Methods for analyzing tellurium imaging mass cytometry data.

Imaging mass cytometry (IMC) is a technique allowing visualization and quantification of over 40 biological parameters in a single experiment with subcellular spatial resolution, however most IMC experiments are limited to endpoint analysis with antibodies and DNA stains. Small molecules containing tellurium are promising probes for IMC due to their cell permeability, synthetic versatility, and most importantly their application to sequential labelling with isotopologous probes (SLIP) experiments. SLIP experiments with tellurium-containing probes allow quantification of intracellular biology at multiple timepoints with IMC. Despite the promise of tellurium in IMC, there are unique challenges in image processing associated with tellurium IMC data. Here, we address some of these issues by demonstrating the removal of xenon background signal, combining channels to improve signal-to-noise ratio, and calculating isotope transmission efficiency biases. These developments add accuracy to the unique temporal resolution afforded by tellurium IMC probes.

A portable ventilator with integrated physiologic monitoring for hyperpolarized 129Xe MRI in rodents.

Hyperpolarized (HP) 129Xe MRI is emerging as a powerful, non-invasive method to image lung function and is beginning to find clinical application across a range of conditions. As clinical implementation progresses, it becomes important to translate back to well-defined animal models, where novel disease signatures can be characterized longitudinally and validated against histology. To date, preclinical 129Xe MRI has been limited to only a few sites worldwide with 2D imaging that is not generally sufficient to fully capture the heterogeneity of lung disease. To address these limitations and facilitate broader dissemination, we report on a compact and portable HP gas ventilator that integrates all the gas-delivery and physiologic monitoring capabilities required for high-resolution 3D hyperpolarized 129Xe imaging. This ventilator is MR- and HP-gas compatible, driven by inexpensive microcontrollers and open source code, and allows for precise control of the tidal volume and breathing cycle in perorally intubated mice and rats. We use the system to demonstrate data acquisition over multiple breath-holds, during which lung motion is suspended to enable high-resolution 3D imaging of gas-phase and dissolved-phase 129Xe in the lungs. We demonstrate the portability and versatility of the ventilator by imaging a mouse model of lung cancer longitudinally at 2 Tesla, and a healthy rat at 7 Tesla. We also report the detection of subtle spectroscopic fluctuations in phase with the heart rate, superimposed onto larger variations stemming from the respiratory cycle. This ventilator was developed to facilitate duplication and gain broad adoption to accelerate preclinical 129Xe MRI research.

Delivery of xenon-containing echogenic liposomes inhibits early brain injury following subarachnoid hemorrhage.

Xenon (Xe), a noble gas, has promising neuroprotective properties with no proven adverse side-effects. We evaluated neuroprotective effects of Xe delivered by Xe-containing echogenic liposomes (Xe-ELIP) via ultrasound-controlled cerebral drug release on early brain injury following subarachnoid hemorrhage (SAH). The Xe-ELIP structure was evaluated by ultrasound imaging, electron microscopy and gas chromatography-mass spectroscopy. Animals were randomly divided into five groups: Sham, SAH, SAH treated with Xe-ELIP, empty ELIP, or Xe-saturated saline. Treatments were administrated intravenously in combination with ultrasound application over the common carotid artery to trigger Xe release from circulating Xe-ELIP. Hematoma development was graded by SAH scaling and quantitated by a colorimetric method. Neurological evaluation and motor behavioral tests were conducted for three days following SAH injury. Ultrasound imaging and electron microscopy demonstrated that Xe-ELIP have a unique two-compartment structure, which allows a two-stage Xe release profile. Xe-ELIP treatment effectively reduced bleeding, improved general neurological function, and alleviated motor function damage in association with reduced apoptotic neuronal death and decreased mortality. Xe-ELIP alleviated early SAH brain injury by inhibiting neuronal death and bleeding. This novel approach provides a noninvasive strategy of therapeutic gas delivery for SAH treatment.

Gas chromatography/mass spectrometry measurement of xenon in gas-loaded liposomes for neuroprotective applications.

RATIONALE: We have produced a liposomal formulation of xenon (Xe-ELIP) as a neuroprotectant for inhibition of brain damage in stroke patients. This mandates development of a reliable assay to measure the amount of dissolved xenon released from Xe-ELIP in water and blood samples. METHODS: Gas chromatography/mass spectrometry (GC/MS) was used to quantify xenon gas released into the headspace of vials containing Xe-ELIP samples in water or blood. In order to determine blood concentration of xenon in vivo after Xe-ELIP administration, 6 mg of Xe-ELIP lipid was infused intravenously into rats. Blood samples were drawn directly from a catheterized right carotid artery. After introduction of the samples, each vial was allowed to equilibrate to 37°C in a water bath, followed by 20 minutes of sonication prior to headspace sampling. Xenon concentrations were calculated from a gas dose-response curve and normalized using the published xenon water-gas solubility coefficient. RESULTS: The mean corrected percent of xenon from Xe-ELIP released into water was 3.87 ± 0.56% (SD, n = 8), corresponding to 19.3 ± 2.8 µL/mg lipid, which is consistent with previous independent Xe-ELIP measurements. The corresponding xenon content of Xe-ELIP in rat blood was 23.38 ± 7.36 µL/mg lipid (n = 8). Mean rat blood xenon concentration after intravenous administration of Xe-ELIP was 14 ± 10 µM, which is approximately 15% of the estimated neuroprotective level. CONCLUSIONS: Using this approach, we have established a reproducible method for measuring dissolved xenon in fluids. These measurements have established that neuroprotective effects can be elicited by less than 20% of the calculated neuroprotective xenon blood concentration. More work will have to be done to establish the protective xenon pharmacokinetic range. Copyright © 2016 John Wiley & Sons, Ltd.

In Vivo Measurement in Pigs of Wash-In Kinetics of Xenon at its Site of Action.

BACKGROUND: Xenon (Xe) in many respects is an ideal anaesthetic agent. Its blood/gas partition coefficient is lower than that of any other anaesthetic, enabling rapid induction of and emergence from anaesthesia. While the whole body kinetics during wash-in of inhalational anaesthesia is well known, data describing the pharmacokinetics of xenon in the cerebral compartment at the site of action are still largely missing. METHODS: In order to illuminate xenon's cerebral pharmacokinetics, we anaesthetised five pigs and measured arterial, mixed- and sagittal sinus-venous blood, as well as end-expiratory gas concentrations of xenon by gas chromatography-mass spectrometry (GCMS) up to 30 minutes after starting the anaesthetic gas mixture. RESULTS: Despite xenon's fast onset of effect the half-time for equilibration between xenon concentration in arterial blood and at the site of action is measured to be 1.49 ± 0.04 minutes versus 3.91 ± 0.1 minutes. Successful loading of xenon in the brain during inhalational anesthesia was accomplished after approximately 15 minutes although the end-expiratory xenon concentration reached a plateau after 7 minutes. Thus cerebral xenon uptake rate is only moderate, xenon fast onset of action being largely due to its extremely fast alveolar uptake. CONCLUSIONS: To ensure safety and precise control during anaesthesia we need a profound knowledge about to what extent the measured end-tidal concentrations reflect the drug concentrations in the target tissue. The results of this study expand our knowledge about the temporal characteristics of xenon´s pharmacokinetics at its site of action and provide the basis for appropriate clinical protocols and experimental designs of future studies.

Minimum alveolar concentration (MAC) for sevoflurane and xenon at normothermia and hypothermia in newborn pigs.

BACKGROUND: Neuroprotection from therapeutic hypothermia increases when combined with the anaesthetic gas xenon in animal studies. A clinical feasibility study of the combined treatment has been successfully undertaken in asphyxiated human term newborns. It is unknown whether xenon alone would be sufficient for sedation during hypothermia eliminating or reducing the need for other sedative or analgesic infusions in ventilated sick infants. Minimum alveolar concentration (MAC) of xenon is unknown in any neonatal species. METHODS: Eight newborn pigs were anaesthetised with sevoflurane alone and then sevoflurane plus xenon at two temperatures. Pigs were randomised to start at either 38.5°C or 33.5°C. MAC for sevoflurane was determined using the claw clamp technique at the preset body temperature. For xenon MAC determination, a background of 0.5 MAC sevoflurane was used, and 60% xenon added to the gas mixture. The relationship between sevoflurane and xenon MAC is assumed to be additive. Xenon concentrations were changed in 5% steps until a positive clamp reaction was noted. Pigs' temperature was changed to the second target, and two MAC determinations for sevoflurane and 0.5 MAC sevoflurane plus xenon were repeated. RESULTS: MAC for sevoflurane was 4.1% [95% confidence interval (CI): 3.65-4.50] at 38.5°C and 3.05% (CI: 2.63-3.48) at 33.5°C, a significant reduction. MAC for xenon was 120% at 38.5°C and 116% at 33.5°C, not different. CONCLUSION: In newborn swine sevoflurane, MAC was temperature dependent, while xenon MAC was independent of temperature. There was large individual variability in xenon MAC, from 60% to 120%.

Single-breath xenon polarization transfer contrast (SB-XTC): implementation and initial results in healthy humans.

PURPOSE: To implement and characterize a single-breath xenon transfer contrast (SB-XTC) method to assess the fractional diffusive gas transport F in the lung: to study the dependence of F and its uniformity as a function of lung volume; to estimate local alveolar surface area per unit gas volume S(A)/V(Gas) from multiple diffusion time measurements of F; to evaluate the reproducibility of the measurements and the necessity of B(1) correction in cases of centric and sequential encoding. MATERIALS AND METHODS: In SB-XTC three or four gradient echo images separated by inversion/saturation pulses were collected during a breath-hold in eight healthy volunteers, allowing the mapping of F (thus S(A)/V(Gas)) and correction for other contributions such as T(1) relaxation, RF depletion and B(1) inhomogeneity from inherently registered data. RESULTS: Regional values of F and its distribution were obtained; both the mean value and heterogeneity of F increased with the decrease of lung volume. Higher values of F in the bases of the lungs in supine position were observed at lower volumes in all volunteers. Local S(A)/V(Gas) (with a mean ± standard deviation of S(A)/V(Gas) = 89 ± 30 cm(-1)) was estimated in vivo near functional residual capacity. Calibration of SB-XTC on phantoms highlighted the necessity for B(1) corrections when k-space is traversed sequentially; with centric ordering B(1) distribution correction is dispensable. CONCLUSION: The SB-XTC technique is implemented and validated for in vivo measurements of local S(A)/V(Gas).

Measurement of blood flow and xenon solubility coefficient in the human liver by xenon-enhanced computed tomography.

PURPOSE: The goal of this work was to develop a method of calculating blood flow and xenon solubility coefficient (λ) in the hepatic tissue by xenon-enhanced computed tomography (Xe-CT) and to demonstrate λ can be used as a measure of fat content in the human liver. METHODS: A new blood supply model is introduced which incorporates both arterial blood and portal venous blood which join and together flow into hepatic tissue. We applied Fick's law to the model. It was theoretically derived that the time course of xenon concentration in the inflow blood (the mixture of the arterial blood and the portal venous blood) can be approximated by a monoexponential function. This approximation made it possible to obtain the time-course change rate (K(I)) of xenon concentration in the inflow blood using the time course of xenon concentration in the hepatic tissue by applying the algorithm we had reported previously. K(I) was used to calculate blood flow and λ for each pixel in the CT image of the liver. Twenty-six patients (49.2 ± 18.3 years) with nonalcoholic steatohepatitis underwent Xe-CT abdominal studies and liver biopsies. Steatosis of the liver was evaluated using the biopsy specimen and its severity was divided into ten grades according to the fat deposition percentage [(severity 1) ≤ 10%, 10 % <(severity 2) ≤ 20%, [ellipsis (horizontal)], 90% < (severity 10) ≤ 100%]. For each patient, blood flow and λ maps of the liver were created, and the average λ value (λ) was compared with steatosis severity and with the CT value ratio of the liver to the spleen (liver∕spleen ratio). RESULTS: There were good correlations between λ and steatosis severity (r = 0.914, P < 0.0001), and between λ and liver∕spleen ratio (r = -0.881, P < 0.0001). Ostwald solubility for xenon in the hepatic tissue (tissue Xe solubility), which is calculated using λ and the hematocrit value of the patient, also showed a good correlation with steatosis severity (r = 0.910, P < 0.0001). λ ranged from 0.86 to 7.81, and tissue Xe solubility ranged from 0.12 to 1.16. This range of solubility is reasonable considering the reported Ostwald solubility coefficients for xenon in the normal liver and in the fat tissue are 0.10 and 1.3, respectively, at 37 °C. The average blood flow value ranged from 15.3 to 53.5 ml∕100 ml tissue∕min. CONCLUSIONS: A method of calculating blood flow and λ in the hepatic tissue was developed by means of Xe-CT. This method would be valid even if portosystemic shunts exist; it is shown that λ maps can be used to deduce fat content in the liver. As a noninvasive modality, Xe-CT would be applicable to the quantitative study of fatty change in the human liver.

Xenon-enhanced dual-energy CT of patients with asthma: dynamic ventilation changes after methacholine and salbutamol inhalation.

OBJECTIVE: The purpose of this study was to evaluate the use of xenon-enhanced dual-energy CT of the chest to assess ventilation changes after methacholine and salbutamol inhalation in subjects with asthma and healthy subjects. SUBJECTS AND METHODS: Twenty-five subjects with asthma and 10 healthy subjects underwent three-phase (basal, after methacholine inhalation, after salbutamol inhalation) xenon-enhanced chest CT. Each phase was composed of wash-in and washout scans. For visual analysis, two radiologists evaluated ventilation defects and gas trapping lobe by lobe on a 10-point scale. Total ventilation defect and gas trapping scores were calculated by adding ventilation defect and gas trapping scores. Xenon and total lung volume were quantified automatically. Total xenon concentration index was calculated as total xenon concentration divided by lung volume. Repeated measures analysis of variance and Student t test were used for comparisons of total ventilation defect score, total gas trapping score, and total xenon concentration index between the two groups. The Friedman test was used for within-group analysis. RESULTS: In the basal state, subjects with asthma had a higher total ventilation defect score (p = 0.004) and higher total gas trapping score (p = 0.05) than did healthy subjects. On washout images, total ventilation defect score, total gas trapping score, and total xenon concentration index after methacholine and salbutamol inhalation were statistically different between the two groups (p < 0.05). However, total xenon concentration index on wash-in images was not significantly different between the two groups. In within-group analysis, total ventilation defect score and total gas trapping score in subjects with asthma and total ventilation defect score in healthy subjects increased significantly after methacholine inhalation and decreased significantly after salbutamol inhalation (p < 0.05). CONCLUSION: Xenon-enhanced chest CT may be a useful technique for visualizing dynamic changes in airflow in response to methacholine and salbutamol inhalation in patients with asthma. Optimization of the protocol for radiation exposure is warranted.

Distribution of hyperpolarized xenon in the brain following sensory stimulation: preliminary MRI findings.

In hyperpolarized xenon magnetic resonance imaging (HP (129)Xe MRI), the inhaled spin-1/2 isotope of xenon gas is used to generate the MR signal. Because hyperpolarized xenon is an MR signal source with properties very different from those generated from water-protons, HP (129)Xe MRI may yield structural and functional information not detectable by conventional proton-based MRI methods. Here we demonstrate the differential distribution of HP (129)Xe in the cerebral cortex of the rat following a pain stimulus evoked in the animal's forepaw. Areas of higher HP (129)Xe signal corresponded to those areas previously demonstrated by conventional functional MRI (fMRI) methods as being activated by a forepaw pain stimulus. The percent increase in HP (129)Xe signal over baseline was 13-28%, and was detectable with a single set of pre and post stimulus images. Recent innovations in the production of highly polarized (129)Xe should make feasible the emergence of HP (129)Xe MRI as a viable adjunct method to conventional MRI for the study of brain function and disease.

[Concept of mechanisms of anesthetic and therapeutic properties of xenon].

The molecular theory of L. Poling is a genius example of scientific prediction, made in the middle of 20th century, when the studies about clathrates and methods of roentgen structured analysis were doing their first steps. The views expressed in this message are some additions on the structure of xenon clathrates, function-free xenon water associates and the use of cameras in the liberated associates of the water molecules, for the process of supramolecular detoxification and attempts to develop this theory more widely, to better understand the mechanisms of xenon anesthesia and treatment for further justification of its therapeutic features as well as for the use of other inert gases (Ar, Kr) in modern medicine.

Xenon as an anesthetic agent.

Discovered in 1898 by British chemists, xenon is a rare gas belonging to the noble gases of the periodic table. Xenon is used in many different ways, from high-intensity lamps to jet propellant, and in 1939, its anesthetic properties were discovered. Xenon exerts its anesthetic properties, in part, through the noncompetitive inhibition of N-methyl-D-aspartate receptors. Currently, xenon is being used primarily throughout Europe; however, the high price of manufacturing and scavenging the noble gas has discouraged more widespread use. As technology in anesthetic delivery improves, xenon is being investigated further as a possible replacement for nitrous oxide as an inhalational agent. This article reviews the anesthetic properties of xenon and current and potential research about the gas.

XTC MRI: sensitivity improvement through parameter optimization.

Xenon polarization Transfer Contrast (XTC) MRI pulse sequences permit the gas exchange of hyperpolarized xenon-129 in the lung to be measured quantitatively. However, the pulse sequence parameter values employed in previously published work were determined empirically without considering the now-known gas exchange rates and the underlying lung physiology. By using a theoretical model for the consumption of magnetization during data acquisition, the noise intensity in the computed gas-phase depolarization maps was minimized as a function of the gas-phase depolarization rate. With such optimization the theoretical model predicted an up to threefold improvement in precision. Experiments in rabbits demonstrated that for typical imaging parameter values the optimized XTC pulse sequence yielded a median noise intensity of only about 3% in the depolarization maps. Consequently, the reliable detection of variations in the average alveolar wall thickness of as little as 300 nm can be expected. This improvement in the precision of the XTC MRI technique should lead to a substantial increase in its sensitivity for detecting pathological changes in lung function.

Recirculation of inhaled xenon does not alter lung CT density.

RATIONALE AND OBJECTIVES: Xenon-enhanced computed tomography (Xe-CT) measures regional ventilation from changes in lung parenchymal CT density during the multibreath washin/washout of inhaled Xe gas. Because Xe is moderately soluble, vascular uptake and redistribution has been proposed as a confounding phenomenon. We propose that the redistribution of Xe via the circulation is negligible, and correction is unwarranted. MATERIALS AND METHODS: Unilateral ventilation with 60% Xe was performed in intubated canines. Whole-lung CT images were obtained at baseline and after 1 and 5 minutes of unilateral Xe ventilation. Comparisons between blocked (B) and Xe ventilated (V) whole lung densities were made. Density of paraspinous muscle and blood (aorta, inferior vena cava) were also compared. RESULTS: The density of lung tissue in the V lungs increased significantly compared to B lungs after 1 minute (B -688.5 +/- 54.3 Hounsfield units [HU] vs. V -535.4 +/- 55.6 HU, P < .05) and 5 minutes (B -689.1 +/- 52.2 HU vs. V -492.9 +/- 89.1 HU, P < .05) of Xe ventilation. The density in the blocked lungs did not significantly change after either 1 or 5 minutes of ventilation with Xe. Although density tended to increase with time in the blood and muscle, the change only reached significance in muscle at 5 minutes. CONCLUSIONS: Five minutes of ventilation with a high concentration of Xe does not cause measurable density changes in the contralateral, unventilated lung. Xe accumulation in muscle tissue limits redistribution. Correction of Xe-CT time series density data may be unnecessary.

Cell effects of xenon in vitro under hypothermal conditions.

The effects of Xe on cell viability and redox balance in the culture of Wistar rat thymocytes were studied in vitro during 24-h storage under hypothermic conditions. The results indicate that after bubbling of cell suspensions (5 x 10(6) cell/ml, 4 ml medium), the weight of Xe in flasks was 15-43 mg, whereas in cell-free medium no weight increment due to gas accumulation in the system was detected. The content of Xe in cell suspension slightly decreased over 24-h culturing at ambient temperature (by 10% of initial level). Xenon significantly improved cell survival during thermal exposure of all modes. The maximum cytoprotective effect of Xe was observed under rigorous thermal conditions associated with significant cell death without chemical protectors (3 degrees C, -35 degrees C). The effect of Xe was less pronounced at mild temperatures (23-37 degrees C) or in the presence of chemical protectors (-35 degrees C with dimethylsulfoxide). The mechanisms of the effect of the inert gas are determined by its antioxidant or prooxidant action. The capacity of Xe to improve cell survival under hypothermic conditions can be used for the development of new methods for transportation and storage of cell material.

Inert gases as the future inhalational anaesthetics?

Of all the inert gases, only xenon has considerable anaesthetic properties under normobaric conditions. Its very low blood/gas partition coefficient makes induction of and emergence from anaesthesia more rapid compared with other inhalational anaesthetics. In experimental and clinical studies the safety and efficiency of xenon as an anaesthetic has been demonstrated. Xenon causes several physiological changes, which mediate protection of the brain or myocardium. The use of xenon might therefore be beneficial in certain clinical situations, as in patients at high risk for neurological or cardiac damage.

Dynamic NMR spectroscopy of hyperpolarized (129)Xe in human brain analyzed by an uptake model.

Hyperpolarized (129)Xe (HpXe) NMR not only holds promise for functional lung imaging, but for measurements of tissue perfusion as well. To investigate human brain perfusion, several time-series of (129)Xe MR spectra were recorded from one healthy volunteer after HpXe inhalation. The time-dependent amplitudes of the MR spectra were analyzed by using a compartment model for xenon uptake modified to account for the loss of (129)Xe polarization due to RF-excitation and for the breathhold technique used in the experiments. This analysis suggests that the resonances detected at 196.5 +/- 1 ppm and 193 +/- 1 ppm originate from HpXe dissolved in gray and white matter, respectively, and that T(1) relaxation times of HpXe are different in gray and white matter (T(1g) > T(1w)).