Simple and sensitive method for pyrroloquinoline quinone (PQQ) analysis in various foods using liquid chromatography/electrospray-ionization tandem mass spectrometry

Pyrroloquinoline quinone (PQQ) is believed to be an important factor for mammalian growth and development and has, therefore, been declared a vitamin by some researchers. However, this issue remains controversial, and from a nutritional viewpoint, accurate determination of PQQ levels in a variety of foods is very important. Here, we describe a simple, highly sensitive, and highly selective method for quantitative analysis of PQQ. Liquid foods or aqueous extracts of solid foods were analyzed using high-performance liquid chromatography (HPLC) combined with electrospray-ionization (ESI) tandem mass spectrometry (MS/MS). (15)N-labeled PQQ was added to the samples as an internal standard. Quantitative analyses of PQQ were performed by multiple reaction monitoring (MRM) with LC/MS/MS. Free PQQ was detected in almost all food samples in the range 0.19-7.02 ng per g fresh weight (for solid foods) or per mL (liquid foods). This method will enable the rapid and simple determination of PQQ levels in many samples.     

Interaction between manganese and zinc in growth of rice plants

Many instances of antagonistic relationships among various micro-elements and between micro- and macro-elements in their absorption, translocation, and functions in plant growth have been reported (1). In a series of our researches on the effect of micro-elements on the growth of rice plants, it was observed that manganese toxicity symptom of rice plants was markedly reduced by increasing the concentration of zinc in the nutrient sdution. Although it has been reported that manganese toxicity symptoms of crops were reduced by addition of Fe (2,3), A1 (4), Ca (5) and silicate (6) to the medium, there have been few reports of the effect of Zn on Mn toxicity of rice plants. In this paper, the interaction between Mn and Zn in the growth of rice plants was reported.     

Effect of CO2 enrichment on biomass production,photosynthesis, and sink activity in Soybean cv. Bragg and its supernodulating mutant nts 1007

Soybean (Glycine max L. Merr.) cv. Bragg and its supernodulating mutant nts 1007 were grown in pots containing vermiculite with a N-free nutrient solution in order to examine the effect of elevated CO₂ concentration (100+20 Pa CO₂ ) on biomass production, photosynthesis, and biological nitrogen fixation. The whole plant weight increase in Bragg was higher than in the mutant at a high CO₂ concentration. Apparent photosynthetic activities of the upper leaves in both Bragg and the mutant increased up to 14 d after treatment initiation by the CO₂ enrichment and thereafter decreased to some extent. Both leaf area and leaf thickness of Bragg increased more than in nts 1007. With the elevated CO₂ concentration, biological nitrogen fixation (BNF) also responded in the same manner as biomass production in both Bragg and nts 1007. The increase of BNF in Bragg was largely due to an increase in nodule weight. Starch contents in the leaves of both Bragg and the mutant increased significantly by CO₂ enrichment, with a higher increase in Bragg than in its mutant. Sugar content in leaf differed only slightly in both Bragg and the mutant. N content in leaf decreased in both Bragg and its mutant, with the decrease being more pronounced in Bragg. However, in other plant parts (roots, stem, and petiole + pods), N content increased in the mutant while in Bragg, it decreased in the pod. N accumulation rate was higher in Bragg than in the mutant and increased more in Bragg than in the mutant by CO₂ enrichment. The ureide content in leaf decreased in Bragg but increased in the mutant by elevated CO₂ concentration. In the nodules, ureide content increased in both Bragg and the mutant by CO₂ enrichment. Based on these results, it is suggested that in terms of biomass production and photosynthetic rate, Bragg responded more to elevated CO₂ concentration than its mutant nts 1007. The alleviation of the stunted vegetative growth of the mutant by CO₂ enrichment was limited despite the significant increase in the photosynthetic activity, presumably due to the limitation of sink activity in the growing parts and not to insufficient supply of N through BNF.     

Development of immune assay system for both CpG and non-CpG DNA from lactic acid bacteria using a transfectant of swine Toll-like receptor 9

Bacterial DNA is expected to be a potent immune stimulating agent to Toll‐like receptor (TLR) 9 expressed cells such as macrophages, monocytes, B lymphocytes, NK cells and dendritic cells. In the present study, we constructed a transfectant of swine TLR9 with mammalian cells. We demonstrated that the transfectant, recognizing both CpG and non‐CpG oligonucleotides from lactic acid bacteria, induced NF‐κB activation by gene reporter assay. These findings indicate that the swine TLR9 transfectant will be highly useful for the screening of immunostimulatory DNA from lactic acid bacteria.     

Chemical characterization of milk oligosaccharides of an African lion (Panthera leo) and a clouded leopard (Neofelis nebulosa)

The Carnivora include the superfamilies Canoidea and Feloidea. In species of Canoidea other than the domestic dog, Canis lupus, the milk contains only traces of lactose and much larger concentrations of oligosaccharides. In this study, lactose was found to be the dominant saccharide in the milk or colostrum of two species of Feloidea, namely the African lion (Panthera leo) and the clouded leopard (Neofelis nebulosa). In addition to lactose, the following oligosaccharides were characterized in the milk of a lion; Neu5Gc(α2‐3)Gal(β1‐4)Glc (3′‐NGc‐SL), Fuc(α1‐2)Gal(β1‐4)Glc (2′‐fucosyllactose) and GalNAc(α1‐3)[Fuc(α1‐2)]Gal(β1‐4)Glc (A‐tetrasaccharide). The colostrum of a clouded leopard contained 3′‐NGc‐SL, Gal(α1‐3)Gal(β1‐4)Glc (isoglobotriose) and A‐tetrasaccharide. These oligosaccharides differ in some respects from those previously identified in another species of Feloidea, the spotted hyena (Crocuta crocuta). These milks contained 3′‐NGc‐SL and A‐tetrasaccharide, while spotted hyena colostrum did not; however, it contained Neu5Ac(α2‐3)Gal(β1‐4)Glc (3′‐NAc‐SL) and Gal(α1‐3)[Fuc(α1‐2)]Gal(β1‐4)Glc (B‐tetrasaccharide).     

Electroretinography using contact lens electrode with built-in light source in dogs

Electroretinography (ERG) is an effective method for the diagnosis of retinal disease. In the dog, dependable ERG recording is difficult without the use of an expensive device like a Ganzfeld full-field stimulator. The International Society for Clinical Electrophysiology of Vision has defined the standard flash stimulus condition (SF) and evaluation of the retina using the b/a ratio in humans. In dogs, evaluation using the b/a ratio has not been reported, whereas the intensity of SF has been defined. In this study, we performed a convenient ERG recording method using a contact lens electrode with a built-in light source (LED-electrode), and confirmed SF as reported previously. ERG recordings were performed on 15 healthy beagle dogs under sedation. We performed bilateral ERG at 12 different intensities after 30 min dark adaptation. After 10 min light adaptation, we recorded single flash cone and flicker cone response using the SF determined in this study. In this study, SF of 3.0 cd/m(2)/sec (6,000 cd/m(2), 0.5 msec) resulted in b/a=2. The intensity for rod response that recorded only the b-wave was 0.0096 cd/m(2)/sec (80 cd/m(2), 0.12 msec). We could achieve ERG for each response easily and smoothly under sedation, and without general anesthesia. Using an LED-electrode, we could perform more quantitative and reproducible ERG examinations than with traditional methods. We propose that the b/a ratio is the most useful parameter in ERG reporting for evaluating retinal function.     

Emerging technologies for the chemical control of root parasitic weeds

Parasitic plants in the Orobanchaceae family include devastating weed species, such as Striga, Orobanche, and Phelipanche, which parasitize major crops, drastically reduces crop yields and cause economic losses of over a billion US dollars worldwide. Advances in basic research on molecular and cellular processes responsible for parasitic relationships has now achieved steady progress through advances in genome analysis, biochemical analysis and structural biology. On the basis of these advances it is now possible to develop chemicals that control parasitism and reduce agricultural damage. In this review we summarized the recent development of chemicals that can control each step of parasitism from strigolactone biosynthesis in host plants to haustorium formation.     

A New and Simple Technique for the Isolation of Plasma Membrane Lipids from Root-Tips

It has been suggested that plasma membrane (PM) lipids play a major role in aluminum (Al) tolerance; however, no direct investigations have been carried out using PM lipids from root-tips. Here we report a new technique for PM isolation as an alternative to the laborious two-polymer phase partitioning method that is commonly applied, as follows: 1) separation of protoplasts from 1-cm root-tip portions by enzymatic digestion, 2) attachment of the purified protoplasts to glass plates coated with polylysine, 3) preparation of PM ghosts by successive burst of the attached protoplasts using three separate buffer solutions (25 mM PIPES, 5 mM EDTA, and 2 mM MgCl₂, at pH 7.0) with slow stirring for 60 s. The PMs were confirmed to be devoid of organelle membranes by fluorescence microscopy, thin layer chromatography (TLC) and western blot analysis. The PM lipids obtained were found to be useful for studies on their differential permeability and lipid composition between lines of triticale or cultivars of maize under Al stress.     

Correction to: Chlorantraniliprole application differentially affects adult emergence of Sympetrum dragonflies in rice paddy fields

Paddy and Water Environment -     

Residual effects of organic acid-treated phosphate rocks on some soil properties and phosphate availability

The effects of the application of organic acid-treated phosphate rocks on the growth and nutrient uptake of Italian rye grass (Lolium multiflorum Lam. cv. Tachiwase) and some properties of the soil were evaluated in a greenhouse pot experiment. Phosphate rocks (PRs) collected from six countries; China, Florida (USA), Jordan, Sri Lanka, Togo, and Tanzania, were treated with 1 M oxalic or tartaric acid at the ratio of 2.5 mL g^-1 PR. The organic acid-treated PRs, containing 12–31% water soluble P, were applied to a granitic regosol (pH 5.8) at 200 mg P pot^-1 (4 kg soil). Untreated PRs and single superphosphate (SSP) were included in the treatments. Italian ryegrass was grown for 175 dafter planting (DAP) with ample supply of other nutrients and water. Shoots were harvested at 56, 119, and 175 DAP and the soils were analyzed for pH and Olsen-P after the experiment. Application of organic acid-treated PRs consistently increased the dry matter yield and P uptake of the plants compared with the application of untreated PRs at each harvest, but they were less effective than SSP. A larger amount of P (calculated per unit water-soluble P applied) was recovered from the organic acid-treated PRs than from SSP. The amount of residual extractable P in the soils with the organic acid-treated PRs was about the same as or significantly larger than that in the soil treated with SSP. Soil pH was also significantly higher than in the control and SSP soils. The results suggest that organic acids could be used to improve the P availability of PRs to plants with favorable residual effects in terms of available P and soil pH, without exerting any adverse effects on plant growth or nutrient acquisition.