The role of nutrition in multiple sclerosis: a story yet to be written.

The role of nutrition in multiple sclerosis (MS) is still unclear and MS therapy is not associated to a particular diet. In this review we show the molecular basis by which dietary factors and life style may exacerbate or ameliorate MS symptoms, by controlling both the metabolic and inflammatory pathways in the cell and the composition of gut microbiota. A persistent “Western style” diet may indeed lead to a dysbiotic gut microbiota, alteration of intestinal immunity and systemic inflammation. On these grounds, there are now good prospects for improving the well being of MS patients on the basis of nutritional intervention and healthy life style

2-D zymographic analysis of Broccoli (Brassica oleracea L. var. Italica) florets proteases: Follow up of cysteine protease isotypes in the course of post-harvest senescence.

Zymographic analysis of Broccoli florets (Brassica oleracea L. var. Italica) revealed the presence of acidic metallo-proteases, serine proteases and cysteine proteases. Under conditions which were denaturing for the other proteases, the study was restricted to cysteine proteases. 2-D zymography, a technique that combines IEF and zymography was used to show the presence of 11 different cysteine protease spots with molecular mass of 44 and 47–48 kDa and pIs ranging between 4.1 and 4.7. pI differences could be ascribed to different degrees of phosphorylation that partly disappeared in the presence of alkaline phosphatase. Post-harvest senescence of Broccoli florets was characterized by decrease in protein and chlorophyll contents and increase of protease activity. In particular, as determined by 2-D zymography, the presence of cysteine protease clearly increased during senescence, a finding that may represent a useful tool for the control of the aging process

Unveiling a hidden biomarker of inflammation and tumor progression: The 65 kDa isoform of MMP-9 new horizons for therapy

Cancer metastasis is a stage of the disease where therapy is mostly ineffective; hence, the need to find reliable markers of its onset. The metalloproteinase-9 (MMP-9, gelatinase B) in its 82 kDa active form, is a good candidate, but here we show that the correspondent little known 65 kDa active MMP-9 isoform, often misrepresented with the other gelatinase MMP-2, is a more suitable marker. Sera from patients with lung and breast cancer were analyzed by bidimensional zymography to detect the activity of MMP-9 and MMP-2. Enzyme identity was confirmed by comparison with MMP-9 standards and by western blotting. The 65 kDa isoform of MMP-9 is a suitable biomarker to monitor tumor progression from tissue neoplasms to metastatic stage, as its activity begins to appear when disease severity increases and becomes very high in metastasis. Moreover, the 65 kDa MMP-9, which derives from the 82 kDa MMP-9, no longer responds to natural MMP-9 inhibitors. As its activity cannot be controlled, its appearance may warn that the pathological process is becoming irreversible. Identification and inhibition of the enzymes converting the inhibitor-sensitive 82 kDa MMP-9 into the corresponding “wild” 65 kDa MMP-9 may allow to develop therapies capable of blocking metastases

Efficient recovery of whole cell proteins in Oenococcus oeni - a comparison of different extraction protocols for high-throughput malolactic starter applications

In this study, we compared different total protein extraction protocols to achieve highly efficient isolation and purification of total proteins for the specific protein profiling of Oenococcus oeni. The sodium dodecyl sulfatepolyacrylamide gel electrophoresis patterns obtained for the different extraction protocols revealed not only a qualitative similar protein pattern but also quantitative variations with different intensity bands depending on the extraction method used. The selected extraction method added with sonication proved to work extremely well and efficiently and was able to obtain a high resolution 2- D electrophoresis (2-DE) map. Prominent spots were successfully identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry and corresponded to 76 different proteins involved in the main metabolic pathways. The approach allowed to achieve a protein profiling specific for O. oeni from Aglianico wine with numerous characterized protein products corresponding to many different O. oeni genes and associated with main cellular pathways. Further investigations of the 2-DE protein expression profile will provide useful and interesting information on the molecular mechanisms at the protein level responsible for growth and survival of O. oeni in wine

Total phenols and flavonoids content, antioxidant capacity and lipase inhibition of root and leaf horseradish (Armoracia rusticana) extracts

Horseradish (Armoracia rusticana Gaertn) is a perennial crop belonging to the Brassicaceae family, widely used as spice in foods and herbal ingredient in ethno-medicine. In this study, were evaluated the phenolic compounds content, antioxidant capacity and anti-lipase activity of methanol, methanol/water (70/30, v/v) and methanol/water (50/50, v/v) extracts of horseradish roots and leaves. Among the extracts tested, both roots and leaves aqueous methanolic (70/30, v/v and 50/50, v/v) extracts showed higher total phenol and flavonoid contents and antioxidant capacity than the corresponding methanol extracts. But extraction yield was high for methanol/water (50/50, v/v) extracts, in both roots and leaves. The extracts exhibited anti-lipase activity in dose-dependent manner. The results showed that the extraction yield and the antioxidant capacity were strictly dependent on the solvent polarity. The results suggest that A. rusticana could provide opportunities for the development of functional food and further in vivo studies for obesity treatment

Edible Mushrooms as Source of Fibrin(ogen)olytic Enzymes: Comparison between Four Cultivated Species

Cardiovascular diseases represent the main cause of death. A common feature of cardiovascular disease is thrombosis resulting from intravascular accumulation of fibrin. In the last years, several fibrinolytic enzymes have been discovered in many medicinal or edible mushrooms as potential new antithrombotic agents. This study aimed to compare the fibrin(ogen)olytic activity of crude extracts from the fruiting bodies of four cultivated edible mushrooms: Lentinula edodes, Pleurotus ostreatus, Pleurotus eryngii, and Agrocybe aegerita. Fibrin(ogen)olytic activity was assessed by fibrin plate, spectrophotometric assay and electrophoretic analysis (SDS-PAGE and zymography). The highest activity was detected for P. ostreatus followed by P. eryngii, L. edodes and A. aegerita. Results indicated that enzymes exhibited maximum activity at pH 6–7 and 30–40 C, respectively. Enzyme activity was inhibited by serine and metalloprotease inhibitors. We proposed a new index called the Specific Fibrin(ogen)olytic Index (SFI), which allows specification of the proportion of the total proteolytic capacity due to the fibrin(ogen)olytic activity. These data suggest that the extracts from fruiting bodies or powdered mushrooms can be used as functional ingredients for the development of new functional foods that may act as thrombolytic agents responding, at the same time, to the increasing demand for safe, healthy and sustainable food

May Diet and Dietary Supplements Improve the Wellness of Multiple Sclerosis Patients? A Molecular Approach

Multiple sclerosis is a complex and multifactorial neurological disease, and nutrition is one of the environmental factors possibly involved in its pathogenesis. At present, the role of nutrition is unclear, and MS therapy is not associated to a particular diet. MS clinical trials based on specific diets or dietary supplements are very few and in some cases controversial. To understand how diet can influence the course of MS and improve the wellness of MS patients, it is necessary to identify the dietary molecules, their targets and the molecular mechanisms involved in the control of the disease. The aim of this paper is to provide a molecular basis for the nutritional intervention in MS by evaluating at molecular level the effect of dietary molecules on the inflammatory and autoimmune processes involved in the disease

Antioxidant Activity of Polysaccharides from the Edible Mushroom Pleurotus eryngii

In this study the antioxidant and neuroprotective activity of an enriched polysaccharide fraction (EPF) obtained from the fruiting body of cultivated P. eryngii was evaluated. Proximate composition (moisture, proteins, fat, carbohydrates and ash) was determined using the AOAC procedures. The EPF was extracted by using, in sequence, hot water and alkaline extractions followed by deproteinization and precipitation with cold ethanol. Total - and -glucans were quantified using the Megazyme International Kit. The results showed that this procedure allows a high yield of polysaccharides with a higher content of (1-3; 1-6)- -D-glucans. The antioxidant activity of EPF was detected from the total reducing power, DPPH, superoxide, hydroxyl and nitric oxide radical scavenging activities. The EPF was found to scavenge DPPH, superoxide, hydroxyl and nitric oxide radicals with a IC50 values of 0.52 0.02, 1.15 0.09, 0.89 0.04 and 2.83 0.16 mg/mL, respectively. As assessed by the MTT assay, the EPF was biocompatible for DI-TNC1 cells in the range of 0.006–1 mg/mL and, at concentrations ranging from 0.05 to 0.2 mg/mL, significantly counteracted H2O2-induced reactive oxygen species production. This study demonstrated that polysaccharides extracted from P. eryngii might be used as functional food to potentiate the antioxidant defenses and to reduce oxidative stress

Muscari comosum L. Bulb Extracts Modulate Oxidative Stress and Redox Signaling in HepG2 Cells.

Muscari comosum L. bulbs are commonly used as food in South Italy and also in folk medicine. By evaluating in vitro antioxidant activity and biological activities of their aqueous and methanol extracts, we shed light on the potential role, including both the nutraceutical and health benefits, of this plant. Total polyphenol content (TPC) and total flavonoid content (TFC) were evaluated by the Folin-Ciocalteu method and by the aluminum chloride method, respectively. Antioxidant activity was investigated by three in vitro assays and relative antioxidant capacity index (RACI) was calculated to compare results obtained by different tests. The extracts were tested to evaluate their possible involvement in redox homeostasis, using the human hepatoma (HepG2) cell line used as model. The extracts exhibited concentration/solvent dependent radical scavenging activity, as well as dysregulation of some genes involved in redox pathways by promoting Nrf2, SOD-2, GPX1, ABCC6 and ABCG2 expression. NMR metabolomics analysis suggests that HepG2 cells treated with Muscari comosum extracts experience changes in some metabolites involved in various metabolic pathways

effect of boron and zinc application on hxk1 and makr6 gene expression in strawberry

Boron (B) and zinc (Zn) are important microelements for normal plant activity; their effect have long been studied in numerous agronomical and molecular researches, whereas the interaction of B-Zn remains poorly understood at genomic and transcriptomic levels. Strawberry (fragaria x ananassa) plants were analyzed after single B (80 g/Hl of boron ethanolamine) or Zn (40 g/Hl of Zn-EDTA), and combined B+Zn (80 g/Hl of boron ethanolamine + 40 g/Hl of Zn-EDTA) foliar applications. Leaves, white and red fruits of two genotypes (Candonga and PZ600F13P2 breeding line) were studied. The microelement effects were observed in two conditions: short-term (1 day after treatment) and long-term (20 days after treatment). The differential transcriptomic profiles were analyzed applying RAPD-PCR method. Two genes, HXK1 and MAKR6 have been recognized in differently expressed sequenced amplicons and their expression level was measured using quantitative Real-Time PCR. Fertilizing had an influence on studied genes, their expression differed in relation to the genotype, time course and kind of treatment. Reaction of HXK1 and MAKR6 varied regarding the singular microelement treatments in front of Control (without any treatment) as well as relatively to the combined fertilizing, even if often the zinc effect was "masked" by boron influence in B+Zn treatment. The present study confirmed an initial hypothesis of a significant effects of B and Z in strawberry fruit development. The obtained results, even if they increase the knowledge about the B and Zn effects, could be considered as a first step to details this physiological mechanisms on strawberry fruit development