Cadmium uptake by onions, lettuce and spinach in New Zealand: Implications for management to meet regulatory limits.

Paired soil and plant samples collected from the main commercial growing areas for onions (Allium cepa), lettuce (Lactuca sativa) and spinach (Spinacia olearacea) in New Zealand were used to assess the influence of plant and soil factors on cadmium (Cd) uptake in these crops. Differences in Cd concentration between eight lettuce sub-types were not consistent across sites, nor were differences in Cd concentrations in three crisphead cultivars assessed at two sites. Similarly, differences in Cd concentrations between four onion cultivars were inconsistent across sites. Mean lettuce Cd concentrations in eight lettuce varieties (range 0.005-0.034 mg∙kg-1 (fresh weight, FW) were markedly lower than those in baby leaf and bunching spinach, (range 0.005-0.19 mg∙kg-1 FW). Significant regional variation was observed in Cd concentrations in one onion cultivar (mean range 0.007-0.05 mg∙kg-1 FW). Soil Cd concentration, pH and region were statistically significant predictors of onion Cd concentration, explaining low (38% for soil Cd and pH) to moderate (50% for all three parameters) percentage of the variation. Soil Cd concentration and exchangeable magnesium or total carbon were statistically significant predictors of Cd concentration in baby leaf and bunching spinach, respectively, explaining a moderate percentage (49% and 42%) of the variation in Cd concentration. Increasing pH and soil carbon may assist in minimising Cd uptake in onion and bunching spinach, respectively. The low to moderate proportion of explained variation is partly attributable to the narrow range in some measured soil properties and indicates factors other than those assessed are influencing plant uptake. This highlights a challenge in using these relationships to develop risk-based soil guideline values to support compliance with food standards. Similarly, the inconsistency in Cd concentrations in different cultivars across sites highlights the need for multi-site assessments to confirm the low Cd accumulation status of different cultivars.

Biowastes to augment the essential oil production of Leptospermum scoparium and Kunzea robusta in low-fertility soil.

Biowastes are unwanted materials of biological origin. They include biosolids, dairy shed effluent, and sawdust. When applied to soil, biowastes can provide plant nutrients, but also introduce heavy metals, pathogens, or xenobiotics. Biowastes could improve degraded or low-fertility soils and generate revenue through the production of non-food products such as essential oils. We grew New Zealand native plants, manuka (Leptospermum scoparium J.R. Forst & G. Forst) and kanuka (Kunzea robusta de Lange & Toelken) in series of greenhouse experiments in low-to-medium-fertility soils (Bideford clay loam, Lismore stony silt loam, and Pawson silt loam) amended with either biosolids (up to 13500 kg N ha-1 equiv.), biosolids + sawdust (1:0.5-1250 kg N ha-1 equiv.) and dairy shed effluent (200 kg N ha-1 equiv.). Two types of biosolids from Kaikoura (KB) and Christchurch City Council (CB) were used in the experiments. CB (1500 kg N ha-1 equiv.) and dairy shed effluent (200 kg N ha-1 equiv.) increased the biomass of L. scoparium by up to 120% and 31%, and K. robusta by up to 170% and 34%, respectively. Adding sawdust to KB increased the biomass of L. scoparium and K. robusta although it offset the L. scoparium growth increase in the KB-only treatment. The growth response of K. robusta to biowastes was greater than L. scoparium with oil production in K. robusta increasing by up to 211% when 1500 kg N ha-1 equiv. of CB was applied to Lismore stony silt loam. Generally, the treatments had a negligible effect on oil concentration in all the soil types, except for the KB + sawdust treatment, which increased the oil concentration by 82%. Most of the EOs' major components were unaffected by biowaste addition in the soils, although some components increased in the Bideford clay loam following KB and KB + sawdust application. Biosolids increased foliar concentrations of Zn, Cu, and Cd, but these were below risk-threshold concentrations. Applying CB (up to 1500 kg N ha-1 equiv.) to low-fertility soils is recommended to establish ecosystems dominated by L. scoparium and K. robusta that annually would produce ca. 100 kg ha-1 of EOs worth US$ 26k and 24k, respectively. Adding sawdust to CB could have environmental benefits through reduction of N leaching. Field trials are warranted to elucidate critical ecological variables and production economics in biowaste management.

The potential of L. scoparium, K. robusta and P. radiata to mitigate N-losses in silvopastural systems.

Silvopastoral systems aim to enhance economic, cultural and social principles by sustainably combining forest management with agriculture. In these typically high-nitrogen (N) environments, plant species selection can profoundly influence N fluxes. For grazed pastures, plants may be exposed to urine patches that have received the equivalent of up to 1000 kg N ha-1. We aimed to determine the growth and N fluxes in three potential trees that may be used in silvopastoral systems: L. scoparium, K. robusta and P. radiata. Plants were grown in a greenhouse lysimeter experiment, with controlled irrigation and temperature and exposed to N at rates of 200 kg ha-1 equiv. for 15 weeks, followed by the addition of 800 kg ha-1 N equiv, to simulate a urine patch. Urea produced a positive growth response of all plant species. Treatments containing L. scoparium and K. robusta leached lower amounts of nitrate (NO3-) (2 kg ha-1 NO3-) compared to P. radiata (53 kg ha-1). Measurements of N2O over 20 days after the application of 800 kg N ha-1 indicated an inhibitory effect of L. scoparium and K. robusta on denitrification, hence loss of N via N2O. Both L. scoparium and K. robusta demonstrated that they have potential to reduce N-losses in silvopastural systems, while producing valuable biomass.

Response of Leptospermum scoparium, Kunzea robusta and Pinus radiata to contrasting biowastes.

The myrtaceae family has a cosmopolitan distribution and includes the Australasian native species Leptospermum scoparium (manuka) and Kunzea robusta (kanuka), which are of economic interest for the production of high-value honey and essential oils. Potentially, these species could be established on low-fertility or degraded soils that have been amended with biowastes, including biosolids and sawdust. We aimed to determine the effect of these biowastes on nitrate leaching and the growth and chemical composition of these plant species compared to Pinus radiata (pine), a common forestry species. The addition of biosolids (1250kgNha-1 equiv.) increased the total dry biomass of manuka, kanuka, and pine by 117, 90, and 86% respectively. Mixing sawdust with biosolids stimulated growth of manuka (52%), kanuka (121%) but not pine. Biosolids increased plant uptake of N, P, and trace elements, but not to levels of concern. Nitrate leaching from all treatments was negligible (<2kgha-1).

Production of Biomass Crops Using Biowastes on Low-Fertility Soil: 1. Influence of Biowastes on Plant and Soil Quality.

Land application of biosolids to low-fertility soil can improve soil quality by increasing concentrations of macronutrients and trace elements. Mixing biosolids with sawdust could reduce the risks of contaminant accumulation posed by rebuilding soils using biosolids alone. We aimed to determine the effects of biosolids and biosolids-sawdust on the plant quality and chemical composition of sorghum, rapeseed, and ryegrass. Plants were grown in a greenhouse over a 5-mo period in a low-fertility soil amended with biosolids (1250 kg N ha), biosolids-sawdust (0.5:1), or urea (200 kg N ha). Biosolids application increased the biomass of sorghum, rapeseed, and ryegrass up to 14.0, 11.9, and 4.1 t ha eq, respectively. Mixing sawdust with biosolids resulted in a growth response similar to biosolids treatments in rapeseed but nullified the effect of biosolids in sorghum. Urea fertilization provided insufficient nutrients to promote rapeseed growth and seed production, whereas seed yields after biosolids application were 2.5 t ha. Biosolids and biosolids-sawdust application enhanced plant quality by increasing element concentrations, especially Zn, and potentially toxic elements (Cd, Cr, Ni) did not exceed food safety standards. An application of 50 t ha of biosolids, equivalent to 1250 kg N ha, did not exceed current soil limits of Cu, Zn, and Cd and hence was effective in rebuilding soil without accumulating contaminants. The effect of mixing sawdust with biosolids varies with plant species but can further enhance plant nutrient quality in biomass and seeds, especially P, Cu, Zn, Mn, Fe, S, and Na.

Production of Biomass Crops Using Biowastes on Low-Fertility Soil: 2. Effect of Biowastes on Nitrogen Transformation Processes.

Increasing production of biowastes, particularly biosolids (sewage sludge), requires sustainable management strategies for their disposal. Biosolids can contain high concentrations of nutrients; hence, land application can have positive effects on plant growth and soil fertility, especially when applied to degraded soils. However, high rates of biosolids application may result in excessive nitrogen (N) leaching, which can be mitigated by blending biosolids with other biowastes, such as sawdust. We aimed to determine the effects of biosolids and sawdust on growth and N uptake by sorghum, rapeseed, and ryegrass as well as N losses via leaching. Plants were grown in a greenhouse over a 5-mo period in a low-fertility soil amended with biosolids (1250 kg N ha), biosolids-sawdust (0.5:1), or urea (200 kg N ha). Urea application increased biomass production of sorghum and ryegrass but proved insufficient for rapeseed on low-fertility soil. Biosolids application increased plant N concentrations in ryegrass and rapeseed and increased N uptake into the seeds of sorghum, increasing seed quality. Biosolids application did result in lower N leaching compared with urea, irrespective of plant species, and N leaching was unaffected by mixing the biosolids with sawdust. There was an indication of biological nitrification inhibition in the rhizosphere of sorghum. Rapeseed had similar growth and N uptake into biomass in biosolids and biosolids-sawdust treatments and hence was the most promising species with regard to recycling fresh sawdust in combination with high rates of biosolids on low-fertility soil.

Risk assessment of vegetables irrigated with arsenic-contaminated water.

Arsenic (As) contaminated water is used in South Asian countries to irrigate food crops, but the subsequent uptake of As by vegetables and associated human health risk is poorly understood. We used a pot trial to determine the As uptake of four vegetable species (carrot, radish, spinach and tomato) with As irrigation levels ranging from 50 to 1000 µg L(-1) and two irrigation techniques, non-flooded (70% field capacity for all studied vegetables), and flooded (110% field capacity initially followed by aerobic till next irrigation) for carrot and spinach only. Only the 1000 µg As L(-1) treatment showed a significant increase of As concentration in the vegetables over all other treatments (P < 0.05). The distribution of As in vegetable tissues was species dependent; As was mainly found in the roots of tomato and spinach, but accumulated in the leaves and skin of root crops. There was a higher concentration of As in the vegetables grown under flood irrigation relative to non-flood irrigation. The trend of As bioaccumulation was spinach > tomato > radish > carrot. The As concentration in spinach leaves exceeded the Chinese maximum permissible concentration for inorganic As (0.05 µg g(-1) fresh weight) by a factor of 1.6 to 6.4 times. No other vegetables recorded an As concentration that exceeded this threshold. The USEPA parameters hazard quotient and cancer risk were calculated for adults and adolescents. A hazard quotient value greater than 1 and a cancer risk value above the highest target value of 10(-4) confirms potential risk to humans from ingestion of spinach leaves. In our study, spinach presents a direct risk to human health where flood irrigated with water containing an arsenic concentration greater than 50 µg As L(-1).

Biochar for the mitigation of nitrate leaching from soil amended with biosolids.

Countries with sewage treatment plants produce on average 27 kg of dried biosolids/person/yr. Concerns about nitrate leaching limit the rate at which biosolids are added to soil. We sought to determine whether biochar, a form of charcoal that is added to soil, could reduce nitrate leaching from biosolids amended soil. We set up 24 (0.5 m × 0.75 m) lysimeters, filled with two soil types (Templeton Silt Loam and Ashley Dene silt loam) and amended with combinations of biochar (102 t/ha equivalent) and biosolids (600 and 1200 kg N/ha equivalent). Pasture and leachates were sampled over 5 months. Nitrate leaching from biochar plus biosolids amended soils were reduced to levels at or below the control treatments. Pasture N concentrations were similarly affected by biochar addition. Future research should focus on unravelling the mechanism responsible for the change in the nitrogen cycle in soils amended with biosolids and biochar.

Pyruvate dehydrogenase phosphatase 1 (PDP1) null mutation produces a lethal infantile phenotype.

Pyruvate dehydrogenase phosphatase deficiency has previously only been confirmed at the molecular level in two brothers and two breeds of dog with exercise intolerance. A female patient, who died at 6 months, presented with lactic acidemia in the neonatal period with serum lactate levels ranging from 2.5 to 17 mM. Failure of dichloroacetate to activate the PDH complex in skin fibroblasts was evident, but not in early passages. A homozygous c.277G > T (p.E93X) nonsense mutation in the PDP1 gene was identified in genomic DNA and immunoblotting showed a complete absence of PDP1 protein in mitochondria. Native PDHC activity could be restored by the addition of either recombinant PDP1 or PDP2. This highlights the role of PDP2, the second phosphatase isoform, in PDP1-deficient patients for the first time. We conclude that the severity of the clinical course associated with PDP1 deficiency can be quite variable depending on the exact nature of the molecular defect.

Pyruvate dehydrogenase deficiency presenting as intermittent isolated acute ataxia.

OBJECTIVE: The aim of this study is to report and emphasize unusual presentations of pyruvate dehydrogenase (PDH) deficiency (OMIM 312170). METHODS: PDH activity and PDHA1 gene were studied in two siblings presenting with intermittent ataxia in childhood. Similar presentations in reported PDH-deficient patients were searched for using the Medline database. RESULTS: Both patients had PDH deficiency caused by a new mutation (G585C) in the PDHA1 gene, which is predicted to replace a highly conserved glycine at codon 195 by alanine. Although this mutation lies within the thiamine pyrophosphate binding domain, there was no thiamine responsiveness IN VIVO. The patients presented recurrent episodes of acute isolated ataxia in infancy. Both had normal blood and CSF lactate levels. Although symptoms initially resolved between episodes during the first decade, both patients subsequently worsened and developed progressive and severe encephalopathy, leading to death in their twenties. The spectrum of intermittent presentations in PDH deficiency includes episodic ataxia, intermittent peripheral weakness, recurrent dystonia and extrapyramidal movement disorders. CONCLUSIONS: PDH deficiency should be considered in patients with unexplained intermittent and recurrent acute neurological symptoms. Long-term prognosis and outcome remain uncertain. PDH deficiency can occur even with normal CSF lactate concentration.

A novel mitochondrial DNA mutation in COX1 leads to strokes, seizures, and lactic acidosis.

Cytochrome c oxidase (COX) is the terminal enzyme of the respiratory chain, with subunits originating both from the mitochondrial and nuclear genome. An eleven-year-old female presented initially with a seizure followed two months later with tonic-clonic seizures, weakness and aphasia. MRI of the cerebral hemispheres showed multiple infarcts. Previous history suggested gross and fine motor control deficits with learning difficulties. A muscle biopsy showed a specific decrease of COX staining in all fibres and pleomorphic mitochondria. Respiratory chain studies confirmed an isolated complex IV defect in muscle, whilst fibroblasts showed an initial COX activity below normal which rapidly came up to the normal range on culture. Sequencing of mtDNA revealed an heteroplasmic m.7023G>A mutation in the COX1 gene, with levels of 96% in muscle, 70% in blood and 50% in the initial skin fibroblast culture dropping to 10% in later passages. The mutation was present in a critical region of the COX1 gene, the V374M change being close to the two histidine residues His376 and His378 co-ordinating with the heme a and a (3), and His367 which co-ordinates a magnesium ion. This case highlights that a MELAS-like syndrome can occur with isolated COX deficiency.

Poplar for the phytomanagement of boron contaminated sites.

Boron (B) is a widespread environmental contaminant that is mobile relative to other trace elements. We investigated the potential of hybrid poplar (Populus sp.) for B phytomanagement using a lysimeter experiment and a field trial on B-contaminated wood-waste. In both studies, poplars enhanced evapotranspiration from the wood-waste, reduced B leaching, and accumulated B in the aerial portions of the tree. When grown in a substrate containing 30 mg/kg B, poplar leaves had an average B concentration of 845 mg/kg, while the stems contained 21 mg/kg B. Leaf B concentrations increased linearly with leaf age. A decomposition experiment revealed that abscised leaves released 14% of their B during the winter months. Fertiliser application enhanced tree growth without decreasing the leaf B concentrations. Harvesting alternate rows of trees on a contaminated site would reduce leaching from the site while removing B. Harvested plant material may provide bioenergy, stock fodder, or an amendment for B-deficient soils.

Comparison of static first hyperpolarizabilities calculated with various quantum mechanical methods.

The prediction of nonlinear electro-optic (EO) behavior of molecules with quantum methods is the first step in the development of organic-based electro-optic devices. Typical EO molecules may require calculations with several hundred electrons, which prevents all but the fastest methods (semiempirical and density functional theory (DFT)) from being used for EO estimation. To test the reliability of these methods, we compare dipole moments, polarizabilities, and first-order hyperpolarizabilities for a wide range of structures of experimental interest with Hartree-Fock (HF), intermediate neglect of differential overlap (INDO), and DFT methods. The relative merits of molecules are consistently predictable with every method.

Influence of isomerization on nonlinear optical properties of molecules.

The influence of rotational and geometrical isomerism on the nonlinear optical (NLO) properties, specifically the first-order hyperpolarizability beta, of chromophores of current interest has been investigated with density functional theory (DFT). In the first of this two-part study, the rotational isomerism of a linear chromophore was explored. Calculation of the torsion potentials about two of the rotatable and conformation-changing single bonds in a chromophore demonstrated the near equality of the molecular energies at 0 degrees and 180 degrees rotational angles. To explore the consequences of this near conformational energy degeneracy to NLO behavior, the eight low energy rotational isomers of FTC [Robinson, B. H.; et al. Chem. Phys. 1999, 245, 35] were investigated. This study provides the first-reported DFT-based calculation of the statistical mechanical average of beta over the conformational space of a molecule having substantial nonlinear optical behavior. The influence of the solvent reaction field on rotameric populations and on the beta tensor is reported. In the second part, two molecules having two donors and two acceptors bonded respectively in ortho and meta positions on a central benzene ring are shown to have substantially different beta tensors. These two so-called molecular Xs have different highest occupied molecular orbital to lowest unoccupied molecular orbital (HOMO-LUMO) distributions, and consistent with expectations, it is found that the larger beta(zzz) is associated with a large spatial asymmetry between the HOMOs and LUMOs. Large hyperpolarizability correlates with the HOMO concentrated on the donor groups and the LUMO on the acceptor groups.

Pyruvate dehydrogenase phosphatase deficiency: orphan disease or an under-diagnosed condition?

Pyruvate dehydrogenase phosphatase (PDP) is an enzyme which regulates the activity of the pyruvate dehydrogenase complex (PDHc). In the past, PDHc deficiency has been attributed to mutations in the complex itself and the regulatory enzymes have not been considered. We have recently reported the first mutation in PDP1, one of the two isoforms of PDP, which results in severe exercise intolerance and mild developmental delay in patients. This novel process of aberrant pyruvate metabolism opens up a new avenue for investigation into PDHc deficiency, that has hitherto been underappreciated.

Computational identification of human mitochondrial proteins based on homology to yeast mitochondrially targeted proteins.

MOTIVATION: Patients with defects of the mitochondrial respiratory chain due to mutations in nuclear genes are often undiagnosable due to the lack of information about the role of these genes. We therefore sought to produce a novel dataset of human nuclear-encoded mitochondrial proteins. RESULTS: We have used the web-based computer program Mitoprot to predict which proteins in the Saccharomyces cerevisiae genome are targeted to mitochondria. We then used this protein dataset to identify the homologous human proteins in the Unigene database using TBLASTN from NCBI. Human proteins with an Expectation value <10(-5) and an Identity >30% were accepted as true homologues of the yeast proteins. These human proteins were then reanalyzed with Mitoprot. The final set of proteins comprises a dataset of 361 human mitochondrially targeted proteins with homology to all S.cerevisiae mitochondrially targeted proteins. One hundred twenty eight of these proteins are novel and are of unknown function. SUPPLEMENTARY INFORMATION: Supplementary tables will be available from http://www.sickkids.ca/Robinsonlab/

Novel SCO2 mutation (G1521A) presenting as a spinal muscular atrophy type I phenotype.

Rare cases of suspected spinal muscular atrophy (SMA) have been found to have cytochrome c oxidase (COX) deficiency. To date, four cases with SMA features have been reported in children with mutations in the synthesis of cytochrome oxidase 2 (SCO2) gene. We report a male neonate who was born hypotonic, with persistent lactic acidosis, spontaneous activity with EMG testing, development of respiratory distress in the first few hours of life, and died at 30 days of age with progressive cardiomyopathy. Testing for survival motor neurone (smn) and NAIP deletions were negative and a skeletal muscle biopsy showed neurogenic features with severe reductions of COX enzymatic and histochemical staining intensity. Post-mortem muscle, heart, and liver biopsies showed severe, moderate, and mild reductions in COX activity, respectively, with parallel findings in the protein content for the mitochondrial DNA (COII) and nuclear DNA (COIV) encoded subunits. DNA sequencing of exon 2 of the SCO2 gene revealed compound heterozygosity with mutations at G1541A (common mutation, E140K) and also at a novel site in the copper binding region (G1521A in the current case (converting a highly conserved cysteine to tyrosine [corrected] (C133Y) [corrected]); mother heterozygous for G1521A; and father heterozygous for G1541A). This case provides strong support that SCO2 mutations can result in neonatal hypotonia with an SMA 1 phenotype. SCO2 mutations should be screened in suspected SMA cases with normal smn mutation analysis and any one of; cardiomyopathy, lactic acidosis, or COX deficiency in muscle.

Nickel and cobalt phytoextraction by the hyperaccumulator Berkheya coddii: implications for polymetallic phytomining and phytoremediation.

We investigated the potential of the South African high-biomass Ni hyperaccumulator Berkheya coddii to phytoextract Co and/or Ni from artificial metalliferous media. Plant accumulation of both metals from single-element substrates indicate that the plant/media metal concentration quotient (bioaccumulation coefficient) increases as total metal concentrations increase. Cobalt was readily taken up by B. coddii with and without the presence of Ni. Nickel uptake was, however, inhibited by the presence of an equal concentration of Co. Bioaccumulation coefficients of Ni and Co for the single element substrates (total metal concentration of 1000 micrograms g-1) were 100 and 50, respectively. Cobalt phytotoxicity was observed above a total Co concentration in plant growth media of 20 micrograms g-1. Elevated Co concentrations significantly decreased the biomass production of B. coddii without affecting the bioaccumulation coefficients. The mixed Ni-Co substrate produced bioaccumulation coefficients of 22 for both Ni and Co. Cobalt phytotoxicity in mixed Ni-Co substrate occurred above a total Co concentration of 15 micrograms g-1. When grown in the presence of both Ni and Co, the bioaccumulation coefficients of each metal were reduced, as compared to single-element substrate. This may indicate competition for binding sites in the root zone. The interference relationship between Ni and Co uptake demonstrated by B. coddii suggests a significant limitation to phytoextraction where both metals are present.

Sequence-dependent dynamics of duplex DNA: the applicability of a dinucleotide model.

The short-time (submicrosecond) bending dynamics of duplex DNA were measured to determine the effect of sequence on dynamics. All measurements were obtained from a single site on duplex DNA, using a single, site-specific modified base containing a rigidly tethered, electron paramagnetic resonance active spin probe. The observed dynamics are interpreted in terms of single-step sequence-dependent bending force constants, determined from the mean squared amplitude of bending relative to the end-to-end vector using the modified weakly bending rod model. The bending dynamics at a single site are a function of the sequence of the nucleotides constituting the duplex DNA. We developed and examined several dinucleotide-based models for flexibility. The models indicate that the dominant feature of the dynamics is best explained in terms of purine- and pyrimidine-type steps, although distinction is made among all 10 unique steps: It was found that purine-purine steps (which are the same as pyrimidine-pyrimidine steps) were near average in flexibility, but the pyrimidine-purine steps (5' to 3') were nearly twice as flexible, whereas purine-pyrimidine steps were more than half as flexible as average DNA. Therefore, the range of stepwise flexibility is approximately fourfold and is characterized by both the type of base pair step (pyrimidine/purine combination) and the identity of the bases within the pair (G, A, T, or C). All of the four models considered here underscore the complexity of the dependence of dynamics on DNA sequence with certain sequences not satisfactorily explainable in terms of any dinucleotide model. These findings provide a quantitative basis for interpreting the dynamics and kinetics of DNA-sequence-dependent biological processes, including protein recognition and chromatin packaging.

Mitochondria, oxygen free radicals, and apoptosis.

Reactive oxygen species (ROS) generated by mitochondria are produced as by-products of normal oxidative metabolism. The fate of these species is governed by a number of factors that vary from tissue to tissue in mammals and may be involved in the pathogenesis of disease. Reactive oxygen species are also invoked as agents that are important in the processes which become active in cells undergoing apoptosis. Integration of knowledge surrounding these different aspects of ROS generation is difficult and reveals considerable gaps in our understanding.