Unexpected 13N concentrations in ISIS synchrotron room air.

The specific activity of air in the large open room housing the 800-MeV proton synchrotron of the ISIS Spallation Neutron and Muon Source has been measured. Air from several positions within the ISIS synchrotron room was sucked through a long flexible tube, and run past a shielded HPGe gamma-ray detector outside the synchrotron room. In spite of an expectation that 13N should be the largest component of the overall activity in the air, the results of the measurements are consistent with the presence in the air of 11C and 41Ar only, and suggest that the activity in the air is mostly created not in the synchrotron room itself but in the massive shielding monoliths around the neutron-producing targets, monoliths through which ventilation air is drawn into the synchrotron room. Typical specific activities of 11C and 41Ar in the air in the synchrotron room are ∼0.10 and ∼0.03 Bq cm-3 respectively, the upper limit for 13N being at most ∼0.01 Bq cm-3.

Leaf 13C and 15N composition shedding light on easing drought stress through partial K substitution by Na in eucalyptus species.

This work aimed to investigate the partial K-replacement by Na supply to alleviate drought-induced stress in Eucalyptus species. Plant growth, leaf gas exchange parameters, water relations, oxidative stress (H2O2 and MDA content), chlorophyll concentration, carbon (C) and nitrogen (N) isotopic leaf composition (δ13C and δ15N) were analyzed. Drought tolerant E. urophylla and E. camaldulensis showed positive responses to the partial K substitution by Na, with similar dry mass yields, stomatal density and total stomatal pore area relative to the well K-supplied plants under both water conditions, suggesting that 50% of the K requirements is pressing for physiological functions that is poorly substituted by Na. Furthermore, E. urophylla and E. camaldulensis up-regulated leaf gas exchanges, leading to enhanced long-term water use efficiency (WUEL). Moreover, the partial K substitution by Na had no effects on plants H2O2, MDA, δ13C and δ15N, confirming that Na, to a certain extent, can effectively replace K in plants metabolism. Otherwise, the drought-sensitive E. saligna species was negatively affected by partial K replacement by Na, decreasing plants dry mass, even with up-regulated leaf gas exchange parameters. The exclusive Na-supplied plants showed K-deficient symptoms and lower growth, WUEL, and δ13C, besides higher Na accumulation, δ15N, H2O2 and MDA content.

N-glycan fingerprint predicts alpha-fetoprotein negative hepatocellular carcinoma: A large-scale multicenter study.

Alpha-fetoprotein (AFP)-negative hepatocellular carcinoma (ANHCC) patients account for more than 30% of the whole entity of HCC patients and are easily misdiagnosed. This three-phase study was designed to find and validate new ANHCC N-glycan markers which identified from The Cancer Genome Atlas (TCGA) database and noninvasive detection. Differentially expressed genes (DEGs) of N-glycan biosynthesis and degradation related genes were screened from TCGA database. Serum N-glycan structure abundances were analyzed using N-glycan fingerprint (NGFP) technology. Totally 1340 participants including ANHCC, chronic liver diseases and healthy controls were enrolled after propensity score matching (PSM). The Lasso algorithm was used to select the most significant N-glycan structures abundances. Three machine learning models [random forest (RF), support vector machine (SVM) and logistic regression (LR)] were used to construct the diagnostic algorithms. All 13N-glycan structure abundances analyzed by NGFP demonstrated significant and was enrolled by Lasso. Among the three machine learning models, LR algorithm demonstrated the best diagnostic performance for identifying ANHCC in training cohort (LR: AUC: 0.842, 95%CI: 0.784-0.899; RF: AUC: 0.825, 95%CI: 0.766-0.885; SVM: AUC: 0.610, 95%CI: 0.527-0.684). This LR algorithm achieved a high diagnostic performance again in the independent validation (AUC: 0.860, 95%CI: 0.824-0.897). Furthermore, the LR algorithm could stratify ANHCC into two distinct subgroups with high or low risks of overall survival and recurrence in follow-up validation. In conclusion, the biomarker panel consisting of 13N-glycan structures abundances using the best-performing algorithm (LR) was defined and indicative as an effective tool for HCC prediction and prognosis estimate in AFP negative subjects.

Fueling of a marine-terrestrial ecosystem by a major seabird colony.

Seabirds redistribute nutrients between different ecosystem compartments and over vast geographical areas. This nutrient transfer may impact both local ecosystems on seabird breeding islands and regional biogeochemical cycling, but these processes are seldom considered in local conservation plans or biogeochemical models. The island of Stora Karlsö in the Baltic Sea hosts the largest concentration of piscivorous seabirds in the region, and also hosts a large colony of insectivorous House martins Delichon urbicum adjacent to the breeding seabirds. We show that a previously reported unusually high insectivore abundance was explained by large amounts of chironomids-highly enriched in δ15N-that feed on seabird residues as larvae along rocky shores to eventually emerge as flying adults. Benthic ammonium and phosphate fluxes were up to 163% and 153% higher close to the colony (1,300 m distance) than further away (2,700 m) and the estimated nutrient release from the seabirds at were in the same order of magnitude as the loads from the largest waste-water treatment plants in the region. The trophic cascade impacting insectivorous passerines and the substantial redistribution of nutrients suggest that seabird nutrient transfer should be increasingly considered in local conservation plans and regional nutrient cycling models.

Interpreting ancient food practices: stable isotope and molecular analyses of visible and absorbed residues from a year-long cooking experiment.

Chemical analyses of carbonized and absorbed organic residues from archaeological ceramic cooking vessels can provide a unique window into the culinary cultures of ancient people, resource use, and environmental effects by identifying ingredients used in ancient meals. However, it remains uncertain whether recovered organic residues represent only the final foodstuffs prepared or are the accumulation of various cooking events within the same vessel. To assess this, we cooked seven mixtures of C3 and C4 foodstuffs in unglazed pots once per week for one year, then changed recipes between pots for the final cooking events. We conducted bulk stable-isotope analysis and lipid residue analysis on the charred food macro-remains, carbonized thin layer organic patina residues and absorbed lipids over the course of the experiment. Our results indicate that: (1) the composition of charred macro-remains represent the final foodstuffs cooked within vessels, (2) thin-layer patina residues represent a mixture of previous cooking events with bias towards the final product(s) cooked in the pot, and (3) absorbed lipid residues are developed over a number of cooking events and are replaced slowly over time, with little evidence of the final recipe ingredients.

Correlation of fluorescence microscopy, electron microscopy, and NanoSIMS stable isotope imaging on a single tissue section.

Correlative light and electron microscopy allows localization of specific molecules at the ultrastructural level in biological tissue but does not provide information about metabolic turnover or the distribution of labile molecules, such as micronutrients. We present a method to directly correlate (immuno)fluorescent microscopy, (immuno)TEM imaging and NanoSIMS isotopic mapping of the same tissue section, with nanometer-scale spatial precision. The process involves chemical fixation of the tissue, cryo sectioning, thawing, and air-drying under a thin film of polyvinyl alcohol. It permits to effectively retain labile compounds and strongly increases NanoSIMS sensitivity for 13C-enrichment. The method is illustrated here with correlated distribution maps of a carbonic anhydrase enzyme isotype, ß-tubulin proteins, and 13C- and 15N-labeled labile micronutrients (and their anabolic derivates) within the tissue of a reef-building symbiotic coral. This broadly applicable workflow expands the wealth of information that can be obtained from multi-modal, sub-cellular observation of biological tissue.

Stable isotope analysis of multiple tissues from Hawaiian honeycreepers indicates elevational movement.

We have limited knowledge of the patterns, causes, and prevalence of elevational migration despite observations of seasonal movements of animals along elevational gradients in montane systems worldwide. While a third of extant Hawaiian landbird species are estimated to be elevational migrants this assumption is based primarily on early naturalist's observations with limited empirical evidence. In this study, we compared stable hydrogen isotopes (δ2H) of metabolically inert (feathers) and active (blood plasma, red blood cells) tissues collected from the same individual to determine if present day populations of Hawaiian honeycreepers undergo elevational movements to track areas of seasonally high flower bloom that constitute significant food resources. We also measured stable carbon isotopes (δ13C) and stable nitrogen isotopes (δ15N) to examine potential changes in diet between time periods. We found that the majority of 'apapane (Himatione sanguinea) and Hawai'i 'amakihi (Chlorodrepanis virens) captured at high elevation, high bloom flowering sites in the fall were not year-round residents at the capture locations, but had molted their feathers at lower elevations presumably in the summer after breeding. δ2H values of feathers for all individuals sampled were higher than blood plasma isotope values after accounting for differences in tissue-specific discrimination. We did not find a difference in the propensity of elevational movement between 'apapane and Hawai'i 'amakihi, even though the 'amakihi is considered more sedentary. However, consistent with a more generalist diet, δ15N values indicated that Hawai'i 'amakihi had a more diverse diet across trophic levels than 'apapane, and a greater reliance on nectar in the fall. We demonstrate that collecting multiple tissue samples, which grow at different rates or time periods, from a single individual can provide insights into elevational movements of Hawaiian honeycreepers over an extended time period.

Caste-specific nutritional differences define carbon and nitrogen fluxes within symbiotic food webs in African termite mounds.

Fungus-growing termites of the genus Macrotermes cultivate symbiotic fungi (Termitomyces) in their underground nest chambers to degrade plant matter collected from the environment. Although the general mechanism of food processing is relatively well-known, it has remained unclear whether the termites get their nutrition primarily from the fungal mycelium or from plant tissues partly decomposed by the fungus. To elucidate the flows of carbon and nitrogen in the complicated food-chains within the nests of fungus-growing termites, we determined the stable isotope signatures of different materials sampled from four Macrotermes colonies in southern Kenya. Stable isotopes of carbon revealed that the termite queen and the young larvae are largely sustained by the fungal mycelium. Conversely, all adult workers and soldiers seem to feed predominantly on plant and/or fungus comb material, demonstrating that the fungal symbiont plays a different nutritional role for different termite castes. Nitrogen stable isotopes indicated additional differences between castes and revealed intriguing patterns in colony nitrogen cycling. Nitrogen is effectively recycled within the colonies, but also a presently unspecified nitrogen source, most likely symbiotic nitrogen-fixing bacteria, seems to contribute to nitrogen supply. Our results indicate that the gut microbiota of the termite queen might be largely responsible for the proposed nitrogen fixation.

Variation of carbon and isotope natural abundances (δ15N and δ13C) of whole-plant sweet potato (Ipomoea batatas L.) subjected to prolonged water stress.

Sweet potato (Ipomoea batatas L.) is an important crop in the world, cultivated in temperate climates under low inputs. Drought changes the plant biomass allocation, together with the carbon and nitrogen isotopic composition (δ13C and δ15N), whose changes are faintly known in sweet potato crops. Here, we show the biomass allocation of eight sweet potato accessions submitted to drought during 3 months, using the δ13C, δ15N, carbon isotope discrimination (Δ13C), total carbon (TC) and water use efficiency (WUE) traits. The tolerant accessions had improved WUE, with higher TPB and TC. Storage roots and shoots had a heavier δ13C content under drought stress, with greater 13C fixation in roots. The Δ13C did not show a significant association with WUE. The δ15N values indicated a generalised N reallocation between whole-plant organs under drought, as a physiological integrator of response to environmental stress. This information can aid the selection of traits to be used in sweet potato breeding programs, to adapt this crop to climate change.

Auditory ossicles: a potential biomarker for maternal and infant health in utero.

Background: Carbon (δ13C) and nitrogen (δ15N) isotope ratios of collagen from teeth and bone are used to study human nutrition and health. As bones are constantly remodelling throughout life, isotopic values of bone collagen represent an average of several years. In contrast, human teeth do not remodel and their primary dentine contains only the isotopic data from the time of formation. In contrast to all other bones, human auditory ossicles also appear not to remodel. As they develop in utero and finish formation in the first 2 years of life, their collagen should also represent isotopic values of these two relatively short periods.Aim: By comparing δ13C and δ15N data from ossicles and incremental dentine, this study aims to investigate how two developmental periods of the ossicles, in utero and the first 2 years of life, reflect in collagen obtained from the ossicles.Subject and methods: Ossicle and tooth samples of 12 individuals aged 0.5 ± 0.4 years to 13 ± 1 years from the nineteenth century St. Peter's burial ground in Blackburn were collected and processed to obtain bulk bone and incremental dentine collagen which was measured for δ13C and δ15N.Results: Averaged δ13C and δ15N of ossicles are lower when compared to every age group except after 3 years of age. Average offset between ossicles and dentine of different groups ranges from 0.4-0.9‰ for δ13C and from 0.3-0.9‰ for δ15N, with highest counterbalance at birth and after the first 5 months after birth.Conclusions: There appears to be a systematic offset between the dentine and ossicle data. It seems that the second phase of development does not influence the isotopic values of collagen significantly and the data we are obtaining from ossicles represents the in utero period.

Stable isotopes reveal patterns of diet and mobility in the last Neandertals and first modern humans in Europe.

Correlating cultural, technological and ecological aspects of both Upper Pleistocene modern humans (UPMHs) and Neandertals provides a useful approach for achieving robust predictions about what makes us human. Here we present ecological information for a period of special relevance in human evolution, the time of replacement of Neandertals by modern humans during the Late Pleistocene in Europe. Using the stable isotopic approach, we shed light on aspects of diet and mobility of the late Neandertals and UPMHs from the cave sites of the Troisième caverne of Goyet and Spy in Belgium. We demonstrate that their diet was essentially similar, relying on the same terrestrial herbivores, whereas mobility strategies indicate considerable differences between Neandertal groups, as well as in comparison to UPMHs. Our results indicate that UPMHs exploited their environment to a greater extent than Neandertals and support the hypothesis that UPMHs had a substantial impact not only on the population dynamics of large mammals but also on the whole structure of the ecosystem since their initial arrival in Europe.

Nitrogen (protein) determination in single seeds by nuclear reactions.

In this work, a brief review on nuclear reaction methods aimed at nitrogen assessment in organic matter is given, followed by a detailed application using a deuteron beam and (d,p) and (d,α) induced reactions on 14N to determine nitrogen content in single seeds This study covers the analysis of five wheat and four maize stable varieties, which showed up good (linear) correlation coefficients in comparison with corresponding Kjeldahl nitrogen analysis for each species. This method of analysis turns out to be a fast and non-destructive technique for protein determination in cereals, besides its extendibility to other plant products for human nutrition.

Quantifying N-loss by root abscission: consequences for wheat N budgets and δ15N values.

Lower plant δ15N values relative to source δ15N are commonly attributed to 15N efflux. We determined the extent to which root abscission contributes to plant N-loss and consequences for plant δ15N. Wheat (Triticum aestivum L. cv. SST015) was grown in hydroponics with direct aeration, aeration constrained within a pipe and circulation of nutrient solution through sand, representing three levels of stability for root growth. The δ15N of nutrient solutions and root fragments were periodically determined, as well as root and shoot δ15N. Plants in solution had significantly more negative δ15N (-8.9 and -9.2‰) than plants in sand (-6.9‰), suggesting greater 15N-loss; root fragments were major biomass- (six-fold greater than root dry weight) and N-loss (two-fold greater than plant net N uptake) pathways in solution. These plants had more ephemeral roots and two-fold more root tips than the sand treatment. We estimated that root fragment loss decreased plant δ15N by at least -3.7, -2.6 and -1.0‰ in the direct, pipe and sand treatments, respectively. Positive nutrient solution δ15N in all treatments relative to the source δ15N suggests that plant N, probably derived from efflux, was present in solution. Despite this, root abscission and root turnover are also important N-loss pathways in plants, while plant δ15N values are probably influenced by a combination of root abscission and N efflux.

Determining δ15N-NO3- values in soil, water, and air samples by chemical methods.

Soil, water, and air NO3- pollution is a major environmental problem worldwide. Stable isotope analysis can assess the origin of NOx because different NOx sources carry different isotope signatures. Hence, using appropriate chemical methods to determine the δ15N-NOx values in different samples is important to improve our understanding of the N-NOx pollution and take possible strategies to manage it. Two modified chemical methods, the cadmium-sodium azide method and the VCl3-sodium azide method, were used to establish a comprehensive inventory of δ15N-NOx values associated with major NOx fluxes by the conversion of NO3- into N2O. Precision and limit of detection values demonstrated the robustness of these quantitative techniques for measuring δ15N-NOx. The standard deviations of the δ15N-NO3- values were 0.35 and 0.34‰ for the cadmium-sodium azide and VCl3-sodium azide methods. The mean δ15N-NO3- values of river water, soil extracts, and summer rain were 8.9 ± 3.3, 3.5 ± 3.5, and 3.3 ± 2.1‰, respectively. The δ15N-NO3- values of low concentration samples collected from coal-fired power plants, motor vehicles, and gaseous HNO3 was 20.3 ± 4.3, 5.6 ± 2.78, and 5.7 ± 3.6‰, respectively. There was a good correlation between the δ15N-NO3- compositions of standards and samples, which demonstrates that these chemical reactions can be used successfully to assess δ15N-NO3- values in the environment.

Earliest isotopic evidence in the Maya region for animal management and long-distance trade at the site of Ceibal, Guatemala.

This study uses a multiisotope (carbon, nitrogen, oxygen, and strontium) approach to examine early animal management in the Maya region. An analysis of faunal specimens across almost 2,000 years (1000 BC to AD 950) at the site of Ceibal, Guatemala, reveals the earliest evidence for live-traded dogs and possible captive-reared taxa in the Americas. These animals may have been procured for ceremonial functions based on their location in the monumental site core, suggesting that animal management and trade began in the Maya area to promote special events, activities that were critical in the development of state society. Isotopic evidence for animal captivity at Ceibal reveals that animal management played a greater role in Maya communities than previously believed.

Decreased ¹³N-labeled ammonia uptake in the ipsilateral and contralateral hemispheres following carotid endarterectomy.

Carotid artery plaques are a leading cause of ischemic stroke, and carotid endarterectomy (CEA) is one of the major treatment approaches for this disease. Changes in cerebral metabolism following CEA remain unclear. The present study aimed to evaluate the effect of cerebral ammonia metabolism following CEA using 13N­labeled ammonia positron emission tomography (PET) in humans. A total of 20 patients were enrolled in the present study, with a mean age of 59.5 years, comprising 16 males and four females. Of these patients, eight underwent right CEA and 12 underwent left CEA. The rate of carotid artery stenosis was between 50­69% in six of the patients, between 70­99% in 11 of the patients and was at 100% (thrombosis) in three of the patients, measured by computerised tomography digital subtraction angiography prior to CEA. 13N­labeled ammonia (137 MBq) PET scanning was performed prior and subsequent to CEA surgery for each patient. The first ammonia PET scan was performed 1 day prior to CEA, while the second PET scan was performed 1­4 weeks following CEA. Following injection of 13N­labeled ammonia, static PET was acquired for 10 min. The region of interest (ROI), covering the major cerebral hemisphere, was selected and ammonia uptake in the ROI was determined in the ipsilateral and contralateral hemispheres. No hyperperfusion syndrome was observed in the patients subsequent to CEA. No significant change in cerebral hemisphere ammonia uptake was observed between the ipsilateral and contralateral hemispheres prior to (ratio =0.98; P>0.01) or following (ratio =1.09; P>0.01) CEA. Ammonia uptake in the ipsilateral and contralateral hemispheres was significantly reduced to 23.2 and 23.5%, respectively, following CEA. Using 13N­labeled ammonia PET to evaluate cerebral ammonia metabolism following CEA in patients with severe carotid artery stenosis, the present study demonstrated that uptake of ammonia in the ipsilateral and contralateral hemispheres was significantly reduced.

Neutron spectra due (13)N production in a PET cyclotron.

Monte Carlo and experimental methods have been used to characterize the neutron radiation field around PET (Positron Emission Tomography) cyclotrons. In this work, the Monte Carlo code MCNPX was used to estimate the neutron spectra, the neutron fluence rates and the ambient dose equivalent (H*(10)) in seven locations around a PET cyclotron during (13)N production. In order to validate these calculations, H*(10) was measured in three sites and were compared with the calculated doses. All the spectra have two peaks, one above 0.1MeV due to the evaporation neutrons and another in the thermal region due to the room-return effects. Despite the relatively large difference between the measured and calculated H*(10) for one point, the agreement was considered good, compared with that obtained for (18)F production in a previous work.

Measuring fluxes of mineral nutrients and toxicants in plants with radioactive tracers.

Unidirectional influx and efflux of nutrients and toxicants, and their resultant net fluxes, are central to the nutrition and toxicology of plants. Radioisotope tracing is a major technique used to measure such fluxes, both within plants, and between plants and their environments. Flux data obtained with radiotracer protocols can help elucidate the capacity, mechanism, regulation, and energetics of transport systems for specific mineral nutrients or toxicants, and can provide insight into compartmentation and turnover rates of subcellular mineral and metabolite pools. Here, we describe two major radioisotope protocols used in plant biology: direct influx (DI) and compartmental analysis by tracer efflux (CATE). We focus on flux measurement of potassium (K(+)) as a nutrient, and ammonia/ammonium (NH3/NH4(+)) as a toxicant, in intact seedlings of the model species barley (Hordeum vulgare L.). These protocols can be readily adapted to other experimental systems (e.g., different species, excised plant material, and other nutrients/toxicants). Advantages and limitations of these protocols are discussed.

Quantification of nitrate sources to an urban stream using dual nitrate isotopes.

Human-engineered landscapes and subsequent altered hydrology affect the fate and transport of reactive nitrogen, particularly in urban watersheds. In this study, we used dual-nitrate isotopes and mixing model analysis (δ(15)N and δ(18)O of NO3(-)) to quantify nitrogen inputs from two sources concentrated in urban systems, sewage and atmospheric deposition. Analysis was conducted on samples collected from Nine Mile Run (Pittsburgh, PA) including over two years of samples collected biweekly and samples collected through the hydrographs of four storm events. Mixing models incorporated uncertainties in the isotopic composition of potential nitrate sources and resolved the relative proportions of nitrate inputs from each source using Bayesian techniques. The results indicate that up to 94% of nitrate in streamwater originated from sewage sources during baseflow conditions. During storms, atmospheric deposition was a substantial nitrate source (∼ 34%) to total event-based nitrate loads, although sewage-derived nitrate remained the dominant source (66%). The potential influence of denitrification was considered by incorporating associated isotopic fractionations into mixing models; up to 19% of sewage-derived samples showed the isotopic effects of denitrification. This study quantitatively delineates proportions of nitrate from different sources to urban streamwater, while incorporating remaining uncertainties in source endmember compositions.

Raman activated cell ejection for isolation of single cells.

We have optimized a Raman microscope to obtain a single cell Raman spectrum (SCRS) with 0.1 s acquisition time. SCRS with such short acquisition time has sufficient discriminatory ability and spectral reproducibility to differentiate cells incorporated with (13)C and (15)N and to classify five different types of bacteria isolated from the oral cavity. We also developed Raman activated cell ejection (RACE) that is assisted by laser induced forward transfer (LIFT). We have shown, for the first time, that the single cells of interest can be identified and then accurately isolated from complex microbial communities based on their SCRS. This approach can be used to sort single cells of target traits from complex samples (e.g., biofilms, soils, sludge, tissues).