Stable isotope analysis in soil prospection reveals the type of historic land-use under contemporary temperate forests in Europe.

The determination of δ13C and δ15N values is a common method in archaeological isotope analysis-in studying botanical and human remains, dietary practices, and less typically soils (to understand methods of agricultural cultivation, including fertilization). Stable isotope measurements are also commonly used in ecological studies to distinguish different ecosystems and to trace diachronic processes and biogeochemical mechanisms, however, the application of this method in geochemical prospection, for determining historic land-use impact, remains unexplored. The study at hand focuses on a deserted site of a Cistercian manor, dating from the thirteenth to fifteenth centuries. Isotopic measurements of anthropogenically influenced soils have been compared to approximately 400 archaeobotanical, soil, and sediment samples collected globally. The results reveal the potential of isotope measurements in soil to study the impact of past land use as isotope measurements identify specific types of agricultural activities, distinguishing crop production or grazing. δ13C and δ15N ratios also likely reflect fertilization practices and-in this case-the results indicate the presence of cereal cultivation (C3 cycle plants) and fertilization and that the site of the medieval manor was primarily used for grain production rather than animal husbandry.

Land cover and space use influence coyote carnivory: evidence from stable-isotope analysis.

For many species, the relationship between space use and diet composition is complex, with individuals adopting varying space use strategies such as territoriality to facilitate resource acquisition. Coyotes (Canis latrans) exhibit two disparate types of space use; defending mutually exclusive territories (residents) or moving nomadically across landscapes (transients). Resident coyotes have increased access to familiar food resources, thus improved foraging opportunities to compensate for the energetic costs of defending territories. Conversely, transients do not defend territories and are able to redirect energetic costs of territorial defense towards extensive movements in search of mates and breeding opportunities. These differences in space use attributed to different behavioral strategies likely influence foraging and ultimately diet composition, but these relationships have not been well studied. We investigated diet composition of resident and transient coyotes in the southeastern United States by pairing individual space use patterns with analysis of stable carbon (δ13C) and nitrogen (δ15N) isotope values to assess diet. During 2016-2017, we monitored 41 coyotes (26 residents, 15 transients) with GPS radio-collars along the Savannah River area in the southeastern United States. We observed a canopy effect on δ13C values and little anthropogenic food in coyote diets, suggesting 13C enrichment is likely more influenced by reduced canopy cover than consumption of human foods. We also observed other land cover effects, such as agricultural cover and road density, on δ15N values as well as reduced space used by coyotes, suggesting that cover types and localized, resident-like space use can influence the degree of carnivory in coyotes. Finally, diets and niche space did not differ between resident and transient coyotes despite differences observed in the proportional contribution of potential food sources to their diets. Although our stable isotope mixing models detected differences between the diets of resident and transient coyotes, both relied mostly on mammalian prey (52.8%, SD = 15.9 for residents, 42.0%, SD = 15.6 for transients). Resident coyotes consumed more game birds (21.3%, SD = 11.6 vs 13.7%, SD = 8.8) and less fruit (10.5%, SD = 6.9 vs 21.3%, SD = 10.7) and insects (7.2%, SD = 4.7 vs 14.3%, SD = 8.5) than did transients. Our findings indicate that coyote populations fall on a feeding continuum of omnivory to carnivory in which variability in feeding strategies is influenced by land cover characteristics and space use behaviors.

Leaching sources and mechanisms of different nitrogen species from bioretention systems.

At present, nitrogen (N) leaching from bioretention systems (BRSs) has become a key issue, imposing constraints on their application, a consequence of N dynamics of both inflow and legacy N at different time scales. In this study, the distinct sources (IL: immediate leaching, FL: fast leaching, SL: slow leaching) and the principal transformation processes of different N species (i.e., NH4+, NO3- and DON) leaching originating from inflow and legacy of BRSs were firstly unveiled by various 15N species labeling (i.e., 15N-NH4+, 15N-NO3- and 15N-DON). Results indicate that: NH4+ leaching was primarily caused by FL from influent organic N and SL from influent NH4+, with mineralization being the main transformation process influencing NH4+ leaching; NO3- leaching primarily originated from SL, with the major proportion attributed to the influent organic N in SL, autotrophic and heterotrophic nitrification were the main influencing factors; DON leaching primarily originated from SL, with similar proportions coming from influent organic N, NH4+, and NO3-, inorganic N assimilation was the principal transformation process affecting DON leaching. This study provides an effective framework for apportioning the leaching sources of different N species, providing valuable insights for the implementation of both inflow and legacy N leaching control measures.

Elevated Temperature Effects on Protein Turnover Dynamics in Arabidopsis thaliana Seedlings Revealed by 15N-Stable Isotope Labeling and ProteinTurnover Algorithm.

Global warming poses a threat to plant survival, impacting growth and agricultural yield. Protein turnover, a critical regulatory mechanism balancing protein synthesis and degradation, is crucial for the cellular response to environmental changes. We investigated the effects of elevated temperature on proteome dynamics in Arabidopsis thaliana seedlings using 15N-stable isotope labeling and ultra-performance liquid chromatography-high resolution mass spectrometry, coupled with the ProteinTurnover algorithm. Analyzing different cellular fractions from plants grown under 22 °C and 30 °C growth conditions, we found significant changes in the turnover rates of 571 proteins, with a median 1.4-fold increase, indicating accelerated protein dynamics under thermal stress. Notably, soluble root fraction proteins exhibited smaller turnover changes, suggesting tissue-specific adaptations. Significant turnover alterations occurred with redox signaling, stress response, protein folding, secondary metabolism, and photorespiration, indicating complex responses enhancing plant thermal resilience. Conversely, proteins involved in carbohydrate metabolism and mitochondrial ATP synthesis showed minimal changes, highlighting their stability. This analysis highlights the intricate balance between proteome stability and adaptability, advancing our understanding of plant responses to heat stress and supporting the development of improved thermotolerant crops.

Variation in Nitrogen Utilization and Nutrient Composition across Various Organs under Different Strip Logging Management Models in Moso Bamboo (Phyllostachys edulis) Forest.

The rapid restoration and renewal of the moso bamboo logging zone after strip logging has emerged as a key research area, particularly regarding whether nutrient accumulation and utilization in reserve zones can aid in the restoration and regeneration of the logging zone. In this study, a dynamic 15N isotope tracking experiment was conducted by injecting labeled urea fertilizer into bamboo culms. Logging zones and reserve zones of 6 m, 8 m, and 10 m widths were established. The conventional selective logging treatment served as a control (Con). Measurements were taken in May and October to assess the differences in nitrogen accumulation ability, utilization rates, and nutrient content across different organs in bamboo forests at different growth stages and under different treatments. Principal component analysis was conducted to evaluate and determine the importance of each indicator and strip logging treatment comprehensively. The results showed that various bamboo organs exhibited higher nitrogen accumulation and utilization rates during the peak growth period compared to the late growth period. Leaves had the highest nitrogen accumulation and utilization rates than the other organs. The average C content in various bamboo organs under different logging treatments exhibited subtle differences, irrespective of variation in logging width treatments. Bamboo culm exhibited the highest carbon accumulation. The C content in various bamboo organs was higher during the peak growth period than in the late growth period. The nitrogen content peaked in the leaves during the two growth stages and was significantly higher compared to the other organs. Most bamboo organs in the logging zones exhibited relatively higher nitrogen content than in the reserve zone and Con group. The P content was highest in bamboo leaves compared with other organs across the different strip logging treatments. Principal component analysis revealed relatively high absolute values of the coefficients for the C content, bamboo stump C content, and culm Ndff%. Log8 and Res10 zones had the highest comprehensive evaluation scores, indicating that Log8 and Res10 had the best effect on the promotion of nitrogen utilization and nutrient accumulation in various organs of moso bamboo.

Effects of straw management and N levels on gross nitrogen transformations in fluvo-aquic soil of the North China Plain.

Straw incorporation with nitrogen (N) fertilization is crucial for enhancing soil fertility and minimizing negative environmental impacts by altering the magnitude and direction of soil N transformation processes. However, the response of soil N transformations to long-term carbon (C) and N inputs, and their primary driving factors, remain poorly understood. Thus, a 15N tracing study was conducted to investigate the effects of straw incorporation (AS) and straw removal (NS) with N levels of 0, 150 and 250 kg N ha-1 per season (N0, N150 and N250) on gross N transformation rates in the North China Plain after 6-year trial. Results indicated that at N0, AS significantly increased soil microbial immobilization of nitrate (NO3--N, INO3) and autotrophic nitrification rates (ONH4) compared to NS. With N fertilization, AS increased gross N immobilization (Itotal), ammonium-N immobilization (NH4+-N, INH4), net NH4+-N immobilization (InetNH4) and net NH4+-N absorption rates (AnetNH4). Specifically, at N150, AS significantly increased recalcitrant organic N mineralization rate (MNrec), while significantly reducing ONH4, labile organic N mineralization (MNlab), and gross N mineralization rates (Mtotal). At N250, AnetNH4, MNlab, MNrec and ONH4 under AS were significantly higher than under NS. Nitrogen application significantly increased ONH4, Itotal and INO3 under two straw management practices, and enhanced INH4 and InetNH4 under AS. Compared to N250, N150 significantly increased INH4 and InetNH4 under AS, while decreasing Mtotal. Opposite results were observed under NS. Meanwhile, NO3--N and dissolved organic carbon (DOC) were master factors controlling immobilization, total nitrogen (TN), hydrolysable NH4+-N (HNN) and stable organic N significantly affected AnetNH4, while labile organic N were the key environmental factors affecting MNrec, all of which positively influenced the rates of assimilation, mineralization and clay mineral adsorption. Overall, this study provides new insights into reducing N fertilization under straw incorporation by quantifying soil N transformation processes.

The differential assimilation of nitrogen fertilizer compounds by soil microorganisms.

The differential soil microbial assimilation of common nitrogen (N) fertilizer compounds into the soil organic N pool is revealed using novel compound-specific amino acid (AA) 15N-stable isotope probing. The incorporation of fertilizer 15N into individual AAs reflected the known biochemistry of N assimilation-e.g. 15N-labelled ammonium (15NH4+) was assimilated most quickly and to the greatest extent into glutamate. A maximum of 12.9% of applied 15NH4+, or 11.7% of 'retained' 15NH4+ (remaining in the soil) was assimilated into the total hydrolysable AA pool in the Rowden Moor soil. Incorporation was lowest in the Rowden Moor 15N-labelled nitrate (15NO3-) treatment, at 1.7% of applied 15N or 1.6% of retained 15N. Incorporation in the 15NH4+ and 15NO3- treatments in the Winterbourne Abbas soil, and the 15N-urea treatment in both soils was between 4.4% and 6.5% of applied 15N or 5.2% and 6.4% of retained 15N. This represents a key step in greater comprehension of the microbially mediated transformations of fertilizer N to organic N and contributes to a more complete picture of soil N-cycling. The approach also mechanistically links theoretical/pure culture derived biochemical expectations and bulk level fertilizer immobilization studies, bridging these different scales of understanding.

Seasonal isotopic and isotopomeric signatures of nitrous oxide produced microbially in a eutrophic estuary.

Anthropogenic input of excess nutrients stimulates massive nitrous oxide (N2O) production in estuaries with distinct seasonal variations. Here, nitrogen isotopic and isotopomeric signatures were utilized to investigate the seasonal dynamics of N2O production and nitrification at the middle reach of the eutrophic Pearl River Estuary in the south of China. Elevated N2O production primarily via ammonia oxidation (> 1 nM-N d-1) occurred from April to November, along with increased temperature and decreased dissolved oxygen concentration. This consistently oxygenated water column showed active denitrification, contributing 20-40 % to N2O production. The water column microbial N2O production generally constituted a minor fraction (10-15 %) of the estuarine water-air interface efflux, suggesting that upstream transport and tidal dilution regulated the dissolved N2O inventory in the middle reach of the estuary. Nitrification (up to 3000 nM-N d-1) played a critical role in bioavailable nitrogen conversion and N2O production, albeit with N2O yields below 0.05 %.

Comparative study of urea-15N fate in pure bamboo and bamboo-broadleaf mixed forests.

Objectives: Bamboo is a globally significant plant with ecological, environmental, and economic bene-fits. Choosing suitable native tree species for mixed planting in bamboo forests is an effective measure for achieving both ecological and economic benefits of bamboo forests. However, little is currently known about the impact of bamboo forests on nitrogen cycling and utilization efficiency after mixing with other tree species. Therefore, our study aims to compare the nitrogen cycling in pure bamboo forests with that in mixed forests. Methods: Through field experiments, we investigated pure Qiongzhuea tumidinoda forests and Q. tumidinoda-Phellodendron chinense mixed forests, and utilized 15N tracing technology to explore the fertilization effects and fate of urea-15N in different forest stands. Results: The results demonstrated the following: 1) in both forest stands, bamboo culms account for the highest biomass percentage (42.99%-51.86%), while the leaves exhibited the highest nitrogen concentration and total nitrogen uptake (39.25%-44.52%/29.51%-33.21%, respectively) Additionally, the average nitrogen uptake rate of one-year-old bamboo is higher (0.25 mg kg-1 a-1) compared to other age groups. 2) the urea-15N absorption in mixed forests (1066.51-1141.61 g ha-1, including 949.65-1000.07 g ha-1 for bamboo and 116.86-141.54 g ha-1 for trees) was significantly higher than that in pure forests (663.93-727.62 g ha-1, P<0.05). Additionally, the 15N recovery efficiency of culms, branches, leaves, stumps, and stump roots in mixed forests was significantly higher than that in pure forests, with increases of 43.14%, 69.09%, 36.84%, 51.63%, 69.18%, 34.60%, and 26.89%, respectively. 3) the recovery efficiency of urea-15N in mixed forests (45.81%, comprising 40.43% for bamboo and 5.38% for trees) and the residual urea-15N recovery rate in the 0-60 cm soil layer (23.46%) are significantly higher compared to those in pure forests (28.61%/18.89%). This could be attributed to the nitrogen losses in mixed forests (30.73%, including losses from ammonia volatilization, runoff, leaching, and nitrification-denitrification) being significantly lower than those in pure forests (52.50%). Conclusion: These findings suggest that compared to pure bamboo forests, bamboo in mixed forests exhibits higher nitrogen recovery efficiency, particularly with one-year-old bamboo playing a crucial role.

Does long-term drought or repeated defoliation affect seasonal leaf N cycling in young beech trees?

Forest trees adopt effective strategies to optimize nitrogen (N) use through internal N recycling. In the context of more recurrent environmental stresses due to climate change, the question remains of whether increased frequency of drought or defoliation threatens this internal N recycling strategy. We submitted 8-year-old beech trees to 2 years of either severe drought (Dro) or manual defoliation (Def) to create a state of N starvation. At the end of the second year before leaf senescence, we labeled the foliage of the Dro and Def trees, as well as that of control (Co) trees, with 15N-urea. Leaf N resorption, winter tree N storage (total N, 15N, amino acids, soluble proteins) and N remobilization in spring were evaluated for the three treatments. Defoliation and drought did not significantly impact foliar N resorption or N concentrations in organs in winter. Total N amounts in Def tree remained close to those in Co tree, but winter N was stored more in the branches than in the trunk and roots. Total N amount in Dro trees was drastically reduced (-55%), especially at the trunk level, but soluble protein concentrations increased in the trunk and fine roots compared with Co trees. During spring, 15N was mobilized from the trunk, branches and twigs of both Co and Def trees to support leaf growth. It was only provided through twig 15N remobilization in the Dro trees, thus resulting in extremely reduced Dro leaf N amounts. Our results suggest that stress-induced changes occur in N metabolism but with varying severity depending on the constraints: within-tree 15N transport and storage strategy changed in response to defoliation, whereas a soil water deficit induced a drastic reduction of the N amounts in all the tree organs. Consequently, N dysfunction could be involved in drought-induced beech tree mortality under the future climate.

Hyperpolarization of 15N-Pyridinium by Using Parahydrogen Enables Access to Reactive Oxygen Sensors and Pilot In Vivo Studies.

Magnetic resonance with hyperpolarized contrast agents is one of the most powerful and noninvasive imaging platforms capable for investigating in vivo metabolism. While most of the utilized hyperpolarized agents are based on 13C nuclei, a milestone advance in this area is the emergence of 15N hyperpolarized contrast agents. Currently, the reported 15N hyperpolarized agents mainly utilize the dissolution dynamic nuclear polarization (d-DNP) protocol. The parahydrogen enhanced 15N probes have proven to be elusive and have been tested almost exclusively in organic solvents. Herein, we designed a reaction based reactive oxygen sensor 15N-boronobenzyl-2-styrylpyridinium (15N-BBSP) which can be hyperpolarized with para-hydrogen. Reactive oxygen species plays a vital role as one of the essential intracellular signalling molecules. Disturbance of the H2O2 level usually represents a hallmark of pathophysiological conditions. This H2O2 probe exhibited rapid responsiveness toward H2O2 and offered spectrally resolvable chemical shifts. We also provide strategies to bring the newly developed probe from the organic reaction solution into a biocompatible injection buffer and demonstrate the feasibility of in vivo 15N signal detection. The present work manifests its great potential not only for reaction based reactive sensing probes but also promises to serve as a platform to develop other contrast agents.

Synthesis of novel indol-3-acetamido analogues as potent anticancer agents, biological evaluation and molecular modeling studies.

Cannabinoids bind to cannabinoid receptors CB1 and CB2 and their antitumoral activity has been reported against some various cancer cell lines. Some synthetic cannabinoids possessing indole rings such as JWH-015 and JWH-133 particularly bind to the cannabinoid CB2 receptor and it was reported that they inhibit the proliferation and growth of various cancer cells without their psychoactive effects. However, the pharmacological action mechanisms of the cannabinoids are completely unknown. In this study, we report the synthesis of some new cannabinoidic novel indoles and evaluate their anticancer activity on various cancerous and normal cell lines (U87, RPMI 8226, HL60 and L929) using several cellular and molecular assays including MTT assay, real-time q-PCR, scratch assay, DAPI assay, Annexin V-PE/7AAD staining, caspase3/7 activity tests. Our findings indicated that compounds 7, 10, 13, 16, and 17 could reduce cell viability effectively. Compound 17 markedly increased proapoptotic genes (BAX, BAD, and BIM), tumor suppressor gene (p53) expression levels as well as the BAX/BCL-2 ratio in U87 cells. In addition, 17 inhibited cell migration. Based on these results, 17 was chosen for determining the mechanism of cell death in U87 cells. DAPI and Annexin V-7AAD staining results showed that 17 induced apoptosis, moreover activated caspase 3/7 significantly. Hence, compound 17, was selected as a lead compound for further pharmacomodulation. To rationalize the observed biological activities of 17, our study also included a comprehensive analysis using molecular docking and MD simulations. This integrative approach revealed that 17 fits tightly into the active site of the CB2 receptor and is involved in key interactions that may be responsible for its anti-proliferative effects.

Contamination of depressional wetlands in the Mpumalanga Lake District of South Africa near a global emission hotspot.

The Mpumalanga Lake District (MLD) of South Africa hosts a regionally unique cluster of water bodies of great importance for wetland biodiversity. It is also located close to a global hotspot for coal-fired power station emissions but the local impacts from these sources of pollution are poorly understood. Sediment cores from three contrasting wetlands were 210Pb dated and analysed for a range of contaminants linked to fossil fuel combustion, including trace elements, Hg, sulphur and spheroidal carbonaceous fly-ash particles (SCPs). At the two sites with pre-industrial (1900) baseline sediments, Pb, Zn and especially Cr concentrations and fluxes showed significant increases in the impact period (post-1975). Mercury showed the greatest proportional increase in flux (>4-fold) of all trace metals. Mercury and sulphur concentrations and fluxes showed highly significant correlations with emissions over the corresponding periods, while SCPs in sediments also closely tracked emissions. In a global context, levels of sediment contamination are relatively minor compared with other heavily industrialised regions, with only Cr exceeding the sediment Probable Effects Concentration for biological impact post-1975. Despite the relatively large increases in Hg, concentrations do not reach the Threshold Effects Concentration. The unexpectedly low levels of contamination may be due to i) low levels of many trace contaminants in South African coals compared to global averages, ii) prevailing recirculation patterns which transport pollution away from the study area during the wet season, minimising wet deposition, and iii) pollutant remobilisation through desiccation of wetlands or volatilization. The effects of hydrology and sediment accumulation rates lead to differential transport and preservation of organic-associated and more volatile contaminants (e.g. Hg, S) relative to non-volatile trace elements in wetlands of the MLD. The greatest fluxes of Hg and S are recorded in the site with the highest catchment: lake area ratio, lowest salinity and greatest sediment organic matter content.

15N-DNA stable isotope probing reveals niche differentiation of ammonia oxidizers in paddy soils.

Chemoautotrophic canonical ammonia oxidizers (ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB)) and complete ammonia oxidizers (comammox Nitrospira) are accountable for ammonia oxidation, which is a fundamental process of nitrification in terrestrial ecosystems. However, the relationship between autotrophic nitrification and the active nitrifying populations during 15N-urea incubation has not been totally clarified. The 15N-labeled DNA stable isotope probing (DNA-SIP) technique was utilized in order to study the response from the soil nitrification process and the active nitrifying populations, in both acidic and neutral paddy soils, to the application of urea. The presence of C2H2 almost completely inhibited NO3--N production, indicating that autotrophic ammonia oxidation was dominant in both paddy soils. 15N-DNA-SIP technology could effectively distinguish active nitrifying populations in both soils. The active ammonia oxidation groups in both soils were significantly different, AOA (NS (Nitrososphaerales)-Alpha, NS-Gamma, NS-Beta, NS-Delta, NS-Zeta and NT (Ca. Nitrosotaleales)-Alpha), and AOB (Nitrosospira) were functionally active in the acidic paddy soil, whereas comammox Nitrospira clade A and Nitrosospira AOB were functionally active in the neutral paddy soil. This study highlights the effective discriminative effect of 15N-DNA-SIP and niche differentiation of nitrifying populations in these paddy soils. KEY POINTS: • 15N-DNA-SIP technology could effectively distinguish active ammonia oxidizers. • Comammox Nitrospira clade A plays a lesser role than canonical ammonia oxidizers. • The active groups in the acidic and neutral paddy soils were significantly different.

Nitrogen uptake rates and phytoplankton composition across contrasting North Atlantic Ocean coastal regimes north and south of Cape Hatteras.

Understanding nitrogen (N) uptake rates respect to nutrient availability and the biogeography of phytoplankton communities is crucial for untangling the complexities of marine ecosystems and the physical, biological, and chemical forces shaping them. In the summer of 2016, we conducted measurements of bulk microbial uptake rates for six 15N-labeled substrates: nitrate, nitrite, ammonium, urea, cyanate, and dissolve free amino acids across distinct marine provinces, including the continental shelf of the Mid-and South Atlantic Bights (MAB and SAB), the Slope Sea, and the Gulf Stream, marking the first instance of simultaneously measuring six different N uptake rates in this dynamic region. Total measured N uptake rates were lowest in the Gulf Stream followed by the SAB. Notably, the MAB exhibited significantly higher N uptake rates compared to the SAB, likely due to the excess levels of pre-existing phosphorus present in the MAB. Together, urea and nitrate uptake contributed approximately 50% of the total N uptake across the study region. Although cyanate uptake rates were consistently low, they accounted for up to 11% of the total measured N uptake at some Gulf Stream stations. Phytoplankton groups were identified based on specific pigment markers, revealing a dominance of diatoms in the shelf community, while Synechococcus, Prochlorococcus, and pico-eukaryotes dominated in oligotrophic Gulf Stream waters. The reported uptake rates in this study were mostly in agreement with previous studies conducted in coastal waters of the North Atlantic Ocean. This study suggests there are distinct regional patterns of N uptake in this physically dynamic region, correlating with nutrient availability and phytoplankton community composition. These findings contribute valuable insights into the intricate interplay of biological and chemical factors shaping N dynamics in disparate marine ecosystems.

N-(Sulfonio)Sulfilimine Reagents: Non-Oxidizing Sources of Electrophilic Nitrogen Atom for Skeletal Editing.

The one-pot synthesis of λ4-dibenzothiophen-5-imino-N-dibenzothiophenium triflate (1) in multigram scale is reported. This compound reacts with Rh2(esp)2 (esp=α,α,α',α'-tetramethyl-1,3-benzenedipropionic acid) generating a Rh-coordinated sulfonitrene species, which is able to transfer the electrophilic nitrene moiety to olefins. When indenes are used as substrates, isoquinolines are obtained in good yields. We assumed that after formation of the corresponding N-sulfonio aziridine, a ring expansion occurs via selective C-C bond cleavage and concomitant elimination of dibenzothiophene. Unexpectedly, a similar protocol transforms 1-arylcyclobutenes into 1-cyano-1-arylcyclopropanes. Our calculations indicate that aziridination is not favored in this case; instead, sulfilimine-substituted cyclobutyl carbocations are initially formed, and these evolve to the isolated cyclopropanes via ring contraction. Both procedures are operationally simple, tolerate a range of functional groups, including oxidation-sensitive alcohols and aldehydes, and enable the convenient preparation of valuable 15N-labelled products. These results demonstrate the potential of 1 to provide alternative pathways for the selective transfer of N-atoms in organic molecules.

A study on the simultaneous determination of nitrogen content and 15N isotope abundance in plants using peak height intensities at m/z 28 and 29.

A method for simultaneous determination of nitrogen content and 15N isotope abundance in plants was established by Elemental analysis-gas isotope ratio mass spectrometry. Taking poplar leaves and l-glutamic acid as standards, nitrogen content was determined using the standard curve established by weighted least squares regression between the mass of nitrogen element and the total peak height intensity at m/z 28 and 29. Then the 15N isotope abundance was calculated with the peak height intensity at m/z 28 and 29. Through the comparison of several sets of experiments, the impact of mass discrimination effect, tin capsule consumables, isotope memory effect, and the quality of nitrogen on the results were assessed. The results showed that with a weight of 1/x2, the standard curve has a coefficient of determination (R2) of 0.9996. Compared to the traditional Kjeldahl method, the measured nitrogen content deviated less than 0.2 %, and the standard deviation (SD) was less than 0.2 %. Compared to the sodium hypobromite method, the 15N isotopic abundances differed less than 0.2 atom%15N, and the SD was less than 0.2 atom% 15N. The established method offers the advantages of being fast, simple, accurate, and high throughput, providing a novel approach for the simultaneous determination of nitrogen content and 15N isotope abundance in plant samples.

The impact of algal blooms on promoting in-situ N2O emissions: A case in Zhanjiang bay, China.

Under the influence of many factors, such as climate change, anthropogenic eutrophication, and the development of aquaculture, the area and frequency of algal blooms have showed an increasing trend worldwide, which has become a challenging issue at present. However, the coupled relationship between nitrous oxide (N2O) and algal blooms and the underlying mechanisms remain unclear. To address this issue, 15N isotope cultures and quantitative polymerase chain reaction (qPCR) experiments were conducted in Zhanjiang Bay during algal and non-algal bloom periods. The results showed that denitrification and nitrification-denitrification were the two processes responsible for the in-situ production of N2O during algal and non-algal bloom periods. Stable isotope rate cultivation experiments indicated that denitrification and nitrification-denitrification were promoted in the water during the algal bloom period. The in-situ production of N2O during the algal bloom period was three-fold that during the non-algal bloom period. This may be because fresh particulate organic matter (POM) from the organisms responsible for the algal bloom provides the necessary anaerobic and hypoxic environment for denitrification and nitrification-denitrification in the degradation environment. Additionally, a positive linear correlation between N2O concentrations and ammonia-oxidizing bacteria (AOB) and denitrifying bacteria (nirK and nirS) also supported the significant denitrification and nitrification-denitrification occurring in the water during the algal bloom period. However, the algal bloom changed the main process for the in-situ production of N2O, wherein it shifted from denitrification during the non-algal bloom period to nitrification-denitrification during the algal bloom period. The results of our study will improve our understanding of the processes responsible for the in-situ production of N2O during the algal bloom period, and can help formulate effective policies to mitigate N2O emissions in the bay.

Production of Isotopically Labeled KRAS4b.

Isotopically labelled proteins are important reagents in structural biology as well as in targeted drug development. The field continues to advance with complex multi-isotope labeling. We have combined our experience in high-level soluble KRAS4b expression with protocols for isotope incorporation, to achieve reliable and efficient approaches for several labeling strategies. Typical experiments achieve nearly 100% 15N incorporation, with yields in the range of 1.3-24.6 mg/L (median = 6.4 mg/L, n = 53). Improvements in the growth parameters in the presence of deuterium reduce the standard time of fermentation from 5 days to 3 days by modifying the medium used during the weaning process. The methods described are compatible with multi-isotope labeling and site-specific labeling.

Seeing further into the early steps of the endangered atlantic goliath grouper (Epinephelus itajara): Eye lenses high resolution isotopic profiles reveal ontogenetic trophic and habitat shifts.

Estuarine mangroves are often considered nurseries for the Atlantic Goliath grouper juveniles. Yet, the contributions of different estuarine primary producers and habitats as sources of organic matter during early ontogenetic development remain unclear. Given the species' critically endangered status and protection in Brazil, obtaining biological samples from recently settled recruits in estuaries is challenging. In this study, we leveraged a local partnership with fishers and used stable isotope (C and N) profiles from the eye lenses of stranded individuals or incidentally caught by fishery to reconstruct the trophic and habitat changes of small juveniles. The eye lens grows by the apposition of protein-rich layers. Once these layers are formed, they become inert, allowing to make inferences on the trophic ecology and habitat use along the development of the individual until its capture. We used correlations between fish size and the entire eye lens size, along with estuarine baselines, to reconstruct the fish size and trophic positions for each of the lens layers obtained. We then used dominant primary producers and basal sources from mangrove sheltered, exposed estuarine and marine habitats to construct an ontogenetic model of trophic and habitat support changes since maternal origins. Our model revealed marine support before the juveniles reached 25 mm (standard length), followed by a rapid increase in reliance on mangrove sheltered sources, coinciding with the expected size at settlement. After reaching 60 mm, individuals began to show variability. Some remained primarily supported by the mangrove sheltered area, while others shifted to rely more on the exposed estuarine area around 150 mm. Our findings indicate that while mangroves are critical for settlement, as Goliath grouper juveniles grow, they can utilize organic matter produced throughout the estuary. This underscores the need for conservation strategies that focus on seascape connectivity, as protecting just one discrete habitat may not be sufficient to preserve this endangered species and safeguard its ecosystem functions.