High molecular weight glutenin subunits in some durum wheat cultivars investigated by means of mass spectrometric techniques

The primary structures of high molecular weight glutenin subunits (HMW-GS) of 5 Triticum durum Desf. cultivars (Simeto, Svevo, Duilio, Bronte, and Sant'Agata), largely cultivated in the south of Italy, and of 13 populations of the old spring Sicilian durum wheat landrace Timilia (Triticum durum Desf.) (accession nos. 1, 2, 3, 4, 7, 8, 9, 13, 14, 15, SG1, SG2, and SG3) were investigated using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and reversed-phase high performance liquid chromatography/nanoelectrospray ionization mass spectrometry (RP-HPLC/nESI-MS/MS). M(r) of the intact proteins determined by MALDI mass spectrometry showed that all the 13 populations of Timilia contained the same two HMW-GS with 75.2 kDa and 86.4 kDa, whereas the other durum wheat cultivars showed the presence of the expected HMW-GS 1By8 and 1Bx7 at 75.1 kDa and 83.1 kDa, respectively. By MALDI mass spectrometry of the tryptic digestion peptides of the isolated HMW-GS of Timilia, the 1Bx and 1By subunits were identified as the NCBInr Acc. No AAQ93629, and AAQ93633, respectively. Sequence verification for HMW-GS 1Bx and 1By both in Simeto and Timilia was obtained by MALDI mass mapping and HPLC/nESI-MSMS of the tryptic peptides. The Bx subunit of Timila presents a sequence similarity of 96% with respect to Simeto, with differences in the insertion of 3 peptides of 5, 9, and 15 amino acids, for a total insertion of 29 amino acids and 25 amino acid substitutions. These differences in the amino acidic sequence account for the determined Δm of 3294 Da between the M(r) of the 1Bx subunits in Timilia and Simeto. Sequence alignment between the two By subunits shows 10 amino acid substitutions and is consistent with the Δm of 148 Da found in the MALDI mass spectra of the intact subunits.     

Oxidative damage mediated by herbicides on yeast cells

Agricultural herbicides are among the most commonly used pesticides worldwide, posing serious concerns for both humans, exposed to these chemicals through many routes, and the environment. To clarify the effects of three herbicides as commercial formulations (namely, Pointer, Silglif, and Proper Energy), parameters related to oxidative issues were investigated on an autochthonous wine yeast strain. It was demonstrated that herbicides were able to affect the enzymatic activities of catalase and superoxide dismutase, as well as to induce carbonylation and thiol oxidation as post-translational modifications of proteins. Saccharomyces cerevisiae is an optimal model system to study responses to xenobiotics and oxidative stress. Thus, the results obtained could further the understanding of mechanisms underlying the toxicity of herbicides.     

A new sustainable formulated feed based on discards from food industries for rearing the sea urchin Paracentrotus lividus (Lmk)

During a 18‐week experiment in a sea‐based system, Paracentrotus lividus was fed with two formulations of a new sustainable feed whose main ingredients were food farming discards, mostly outermost lettuce leaves, in different percentage. Egg white was added as protein source and binder, and a little amount of commercial fish feed was added as lipid source. At the end of the experiment, a high survival rate (>80%) and an increase in test diameter (22%), total weight (130%), gonad weight (240%) and gonad index expressed as size‐adjusted gonad weight (288%) were recorded. Gonads achieved a good colour and high levels of essential and polyunsaturated fatty acids, hence representing a commercially valuable product. Comparing the two feed formulations, the one with the lower lettuce content (57% vs. 67%) led to the best product quality, in terms of gonad features (gonad index and colour) and content of essential fatty acids. In order to move towards a higher sustainability of the aquaculture sector, this study proposes a low‐cost feed, produced with cheap and sustainable ingredients such as food farming discards.     

New Drugs from Marine Organisms in Alzheimer’s Disease

Alzheimer’s disease (AD) is a multifactorial neurodegenerative disorder. Current approved drugs may only ameliorate symptoms in a restricted number of patients and for a restricted period of time. Currently, there is a translational research challenge into identifying the new effective drugs and their respective new therapeutic targets in AD and other neurodegenerative disorders. In this review, selected examples of marine-derived compounds in neurodegeneration, specifically in AD field are reported. The emphasis has been done on compounds and their possible relevant biological activities. The proposed drug development paradigm and current hypotheses should be accurately investigated in the future of AD therapy directions although taking into account successful examples of such approach represented by Cytarabine, Trabectedin, Eribulin and Ziconotide. We review a complexity of the translational research for such a development of new therapies for AD. Bryostatin is a prominent candidate for the therapy of AD and other types of dementia in humans.     

The expression pattern of TIR8 is conserved among vertebrates

IL-1R/TLRs play an important role in the inflammatory responses. Their signaling pathway is tightly regulated to keep inflammation under control. The regulation involves both extracellular and intracellular mechanisms. TIR8, also known as SIGIRR (Single Immunoglobulin IL-1R-Related molecule), is an orphan receptor belonging to the IL-1R/TLRs super-family. Recent studies suggest its role in modulating the inflammatory response in the gastrointestinal tract in mice, acting as an IL-1 receptor family antagonist. We identified the sequence and analyzed the expression of TIR8 in a wide panel of organs and tissues in different animals. There was high homology of the cDNA sequence among human, mouse and the domestic species (74–85%). The pattern of expression of this receptor was similar in all the species examined (high levels in kidney and gastrointestinal tract) and similar to the human. These results demonstrate that TIR8 is conserved, in evolutionary terms, both with regard to sequence and pattern of expression, a finding consistent with a key function of this molecule in modulating inflammation in the gastrointestinal tract.     

Flavonoid-induced autophagy in hormone sensitive breast cancer cells

The activity of 8-prenylapigenin (8-PA) and its 3'-methoxylated analogue isocannflavin B (IsoB) was investigated in estrogen-dependent T47-D and estrogen-independent MDA-MB-231 human breast cancer cell lines. 8-PA showed a biphasic effect on T47-D cell proliferation, while no significant effect was observed on MDA-MB-231 cells. Conversely, IsoB exhibited only an inhibitory effect on T47-D cell proliferation, accompanied by the appearance of an intense intracytoplasmic vacuolization of autophagic origin. Moreover, biochemical analysis showed that IsoB reduced Akt phosphorylation and p21^Cip1 expression in T47-D cells. These data show that the prenylflavone moiety is a versatile platform for the induction and modulation of bioactivity.     

Wettability influences cell behavior on superhydrophobic surfaces with different topographies

Surface wettability and topography are recognized as critical factors influencing cell behavior on biomaterials. So far only few works have reported cell responses on surfaces exhibiting extreme wettability in combination with surface topography. The goal of this work is to study whether cell behavior on superhydrophobic surfaces is influenced by surface topography and polymer type. Biomimetic superhydrophobic rough surfaces of polystyrene and poly(L-lactic acid) with different micro/nanotopographies were obtained from smooth surfaces using a simple phase-separation based method. Total protein was quantified and showed a less adsorption of bovine serum albumin onto rough surfaces as compared to smooth surfaces of the same material. The mouse osteoblastic MC3T3-E1 cell line and primary bovine articular chondrocytes were used to study cell attachment and proliferation. Cells attached and proliferate better in the smooth surfaces. The superhydrophobic surfaces allowed cells to adhere but inhibited their proliferation. This study indicates that surface wettability, rather than polymer type or the topography of the superhydrophobic surfaces, is a critical factor in determining cell behavior.     

Characterisation of the first <Emphasis Type="Italic">Stemphylium vesicarium</Emphasis> isolates resistant to strobilurins in Italian pear orchards

Up to 2005 the sensitivity of Stemphylium vesicarium (Wallr.) Simm., the causal agent of pear brown spot, to the strobilurin fungicides kresoxim-methyl, trifloxystrobin and pyraclostrobin was still comparable with baseline values associated with good efficacy in the field. During 2006, the first resistant isolates were detected in two commercial pear orchards in the Emilia-Romagna region (Italy), one of which was affected by considerable control failure linked to strobilurin treatments as demonstrated in a field trial. In vitro sensitivity tests with 0.5 mg l⁻¹ of kresoxim-methyl, trifloxystrobin and pyraclostrobin showed that in the population collected in the orchard with control failure the conidial germination was greater than 90% compared to an untreated control both in 2006 and in 2007, i.e. 1 year after the suspension of strobilurin applications. In the other orchard, where only a few symptomatic fruits were found and the strobilurins were still in use, the conidial germination was lower, about 50% in 2006 and 25% in 2007. The molecular analysis of mitochondrial cytochrome b gene of some monospore isolates with different levels of sensitivity confirmed the presence of the mutation causing G143A substitution in all the resistant isolates. In conclusion, both in vitro tests and molecular analysis confirmed the first occurrence of Stemphylium vesicarium resistance to all strobilurin fungicides tested.