Integrated Stopped-Flow Device for the Study of Porous Materials Using Hyperpolarized 129Xe NMR.

A simple, low-cost, and efficient device is proposed for the study of porous materials via NMR using small gas probes. Mainly built through additive manufacturing and being equipped with a radiofrequency solenoid microcoil, it only requires tiny quantities of sample and/or gas and is particularly suited for hyperpolarized xenon. The performances of this device have been accessed on a commercial sample of MCM-41 exhibiting multiporosity. Both the delivery mode of hyperpolarized xenon and the stopped-flow system are judged as efficient according to 2D 129Xe self-diffusion and EXSY experiments.

In vivo detection of carnosine and its derivatives using chemical exchange saturation transfer.

PURPOSE: To detect carnosine, anserine and homocarnosine in vivo with chemical exchange saturation transfer (CEST) at 17.2 T. METHODS: CEST MR acquisitions were performed using a CEST-linescan sequence developed in-house and optimized for carnosine detection. In vivo CEST data were collected from three different regions of interest (the lower leg muscle, the olfactory bulb and the neocortex) of eight rats. RESULTS: The CEST effect for carnosine, anserine and homocarnosine was characterized in phantoms, demonstrating the possibility to separate individual contributions by employing high spectral resolution (0.005 ppm) and low CEST saturation power (0.15 µ$$ \mu $$ T). The CEST signature of these peptides was evidenced, in vivo, in the rat brain and skeletal muscle. The presence of carnosine and anserine in the muscle was corroborated by in vivo localized spectroscopy (MRS). However, the sensitivity of MRS was insufficient for carnosine and homocarnosine detection in the brain. The absolute amounts of carnosine and derivatives in the investigated tissues were determined by liquid chromatography-electrospray ionization-tandem mass spectrometry using isotopic dilution standard methods and were in agreement with the CEST results. CONCLUSION: The robustness of the CEST-linescan approach and the favorable conditions for CEST at ultra-high magnetic field allowed the in vivo CEST MR detection of carnosine and related peptides. This approach could be useful to investigate noninvasively the (patho)-physiological roles of these molecules.

Study of syn and anti Xenon-Cryptophanes Complexes Decorated with Aromatic Amine Groups: Chemical Platforms for Accessing New Cryptophanes.

We report the synthesis of C3-symmetric cryptophanes decorated with three aromatic amine groups on the same CTB cap and their interaction with xenon. The relative stereochemistry of these two stereoisomers syn and anti was assessed thanks to the determination of the X-ray structure of an intermediate compound. As previously observed with the tris-aza-cryptophanes analogs anti-1 and syn-2 (J. Org. Chem. 2021, 86, 11, 7648-7658), both compounds anti-5 and syn-6 show a slow in-out exchange dynamics of xenon at 11.7 T. Our work supports the idea that the presence of nitrogen atoms grafted directly onto the cryptophane backbone has a strong impact on the in-out exchange dynamics of xenon whatever their stereochemistry. This result contrasts with the case of other cryptophanes decorated solely with methoxy substituents. Finally, we demonstrate that these new derivatives can be used to design new anti/syn cryptophanes bearing suitable ligands in order to constitute potent 129Xe NMR-based sensors. An example is reported here with the synthesis of the tris-iodo derivatives anti-13 and syn-14 from compounds anti-5 and syn-6.

Are the Physical Properties of Xe@Cryptophane Complexes Easily Predictable? The Case of syn- and anti-Tris-aza-Cryptophanes.

We report the synthesis and optical resolution of C3-symmetrical tris-aza-cryptophanes anti-3 and syn-4, as well as the study of their interaction with xenon via hyperpolarized 129Xe NMR. These molecular cages are close structural analogues of the two well-known cryptophane-A (1; chiral) and cryptophane-B (2; achiral) diastereomers since these new compounds differ only by the presence of three nitrogen atoms grafted onto the same cyclotribenzylene unit. The assignment of their relative (syn vs anti) and absolute configurations was made possible, thanks to the combined use of quantum calculations at the density functional theory level and vibrational circular dichroism spectroscopy. More importantly, our results show that despite the large structural similarities with cryptophane-A (1) and -B (2), these two new compounds show a very different behavior in the presence of xenon in organic solutions. These results demonstrate that prediction of the physical properties of the xenon@cryptophane complexes, only based on structural parameters, remains extremely difficult.

Bimodal Detection of Proteins by 129 Xe NMR and Fluorescence Spectroscopy.

A full understanding of biological phenomena involves sensitive and noninvasive detection. Herein, we report the optimization of a probe for intracellular proteins that combines the advantages of fluorescence and hyperpolarized 129 Xe NMR spectroscopy detection. The fluorescence detection part is composed of six residues containing a tetracysteine tag (-CCXXCC-) genetically incorporated into the protein of interest and of a small organic molecule, CrAsH. CrAsH becomes fluorescent if it binds to the tetracysteine tag. The part of the biosensor that enables detection by means of 129 Xe NMR spectroscopy, which is linked to the CrAsH moiety by a spacer, is based on a cryptophane core that is fully suited to reversibly host xenon. Three different peptides, containing the tetracysteine tag and four organic biosensors of different stereochemistry, are benchmarked to propose the best couple that is fully suited for the in vitro detection of proteins.

Synthesis of Cryptophane-223-Type Derivatives with Dual Functionalization.

In this article, we present the synthesis of new cryptophane-type hosts capable of binding xenon in aqueous media and that may be useful for the development of xenon-based magnetic resonance imaging derivatives. The synthetic route proposed was chosen to facilitate both the introduction of water-solubilizing substituents and the functionalization of the host with a single arm showing recognition properties that constitute two crucial steps. This was made possible by preparing new cryptophane-223 derivatives bearing two different chemical functions that can be easily modified at a later stage. Thus, subsequent reactions allowed the design of a new cryptophane host able to bind zinc or nickel cations. The ability of this molecule to bind cationic species was assessed by calorimetric titration experiments and hyperpolarized 129Xe NMR. The advantages and disadvantages of this approach are discussed.

Role of the Methoxy Groups in Cryptophanes for Complexation of Xenon: Conformational Selection Evidence from 129 Xe-1 H NMR SPINOE Experiments.

We report the laser-polarized 129 Xe and 1 H NMR spectra of a series of cryptophane derivatives that differ only by the number of methoxy groups attached on their benzene rings and the syn or anti arrangement of the linkers (compounds 6 a-s, 9 a-s, 12 a-s). All these compounds bind xenon even though the characteristic signal of the gas encapsulated in the cavity of the cage-molecule cannot always be detected. Interestingly, the exchange dynamics of xenon strongly depends on the degree of substitution and is different from that of the cryptophane derivatives studied previously. In solution, the 1 H NMR spectra of these derivatives show the presence of different conformations in a slow exchange regime that can be explained by a decrease of the flexibility of their skeleton. Thanks to 129 Xe-1 H dipolar cross-relaxation (SPINOE) spectra, we demonstrate that a single conformation present in solution can bind xenon.

Bimodal fluorescence/129Xe NMR probe for molecular imaging and biological inhibition of EGFR in Non-Small Cell Lung Cancer.

Although Non-Small Cell Lung Cancer (NSCLC) is one of the main causes of cancer death, very little improvement has been made in the last decades regarding diagnosis and outcomes. In this study, a bimodal fluorescence/129Xe NMR probe containing a xenon host, a fluorescent moiety and a therapeutic antibody has been designed to target the Epidermal Growth Factor Receptors (EGFR) overexpressed in cancer cells. This biosensor shows high selectivity for the EGFR, and a biological activity similar to that of the antibody. It is detected with high specificity and high sensitivity (sub-nanomolar range) through hyperpolarized 129Xe NMR. This promising system should find important applications for theranostic use.

Synthesis of Cryptophanes with Two Different Reaction Sites: Chemical Platforms for Xenon Biosensing.

We report the synthesis of new water-soluble cryptophane host molecules that can be used for the preparation of (129)Xe NMR-based biosensors. We show that the cryptophane-223 skeleton can be modified to introduce a unique secondary alcohol to the propylenedioxy linker. This chemical functionality can then be exploited to introduce a functional group that is different from the six chemical groups attached to the aromatic rings. In this approach, the generation of a statistical mixture when trying to selectively functionalize a symmetrical host molecule is eliminated, which enables the efficient large-scale production of new cryptophanes that can be used as chemical platforms ready to use for the preparation of xenon biosensors. To illustrate this approach, two molecular platforms have been prepared, and the ability of these new derivatives to bind xenon has been investigated.

Herbicide impact on the growth and reproduction of characteristic and rare arable weeds of winter cereal fields.

The decline of arable species characteristic of winter cereal fields has often been attributed to different factors related to agricultural intensification but most importantly to herbicide use. Herbicide phytotoxicity is most frequently assessed using short-term endpoints, primarily aboveground biomass. However, short-term sensitivity is usually not sufficient to detect actual effects because plants may or may not recover over time following sublethal herbicide exposures. Therefore, it is important to assess the long-term effects of herbicide applications. Annual species rely on renewable seed production to ensure their persistence; hence, assessment of herbicide sensitivity is more accurately estimated through effects on reproduction. Here we aim to assess the phytotoxicity of two commonly used herbicides: tribenuron and 2,4-D on eight plant species belonging to four families, each with one rare and one more common species. Specifically we examined the pattern of sensitivity using short-term and long-term endpoints (total aboveground biomass, total seed biomass and number of seeds) of these species; we determined the levels of and time to recovery in terms of stem length and fruit number, and assessed whether their rarity relates to their sensitivity to herbicide application. Our results suggest that although differences in herbicide sensitivity are not a direct cause of rarity for all species, it may be an important driver of declining arable plants.

Smart detection of toxic metal ions, Pb2+ and Cd2+, using a 129Xe NMR-based sensor.

An approach for sensitive magnetic resonance detection of metal cations is proposed. Combining the use of hyperpolarized (129)Xe NMR and of a cage-molecule functionalized by a ligand able to chelate different cations, we show that simultaneous detection of lead, zinc, and cadmium ions at nanomolar concentration is possible in short time, thanks to fast MRI sequences based on the HyperCEST scheme.

A half-century analysis of landscape dynamics in southern Québec, Canada.

We studied landscape dynamics for three time periods (<1950, 1965, and 1997) along a gradient of agricultural intensity from highly intensive agriculture to forested areas in southern Québec. Air photos were analyzed to obtain long-term information on land cover (crop and habitat types) and linear habitats (hedgerows and riparian habitats) and landscape metrics were calculated to quantify changes in habitat configuration. Anthropogenic areas increased in all types of landscapes but mostly occurred in the highly disturbed cash crop dominated landscape. Perennial crops (pasture and hayfields) were largely converted into annual crops (corn and soybean) between 1965 and 1997. The coalescence of annual crop fields resulted in a more homogeneous agricultural landscape. Old fields and forest cover was consistently low and forest fragmentation remained stable through time in the intensive agriculture landscapes. However, forest cover increased and forest fragmentation receded in the forest-dominated landscapes following farm abandonment and the transition of old fields into forests. Tree-dominated hedgerows and riparian habitats increased in areas with intensive agriculture. Observed changes in land cover classes are related to proximate factors, such as surficial deposits and topography. Agriculture intensification occurred in areas highly suitable for agriculture whereas farm abandonment was observed in poor-quality agriculture terrains. Large-scale conversion of perennial crops into annual crops along with continued urbanization exerts strong pressures on residual natural habitats and their inhabiting wildlife. The afforestation process occurring in the more forested landscapes along with the addition of tree-dominated hedgerows and riparian habitats in the agriculture-dominated landscapes should improve landscape ecological value.

Understanding a host-guest model system through ¹²9Xe†NMR spectroscopic experiments and theoretical studies.

Gaining an understanding of the nature of host-guest interactions in supramolecular complexes involving heavy atoms is a difficult task. Described herein is a robust simulation method applied to complexes between xenon and members of a cryptophane family. The calculated chemical shift of xenon caged in a H2O2 probe, as modeled by quantum chemistry with complementary-orbital, topological, and energy-decomposition analyses, is in excellent agreement with that observed in hyperpolarized (129)Xe NMR spectra. This approach can be extended to other van der Waals complexes involving heavy atoms.

"Clickable" hydrosoluble PEGylated cryptophane as a universal platform for 129Xe magnetic resonance imaging biosensors.

We describe the synthesis of a highly water-soluble cryptophane 1 that can be seen as a universal platform for the construction of (129)Xe magnetic resonance imaging (MRI)-based biosensors. Compound 1 is easily functionalized by Huisgen cycloaddition and exhibits excellent xenon-encapsulation properties. In addition, 1 is nontoxic at the concentrations typically used for hyperpolarized (129)Xe MRI.

Small-area analysis of social inequalities in residential exposure to road traffic noise in Marseilles, France.

BACKGROUND: Few studies have focused on the social inequalities associated with environmental noise despite its significant potential health effects. This study analysed the associations between area socio-economic status (SES) and potential residential exposure to road traffic noise at a small-area level in Marseilles, second largest city in France. METHODS: We calculated two potential road noise exposure indicators (PNEI) at the census block level (for 24-h and night periods), with the noise propagation prediction model CadnaA. We built a deprivation index from census data to estimate SES at the census block level. Locally estimated scatterplot smoothing diagrams described the associations between this index and PNEIs. Since the extent to which coefficient values vary between standard regression models and spatial methods are sensitive to the specific spatial model, we analysed these associations further with various regression models controlling for spatial autocorrelation and conducted sensitivity analyses with different spatial weight matrices. RESULTS: We observed a non-linear relation between the PNEIs and the deprivation index: exposure levels were highest in the intermediate categories. All the spatial models led to a better fit and more or less pronounced reductions of the regression coefficients; the shape of the relations nonetheless remained the same. CONCLUSION: Finding the highest noise exposure in midlevel deprivation areas was unexpected, given the general literature on environmental inequalities. It highlights the need to study the diversity of the patterns of environmental inequalities across various economic, social and cultural contexts. Comparative studies of environmental inequalities are needed, between regions and countries, for noise and other pollutants.

Influence of soil organic matter on the sensitivity of selected wild and crop species to common herbicides.

Current regulatory protocols for assessing herbicide effects on plants rely heavily on the use of crops grown under controlled greenhouse conditions to indicate risks to wild vegetation. Guideline test protocols call for low levels of soil nutrients, approximately 3% organic matter (OM), to test the worst-case scenario for non-target species growing in poor soils. However, species sensitivity to herbicides may be affected by growing conditions, especially soil nutrient levels. The purpose of this study was to assess the impact of high and low soil OM content on the sensitivity of plants to several common agricultural herbicides (glyphosate, chlorimuron ethyl and dicamba). Ten plant species (wild and crop) with high (nitrophilous) or low (non-nitrophilous) affinity for nitrogen were grown under greenhouse conditions in soil with two levels of OM (3 and 9%) and were exposed to seven doses of the three herbicides in concurrent experiments. Results showed that most species were more sensitive to glyphosate under high OM conditions while chlorimuron ethyl and dicamba results were mixed with respect to OM levels. Overall, 15 species-herbicide combinations were more sensitive in high OM soil, while 11 were more sensitive in low OM soil. No clear pattern in sensitivity emerged among nitrophilous and non-nitrophilous species. Several species showed a difference in response at herbicide levels expected to reach non-target habitats adjacent to crop fields through drift (5% commonly and 25% occasionally). In terms of regulatory testing, guidelines may need alteration to allow testing with nutrient levels that more accurately reflect natural environments.

Comparing the sensitivity of geographically distinct Lemna minor populations to atrazine.

The objectives of this study were to compare the sensitivities of field populations and a laboratory culture of a duckweed species (Lemna minor) to the herbicide atrazine using three different endpoints and to determine whether sensitivity to atrazine was affected by past exposure to the herbicide. L. minor cultures were purchased commercially or collected from field sites within an agricultural watershed and exposed to atrazine for 7 days under greenhouse conditions. Populations differed significantly in their sensitivity to atrazine. Biomass was more sensitive than frond number, while chlorophyll fluorescence was not a sensitive endpoint. Overall, the sensitivity of the various populations to atrazine was not strongly related to measures of past exposure to agriculture stressors. Positive correlations between biomass twenty-five percent inhibition concentrations (IC25s), biomass estimated marginal means and in-stream atrazine concentrations were observed, providing evidence that atrazine exposure is linked to a decrease in sensitivity to atrazine. However, IC25s generated for each population were similar, ranging from 19 to 40 and 57 to 92 µg/L atrazine for biomass and frond data respectively, and likely do not represent biologically significant differences in atrazine sensitivity. Given the small range in sensitivity observed between populations, commercial laboratory cultures appear to provide a good estimate of the sensitivity of field populations of L. minor to atrazine and should continue to be used in regulatory phytotoxicity testing.

Vanadium pentoxide phytotoxicity: effects of species selection and nutrient concentration.

Vanadium concentrations in soil can be increased through anthropogenic inputs and can be harmful to plants. A Petri dish experiment was conducted to assess the effect of vanadium toxicity on the germination and survival of the garden lettuce, Lactuca sativa. A second study was conducted in a greenhouse to investigate the influence of species selection and nutrient concentration on the toxicity of vanadium pentoxide to plants. L. sativa and four non-crop native plant species, two grasses (Elymus virginicus and Panicum virgatum) and two broad-leaved species (Lycopus americanus and Prunella vulgaris) were selected. Artificial soil was used in both experiments, and a geometric progression of five vanadium concentrations plus controls was selected for the soil treatments. Results of the Petri dish experiment showed that seedling survival is a less sensitive end point than above-ground dry weight (DW) as measured in the greenhouse experiment. Nutrient level (100, 10, and 1 kg/ha) was found to strongly influence vanadium toxicity in the greenhouse study. At 100 kg/ha, plant tolerance to vanadium was greatest, as indicated by higher no-observed, lowest-observed, and percentage effect concentration values. Results showed that forbs (L. americanus and P. vulgaris) tended to be more sensitive than both the crop (L. sativa) and grasses (E. virginicus and P. virgatum) at high concentrations of vanadium. Soil concentrations resulting in a 25 % decrease in shoot DW were generally less than the Canadian soil quality guideline for vanadium, suggesting that 130 mg/kg may not be protective of the Canadian native plant species used in this study.

Hyperpolarized 129Xe NMR signature of living biological cells.

We show that the differentiation between internal and external compartments of various biological cells in suspension can be made via simple NMR spectra of hyperpolarized (129) Xe. The spectral separation between the signals of (129) Xe in these two compartments is already known for red blood cells, because of the strong interaction of the noble gas with hemoglobin. The observation of two separate peaks in the 200-ppm region can be seen with both eukaryotic and prokaryotic cells, some of which are not known to contain paramagnetic proteins in large quantities. Using different experiments in which the cells are lysed, swell or are blocked in G2 phase, we demonstrate that the low-field-shifted peak observed corresponds to xenon in the aqueous pool inside the cells and not in the membranes. The presence of this additional peak is a clear indication of cell integrity, and its integration allows the quantification of the total cell volume. The relaxation time of intracellular xenon is sufficiently long to open up promising perspectives for cell characterization. The exchange time between the inner and outer cell compartments (on the order of 30 ms) renders possible the targeting of intracellular receptors, whereas the observation of chemical shift variations represents a method of revealing the presence of toxic species in the cells.

Cell uptake of a biosensor detected by hyperpolarized 129Xe NMR: the transferrin case.

For detection of biological events in vitro, sensors using hyperpolarized (129)Xe NMR can become a powerful tool, provided the approach can bridge the gap in sensitivity. Here we propose constructs based on the non-selective grafting of cryptophane precursors on holo-transferrin. This biological system was chosen because there are many receptors on the cell surface, and endocytosis further increases this density. The study of these biosensors with K562 cell suspensions via fluorescence microscopy and (129)Xe NMR indicates a strong interaction, as well as interesting features such as the capacity of xenon to enter the cryptophane even when the biosensor is endocytosed, while keeping a high level of polarization. Despite a lack of specificity for transferrin receptors, undoubtedly due to the hydrophobic character of the cryptophane moiety that attracts the biosensor into the cell membrane, these biosensors allow the first in-cell probing of biological events using hyperpolarized xenon.