Application of supervised chemometric techniques and synchronized excitation-emission spectrofluorometric analysis for the verification of Maltese extra virgin olive oils.

The authentication of virgin olive oil samples usually requires the use of sophisticated and very expensive analytical techniques. In this study, the potential of fluorescence spectroscopy for the authentication and discrimination of Maltese extra virgin olive oils was carried out using synchronized excitation-emission spectroscopy. Samples were collected from various producers around the Maltese islands. Synchronous excitation emission spectra were collected in the region of 240-750 nm with wavelength intervals of 10, 30, 60, 80 120 and 185 nm and subjected to several supervised chemometric procedures. Partial least square regression, linear discriminate analysis, and artificial neural network were used to define the origin of the Maltese olive oil against olive oils derived from other neighboring countries in the Mediterranean region. After subjecting the spectroscopic data to different pre-treatments and variable selection procedures results obtained evidenced a higher classification accuracy. This accuracy and predictability were highly dependent on the wave interval used and on the chemometric method used, however it was found that in general spectra obtained using δ 10 nm were deemed the most appropriate, with PLS, ANN and LDA reaching 100% accuracy and predictability in discriminating Maltese extra virgin olive oils when using derivatized spectral transformations. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05371-x.

Determination of the Geographical Origin of Maltese Honey Using 1H NMR Fingerprinting.

The price of honey, as a highly consumed natural product, depends on its botanical source and its production environment, causing honey to be vulnerable to adulteration through mislabeling and inappropriate, fraudulent production. In this study, a fast and simple approach is proposed to tackle this issue through non-target one dimensional zg30 and noesypr1d 1H NMR fingerprint analysis, in combination with multivariate data analysis. Results suggest that composition differences in sugars, amino acids, and carboxylic acid were sufficient to discriminate between the tested honey of Maltese origin and that of non-local origin. Indeed, all chemometric models based on noesypr1d analysis of the whole fraction honey showed better prediction in geographical discrimination. The possibility of discrimination was further investigated through analysis of the honey's phenolic extract composition. The partial least squares models were deemed unsuccessful to discriminate, however, some of the linear discriminant analysis models achieved a prediction accuracy of 100%. Lastly, the best performing models of both the whole fraction and the phenolic extracts were tested on five samples of unknown geographic for market surveillance, which attained a high agreement within the models. Thus, suggesting the use of non-target 1H NMR coupled with the multivariate-data analysis and machine learning as a potential alternative to the current time-consuming analytical methods.

Toxic oligomers of the amyloidogenic HypF-N protein form pores in mitochondrial membranes.

Studies on the amyloidogenic N-terminal domain of the E. coli HypF protein (HypF-N) have contributed significantly to a detailed understanding of the pathogenic mechanisms in neurodegenerative diseases characterised by the formation of misfolded oligomers, by proteins such as amyloid-ß, α-synuclein and tau. Given that both cell membranes and mitochondria are increasingly recognised as key targets of oligomer toxicity, we investigated the damaging effects of aggregates of HypF-N on mitochondrial membranes. Essentially, we found that HypF-N oligomers characterised by high surface hydrophobicity (type A) were able to trigger a robust permeabilisation of mito-mimetic liposomes possessing cardiolipin-rich membranes and dysfunction of isolated mitochondria, as demonstrated by a combination of mitochondrial shrinking, lowering of mitochondrial membrane potential and cytochrome c release. Furthermore, using single-channel electrophysiology recordings we obtained evidence that the type A aggregates induced currents reflecting formation of ion-conducting pores in mito-mimetic planar phospholipid bilayers, with multi-level conductances ranging in the hundreds of pS at negative membrane voltages. Conversely, HypF-N oligomers with low surface hydrophobicity (type B) could not permeabilise or porate mitochondrial membranes. These results suggest an inherent toxicity of membrane-active aggregates of amyloid-forming proteins to mitochondria, and that targeting of oligomer-mitochondrial membrane interactions might therefore afford protection against such damage.

Application of ATR-FT-MIR for Tracing the Geographical Origin of Honey Produced in the Maltese Islands.

Maltese honey has been produced, marketed, and sold as an exclusive local gourmet food product for countless years. Yet, thus far, no study has evaluated the individuality of this local food product. The evaluation of the parameters and properties which characterise the provenance and floral source of honey have been the subject of various studies worldwide, owing to the price and potential beneficial properties of this food product. Models analysing the potential of attenuated total reflection mid-infrared (ATR-FT-MIR) spectroscopy in discriminating and classifying local honey from that of foreign origin were investigated using 21 Maltese honey samples and 49 honey samples collected from abroad (Sicily, Greece, Sweden, Italy, France, Estonia and other samples of mixed geographical origin). Through a combination of spectroscopic techniques, spectral transformations, variable selection and partial least squares discriminant analysis (PLS-DA), chemometric models which successfully classified the provenance of local and non-local honey were developed. The results of these models were also corroborated with other classification and pattern recognition techniques, such as linear discriminate analysis (LDA), support vector machines (SVM) and feed-forward artificial neural networks (FF-ANN).

Application of 1H and 13C NMR Fingerprinting as a Tool for the Authentication of Maltese Extra Virgin Olive Oil.

The application of 1H and 13C nuclear magnetic resonance (NMR) in conjunction with chemometric methods was applied for the discrimination and authentication of Maltese extra virgin olive oils (EVOOs). A total of 65 extra virgin olive oil samples, consisting of 30 Maltese and 35 foreign samples, were collected and analysed over four harvest seasons between 2013 and 2016. A preliminary examination of 1H NMR spectra using unsupervised principle component analysis (PCA) models revealed no significant clustering reflecting the geographical origin. In comparison, PCA carried out on 13C NMR spectra revealed clustering approximating the geographical origin. The application of supervised methods, namely partial least squares discriminate analysis (PLS-DA) and artificial neural network (ANN), on 1H and 13C NMR spectra proved to be effective in discriminating Maltese and non-Maltese EVOO samples. The application of variable selection methods significantly increased the effectiveness of the different classification models. The application of 13C NMR was found to be more effective in the discrimination of Maltese EVOOs when compared to 1H NMR. Furthermore, results showed that different 1H NMR pulse methods can greatly affect the discrimination of EVOOs. In the case of 1H NMR, the Nuclear Overhauser Effect (NOESY) pulse sequence was more informative when compared to the zg30 pulse sequence, since the latter required extensive spectral manipulation for the models to reach a satisfactory level of discrimination.

The First Identification of the Uniqueness and Authentication of Maltese Extra Virgin Olive Oil Using 3D-Fluorescence Spectroscopy Coupled with Multi-Way Data Analysis.

The potential application of multivariate three-way data analysis techniques, namely parallel factor analysis (PARAFAC) and discriminant multi-way partial least squares regression (DN-PLSR), on three-dimensional excitation emission matrix (3D-EEM) fluorescent data were used to identify the uniqueness and authenticity of Maltese extra virgin olive oil (EVOO). A non-negativity constrained PARAFAC model revealed that a four-component model provided the most appropriate solution. Examination of the extracted components in mode 2 and 3 showed that these belonged to different fluorophores present in extra virgin olive oil. Application of linear discriminate analysis (LDA) and binary logistic regression analysis on the concentration of the four extracted fluorophores, showed that it is possible to discriminate Maltese EVOOs from non-Maltese EVOOs. The application of DN-PLSR provided superior means for discrimination of Maltese EVOOs. Further inspection of the extracted latent variables and their variable importance plots (VIPs) provided strong proof of the existence of four types of fluorophores present in EVOOs and their potential application for the discrimination of Maltese EVOOs.

The effect of turmeric (Curcumin) supplementation on cytokine and inflammatory marker responses following 2 hours of endurance cycling.

BACKGROUND: Endurance exercise induces IL-6 production from myocytes that is thought to impair intracellular defence mechanisms. Curcumin inhibits NF-κB and activator protein 1, responsible for cytokine transcription, in cell lines. The aim of this study was to investigate the effect of curcumin supplementation on the cytokine and stress responses following 2 h of cycling. METHODS: Eleven male recreational athletes (35.5 ± 5.7 years; Wmax 275 ± 6 W; 87.2 ± 10.3 kg) consuming a low carbohydrate diet of 2.3 ± 0.2 g/kg/day underwent three double blind trials with curcumin supplementation, placebo supplementation, and no supplementation (control) to observe the response of serum interleukins (IL-6, IL1-RA, IL-10), cortisol, c-reactive protein (CRP), and subjective assessment of training stress. Exercise was set at 95% lactate threshold (54 ± 7% Wmax) to ensure that all athletes completed the trial protocol. RESULTS: The trial protocol elicted a rise in IL-6 and IL1-RA, but not IL-10. The supplementation regimen failed to produce statistically significant results when compared to placebo and control. IL-6 serum concentrations one hour following exercise were (Median (IQR): 2.0 (1.8-3.6) Curcumin; 4.8 (2.1-7.3) Placebo; 3.5 (1.9-7.7) Control). Differences between supplementation and placebo failed to reach statistical significance (p = 0.18) with the median test. Repeated measures ANOVA time-trial interaction was at p = 0.06 between curcumin supplementation and placebo. A positive correlation (p = 0.02) between absolute exercise intensity and 1 h post-exercise for IL-6 concentration was observed. Participants reported "better than usual" scores in the subjective assessment of psychological stress when supplementing with curcumin, indicating that they felt less stressed during training days (p = 0.04) compared to placebo even though there was no difference in RPE during any of the training days or trials. CONCLUSION: The limitations of the current regimen and trial involved a number of factors including sample size, mode of exercise, intensity of exercise, and dose of curcumin. Nevertheless these results provide insight for future studies with larger samples, and multiple curcumin dosages to investigate if different curcumin regimens can lead to statistically different interleukin levels when compared to a control and placebo.

Identification of the plant origin of the botanical biomarkers of Mediterranean type propolis.

Propolis is a honeybee product which bees produce by collecting resins from various botanical sources. The chemical composition of propolis is directly dependant on the availability of resinous plant materials in different geographic regions. This study was undertaken to evaluate the resinous plant sources used by bees to produce Mediterranean type propolis. Although this propolis type has already been the subject of numerous studies, its major botanical source had not yet been identified. In this study, using GC-MS analysis, we identify the resin of the common cypress, Cupressus_sempervirens, as the major plant source of the characteristic diterpene fingerprint profile of Mediterranean propolis.