Nuclear Overhauser Enhancement Research Articles

Magnetization transfer weighted laminar fMRI with multi-echo FLASH

Laminar functional magnetic resonance imaging (fMRI) using the gradient echo (GRE) blood oxygenation level dependent (BOLD) contrast is prone to signal changes arising from large unspecific venous vessels. Alternatives based on changes of cerebral blood volume (CBV) become more popular since it is expected that this hemodynamic response is dominant in microvasculature. One approach to sensitize the signal toward changes in CBV, and to simultaneously reduce unwanted extravascular (EV) BOLD blurring, is to selectively reduce gray matter (GM) signal via magnetization transfer (MT). In this work, we use off-resonant MT-pulses with a 3D FLASH readout to perform MT-prepared (MT-prep) laminar fMRI of the primary visual cortex (V1) at multiple echo times at 7 T. With a GRE-BOLD contrast without additional MT-weighting as reference, we investigated the influence of the MT-preparation on the shape and the echo time dependency of laminar profiles. Through numerical simulations, we optimized the sequence parameters to increase the sensitivity toward signal changes induced by changes in arterial CBV and to delineate the contributions of different compartments to the signal. We show that at 7 T, GM signals can be reduced by 30 %. Our laminar fMRI responses exhibit an increased signal change in the parenchyma at very short TE compared to a BOLD-only reference as a result of reduced EV signal intensity. By varying echo times, we could show that MT-prep results in less sensitivity toward unwanted signal changes based on changes in T2*. We conclude that when accounting for nuclear overhauser enhancement effects in blood, off-resonant MT-prep combined with efficient short TE readouts can become a promising method to reduce unwanted EV venous contributions in GRE-BOLD and/or to allow scanning at much shorter echo times without incurring a sensitivity penalty in laminar fMRI.

Reassembled saturation transfer (REST) MR images at 2 B1 values for in vivo exchange-dependent imaging of amide and nuclear Overhauser enhancement.

Design an efficient CEST scheme for exchange-dependent images with high contrast-to-noise ratio. Reassembled saturation transfer (REST) signals were defined as r.Z=r.Zref - r.ZCEST and the reassembled exchange-dependen magnetization transfer ratio r.MTRRex =r.1/Zref - r.1/ZCEST , utilizing the averages over loosely sampled reference frequency offsets as Zref and over densely sampled target offsets as ZCEST . Using r.MTRRex measured under 2 B1,sat values, exchange rate could be estimated. The REST approach was optimized and assessed quantitatively by simulations for various exchange rates, pool concentration, and water T1 . In vivo evaluation was performed on ischemic rat brains at 7Tesla and human brains at 3Tesla, in comparison with conventional asymmetrical analysis, Lorentzian difference (LD), an MTRRex_ LD. For a broad choice of ranges and numbers, Δr.Z and r.MTRRex exhibited comparable quantification features with conventional LD and MTRRex _LD, respectively, when B1,sat ≤ 1μT. The subtraction of 2 REST values under distinct B1,sat values showed linear relationships with exchange rate and obtained immunity to field inhomogeneity and variation in MT and water T1 . For both rat and human studies, REST images exhibited similar contrast distribution to MTRRex _LD, with superiority in contrast-to-noise ratio and acquisition efficiency. Compared with MTRRex _LD, 2-B1,sat subtraction REST images displayed better resistance to B1 inhomogeneity, with more specific enhanced regions. They also showed higher signals for amide than for nuclear Overhauser enhancement effect in human brain, presumably reflecting the higher increment from faster-exchanging species as B1,sat increased. Featuring high contrast-to-noise ratio efficiency, REST could be a practical exchange-dependent approach readily applicable to either retrospective Z-spectra analysis or perspective 6-offset acquisition.

Noninvasively differentiating acute and chronic nephropathies via multiparametric urea-CEST, nuclear Overhauser enhancement-CEST, and quantitative magnetization transfer MRI.

Standardized blood tests often lack adequate sensitivity and specificity to capture the gradual progression of renal injuries. We suggest a multiparametric molecular MRI approach as a noninvasive tool for monitoring renal function loss and distinguishing different types of renal injuries. CEST and quantitative magnetization transfer (qMT) imaging were performed on cisplatin (n=16) and aristolochic acid (AA)-induced nephropathy (n=22) mouse models at 7T with an infusion of either saline or urea. Seven-pool Lorentzian fitting was applied for the analysis of CEST Z-spectra, and the T1 -corrected CEST contrast apparent exchange-dependent relaxation (AREX) from urea (+1ppm) and two nuclear Overhauser enhancement (NOE) pools (-1.6 and -3.5ppm) were measured. Similarly, qMT spectra were fitted into two-pool Ramani equation and the relative semi-solid macromolecular pool-size ratio was measured. Histology of mouse kidneys was performed to validate the MR findings. AA model showed disrupted spatial gradients of urea in the kidney and significantly decreased NOE CEST and qMT contrast. The cisplatin model showed slightly decreased qMT contrast only. The orrelation of MR parameters to histological features showed that NOE CEST and qMT imaging are sensitive to both acute and chronic injuries, whereas urea CEST shows a significant correlation only to acute injuries. These results indicate that our multiparametric approach allows comprehensive and totally noninvasive monitoring of renal function and histological changes for distinguishing different nephropathies.

NOE-weighted imaging in tumors using low-duty-cycle 2π-CEST.

Nuclear Overhauser enhancement (NOE)-mediated CEST imaging at -3.5 ppm has shown clinical interest in diagnosing tumors. Multiple-pool Lorentzian fit has been used to quantify NOE, which, however, requires a long scan time. Asymmetric analysis of CEST signals could be a simple and fast method to quantify this NOE, but it has contamination from the amide proton transfer (APT) at 3.5 ppm. This work proposes a new method using an asymmetric analysis of a low-duty-cycle pulsed-CEST sequence with a flip angle of 360°, termed 2π-CEST, to reduce the contribution from APT. Simulations were used to evaluate the capability of the 2π-CEST to reduce APT. Experiments on animal tumor models were performed to show its advantages compared with the conventional asymmetric analysis. Samples of reconstituted phospholipids and proteins were used to evaluate the molecular origin of this NOE. The 2π-CEST has reduced contribution from APT. In tumors where we show that the NOE is comparable to the APT effect, reducing the contamination from APT is crucial. The results show that the NOE signal obtained with 2π-CEST in tumor regions appears more homogeneous than that obtained with the conventional method. The phantom study showed that both phospholipids and proteins contribute to the NOE at -3.5 ppm. The NOE at -3.5 ppm has a different contrast mechanism from APT and other CEST/NOE effects. The proposed 2π-CEST is more accurate than the conventional asymmetric analysis in detecting NOE, and requires much less scan time than the multiple-pool Lorentzian fit.

Advances in the exact nuclear Overhauser effect 2018-2022.

The introduction of the exact nuclear Overhauser enhancement (eNOE) methodology to solution-state nuclear magnetic resonance (NMR) spectroscopy results in tighter distance restraints from NOEs than in convention analysis. These improved restraints allow for higher resolution in structure calculation and even the disentanglement of different conformations of macromolecules. While initial work primarily focused on technical development of the eNOE, structural studies aimed at the elucidation of spatial sampling in proteins and nucleic acids were published in parallel prior to 2018. The period of 2018-2022 saw a continued series of technical innovation, but also major applications addressing biological questions. Here, we review both aspects, covering topics from the implementation of non-uniform sampling of NOESY buildups, novel pulse sequences, adaption of the eNOE to solid-state NMR, advances in eNOE data analysis, and innovations in structural ensemble calculation, to applications to protein, RNA, and DNA structure elucidation.

Proton-detected solution-state NMR at 14.1 T based on scalar-driven 13C Overhauser dynamic nuclear polarization

Overhauser dynamic nuclear polarization (ODNP) NMR of solutions at high fields is usually mediated by scalar couplings that polarize the nuclei of heavier, electron-rich atoms. This leaves 1H-detected NMR outside the realm of such studies. This study presents experiments that deliver 1H-detected NMR experiments on relatively large liquid volumes (60 ∼ 100 μL) and at high fields (14.1 T), while relying on ODNP enhancements. To this end 13C NMR polarizations were first enhanced by relying on a mechanism that utilizes e--13C scalar coupling interactions; the nuclear spin alignment thus achieved was then passed on to neighboring 1H for observation, by a reverse INEPT scheme relying on one-bond JCH-couplings. Such 13C →1H polarization transfer ported the 13C ODNP gains into the 1H, permitting detection at higher frequencies and with higher potential sensitivities. For a model solution of labeled 13CHCl3 comixed with a nitroxide-based TEMPO derivative as polarizing agent, an ODNP enhancement factor of ca. 5x could thus be imparted to the 1H signal. When applied to bigger organic molecules like 2-13C-phenylacetylene and 13C8-indole, ODNP enhancements in the 1.2-3x range were obtained. Thus, although handicapped by the lower γ of the 13C, enhancements could be imparted on the 1H thermal acquisitions in all cases. We also find that conventional 1H–13C nuclear Overhauser enhancements (NOEs) are largely absent in these solutions due to the presence of co-dissolved radicals, adding negligible gains and playing negligible roles on the scalar e-→13C ODNP transfer. Potential rationalizations of these effects as well as extensions of these experiments, are briefly discussed.

Synergism between betaine surfactants under high-salt conditions

Foam properties and surface dilational rheology of single and mixed LHSB (lauramidopropyl hydroxyl sulfobetaine)-OAB (oleic amide propyl betaine) surfactants were measured to investigate the synergism between the two components. Experimental results showed obvious synergistic effects in foam stability, dilational modulus, and phase angle between LHSB and OAB. Moreover, interaction parameters were calculated to quantitatively characterize the synergism. The negative interaction parameters between LHSB and OAB demonstrated that there was an attractive interaction between LHSB and OAB, and the attractive interaction contributed to forming the dense surfactant layer at the liquid/air layer, which in turn improved foam stability. The negative values of parameters reached their maximum when the LHSB: OAB molar ratio ranged from 1:1 to 1:2, the same time the foam stability reached its maximum. The interaction between LHSB and OAB was confirmed by 2D NOSEY (two-dimensional nuclear overhauser enhancement spectroscopy) experiments, the interaction between LHSB and OAB could be attributed to the hydrogen bonding or the electrostatic interaction between the hydrophilic groups of the two surfactants.

In vivo pH mapping with omega plot-based quantitative chemical exchange saturation transfer MRI.

Chemical exchange saturation transfer (CEST) MRI is promising for detecting dilute metabolites and microenvironment properties, which has been increasingly adopted in imaging disorders such as acute stroke and cancer. However, in vivo CEST MRI quantification remains challenging because routine asymmetry analysis (MTRasym ) or Lorentzian decoupling measures a combined effect of the labile proton concentration and its exchange rate. Therefore, our study aimed to quantify amide proton concentration and exchange rate independently in a cardiac arrest-induced global ischemia rat model. The amide proton CEST (APT) effect was decoupled from tissue water, macromolecular magnetization transfer, nuclear Overhauser enhancement, guanidinium, and amine protons using the image downsampling expedited adaptive least-squares (IDEAL) fitting algorithm on Z-spectra obtained under multiple RF saturation power levels, before and after global ischemia. Omega plot analysis was applied to determine amide proton concentration and exchange rate simultaneously. Global ischemia induces a significant APT signal drop from intact tissue. Using the modified omega plot analysis, we found that the amide proton exchange rate decreased from 29.6 ± 5.6 to 12.1 ± 1.3s-1 (P < 0.001), whereas the amide proton concentration showed little change (0.241 ± 0.035% vs. 0.202 ± 0.034%, P=0.074) following global ischemia. Our study determined the labile proton concentration and exchange rate underlying the in vivo APT MRI. The significant change in the exchange rate, but not the concentration of amide proton demonstrated that the pH effect dominates the APT contrast during tissue ischemia.

NMR Observation of Sulfhydryl Signals in SARS-CoV-2 Main Protease Aids Structural Studies.

The 68‐kDa homodimeric 3C‐like protease of SARS‐CoV‐2, Mpro (3CLpro/Nsp5), is a key antiviral drug target. NMR spectroscopy of this large system proved challenging and resonance assignments have remained incomplete. Here we present the near‐complete (>97 %) backbone assignments of a C145A variant of Mpro (Mpro C145A) both with, and without, the N‐terminal auto‐cleavage substrate sequence, in its native homodimeric state. We also present SILLY (Selective Inversion of thioL and Ligand for NOESY), a simple yet effective pseudo‐3D NMR experiment that utilizes NOEs to identify interactions between Cys‐thiol or aliphatic protons, and their spatially proximate backbone amides in a perdeuterated protein background. High protection against hydrogen exchange is observed for 10 of the 11 thiol groups in Mpro C145A, even those that are partially accessible to solvent. A combination of SILLY methods and high‐resolution triple‐resonance NMR experiments reveals site‐specific interactions between Mpro, its substrate peptides, and other ligands, which present opportunities for competitive binding studies in future drug design efforts.

Quasi-steady-state amide proton transfer (QUASS APT) MRI enhances pH-weighted imaging of acute stroke.

Chemical exchange saturation transfer (CEST) imaging measurement depends not only on the labile proton concentration and pH-dependent exchange rate but also on experimental conditions, including the relaxation delay and radiofrequency (RF) saturation time. Our study aimed to extend a quasi-steady-state (QUASS) solution to a modified multi-slice CEST MRI sequence and test if it provides enhanced pH imaging after acute stroke. Our study derived the QUASS solution for a modified multislice CEST MRI sequence with an unevenly segmented RF saturation between image readout and signal averaging. Numerical simulation was performed to test if the generalized QUASS solution corrects the impact of insufficiently long relaxation delay, primary and secondary saturation times, and multi-slice readout. In addition, multiparametric MRI scans were obtained after middle cerebral artery occlusion, including relaxation and CEST Z-spectrum, to evaluate the performance of QUASS CEST MRI in a rodent acute stroke model. We also performed Lorentzian fitting to isolate multi-pool CEST contributions. The QUASS analysis enhanced pH-weighted magnetization transfer asymmetry contrast over the routine apparent CEST measurements in both contralateral normal (-3.46% ± 0.62% (apparent) vs. -3.67% ± 0.66% (QUASS), P&lt; 0.05) and ischemic tissue (-5.53% ± 0.68% (apparent) vs. -5.94% ± 0.73% (QUASS), P&lt; 0.05). Lorentzian fitting also showed significant differences between routine and QUASS analysis of ischemia-induced changes in magnetization transfer, amide, amine, guanidyl CEST, and nuclear Overhauser enhancement (-1.6parts per million) effects. Our study demonstrated that generalized QUASS analysis enhanced pH MRI contrast and improved quantification of the underlying CEST contrast mechanism, promising for further in vivo applications.

An improved, time-efficient approach to extract accurate distance restraints for NMR2 structure calculation.

Exact nuclear Overhauser enhancement (eNOE) yields highly accurate, ensemble averaged H-H distance restraints with an accuracy of up to 0.1 Å for the multi-state structure determination of proteins as well as for nuclear magnetic resonance molecular replacement (MR) to determine the structure of the protein-ligand interaction site in a time-efficient manner. However, in the latter application, the acquired eNOEs lack the obtainable precision of 0.1 Å because of the asymmetrical nature of the filtered nuclear Overhauser enhancement spectroscopy (NOESY) experiment used in MR. This error is further propagated to the eNOE equations used to fit and extract the distance restraints. In this work, a new analysis method is proposed to obtain inter-molecular distance restraints from the filtered NOESY spectrum more accurately and intuitively by dividing the NOE cross peak by the corresponding diagonal peak of the ligand. The method termed diagonal-normalised eNOEs was tested on the data acquired by on the complex of PIN1 and a small, weak-binding phenylimidazole fragment. MR calculations performed using the distances derived from diagonal-normalised eNOEs yielded the right orientation of the fragment in the binding pocket and produced a structure that more closely resembles the benchmark X-ray structure (2XP6) with an average heavy-atom root-mean-square deviation (RMSD) of 1.681 Å with respect to it, when compared to the one produced with traditional MR with an average heavy atom RMSD of 3.628 Å. This is attributed to the higher precision of the evaluated distance restraints.

Breast cancer imaging with glucosamine CEST (chemical exchange saturation transfer) MRI: first human experience.

This study aims to evaluate the feasibility of imaging breast cancer with glucosamine (GlcN) chemical exchange saturation transfer (CEST) MRI technique to distinguish between tumor and surrounding tissue, compared to the conventional MRI method. Twelve patients with newly diagnosed breast tumors (median age, 53 years) were recruited in this prospective IRB-approved study, between August 2019 and March 2020. Informed consent was obtained from all patients. All MRI measurements were performed on a 3-T clinical MRI scanner. For CEST imaging, a fat-suppressed 3D RF-spoiled gradient echo sequence with saturation pulse train was applied. CEST signals were quantified in the tumor and in the surrounding tissue based on magnetization transfer ratio asymmetry (MTRasym) and a multi-Gaussian fitting. GlcN CEST MRI revealed higher signal intensities in the tumor tissue compared to the surrounding breast tissue (MTRasym effect of 8.12 ± 4.09%, N = 12, p = 2.2 E-03) with the incremental increase due to GlcN uptake of 3.41 ± 0.79% (N = 12, p = 2.2 E-03), which is in line with tumor location as demonstrated by T1W and T2W MRI. GlcN CEST spectra comprise distinct peaks corresponding to proton exchange between free water and hydroxyl and amide/amine groups, and relayed nuclear Overhauser enhancement (NOE) from aliphatic groups, all yielded larger CEST integrals in the tumor tissue after GlcN uptake by an averaged factor of 2.2 ± 1.2 (p = 3.38 E-03), 1.4 ± 0.4 (p =9.88 E-03), and 1.6 ± 0.6 (p = 2.09 E-02), respectively. The results of this initial feasibility study indicate the potential of GlcN CEST MRI to diagnose breast cancer in a clinical setup. • GlcN CEST MRI method is demonstrated for its the ability to differentiate between breast tumor lesions and the surrounding tissue, based on the differential accumulation of the GlcN in the tumors. • GlcN CEST imaging may be used to identify metabolic active malignant breast tumors without using a Gd contrast agent. • The GlcN CEST MRI method may be considered for use in a clinical setup for breast cancer detection and should be tested as a complementary method to conventional clinical MRI methods.

Demonstration of fast and equilibrium human muscle creatine CEST imaging at 3 T.

Creatine chemical exchange saturation transfer (CrCEST) MRI is used increasingly in muscle imaging. However, the CrCEST measurement depends on the RF saturation duration (Ts) and relaxation delay (Td), and it is challenging to compare the results of different scan parameters. Therefore, this study aims to evaluate the quasi-steady-state (QUASS) CrCEST MRI on clinical 3T scanners. T1 and CEST MRI scans of Ts/Td of 1 s/1 s and 2 s/2 s were obtained from a multi-compartment creatine phantom and 5 healthy volunteers. The CrCEST effect was quantified with asymmetry analysis in the phantom, whereas 5-pool Lorentzian fitting was applied to isolate creatine from phosphocreatine, amide proton transfer, combined magnetization transfer and nuclear Overhauser enhancement effects, and direct water saturation in four major muscle groups of the lower leg. The routine and QUASS CrCEST measurements were compared under two different imaging conditions. Paired Student's t-test was performed with p-values less than 0.05 considered statistically significant. The phantom study showed a substantial influence of Ts/Td on the routine CrCEST quantification (p= 0.02), and such impact was mitigated with the QUASS algorithm (p= 0.20). The volunteer experiment showed that the routine CrCEST, amide proton transfer, and combined magnetization transfer and nuclear Overhauser enhancement effects increased significantly with Ts and Td (p< 0.05) and were significantly smaller than the corresponding QUASS indices (p< 0.01). In comparison, the QUASS CrCEST MRI showed little dependence on Ts and Td, indicating its robustness and accuracy. The QUASS CrCEST MRI is feasible to provide fast and accurate muscle creatine imaging.

Molecular dynamics simulation or structure refinement of proteins: are solvent molecules required? A case study using hen lysozyme

In protein simulation or structure refinement based on values of observable quantities measured in (aqueous) solution, solvent (water) molecules may be explicitly treated, omitted, or represented by a potential of mean-solvation-force term, depending on protein coordinates only, in the force field used. These three approaches are compared for hen egg white lysozyme (HEWL). This 129-residue non-spherical protein contains a variety of secondary-structure elements, and ample experimental data are available: 1630 atom–atom Nuclear Overhauser Enhancement (NOE) upper distance bounds, 213 3 J-couplings and 200 S2 order parameters. These data are used to compare the performance of the three approaches. It is found that a molecular dynamics (MD) simulation in explicit water approximates the experimental data much better than stochastic dynamics (SD) simulation in vacuo without or with a solvent-accessible-surface-area (SASA) implicit-solvation term added to the force field. This is due to the missing energetic and entropic contributions and hydrogen-bonding capacities of the water molecules and the missing dielectric screening effect of this high-permittivity solvent. Omission of explicit water molecules leads to compaction of the protein, an increased internal strain, distortion of exposed loop and turn regions and excessive intra-protein hydrogen bonding. As a consequence, the conformation and dynamics of groups on the surface of the protein, which may play a key role in protein–protein interactions or ligand or substrate binding, may be incorrectly modelled. It is thus recommended to include water molecules explicitly in structure refinement of proteins in aqueous solution based on nuclear magnetic resonance (NMR) or other experimentally measured data.

Incorporating NOE-Derived Distances in Conformer Generation of Cyclic Peptides with Distance Geometry.

Nuclear magnetic resonance (NMR) data from NOESY (nuclear Overhauser enhancement spectroscopy) and ROESY (rotating frame Overhauser enhancement spectroscopy) experiments can easily be combined with distance geometry (DG) based conformer generators by modifying the molecular distance bounds matrix. In this work, we extend the modern DG based conformer generator ETKDG, which has been shown to reproduce experimental crystal structures from small molecules to large macrocycles well, to include NOE-derived interproton distances. In noeETKDG, the experimentally derived interproton distances are incorporated into the distance bounds matrix as loose upper (or lower) bounds to generate large conformer sets. Various subselection techniques can subsequently be applied to yield a conformer bundle that best reproduces the NOE data. The approach is benchmarked using a set of 24 (mostly) cyclic peptides for which NOE-derived distances as well as reference solution structures obtained by other software are available. With respect to other packages currently available, the advantages of noeETKDG are its speed and that no prior force-field parametrization is required, which is especially useful for peptides with unnatural amino acids. The resulting conformer bundles can be further processed with the use of structural refinement techniques to improve the modeling of the intramolecular nonbonded interactions. The noeETKDG code is released as a fully open-source software package available at www.github.com/rinikerlab/customETKDG.

3D APT and NOE CEST-MRI of healthy volunteers and patients with non-enhancing glioma at 3\xa0T

ObjectiveClinical application of chemical exchange saturation transfer (CEST) can be performed with investigation of amide proton transfer (APT) and nuclear Overhauser enhancement (NOE) effects. Here, we investigated APT- and NOE-weighted imaging based on advanced CEST metrics to map tumor heterogeneity of non-enhancing glioma at 3 T.Materials and methodsAPT- and NOE-weighted maps based on Lorentzian difference (LD) and inverse magnetization transfer ratio (MTRREX) were acquired with a 3D snapshot CEST acquisition at 3 T. Saturation power was investigated first by varying B1 (0.5–2 µT) in 5 healthy volunteers then by applying B1 of 0.5 and 1.5 µT in 10 patients with non-enhancing glioma. Tissue contrast (TC) and contrast-to-noise ratios (CNR) were calculated between glioma and normal appearing white matter (NAWM) and grey matter, in APT- and NOE-weighted images. Volume percentages of the tumor showing hypo/hyperintensity (VPhypo/hyper,CEST) in APT/NOE-weighted images were calculated for each patient.ResultsLD APT resulting from using a B1 of 1.5 µT was found to provide significant positive TCtumor,NAWM and MTRREX NOE (B1 of 1.5 µT) provided significant negative TCtumor,NAWM in tissue differentiation. MTRREX-based NOE imaging under 1.5 µT provided significantly larger VPhypo,CEST than MTRREX APT under 1.5 µT.ConclusionThis work showed that with a rapid CEST acquisition using a B1 saturation power of 1.5 µT and covering the whole tumor, analysis of both LD APT and MTRREX NOE allows for observing tumor heterogeneity, which will be beneficial in future studies using CEST-MRI to improve imaging diagnostics for non-enhancing glioma.

Chemical exchange saturation transfer imaging of creatine, phosphocreatine, and protein arginine residue in tissues.

Chemical exchange saturation transfer (CEST) MRI has become a promising technique to assay target proteins and metabolites through their exchangeable protons, noninvasively. The ubiquity of creatine (Cr) and phosphocreatine (PCr) due to their pivotal roles in energy homeostasis through the creatine phosphate pathway has made them prime targets for CEST in the diagnosis and monitoring of disease pathologies, particularly in tissues heavily dependent on the maintenance of rich energy reserves. Guanidinium CEST from protein arginine residues (i.e. arginine CEST) can also provide information about the protein profile in tissue. However, numerous obfuscating factors stand as obstacles to the specificity of arginine, Cr, and PCr imaging through CEST, such as semisolid magnetization transfer, fast chemical exchanges such as primary amines, and the effects of nuclear Overhauser enhancement from aromatic and amide protons. In this review, the specific exchange properties of protein arginine residues, Cr, and PCr, along with their validation, are discussed, including the considerations necessary to target and tune their signal effects through CEST imaging. Additionally, strategies that have been employed to enhance the specificity of these exchanges in CEST imaging are described, along with how they have opened up possible applications of protein arginine residues, Cr and PCr CEST imaging in the study and diagnosis of pathology. A clear understanding of the capabilities and caveats of using CEST to image these vital metabolites and mitigation strategies is crucial to expanding the possibilities of this promising technology.

Accelerating quantitative 13C NMR spectra using an EXtended ACquisition Time (EXACT) method.

Accurate quantitative 13C NMR spectra can be accelerated by using EXACT (EXtended ACquisition Time) NMR methods which reduce Nuclear Overhauser Enhancement (NOE) during the FID. This allows 30-50% shorter experiment times to be used when achieving a given level of quantitative accuracy.

Fast and equilibrium CEST imaging of brain tumor patients at 3T

Chemical exchange saturation transfer (CEST) MRI, versatile for detecting endogenous mobile proteins and tissue pH, has proved valuable in tumor imaging. However, CEST MRI scans are often performed under non-equilibrium conditions, which confound tissue characterization. This study proposed a quasi-steady-state (QUASS) CEST MRI algorithm to standardize fast and accurate tumor imaging at 3 T. The CEST signal evolution was modeled by longitudinal relaxation rate during relaxation delay (Td) and spinlock relaxation during RF saturation time (Ts), from which the QUASS CEST effect is derived. Numerical simulation and human MR imaging experiments (7 healthy volunteers and 19 tumor patients) were conducted at 3 T to compare the CEST measurements obtained under two representative experimental conditions. In addition, amide proton transfer (APT), combined magnetization transfer (MT) and nuclear overhauser enhancement (NOE) effects, and direct water saturation were isolated using a 3-pool Lorentzian fitting in white matter and gray matter of healthy volunteers and for patients in the contralateral normal-appearing white matter and tumor regions. Finally, the student's t-test was performed between conventional and QUASS CEST measurements. The routine APT and combined MT & NOE measures significantly varied with Ts and Td (P < .001) and were significantly smaller than the corresponding QUASS indices (P < .001). In contrast, the results from the QUASS reconstruction showed little dependence on the scan protocol (P > .05), indicating the accuracy and robustness of QUASS CEST MRI for tumor imaging. To summarize, the QUASS CEST reconstruction algorithm enables fast and accurate tumor CEST imaging at 3 T, promising to expedite and standardize clinical CEST MRI.

Sulfa drugs–based Norbornenyl imides and reductive Heck reactions: Synthesis and antimicrobial screening

AbstractIn the field of our research interest, we designed new sulfa drugs–substituted norbornyl imides as prospective bioactive scaffolds by the reaction of endo‐endic anhydride with sulfa drugs followed by reductive Heck reactions of these products. Norbornenyl imides, as starting compounds, and their Heck products were achieved in good yields. Molecular characterization and stereochemistry of the compounds were investigated using Fourier transform infrared, liquid chromatography time‐of‐flight mass spectrometry, proton nuclear magnetic resonance, carbon 13 attached proton test, H‐H correlation spevtroscopy, and nuclear overhauser enhancement spectroscopy experiments. The Heck products of norbornenyl imides showed exo‐selectivity. All eight novel synthesized compounds, including two sulfa drugs–based norbornenyl imides and six exo‐5‐aryl‐ substituted norbornyl imides, as well as standard sulfonamides, were evaluated for antimicrobial activity against nine microorganisms consisting of Gram‐negative bacteria, Gram‐positive nonactinobacteria, actinobacteria, and yeast. The most active compound was 3a against actinobacteria. The efficacy of 2 and its derivatives against the studied strains were weaker than 3 and its derivatives, but all the studied samples exhibited antifungal potency.