analysis. A validated semi-quantitative food frequency questionnaire (FFQ) was used to assess the quality of diet and food combinations. Other variables investigated include anthropometric measurements, total type 2 diabetes risk, physical activity level (PAL), and smoking status. Principal component analysis (PCA) was performed to identify dietary patterns. Six dietary patterns were identified, namely “High-Sugar”, “Prudent”, “Western”, “High-Fat and Salt”, “Plant-Based”, and “Low-Fat Dairy and Poultry”. The “Western” pattern was positively associated with cT1 in the unadjusted model (beta: 0.020, p-value: 0.025) and even after adjusting for age, sex, body mass index (BMI), PAL, smoking, the center of the study, and the other five dietary patterns (beta: 0.024, p-value: 0.020). On the contrary, compared with low-intake patients, those with medium intake of the “Low-Fat Dairy and Poultry” pattern were associated with lower values of cT1, PDFF, and LIF. However, patients with a “Low-Fat Dairy and Poultry” dietary pattern were negatively associated with MRI parameters (cT1: beta: −0.052, p-value: 0.046, PDFF: beta: −0.448, p-value: 0.030, LIF: beta: −0.408, p-value: 0.025). Our findings indicate several associations between MRI parameters and dietary patterns in NAFLD patients, highlighting the importance of diet in NAFLD.

In response to the growing demand for clean-label preservatives, this study investigates the potential of Laetiporus sulphureus, an edible polypore mushroom, as a multifunctional additive in cooked sausages. The ethanolic extract of L. sulphureus (LsEtOH) was evaluated for its chemical composition, antioxidant capacity, and antimicrobial activity. Leucine (12.4 ± 0.31 mg/g d.w.) and linoleic acid (68.6%) were identified as the dominant essential amino acid and fatty acid. LsEtOH exhibited strong antioxidant activity, with IC₅₀ values of 215 ± 0.05 µg/mL (DPPH•), 182 ± 0.40 µg/mL (NO•), and 11.4 ± 0.01 µg/mL (OH•), and showed a selective inhibition of Gram-positive bacteria, particularly Staphylococcus aureus (MIC/MBC: 0.31/0.62 mg/mL). In cooked sausages treated with 0.05 mg/kg of LsEtOH, lipid peroxidation was reduced (TBARS: 0.26 mg MDA/kg compared to 0.36 mg MDA/kg in the control), microbial growth was suppressed (33.3 ± 15.2 CFU/g in the treated sample compared to 43.3 ± 5.7 CFU/g in the control group), and color and pH were stabilized over 30 days. A sensory evaluation revealed minor flavor deviations due to the extract’s inherent aroma. Encapsulation and consumer education are recommended to enhance acceptance. This is the first study to demonstrate the efficacy of L. sulphureus extract as a natural preservative in a meat matrix, supporting its application as a clean-label additive for shelf life and safety improvement.