Plant poisoning leads to alpha-synucleinopathy and neuromelanopathy in kangaroos.

The pathogenesis of synucleinopathies, common neuropathological lesions normally associated with some human neurodegenerative disorders such as Parkinson's disease, dementia with Lewy bodies and multiple system atrophy, remains poorly understood. In animals, ingestion of the tryptamine-alkaloid-rich phalaris pastures plants causes a disorder called Phalaris staggers, a neurological syndrome reported in kangaroos. The aim of the study was to characterise the clinical and neuropathological changes associated with spontaneous cases of Phalaris staggers in kangaroos. Gross, histological, ultrastructural and Immunohistochemical studies were performed to demonstrate neuronal accumulation of neuromelanin and aggregated α-synuclein. ELISA and mass spectrometry were used to detect serum-borne α-synuclein and tryptamine alkaloids respectively. We report that neurons in the central and enteric nervous systems of affected kangaroos display extensive accumulation of neuromelanin in the perikaryon without affecting neuronal morphology. Ultrastructural studies confirmed the typical structure of neuromelanin. While we demonstrated strong staining of α-synuclein, restricted to neurons, intracytoplasmic Lewy bodies inclusions were not observed. α-synuclein aggregates levels were shown to be lower in sera of the affected kangaroos compared to unaffected herd mate kangaroos. Finally, mass spectrometry failed to detect the alkaloid toxins in the sera derived from the affected kangaroos. Our preliminary findings warrant further investigation of Phalaris staggers in kangaroos, potentially a valuable large animal model for environmentally-acquired toxic synucleinopathy.

Chemical Composition, Characteristics Profiles and Bioactivities of Tunisian Phalaris canariensis Seeds: a Potential Source of ω-6 and ω-9 Fatty Acids.

Seeds oils of Phalaris canariensis extracted by ultrasonication and cold maceration were evaluated for their physical characteristics, total phenol contents, fatty acid and sterol compositions as well as for their antioxidant, antibacterial and acetylcholinesterase activities. The physicochemical properties of ultrasonication and cold maceration oils respectively were: acid values (4.00 and 3.25) mg KOH/g, peroxide values (5.53 and 4.41) meq O2 Kg-1, iodine values (88.83 and 95.17) g/100 g of oil, saponification values (119.21 and 98.17) mg KOH/g, phenolic content (36.40 and 53.00) mg GAE/g extract, chlorophylls (0.52 and 0.60) mg/kg oil and carotenoids contents (1.92 and 1.88) mg/kg oil. Gas chromatography analysis revealed that linoleic (52.03 and 52.2%), oleic (31.75 and 31.84%) and palmitic (11.09 and 11.34 %) acids were the major fatty acids in the two oils. Specific extinctions at 232 nm (K232) and 270 nm (K270) were (0.58 and 0.44) and (0.42 and 0.33), respectively. The DSC melting curve showed that their melting points and melting enthalpies were (-28.05°C and 76.8 J/g) and (-27.47°C and 62.3 J/g), respectively. On the other hand, the evaluation of their DPPH radical scavenging, total antioxidant capacity, antibacterial and acetylcholinesterase activities showed interesting results. Thus, Phalaris canariensis seeds oils could deserve further consideration and investigation as a potentially new multi-purpose product for agro-food, medicinal and cosmetic uses.

Antioxidant and Antihypertensive Potential of Protein Fractions from Flour and Milk Substitutes from Canary Seeds (Phalaris canariensis L.).

Canary seed (Phalaris canariensis) is used to feed birds but it has been recently considered a promising cereal with nutraceutical potential for humans. The aim of this work was to analyze the protein fractions from canary seed flour and from milk substitutes (prepared by soaking the seeds in water 12 and 24 h), and to evaluate antioxidant and antihypertensive capacity of peptides obtained after in vitro digestion. Prolamins were the major protein fraction, followed by glutelins. After digestion, albumins and prolamins fractions from milks presented higher levels of peptides than flour, globulins showed more peptides in flour and glutelins were found in similar concentrations in all samples; 24 h milk prolamins had the highest concentration of peptides. Purification by high performance liquid chromatography (HPLC), sequencing of peptides, in vitro antioxidant ABTS (2,2'-azino-bis, 3-ethylbenzothiazoline-6-sulphonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays, and antihypertensive capacity (angiotensin converting enzyme (ACE) assay), indicated that peptides from canary seed prolamins were the most efficient compounds with antioxidant and antihypertensive activity. Canary seeds may be considered an accessible and cheap source to prepare milk substitutes with high contents of bioactive peptides with remarkable functional properties to promote better human health and healthy ageing.

Inhibition by Seeds of Phalaris canariensis Extracts of Key Enzymes Linked to Obesity.

CONTEXT: Obesity and its associated diseases are an increasing problem around the world. One hyperglycemic remedy is reduction of glucose absorption performed by suppressing digestion of carbohydrates and lipids through the use of inhibitors. Phalaris canariensis (P canariensis) is a species belonging to the Graminaceae family and is used in traditional medicine in Mexico for treatment of diabetes and obesity. OBJECTIVE: The aim of the study was to evaluate the effects of different extracts of the seeds of P canariensis on enzymes metabolizing fat and carbohydrates, obtained using 3 solvents. DESIGN: The seeds of P canariensis were extracted using hexane (ALH), chloroform (ALC), and methanol (ALM) and were investigated for their antiobesity potential. SETTING: This research was conducted in the Laboratory of Research of Natural Products in the School of Chemical Engineering at the National Polytechnic Institute and in the Research Laboratory of Enzymology in the National School of Biological Sciences. OUTCOME MEASURES: Different concentrations of the extracts were used to study the inhibition of enzymatic activity by porcine pancreatic α-amylase, with carbose as a positive control. The inhibitory activity of α-glucosidase was determined using the standard method with bovine serum albumin (BSA). Pancreatic lipase (PL) activity was measured by absorbance at 412 nm, and the data obtained were compared with orlistat. The PL activity was assessed using a second method measuring the rate of release of oleic acid from triolein. Lipoprotein lipase (LPL) activity was measured by released (3H)-oleic acid. Lipolytic activity in cultured, mouse, 3T3-Ll adipocytes was used as a measure of hormone-sensitive lipase activity. The inhibitory activity of rat intestinal sucrase was determined by measuring the glucose released. A Caco-2 cell assay determined the content of free glucose. RESULTS: The ALH extract of P canariensis showed potent inhibitory activity with IC50 values of 2.13 and 1.25 mg/mL as compared with α-amylase and α-glucosidase, respectively, and produced inhibition in rat intestinal sucrose. Further, the ALM extract showed significantly inhibitory effects against PL, LPL, and lipolysis of 3T3-LI adipocytes. CONCLUSIONS: The results provide evidence for the effects of the seeds of P canariensis for a retarded absorption of carbohydrates and lipids through the inhibition of enzymes that are related to obesity and diabetes mellitus type 2.

Evaluation of Anti-inflammatory Activity of Seeds of Phalaris canariensis.

Chloroform extract (ALC) from the seeds of Phalaris canariensis were assayed for antiinflammatory activity by carrageenan-induced oedema, cotton pellets-induced granuloma, histamine-induced inflammation, croton oil-induced oedema, activity of myeloperoxidase (MPO), adjuvant-induced arthritis, quantification of TNFα, IL-1ß, PGE2 and LTB4 and nitric oxide (NO) assay. ALC exhibited significant anti-inflammatory activity in different chemically-induced edemas in a dose dependent manner. In the chronic model cotton pellets-induced granuloma showed decreased formation of granuloma tissue. Also caused inhibition of ear inflammation edema and influx of polymorphonuclear cells, as evidence by a decrease in ear thickness and reduced myeloperoxidase activity and inhibit mediators of inflammation as TNFα, IL-1ß, PGE2 and LTB4. When RAW 264.7 macrophages were treated with ALC together with LPS a significant inhibition of NO production was detected. These data provide evidence for antiinflammatory effect of P. canariensis by mechanisms that involve a reduced neutrophil influx and decreased production of inflammatory cytokines.

Changes of phenolic profiles and antioxidant activity in canaryseed (Phalaris canariensis L.) during germination.

Canaryseed is an important cereal crop in western Canada. The changes of the total phenolic content (TPC), antioxidant activities, phenolic acid profiles (free and bound) of canaryseed during germination were investigated in the present study. The growth properties also were investigated. Fresh weight, shoot length and root length increased, whereas dry mass of canaryseed decreased during germination. A 22.3% loss of dry matter was observed at 120h of germination. The total phenolic content and antioxidant activities of free and bound extracts showed a general trend of germinated seeds>raw seeds>soaked seeds. Free, bound and total phenolic content significantly increased 1042%, 120% and 741% at the end of germination as compared to raw seeds (p<0.05). DPPH, ABTS and ORAC assays were employed to evaluate the antioxidant activity of canaryseed. There were high correlations between total phenolic content and antioxidant activities. TPC and ORAC values showed the highest correlation (r=0.9984). Six phenolic acids in free phenolic extracts and seven phenolic acids in bound phenolic extracts were detected, respectively. Bound ferulic acid, the dominant phenolic acid in canaryseed, significantly increased during germination (p<0.05). Study showed that germination provided a new approach to further develop canaryseed as a functional food for human consumption.

Accumulation and distribution of macroelements in the organs of Phalaris arundinacea L.: Implication for phytoremediation.

The aim of this study was to assess nutrient and alkali metal accumulation and their distribution in the organs of Phalaris arundinacea and relations between environmental macroelement concentrations and accumulation in plant tissues. The content of N, P, K, Ca, Mg and Na in water, bottom sediments and different organs of Phalaris arundinacea from the Bystrzyca River (Lower Silesia) was determined. The organs of the reed canary grass contained relatively high amounts of macroelements and differed significantly in their accumulation. All macroelements other than Na were accumulated in the highest amounts in aboveground, photosynthetic tissues. Phalaris arundinacea is an Na and Ca excluder plant and an N, P, Mg and K accumulator. Transport efficiency from bottom sediments to plant roots was higher than between plant organs. Nitrogen, P and K are taken up actively while Ca passively. The high translocation ratio of nutrients, particularly for Ca, Mg, K and N, makes the reed canary grass suitable for nutrient phytoextraction from water and bottom sediments of eutrophic lakes and rivers. Bottom sediments can be considered the primary source of Ca for Phalaris arundinacea.

Blood pressure reducing effects of Phalaris canariensis in normotensive and spontaneously hypertensive rats.

The birdseed Phalaris canariensis (Pc) is popularly used as an antihypertensive agent. The aqueous extract of Pc (AEPc) was administered in adult normotensive Wistar rats and spontaneously hypertensive rats (SHR) and in prehypertensive young SHR (SHR(Y), 3 weeks old). Animals received AEPc (400 mg·kg(-1)·day(-1), by gavage) for 30 days, then groups were divided into 2 subgroups: one was treated for another 30 days and the other received water instead of AEPc for 30 days. AEPc reduced systolic blood pressure (SBP) in both adult groups; however, treatment interruption was followed by a gradual return of the SBP to baseline levels. SHR(Y) became hypertensive 30 days after weaning. AEPc minimized the increase in SBP in SHR(Y), but blood pressure rose to levels similar to those in the untreated group with treatment interruption. There were no changes in renal function, diuresis, or Na(+) excretion. Pc is rich in tryptophan, and the inhibition of the metabolism of tryptophan to kynurenine, a potential vasodilator factor, prevented the blood pressure reducing effect of AEPc. Moreover, AEPc significantly reduced sympathoexcitation. Data indicate that the metabolic derivative of tryptophan, kynurenine, may be a mediator of the volume-independent antihypertensive effect of Pc, which was at least in part mediated by suppression of the sympathetic tonus.

Fractionation of hairless canary seed (Phalaris canariensis) into starch, protein, and oil.

Canary seed is an important specialty crop in Canada. The current market for this true cereal (i.e., belonging to the family Poaceae as wheat) is limited to feed for caged birds. However, canary seed holds a promise for many food and industrial applications based on its composition. Three wet milling procedures based on ethanol (E), water (W), and alkaline (A) extractions used in different order were investigated to determine extraction efficiency and purity of starch, protein, oil, and fiber separated from hairless canary seed, a variety developed for human consumption. Highest extraction efficiencies were obtained when canary seed was defatted with ethanol and then extracted with alkali and water (EAW process). Using this process, approximately 92% pure starch, 75% pure protein, and oil were recovered from canary seed groats. The highest purity of protein, however, was obtained when canary seed was fractionated by the EWA process, that is, defatted and then extracted with water followed by alkali. Fiber component separated prior to alkaline extraction contained high amounts of nonfiber components as indicated by its yield. The EAW extraction process seems to be more promising in canary seed fractionation based on recovery and purity of components.

Selenium concentrations of common weeds and agricultural crops grown in the seleniferous soils of northwestern India.

The plants grown in seleniferous soils constitute a major source of toxic selenium levels in the food chain of animals and human beings. Greenhouse and field experiments were conducted to study selenium concentrations of weeds, forages and cereals grown on seleniferous soils located between 31.0417 degrees to 31.2175 degrees N and 76.1363 degrees to 76.4147 degrees E in northwestern India. Eleven winter season (November-April) weed plants were grown in the greenhouse in a soil treated with different levels of selenate-Se. Selenium concentrations of weed plants increased progressively with the levels of selenate-Se in soil. The highest Se concentration was recorded by Silene gallica (246 mgkg(-1)) and the lowest by Avena ludoviciana (47 mgkg(-1)) at 2.5 mg Sekg(-1) soil. A.ludoviciana and Spergula arvensis proved highly tolerant to the presence of 1.25 and 2.5 mg selenate-Sekg(-1) soil and the remaining weeds were sensitive to Se. Dry matter yield of Se-sensitive weed plants was 25 to 62% of the yield in the no-Se control at 1.25mg selenate-Sekg(-1) and 6 to 40% at 2.5mg selenate-Sekg(-1) soil. Other symptoms like change in leaf colour and size, burning of leaf tips and margins, and delayed flowering were also observed due to Se. Dry matter yield of Se-sensitive weed plants expressed as percentage of yield in the no-Se control at both the Se levels was inversely correlated with their Se content (r=-0.731, p<0.01, N=17). Among the weed plants grown in seleniferous soils under field situations, Mentha longifolia accumulated the highest Se (365 mgkg(-1)) and Phalaris minor the lowest (34 mgkg(-1)). Among agricultural crops grown on a naturally contaminated soil in the greenhouse, Se concentrations were the highest for oilseed crops (19-29 mgkg(-1)), followed by legumes (6-13 mgkg(-1)) and cereals (2-18 mgkg(-1)). Helianthus annuus among the oilseed crops, A.ludoviciana among the winter season weeds, M.longifolia among the summer season (May-October) weeds and Cirsium arvense among the perennial weeds can be used for phytoremediation of seleniferous soils as these accumulate the highest amounts of Se.

Metals and arsenic in soils and corresponding vegetation at Central Elbe river floodplains (Germany).

Floodplain soils at the Elbe river are frequently polluted with metals and arsenic. High contents of these pollutants were detected down to subsoil layers. NH4NO3-extractable (phytoavailable) Cd, Ni, and Zn were elevated in horizons with high acidity. Among five common floodplain plant species, Artemisia vulgaris showed highest concentrations of Cd, Cu, and Hg, Alopecurus pratensis of As and Phalaris arundinacea of Ni, Pb, and Zn. Relationships were weak between metal concentrations in plants and phytoavailable stocks in soil. As and Hg uptake seems to be enhanced on long submerged soils. Enrichments of Cd and Hg are linked to a special plant community composition. Grassland herbage sampled in July/August revealed higher concentrations of As (+122%), Hg (+124%), and Pb (+3723%) than in May. To limit harmful transfers into the food chain, low-lying terraces and flood channels revealing highest contaminations or phytoavailabilities should be excluded from mowing and grazing.

Efficient and sensitive method for quantitative analysis of alkaloids in hardinggrass (Phalaris aquatica L.).

An efficient high-performance thin-layer chromatography (HPTLC) method for the analysis of alkaloids in hardinggrass (Phalaris aquatica L.) was developed. The method employed HPTLC glass plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of ethyl acetate/chloroform/7 N NH4OH in methanol (8:2:1, v/v/v). Using unidimensional double-development, bands were well separated for 10 alkaloid standards as well as alkaloids observed in hardinggrass plant extracts. Identities of compounds observed using HPTLC were validated by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Software was used to quantify individual alkaloids in plant samples based on HPTLC retention factors and intensities relative to standards of known concentration. Correlation coefficients of 0.99 were obtained between estimated and actual concentrations for four standards (methyltyramine, hordenine, gramine, and 5-methoxydimethyltryptamine), with linearity in the range of 120-3840 ng/spot. The HPTLC method is repeatable and specific for beta-carboline, tryptamine, gramine, and tyramine type alkaloids in mixed standard and plant extracts. Initial results indicate substantial variation in alkaloid composition among and within hardinggrass populations.