Parkin Mutation Affects Clock Gene-Dependent Energy Metabolism.

Growing evidence highlights a tight connection between circadian rhythms, molecular clockworks, and mitochondrial function. In particular, mitochondrial quality control and bioenergetics have been proven to undergo circadian oscillations driven by core clock genes. Parkinson's disease (PD) is a chronic neurodegenerative disease characterized by a selective loss of dopaminergic neurons. Almost half of the autosomal recessive forms of juvenile parkinsonism have been associated with mutations in the PARK2 gene coding for parkin, shown to be involved in mitophagy-mediated mitochondrial quality control. The aim of this study was to investigate, in fibroblasts from genetic PD patients carrying parkin mutations, the interplay between mitochondrial bioenergetics and the cell autonomous circadian clock. Using two different in vitro synchronization protocols, we demonstrated that normal fibroblasts displayed rhythmic oscillations of both mitochondrial respiration and glycolytic activity. Conversely, in fibroblasts obtained from PD patients, a severe damping of the bioenergetic oscillatory patterns was observed. Analysis of the core clock genes showed deregulation of their expression patterns in PD fibroblasts, which was confirmed in induced pluripotent stem cells (iPSCs) and induced neural stem cells (iNSCs) derived thereof. The results from this study support a reciprocal interplay between the clockwork machinery and mitochondrial energy metabolism, point to a parkin-dependent mechanism of regulation, and unveil a hitherto unappreciated level of complexity in the pathophysiology of PD and eventually other neurodegenerative diseases.

Bioactive compounds from orange epicarp to enrich fish burgers.

BACKGROUND: The orange industry produces considerable amounts of by-products, traditionally used for animal feed or fuel production. Most of these by-products could be used as functional ingredients. To assess the potential food application of orange epicarp, different percentages of micro-encapsulated orange extract were added to fresh fish burgers. Then, an in vitro digestion was also carried out, before and after micro-encapsulation, to measure the bio-accessibility of the active compounds. RESULTS: A significant increase of bio-accessibility of bioactive compounds has been observed in the orange epicarp extract after micro-encapsulation by spray-drying. From the sensory point of view, the fish sample enriched with 50 g kg-1 micro-encapsulated extract was the most comparable to the control burger, even if it showed a higher phenolic, flavonoid and carotenoid bio-accessibility. CONCLUSION: Orange epicarp may be used as a food additive to enhance the health content of food products. The micro-encapsulation is a valid technique to protect the bioactive compounds and increase their bio-accessibility. © 2017 Society of Chemical Industry.

Durum wheat breads 'high in fibre' and with reduced in vitro glycaemic response obtained by partial semolina replacement with minor cereals and pulses.

The aim of this study was to develop fortified breads of durum wheat semolina (DW) partially substituted at 10%, 15%, 20% and 30% with white sorghum or yellow pea wholemeal flours (WS, YP, respectively) or using wholemeal flour from a natural mixture of rye and durum wheat (RDW). The physico-chemical composition of the raw materials, rheological properties of dough, the bread quality characteristics, glycaemic index and sensory quality of bread were examined. Compared to 100% DW, 100% YP had twice the protein levels, RDW had almost three times the dietary fibre, while the colorimetric indices for WS and YP flour addition showed increased redness (for RDW) and similar yellowness (for YP). With respect to the control dough (100% DW), RDW and addition of WS or YP produced showed negative impact on water absorption (RDW, WS), stability (RDW, YP), dough strength (RDW, WS, YP), tenacity and extensibility ratio (RDW, YP), loaf volume (RDW, WS, YP), yellowness (RDW, WS) and sensory acceptance of bread (RDW, WS). However, these changes were counterbalanced by higher dietary fibre and lower glycaemic index of the breads, especially for RDW and at high additions of WS and YP (20-30%). Moreover, breads fortified with YP were better in terms of colour and overall acceptability scores.

Optimization and characterization of gluten-free spaghetti enriched with chickpea flour.

This work was focused on the optimization and characterization of maize-based spaghetti fortified with chickpea flour. To the aim, the study has been organized in two subsequent trials. In the first one, the chickpea flour amount added to the spaghetti was continuously increased until the overall sensory quality of pasta reached its sensory threshold. Spaghetti samples loaded with 15% chickpea flour showed poor elasticity and increased firmness, so this concentration represented the highest chickpea flour concentration to be used. The second experimental step was aimed to improve the overall sensory quality of the enriched spaghetti by means of hydrocolloids as pectin, guar flour and agar. Final pasta was characterized for the nutritional composition, the glycemic response and the main quality attributes. The best results were obtained by the addition of guar flour.

Effect of durum wheat cultivars on physico-chemical and sensory properties of spaghetti.

BACKGROUND: Mixtures of different cultivars provide semolina with proper processing properties but not always good nutritional properties. In this study, the effects of mono-varietal cultivars of durum wheat on pasta quality were evaluated in order to find a good balance between nutritional and sensory properties of the final product. RESULTS: Durum wheat spaghetti was manufactured using semolina from six mono-varietal cultivars. A commercially available semolina mixture was also used to produce a control pasta sample. Instrumental (i.e. rheological and texture analysis), sensory (i.e. elasticity, firmness, adhesiveness) and nutritional (i.e. protein, ash and fibre content, glycaemic index) analyses were carried out. Results highlighted differences between selected cultivars. In particular, spaghetti obtained with Anco Marzio and Cappelli semolina (modern and old cultivars, respectively) showed the highest protein content and the lowest cooking loss, compared with the other samples. CONCLUSION: Spaghetti made with Cappelli semolina showed the lowest adhesiveness and the highest hardness; it recorded the best overall quality and presented the lowest glycaemic response. Hence, durum wheat pasta with a good balance between nutritional and cooking quality could be obtained from semolina based on mono-varietal cultivars. © 2013 Society of Chemical Industry.

Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells.

Background: Alzheimer's disease (AD) is a devastating neurodegenerative disease without guidelines for early diagnosis or personalized treatment. Previous studies have highlighted a crucial role of increasing phosphorylation levels of the amyloid precursor protein (APP) Tyr682 residue in predicting neuronal deficits in AD patients. However, the lack of a method for the identification and quantification of Tyr682 phosphorylation levels prevents its potential clinical applications. Methods: Here we report a method to identify and quantify APP Tyr682 phosphorylation levels in blood mononuclear cells of AD patients by tandem mass spectrometry (tMS). Results: This method showed excellent sensitivity with detection and quantification limits set respectively at 0.035 and 0.082 ng injected for the phosphorylated peptide and at 0.02 and 0.215 ng injected for the non-phosphorylated peptide. The average levels of both peptides were quantified in transfected HELA cells (2.48 and 3.53 ng/µg of protein, respectively). Preliminary data on 3 AD patients showed quantifiable levels of phosphorylated peptide (0.10-0.15 ng/µg of protein) and below the LOQ level of non-phosphorylated peptide (0.13 ng/µg of protein). Conclusion: This method could allow the identification of patients with increased APP Tyr682 phosphorylation and allow early characterization of molecular changes prior to the appearance of clinical signs.

Study on the Influence of Bio-Based Packaging System on Sodium Benzoate Release Kinetics.

The influence of film structure on the release kinetics of sodium benzoate (SB) from polymeric films is addressed in this study. In particular, four film structures were investigated, two monolayer and two multilayer systems. In particular, in one case, the active substance was uniformly distributed into a chitosan-based matrix, and in the other one, it was previously incorporated into alginate beads before dispersion in the chitosan film, thus realizing two types of monolayer films; on the other hand, the same chitosan film with SB encapsulated in alginate beads was used as the inner layer of a multilayer system constituted by two side films of alginate. The two alginate-based layers were made with two different thicknesses, thus producing a total of two multilayer systems. The release of SB from the above-mentioned films in water was studied by means of a UV/VIS spectrophotometer at 227 nm. A first-order kinetics-type equation was used to quantitatively describe the release data. Results suggest that the film structure strongly affected the release kinetics. In fact, monolayer films showed single-stage release kinetics, whereas the two investigated multilayer systems showed two-stage release kinetics. Further, the presence of alginate beads strongly affected the SB release, thus suggesting the potential of encapsulation to control the release mechanism of active compounds.

Effect of Chicken Bone Extracts on Metabolic and Mitochondrial Functions of K562 Cell Line.

BACKGROUND: Tetracyclines' use in intensive animal farming has raised some concerns regarding the biosafety for humans. Increasing evidences have revealed the presence of these drugs in processed animal by-products, such as bone, throughout the food chain. A potential off-target of tetracyclines is the bacterial-like mitochondrial translational machinery, thereby causing proteostatic alterations in mitochondrial DNA-encoded components of the oxidative phosphorylation system. METHODS: The Seahorse methodology, confocal microscopy imaging of mitochondrial potential and reactive oxygen species, and q-RT-PCR analysis of the expression of genes involved in mitochondrial biogenesis and mitophagy were carried out on human lymphoblast derived K562 cell line challenged with bone powder derived from chicken treated with or without oxytetracycline and pure oxytetracycline. RESULTS: A complex dose-dependent profile was attained with a low dosage of bone powder extracts causing a metabolic adaptation hallmarked by stimulation of the mitochondrial respiration and enhanced expression of mitochondriogenic factors in particular in cells challenged with oxytetracycline-free bone extract. Conversely, a higher dosage of bone powder extracts, regardless of their source, caused a progressive inhibition of mitochondrial respiration and glycolysis, ultimately leading to cell death. No significant effects of the pure oxytetracycline were observed. CONCLUSION: Bone powder, regardless of chicken treatment, contains and releases factors/chemicals responsible for the observed effects on energy metabolism. Quantitative differential effects appear to depend on biochemical alterations in the bone matrix caused by antibiotics rather than antibiotics themselves.

Use of purple durum wheat to produce naturally functional fresh and dry pasta.

In this study, the effects of different milling procedures (roller-milling vs. stone-milling) and pasta processing (fresh vs. dried spaghetti), and cooking on the antioxidant components and sensory properties of purple durum wheat were investigated. Milling and pasta processing were performed using one purple and one conventional non-pigmented durum wheat genotypes, and the end-products were compared with commercial pasta. The results show that the stone milling process preserved more compounds with high health value (total fibre and carotenoids, and in the purple genotype, also anthocyanins) compared to roller-milling. The drying process significantly (p<0.05) reduced the content of anthocyanins (21.42 µg/g vs. 46.32 µg/g) and carotenoids (3.77 µg/g vs. 4.04 µg/g) with respect to the pasteurisation process involved in fresh pasta production. The sensory properties of pasta from the purple genotype did not significantly differ from commercial wholemeal pasta, and its in vitro glycemic index was even lower. Thus, it is possible to consider this genetic material as a good ingredient for the production of functional foods from cereals naturally rich in bioactive compounds.

trans-Resveratrol, quercetin, (+)-catechin, and (-)-epicatechin content in south Italian monovarietal wines: relationship with maceration time and marc pressing during winemaking.

The concentrations of trans-resveratrol, (+)-catechin, (-)-epicatechin, and quercetin were evaluated by means of high-performance liquid chromatography-diode array detection in red wines obtained from Aglianico, Piedirosso, and Nerello Mascalese grapes. The trans-resveratrol and epicatechin concentrations did not differ significantly between experimental wines. The concentration of quercetin in Nerello Mascalese wines was more than twice that observed in Aglianico and Piedirosso wines. Nerello Mascalese wines also significantly differed from other wines in the (+)-catechin content, which was significantly higher than those found in the other two wines. During maceration, the maximum extraction of trans-resveratrol was reached after 12 days for Aglianico and Piedirosso, after which a decline was observed. On the contrary, in the case of Nerello Mascalese, the concentration of trans-resveratrol increased steadily throughout the whole maceration process. After 2 days of maceration, the maximum concentration of quercetin was observed in Aglianico must, whereas the maximum quercetin extraction was reached after 12 days for Piedirosso and 17 days for Nerello Mascalese. The maximum levels of (+)-catechin and (-)-epicatechin were generally observed after 12 days of maceration for all wines, although a decline of (-)-epicatechin occurred after maximum extraction in Aglianico and Piedirosso wines. Following marc pressing, a significant increase in the concentration of trans-resveratrol for Aglianico, (+)-catechin and (-)-epicatechin for Piedirosso, and (-)-epicatechin for Nerello Mascalese was observed.

Bone-marrow derived hematopoietic stem/progenitor cells express multiple isoforms of NADPH oxidase and produce constitutively reactive oxygen species.

Consolidated evidence highlights the importance of redox signalling in poising the balance between self-renewal and differentiation in adult stem cells. The present study shows that human hematopoietic stem/progenitor cells (HSCs) constitutively generate low levels of hydrogen peroxide whose production is inhibited by DPI, apocynin, catalase, and LY294002 and scarcely stimulated by PMA. Moreover, it is shown that HSCs express at the mRNA and protein levels the catalytic subunits of NOX1, NOX2, and NOX4 isoforms of the NADPH oxidase family along with the complete battery of the regulatory subunits p22, p40, p47, p67, rac1, rac2, NOXO1, and NOXA1 as well as the splicing variant NOX2s and that the three NOX isoforms are largely co-expressed in the same HSC. These findings are interpreted in terms of a positive feed-back mechanism of NOXs activation enabling a fine tuning of the ROS level to be possibly used in redox-mediated signalling for growth and differentiation of HSCs.

Hepatitis C virus protein expression causes calcium-mediated mitochondrial bioenergetic dysfunction and nitro-oxidative stress.

UNLABELLED: Hepatitis C virus (HCV) infection induces a state of oxidative stress that is more pronounced than that in many other inflammatory diseases. In this study we used well-characterized cell lines inducibly expressing the entire HCV open-reading frame to investigate the impact of viral protein expression on cell bioenergetics. It was shown that HCV protein expression has a profound effect on cell oxidative metabolism, with specific inhibition of complex I activity, depression of mitochondrial membrane potential and oxidative phosphorylation coupling efficiency, increased production of reactive oxygen and nitrogen species, as well as loss of the Pasteur effect. Importantly, all these effects were causally related to mitochondrial calcium overload, as inhibition of mitochondrial calcium uptake completely reversed the observed bioenergetic alterations. CONCLUSION: Expression of HCV proteins causes deregulation of mitochondrial calcium homeostasis. This event occurs upstream of further mitochondrial dysfunction, leading to alterations in the bioenergetic balance and nitro-oxidative stress. These observations provide new insights into the pathogenesis of hepatitis C and may offer new opportunities for therapeutic intervention.