A sustainable phosphorus management in agriculture: Assessing trade-offs between human health risks and nutritional yield regarding heavy metals in maize grain.

High-quality products in sustainable agriculture require both limited health risks and sufficient dietary nutrients. Phosphorus (P) as a finite and non-renewable resource is widely used in agriculture, usually exerting influence on the accumulation of heavy metals (HMs) in soil and crops. The present research explores, for the first time, the combined effects of long-term P fertilizer and repeated zinc (Zn) application in field on the human health risks and nutritional yield regarding trace elements in maize grain. A field experiment was conducted using maize with six P application rates (0, 12.5, 25, 50, 100, and 200 kg P ha-1) and two Zn application rates (0 and 11.4 kg Zn ha-1). The results showed that the concentrations of Zn, copper (Cu), and lead (Pb) in the maize grain were significantly affected by P application and can be further affected by Zn application. The concentrations of chromium (Cr) and arsenic (As) showed opposite tendency as affected by P fertilizer rates while did not affected by additional Zn application. Zn application decreased the cadmium (Cd) concentration at high P levels and Pb concentration at low P levels, particularly. No HMs contamination or direct health risk was found in maize grain after receiving long-term P and repeated Zn fertilizer. The threshold hazard quotient of an individual and all investigated HMs in this study were acceptable for human digestion of maize grain. While the carcinogenic risk of Cr was non-negligible in case of maize was taken as one of daily staple food for local residents. Combination use of P (25 kg ha-1) and Zn fertilizer on maize enhanced its nutritional supply ability regarding Zn and Cu, and simultaneously mitigated potential human health risks associated with Cd and Pb.

[Chemical constituents from Clausena excavata and their inhibitory activities against proliferation of synoviocytes].

The chemical constituents from the stems and leaves of Clausena excavata were isolated and purified by column chromatography with silica gel, ODS, Sephadex LH-20 and RP-HPLC. The chemical structures of the isolated compounds were identified on the basis of physicochemical properties, spectroscopic analysis, as well as the comparisons with the data reported in literature. Nineteen compounds were isolated from the 90% ethanol extract of the stems and leaves of C. excavata, which were identified as methyl orsellinate(1), syringaresinol(2), lenisin A(3), scopoletin(4), osthenol(5), N-benzoyltyrarnine methyl ether(6), N-p-coumaroyltyramine(7), aurantiamide acetate(8), 1H-indole-3-carboxaldehyde(9), furostifoline(10), clausenalansine E(11), 3-formylcarbazole(12), clausine L(13), clausine E(14), methyl carbazole-3-carboxylate(15), glycosinin(16), murrayafoline A(17), clausine H(18) and 2,7-dihydroxy-3-formyl-1-(3'-methyl-2'-butenyl)carbazole(19). Among these isolated compounds, compounds 1-11 were isolated from C. excavata for the first time, and compounds 1, 2 and 10 were isolated from the genus Clausena for the first time. In addition, this study evaluated the anti-rheumatoid arthritis activities of compounds 1-19 by measuring their anti-proliferative effects on synoviocytes in vitro according to MTS method. Compounds 10-19 displayed remarkable anti-rheumatoid arthritis activities, which exhibited the inhibitory effects on the proliferation of MH7 A synovial fibroblast cells with the IC_(50) values ranging from(27.63±0.18) to(235.67±2.16) µmol·L~(-1).

Foliar Zinc Application to Wheat May Lessen the Zinc Deficiency Burden in Rural Quzhou, China.

Zinc (Zn) malnutrition is a common health problem, especially in developing countries. The human health and economic benefits of the replacement of conventional flour with Zn-biofortified wheat flour in rural household diets were assessed. One hundred forty-five wheat flour samples were collected from rural households in Quzhou County. Then, field experiments were conducted on wheat at two Zn levels (0 and 0.4% ZnSO4 · 7H2O foliar application) under 16 diverse agricultural practices in Quzhou County. Foliar Zn application significantly increased the Zn concentration and bioavailability in wheat grain and flour. If rural households consumed Zn-biofortified flour instead of self-cultivated flour or flour purchased from supermarkets, 257-769 or 280-838, 0.46-1.36 million or 0.50-1.49 million disability-adjusted life years (DALYs) lost, respectively, could be saved in Quzhou County and China. Amounts of 2.3-12.0 million and 5.5-22.6 billion RMB could be obtained via Zn-biofortified flour in Quzhou County and China, respectively. The current study indicates that Zn-biofortified flour via foliar Zn application is a win-win strategy to maintain the yield and combat human Zn deficiency in rural households in China. More health and economic benefits could be obtained in rural household dependent on wheat flour purchased from supermarkets than in those dependent on self-cultivated wheat flour.

Responses in Zinc Uptake of Different Mycorrhizal and Non-mycorrhizal Crops to Varied Levels of Phosphorus and Zinc Applications.

Negative effects of high phosphorus (P) application on zinc (Zn) nutrition have been observed in many crops. This study investigated the Zn responses of three typical crops to varied P and Zn applications. A pot experiment was conducted using two mycorrhizal crops (maize and soybean) and one non-mycorrhizal crop (oilseed rape) under three levels of P, two levels of Zn, and two levels of benomyl. Results showed that P application significantly decreased shoot and root Zn concentrations, Zn uptake, and Zn acquisition efficiency (ZnAE) of the three crops irrespective of Zn rate, and that these reductions were greater for maize and soybean than for oilseed rape. Zn application alleviated the P inhibition of Zn uptake in the three crops. The arbuscular mycorrhizal fungi (AMF) colonization of maize and soybean contributed most to the negative effects of increasing P application on Zn uptake, explaining 79-89 and 64-69% of the effect, respectively. For oilseed rape, root dry weight and root Zn concentration explained 90% of the decrease in Zn uptake caused by P application. These results suggest that there is another pathway in addition to the mycorrhizal pathway regulating Zn uptake under mediation by P supply.

The responses of soil enzyme activities, microbial biomass and microbial community structure to nine years of varied zinc application rates.

Zinc (Zn) fertilizer application can certainly improve the production and nutritional quality of cereal crops. However, Zn accumulation in the soil may lead to some deleterious environmental impacts in agroecosystems. The effects of long-term Zn application on soil microbial properties remain unclear, but it is imperative to understand such effects. In this study, we collected soil samples from a nine-year field experiment in a wheat-maize system that continuously received Zn applied at various rates (0, 2.3, 5.7, 11.4, 22.7 and 34.1 kg ha-1) to evaluate the soil enzymes, microbial biomass and microbial community structure. The results showed that Zn application at the rate of 5.7 kg ha-1 significantly increased the activities of urease, invertase, alkaline phosphatase and catalase in the soil, while the rate of 34.1 kg ha-1 significantly decreased the evaluated enzyme activities. The microbial biomass carbon (C) and nitrogen (N) were not affected by Zn application rates, although an increase in the microbial biomass C was observed in the 11.4 kg ha-1 treatment. Moreover, the alpha diversity of the bacterial and fungal communities did not vary among the nil Zn, optimal Zn (5.7 kg ha-1) and excess Zn (34.1 kg ha-1) treatments. However, the bacterial communities in the soil receiving the optimal and excess Zn application rates were slightly changed. Compared to the nil Zn treatment, the other Zn application rates increased the relative abundances of the Rhodospirillales, Gaiellales and Frankiales orders and decreased the abundance of the Latescibacteria phylum. The redundancy analysis further indicated that the soil bacterial community composition significantly correlated with the concentrations of soil DTPA-Zn and total Zn. These results highlight the importance of optimal Zn application in achieving high production and high grain quality while concurrently promoting soil microbial activity, improving the bacterial community and further maintaining the sustainability of the agroecological environment.

Health risk assessment associated with heavy metal accumulation in wheat after long-term phosphorus fertilizer application.

Phosphorus (P) fertilizer is widely used to increase wheat yield. However, it remains unclear whether prolonged intake of wheat grain that received long-term P application may promote human health risks by influencing heavy metal(loid)s (HMs) accumulation. A 10-year field experiment was conducted to evaluate the effects of continuous P application (0, 25, 50, 100, 200, and 400 kg P ha-1) on human health risks of HMs, including zinc (Zn), copper (Cu), cadmium (Cd), lead (Pb), arsenic (As), nickel (Ni), and chromium (Cr), by ingesting wheat grain. The results showed that P application facilitated Zn, Pb, Cd, and As accumulation in the topsoil. The Zn, Cu, Pb, and Ni concentrations in grain were decreased, while Cd and As were increased by P application. All HMs concentrations of both soil and grain were in the ranges of corresponding safety thresholds at different P levels. The accumulation abilities of Zn, Cu, Pb, and Ni from soil and straw to grain were suppressed by P addition while of As was enhanced. There was no significant difference in the hazard index (HI) of the investigated HMs in all treatments except 25 kg ha-1. The threshold cancer risk (TCR) associated with As and Cd was enhanced, while that of Pb was alleviated as P application increased. Behaviors of Cr from soil to wheat and to humans were not affected by P application. Phosphorus application at a rate of 50 kg ha-1 decreased total non-cancer and cancer risks by 15% and 21%, respectively, for both children and adults, compared to the highest value. In conclusion, long-term optimal application of 50 kg P ha-1 to wheat did not result in additional adverse effects on the total non-carcinogenic or carcinogenic risk caused by the studied HMs to humans through the ingestion of wheat grain.

Molecularly imprinted solid phase extraction method for simultaneous determination of seven nitroimidazoles from honey by HPLC-MS/MS.

In this work, a selective sample cleanup procedure that combined molecular imprinting technique with solid phase extraction was developed for the simultaneous extraction of the seven nitroimidazoles (NMZs) from honey samples. The molecular imprinting polymers for NMZs were prepared through bulk polymerization method using 2-methyl-5-nitroimidazole as template molecule, methacrylic acid as the functional monomer and ethylene glycol dimethacrylate as the cross linking agent. The obtained molecular imprinting polymers showed high affinity to template molecule and was used as selective sorbent for simultaneously selective extraction of the seven NMZs from honey matrix. An off-line molecularly imprinted solid phase extraction (MISPE) method followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) for simultaneous determination of the seven NMZs from honey samples was also established. The proposed method was validated at 1.0, 2.0 and 10.0µg/kg, obtaining recoveries in the range of 79.7-110%, with repeatability and interday precision values (expressed as relative standard deviation) ≤11.4% and ≤15.2%, respectively. Limits of quantification for different NMZs were 1.0µg/kg, which were always below the minimum required performance limits established by the European Community Reference Laboratories (Commission Decision 2002/657/EC). It was demonstrated that this proposed MISPE-HPLC-MS-MS method could be applied to direct determination of NMZs from honey samples.

[Analysis of conserved flanking elements associated with antibiotic resistance genes dissemination].

The overuse of antibiotics in medicine, animal husbandry, and aquiculture industry increases the emergence of antibiotic resistant bacteria and antibiotic resistance genes (ARGs), and also, accelerates the dissemination of ARGs within environmental bacteria. In this study, the total DNA was directly extracted from environmental samples, and the upstream and downstream of antibiotic resistance genes were directly amplified by thermal asymmetric interlaced PCR (Tail-PCR) technique. By optimizing the Tail-PCR program, the multiple flanking sequences of tetW, including 6 upstream sequences and 9 downstream sequences, were simultaneously acquired. Through the bioinformatics analysis, the upstream of tetW presented a perfect inverted repeat (IR), a known tetW regulator peptide, and an insertional sequence (IS), whereas the downstream of tetW presented a most conservative fragment and a common open reading frame (ORF) coding methyltransferase. This study not only revealed several conserved flanking tetW gene modules, but also supplied a highly-efficient and convenient methodology for the research of tetW's dissemination within bacteria, i. e., several flanking sequences could be concisely obtained from one sample by using Tail-PCR program.

Genetic diversity and variance of Stentor coeruleus (Ciliophora: Heterotrichea) inferred from inter-simple sequence repeat (ISSR) fingerprinting.

We used inter-simple sequence repeat fingerprinting to analyze the genetic structure of 16 populations of Stentor coeruleus from three lakes and three ponds in China. Using 14 polymorphic primers, a total of 99 discernible DNA fragments were detected, among which 76 (76.77%) were polymorphic, indicating median genetic diversity in these populations. Further, both Nei's gene diversity (h) and Shannon's information index (I) between the different populations revealed a median genetic diversity. At the same time, gene flow was interpreted to be low. The main factors responsible for the median level of diversity and low gene flow within populations are probably due to a low frequency of sexual recombinations. Analysis of molecular variance showed that there was high genetic differentiation among the five water bodies. Both cluster analysis and a nonmetric multidimensional scaling analysis suggested that genotypes isolated from the same locations displayed a higher genetic similarity than those from different ones, separating populations into subgroups according to their geographical locations. However, there is a weak positive correlation between the genetic distance and geographical distance.

[Biological process of phosphorus turnover in surface water body of Xiamen Harbor. II: Grazing pressure of copepod on phytoplankton].

To understand the roles of copepod in the biogeochemical cycling of phosphorus, gut fluorescence method was applied to examine in situ the grazing rate of copepod on the phytoplankton in Xiamen Time Station (XMTS) in May, August and November 2005 and March 2006. In the meanwhile, the abundance and species composition of copepod were investigated, and the grazing pressure of copepod on the phytoplankton was estimated. The results showed that the annual average grazing rate of copepod was 55.53 microg x m(-3) x d(-1), being the highest (108.98 microg x m(-3) x d(-1)) in autumn and the lowest (7.18 microg x m(-3) x d(-1)) in summer. Based on the estimation from our experimental data, the daily grazing rate of copepod populations on the phytoplankton in Xiamen Harbor was, on annual average, about 1.81% of the phytoplankton's standing stock, with the values in spring, summer, autumn, and winter being 3.22%, 0.06%, 3.52% and 0.46%, respectively.

Molecular profiling of hepatocellular carcinomas developing spontaneously in acyl-CoA oxidase deficient mice: comparison with liver tumors induced in wild-type mice by a peroxisome proliferator and a genotoxic carcinogen.

By using cDNA microarrays, we studied the expression profiles of 26 hepatocellular carcinomas (HCC) developing spontaneously in peroxisomal fatty acyl-CoA oxidase null (AOX-/-) mice. The development of liver tumors in AOX-/- mice is due to sustained activation of peroxisome proliferator-activated receptor alpha (PPARalpha) by the unmetabolized substrates of AOX, which serve as natural PPARalpha ligands. We then compared the AOX-/- liver tumor expression profiles with those induced by ciprofibrate, a non-genotoxic peroxisome proliferator, or by the genotoxic carcinogen diethylnitrosamine (DENA) to discern differences in gene expression patterns that may predict or distinguish PPARalpha-mediated liver tumors from genotoxically derived tumors. Our results show that HCCs developing in AOX-/- mice share a number of deregulated (up- or down-regulated) genes with ciprofibrate-induced liver tumors. The overall commonality of expression between AOX-/- and ciprofibrate-induced liver tumors but not with DENA-induced tumors strongly implicates the activation of PPARalpha and PPARalpha-regulated genes in liver, including those participating in lipid catabolism, as key factors in the development of HCC in AOX-/- and in ciprofibrate-treated mice. Northern blot analysis confirmed the differential expression of some of the genes identified in the present study, and also some genes identified previously as PPARalpha regulated, such as CD36, lymphocyte antigen 6 complex locus (Ly-6D), and C3f. We found a panel of 12 genes upregulated in all three classes of liver tumors, namely AOX-/-, ciprofibrate-induced and DENA-induced. These include an uncharacterized RIKEN cDNA, lipocalin 2, insulin-like growth factor-binding protein 1, Ly-6D and CD63 among others. In conclusion, these results identify distinguishing features between non-genotoxic and genotoxic carcinogen derived liver tumors as well as genes that are upregulated in both types and suggest that RIKEN cDNA, Ly-6D and lipocalin 2 in particular appear to be desirable molecular markers for further study in liver carcinogenesis and progression.

Adipocyte-specific gene expression and adipogenic steatosis in the mouse liver due to peroxisome proliferator-activated receptor gamma1 (PPARgamma1) overexpression.

Peroxisome proliferator activated-receptor (PPAR) isoforms, alpha and gamma, function as important coregulators of energy (lipid) homeostasis. PPARalpha regulates fatty acid oxidation primarily in liver and to a lesser extent in adipose tissue, whereas PPARgamma serves as a key regulator of adipocyte differentiation and lipid storage. Of the two PPARgamma isoforms, PPARgamma1 and PPARgamma2 generated by alternative splicing, PPARgamma1 isoform is expressed in liver and other tissues, whereas PPARgamma2 isoform is expressed exclusively in adipose tissue where it regulates adipogenesis and lipogenesis. Since the function of PPARgamma1 in liver is not clear, we have, in this study, investigated the biological impact of overexpression of PPARgamma1 in mouse liver. Adenovirus-PPARgamma1 injected into the tail vein induced hepatic steatosis in PPARalpha(-/-) mice. Northern blotting and gene expression profiling results showed that adipocyte-specific genes and lipogenesis-related genes are highly induced in PPARalpha(-/-) livers with PPARgamma1 overexpression. These include adipsin, adiponectin, aP2, caveolin-1, fasting-induced adipose factor, fat-specific gene 27 (FSP27), CD36, Delta(9) desaturase, and malic enzyme among others, implying adipogenic transformation of hepatocytes. Of interest is that hepatic steatosis per se, induced either by feeding a diet deficient in choline or developing in fasted PPARalpha(-/-) mice, failed to induce the expression of these PPARgamma-regulated adipogenesis-related genes in steatotic liver. These results suggest that a high level of PPARgamma in mouse liver is sufficient for the induction of adipogenic transformation of hepatocytes with adipose tissue-specific gene expression and lipid accumulation. We conclude that excess PPARgamma activity can lead to the development of a novel type of adipogenic hepatic steatosis.

Expression of peroxisome proliferator-activated receptor alpha, and PPARalpha regulated genes in spontaneously developed hepatocellular carcinomas in fatty acyl-CoA oxidase null mice.

Fatty acyl-CoA oxidase null mice (AOX-/-) develop hepatocellular carcinomas in 100% of animals between 10 and 15 months. We evaluated spontaneously developed HCC in AOX-/- mice for PPARalpha, PPARalpha regulated genes and peroxisome volume density and compared with adjacent non-neoplastic liver and liver in wild-type (AOX+/+) and heterozygous (AOX+/-) mice. The level of PPARalpha mRNA was 2.5-fold higher in HCC compared to the adjacent liver. mRNAs of PPARalpha regulated genes such as peroxisomal bifunctional enzyme, thiolase, cytochrome P450 CYP4A1 and CYP4A3 were similar in HCC and adjacent liver and increased by 7- to 22-fold compared with wild-type and heterozygous mice. Immunoblot analysis of HCC showed high amounts of PPARalpha, peroxisomal bifunctional enzyme and thiolase. Electron microscopic examination revealed 3.8 and 8.3-fold increase in the volume density of peroxisomes in HCC and adjacent liver, respectively, compared to the volume density in wild-type mice. These results demonstrate that spontaneously developed HCC in AOX-/- mice display a similar type of pleiotropic responses to high levels of PPARalpha ligands as the non-neoplastic liver. The changes observed in HCC and adjacent liver in AOX-/- mice were identical to those observed in rats and mice exposed to peroxisome proliferators.

Interaction of PIMT with transcriptional coactivators CBP, p300, and PBP differential role in transcriptional regulation.

PIMT (PRIP-interacting protein with methyltransferase domain), an RNA-binding protein with a methyltransferase domain capable of binding S-adenosylmethionine, has been shown previously to interact with nuclear receptor coactivator PRIP (peroxisome proliferator-activated receptor (PPAR)-interacting protein) and enhance its coactivator function. We now report that PIMT strongly interacts with transcriptional coactivators, CBP, p300, and PBP but not with SRC-1 and PGC-1alpha under in vitro and in vivo conditions. The PIMT binding sites on CBP and p300 are located in the cysteine-histidine-rich C/H1 and C/H3 domains, and the PIMT binding site on PBP is in the region encompassing amino acids 1101-1560. The N-terminal of PIMT (residues 1-369) containing the RNA binding domain interacts with both C/H1 and C/H3 domains of CBP and p300 and with the C-terminal portion of PBP that encompasses amino acids 1371-1560. The C-terminal of PIMT (residues 611-852), which binds S-adenosyl-l-methionine, interacts respectively with the C/H3 domain of CBP/p300 and with a region encompassing amino acids 1101-1370 of PBP. Immunoprecipitation data showed that PIMT forms a complex in vivo with CBP, p300, PBP, and PRIP. PIMT appeared to be co-localized in the nucleus with CBP, p300, and PBP. PIMT enhanced PBP-mediated transcriptional activity of the PPARgamma, as it did for PRIP, indicating synergism between PIMT and PBP. In contrast, PIMT functioned as a repressor of CBP/p300-mediated transactivation of PPARgamma. Based on these observations, we suggest that PIMT bridges the CBP/p300-anchored coactivator complex with the PBP-anchored coactivator complex but differentially modulates coactivator function such that inhibition of the CBP/p300 effect may be designed to enhance the activity of PBP and PRIP.