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Direct 3D bioprinting of bioreactors containing microorganisms embedded inside hydrogel structures is a promising strategy for biotechnological applications. Nevertheless, microporous hydrogel networks hinder the supply of nutrients and oxygen to the cell and limit cell migration and proliferation. To overcome this drawback, we developed a feedstock for 3D bioprinting structures with hierarchical porosity. The feedstock is based on a highly particle-filled alumina/alginate nanocomposite gel with immobilized E. coli bacteria with the protein ovalbumin acting as foaming agent. The foamed nanocomposite is shaped into a porous mesh structure by 3D printing. The pore radius diameters inside the non-printed, non-foamed nanocomposite structure are below 10 µm, between 10 and 500 µm in the albumin-stabilized foam and with additional pores in the range of 0.5 and 1 mm in the printed mesh structure. The influence of albumin on the bubbles and hence pore formation was analyzed by means of interfacial shear rheology and porosity measurements with X-ray microtomography (µCT). Furthermore, averaged diffusion coefficients of water in printed and non-printed samples with different albumin concentrations were recorded using nuclear magnetic resonance (NMR) tomography to assess the water content in the porous structure. Moreover, the effective viability and accessibility of embedded E. coli cells were analyzed for various material compositions. Here, the addition of albumin induced bacterial growth and the porosity increased the effective viability of the embedded bacteria, most likely because of enhanced accessibility of the cells. The experimental results demonstrate the potential of this approach for producing macroscopic bioactive materials with complex 3D geometries as a platform for novel applications in bioprocessing.

The use of magnetic resonance imaging can contribute to a better understanding of limiting processes occurring in electroactive biofilms especially inside opaque porous electrodes.

In reaction engineering, it is often that overcoming transport limitations improves reactor performance. This requires detailed analyses of transport phenomena in the catalytic beds. Nuclear magnetic resonance velocimetry (MRV) measurements have been utilized for analyzing mass transport of gas flows within opaque monoliths. Comparisons to full-field computational fluid dynamics (CFD) simulations, however, show significant deviations. In this study, polyethylene glycol (PEG) and 3D-printed monoliths including one open-cell foam (OCF) and one honeycomb were used to demonstrate that both operating fluid and monolith morphology influence the achievable signal-to-noise ratio and resolution of NMR data. The velocity profiles measured by MRV in OCF agreed well with full-field CFD simulations with ± 5% deviation. In addition, the similarity between the simulated and experimental velocity fields was quantified by the similarity index, which is 1 for identical images. A mean value of 0.83 was determined for a 10 PPI OCF. Thus, using PEG as the operating fluid and a 10 PPI OCF allows to improve both spatial resolution by 34% and the quality of agreement by 13 percentage points compared to the published results of gas velocimetry within 20 PPI OCF. We further identified and quantified possible sources of deviation between CFD and MRV velocity fields. By limiting our analysis to velocities higher than 45% of the maximum velocity, we could achieve similarity indices of 0.95–0.99.

Chemical exchange saturation transfer (CEST) at about 2.8 ppm downfield from water is characterized besides other compounds by exchanging amine protons of relatively high concentration amino acids and is determined by several physiological (pH, T) and experimental (B0 , B1 , tsat ) parameters. Although the weighting of the CEST effect observed in vivo can be attributed mainly to one compound depending on the organism and organ, there are still several other amino acids, proteins and molecules that also contribute. These contributions in turn exhibit dependences and thus can lead to possible misinterpretation of the measured changes in the CEST effect. With this in mind, this work aimed to determine the exchange rates of six important amino acids as a function of pH and temperature, and thus to create multi-pool models that allow the accurate analysis of the CEST effect concerning different physiological and experimental parameters for a wide variety of organisms. The results show that small changes in the above parameters have a significant impact on the CEST effect at about 2.8 ppm for the chosen organisms, i.e. the human brain (37 °C) and the brain of polar cod (1.5 °C), furthermore, the specificity of the CEST effect observed in vivo can be significantly affected. Based on the exchange rates ksw (pH, T) determined for six metabolites in this study, it is possible to optimize the intensity and the specificity for the CEST effect of amino acids at about 2.8 ppm for different organisms with their specific physiological characteristics. By adjusting experimental parameters accordingly, this optimization will help to avoid possible misinterpretations of CEST measurements. Furthermore, the multi-pool models can be utilized to further optimize the saturation.

Recently, in situ studies using nuclear magnetic resonance (NMR) have shown the possibility to monitor local transport phenomena of gas-phase reactions inside opaque structures. Their application to heterogeneously catalyzed reactions remains challenging due to inherent temperature and pressure constraints. In this work, an NMR-compatible reactor was designed, manufactured, and tested, which can endure high temperatures and increased pressure. In temperature and pressure tests, the reactor withstood pressures up to 28 bars at room temperature and temperatures over 400 °C and exhibited only little magnetic shielding. Its applicability was demonstrated by performing the CO2 methanation reaction, which was measured operando for the first time by using a 3D magnetic resonance spectroscopic imaging sequence. The reactor design is described in detail, allowing its easy adaptation for different chemical reactions and other NMR measurements under challenging conditions.

In order to investigate drying mechanisms at different stages, the distribution of water within the ceramic green bodies at different scales has been examined. The experimental measurements, using a simple weighing technique and Magnetic Resonance Imaging (MRI), show that during the first stage of drying involving shrinkage the material is constituted of uniquely solid and water with no gradient in water content within the sample. Then, during the second stage of drying, significant differences of water content as a function of position appear. As a complement, at the grain scale, observations using environmental scanning electron microscopy were made giving useful information on the solid–liquid–gas interfaces in the near surface part of the green body. Finally, the gradients in the water distribution were exploited to make a simple estimate of the diffusion coefficient of water with its dependence on the moisture content.

In situ thermometry of catalytic gas phase reactions allows the determination of temperature profiles in catalyst beds. In NMR imaging systems used for measuring the chemical composition of species in model reactors, temperature measurements by NMR spectroscopy are technically challenging and confined to a rather low temperature range. In this study, an optimized NMR in situ technique is proposed, which will allow the determination of the temperature distribution in highly exothermic reactions on structured catalysts. Diffusion weighted magnetic resonance imaging (DW-MRI) was successfully applied as an alternative method for temperature measurements commonly performed by chemical shift measurements using ethylene glycol. DW-MRI applied with different diffusion sensitizing gradients allows high-resolution imaging of the temperature dependent diffusion coefficient, without the need for high spatial homogeneity of the magnetic field. Using 3D DW-MRI on ethylene glycol, glycerol, and the temperature stable ionic liquid Pyr13 [TFSI] (decomposition temperature of 400 °C) as NMR thermometers, measurements were performed in a temperature range from 20 to 160 °C. The proposed method can be used in reaction engineering approaches performed in NMR systems.

In today's industrial processes reactions in dispersed gas–liquid systems are of major importance. Many products originate from gas–liquid reactions inside the bubble wake, acting as a mixing zone. High reaction yields are mainly influenced by the hydrodynamics within these zones. However, undisturbed hydrodynamic measurements of low viscous systems inside the bubble wakes are lacking. In this work we report on non-invasive MRI of gas–liquid Taylor flows. A detailed explanation of the developed MRI setup and sequence is given.

The neurochemical information provided by proton magnetic resonance spectroscopy (MRS) or MR spectroscopic imaging (MRSI) can be severely compromised if strong signals originating from brain water and extracranial lipids are not properly suppressed. The authors of this paper present an overview of advanced water/lipid-suppression techniques and describe their advantages and disadvantages. Moreover, they provide recommendations for choosing the most appropriate techniques for proper use. Methods of water signal handling are primarily focused on the VAPOR technique and on MRS without water suppression (metabolite cycling). The section on lipid-suppression methods in MRSI is divided into three parts. First, lipid-suppression techniques that can be implemented on most clinical MR scanners (volume preselection, outer-volume suppression, selective lipid suppression) are described. Second, lipid-suppression techniques utilizing the combination of k-space filtering, high spatial resolutions and lipid regularization are presented. Finally, three promising new lipid-suppression techniques, which require special hardware (a multi-channel transmit system for dynamic B1+ shimming, a dedicated second-order gradient system or an outer volume crusher coil) are introduced.

In the last decade, in-situ studies using nuclear magnetic resonance (NMR) showed the possibility to monitor local transport phenomena of gas-phase reactions inside opaque structures. While spatial species concentration is known to be accessible operando during heterogeneously catalyzed gas-phase reactions, the spatial acquisition of gas temperatures has proven to be challenging. So far, temperature information is gathered by auxiliary liquid-filled capillaries with temperature-dependent NMR-properties. Here, we present a method for spatially resolved temperature and density measurements of gases using the temperature dependence of the signal amplitude and the longitudinal relaxation time (T1). In order to support the proposed method, temperature measurements of methane-filled tubes are carried out and compared to state-of-the-art temperature measurements using glycerol-filled capillaries. To demonstrate the applicability of the method during heterogeneously catalyzed gas-phase reactions, we measured the temperature operando inside a catalytically active honeycomb structure at standard pressure. The results show that direct temperature measurements of gases are possible, even though the low pressure inside the methane-filled tubes led to a low signal-to-noise ratio which compromised the accuracy of the data yielding relative errors of up to 7 %. This study focusses on temperature measurements of pure methane only. When applied to mixtures of gases, however, the proposed method has the potential to measure gas temperature and concentration at the same time.

Abstract Pattern of fluid flow through open-cell foams is important because of its influence on the performance of processes such as filtration, adsorption and heterogeneous catalysis that make use of such foams. So far, however, the experimental verification of velocity profiles obtained by computational fluid dynamics (CFD) simulation was insufficient. Here, the effect of morphology of ceramic foams on local gas flow patterns is observed via the noninvasive magnetic resonance velocimetry (MRV) technique. In order to cross-validate the simulations with the experimental flow mapping results, micro-computed tomography (µCT) data of the entire foams were used for generating the computational network required for 3D CFD simulations of velocity fields within the pores. The results of CFD simulations and MRV measurements of gas flow showed a remarkable agreement with deviations mainly below 10 percent if the whole foam structure was utilized in CFD simulations. The qualitative and quantitative agreement between CFD and MRV results underlines the reliability of CFD simulations that are based on µCT data and underpins the capability of NMR-based measurements for in situ velocity measurements. Graphic abstract

The neurochemical information provided by proton magnetic resonance spectroscopy (MRS) or MR spectroscopic imaging (MRSI) can be severely compromised if strong signals originating from brain water and extracranial lipids are not properly suppressed. The authors of this paper present an overview of advanced water/lipid-suppression techniques and describe their advantages and disadvantages. Moreover, they provide recommendations for choosing the most appropriate techniques for proper use. Methods of water signal handling are primarily focused on the VAPOR technique and on MRS without water suppression (metabolite cycling). The section on lipid-suppression methods in MRSI is divided into three parts. First, lipid-suppression techniques that can be implemented on most clinical MR scanners (volume preselection, outer-volume suppression, selective lipid suppression) are described. Second, lipid-suppression techniques utilizing the combination of k-space filtering, high spatial resolutions and lipid regularization are presented. Finally, three promising new lipid-suppression techniques, which require special hardware (a multi-channel transmit system for dynamic B

For experimental investigations of colloid retention in porous media, also denoted as deep bed filtration, X-ray computed microtomography (µCT) has become a basic tool within the last decade. On the one hand, µCT can spatially resolve particle deposition at discrete points of filtration time. On the other hand, the topological information of the porous media including the porosity and the pore size distribution can be obtained. Aside from structural parameters, the velocity field of the fluid within the pores, which cannot be measured by means of µCT, plays an important role in the underlying mechanisms of particle transport and immobilization. In a given structure, a high flow rate will result in increased velocity gradients as well as increased shear forces compared to a lower flow rate. High shear forces are in turn unfavorable for particle deposition. Another imaging modality, magnetic resonance velocimetry (MRV), is capable of quantifying the desired velocity maps. We demonstrate an experimental approach that combines both, MRV and µCT. In contrast to the majority of other investigations about colloid retention, the porous media investigated in this work are monolithic foam-like structures. The evaluation of colloid deposition in those monolithic filters is based on analyzing individual pores. Particle deposition in a pore is expressed by the volumetric fraction of particles while the pore flow is described by the Reynolds number. Results indicate that pores with high Reynolds numbers are not among the pores with the highest or lowest volume fraction of particles for a given time. The particle volume fraction in pores with low Reynolds numbers is mainly a function of the axial position of the pore.

A heterogeneously catalyzed gas phase reaction process was characterized regarding temperature and concentration profiles by means of three dimensional (3D) 1 H magnetic resonance spectroscopic imaging (MRSI), using the exothermal ethylene hydrogenation as an example. Here, temperature mapping was achieved by using specifically designed thermometers filled with ethylene glycol. The impact of heat and mass transfer on the process performance was investigated by using two different monolithic catalysts with completely different heat and mass transfer characteristics: a regularly structured honeycomb monolith and a irregularly structured open-cell foam packing. The influence of these characteristics on the reaction zones within the monolithic catalysts was demonstrated by simulations that were based on 2D reactor models. To evaluate the applicability of temperature and concentration mapping by 1 H MRSI for model validation, a predictive two dimensional model of the process was applied. The resulting simulations of temperature profiles and concentration distributions were in very good agreement with the experimental data with deviations below 9%. Conventional mass spectroscopic measurements provided further evidence of the accuracy of 3D MRSI measurements as well as the 2D reactor model.

Objectives: Temperature dependent chemical shifts of important brain metabolites measured by localised 1H MRS were investigated to test how the use of incorrect prior knowledge on chemical shifts impairs the quantication of metabolite concentrations. Materials and methods: Phantom measurements on solutions containing 11 metabolites were performed on a 7 T scanner between 1 and 43 °C. The temperature dependence of the chemical shift differences was fitted by a linear model. Spectra were simulated for di erent temperatures and analysed by the AQSES program (jMRUI 5.2) using model functions with chemical shift values for 37 °C. Results: Large differences in the temperature dependence of the chemical shift differences were determined with a maximum slope of about ±7.5 × 10−4 ppm/K. For 32–40 °C, only minor quantification errors resulted from using incorrect chemical shifts, with the exception of Cr and PCr. For 1–10 °C considerable quantification errors occurred if the temperature dependence of the chemical shifts was neglected. Conclusion: If 1H MRS measurements are not performed at 37 °C, for which the published chemical shift values have been determined, the temperature dependence of chemical shifts should be considered to avoid systematic quantification errors, particularly for measurements on animal models at lower temperatures.

A micro-scale analysis of mass transport in ceramic foams that are used as catalyst supports in gas phase reactions is of high interest. Although the effects of flow rate and foam parameters on the radial and axial dispersion are known for liquid flows, no pore-scale experimental analysis has been yet reported to correlate the mechanical and diffusional dispersion of gas flows to the geometry of open-cell foams. Here, a spatially resolved Pulsed Field Gradient NMR method is applied to determine dispersion coefficients of thermally polarized gas along axial and transversal directions of open-cell foams. The comparative study of three commercial foam samples with different morphologies shows the effect of open porosity, window size, and flow rate on gas dispersion. Additionally, the influence of mechanical and diffusional dispersion at each flow rate is investigated for individual samples. By observing the transition from diffusional dispersion to mechanically driven dispersion of gas, it is found that diffusional dispersion plays an important role, even at higher flow rates after a transition from Darcy to Darcy-Forchheimer regime occurs. The measured values for dispersion coefficients of methane can be directly used in pseudo-heterogeneous models for the methanation reaction.

Chemical exchange saturation transfer (CEST) from taurine to water (TauCEST) can be used for in vivo mapping of taurine concentrations as well as for measurements of relative changes in intracellular pH (pHi ) at temperatures below 37°C. Therefore, TauCEST offers the opportunity to investigate acid-base regulation and neurological disturbances of ectothermic animals living at low temperatures, and in particular to study the impact of ocean acidification (OA) on neurophysiological changes of fish. Here, we report the first in vivo application of TauCEST imaging. Thus, the study aimed to investigate the TauCEST effect in a broad range of temperatures (1-37°C) and pH (5.5-8.0), motivated by the high taurine concentration measured in the brains of polar fish. The in vitro data show that the TauCEST effect is especially detectable in the low temperature range and strictly monotonic for the relevant pH range (6.8-7.5). To investigate the specificity of TauCEST imaging for the brain of polar cod (Boreogadus saida) at 1.5°C simulations were carried out, indicating a taurine contribution of about 65% to the in vivo expected CEST effect, if experimental parameters are optimized. B. saida was acutely exposed to three different CO2 concentrations in the sea water (control normocapnia; comparatively moderate hypercapnia OAm = 3300 μatm; high hypercapnia OAh = 4900 μatm). TauCEST imaging of the brain showed a significant increase in the TauCEST effect under the different CO2 concentrations of about 1.5-3% in comparison with control measurements, indicative of changes in pHi or metabolite concentration. Consecutive recordings of 1 H MR spectra gave no support for a concentration induced change of the in vivo observed TauCEST effect. Thus, the in vivo application of TauCEST offers the possibility of mapping relative changes in pHi in the brain of polar cod during exposure to CO2 .

Chemical exchange saturation transfer (CEST) from glutamate to water (GluCEST) is a powerful tool for mapping glutamate concentration and intracellular pH. GluCEST could also be helpful to understand the physiology of lower aquatic vertebrates and invertebrates. Therefore, this study aimed to investigate the GluCEST effect and the exchange rate ksw from amine protons of glutamate to water in a broad range of temperatures (1-37°C) and pH (5.5-8.0). Z-spectra were measured from glutamate solutions at different pH values and temperatures and analysed by numerically solving the Bloch-McConnell equation. As expected, a strong dependence of the GluCEST effect and the determined ksw values on pH and temperature was observed. In addition, a strong dependence of the GluCEST effect on phosphate buffer concentration was confirmed. The in vitro data show that GluCEST is detectable in the whole temperature range, even at 1°C. An interpolation function for the exchange rate ksw was determined for the considered range of temperatures and pH values, showing a bijective relation between the exchange rate and pH at a given temperature. To investigate the specificity of GluCEST imaging at low temperatures, the CEST effect was investigated for several metabolites relevant for CEST imaging of the brain. As an example, the contribution of GluCEST to the total CEST effect at 3 ppm was estimated for zebrafish (Danio rerio). It is shown that also at lower temperatures glutamate is the major contributor to the total CEST effect, particularly if the experimental parameters are optimized.

In situ analysis of heterogeneously catalyzed gas phase reaction systems is becoming a valuable aid to their modeling and optimization. The commonly applied methods are either invasive, do not provide spatial information or are not applicable for optically inaccessible systems. This work investigates the possibility to use NMR imaging to study gas phase reaction processes in situ, spatially resolved and non-invasively. A multislice NMR spectroscopic imaging pulse sequence, which was optimized to realize ultrashort echo time TE, was employed to study the ethylene hydrogenation reaction in an NMR-compatible packed bed flow reactor. The catalyst bed, containing inactive γ -Al 2 O 3 pellets and Pt-Al 2 O 3 pellets, was subdivided into several sections in order to identify reaction zones that depend on initial conditions. Spatial mapping of the chemical composition was demonstrated on the basis of two experiments with varying initial volume flow and ethylene conversion. The inlet and outlet temperature of the catalyst bed was simultaneously detected by analyzing the spectra of inserted glycol capsules. The resulting spatial shift of the reactive zones in both experiments could be proven by the spatially resolved concentration measurements and the temperature measurements. The locations of single active catalyst pellets were also detectable by the same measure. The quantitative results of product gas composition of both experiments were in good agreement with accompanying mass spectrometric measurements. The results demonstrate the applicability of NMR imaging methods to investigate gas phase reaction processes and can help to establish these methods as a standard tool to map chemical transformations in gas flow reactors.

Diabetes diagnostic therapy and research would strongly benefit from noninvasive accurate imaging of the functional β-cells in the pancreas. Here, we developed an analysis of functional β-cell mass (BCM) by measuring manganese (Mn2+) uptake kinetics into glucose-stimulated β-cells by T1-weighted in vivo Mn2+-mediated MRI (MnMRI) in C57Bl/6J mice. Weekly MRI analysis during the diabetes progression in mice fed a high-fat/high-sucrose diet (HFD) showed increased Mn2+-signals in the pancreas of the HFD-fed mice during the compensation phase, when glucose tolerance and glucose-stimulated insulin secretion (GSIS) were improved and BCM was increased compared with normal diet–fed mice. The increased signal was only transient; from the 4th week on, MRI signals decreased significantly in the HFD group, and the reduced MRI signal in HFD mice persisted over the whole 12-week experimental period, which again correlated with both impaired glucose tolerance and GSIS, although BCM remained unchanged. Rapid and significantly decreased MRI signals were confirmed in diabetic mice after streptozotocin (STZ) injection. No long-term effects of Mn2+ on glucose tolerance were observed. Our optimized MnMRI protocol fulfills the requirements of noninvasive MRI analysis and detects already small changes in the functional BCM.

Chemical exchange saturation transfer (CEST) from taurine to water (TauCEST) can be used for in vivo mapping of taurine concentrations as well as for measurements of relative changes in intracellular pH (pH

Temperature dependent chemical shifts of important brain metabolites measured by localisedPhantom measurements on solutions containing 11 metabolites were performed on a 7 T scanner between 1 and 43 °C. The temperature dependence of the chemical shift differences was fitted by a linear model. Spectra were simulated for different temperatures and analysed by the AQSES program (jMRUI 5.2) using model functions with chemical shift values for 37 °C.Large differences in the temperature dependence of the chemical shift differences were determined with a maximum slope of about ±7.5 × 10If

MR velocimetry of liquid flow in opaque porous filters may play an important role in better understanding the mechanisms of deep bed filtration. With this knowledge, the efficiency of separating the suspended solid particles from the vertically flowing liquid can be improved, and thus a wide range of industrial applications such as wastewater treatment and desalination can be optimized. However, MR velocimetry is challenging for such studies due to the low velocities, the severe B0 inhomogeneity in porous structures, and the demand for high spatial resolution and an appropriate total measurement time during which the particle deposition will change velocities only marginally. In this work, a modified RARE-based MR velocimetry method is proposed to address these issues for velocity mapping on a deep bed filtration cell. A dedicated RF coil with a high filling factor is constructed considering the limited space available for the vertical cell in a horizontal MR magnet. Several means are applied to optimize the phase contrast RARE MRI pulse sequence for accurately measuring the phase contrast in a long echo train, even in the case of a low B1 homogeneity. Two means are of particular importance. One uses data acquired with zero flow to correct the phase contrast offsets from gradient imperfections, and the other combines the phase contrast from signals of both odd and even echoes. Results obtained on a 7T preclinical MR scanner indicate that the low velocities in the heterogeneous system can be correctly quantified with high spatial resolution and an adequate total measurement time, enabling future studies on flow during the filtration process.

Monolithic reactor concepts are currently intensively discussed in the literature. For the oxidative coupling of methane relying on a balance of surface and gas phase reactions, such concepts have previously been claimed to have beneficial effects with respect to obtainable C2 yields. In order to verify the superior performance in the case of a foam catalyst, ceria and samaria foams were fabricated by a direct foaming process. In both cases, mechanically stable, homogeneous open-cell foams were obtained as revealed by 3D magnetic resonance imaging, Hg-porosimetry, and scanning electron microscopy. As a characteristic feature of the introduced foaming methodology the process resulted in bimodal pore size distributions, ensuring low pressure drops on the one hand and sufficiently large surface areas on the other hand. Oxidative coupling of methane was carried out over the samaria foams. It was possible to obtain C2 yields that were indeed higher than those obtained with the samaria powder, in contrast to honeycomb monoliths previously studied in the literature.

Measuring spatially resolved concentration distributions in gas phase reaction systems is an important tool to validate simulation calculations, improve the understanding of transport processes within the catalyst, and identify potentials for improvements of monolithic catalyst supports. The commonly used measurement methods for such opaque systems are invasive and, thus, might be misleading due to alteration of the system. To overcome this issue, a 3D magnetic resonance spectroscopic imaging (MRSI) method was developed and implemented on a 7-Tesla NMR imaging system to map the concentration distributions within opaque monolithic catalysts using the ethylene hydrogenation process as case study. The reaction was catalyzed by a coated sponge packing or a honeycomb monolith within an NMR compatible packed bed reactor. Temperatures at the inlet and the outlet of the catalyst beds were simultaneously determined by analyzing the spectra of inserted ethylene glycol filled glass capsules. Steady state concentration profiles and temperature levels were measured at different reaction conditions. In order to prove the plausibility of the measured spatial distributions of compound concentrations, the experimental results were compared to a 1D model of the reactor based on kinetic data from literature. Furthermore, a comparison with integral concentration measurements using a mass spectrometer demonstrated deviations below 5%. The results show that 3D MRSI is a valuable and reliable tool to non-invasively measure spatially resolved process parameters within optically and/or mechanically inaccessible structured monolithic catalyst supports, even if only standard thermal polarization is exploited and the use of expensive and technically challenging signal enhancement techniques (hyperpolarization) is avoided. We expect that 3D MRSI can pave the way toward deeper insight into the interactions between catalyst, catalyst support, and gas phase.

The use of water suppression for in vivo proton MR spectroscopy diminishes the signal intensities from resonances that undergo magnetization exchange with water, particularly those downfield of water. To investigate these exchangeable resonances, an inversion transfer experiment was performed using the metabolite cycling technique for non-water-suppressed MR spectroscopy from a large brain voxel in 11 healthy volunteers at 3.0 T. The exchange rates of the most prominent peaks downfield of water were found to range from 0.5 to 8.9 s(-1), while the T(1) relaxation times in absence of exchange were found to range from 175 to 525 ms. These findings may help toward the assignments of the downfield resonances and a better understanding of the sources of contrast in chemical exchange saturation transfer imaging.

Chemical exchange saturation transfer (CEST) from glutamate to water (GluCEST) is a powerful tool for mapping glutamate concentration and intracellular pH. GluCEST could also be helpful to understand the physiology of lower aquatic vertebrates and invertebrates. Therefore, this study aimed to investigate the GluCEST effect and the exchange rate ksw from amine protons of glutamate to water in a broad range of temperatures (1-37°C) and pH (5.5-8.0). Z-spectra were measured from glutamate solutions at different pH values and temperatures and analysed by numerically solving the Bloch-McConnell equation. As expected, a strong dependence of the GluCEST effect and the determined ksw values on pH and temperature was observed. In addition, a strong dependence of the GluCEST effect on phosphate buffer concentration was confirmed. The in vitro data show that GluCEST is detectable in the whole temperature range, even at 1°C. An interpolation function for the exchange rate ksw was determined for the considered range of temperatures and pH values, showing a bijective relation between the exchange rate and pH at a given temperature. To investigate the specificity of GluCEST imaging at low temperatures, the CEST effect was investigated for several metabolites relevant for CEST imaging of the brain. As an example, the contribution of GluCEST to the total CEST effect at 3 ppm was estimated for zebrafish (Danio rerio). It is shown that also at lower temperatures glutamate is the major contributor to the total CEST effect, particularly if the experimental parameters are optimized.

Modern NMR imaging systems used for biomedical research are equipped with B0 gradient systems with strong maximum gradient strength and short switching time enabling (1)H NMR measurements of samples with very short transverse relaxation times. However, background signal originating from non-optimized RF coils may hamper experiments with ultrashort delays between RF excitation and signal reception. We demonstrate that two simple means, outer volume suppression and the use of shaped B0 fields produced by higher-order shim coils, allow a considerable suppression of disturbing background signals. Thus, the quality of NMR images acquired at ultrashort or zero echo time is improved and systematic errors in quantitative data evaluation are avoided. Fields of application comprise MRI with ultrashort echo time or relaxation time analysis, for both biomedical research and characterizing porous media filled with liquids or gases.

A methodological development for quantitative short-echo-time (TE) in vivo proton MR spectroscopy (MRS) without water suppression (WS) is described that integrates experimental and software approaches. Experimental approaches were used to eliminate frequency modulation sidebands and first-order phase errors. The dominant water signal was modeled and extracted by the matrix pencil method (MPM) and was used as an internal reference for absolute metabolite quantification. Spectral fitting was performed by combining the baseline characterization by a wavelet transform (WT)-based technique and time-domain (TD) parametric spectral analysis using full prior knowledge of the metabolite model spectra. The model spectra were obtained by spectral simulation instead of in vitro measurements. The performance of the methodology was evaluated by Monte Carlo (MC) studies, phantom measurements, and in vivo measurements on rat brains. More than 10 metabolites were quantified from spectra measured at TE = 20 ms on a 4.7 T system.

An experimental method is described to eliminate frequency modulation sidebands in localized in vivo proton MR spectra acquired without water suppression. Based on the fact that the phases of the sideband artifact signals are coherent with the phases of the gradient pulses, the proposed method measures and combines signals with sequences using opposite gradients to cancel the sidebands. Factors effecting the results of the cancellation are investigated. Optimized cancellation is achieved by trimming the refocusing gradients, optimizing the spoiler gradients, or replacing the spoiler gradients with a 16-step RF phase cycling. The performance of the method is demonstrated by phantom and in vivo experiments on the healthy rat brain. Magn Reson Med 51:602–606, 2004. © 2004 Wiley-Liss, Inc.

Modified point-resolved spectroscopy (PRESS) sequences for single voxel spetroscopy (MRS) and spectroscopic imaging (SI) with very short echo time (T E ) are described using asymmetric radio-frequency (RF) pulses as well as an optimized design and timing of the PRESS sequence. The proposed sequences were implemented on a standard 4.7 T imaging system yielding a T E of 6.0 ms only. Simulations and experimental data measured on phantoms and the rat brain in vivo are presented for MRS and SI showing a high signal-to-noise ratio and hardly any phase distortions caused by J-coupling.

Various pulse sequences for fast proton spectroscopic imaging (SI) using the steady-state free precession (SSFP) condition are proposed. The sequences use either only the FID-like signal S1, only the echo-like signal S2, or both signals in separate but adjacent acquisition windows. As in SSFP imaging, S1 and S2 are separated by spoiler gradients. RF excitation is performed by slice-selective or chemical shift-selective pulses. The signals are detected in absence of a B0 gradient. Spatial localization is achieved by phase-encoding gradients which are applied prior to and rewound after each signal acquisition. Measurements with 2D or 3D spatial resolution were performed at 4.7 T on phantoms and healthy rat brain in vivo allowing the detection of uncoupled and J-coupled spins. The main advantages of SSFP based SI are the short minimum total measurement time (Tmin) and the high signal-to-noise ratio per unit measurement time (SNRt). The methods are of particular interest at higher magnetic field strength B0, as TR can be reduced with increasing B0 leading to a reduced Tmin and an increased SNRt. Drawbacks consist of the limited spectral resolution, particularly at lower B0, and the dependence of the signal intensities on T1 and T2. Further improvements are discussed including optimized data processing and signal detection under oscillating B0 gradients leading to a further reduction in Tmin. Magn Reson Med 50:453–460, 2003. © 2003 Wiley-Liss, Inc.

A new fast spectroscopic imaging (SI) method is presented which is based on spatial localization by the fast MRI method of rapid acquisition with relaxation enhancement (RARE) and encoding of the chemical shift information by shifting the position of a refocusing 180° pulse in a series of measurements. This method is termed spectroscopic RARE. In contrast to spectroscopic ultrafast low-angle RARE (U-FLARE), the formation of two echo families (odd and even) is suppressed by using a train of 180° RF pulses with an internal four-step phase cycle. By this means a high signal-to-noise ratio (SNR) per unit measurement time is obtained, because the separation of odd and even echoes, as well as dummy echoes to stabilize the echo amplitudes, is not needed anymore. The method is of particular interest for detecting signals of coupled spins, as effective homonuclear decoupling can be achieved by use of constant evolution time chemical shift encoding. The pulse sequence was implemented on a 4.7 T imaging system, tested on phantoms, and applied to the healthy rat brain in vivo. Spectroscopic RARE is particularly useful if T ≪ T2, which is typically fulfilled for in vivo proton SI measurements at high magnetic field strength. Magn Reson Med 47:523–528, 2002. © 2002 Wiley-Liss, Inc.

Proton magnetic resonance spectroscopy (1H-MRS) is a quantitative MR imaging technique often used to complement conventional MR imaging with specific metabolic information. A key metabolite is the amino acid derivative N-Acetylaspartate (NAA) which is an accepted marker to measure the extent of neurodegeneration in multiple sclerosis (MS) patients. NAA is catabolized by the enzyme aspartoacylase (ASPA) which is predominantly expressed in oligodendrocytes. Since the formation of MS lesions is paralleled by oligodendrocyte loss, NAA might accumulate in the brain, and therefore, the extent of neurodegeneration might be underestimated. In the present study, we used the well-characterized cuprizone model. There, the loss of oligodendrocytes is paralleled by a reduction in ASPA expression and activity as demonstrated by genome-wide gene expression analysis and enzymatic activity assays. Notably, brain levels of NAA were not increased as determined by gas chromatography–mass spectrometry and 1H-MRS. These important findings underpin the reliability of NAA quantification as a valid marker for the paraclinical determination of the extent of neurodegeneration, even under conditions of oligodendrocyte loss in which impaired metabolization of NAA is expected. Future studies have to reveal whether other enzymes are able to metabolize NAA or whether an excess of NAA is cleared by other mechanisms rather than enzymatic metabolism.

3D imaging has a significant impact on many challenges in life sciences, because biology is a 3-dimensional phenomenon. Current 3D imaging-technologies (various types MRI, PET, SPECT) are labeled, i.e. they trace the localization of a specific compound in the body. In contrast, 3D MALDI mass spectrometry-imaging (MALDI-MSI) is a label-free method imaging the spatial distribution of molecular compounds. It complements 3D imaging labeled methods, immunohistochemistry, and genetics-based methods. However, 3D MALDI-MSI cannot tap its full potential due to the lack of statistical methods for analysis and interpretation of large and complex 3D datasets. To overcome this, we established a complete and robust 3D MALDI-MSI pipeline combined with efficient computational data analysis methods for 3D edge preserving image denoising, 3D spatial segmentation as well as finding colocalized m/z values, which will be reviewed here in detail. Furthermore, we explain, why the integration and correlation of the MALDI imaging data with other imaging modalities allows to enhance the interpretation of the molecular data and provides visualization of molecular patterns that may otherwise not be apparent. Therefore, a 3D data acquisition workflow is described generating a set of 3 different dimensional images representing the same anatomies. First, an in-vitro MRI measurement is performed which results in a three-dimensional image modality representing the 3D structure of the measured object. After sectioning the 3D object into N consecutive slices, all N slices are scanned using an optical digital scanner, enabling for performing the MS measurements. Scanning the individual sections results into low-resolution images, which define the base coordinate system for the whole pipeline. The scanned images conclude the information from the spatial (MRI) and the mass spectrometric (MALDI-MSI) dimension and are used for the spatial three-dimensional reconstruction of the object performed by image registration techniques. Different strategies for automatic serial image registration applied to MS datasets are outlined in detail. The third image modality is histology driven, i.e. a digital scan of the histological stained slices in high-resolution. After fusion of reconstructed scan images and MRI the slice-related coordinates of the mass spectra can be propagated into 3D-space. After image registration of scan images and histological stained images, the anatomical information from histology is fused with the mass spectra from MALDI-MSI. As a result of the described pipeline we have a set of 3 dimensional images representing the same anatomies, i.e. the reconstructed slice scans, the spectral images as well as corresponding clustering results, and the acquired MRI. Great emphasis is put on the fact that the co-registered MRI providing anatomical details improves the interpretation of 3D MALDI images. The ability to relate mass spectrometry derived molecular information with in vivo and in vitro imaging has potentially important implications. This article is part of a Special Issue entitled: Computational Proteomics in the Post-Identification Era. Guest Editors: Martin Eisenacher and Christian Stephan.

Diffusion-weighted proton MR spectroscopy and imaging have been applied to a rat brain model of unilateral middle cerebral artery occlusion between 1 and 4 hr post occlusion. Similar apparent diffusion coefficients (ADC) of most metabolites were observed within each hemisphere. In the ischemic ipsilateral hemisphere, the ADCs were (0.083--0.116). 10(-3) mm(2)/sec for lactate (Lac), alanine (Ala), gamma-amino butyric acid (GABA), N-acetyl aspartate (NAA), glutamine (Gln), glutamate (Glu), total creatine (tCr), choline-containing compounds (Cho), and myo-inositol (Ins), in the contralateral hemisphere (0.138--0.158). 10(-3) mm(2)/sec for NAA, Glu, tCr, Cho, and Ins. Higher ADCs was determined for taurine (Tau) in the ipsilateral (0.144. 10(-3) mm(2)/sec) and contralateral (0.198. 10(-3) mm(2)/sec) hemisphere. In the ischemic hemisphere, a relative ADC decrease to 65--75% was observed for NAA, Glu, tCr, Cho, Ins and Tau, which was similar to the decrease of the water ADC (to 67%). The results suggest a common cause of the observed ADC changes and provide a broader experimental basis to evaluate theories of water and metabolite diffusion. Magn Reson Med 45:383-389, 2001.

A nuclear magnetic resonance study of the middle cerebral artery occlusion in the rat is presented. Experiments were performed on seven animals before and after occlusion, which occurred in situ. The emphasis in this study was on evaluating rapid proton spectroscopic imaging. Data were acquired with experimental durations of between 4 and 15 minutes for a 32 by 32 spatial matrix, with 64 spectroscopic data points per spatial element. The spectroscopic data were interleaved with diffusion-weighted nuclear magnetic resonance water images of the same slice. The study was terminated at about 6 hours after occlusion. The brains were then frozen in liquid nitrogen for biochemical imaging. The results showed that the signal from N-acetyl aspartate decreased and that of lactate increased within the infarcted region. The temporal course of these intensity changes varied between animals. Nineteen cortical spreading depressions (CSD) were observed by electrophysiologic monitoring during the experiments. Of these, 11 could be unambiguously detected in the lactate images, and a further 3 were on the threshold of detectability. As only a single slice could be examined, it is possible that the centers of depression for the remaining 6 CSD were outside the slice. To the authors' knowledge, this is the first report of the measurement of CSD using proton spectroscopic imaging. Thus, it is shown that this method is valuable not only in following the continuous evolution of proton metabolites with a good spatial and temporal resolution, but also in observing transient phenomena which are believed to play an important role in the expansion of the infarcted territory.

Tumorous and contralateral rat brain was examined by in vivo single voxel proton NMR spectroscopy. Magnetization transfer (MT) experiments cause attenuation of various metabolite signals. Selective saturation of immobile metabolites was achieved by pulsed RF preirradiation. The method is compared with continuous wave MT generation. In contralateral tissue, MT attenuation is detected for both the CH3 and the CH2 protons of (phospho-)creatine (Cr + PCr) and for a signal at 3.44 ppm ascribed to taurine. Significant attenuation is also observed for a signal at 3.78 ppm that is commonly ascribed to the alphaCH proton of glutamate and glutamine (Glx); however, no effect is observed for the gammaCH2 protons of Glx. Within implanted F98 glioma tumors, only the CH3 signal of Cr + PCr shows significant MT attenuation. Although the MT effect detected for lactate in the tumors fails to reach significance, a significant effect is observed for the lactate signal acquired during 3 to 9 min postmortem.

The standard model of magnetization transfer consists of six coupled, first-order differential equations which describe a lossless exchange of magnetization between two sites. The system of differential equations is solved semi-analytically in full generality. The solution allows one to model any experiment generating magnetization transfer. It is especially useful in investigation spin systems subjected to pulsed magnetization transfer experiments.

The decomposition of iron(III) acetylacetonate in high-boiling polyols such as diethylene glycole is an efficient way to produce water-soluble iron oxide nanoparticles (IONPs) with small sizes. We present an extension of this method by introducing ethylene diamine (EDA) or diethylene triamine (DTA) as a structure-directing agent and adding polyvinylpyrrolidone (PVP) as a stabilizing agent. The synthesis was studied with respect to effects of the chain length of the polyol used as solvent, the chain length of the structure-directing agent, the presence of PVP, the heating rate, and the nature of the precursor. By varying these parameters, we were able to show, that probably an interplay of the structure-directing agent and the polyol plays an important role for the stabilization and growth of the different facets of the IONP crystal. The chain length of the polyol used as solvent alters the influence of EDA or DTA as stabilizer of {1 1 1} facets, leading to IONPs with spherical, tetrahedral, or nanoplate morphology and mean diameters ranging from 4 nm up to 25 nm. PVP in the reaction medium narrows down particle size and shape distributions and promotes the formation of very stable, water-based colloidal solutions. The saturation magnetization of the particles was determined by a superconducting quantum interference device (SQUID) and their ability to act as a T2-contrast agent was tested by magnetic resonance imaging (MRI).

A k-space weighted spectroscopic imaging (SI) method is presented that allows a reduction in the total data acquisition time by up to 55% compared with standard SI. The k-space weighting is achieved by varying the repetition time, thus realizing an inherent apodization that corresponds to a circularly symmetric generalized Hamming filter. The flip angle is varied with the repetition time to enhance the signal-to-noise ratio. These techniques were employed using a short echo time of 10 ms. In vivo measurements on healthy rat brain at 4.7 T were conducted, obtaining two-dimensional spectroscopic imaging data from a 25 x 25 circularly reduced k-space area in as little as 5 min. The signal-to-noise ratio is sufficiently high to detect J-coupled resonances such as myo-inositol or glutamate/glutamine, demonstrating the ability to combine short acquisition times with comprehensive metabolic information. The T 1 dependency of the apodization and the corresponding point spread function was evaluated by computer simulations. The achievable signal-to-noise ratio per unit time was compared with standard SI giving a parameter-dependent advantage of approximately 20% of the standard SI method.

Intrastriatal injection of the excitotoxin N-methyl-dsc-aspartate (NMDA) in neonatal rat brain resulted in an acute ipsilateral decrease of the apparent diffusion coefficient (ADC) of brain tissue water, as measured with diffusion-weighted MRI. The early diffusion changes were accompanied by only mild changes in the overall metabolic status as measured by in vivo1H MRS and 31P MRS and metabolic imaging of brain sections. Minimal decreases in the high-energy phosphate levels and a small hemispheric acidosis were observed in the first 6¿h after NMDA administration. In addition, there was very modest lactate accumulation. Twenty-four hours after the induction of the excitotoxic injury the tissue energy status was still only moderately affected, whereas an overall decrease of 1H MRS-detected brain metabolites was found. Treatment with the non-competitive NMDA-antagonist MK-801 given within 90 min after NMDA injection rapidly reversed the NMDA-induced changes in the entire ipsilateral hemisphere. The effect of the competitive NMDA-antagonist D-CPPene was restricted to the cortical areas and was accomplished on a slower time scale. Our results indicate that; (i) early excitotoxicity in the neonatal rat brain does not lead to profound changes in the metabolic status; and (ii) brain tissue water ADC changes are not necessarily associated with a metabolic energy failure.

The potential of two-dimensional (2D)-J NMR for in vivo proton MRS is examined. Single voxel measurements on the rat brain were performed at 4.7 T using point-resolved spectrocopy localization with a voxel size of 64 microliter and total measuring times of 10-15 min. It is shown that a series of measurements with only 16 or fewer different echo times (TE) enables good signal localization in the f1 axis corresponding to the coupling patterns. For data evaluation, the 2D-J NMR spectrum as well as cross-sections at given f1 values and projections onto the f2 axis are used. A comparison between cross-section spectra taken at different f1 values may help to solve problems of peak assignment. The projection of the 2D magnitude spectrum onto the f2 axis corresponds to a homonuclear decoupled 1D proton spectrum. Because the T2 relaxation times of several coupled resonances (e.g., myo-inositol and glutamate) are rather long, only minor losses in the quality of the projection spectra occur if the measurements with short TE (< or = 50 ms) are not used for data processing. Thus, homonuclear decoupled proton spectra detecting uncoupled and several coupled resonances can be measured with high quality in vivo, even on MR systems that are not equipped with actively shielded gradients, prohibiting data acquisition with TEs of 50 ms or less.

Simultaneous multislice proton spectroscopic imaging (SI) is presented using a pulse sequence with multifrequency selective RF excitation and Hadamard encoding in the slice direction, and conventional Fourier phase encoding in the in-plane directions. Double-echo data acquisition is used to increase the spectral information of the experiment. Tests on a phantom demonstrate the quality of the slice selection. Results of in vivo measurements of the healthy rat brain show that spectra with a high signal-to-noise ratio can be acquired from four slices within 32 min. The measurements were performed at 4.7 T using a field of view of 32 x 32 mm2, a slice thickness of 3 mm, and a voxel size of 12 microliters. The proposed method is a useful alternative to sequential multislice SI and 3D SI. Furthermore, it is possible to combine sequential and simultaneous multislice SI.

Isotopic fingerprinting was evaluated for its potential to generate characteristic fingerprints of crop protection products in an extensive survey, using the insecticide Fipronil. One hundred and twenty batches of Fipronil from the BASF production site in France were analyzed for the isotope ratios of δ(13)C, δ(15)N, and δ(34)S. Samples spanned a production time of four years and were analyzed by elemental analysis, coupled to isotope ratio mass spectrometry (EA/IRMS). A number of Fipronil samples from other sources were analyzed in the same manner and were compared to the samples from BASF by means of multivariate data analysis. The isotopic fingerprint was sufficiently specific to differentiate between Fipronil from BASF production and Fipronil from other producers. This suggests that isotopic fingerprinting is suitable for the authenticity control of active compounds in crop protection products. It is anticipated that this technique will deliver great benefit in the defense against counterfeits and illegal parallel imports.