Validation of a sensitive assay for thiocoraline in mouse plasma using liquid chromatography-tandem mass spectrometry

A sensitive high-performance liquid chromatography-tandem mass spectrometry assay for thiocoraline, an anti-tumor depsipeptide, in mouse plasma is described. Echinomycin, a quinoxaline peptide, was used as an internal standard. Thiocoraline was recovered from the mouse plasma using protein precipitation with acetonitrile and followed by solid-phase extraction of the supernatant. The mobile phase consisted of methanol (0.1% formic acid)-water (0.1% formic acid) (90:10, v/v). The analytical column was a YMC C(18). The standard curve was linear from 0.1 to 50 ng/ml (R(2)>0.99). The lower limit of quantitation was 0.1 ng/ml. The assay was specific based on the multiple reaction monitoring transitions at m/z 1157-->215 and m/z 1101-->243 for thiocoraline and the internal standard, echinomycin, respectively. The mean intra- and inter-day assay accuracies remained below 5 and 12%, respectively, for all calibration standards and quality control (QC) samples. The intra- and inter-day assay precisions were less than 11.4 and 9.5% for all QC levels, respectively. The utility of the assay was demonstrated by a pharmacokinetic study of i.v. (bolus) thiocoraline on CD-1 mice. Thiocoraline was stable in mouse plasma in an ice-water bath for 6 h and for three freeze-thaw cycles. The reconstituted thiocoraline after extraction and drying sample process was stable in the autosampler for over 24 h. The assay was able to quantify thiocoraline in plasma up to 48 h following dose. Pharmacokinetic analysis showed that thiocoraline has distinct pharmacokinetic profiling when dosed in different formulation solutions. The assay is currently used to measure thiocoraline plasma concentrations in support of a project to develop a suitable formulation with a desirable pharmacokinetic profile.     

Characterization of TioQ, a type II thioesterase from the thiocoraline biosynthetic cluster

An antitumor agent thiocoraline is a thiodepsipeptide marine product derived from two Micromonospora sp. strains that inhibits protein synthesis by binding of its key 3-hydroxyquinaldic acid (3HQA) chromophores to duplex DNA. There are at least two potential pathways via which the 3HQA moiety could be biosynthesized from L-Trp. By biochemical characterization and by preparation of knockouts of an adenylation-thiolation enzyme, TioK, and of two type II thioesterases, TioP and TioQ, found in the thiocoraline biosynthetic gene cluster, we gained valuable insight into the pathway followed for the production of 3HQA.     

A New Scaffold of an Old Protein Fold Ensures Binding to the Bisintercalator Thiocoraline

Thiocoraline is a thiodepsipeptide with potent antitumor activity. TioX, a protein with an unidentified function, is encoded by a gene of the thiocoraline biosynthetic gene cluster. The crystal structure of the full-length TioX protein at 2.15 Å resolution reveals that TioX protomer shares an ancient βαβββ fold motif with glyoxalase I and bleomycin resistance protein families, despite a very low sequence homology. Intriguingly, four TioX monomers form a unique 2-fold symmetric tetrameric assembly that is stabilized by four intermolecular disulfide bonds formed cyclically between Cys60 and Cys66 of adjacent monomers. The arrangement of two of the four monomers in the TioX tetramer is analogous to that in dimeric bleomycin resistance proteins. This analogy indicates that this novel higher-order structural scaffold of TioX may have evolved to bind thiocoraline. Our equilibrium titration studies demonstrate the binding of a thiocoraline chromophore analog, quinaldic acid, to TioX, thereby substantiating this model. Furthermore, a strain of Streptomyces albus containing an exogenous thiocoraline gene cluster devoid of functional tioX maintains thiocoraline production, albeit with a lower yield. Taken together, these observations rule out a direct enzymatic function of TioX and suggest that TioX is involved in thiocoraline resistance or secretion.     

Interaction of N,N,N-trialkylammonioundecahydro-closo-dodecaborates with dipalmitoyl phosphatidylcholine liposomes

N,N,N-Trialkylammonioundecahydrododecaborates (1-), a novel class of compounds of interest for use as anions in ionic liquids, interact with DPPC liposomes. Increasing compound concentration causes an increasing negative ζ potential. Dissociation constants demonstrate that the binding capacity increases strongly with longer chain length. N,N,N-Trialkylammonioundecahydrododecaborates with longer alkyl chains show a detergent-like behavior: the compounds incorporate into the liposome membrane and differential scanning calorimetric experiment show already low concentrations cause a complete disappearance of the peak representing the gel-to-liquid crystalline phase transition. In contrast, compounds with shorter alkyl chains only interact with the headgroups of the lipids. Investigations by means of cryo-TEM reveal that all derivatives induce significant morphological changes of the liposomes. N,N,N-Trialkylammonioundecahydrododecaborates with short alkyl chains produce large bilayer sheets, whereas those with longer alkyl chains tend to induce the formation of open or multi-layered liposomes. We propose that the binding of N,N,N-trialkylammonioundecahydrododecaborates is mainly due to electrostatic interactions between the doubly negatively charged cluster unit and the positively charged choline headgroup; the positively charged ammonium group might be in contact with the deeper-lying negatively charged phosphate. For N,N,N-trialkylammonioundecahydrododecaborates with longer alkyl chains hydrophobic interactions with the non-polar hydrocarbon part of the membrane constitute an additional important driving force for the association of the compounds to the lipid bilayer.     

Genotypic effects of fertilization on seedling sweetgum biomass allocation,N uptake,and N use efficiency

Screening and selecting tree genotypes that are responsive to N additions and that have high nutrient use efficiencies can provide better genetic material for short-rotation plantation establishment. A pot experiment was conducted to test the hypotheses that (1) sweetgum ( Liquidambar styraciflua L.) families have different patterns in biomass production and allocation, N uptake, and N use efficiency (NUE), because of their differences in growth strategies, and (2) sweetgum families that are more responsive to N additions will also have greater nutrient use efficiencies. Seedlings from two half-sib families (F10022 and F10023) that were known to have contrasting responses to fertility and other stress treatments were used for an experiment with two levels of N (0 vs. 100 kg N/ha equivalent) and two levels of P (0 vs. 50 kg P/ha equivalent) in a split-plot design. Sweetgum seedlings responded to N and P treatments rapidly, with increases in both size and biomass production, and those responses were greater with F10023 than with F10022. Growth response to N application was particularly strong. N and P application increased the proportional allocation of biomass to leaves. Under increased N supply, P application increased foliar N concentration and content, as well as total N uptake by the seedlings. However, NUE was decreased by N addition and was higher in F10023 than in F10022 when P was not limiting. A better understanding of genotype by fertility interactions is important in selecting genotypes for specific site conditions and for optimizing nutrient use in forestry production.     

NA Proteins of Influenza A Viruses H1N1/2009, H5N1, and H9N2 Show Differential Effects on Infection Initiation,Virus Release,and Cell-Cell Fusion

Two surface glycoproteins of influenza virus, haemagglutinin (HA) and neuraminidase (NA), play opposite roles in terms of their interaction with host sialic acid receptors. HA attaches to sialic acid on host cell surface receptors to initiate virus infection while NA removes these sialic acids to facilitate release of progeny virions. This functional opposition requires a balance. To explore what might happen when NA of an influenza virus was replaced by one from another isolate or subtype, in this study, we generated three recombinant influenza A viruses in the background of A/PR/8/34 (PR8) (H1N1) and with NA genes obtained respectively from the 2009 pandemic H1N1 virus, a highly pathogenic avian H5N1 virus, and a lowly pathogenic avian H9N2 virus. These recombinant viruses, rPR8-H1N1NA, rPR8-H5N1NA, and rPR8-H9N2NA, were shown to have similar growth kinetics in cells and pathogenicity in mice. However, much more rPR8-H5N1NA and PR8-wt virions were released from chicken erythrocytes than virions of rPR8-H1N1NA and rPR8-H9N2NA after 1 h. In addition, in MDCK cells, rPR8-H5N1NA and rPR8-H9N2NA infected a higher percentage of cells, and induced cell-cell fusion faster and more extensively than PR8-wt and rPR8-H1N1NA did in the early phase of infection. In conclusion, NA replacement in this study did not affect virus replication kinetics but had different effects on infection initiation, virus release and fusion of infected cells. These phenomena might be partially due to NA proteins’ different specificity to α2-3/2-6-sialylated carbohydrate chains, but the exact mechanism remains to be explored.     

A comparison of snRNP-associated Sm-autoantigens: human N, rat N and human B/B''.

N is a tissue-specific, Sm-epitope bearing, snRNP-associated protein found predominantly in brain. The cDNA sequence encoding human N is compared to those for rat N and human B/B'. The amino acid sequences of human and rat N are 100% conserved. Although the amino acid sequences of N and B/B' are very similar to each other, B/B' contains 50 amino acids which are not present in N. On Northern blots the cDNAs encoding N and B/B' recognize two different RNA species. A comparison of the codon usage, as specified by the open reading frames of N and B/B' as well as results from Southern blots, show that N and B/B' are derived from different genes.     

Pathogenicity of Nocardia caviae,N. asteroides and N. brasiliensis

The pathogenicity ofNocardia caviae, N. asteroides andN. brasiliensis has been tested for white mice, guinea pigs and rabbits and chorio-allantoic membrane of the developing chick embryo. Altogether, 14 strains belonging to the 3Nocardia species originating from soil, human and animal sources in India or abroad were tested. All of them proved pathogenic though the degree of virulence varied from strain to strain. Incorporation of hog gastric mucin in the inoculum enhanced the virulence of all the 3Nocardia species for white mice.N. caviae strains were uniformly more virulent than those ofN. asteroides andN. brasiliensis.In the white mice inoculated intraperitoneally, a greater dissemination of the disease was apparent withN. caviae than withN. asteroides. Of the 6 strains ofN. caviae tested, 5 disseminated to the lung, 3 to the heart and 2 to the brain. InN. asteroides dissemination of the disease to the brain was observed with 2 of its 3 strains.N. brasiliensis showed no dissemination.N. caviae was found to be equally virulent for white mice, guinea pigs and rabbits. On the other hand,N. asteroides andN. brasiliensis were more virulent for white mice than for guinea pigs and rabbits. The lesions caused byN. caviae in mice, guinea pigs and rabbits persisted up to 4 weeks. In strong contrast to this the lesions due toN. asteroides andN. brasiliensis found in the guinea pigs and rabbits showed a strong tendency towards spontaneous clearance.Histologically, the lesions caused byN. caviae, N. asteroides andN. brasiliensis in mice, guinea pigs and rabbits were in the form of abscesses which showed an acute or chronic reaction. In the case ofN. caviae these abscesses showed both granules and freely dispersed cocco-bacillary bodies or filaments. As forN. asteroides it occurred in the form of cocco-bacillary bodies or filaments whereasN. brasiliensis consistently produced granules in the lesions.The lesions caused by the 3Nocardia species on the chorio-allantoic membrane of the developing chick embryo were in the form of abscesses which contained cocco-bacillary bodies and branching filaments but no granules.This forms a part of the thesis submitted by P.V.K. for Ph. D. degree, of the University of Delhi.     

麻栎种源林叶片碳、氮、磷化学计量特征的变异

结合林木育种方法,以3个麻栎种源试验林为研究对象,测定了29个麻栎种源叶片碳、氮、磷化学计量特征.结果表明: 地点(环境)显著影响叶片碳、氮、磷、碳氮比、碳磷比、氮磷比,解释量占总变异量的13.2%～66.7%,而种源(遗传)的影响并不显著,解释量只占总变异量的2.9%～11.0%.叶片氮与碳氮比、氮与氮磷比、磷与碳磷比、磷与氮磷比均具有显著的相关性,且无论是地点间还是东西部种源间均存在共同的标准化主轴斜率.表明在单一树种(麻栎)水平上,种源间具有相似的碳、氮、磷生化过程,其叶片化学计量特征主要受环境的影响;而化学计量特征在地点间和东西部种源间稳定的相关系数反映出叶片化学计量特征的特定耦合比例不受环境和种源的影响,支持化学计量学的内稳性理论.     

水肥互作对滴灌玉米氮素吸收、水氮利用效率及产量的影响

优化水、氮供应是实现作物高产与水肥资源高效利用的有效途径.本文研究了田间试验条件下,水(4500、6750、9000 m³·hm^-2)、氮(0、225、330、435、540 kg·hm^-2)互作对高密度(≥105000 株·hm^-2)滴灌玉米干物质积累、氮素吸收及产量的影响.结果表明: 玉米干物质积累与吸氮量均随灌溉和施氮水平的增加明显升高,当施氮量大于435 kg·hm^-2和灌溉量大于9000 m³·hm^-2时则呈减少趋势.完熟期玉米干物质积累对灌水的响应表现为W₆₇₅₀(36359 kg·hm^-2)>W₉₀₀₀(35077 kg·hm^-2)>W₄₅₀₀(33451 kg·hm^-2),施氮对玉米吸氮量的变化表现为N₄₃₅(459.9 kg·hm^-2)>N₅₄₀(458.1 kg·hm^-2)>N₃₃₀(416.3 kg·hm^-2)>N₂₂₅(351.3 kg·hm^-2),N₄₃₅比N₃₃₀、N₂₂₀分别升高9.1%、32.7%,N₅₄₀比N₄₃₅降低0.6%.在施氮量0～435 kg·hm^-2范围内,玉米最大氮素吸收速率随施氮量增加而升高,在施氮量为435 kg·hm^-2时达最大(6.57 kg·hm^-2·d^-1).灌水与施氮均可显著增加玉米产量、穗粒数和穗粒质量,二者有明显的正交互作用,且以氮为主效应.在施氮0～435 kg·hm^-2范围内,氮肥利用率随施氮量的增加而升高,此后反而降低;灌溉水分生产率随施氮量升高而增加,随灌水量增加而明显下降,灌溉定额为4500～6750 m³·hm^-2时,灌溉水分生产率可达2.57～3.80 kg·m^-3.玉米最高产量18072 kg·hm^-2的施氮量为567.0 kg·hm^-2.最佳经济施氮量为427.9～467.7 kg N·hm^-2时,玉米产量在17109～17138 kg·hm^-2,氮素偏生产力和氮肥利用率分别达122 kg N·hm^-2和45.0%.水氮一体化施肥可实现滴灌玉米高产协同水、氮利用效率的共同提高.     

N,N,N-三甲基壳聚糖甲基硫酸盐的合成及理化性质的测定

壳聚糖与甲醛、甲酸反应得到N,N-二甲基壳聚糖,然后以硫酸二甲酯为季铵化试剂反应得到N,N,N-三甲基壳聚糖甲基硫酸盐(TMCMS),用IR1、H NMR和元素分析对其结构进行了表征。元素分析结果表明其季铵化度为74.6%,差示扫描量热法和热重分析法结果表明其热稳定性比壳聚糖差,但其水溶性明显优于壳聚糖,25℃时在水中的溶解度可达20 mg/mL,浓度为2 mg/mL时在pH 3～12范围内无沉淀产生。     

Autumnal N storage determines the spring growth,N uptake and N internal cycling of young peach trees

Although N storage determines early spring growth in trees, the usefulness of autumn N supply remains unclear as N uptake decreases in autumn, but could be restored earlier in spring to compensate for low N cycling. We intended here to evaluate the effects of autumn N supply on N uptake, storage and cycling, and spring growth. Four levels of N fertilisation were applied to 1-year-old peach trees, between the end of shoot growth and leaf fall. In spring, N supply was ¹⁵N labelled. Organ dry weights and concentrations of ¹⁴N, ¹⁵N, starch and soluble sugars were evaluated after the first growth flush. Bud development had previously been described in the same trees by Jordan et al. (Trees-Struct Func 23:235–245, 2009). Fertilisation promoted autumn N uptake, spring N uptake and growth up to a threshold level, since no differences were evidenced between the three highest N treatments. The variability in tree ¹⁴N contents was related to the number of phytomers per tree in autumn, i.e. to tree size. In spring, the depletion of the perennial structures was independent of treatment, indicating a complete mobilisation of the N stores. Spring growth was related to the amounts of cycling N, and spring N uptake was in turn proportioned to shoot and fruit growth. The lower N uptake of the N limited trees was not due to a C shortage since these trees displayed the highest starch concentrations. We conclude that a moderate autumn fertilisation improved spring growth and fruit production (Jordan et al. in Trees-Struct Func 23:235–245, 2009) and that a deficit of N storage could not be compensated for by an increase in spring N uptake.     

Electrosprayed polyelectrolyte complexes between mucoadhesive N,N,N,-trimethylchitosan-homocysteine thiolactone and alginate/carrageenan for camptothecin delivery

Novel hydrogel polyelectrolyte complexes (PECs) between the N,N,N,-trimethylchitosan-homocysteine thiolactone (TM-HT-chitosan) and two anionic polymers were investigated. The particles of pure thiolated chitosan and its PECs with alginate and carrageenan were fabricated using the electrospray ionization technique. The hydrogel PEC particles were characterized by scanning electron microscopy, dynamic light scattering, Fourier transform infrared microscopy, thermogravimetric analysis, encapsulation efficiency (EE), mucoadhesive property and in vitro drug release behavior. TM-HT-chitosan/alginate particles could be loaded with camptothecin (CPT), employed as a model anti-cancer drug, at an over 70% EE, and revealed both a reduced burst effect and a prolonged release of CPT over 3 days. The resultant TM-HT-chitosan/alginate PEC particles displayed a 5.60-, 1.86- and 1.55-fold stronger mucoadhesive property compared to that of the unmodified chitosan/alginate PEC at pH 1.2, 4.0 and 6.4, respectively, and this was not affected by the CPT loading level.     

Lipoxygenase-mediated hydrogen peroxide-dependent N-demethylation of N,N-dimethylaniline and related compounds

To date, studies of xenobiotic N-demethylation have focused on heme-proteins such as P450 and peroxidases. In this study we investigated the ability of non-heme iron proteins, namely soybean lipoxygenase (SLO) and human term placental lipoxygenase (HTPLO) to mediate N-demethylation of N,N-dimethylaniline (DMA) and related compounds in the presence of hydrogen peroxide. In addition to being hydrogen peroxide dependent, the reaction was also dependent on incubation time, concentration of enzyme and DMA and the pH of the medium. Using Nash reagent to estimate formaldehyde production, we determined the specific activity for SLO mediated N-demethylation of DMA to be 200 + 18 nmol HCHO/min per mg protein or 23 +/- 2 nmol/min per nmol of enzyme, while that of HTPLO was 33 +/- 4 nmol HCHO/min per mg protein. Nordihydroguaiaretic acid (NDGA), a classical inhibitor of lipoxygenase (LO), as well as antioxidants and free radical reducing agents, caused a marked reduction in the rate of production of formaldehyde from DMA by SLO. Besides N,N-dimethylaniline, N-methylaniline, N,N,N',N'-tetramethylbenzidine, N,N-dimethyl-p-phenylenediamine, N,N-dimethyl-3-nitroaniline and N,N-dimethyl-p-toluidine were also demethylated by SLO. The formation of a DMA N-oxide was not detected. Preliminary experiments suggested SLO-mediated hydrogen peroxide-dependent S-dealkylation of methiocarb or O-dealkylation of 4-nitroanisole does not occur.     

Comparative analyses of pandemic H1N1 and seasonal H1N1, H3N2, and influenza B infections depict distinct clinical pictures in ferrets

Influenza A and B infections are a worldwide health concern to both humans and animals. High genetic evolution rates of the influenza virus allow the constant emergence of new strains and cause illness variation. Since human influenza infections are often complicated by secondary factors such as age and underlying medical conditions, strain or subtype specific clinical features are difficult to assess. Here we infected ferrets with 13 currently circulating influenza strains (including strains of pandemic 2009 H1N1 [H1N1pdm] and seasonal A/H1N1, A/H3N2, and B viruses). The clinical parameters were measured daily for 14 days in stable environmental conditions to compare clinical characteristics. We found that H1N1pdm strains had a more severe physiological impact than all season strains where pandemic A/California/07/2009 was the most clinically pathogenic pandemic strain. The most serious illness among seasonal A/H1N1 and A/H3N2 groups was caused by A/Solomon Islands/03/2006 and A/Perth/16/2009, respectively. Among the 13 studied strains, B/Hubei-Wujiagang/158/2009 presented the mildest clinical symptoms. We have also discovered that disease severity (by clinical illness and histopathology) correlated with influenza specific antibody response but not viral replication in the upper respiratory tract. H1N1pdm induced the highest and most rapid antibody response followed by seasonal A/H3N2, seasonal A/H1N1 and seasonal influenza B (with B/Hubei-Wujiagang/158/2009 inducing the weakest response). Our study is the first to compare the clinical features of multiple circulating influenza strains in ferrets. These findings will help to characterize the clinical pictures of specific influenza strains as well as give insights into the development and administration of appropriate influenza therapeutics.     

Anti N1 Cross-Protecting Antibodies Against H5N1 Detected in H1N1 Infected People

The A(H5N1) influenza virus pandemic may be the result of avian H5N1 adapting to humans, leading to massive human to human transmission in a context of a lack of pre-existing immunity. As A(H1N1) and A(H5N1) share the same neuraminidase subtype, anti-N1 antibodies subsequent to H1N1 infections or vaccinations may confer some protection against A(H5N1). We analysed, by microneutralization assay, the A/Vietnam/1194/04 (H5N1) anti-N1 cross-protection acquired either during A/NewCaledonia/20/99 (H1N1) infection or vaccination. In cases with documented H1N1 infection, H5N1 cross-protection could be observed only in patients born between 1930 and 1950. No such protection was detected in the sera of vaccinated individuals.     

Interactions between perennial ryegrass ( Lolium perenne L.) and white clover (Trifolium repens L.) under contrasting nitrogen availability: productivity,seasonal patterns of species composition,N2 fixation,N transfer and N recovery

Nitrogen (N) fertiliser and clover cultivar choice affect competition and productivity in grass-clover mixtures. Pure stands and mixtures of perennial ryegrass and white clover cultivars with contrasting growth habits were examined. The aim of this work was to study the effect of repetitive nitrogen (N) application and cultivar combination on competition and productivity, N yield in the harvested herbage, N₂ fixation in mixtures and pure stands, and transfer of N from clover to the companion grass. Large-leaved white clover cultivar Alice and small-leaved cv. Gwenda and perennial ryegrass cvs. Barlet (erect) and Heraut (prostrate) were sown in pure stands and as four binary grass-clover mixtures on a sandy soil in 1995. In the mixtures, two levels of N fertiliser were applied: 0 (-N) and 150 and 180 kg ha^-1 y^-1 N (+N) in 1996 and 1997, respectively, while the grass monocultures received three N levels (0, 140/180 and 280/360 kg ha^-1) in 1996 and 1997, respectively. No N was applied to pure clover. The plots were cut five times during 1996 and six times during 1997. Fertiliser N was applied in early spring and after every harvest. The treatments were continued until the summer of 1999. In pure grass, the applied N was effectively recovered. In mixtures, N application affected competition by enhancing grass growth and the overall effect of N application was 17 kg DM per kg N applied in 1996. However, there was no yield response to N fertilizer in 1997, because this was compensated for by a higher clover production in unfertilised mixtures. In 1997, -N mixtures yielded more N than +N mixtures, owing to the higher clover content and N₂ fixation. Large-leaved clover cv. Alice was better able to withstand the negative effect of repetitive N application on clover production in mixtures and increased its proportion during the growing season of the second harvest year. In 1997, mixtures with Alice yielded more N than mixtures with Gwenda, but in pure clover swards, there was no cultivar effect on N yield. Also, during the autumn of 1998 and the spring of 1999, the clover content was highest in mixtures with Alice. Harvested N and apparent N₂ fixation were almost twice as high in 1997 as in 1996. N yield and apparent N₂ fixation were higher in pure clover than in mixtures. In mixtures, the apparent N₂ fixation in 1996 was 142 kg N ha^-1, irrespective of cultivar or N treatment. In 1997, it was on average 337 kg N ha^-1, and higher in -N mixtures and in mixtures with Alice. For each tonne of clover DM in the harvested herbage, 65 and 57 kg N was harvested in 1996 and 1997 in -N mixtures, respectively. The apparent transfer of clover-derived N to grass was on average 29 and 70 kg N ha^-1 yr^-1 in 1996 and 1997, respectively. It was highest in +N mixtures and highest in mixtures with Gwenda in 1997. In contrast to clover, the grass cultivars were very similar in their productivity and seasonal patterns, despite their contrasting growth habits. Seasonal trends in N yield, N transfer and N recovery are discussed in relation to fertilizer application regimes and variation in production patterns in mixtures and pure stands. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of N,K, and P fertilization,N source,and clipping on potential tetany hazard of bromegrass

Summary The objective of this field study was to determine early-season effects of N source, N, K, and P fertilization, and clipping (to simulate grazing) on potential tetany hazard of bromegrass (Bromus inermis L.) as indicated by the chemical composition of its forage. Tetany is a metabolic disorder of ruminants resulting from forage with low Mg availability. Chemical components considered in the forage were inorganic cations, organic acids, aconitate, and per cent total N/per cent total water soluble carbohydrate (N/TWSC). Differences between the sum (in meq/kg) of inorganic cations (Mg, Ca, K, and Na) and inorganic anions (Cl, NO₃, H₂PO₄, and SO₄) in forage were defined as the concentration of organic acids (C-A). Soil was Parshall fsl, a Pachic Haploboroll. Yields and chemical composition of oven-dried forage from previously unclipped and reclipped plots were determined at 3-week intervals beginning May 22 and June 12, respectively. A water budget was determined using soil-water and rainfall data.Forage yields were increased 2- to 3-fold by N fertilization with the NO₃-N source generally outyielding the NH₄-N source. A slight additional yield response to that obtained with N alone was obtained with K+P fertilization but not with K or P alone with or without N. Much less total forage was removed from reclipped plots than from unclipped plots. Forage Mg content was decreased only slightly by K or NH₄-N fertilization. Soil analysis indicated that high NH₄-N levels were present at the May 22 harvest. Magnesium and Ca concentrations were only slightly affected by N fertilization; however, K, K/(Ca+Mg), total N, C-A, and aconitate were increased. Reclipping increased Mg, N, K, N/TWSC, C-A, and aconitate. Estimates of blood-plasma Mg concentrations were obtained by using the data for plant N, K, and Mg. These estimates did not indicate increased tetany hazard as a result of reclipping, but did indicate increased tetany hazard from N fertilization. Forage C-A and aconitate concentrations were decreased by fertilization with KCl which seemed to have been caused by the increased Cl concentrations in the forage. Estimates of quantities of Mg, arriving at the root surfaces from the soil by mass flow, far exceeded amounts of Mg in the forage. Mass flow seemed to be the principal mechanism by which Mg and Ca arrived at root surfaces but this mechanism was much less important for K.This study indicated an increased potential tetany hazard resulting primarily from N fertilization with either NH₄-N or NO₃-N sources. However, the potential for increased forage and livestock-carrying capacity with N fertilization is very large. Therefore, management practices corroborated by livestock data are vitally needed to minimize tetany hazard while increasing bromegrass yields by N fertilization.Contribution from Soil, Water, and Air Sciences, North Central and Northeastern Regions, ARS-USDA.Follett, Power, and Grunes are soil scientists and Kleinis a biological laboratory technician. Follett is now National Program Staff Scientist, ARS, BARC-West, Beltsville, MD 20705. Power and Klein are at the USDA Northern Great Plains Research Center, Mandan, ND 58554, as formerly was Follett. Grunes is at the U.S. Plant, Soil and Nutrition Laboratory, Ithaca, NY 14853.     

沙地植被碳氮磷化学计量特征与物种多样性的关系

植被化学计量特征表征植物营养限制.它是否会影响物种多样性需要进一步探究.本研究以宁夏哈巴湖国家级自然保护区沙地油蒿群落和沙柳群落为对象,计算了植被碳(C)、氮(N)、磷(P)含量,分析了植被C、N、P生态化学计量特征对沙地植物群落物种多样性的影响.结果表明: 在流动沙丘和半固定沙丘生境中的沙柳群落,物种多样性Simpson指数与植被C/N值存在显著的负相关关系,而与植被N/P值的相关性不显著;在半固定沙丘和固定沙丘生境中的油蒿群落,物种多样性Shannon指数与N/P值存在显著正相关关系,而与C/N值存在显著负相关关系.结合C、N、P生态化学计量特征与冗余分析可知,油蒿群落和沙柳群落的植被P含量对物种多样性的影响不同,导致两个群落的植被N/P值对物种多样性产生不同的影响.