15N-NMR spectroscopy. IV. Comparison of poly(L-lysine) and isopoly(L-lysine)

The natural abundance ¹⁵N-nmr spectroscopy has been used to characterize the isomeric polymers (L -Lys)_n and iso (L -Lys)_n in aqueous solution. Although the peptide nitrogens of the two polymers have nearly equivalent shifts at pH < 10, the amino nitrogens differ by 5–6 ppm at pH < 7 and provide an easy means of identification. Furthermore, the polymers are distinguishable by the pK_a of the amino group and the basicity of the peptide nitrogen. At pH 10.3 and 25°C, (Lys)_n exhibits line broadening and an upfield chemical shift of the peptide nitrogen, indicative of the coil → helix transition. The formation of 100% helix may produce a shift as large as 5 ppm, which probably makes ¹⁵N-nmr spectroscopy more suitable for studies of this transition.     

Development and application of aromatic [13C, 1H] SOFAST-HMQC NMR experiment for nucleic acids

Higher sensitivity of NMR spectrometers and novel isotopic labeling schemes have ushered the development of rapid data acquisition methodologies, improving the time resolution with which NMR data can be acquired. For nucleic acids, longitudinal relaxation optimization in conjunction with Ernst angle excitation (SOFAST-HMQC) for imino protons, in addition to rendering rapid pulsing, has been demonstrated to yield significant improvements in sensitivity per unit time. Extending such methodology to other spins offers a viable prospect to measure additional chemical shifts, thereby broadening their utilization for various applications. Here, we introduce the 2D [¹³C, ¹H] aromatic SOFAST-HMQC that results in overall sensitivity gain of 1.4- to 1.7-fold relative to the conventional HMQC and can also be extended to yield long-range heteronuclear chemical shifts such as the adenine imino nitrogens N1, N3, N7 and N9. The applications of these experiments range from monitoring real-time biochemical processes, drug/ligand screening, and to collecting data at very low sample concentration and/or in cases where isotopic enrichment cannot be achieved.     

Synthesis of Pyrrolo[2,3-d]pyrimidines that are Structurally Related to Methylated Guanosines from tRNA and the Nucleoside Q Analogs,PreQ0 and PreQ1

Abstract

The N-3- and N-2-methylated analogs of several 5-substituted 2 amino-7-(β-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidin-4-ones were synthesized from 5-cyano-2,4-dichloro-7-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (10). These compounds are analogs of nucleoside Q that are methylated in a manner similar to some of the naturally occurring methylated guanosines.

     

Using the dual isotope method to assess cecal amino acid absorption of goat whey protein in rats,a pilot study

Measurement of ileal amino acids (AA) bioavailability is recommended to evaluate protein quality. A dual isotope tracer method, based on plasma isotopic enrichment ratios, has been proposed to determine true digestibility in humans. In a pilot study, we aimed to evaluate whether this method could be implemented in rats to determine AA bioavailability based on isotopic enrichment ratios measured in cecal digesta or plasma samples. Goat milk proteins were intrinsically labeled with ¹⁵N and ²H. Wistar rats were fed a meal containing the doubly labeled goat whey proteins and a tracer dose of ¹³C-spirulina. Blood samples were collected 0, 1 h and 3 h after meal ingestion from the tail vein. The rats were euthanized 4 h (n?=?6) or 6 h (n?=?6) after meal to collect plasma and intestinal contents. True orocecal protein digestibility and AA bioavailability were assessed by means of ¹⁵N and ²H enrichment in cecum content and compared with absorption indexes determined at the plasma or cecum level using isotopic ratios. Plasma kinetics of isotopic enrichment could not be completed due to the limited quantity of plasma obtained with sequential blood collection. However, the absorption indexes determined from cecal ¹⁵N or ²H/¹³C ratios gave coherent values with true orocecal AA bioavailability. This dual isotope approach with measurements of isotopic ratios in digestive content could be an interesting strategy to determine true AA bioavailability in ileal digesta of rats.

     

Theoretical study of isotope effects on the stereodynamics of H<Superscript>+</Superscript>+HD and its isotopic variant D<Superscript>+</Superscript>+HD

The stereodynamics for H⁺+HD and its isotopic variant D⁺+HD were studied with a quasi-classical trajectory (QCT) method at a collision energy of 0.7 eV on the ground 1¹A′ potential energy surface (PES). The polarization-dependent differential cross-sections (PDDCSs) in the center-of-mass frame are presented here. Furthermore, the distribution of the angle between k and j′, p(θ _r ) and the distribution of the dihedral angle p(ϕ _r ) were calculated and are discussed. The results indicate that isotopic substitution exerts substantial effects on the differential cross-section and the product’s rotational polarization.     

Lipid biomarker and carbon isotopic signatures for stromatolite-forming, microbial mat communities and Phormidium cultures from Yellowstone National Park

The molecular and isotopic compositions of lipid biomarkers from cultured filamentous cyanobacteria (Phormidium, also known as Leptolyngbya) have been used to investigate the community and trophic structure of photosynthetic mats from alkaline hot springs of the Lower Geyser Basin at Yellowstone National Park. We studied a shallow‐water coniform mat from Octopus Spring (OS) and a submerged, tufted mat from Fountain Paint Pots (FPP) and found that 2‐methylhopanepolyols and mid‐chain branched methylalkanes were diagnostic for cyanobacteria, whereas abundant wax esters were representative of the green non‐sulphur bacterial population. The biomarker composition of cultured Phormidium‐isolates varied, but was generally representative of the bulk mat composition. The carbon isotopic fractionation for biomass relative to dissolved inorganic carbon (DIC; ?_CO2) for cultures grown with 1% CO₂ ranged from 21.4 to 26.1 and was attenuated by diffusion limitation associated with filament aggregation (i.e. cell clumping). Isotopic differences between biomass and lipid biomarkers, and between lipid classes, depended on the cyanobacterial strain, but was positively correlated with overall fractionation. Acetogenic lipids (alkanes and fatty acids) were generally more depleted than isoprenoids (phytol and hopanoids). The δ¹³C_TOC for OS and FPP mats were somewhat heavier than for cultures (?16.9 and ?23.6, respectively), which presumably reflects the lower availability of DIC in the natural environment. The isotopic dispersions among cyanobacterial biomarkers, biomass and DIC reflected those established for culture experiments. The 7‐methyl‐ and 7,11‐dimethylheptadecanes were from 9 to 11 depleted relative to the bulk organic carbon, whereas 2‐methylhopanols derived from the oxidation‐reduction of bacteriohopanepolyol were enriched relative to branched alkanes by approximately 5–7. These isotopic relationships survived with depth and indicated that the relatively heavy isotopic composition of the OS mat resulted from diffusion limitation. This study supports the suggestion that culture studies can establish valid isotopic relationships for interpretation of trophic structure in modern and ancient microbial ecosystems.     

Production, isolation and characterization of [Leu4]- and [Ile4]surfactins from Bacillus subtilis

Summary Bacillus subtilis coproduces several surfactin variants that are powerful biosurfactants and have potential applications in biology and industry. A single amino acid substitution in the heptapeptide moiety of surfactins strongly modifies their properties. To better establish structure-activity relationships and to search new variants with enhanced properties, Bacillus subtilis was grown into two modified culture media. Two new variants were isolated by chromatographic methods and studied by NMR spectroscopy. As planned, modifications consisted in the substitution of the l-valine residue at the fourth position by a more hydrophobic residue, i.e., leucine or isoleucine. These [Leu⁴]- and [Ile⁴]surfactins have a higher affinity for hydrophobic solvents and a twice improved surfactant power. Structure-property correlations were confirmed by analysis of the hydrophobic residue distribution in the three-dimensional model of the structure of surfactin in solution.Abbreviations DMSO dimethylsulfoxide - 1D, 2D, 3D one-, two-, three-dimensional - TLC thin layer chromatography - CMC critical micellar concentration - NMR nuclear magnetic resonance     

Seasonal Stable Isotope Records of Otoliths from Ocean-pen Reared and Wild Cod, Gadus morhua

Otoliths of ocean-pen reared cod, Gadus morhua, provide a unique opportunity to examine the lifetime history of the fish. Here we report 18 analyses of such otoliths on seasonal (winter and summer) stable oxygen and carbon isotope ratios. The calculated isotopic temperatures from otoliths of reared cod were essentially in agreement with the temperature record during rearing, suggesting that temperature is a dominant factor in the precipitation of otolith aragonite. Carbon isotope ratios increased with age and leveled off at 4 years old, presumed to correlate with sexual maturity of cod. As compared with otoliths of wild-caught cod, significant differences were found in isotope variation and the correlation between ¹³C and ¹⁸O. These differences were probably attributable to the different environmental constraints and food supply.     

Reduced stomatal responses to light, carbon dioxide and abscisic acid in the presence of sodium ions

Abstract Responses of stomata to light and CO₂ were smaller when detached epidermis of Commelina communis L. was incubated on a medium containing 50 mol m^?3 NaCl than when an equimolar KCl solution was used. Although opening in the light in the absence of CO₂ seemed to be the same whichever salt was present, apertures on KCl solutions were smaller in the dark or with CO₂-containing air. The response to 10^?7 mol dm^?3 ABA was similarly reduced in the presence of NaCl. If there is an optimal NaCl concentration for stomatal CO₂ and light responses it is at or below 25 mol m^?3. These findings point towards control of stomatal movements by light, CO₂ and ABA at the level of cation uptake or extrusion.     

Dietary and isotopic specialization: the isotopic niche of three <Emphasis Type="Italic">Cinclodes</Emphasis> ovenbirds

By comparing the isotopic composition of tissues deposited at different times, we can identify individuals that shift diets over time and individuals with constant diets. We define an individual as an isotopic specialist if tissues deposited at different times have similar isotopic composition. If tissues deposited at different times differ in isotopic composition we define an individual as an isotopic generalist. Individuals can be dietary generalists but isotopic specialists if they feed on the same resource mixture at all times. We assessed the degree of isotopic and dietary specialization in three related Chilean bird species that occupy coastal and/or freshwater environments: Cinclodes oustaleti, Cinclodes patagonicus, and Cinclodes nigrofumosus. C. oustaleti individuals were both isotopic and dietary generalists. Tissues deposited in winter (liver and muscle) had distinct stable C (δ¹³C) and stable N isotope ratio (δ¹⁵N) values from tissues deposited in the summer (wing feathers) suggesting that birds changed the resources that they used seasonally from freshwater habitats in the summer to coastal habitats in the winter. Although the magnitude of seasonal isotopic change was high, the direction of isotopic change varied little among individuals. C. patagonicus included both isotopic specialists and generalists, as well as dietary specialists and generalists. The isotopic composition of the feathers and liver of some C. patagonicus individuals was similar, whereas that of others differed. In C. patagonicus, there were large inter-individual differences in the magnitude and the direction of seasonal isotopic change. All individuals of C. nigrofumosus were both isotopic and dietary specialists. The distribution of δ¹³C and δ¹⁵N values overlapped broadly among tissues and clustered in a small, and distinctly intertidal, region of δ space. Assessing individual specialization and unraveling the factors that influence it, have been key questions in animal ecology for decades. Stable isotope analyses of several tissues in appropriate study systems provide an unparalleled opportunity to answer them.     

Comparative evaluation of 99mTc-labeled aminothiols as possible brain perfusion imaging agents

Several new ^99mTc aminodithiols were prepared and evaluated comparatively in experimental animals. The ligands were diamine, triamine or tetramine dithiols. Substituents were either attached on one of the nitrogens or introduced in between the two nitrogens of diamino dithiol (DADT) backbone. ^99mTc-derivatives prepared by coupling DADT to secondary amines via ethylene group showed in mice high initial brain uptake and significant retention in brain tissue. These preparations were mixtures of more than one ^99mTc-complex differing in brain uptake and clearance from the brain. The highest brain retention (brain to blood ratio 2.53, 15 min p.i.) was achieved with the ^99mTc-complex prepared by coupling DADT with ethylene pyrrolidine. Lengthening the chain between the nitrogens of DADT moiety by introducing methyl or amino alkyl groups resulted in ^99mTc-complexes with poor brain accumulation.     

Spatial,ontogenetic and interspecific variability in stable isotope ratios of nitrogen and carbon of Merluccius capensis and Merluccius paradoxus off South Africa

General linear models (GLMs) were used to determine the relative importance of interspecific, ontogenetic and spatial effects in explaining variability in stable isotope ratios of nitrogen (δ¹⁵N) and carbon (δ¹³C) of the co‐occurring Cape hakes Merluccius capensis and Merluccius paradoxus off South Africa. Significant GLMs were derived for both isotopes, explaining 74 and 56% of observed variance in Merluccius spp. δ¹⁵N and δ¹³C, respectively. Spatial effects (west or south coast) contributed most towards explaining variability in the δ¹⁵N model, with Merluccius spp. off the west coast having higher (by c. 1·4‰) δ¹⁵N levels than Merluccius spp. off the south coast. Fish size and species were also significant in explaining variability in δ¹⁵N, with both species showing significant linear increases in δ¹⁵N with size and M. capensis having higher (by c. 0·7‰) δ¹⁵N values than M. paradoxus. Species and coast explained most and similar amounts of variability in the δ¹³C model, with M. capensis having higher (by c. 0·8‰) δ¹³C values than M. paradoxus, and values being lower (by c. 0·7‰) for fishes off the west coast compared with the south coast. These results not only corroborate the knowledge of Merluccius spp. feeding ecology gained from dietary studies, in particular the ontogenetic change in trophic level corresponding to a changing diet, but also that M. capensis feeds at a slightly higher trophic level than M. paradoxus. The spatial difference in Merluccius spp. δ¹⁵N appears due to a difference in isotopic baseline, and not as a result of Merluccius spp. feeding higher in the food web off the west than the south coast, and provides new evidence that corroborates previous observations of biogeographic differences in isotopic baselines around the South African coast. This study also provides quantitative data on the relative trophic level and trophic width of Cape hakes over a large size range that can be used in ecosystem models of the southern Benguela.     

Production of alloplasmic and euplasmic wheat-barley ditelosomic substitution lines 7H<Superscript>1</Superscript>L<Superscript>mar</Superscript>(7D) and analysis of the 18S/5S mitochondrial repeat in these lines

Alloplasmic lines of common wheat with disomic substitution of chromosome 7D for telocentric chromosome 7H¹L^mar of barley H. marinum subsp. gussoneanum Hudson were isolated from the plants of generation BC₃, produced as a result of backcrossing of barley-wheat hybrids H. marinum subsp. gussoneanum (2n = 28) × T. aestivum (2n = 42), Pyrotrix, cultivar, with 28 common wheat cultivars Pyrotrix 28 and Novosibirskaya 67. Chromosome substitution pattern was determined using SSR analysis and C-banding. In preliminary genomic in situ hybridization experiments, telocentric chromosomes were assigned to wild barley was established. In the BC₃F₈ generations of three alloplasmic lines with the 7H¹L^mar(7D) substitution type the differences in fertility manifestation were observed: most of the L-32(1) plants were sterile, in line L-32(2) only sporadic plants were sterile, and line L-32(3) was fertile. Simultaneously with these experiments, using selfpollinated progeny of the hybrids obtained in crosses of common wheat cultivar Saratovskaya 29 (2n = 41), monosomic for chromosome 7D, with common wheat cultivar Pyrotrix 28 with addition of pair of telocentric chromosomes 7H¹L^mar (7D) of barley H. marinum subsp. gussoneanum, euplasmic wheat-barley ditelosomic substitution 7H¹L^mar (7D) lines were isolated. The lines obtained had normal fertility. PCR analysis of the 18S/5S mitochondrial repeat (hereafter, mtDNA sequence) in alloplasmic and euplasmic ditelosomic substitution lines 7H¹L^mar(7D) was performed. In the plants from alloplasmic sterile line L-32(1), the sequences only of the barley (maternal) type were revealed, while the plants from alloplasmic fertile lines L-32(2) and L-32(3) demonstrated heteroplasmy (the presence of barley- and wheat-like sequences within one individual). In euplasmic ditelosomic substitution lines the presence of only wheat-like 18S/5S mitochondrial repeat sequences was observed. The results indicate that the presence of barley-like mtDNA sequences in alloplasmic substitution lines was not associated with the presence of barley chromosomes in their nuclear genomes.     

1H, 13C and 15N NMR assignments and secondary structure of the paramagnetic form of rat cytochrome b 5

Summary Modern multidimensional double- and triple-resonance NMR methods have been applied to assign the backbone and side-chain ¹³C resonances for both equilibrium conformers of the paramagnetic form of rat liver microsomal cytochrome b ₅. The assignment of backbone ¹³C resonances was used to confirm previous ¹H and ¹⁵N resonance assignments [Guiles, R.D. et al. (1993) Biochemistry, 32, 8329–8340]. On the basis of short- and medium-range NOEs and backbone ¹³C chemical shifts, the solution secondary structure of rat cytochrome b ₅ has been determined. The striking similarity of backbone ¹³C resonances for both equilibrium forms strongly suggests that the secondary structures of the two isomers are virtually identical. It has been found that the ¹³C chemical shifts of both backbone and side-chain atoms are relatively insensitive to paramagnetic effects. The reliability of such methods in anisotropic paramagnetic systems, where large pseudocontact shifts can be observed, is evaluated through calculations of the magnitude of such shifts.Abbreviations DANTE delays alternating with nutation for tailored excitation - DEAE diethylaminoethyl - DQF-COSY 2D double-quantum-filtered correlation spectroscopy - EDTA ethylenediaminetetraacetic acid - HCCH-TOCSY 3D proton-correlated carbon TOCSY experiment - HMQC 2D heteronuclear multiple-quantum correlation spectroscopy - HNCA 3D triple-resonance experiment correlating amide protons, amide nitrogens and alpha carbons - HNCO 3D triple-resonance experiment correlating amide protons, amide nitrogens and carbonyl carbons - HNCOCA 3D triple-resonance experiment correlating amide protons, amide nitrogens and alpha carbons via carbonyl carbons - HOHAHA 2D homonuclear Hartmann-Hahn spectroscopy - HOHAHA-HMQC 3D HOHAHA relayed HMQC - HSQC 2D heteronuclear single-quantum correlation spectroscopy - IPTG isopropyl thiogalactoside - NOESY 2D nuclear Overhauser enhancement spectroscopy - NOESY-HSQC 3D NOESY relayed HSQC - TOCSY 2D total correlation spectroscopy - TPPI time-proportional phase incrementation - TSP trimethyl silyl propionate     

In vitro characterization of the inhibitor resistance of glutamate-1-semialdehyde aminotransferase from the cyanobacterium Synechococcus PCC630l GR6

Glutamate-l-semialdehyde (GSA) aminotransferase catalyses the final step in the C5 pathway converting glutamate to the tetrapyrrole precursor δ-aminolaevulinic acid. This enzyme is sensitive to gabaculine (2,3-dihydro-3-amino benzoic acid) and to 4-amino-5-fluoropentanoic acid (AFPA), which are irreversible, mechanism-based inhibitors of pyridoxal phosphatedependent enzymes. Spontaneous mutants of Synechococcus PCC6301 resistant to these inhibitors contain altered enzyme that displays corresponding resistance to high concentrations of the inhibitor. The enzyme from strain GR6, resistant to both inhibitors, contains a three-amino-acid deletion at positions 5–7 and a Met²⁴⁸ → Ile substitution. The enzyme from strain K40 resistant to AFPA but not to gabaculine, contains a Ser¹⁶³ → Thr substitution. GSA aminotransferases containing either the deletion or the substitution that are characteristic of the GR6 mutant were produced in Escherichia coli using the expression vector pMalc2. These engineered mutant enzymes were characterized in terms of their catalytic parameters and sensitivities to gabaculine and AFPA. Furthermore, maltose binding protein/aminotransferase fusion proteins were characterized spectrophotometrically to monitor the interaction of bound cofactor with diamino- and dioxocompounds related to the substrate and both inhibitors. Results were compared with those for similarly produced recombinant wild-type, K40 and GR6 GSA aminotransferases. The engineered products with either the N-terminal deletion or the Met²⁴⁸ → Ile substitution displayed catalytic efficiencies that were intermediate between the wild-type and GR6 or K40 enzymes. However, with respect to their absorption spectra, sensitivity to inhibitors and the reactivity of bound cofactor, they were essentially wild-type. These in vitro studies demonstrate that both changes in enzyme structure are necessary to obtain the distinctive properties of the GR6 aminotransferase, including resistance to high concentrations of gabaculine and AFPA.     

Mother–embryo isotope (δ15N, δ13C) fractionation and mercury (Hg) transfer in aplacental deep‐sea sharks

Stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotopic values and total mercury (Hg) concentrations were analysed in muscle and liver of mothers and embryos of two aplacental shark species, Squalus megalops and Centrophorus moluccensis. Embryos of the two species had similar or lower isotopic values than their respective mothers, the only exception being for δ¹³C, which was higher in the liver of C. moluccensis embryos than in their mothers. Hg concentrations were systematically lower in embryos compared with their mothers suggesting a low transfer of this element in muscle and liver.     

Isotopic evidence of dietary variations and weaning practices in the Gaya cemetery at Yeanri,Gimhae, South Korea

Stable carbon and nitrogen isotope analyses were conducted to investigate dietary variation in human skeletons (n = 109) from the Gaya cemetery at Yeanri located near Gimhae City, South Korea. The cemetery contained three distinct grave types dating to 4th–7th century AD. The main purposes of this research were to reconstruct palaeodiet in the Gaya population and to explore correlations between stable isotope compositions and burial types, inferred age, and sex of these individuals. The isotopic data indicate that the people at Yeanri consumed a predominantly C₃‐based terrestrial diet supplemented with freshwater and/or marine resources. The comparison of isotopic results reveals significant differences in δ¹³C values among three adult burial types (wood‐cist coffin: ?18.5 ± 0.5‰, stone‐cist coffin: ?18.1 ± 0.6‰, mausoleum: ?17.8 ± 0.9‰). Males in wood‐cist and stone‐cist coffins have relatively more elevated mean δ¹³C and δ¹⁵N values than females. The isotopic ratios from the two adult age groups (21–40 years and 40–60 years) indicate that there was no significant dietary change in individuals with age. The isotope data from the infants and children suggest the weaning was a gradual process that was completed between 3 and 4 years of age in the Gaya population. This evidence indicates that the dietary variations within the cemetery reflect social status, sex, and childhood consumption patterns. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc.     

15N nmr spectroscopy. I. Polysarcosine and related sequence polypeptides

The natural abundance ¹⁵N nmr spectra of linear polysarcosine (DP = 35) has been recorded in Me₂SO and H₂O solution. Because of cis/trans isomerization at the peptide bond, a broad signal with several splittings was observed. These splittings appear to reflect the influence of three peptide bonds on a single N atom. The ¹⁵N signals from the sequence polypeptides (β-Ala-Sar-Gly)_n and (β-Ala-Sar-D ,L -Ala)_n also show a cis/trans splitting, as well as chemical shifts which are dependent on the peptide sequence. The tertiary nitrogen of the sarcosyl residue has a T₁ relaxation time which is longer than the T₁ for secondary nitrogens of the other amino acids. The nuclear Overhauser effect is also discussed.     

Effects of growth period,plant age and changes in solution aluminium concentrations on aluminium toxicity in wheat

The effects of growth period (time between transplanting and harvesting), plant age at which aluminium (Al) was added to solution, changes in Al concentration, and solution culture techniques (monitoring and adjusting solution Al concentrations thrice weekly or weekly replacement of the solutions) were investigated using a low ionic strength (2.7×10^–3 M) solution culture technique. The wheat (Triticum aestivum L.) cultivars Waalt (Al-tolerant) and Warigal (Al-sensitive), or the near isogenic lines bred from these cultivars (RR for the Al-tolerant line and SS for the Al-sensitive line) were grown. In all experiments and treatments, Al additions were required to maintain the nominal concentration. The decline in solution Al concentrations was partially attributed to formation of an Al-hydroxy-phosphate precipitate with an Al:P molar ratio of 2.8 to 4.0. Increasing the growth period from 14 to 28 days increased Al sensitivity in Warigal but not in Waalt. When plants were exposed to Al for the same time, increasing the age of the plants that Al was added to solution decreased sensitivity to Al. Differential Al tolerance between the two lines was evident when solutions were monitored thrice weekly or replaced weekly. However, the Al concentration required to reduce relative yield by a given amount when the solutions were replaced weekly was about twice that when the solutions were monitored. With a constant growth period of 28 days, increasing solution Al concentrations for 3 or more days resulted in decreased yields at harvest. The exact effect depended on the cultivar, plant part (tops or roots), when solution Al concentrations were increased and the duration of the increase. For example, increasing Al concentrations from 5 M to 20 M for 10 days reduced yield in the RR line by approximately 50% in the tops and 30% in the roots beyond the effect of 5 M but had no effect in the SS line due to yields already being low at 5 M. Adding 10 M Al to solution for 6 days at the beginning of the experiment reduced yield by 25% in the RR line and 50% in the SS line. In contrast, adding 10 M Al for 6 days in the middle of the growth cycle had no effect on the RR line but reduced yield by approximately 25% in the SS line. These results show that growth period, the age of the plants at which Al is added and the technique used (monitored or weekly replacement) all need to be considered when comparing results from different experiments. These results also show that the Al concentrations in solution need to be regularly monitored in long term experiments.     

High-precision iron isotope analysis of whole blood,erythrocytes, and serum in adults

BackgroundIron isotopic composition serves as a biological indicator of Fe metabolism in humans. In the process of Fe metabolism, essential carriers of Fe circulate in the blood and pass through storage organs and intestinal absorptive tissues. This study aimed to establish an analytical method for high-precision Fe isotopic measurement, investigate Fe concentration and isotopic composition in different parts of whole blood, and explore the potential of Fe isotopic composition as an indicator for Fe status within individuals.Analytical methodsA total of 23 clinically healthy Taiwanese adults of Han descent were enrolled randomly and Fe isotopic compositions of their whole blood, erythrocytes, and serum were measured. The Fe isotopic analysis was performed by Neptune Plus multiple-collector inductively coupled plasma mass spectrometry with double-spike technique. The precision and reproducibility of the Fe isotopic analysis were monitored by international biological and geological reference materials.Main findingsHigh-precision Fe isotopic measurements were achieved alongside with high consistency in the isotopic data for well-characterized reference materials. The Fe isotopic signatures of whole blood and erythrocytes were resolvable from that of serum, where both whole blood and erythrocytes contained significantly lighter Fe isotopic compositions compared to the case of serum (P = 0.0296 and P = 0.0004, respectively). The δ^56/54Fe value of the serum sample was 0.2‰ heavier on an average than those of whole blood or erythrocytes. This isotopic fractionation observed in different parts of whole blood may indicate redox processes involved in Fe cycling, e.g. erythrocyte production and Fe transportation. Moreover, the δ^56/54Fe values of whole blood and serum significantly correlated with the hemoglobin level (P = 0.0126 and P = 0.0020, respectively), erythrocyte count (P = 0.0014 and P = 0.0005, respectively), and Mentzer index (P = 0.0055 and P = 0.0011, respectively), suggesting the Fe isotopic composition as an indicator of functional Fe status in healthy adults. The relationships between blood Fe isotopic compositions and relevant biodemographic variables were also examined. While the average Fe concentration of whole blood was significantly higher in males than in females (P = 0.0028), females exhibited a heavier Fe isotopic composition compared to that of males in whole blood (P = 0.0010) and serum (P < 0.0001). A significantly inverse correlation of the whole blood δ^56/54Fe value with body mass index of individuals (P = 0.0095) was also observed.ConclusionThe results presented herein reveal that blood Fe isotopic signature is consequentially linked to baseline erythrocyte parameters in individuals and is significantly affected by the gender and body mass index in the adult population. These findings support the role of Fe isotopic composition as an indicator for the variance of Fe metabolism among adult individuals and populations and warrant further study to elucidate the underlying mechanisms.