Regional differences in the carbon isotopic compositions of teak from two monsoonal regimes of India

Inter and intra-annual carbon isotope compositions (δ¹³C) of several annual growth rings of teak trees from two monsoonal regimes from India were studied and compared with the corresponding oxygen isotopic (δ¹⁸O) variations. In teak from both the regimes, amplitudes of intra-annual δ¹³C were ∼2-3 times lower than that observed in δ¹⁸O. Seasonal cycle with lower δ¹³C values at the middle and higher at ring boundaries was observed for teak from central India, dominated by the southwest monsoon. Positive correlations of intra-annual δ¹³C values with the corresponding δ¹⁸O values of the same rings and with relative humidity (RH) of the concurrent period suggest a dominant role of RH in controlling δ¹³C values of teak from central India. Intra-annual δ¹³C variations of teak from southern India, receiving both the southwest and northeast monsoons, revealed an initial decreasing trend followed by an increasing trend before culminating in depleted ¹³C values at the end of the growing season. No correlation was observed between intra-annual δ¹³C and δ¹⁸O variations of teak trees from southern India. Regional differences in the climatology of δ¹³C of atmospheric CO₂ or the lengths of growing season could be likely reasons for differing intra-annual δ¹³C variations of teak from the two climatic regimes.     

Anomalous segregations at the Sr6 locus for stem rust resistance in wheat

The action of the gene Sr6 for stem rust resistance in wheat is affected by temperature, light, and the particular susceptible parent with which a line carrying Sr6 has been crossed. Two experiments were carried out to determine whether the effect of the susceptible parents was due to modifier genes, the general genetic background, or interallelic interactions. The data indicated that the susceptible parents carried different sr6 alleles that interacted with Sr6, possibly in a paramutation-like process. In the course of the study, a number of anomalous results were obtained that may be due to the action of transposable elements. Received: 18 February 2000 / Accepted: 31 October 2000     

Non-statistical isotope fractionation as a novel “retro-biosynthetic” approach to understanding alkaloid metabolic pathways

During the biosynthesis of natural products, isotope fractionation occurs due to the selectivity of enzymes for the heavier or lighter isotopomers. As only some of the positions in the molecule are implicated in a given reaction mechanism, position-specific fractionation occurs. Thus, the position-specific ¹³C/¹²C ratios in these compounds can be related to their known precursors and to the known isotope effects of enzymes involved in their biosynthesis, or similar reaction mechanisms. This can be accessed by isotope ratio monitoring NMR spectrometry. In this short review, how isotope fractionation occurs and when it is manifest is described. Then, the way that ¹³C NMR spectrometry has been applied to study certain aspects of the biosynthesis of the solanaceous alkaloids S-(−)-nicotine and tropine is outlined. Notably, it is shown how similar isotope fractionation is found in the steps of the pathway to the common intermediate, N-methyl-Δ¹-pyrrolinium, but that in the moieties derived from different origins no such similarity is found, the isotopic composition of these atoms reflecting their specific metabolic ancestry. In a second example, tramadol, it is shown how this technique can be used in retro-biosynthesis to give direction as to what precursors and pathway intermediates are probable. It is shown how the observed fractionation in the site-specific ¹³C/¹²C ratios can be effectively explained by known metabolism and the properties of enzymes proposed for the pathway. Furthermore, it can give indications of possible mechanisms of those enzymes that are as yet to be described for a number of key steps.     

Stable isotope-based statistical tools as ecological indicator of pollution sources in Mediterranean transitional water ecosystems

N-stable isotope analysis of macroalgae has become a popular method for the monitoring of nitrogen pollution in aquatic ecosystems. Basing on changes in their δ¹⁵N, macroalgae have been successfully used as biological traps to intercept nitrogen inputs. As different nitrogen sources differ in their isotopic signature, this technique provides useful information on the origin of pollutants and their extension in the water body. However, isotopic fractionation potentially resulting from microbial nitrogen processing, and indirect isotopic variations due to effects of physicochemical conditions on algal nutrient uptake and metabolism, may affect anthropogenic N isotopic values during transportation and assimilation. This in turn can affect the observed isotopic signature in the algal tissue, inducing isotopic variations not related to the origin of assimilated nitrogen, representing a “background noise” in isotope-based water pollution studies.In this study, we focused on three neighbouring coastal lakes (Caprolace, Fogliano and Sabaudia lakes) located south of Rome (Italy). Lakes were characterized by differences in terms of anthropogenic pressure (i.e. urbanization, cultivated crops, livestock grazing) and potential “background noise” levels (i.e. nutrient concentration, pH, microbial concentration). Our aim was to assess nitrogen isotopic variations in fragments of Ulva lactuca specimens after 48 h of submersion to identify and locate the origins of nitrogen pollutants affecting each lake. δ¹⁵N were obtained for replicated specimens of U. lactuca spatially distributed to cover the entire surface of each lake, previously collected from a benchmark, unpolluted site. In order to reduce the environmental background noise on isotopic observations, a Bayesian hierarchical model relating isotopic variation to environmental covariates and random spatial effects was used to describe and understand the distribution of isotopic signals in each lake.Our procedure (i) allowed to remove background noise and confounding effects from the observed isotopic signals; (ii) allowed to detect “hidden” pollution sources that would not be detected when not accounting for the confounding effect of environmental background noise; (iii) produced maps of the three lakes providing a clear representation of the isotopic signal variation even where background noise was high. Maps were useful to locate nitrogen pollution sources, identify the origin of the dissolved nitrogen and quantify the extent of pollutants, showing localized organic pollution impacting Sabaudia and Fogliano, but not Caprolace. This method provided a clear characterization of both intra- and inter-lake anthropogenic pressure gradients, representing a powerful approach to the ecological indication and nitrogen pollution management in complex systems, as transitional waterbodies are.     

Experimental Evidence for a Hydride Transfer Mechanism in Plant Glycolate Oxidase Catalysis

In plants, glycolate oxidase is involved in the photorespiratory cycle, one of the major fluxes at the global scale. To clarify both the nature of the mechanism and possible differences in glycolate oxidase enzyme chemistry from C₃ and C₄ plant species, we analyzed kinetic parameters of purified recombinant C₃ (Arabidopsis thaliana) and C₄ (Zea mays) plant enzymes and compared isotope effects using natural and deuterated glycolate in either natural or deuterated solvent. The ¹²C/¹³C isotope effect was also investigated for each plant glycolate oxidase protein by measuring the ¹³C natural abundance in glycolate using natural or deuterated glycolate as a substrate. Our results suggest that several elemental steps were associated with an hydrogen/deuterium isotope effect and that glycolate α-deprotonation itself was only partially rate-limiting. Calculations of commitment factors from observed kinetic isotope effect values support a hydride transfer mechanism. No significant differences were seen between C₃ and C₄ enzymes.     

Novel strategies to overcome expression problems encountered with toxic proteins: Application to the production of Lac repressor proteins for NMR studies

NMR studies of structural aspects of allosteric regulation by the Lac repressor requires overexpression and isotope labeling of the protein. The size of the repressor makes it a challenging target, putting constraints on both expression conditions and sample preparation methods to overcome problems associated with studies of larger proteins by NMR. We optimized protocols for the production of deuterated functionally active thermostable dimeric Lac repressor and its core domain mutants. The Lac repressor core domain has never been obtained as a recombinant protein, possibly due to the observed toxicity to the host cells. We overcame the core domain induced toxicity by co-expression of this domain with the full length Lac repressor, combined with a stringent control of culture conditions. Significant overexpression was only obtained if during all stages of pre-culturing the bacteria were kept in their exponential growth phase at low density. The sensitivity of NMR measurements is dramatically affected by buffer conditions; we therefore used a thermofluor buffer optimization screen to determine the optimal buffer conditions. The combined thermofluor and NMR screening method yielded thermostable fully functional Lac repressor domain samples suitable for high-resolution NMR studies. The optimized procedures to adapt Escherichia coli to growth in D₂O, to overcome toxicity, and to optimize protein sample conditions provides a broad range of universally applicable techniques for production of larger proteins for NMR spectroscopy.     

盘县大洞古人类遗址的地层和古气候研究

贵州盘县大洞的堆积物,自上而下可以分为3层。大洞主体堆积物为角砾堆积,来源于洞顶灰岩的崩塌,另外有人类遗物以及洞外片流作用带入的泥砂。人类化石、旧石器和哺乳动物化石主要分布在第2层角砾堆积中。堆积物在洞厅各部位均有相应分布,属同一沉积序列。已知的早期人类活动时间大约从260ka．B．P．开始,到142ka．B．P．前后结束,对应于深海氧同位素的第6、7阶段。角砾层下段,发育一套遭强烈溶蚀和风化的堆积物,地球化学分析结果显示其湿热的沉积环境,时间大约在260—180ka B．P．期间,对应于深海氧同位素第7阶段;大洞堆积物在气候变化周期上与黄土、青藏高原冰期序列可以对比。     

Utilisation des isotopes stables pour l’identification de l’origine du carbone dans la datation du charbon de bois du Paléolithique

Recent years have brought many results of radiocarbon dating the earliest periods of the Upper Palaeolithic that can bring light on the origins of figurative art by Sapiens or Neanderthals. These dates are often close to the limit of the field of radiocarbon dating; because they require measurements of the lowest amounts of radiocarbon, controls are particularly essential. Here we examine the case of the dating of charcoal, whose identification after decontamination is difficult. We suggest a method that does not require additional manipulation to determine whether carbon comes exclusively from charcoal: using the proportion of stable carbon isotopes 13C/12C which is often regarded as a signature (δ13C).     

Salinities, not diets, affect strontium/calcium ratios in otoliths of Anguilla japonica

Although otolith Strontium (Sr)/calcium (Ca) ratios have been widely used to reconstruct the past salinity environmental history of anguillid eels, factors affecting the Sr/Ca ratios in otoliths are incompletely understood. Japanese Eel (Anguilla japonica) elvers (mean length 54.7 ± 2.1 mm) were collected in the estuary during their upstream migration and reared at 5 different salinities (0, 5, 15, 25, and 35 psu) and 3 types of feeding conditions (formulated feed, tubifex, and starvation) for 30 days to evaluate the effects of salinity and diets on otolith Sr/Ca ratios. Ca and Sr concentrations in the ambient water significantly increased with salinity (SAL) as [Ca] _water = 15.50SAL − 5.56, and [Sr] _water = 0.21SAL + 0.03, respectively. Sr/Ca ratios in otoliths increased with salinity (SAL) of the rearing water as [(Sr/Ca) × 1000] _otolith = 0.091SAL + 3.790. In diets, Sr/Ca ratios were 4 times higher in tubifex than in formulated feed. However, in otoliths, ANOVA indicated that Sr/Ca ratios did not differ significantly between groups fed on tubifex or formulated feed (p = 0.118). Otolith Sr/Ca ratios were negatively correlated with fish growth rates while the growth rates differed significantly among rearing conditions with different salinities and diets. Partition coefficients of the Sr/Ca ratios from ambient water to fish tissues and otoliths significantly increased with salinity. The Sr/Ca ratios of Japanese Eel otoliths thus were positively correlated with the ambient salinity and decreased with increasing fish growth rate, but was not affected by fish diet.     

Stable isotope pilot study of exchangeable copper kinetics in human blood plasma

To develop further our understanding of initial dietary copper metabolism, a method has been developed to separate plasma copper that is bound to albumin, from that bound to ceruloplasmin. This method has been tested using plasma samples from a pilot study involving six human volunteers who consumed 3mg oral doses of the stable isotope (65)Cu and gave blood samples at timed intervals up to 7 days. The results suggest that this method can be used to monitor dynamic fluctuations in newly absorbed copper over a short time frame.