Phytochemicals of apple peels: isolation, structure elucidation, and their antiproliferative and antioxidant activities

Bioactivity-guided fractionation of Red Delicious apple peels was used to determine the chemical identity of bioactive constituents, which showed potent antiproliferative and antioxidant activities. Twenty-nine compounds, including triterpenoids, flavonoids, organic acids and plant sterols, were isolated using gradient solvent fractionation, Diaion HP-20, silica gel, and ODS columns, and preparative HPLC. Their chemical structures were identified using HR-MS and 1D and 2D NMR. Antiproliferative activities of isolated pure compounds against HepG2 human liver cancer cells and MCF-7 human breast cancer cells were evaluated. On the basis of the yields of isolated flavonoids (compounds 18- 23), the major flavonoids in apple peels are quercetin-3-O-beta-D-glucopyranoside (compound 20, 82.6%), then quercetin-3-O-beta-D-galactopyranoside (compound 19, 17.1%), followed by trace amounts of quercetin (compound 18, 0.2%), (-)-catechin (compound 22), (-)-epicatechin (compound 23), and quercetin-3-O-alpha-L-arabinofuranoside (compound 21). Among the compounds isolated, quercetin (18) and quercetin-3-O-beta-D-glucopyranoside (20) showed potent antiproliferative activities against HepG2 and MCF-7 cells, with EC 50 values of 40.9 +/- 1.1 and 49.2 +/- 4.9 microM to HepG2 cells and 137.5 +/- 2.6 and 23.9 +/- 3.9 microM to MCF-7 cells, respectively. Six flavonoids (18-23) and three phenolic compounds (10, 11, and 14) showed potent antioxidant activities. Caffeic acid (10), quercetin (18), and quercetin-3-O-beta-D-arabinofuranoside (21) showed higher antioxidant activity, with EC 50 values of <10 microM. Most tested flavonoids and phenolic compounds had high antioxidant activity when compared to ascorbic acid and might be responsible for the antioxidant activities of apples. These results showed apple peel phytochemicals have potent antioxidant and antiproliferative activities.     

Unearthing the role of biological diversity in soil health

The soil provides a great variety of microhabitats for myriad organisms of different size, physiological activity, behavior and ecosystem function. Besides abundance of participating soil organisms, their species diversity facilitates maximum exploitation of the resources available in the different habitats. At various levels of resolution, species can be categorized into classes performing ecosystem functions and, within each functional class, into guilds of species with similar life course characteristics. Measurement of the diversity and abundance of species within a functional class provides insights into the nature of ecosystem functions and services and to the health of the soil. At higher resolution, species diversity within guilds of a functional class may infer the degree of exploitation of available resources and the complementarity of an ecosystem service; diversity among the guilds of a functional class may indicate successional complementarity of the services. A diversity of guilds within a functional class expands the range of conditions over which ecosystem services are performed while species diversity within a functional class and its guilds contributes to the magnitude of the services. Consequently, diversity of species within functional classes is a key element of the biological component of soil health. In the context of ecosystem services and soil health, the biomass or metabolic activity of species are more useful measures of their abundance than numbers of individuals. Thus, understanding of soil health and ecosystem function requires, besides knowledge of species diversity within functional classes, assessment of the range of functions currently performed in the system and the abundances of organisms by which they are performed. We propose a diversity-weighted abundance product for comparison of the functional magnitude of different assemblages of like organisms.     

Phytochemicals of black bean seed coats: isolation, structure elucidation, and their antiproliferative and antioxidative activities

Bioactivity-guided fractionation of black bean (Phaseolus vulgaris) seed coats was used to determine the chemical identity of bioactive constituents, which showed potent antiproliferative and antioxidative activities. Twenty-four compounds including 12 triterpenoids, 7 flavonoids, and 5 other phytochemicals were isolated using gradient solvent fractionation, silica gel and ODS columns, and semipreparative and preparative HPLC. Their chemical structures were identified using MS, NMR, and X-ray diffraction analysis. Antiproliferative activities of isolated compounds against Caco-2 human colon cancer cells, HepG2 human liver cancer cells, and MCF-7 human breast cancer cells were evaluated. Among the compounds isolated, compounds 1, 2, 6, 7, 8, 13, 14, 15, 16, 19, and 20 showed potent inhibitory activities against the proliferation of HepG2 cells, with EC50 values of 238.8 +/- 19.2, 120.6 +/- 7.3, 94.4 +/- 3.4, 98.9 +/- 3.3, 32.1 +/- 6.3, 306.4 +/- 131.3, 156.9 +/- 11.8, 410.3 +/- 17.4, 435.9 +/- 47.7, 202.3 +/- 42.9, and 779.3 +/- 37.4 microM, respectively. Compounds 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 14, 15, 19, and 20 showed potent antiproliferative activities against Caco-2 cell growth, with EC50 values of 179.9 +/- 16.9, 128.8 +/- 11.6, 197.8 +/- 4.2, 105.9 +/- 4.7, 13.9 +/- 2.8, 35.1 +/- 2.9, 31.2 +/- 0.5, 71.1 +/- 11.9, 40.8 +/- 4.1, 55.7 +/- 8.1, 299.8 +/- 17.3, 533.3 +/- 126.0, 291.2 +/- 1.0, and 717.2 +/- 104.8 microM, respectively. Compounds 5, 7, 8, 9, 11, 19, 20 showed potent antiproliferative activities against MCF-7 cell growth in a dose-dependent manner, with EC50 values of 129.4 +/- 9.0, 79.5 +/- 1.0, 140.1 +/- 31.8, 119.0 +/- 7.2, 84.6 +/- 1.7, 186.6 +/- 21.1, and 1308 +/- 69.9 microM, respectively. Six flavonoids (compounds 14-19) showed potent antioxidant activity. These results showed the phytochemical extracts of black bean seed coats have potent antioxidant and antiproliferative activities.     

Phytochemicals and antioxidant activity of milled fractions of different wheat varieties

The health-promoting effects of whole-grain consumption have been attributed in part to their unique phytochemical contents and profiles that complement those found in fruits and vegetables. Wheat is an important component of the human diet; however, little is known about the phytochemical profiles and total antioxidant activities of milled fractions of different wheat varieties. The objectives of this study were to investigate the distribution of phytochemicals (total phenolics, flavonoids, ferulic acid, and carotenoids) and to determine hydrophilic and lipophilic antioxidant activity in milled fractions (endosperm and bran/germ) of three different wheat varieties, two of which were grown in two environments. Grain samples of each of the wheat varieties were milled into endosperm and bran/germ fractions. Each fraction was extracted and analyzed for total phenolics, ferulic acid, flavonoids, carotenoid contents, and hydrophilic and lipophilic antioxidant activities. Total phenolic content of bran/germ fractions (2867-3120 micromol of gallic acid equiv/100 g) was 15-18-fold higher (p < 0.01) than that of respective endosperm fractions. Ferulic acid content ranged from 1005 to 1130 micromol/100 g in bran/germ fractions and from 15 to 21 micromol/100 g in the endosperm fractions. The bran/germ fraction flavonoid content was 740-940 micromol of catechin equiv/100 g. On average, bran/germ fractions of wheat had 4-fold more lutein, 12-fold more zeaxanthin, and 2-fold more beta-cryptoxanthin than the endosperm fractions. Hydrophilic antioxidant activity of bran/germ samples (7.1-16.4 micromol of vitamin C equiv/g) was 13-27-fold higher than that of the respective endosperm samples. Similarly, lipophilic antioxidant activity was 28-89-fold higher in the bran/germ fractions (1785-4669 nmol of vitamin E equiv/g). Hydrophilic antioxidant activity contribution to the total antioxidant activity (hydrophilic + lipophilic) was >80%. In whole-wheat flour, the bran/germ fraction contributed 83% of the total phenolic content, 79% of the total flavonoid content, 51% of the total lutein, 78% of the total zeaxanthin, 42% of the total beta-cryptoxanthin, 85% of the total hydrophilic antioxidant activity, and 94% of the total lipophilic antioxidant activity. Our results showed that different milled fractions of wheat have different profiles of both hydrophilic and lipophilic phytochemicals. These findings provide information necessary for evaluating contributions to good health and disease prevention from whole-wheat consumption.     

Phytochemicals and dietary fiber components in rye varieties in the HEALTHGRAIN Diversity Screen

Ten rye varieties grown in one location were analyzed for their contents of dietary fiber (arabinoxylan and beta-glucan) and phytochemicals (folate, tocols, phenolic acids, alkylresorcinols, and sterols). The varieties included old and modern varieties from five European countries. Significant differences were observed in the contents of all phytochemicals in whole grains and in the fiber contents in the flour and bran. The old French varieties Haute Loire and Queyras had high contents of most phytochemicals, whereas the Polish varieties Dankowskie-Zlote and Warko were relatively poor in phytochemicals. The varieties with a high content of folate tended to have low alkylresorcinol contents and vice versa. Furthermore, high contents of arabinoxylans were associated with high contents in tocols and sterols. The 10 selected rye samples comprising old populations and old and modern varieties from different ecological regions of Europe demonstrate high natural variation in their composition and show that landraces and old populations are useful genetic resources for plant breeding. The contents of single phytochemicals can likely be affected by breeding, and they may be adjusted by the right selection of genotype.     

Analysis of Genotype,Environment, and Their Interaction Effects on the Phytochemicals and Antioxidant Capacities of Red Rice (Oryza sativa L.)

Fourteen red rice varieties were planted in two locations during summer (Hangzhou) and winter (Hainan) to study the effect of genotype and environment on the phytochemicals and antioxidant capacities of rice grain. B‐type proanthocyanidins in red rice were detected by LC‐MS/MS and quantified by using the vanillin assay. Analysis of variance showed that total phenolic content (TPC), total flavonoid content (TFC) and 2,2′‐azino‐bis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical scavenging capacity were mainly affected by environmental factors, which accounted for more than 60% of the total variance. However, total proanthocyanidin content (TPAC) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging capacity were equally affected by both genotype and environment. The genotype × environment effects were significant for all traits. The pairwise correlations among TPC, TFC, TPAC, ABTS, and DPPH were also significant (r > 0.900, P < 0.001). Principal component analysis identified the genotypes that had higher contents of antioxidants and more stability across environments. This study showed that indirect selection of a simple trait (i.e., TPC) is an effective way to select rice high in antioxidant capacity in breeding programs. This study also suggests that rice should be produced specifically in a certain environment for the end user to minimize the variation in the functional properties and maximize their contents.     

Genetic resources for arcelin,a stored product insect antimetabolic protein from various accessions of pulses of Leguminosae

Pulses, the most important source of high-quality dietary proteins are prone to serious damage due to insect pests, of which 20–25% damage was inflicted by stored product bruchid pest, Callosobruchus maculatus (F.). An alternative approach to control these insect pests apart from fumigation is the use of primary metabolites involved in plant defense mechanism. In the present study, one of such molecule, an insect anti-metabolic protein, arcelin was screened from nine different wild and cultivated varieties of pulses. Insect feeding bioassay with C. maculatus revealed that seven of these seed varieties were not infested by the bruchid insect pest. Additionally, the presence of arcelin polypeptide subunits (27–42 kDa) was demonstrated in the seeds of Phaseolus limensis L. and Phaseolus lunatus L. whereas, an insignificant amount was observed in Lablab purpureus L. Besides, seed kernel extract of P. lunatus registered a highest HA activity against native human-A erythrocytes and this activity was increased to several fold when the erythrocytes were treated with proteolytic enzymes revealing the characteristic feature of arcelin. Similarly, HA activity was observed when rat and mouse erythrocyte was treated with pronase, whereas, no titer value was observed upon treatment with its native erythrocytes. Of the 24 carbohydrates tested, only acetylated amino sugar, N-acetyl-d-galactosamine inhibited the agglutinating activity of crude seed extract against human-A RBC. Among the three glycoproteins tested, none were found potential to competitively inhibit the agglutination activity against human-A erythrocytes. Overall, this study reveals the presence of arcelin, an anti-metabolic insecticidal protein in seeds of P. lunatus.     

Acid pulses from snowmelt at acidic cone pond,New Hampshire

A study was undertaken to examine whether ‘acid pulses’ from snowmelt created permanent changes in a pond's chemistry. Water samples were collected from clearwater acidic Cone Pond in the White Mountain National Forest, New Hampshire. The pond, inlet, and outlet were intensively sampled throughout winter and early spring 1983–84. Thaws brought more H⁺ into upper waters of the pond, but most was gone within a week. In contrast, SO₄ ^2? and Al showed dilution with increased streamflow into the pond, and NO₃ ^? was only detected in ice, slush, and surface waters. Bottom waters were anoxic throughout the winter and had pH 6.0 compared to 4.7 for most of the water column. Alkalinity at the bottom rose from 0 in November 1983 to 190 μeq L^?1 in April 1984. Between November and April the pond gained Al but lost SO₄ ^2? and H⁺. Most of the Al gain came after ice-out when loading through the inlet increased, but during the final snowmelt a temporary increase in Al concentration was also seen throughout the water column.     

Preclinical evaluation of rapeseed, raspberry, and pine bark phenolics for health related effects

Rapeseed, raspberry, and pine bark are promising bioactive sources of plant phenolics selected from among ca. 100 previously screened plant materials for in vitro preclinical evaluation of health related effects. Phenolic extracts and isolated fractions of the selected materials were investigated for antioxidant, antimicrobial, antiinflammatory, and antimutagenic properties as well as for cell permeability. It was shown that rapeseed and pine bark phenolics and raspberry anthocyanins were good or excellent antioxidants toward oxidation of phosphatidylcholine membrane (liposomes), rapeseed oil (crude) phenolics were effective radical scavengers (DPPH test), and both raspberry and pine bark phenolics inhibited LDL oxidation. Rapeseed oil phenolics, principally vinylsyringol, raspberry anthocyanins, and pinoresinol and matairesinol, the principal components of pine bark phenolic isolate, were effective against formation of the proinflammatory mediator, prostaglandin E(2). Raspberry ellagitannins inhibited the growth of Proteus mirabilis and Klebsiella oxytoca. Pine bark and rapeseed had minor effects on the permeability of model drugs in Caco-2 experiments. None of the tested extracts were mutagenic nor toxic to Caco-2 cells or macrophages. Thus, phenolic isolates from rapeseed, raspberry, and pine bark and are safe and bioactive for possible food applications including functional foods intended for health benefit.     

Carbon pulses but not phosphorus pulses are related to decreases in microbial biomass during repeated drying and rewetting of soils

Drying and rewetting cycles are known to be important for the turnover of carbon (C) in soil, but less is known about the turnover of phosphorus (P) and its relation to C cycling. In this study the effects of repeated drying-rewetting (DRW) cycles on phosphorus (P) and carbon (C) pulses and microbial biomass were investigated. Soil (Chromic Luvisol) was amended with different C substrates (glucose, cellulose, starch; 2.5 g C kg⁻¹) to manipulate the size and community composition of the microbial biomass, thereby altering P mineralisation and immobilisation and the forms and availability of P. Subsequently, soils were either subjected to three DRW cycles (1 week dry/1 week moist) or incubated at constant water content (70% water filled pore space). Rewetting dry soil always produced an immediate pulse in respiration, between 2 and 10 times the basal rates of the moist incubated controls, but respiration pulses decreased with consecutive DRW cycles. DRW increased total CO₂ production in glucose and starch amended and non-amended soils, but decreased it in cellulose amended soil. Large differences between the soils persisted when respiration was expressed per unit of microbial biomass. In all soils, a large reduction in microbial biomass (C and P) occurred after the first DRW event, and microbial C and P remained lower than in the moist control. Pulses in extractable organic C (EOC) after rewetting were related to changes in microbial C only during the first DRW cycle; EOC concentrations were similar in all soils despite large differences in microbial C and respiration rates. Up to 7 mg kg⁻¹ of resin extractable P (P_resin) was released after rewetting, representing a 35-40% increase in P availability. However, the pulse in P_resin had disappeared after 7 d of moist incubation. Unlike respiration and reductions in microbial P due to DRW, pulses in P_resin increased during subsequent DRW cycles, indicating that the source of the P pulse was probably not the microbial biomass. Microbial community composition as indicated by fatty acid methyl ester (FAME) analysis showed that in amended soils, DRW resulted in a reduction in fungi and an increase in Gram-positive bacteria. In contrast, the microbial community in the non-amended soil was not altered by DRW. The non-selective reduction in the microbial community in the non-amended soil suggests that indigenous microbial communities may be more resilient to DRW. In conclusion, DRW cycles result in C and P pulses and alter the microbial community composition. Carbon pulses but not phosphorus pulses are related to changes in microbial biomass. The transient pulses in available P could be important for P availability in soils under Mediterranean climates.     

The role of mentoring in public health nutrition workforce development. Perspectives of advanced-level practitioners

OBJECTIVE: To explore the nature, role and utility of mentoring in the development of competence in advanced-level Australian public health nutritionists. DESIGN: Qualitative study using in-depth interviews. SUBJECTS AND SETTING: Eighteen advanced-level public health nutritionists working in academic and practice settings in Australia. RESULTS: The attributes and career pathways of the subjects were consistent with previous findings. Dissatisfaction with clinical practice was a key reason for choosing a career in public health. Experiential learning, postgraduate education and mentoring from both peers and senior colleagues were the most significant contributors to competency development. The subjects supported mentoring as an important strategy for public health nutrition workforce development and articulated the characteristics and models important for mentoring relationships in public health nutrition. CONCLUSIONS: The present study suggests mentoring was an important part of competency development for advanced-level public health and community nutritionists in Australia. Mentoring programmes based on experiential learning may assist in developing public health nutrition workforce competence.     

Organochlorine pesticide residues in different Indian cereals, pulses, spices, vegetables, fruits, milk, butter, Deshi ghee, and edible oils.

A total of 244 samples of cereals (wheat flour, rice, and maize), pulses (arhar, moong, gram, lentil, and black gram), spices (turmeric, chili, coriander, and black pepper), vegetables (potato, onion, spinach, cabbage, brinjal, and tomato), fruits (mango, guava, apple, and grape), milk, butter, Deshi ghee, and edible oils (vegetable, mustard, groundnut, and sesame) collected from different cities of Northern Province (Utter Pradesh) were analyzed by gas liquid chromatography for the presence of organochlorine pesticide residues. Residues of hexachlorocyclohexane (HCH) and 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane (DDT) were detected in about 85% of the total samples of cereals, spices, milk, butter, Deshi ghee, and edible oils analyzed in the present study. However, the residue levels were either very small (less than 0.06 ppm) or not detected at all in pulses, vegetables, and fruits as compared with very high concentrations in wheat flour (4.42 and 0.12 ppm), butter (1.19 and 4.85 ppm), mustard oil (1.26 and 2.42 ppm), Deshi ghee (1.10 and 3.84 ppm), vegetable oil (1.02 and 0.59 ppm), groundnut oil (0.51 and 1.49 ppm), and chili (0.48 and 1.92 ppm). The levels of HCH and DDT residues detected in rice, maize, turmeric, corlander, black pepper, and all the vegetables and fruits were also lower than those found in wheat flour, oil, and fat samples analyzed in the present study. These findings suggest that a restricted and controlled use of such persistent pesticides may be useful for decreasing their contamination levels in different food items.     

A role for pectin in the control of cell expansion

Uptake of nutrients and water depends on the growth of roots through elongation of individual cells near the root tip. Many of the numerous components of Type I primary cell walls, those of dicotyledons and monocotyledons other than grasses (Poaceae), have been determined, and many hypotheses have been proposed for the control of cell expansion. This important aspect of plant growth still needs elucidation, however. A model is proposed in which pectin, which occurs as a calcium (Ca) pectate gel between the load-bearing cellulose microfibrils and xyloglucan (XG) chains, controls the rate at which cells expand. It is considered that the increasing tension generated by the expanding cell is transmitted to interlocked XG chains and cellulose microfibrils. The resulting deformation of the embedded Ca pectate gel elicits the excretion of protons from the cytoplasm, possibly via compounds such as cell wall-associated kinases, that weakens the Ca pectate gel, permitting slippage of XG molecules through the action of expansin. Further slippage is prevented by deformation of the pectic gel, proton diffusion, and the transfer of residual tension to adjacent XG chains. Evidence for this model is based on the effects of pH, Ca, and aluminum (Al) on root elongation and on the reactions of these cations with Ca pectate. This model allows for genetic selection of plants and adaptation of individual plants to root environmental conditions.     

Plant growth-promoting bacteria in the rhizo- and endosphere of plants: Their role, colonization, mechanisms involved and prospects for utilization

In both managed and natural ecosystems, beneficial plant-associated bacteria play a key role in supporting and/or increasing plant health and growth. Plant growth-promoting bacteria (PGPB) can be applied in agricultural production or for the phytoremediation of pollutants. However, because of their capacity to confer plant beneficial effects, efficient colonization of the plant environment is of utmost importance. The majority of plant-associated bacteria derives from the soil environment. They may migrate to the rhizosphere and subsequently the rhizoplane of their hosts before they are able to show beneficial effects. Some rhizoplane colonizing bacteria can also penetrate plant roots, and some strains may move to aerial plant parts, with a decreasing bacterial density in comparison to rhizosphere or root colonizing populations. A better understanding on colonization processes has been obtained mostly by microscopic visualisation as well as by analysing the characteristics of mutants carrying disfunctional genes potentially involved in colonization. In this review we describe the individual steps of plant colonization and survey the known mechanisms responsible for rhizosphere and endophytic competence. The understanding of colonization processes is important to better predict how bacteria interact with plants and whether they are likely to establish themselves in the plant environment after field application as biofertilisers or biocontrol agents.     

Rewetting CO2 pulses in Australian agricultural soils and the influence of soil properties

An incubation study determined the effect of one dry–rewetting (DRW) event on the turnover of carbon (C), phosphorus (P) and nitrogen (N). Thirty-two soils were collected from different climatic regions of southern Australia, varying in soil type, land use and agronomic management history. We hypothesised that respiration and nutrient pulses are related to soil physio-chemical properties. Respiration (CO₂ release) was measured intensively for 90 h after rewetting. C mineralisation (C _min) model fitting was used to describe the amount of mineralisable C (Co_90 h) and the proportional mineralisation rate (k). Compared to constantly moist soils, 13 soils showed increases in both Co_90 h and k, indicating that DRW increased the amount of mineralisable C and the rate at which C was mineralised over the 90-h period. In 17 soils, k was increased but not Co_90 h, showing an increase in C mineralisation rate but no change in the amount of mineralisable C. Two soils showed a reduction in k with no change in Co_90 h, possibly due to low C contents and small microbial biomass. Only one soil exhibited no change in either Co_90 h or k. Multiple linear regression analysis indicated that the magnitude of the increase in mineralisable C in response to the DRW event (∆Co_90 h = Co_90 h DRW − Co_90 h moist) was primarily explained by clay content (39%); however, inclusion of nine soil physio-chemical properties explained more of the variation in ∆Co_90 h than any of the properties alone. Five of the nine physio-chemical variables present in the multiple-regression model were related to C content or composition. Pulses in available N and P were not related to ∆Co_90 h.