Detoxification of the post-harvest antifungal pesticide thiabendazole by cold atmospheric plasma.

Cold atmospheric plasma (CAP) irradiation has a sterilizing effect without thermal denaturation or the production of residual substances. Hence, it is considered to be a safe sterilization technology with minimal damage for fresh foods. In addition, its decomposition effect on chemical substances has also been confirmed, and the application of CAP in the food and agricultural domains is increasing. In this study, we examined the potential of CAP to detoxify pesticide residues. Post-harvest chemical treatments using pesticides, such as fungicides, are frequently employed in imported agricultural products and are often disapproved by consumers. Therefore, we assessed the detoxification of thiabendazole (TBZ), a widely used post-harvest pesticide, using low-cost air plasma irradiation. We found that CAP irradiation conditions that detoxified TBZ caused little damage to the edible parts of mandarin oranges. The results of the present study suggest that CAP irradiation is useful for detoxifying and degrading pesticide residues without damaging agricultural products and that CAP irradiation is an effective means of maintaining food safety.

Crystal structures of aconitase X enzymes from bacteria and archaea provide insights into the molecular evolution of the aconitase superfamily.

Aconitase superfamily members catalyze the homologous isomerization of specific substrates by sequential dehydration and hydration and contain a [4Fe-4S] cluster. However, monomeric and heterodimeric types of function unknown aconitase X (AcnX) have recently been characterized as a cis-3-hydroxy-L-proline dehydratase (AcnXType-I) and mevalonate 5-phosphate dehydratase (AcnXType-II), respectively. We herein elucidated the crystal structures of AcnXType-I from Agrobacterium tumefaciens (AtAcnX) and AcnXType-II from Thermococcus kodakarensis (TkAcnX) without a ligand and in complex with substrates. AtAcnX and TkAcnX contained the [2Fe-2S] and [3Fe-4S] clusters, respectively, conforming to UV and EPR spectroscopy analyses. The binding sites of the [Fe-S] cluster and substrate were clearlydifferent from those that were completely conserved in other aconitase enzymes; however, theoverall structural frameworks and locations of active sites were partially similar to each other.These results provide novel insights into the evolutionary scenario of the aconitase superfamilybased on the recruitment hypothesis.

Enzymatically polymerised polyphenols prepared from various precursors potentiate antigen-specific immune responses in both mucosal and systemic compartments in mice.

Despite significant modern medicine progress, having an infectious disease is a major risk factor for humans. Mucosal vaccination is now widely considered as the most promising strategy to defeat infectious diseases; however, only live-attenuated and inactivated mucosal vaccines are used in the clinical field. To date, no subunit mucosal vaccine was approved mainly because of the lack of safe and effective methodologies to either activate or initiate host mucosal immune responses. We have recently elucidated that intranasal administration of enzymatically polymerised caffeic acid potentiates antigen-specific mucosal and systemic antibody responses in mice. However, our earlier study has not confirmed whether these effects are specific to the polymer synthesised from caffeic acid. Here, we show that enzymatically polymerised polyphenols (EPPs) from various phenolic compounds possess mucosal adjuvant activities when administered nasally with an antigen to mice. Potentiation of antigen-specific immune responses by all EPPs tested in this study showed no clear difference among the precursors used. We found that intranasal administration of ovalbumin as the antigen, in combination with all enzymatically polymerised polyphenols used in this study, induced ovalbumin-specific mucosal IgA in the nasal cavity, bronchoalveolar lavage fluid, vaginal fluids, and systemic IgG, especially IgG1, in sera. Our results demonstrate that the mucosal adjuvant activities of polyphenols are not limited to polymerised caffeic acid but are broadly observable across the studied polyphenols. These properties of polyphenols may be advantageous for the development of safe and effective nasal vaccine systems to prevent and/or treat various infectious diseases.

[Two Cases of Breast Cancer with Extensive Invasion to Skin].

Case 1: A 48-year-old female was admitted to our hospital because of left HER2 type breast cancer with a skin invasion of 10×11 cm in the chest wall. Since she had previously received anti-HER2 therapy, we performed anti-HER2 therapy in our department as well, but the tumor gradually became larger and presented with a cancerous ulcer. The use of metronidazole gel in the tumor area reduced the odor. The tumor progressed and she died 1 year and 1 month after the first visit to our hospital. Case 2: A 51-year-old female visited our hospital because of a cauliflower-like right breast tumor measuring 20×17 cm with bleeding and infection. After diagnosis of right breast cancer with multiple bone metastasis, CMF therapy was performed, and then 40 cycles of docetaxel and bevacizumab therapy were performed. As a result, her breast tumor is no longer visible macroscopically, and she runs her daily life without problems. Breast cancer with an extensive skin invasion has a poor prognosis. However, in some cases such as case 2, the proper chemotherapy might be beneficial for long survival.

Novel non-phosphorylative pathway of pentose metabolism from bacteria.

Pentoses, including D-xylose, L-arabinose, and D-arabinose, are generally phosphorylated to D-xylulose 5-phosphate in bacteria and fungi. However, in non-phosphorylative pathways analogous to the Entner-Dodoroff pathway in bacteria and archaea, such pentoses can be converted to pyruvate and glycolaldehyde (Route I) or α-ketoglutarate (Route II) via a 2-keto-3-deoxypentonate (KDP) intermediate. Putative gene clusters related to these metabolic pathways were identified on the genome of Herbaspirillum huttiense IAM 15032 using a bioinformatic analysis. The biochemical characterization of C785_RS13685, one of the components encoded to D-arabinonate dehydratase, differed from the known acid-sugar dehydratases. The biochemical characterization of the remaining components and a genetic expression analysis revealed that D- and L-KDP were converted not only to α-ketoglutarate, but also pyruvate and glycolate through the participation of dehydrogenase and hydrolase (Route III). Further analyses revealed that the Route II pathway of D-arabinose metabolism was not evolutionally related to the analogous pathway from archaea.

Anesthetic effect of a combination of medetomidine-midazolam-butorphanol in cynomolgus monkeys (Macaca fascicularis).

The anesthetic effect of a combination of medetomidine, midazolam and butorphanol (Me-Mi-Bu) was evaluated in healthy cynomolgus monkeys. The Me-Mi-Bu combination was intramuscularly administered as follows: Dose 1, Me 0.015 mg/kg-Mi 0.1 mg/kg-Bu 0.15 mg/kg; Dose 2, Me 0.02 mg/kg-Mi 0.15 mg/kg-Bu 0.2 mg/kg; and Dose 3, Me 0.04 mg/kg-Mi 0.3 mg/kg-Bu 0.4 mg/kg. The combination rapidly induced immobilization, and lateral recumbency was reached within 15 min. The duration of anesthesia for each dose administered was follows: Dose 1, 47 ± 27 min; Dose 2, 113 ± 31 min; and Dose 3, 190 ± 24 min. The anesthetic effect of the combination was abolished by the α2-adrenoceptor antagonist atipamezole. No marked changes in the levels of hematologic or serum biochemical parameters were noted in cynomolgus monkeys administered the combination plus atipamezole. Taken together, these results suggest that the Me-Mi-Bu combination exhibits reversible anesthetic effect and may be useful for studies involving cynomolgus monkeys.

Effects of transport stress on serum alkaline phosphatase activity in beagle dogs.

Here, to determine the effects of transport stress on blood parameters in dogs, we investigated the changes in hematologic and serum chemical parameters in healthy beagle dogs transported from Beijing, China, to Osaka, Japan, to obtain the background data. Only the activity of serum alkaline phosphatase increased clearly upon arrival, a change attributed to transport stress, but the activity gradually reduced afterward. No marked changes in levels of other blood parameters were noted. Our findings here suggest that alkaline phosphatase is a useful tool for studying transport stress.

Ability of water-soluble biosubstances to eliminate hydroxyl and superoxide radicals examined by spin-trapping ESR measurements: two-dimensional presentation of antioxidative ability.

The hydroxyl- and superoxide-radical-eliminating ability of water-soluble biosubstances was examined by ESR combined with the spin-trapping method, indicating a median inhibitory dose, ID(h)(50) (mM) and id(h)(50) (mg/mL) for the hydroxyl radical, and ID(s)(50) (mM) and id(s)(50) (mg/mL) for the superoxide radical. Both the 1/[ID(h)(50) (mM)] and 1/[ID(s)(50) (mM)] values of selected biosubstances were linearly related to the second-order rate constant, k(2) (M(-1) s(-1)), defined for the reaction between biosubstances and the radicals in a logarithmic presentation. The result indicates that ID(h)(50) (mM) and ID(s)(50) (mM) are suitable parameters for both types of radical-eliminating ability. The obtained results are depicted two-dimensionally, taking id(h)(50) (mg/mL) as the abscissa and id(s)(50) (mg/mL) as the ordinate in the ROS inhibitory diagram. The biosubstances tested were assigned to five separate areas characterized by their functional groups on the diagram. The obtained ROS inhibitory diagram indicates the possibility for screening appropriate antioxidants.

Immunochromatography for the rapid determination of cadmium concentrations in wheat grain and eggplant.

BACKGROUND: A simple and quick on-site test for trace levels of cadmium (Cd) in food is needed because of the human toxicity of this heavy metal. We developed an immunochromatography kit which uses the antigen-antibody complex reaction between the Cd-ethylenediaminetetraacetic acid (Cd-EDTA) complex and an anti-Cd-EDTA antibody. We previously reported the successful use of this kit to determine Cd concentrations in brown rice with respect to the international standard: 0.4 mg kg⁻¹. Here, we measured, using this immunochromatography kit, Cd concentrations in crops with lower international standards than rice. RESULTS: Cadmium extracted with 0.1 mol L⁻¹ HCl from wheat grain and fresh eggplant was purified sufficiently using an ion-exchange column treatment. Appropriate HCl extraction rates and dilution rates for the column eluate were selected; Cd concentrations in wheat grain and fresh eggplant were determined successfully by immunochromatography with respect to the international standards of 0.2 mg kg⁻¹ and 0.05 mg kg⁻¹ fresh weight, respectively. CONCLUSION: Approximate Cd concentrations in wheat grain and fresh eggplant can be monitored easily and quickly by this method at locations where facilities for acid digestion and precision analysis are not available.

Retention behavior on aminoethyl-modified poly(p-phenylene terephthalamide) fiber stationary phases in gas chromatography.

Surface derivatization of Kevlar, poly(p-phenylene terephthalamide), fiber has been studied along with the evaluation of the surface characteristics of the chemically-modified fiber as the stationary phase in packed-capillary gas chromatography (GC). Several experimental parameters in the derivatization reaction have been optimized, and the retention behavior of the surface-derivatized fibrous stationary phase has been investigated using various standard solutes, such as alkanes, alcohols and alkylbenzenes. By introducing aminoethyl functional groups onto the surface of the fibrous material, a specific selectivity for polar solutes has been observed.

Restoration of cadmium-contaminated paddy soils by washing with ferric chloride: Cd extraction mechanism and bench-scale verification.

The ability of FeCl3 to extract Cd from three paddy soils was compared with that of various irons, manganese, and zinc salts to elucidate the extraction mechanism. Manganese, zinc and iron salts (including FeCl3) extracted 4-41%, 8-44% and 24-66% of total Cd, respectively. This difference reflected the pH of the extraction solution, indicating that the primary mechanism of Cd extraction by FeCl3 is proton release coupled with hydroxide generation, as iron hydroxides are insoluble. Washing with FeCl3 led to the formation of Cd-chloride complexes, enhancing Cd extraction from the soils. FeCl3 effectively extracted Cd from all of the three soils compared to HCl that is a conventional washing chemical, when the concentrations of the two washing chemicals were between 15 and 60mM(c) (at above extraction pH 2.4), while the corresponding extraction pH of FeCl3 was slightly higher than HCl. As HCl is the strong acid of complete dissociation, if excess amount of HCl was added to soil, it is possible to give the dissolution of clay minerals in soils. In contrast, proton release from FeCl3 is controlled by the chemical equilibrium of hydroxide formation. While soil fertility properties were affected by a bench-scale soil washing with 45mM(c) FeCl3, adverse effects were not crucial and could be corrected. The bench-scale test confirmed the effectiveness of FeCl3 for removal of soil Cd. The washing had no negative effect on rice yield and lowered the Cd concentration of rice grain and rice straw in a pot experiment.

Remediation of cadmium contamination in paddy soils by washing with chemicals: selection of washing chemicals.

The efficiencies of neutral salts, strong acids, and chelates were tested for extracting cadmium (Cd) from three paddy soils. The higher the selectivity of the cations of the added neutral salts toward soil adsorption sites, the lower the pH in the extracts and the more soil Cd could be extracted. In addition, soil carbon and nitrogen contents and mineral composition were closely associated with the amount of Cd extracted. Calcium chloride and iron(III) chloride were selected as wash chemicals to restore Cd-contaminated paddy soils in situ. Washing with calcium chloride led to the formation of Cd chloride complexes, enhancing Cd extraction from the soils. The washing also substantially decreased soil levels of exchangeable and acid-soluble Cd, which are the major forms of bioavailable Cd for rice (Oryza sativa L.). The optimum conditions for in situ soil washing were also determined for calcium chloride.

Identification of a putative voltage-gated Ca2+ -permeable channel (OsTPC1) involved in Ca2+ influx and regulation of growth and development in rice.

Cytosolic free Ca2+ serves as an important second messenger participating in signal transduction of various environmental stresses. However, molecular bases for the plasma membrane Ca2+ influx and its regulation remain largely unknown. We here identified a gene (OsTPC1) encoding a putative voltage-gated Ca2+ channel from rice, ubiquitously expressed in mature leaves, shoots and roots as well as in cultured cells. OsTPC1 rescued the Ca2+ uptake activity and growth rate of a yeast mutant cch1. To elucidate its physiological roles, we generated transgenic rice plants and cultured cells overexpressing OsTPC1 mRNA. Furthermore, a retrotransposon (Tos17) insertional knockout mutant of OsTPC1 was isolated. OsTPC1-overexpressing cells showed hypersensitivity to excess Ca2+ but higher growth rate under Ca2+ limitation, while growth of the OsTPC1-knockout cultured cells was less sensitive to extracellular free Ca2+ concentration, suggesting that OsTPC1 has Ca2+ transport activity across the plasma membrane. OsTPC1-overexpressing plants showed reduced growth and abnormal greening of roots. Growth of Ostpc1 seedlings was comparable to the control on agar plates, while significantly reduced in adult plants. These results suggest that OsTPC1 functions as a Ca2+ -permeable channel involved in the regulation of growth and development.