Phenolic Content and Antioxidant Activity in Raw and Denatured Aqueous Extracts from Sprouts and Wheatgrass of Einkorn and Emmer Obtained under Salinity.

Total phenolic content (TPC), reducing power (RP), superoxide radical scavenging (RS), and thiobarbituric acid reactive substances (TBARS) production inhibition were measured in raw and denatured aqueous extracts from sprouts and wheatgrass of einkorn and emmer obtained at increasing salinity. Grains were incubated and kept at 0, 25, 50, and 100 mM NaCl until either sprout or wheatgrass stage. Additionally, a recovery treatment was included, in which sprouts obtained at 100 mM NaCl were then transferred at 0 mM NaCl until wheatgrass stage. All parameters (TPC, RP, RS, and TBARS production inhibition) increased with sprouting and were highest in wheatgrass. Salinity increased all parameters, but the effect varied with NaCl concentration, genotype, developmental stage, and plant material processing (raw or denatured). Overall, given the delay and limitation of growth at high NaCl concentration, the best compromise appears to be the application of a moderate salinity (25 to 50 mM NaCl). In denatured extracts, TPC, RP, and RS slightly decreased, and TBARS was not affected, which means that antioxidant activity was mainly related to compounds other than enzymes and peptides, and thus it can be assumed to remain after digestion. Thus, supplementing the human diet with einkorn or emmer sprouts and wheatgrass can actually benefit health.

Biochemical and mass spectrometry recognition of phospholipid-peptide complexes in wheat sprouts extract.

Total hydroalcoholic extract of wheat sprouts was treated with 90% cold acetone as a preliminary step directed to separate antioxidant peptides from antioxidant polyphenols. Surprisingly, the addition of acetone causes the formation of a yellow buoyant gelatinous drop that prevailingly contains peptides and phospholipids. In this context, evidences have been presented that support the hypothesis that peptides (and perhaps other active molecules) are complexed with phospholipids. In fact, the MS/MS analysis of some main ions, present in RP HPLC fractions of wheat sprout extract, generates several ions that correspond to molecular weight of phospholipids or phospholipid fragments. Moreover, several ions were detected that correspond to lysophosphatidylcholine or phosphatidylcholine-peptide complexes. The possibility that phospholipids can be complexed with peptides has been discussed in the light of potential involvement in the peptide bioavailability.

Interaction of DDSDEEN peptide with N-CAM protein. Possible mechanism enhancing neuronal differentiation.

DDSDEEN chromatin peptide, after dansylation, was studied for its ability to bind N-CAM protein. The binding causes a quenching of the Dns-peptide fluorescence emission. Dose- and time-dependent binding of Dns-peptide with N-CAM has been shown. Fluorescence quenching is completely lost if the Dns-peptide is subjected to carboxypeptidase digestion. Moreover the undansylated peptide pEDDSDEEN competes with the DnsDDSDEEN peptide for the binding with the N-CAM protein. The Dns-peptide-N-CAM bond has been related to the peptide biological activity probably involved in the promotion of neuronal differentiation. An attempt to recognize a possible N-CAM binding site for Dns-peptide was performed by alignment of N-CAM from various sources with some sequences that have been previously reported as binding sites for the pEDDSDEEN and DDSDEEN peptides. Interestingly, the alignment of N-CAM from various sources with the peptides WHPREGWAL and WFPRWAGQA recognizes on rat and human N-CAM a unique sequence that could be the specific binding site for chromatin peptide: WHSKWYDAK. This sequence is present in fibronectin type-III domain of N-CAM. In addition molecular modeling studies indicate the N-CAM sequence WHSKWYDAK as, probably, the main active site for DnsDDSDEEN (or pEDDSDEEN) peptide ligand. Accordingly the binding experiments show a high affinity between WHSKWYDAK and DnsDDSDEEN peptides.

Short acidic peptides isolated from wheat sprout chromatin and involved in the control of cell proliferation. Characterization by infrared spectroscopy and mass spectrometry.

Low molecular weight peptides were isolated from the chromatin of wheat sprouts. Following gel filtration the peptide fraction shows a sharp inhibiting activity on the growth of HeLa cancer cells. Infrared (IR) spectroscopy and mass spectrometry have been utilized to characterize the wheat sprout peptides in an attempt to recognize the peptide sequence involved in the control of cell growth. The quantitative presence of a peptide with MH+=572 appears proportional to the cell growth inhibition activity. This compound has been subjected to extensive mass spectrometry analysis. The automatic computational analysis of the ions of second, third and fourth generations indicate a peptide sequence, AcHis-Asp-Ser-Glu-, that binds at the C-terminal a molecule of ethanolamine. Moreover, the results show that some sequences of the wheat sprout peptide family are present in the peptide fractions isolated from several other tissues, thus supporting the hypothesis of ubiquitous regulatory peptides.

NPPB block of the intermediate-conductance Ca2+-activated K+ channel.

We have shown that the Cl(-) channel blocker 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) also blocks the intermediate-conductance Ca(2+)-activated K(+) (IK(Ca)) current in human leukemic HL-60 and glioblastoma GL-15 cell lines. The macroscopic IK(Ca) current was activated by ionomycin plus 1-EBIO, and identified as intermediate conductance by being fully blocked by charybdotoxin, clotrimazole, nitrendipine (L-type Ca(2+) channel blocker), and NS1619 (BK(Ca) channel opener), but not by D-tubocurarine or TEA. The IK(Ca) current was blocked by NPPB in a reversible dose-dependent manner, with an IC(50) of 39 microM in HL-60 and 125 microM in GL-15 cells. The block of the IK(Ca) current was also recorded at the single channel level in excised inside-out patches. As expected, NPPB also blocked the volume-activated Cl(-) current expressed by GL-15 cells, with an IC(50) of 44 microM. The functional implications of IK(Ca) current block by NPPB are discussed.

Synthesis of antioxidants in wheat sprouts.

Aqueous and ethanolic extracts from wheat (Triticum aestivum) sprout powder were analyzed to determine its reduction and antioxidant activities. Mean and standard deviation of five independent samples were reported. The results showed that the micromoles of potassium ferricyanide reduced by aqueous and ethanolic extracts corresponding to 1 g of sprout powder (80.6 +/- 11.2 and 9.7 +/- 1.8, respectively) were higher than that reduced by 1 mg of other reducing compounds: ascorbic acid, rutin, quercetin, and reduced gluthatione (4.8 +/- 0.7, 3.8 +/- 1.2, 4.8 +/- 1.7, and 1.6 +/- 0.4, respectively). In addition, the oxygen superoxide scavenging activity performed by sprout extracts (from 1 g of powder) is comparable to that shown by 10 mg of antioxidant pure compounds (rutin and quercetin). Biochemical analysis of the sprout extracts shows that the antioxidant activity is mainly due to reducing glycoside and polyphenolic compounds.

Characterization of phospholipid molecular species and peptide molecules in wheat sprout hydroalcoholic extract.

The phospholipid molecular species and the main peptide molecules of wheat sprout hydroalcoholic extract have been fully characterized by normal-phase high performance liquid chromatography coupled online with positive electrospray ionization tandem mass spectrometry. The extract that resulted was rich in phospholipid molecular species formed by the combination of the two essential fatty acids (α-linoleic and α-linolenic). These species accounted for 51.7% of total phosphatidic acid, 47.3% of total phosphatidylethanolamine, 37.7% of total phosphatidylcholine, and 14.1% of total phosphatidylinositol. The last one showed the highest amounts of species containing palmitic acid, thus representing the most saturated phospholipid class. The extract was also shown to contain several peptide sequences with both potential antioxidant domains and interaction sites for phospholipids (i.e., H-Ala-Gly-Ser-Met-Met-Cys-NH2, H-Tyr-Met-Thr-Val-Val-Ala-Cys-NH2, etc.); this latter finding can have a highly positive impact on the poor peptides bioavailability. Because of the presence of essential fatty acids-rich phospholipids and bioactive peptides, wheat sprout hydroalcoholic extract can be considered a potential functional food ingredient.

Comparative activity of antioxidants from wheat sprouts, Morinda citrifolia, fermented papaya and white tea.

Hydroalcoholic extracts from wheat sprouts, white tea, Morinda citrifolia and fermented papaya were analysed to determine their reducing power and antioxidant activity. The results show that the micromoles of potassium ferricyanide reduced by a quantity of extract corresponding to 1 g of the various dehydrated starting tissues are: 12.91+/-0.83 (wheat sprouts), 10.66+/-1.22 (M. citrifolia), 17.06+/-1.24 (white tea), and 1.05+/-0.09 (fermented papaya). In addition the results show a strong oxygen superoxide scavenging activity in the extracts from white tea, M. citrifolia and wheat sprouts. The activity of the fermented papaya extract is the lowest. The thin-layer chromatography and UV spectrophotometry of the extracts show in each source a mixture of antioxidant compounds probably belonging to the families of reducing glycosides and polyphenols. The chromatographic pattern of the antioxidant compounds and the UV spectrum are quite different in the various sources.

Aging reversibility: from thymus graft to vegetable extract treatment-- application to cure an age-associated pathology.

Neonatal thymus graft and thymus calf extract (TME) in vivo treatment exert similar corrective actions on different mouse age-related alterations. The aim of the present paper is to investigate whether a vegetal extract, wheat sprout extract (WESPRE), could mimic the thymus action on recovering age-related alterations and if this extract can cure an age-associated pathology, the cataract in dogs. Present experiments were carried out by using WESPRE and TME in vivo in old mice to check their ability to recover the altered DNA synthesis in hepatocyte primary cultures. Old mice treated with WESPRE and TME showed a recovery of hepatocyte DNA synthesis levels when compared with the old untreated ones. The increase of DNA and protein contents observed in aged animals is reduced by WESPRE treatments to levels observed in young mice hepatocytes. We measured also WESPRE phosphorylation activity by endogenous kinase: it was from 10 to 40 times higher with respect to wheat seeds. Old dogs were orally treated for a month and the lens opacity analysed before and after the treatment. Results showed a reduction from 25 to 40% of lens opacity. The efficacy of wheat sprouts in the recovery of age-related alterations and in treating age-associated pathologies could be due to the contemporary presence of small regulatory acid peptides, a remarkable level of highly energetic phosphoric radicals and antioxidant molecules, peculiarities that may be, to some extent, related to the aging process regulation.

Nutritional relevance of wheat sprouts containing high levels of organic phosphates and antioxidant compounds.

Wheat sprouts contain a very high level of organic phosphates and a powerful cocktail of antioxidant molecules. Catalase and peroxidase activity appears very strong. Regarding low-molecular weight antioxidant molecules, TLC and other biochemical analysis show the presence of several classes of antioxidant compounds such as reducing glycosides, polyphenols, and -SH groups. Antioxidant activity has been measured by potassium ferricyanide reduction and by means of superoxide scavenging NBT. It has also been observed that the oral assumption of wheat sprout powder induces in old dogs a significant reduction of senile cataract. These results clearly confirm that the administration of natural antioxidant compounds can prevent or delay senile cataract. Moreover, it is evident that wheat sprout biologically active substances can be at least partially absorbed during the digestion process. Our research is now directed to the recovery of large amounts of wheat sprout biologically active substances that could also be used in the formulation of nutritional complements.