Single 4D Research Articles

Regional Patterns of Pulsatility Index and Wall Shear Stress Across Cerebral Circulation of Adolescents with High Insulin Resistance

BackgroundInsulin resistance (IR) is associated with altered arterial stiffness, blood flow, and shear stress patterns in the peripheral circulation of middle‐aged and older diabetic and non‐diabetic hypertensive populations. Both altered blood flow and shear stress patterns may contribute to the elevated risk of cerebrovascular disease in IR populations. The aim of this study was to determine if adolescents (12–18yrs) with high IR (indexed as High Homeostatic Model Assessment of IR, HOMA‐IR) display early signs of cerebrovascular dysfunction. We hypothesized that young adolescents with a high HOMA‐IR would exhibit increased stiffness and decreased shear patterns in cerebral conduit arteries.Methods2 healthy controls (17±3) and 3 IR subjects (14±1) underwent a single 4D Flow MRI scan (scan time ~ 5min, no contrast given) to quantify cerebral blood flow (CBF), pulsatility index (PI) and wall shear stress (WSS) in extracranial and intracranial conduit arteries. 11 arteries were analyzed: middle cerebral (MCA) x2, anterior cerebral (ACA) x2, posterior cerebral (PCA) x2, vertebral (VA) x2, internal carotid (ICA) x2, and basilar. PI was calculated for each artery as [(CBFmax − CBFmin)/CBFmean]. WSS was measured by segmenting the vascular tree from PC angiograms and calculating the velocity gradient at the vessel wall using 4D flow MRI data over ~3 voxel. Due to voxel limitations, the PCA value is exploratory. Total CBF was calculated as the sum of both ICA and both VA. Significance was determined using a two‐tail Welch's t‐test.ResultsIndividuals with IR displayed higher HOMA‐IR (IR 5.3±0.2, Ctrl, 3.3±0.3, p=0.02), BMI (kg•m2; IR 28±2, Ctrl 19±4, p=0.03) and fasting insulin (μU/mL; IR 25±4, Ctrl 13±0, p=0.04), but fasting glucose was not different (mg/dL; IR 79±4, Ctrl 93±5, p=0.07). PI was not different between groups among the 11 conduit arteries. However, there was a trend for higher PI in the IR group in the left PCA and right VA. Resting WSS did not differ between Ctrl and IR among the 11 conduit arteries. Similarly, total resting CBF was not different between Ctrl and IR (mL/min; IR 811±115, Ctrl 735±79, p=0.64).ConclusionPI or WSS were similar in 11 conduit arteries of cerebral circulation in adolescent controls compared to IR. These data indicate IR does not cause measurable arterial stiffening or altered total cerebral blood flow patterns. Therefore, the pathogenic arterial characteristics observed in middle‐aged adults with IR may be linked to the duration and/or severity of IR in association with other known vascular pathological processes linked to aging.Support or Funding InformationThis research was made possible by the National Science Foundation Grant #HRD‐1500138 (UW‐Madison WiscAMP‐BD program) and UW Graduate School Pilot Funding.

Facilitating unambiguous NMR assignments and enabling higher probe density through selective labeling of all methyl containing amino acids.

The deuteration of proteins and selective labeling of side chain methyl groups has greatly enhanced the molecular weight range of proteins and protein complexes which can be studied using solution NMR spectroscopy. Protocols for the selective labeling of all six methyl group containing amino acids individually are available, however to date, only a maximum of five amino acids have been labeled simultaneously. Here, we describe a new methodology for the simultaneous, selective labeling of all six methyl containing amino acids using the 115kDa homohexameric enzyme CoaD from E. coli as a model system. The utility of the labeling protocol is demonstrated by efficiently and unambiguously assigning all methyl groups in the enzymatic active site using a single 4D (13)C-resolved HMQC-NOESY-HMQC experiment, in conjunction with a crystal structure. Furthermore, the six fold labeled protein was employed to characterize the interaction between the substrate analogue (R)-pantetheine and CoaD by chemical shift perturbations, demonstrating the benefit of the increased probe density.

A REVIEW OF MULTI-HAZARD RISK ASSESSMENT (MHRA) USING 4D DYNAMIC MODELS

Abstract. This paper reviews the 4D dynamic models for multi natural hazard risk assessment. It is important to review the characteristic of the different dynamic models and to choose the most suitable model for certain application. The characteristic of the different 4D dynamic models are based on several main aspects (e.g. space, time, event or phenomenon etc.). The most suitable 4D dynamic model depends on the type of application it is used for. There is no single 4D Dynamic model suitable for all types of application. Therefore, it is very important to define the requirements of the 4D Dynamic model. The main context of this paper is spatio temporal modelling for multi hazards.

Modeling a 3D City Model and Its Levels of Detail as a True 4D Model

The various levels of detail (LODs) of a 3D city model are often stored independently, without links between the representations of the same object, causing inconsistencies, as well as update and maintenance problems. One solution to this problem is to model the LOD as an extra geometric dimension perpendicular to the three spatial ones, resulting in a true 4D model in which a single 4D object (a polychoron) represents a 3D polyhedral object (e.g., a building) at all of its LODs and a multiple-LOD 3D city model is modeled as a 4D cell complex. While such an approach has been discussed before at a conceptual level, our objective in this paper is to describe how it can be realized by appropriately linking existing 3D models of the same object at different LODs. We first present our general methodology to construct such a 4D model, which consists of three steps: (1) finding corresponding 0D–3D cells; (2) creating 1D–4D cells connecting them; and (3) constructing the 4D model. Because of the complex relationships between the objects in different LODs, the creation of the connecting cells can become difficult. We therefore describe four different alternatives to do this, and we discuss the advantages and disadvantages of each in terms of their feasibility in practice and the properties that the resulting 4D model has. We show how the different linking schemes result in objects with different characteristics in several use cases. We also show how our linking method works in practice by implementing the linking of matching cells to construct a 4D model.

Technical prerequisites and imaging protocols for CT perfusion imaging in oncology

The aim of this review article is to define the technical prerequisites of modern state-of-the-art CT perfusion imaging in oncology at reasonable dose levels. The focus is mainly on abdominal and thoracic tumor imaging, as they pose the largest challenges with respect to attenuation and patient motion. We will show that low kV dynamic scanning in conjunction with detection technology optimized for low photon fluxes has the highest impact on reducing dose independently of other choices made in the protocol selection. We discuss, derived from relatively simple first principles, on what appropriate temporal sampling and total scan duration depend on and why optimized contrast medium injection protocols are also essential in limiting dose. Finally we will examine the possibility of simultaneously extracting standard morphological and functional information from one single 4D examination as a potential enabler for a more widespread use of dynamic contrast enhanced CT in oncology.

Respiratory-induced venous blood flow effects using flexible retrospective double-gating.

To demonstrate a novel velocity sensitive acquisition and retrospective cardiorespiratory double-gated reconstruction scheme to examine respiratory effect on venous blood flow in healthy volunteers. Radial two dimensional (2D) phase contrast MR is performed at 3 Tesla in the internal jugular vein (IJV) of healthy volunteers (n = 6). Data are retrospectively partitioned based on respiratory waveforms using three schemes: moving average for respiration plateaus, gradient for active respiration, and ten respiratory phases that are cardiac time-averaged. A single 4D flow MR scan is performed in the neck of a healthy volunteer. After gradient operation, blood velocity measurements are made along the IJV length. Percent changes from expiration to inspiration for moving average and gradient techniques are statistically compared with paired t-tests. Percent change increase in summed IJV mean and peak blood flow during active inspiration versus active expiration in 2D was significant (mean flow: 11.5 ± 8.0%, peak flow: 11.9 ± 5.9%, P < 0.01). Smallest cross-sectional area and largest blood velocity are seen during inspiration phases (phase number: area-6.5 ± 3.6, velocity-6.2 ± 3.2). Significant increase in mean velocity along the length of the IJV was observed in 3D, with increasing percent changes more proximal to the chest (mean, 39 ± 30%; range, 0-93%, P = 0.001). With a radial acquisition, this pilot study demonstrates feasibility of simultaneous retrospective cardiorespiratory gating in IJV flow. Greatest differences in flow occur between active respiration phases, increasing in magnitude more proximal to the chest.

Non-invasive assessment of cardiac function and pulmonary vascular resistance in an canine model of acute thromboembolic pulmonary hypertension using 4D flow cardiovascular magnetic resonance.

BackgroundThe purpose of this study was to quantify right (RV) and left (LV) ventricular function, pulmonary artery flow (QP), tricuspid valve regurgitation velocity (TRV), and aorta flow (QS) from a single 4D flow cardiovascular magnetic resonance (CMR) (time-resolved three-directionally motion encoded CMR) sequence in a canine model of acute thromboembolic pulmonary hypertension (PH).MethodsAcute PH was induced in six female beagles by microbead injection into the right atrium. Pulmonary arterial (PAP) and pulmonary capillary wedge (PCWP) pressures and cardiac output (CO) were measured by right heart catheterization (RHC) at baseline and following induction of acute PH. Pulmonary vascular resistance (PVRRHC) was calculated from RHC values of PAP, PCWP and CO (PVRRHC = (PAP-PCWP)/CO). Cardiac magnetic resonance (CMR) was performed on a 3 T scanner at baseline and following induction of acute PH. RV and LV end-diastolic (EDV) and end-systolic (ESV) volumes were determined from both CINE balanced steady-state free precession (bSSFP) and 4D flow CMR magnitude images. QP, TRV, and QS were determined from manually placed cutplanes in the 4D flow CMR flow-sensitive images in the main (MPA), right (RPA), and left (LPA) pulmonary arteries, the tricuspid valve (TRV), and aorta respectively. MPA, RPA, and LPA flow was also measured using two-dimensional flow-sensitive (2D flow) CMR.ResultsBiases between 4D flow CMR and bSSFP were 0.8 mL and 1.6 mL for RV EDV and RV ESV, respectively, and 0.8 mL and 4 mL for LV EDV and LV ESV, respectively. Flow in the MPA, RPA, and LPA did not change after induction of acute PAH (p = 0.42-0.81). MPA, RPA, and LPA flow determined with 4D flow CMR was significantly lower than with 2D flow (p < 0.05). The correlation between QP/TRV and PVRRHC was 0.95. The average QP/QS was 0.96 ± 0.11.ConclusionsUsing both magnitude and flow-sensitive data from a single 4D flow CMR acquisition permits simultaneous quantification of cardiac function and cardiopulmonary hemodynamic parameters important in the assessment of PH.

Vacuum Referred Binding Energy of the Single 3d, 4d, or 5d Electron in Transition Metal and Lanthanide Impurities in Compounds

The vacuum referred binding energy (VRBE) of the single electron in the lowest energy 3d level of Sc2 +, V4 +, Cr5 +, the lowest 4d level of Y2 +, Zr3 +, Nb4 +, Mo5 + and the lowest 5d level of Ta4 +, and W5 + in various compounds are determined by means of the chemical shift model. They will be compared with the VRBE in the already established lowest 3d level of Ti3 + and the lowest 5d level of Eu2 + and Ce3 +. Clear trends with changing charge of the transition metal (TM) cation and with changing principle quantum number n = 3, 4, or 5 of the nd level will be identified. This work will demonstrate that the trends correlate with the VRBE in the free ion nd TM cation level. The acquired knowledge on the VRBE of the electron in the nd TM impurity levels but also on TM based compounds with nd type of conduction band bottom provides new insight in the luminescence properties of TM activated compounds.

Fermion actions extracted from lattice super Yang-Mills theories

We revisit 2D $\mathcal{N}=(2,2)$ super Yang-Mills lattice formulation (Sugino model) to investigate its fermion action with two (Majorana) fermion flavors and exact chiral-$U(1)_{R}$ symmetry. We show that the reconcilement of chiral symmetry and absence of further species-doubling originates in the 4D clifford algebra structure of the action, where 2D two flavors are spuriously treated as a single 4D four-spinor with four 4D gamma matrices introduced into kinetic and Wilson terms. This fermion construction based on the higher-dimensional clifford algebra is extended to four dimensions in two manners: (1) pseudo-8D sixteen-spinor treatment of 4D four flavors with eight 8D gamma matrices, (2) pseudo-6D eight-spinor treatment of 4D two flavors with five out of six 6D gamma matrices. We obtain 4D four-species and two-species lattice fermions with unbroken subgroup of chiral symmetry and other essential properties. We discuss their relations to staggered and Wilson twisted-mass fermions. We also discuss their potential feedback to 4D super Yang-Mills lattice formulations.

Measuring Pulsatile Flow in Cerebral Arteries Using 4D Phase-Contrast MR Imaging

4D PCMRI can be used to quantify pulsatile hemodynamics in multiple cerebral arteries. The aim of this study was to compare 4D PCMRI and 2D PCMRI for assessments of pulsatile hemodynamics in major cerebral arteries. We scanned the internal carotid artery, the anterior cerebral artery, the basilar artery, and the middle cerebral artery in 10 subjects with a single 4D and multiple 2D PCMRI acquisitions by use of a 3T system and a 32-channel head coil. We assessed the agreement regarding net flow and the volume of arterial pulsatility (ΔV) for all vessels. 2D and 4D PCMRI produced highly correlated results, with r = 0.86 and r = 0.95 for ΔV and net flow, respectively (n = 69 vessels). These values increased to r = 0.93 and r = 0.97, respectively, during investigation of a subset of measurements with <5% variation in heart rate between the 4D and 2D acquisition (n = 31 vessels). Significant differences were found for ICA and MCA net flow (P = .004 and P < .001, respectively) and MCA ΔV (P = .006). However, these differences were attenuated and no longer significant when the subset with stable heart rate (n = 31 vessels) was analyzed. 4D PCMRI provides a powerful methodology to measure pulsatility of the larger cerebral arteries from a single acquisition. A large part of differences between measurements was attributed to physiologic variations. The results were consistent with 2D PCMRI.

Four‐dimensional phase contrast MRI With accelerated dual velocity encoding

AbstractPurpose:To validate a novel approach for accelerated four‐dimensional phase contrast MR imaging (4D PC‐MRI) with an extended range of velocity sensitivity.Materials and Methods:4D PC‐MRI data were acquired with a radially undersampled trajectory (PC‐VIPR). A dual Venc (dVenc) processing algorithm was implemented to investigate the potential for scan time savings while providing an improved velocity‐to‐noise ratio. Flow and velocity measurements were compared with a flow pump, conventional 2D PC MR, and single Venc 4D PC‐MRI in the chest of 10 volunteers.Results:Phantom measurements showed excellent agreement between accelerated dVenc 4D PC‐MRI and the pump flow rate (R2 ≥ 0.97) with a three‐fold increase in measured velocity‐to‐noise ratio (VNR) and a 5% increase in scan time. In volunteers, reasonable agreement was found when combining 100% of data acquired with Venc = 80 cm/s and 25% of the high Venc data, providing the VNR of a 80 cm/s acquisition with a wider velocity range of 160 cm/s at the expense of a 25% longer scan.Conclusion:Accelerated dual Venc 4D PC‐MRI was demonstrated in vitro and in vivo. This acquisition scheme is well suited for vascular territories with wide ranges of flow velocities such as congenital heart disease, the hepatic vasculature, and others. J. Magn. Reson. Imaging 2012. © 2012 Wiley Periodicals, Inc.

Localized substance delivery to single cell and 4D imaging of its uptake using a flow channel with a lateral aperture

We have developed a novel microfluidic device for space- and time-resolved (4D) visualization of intracellular events when a cell surface is partially exposed to external stimuli. The device, fabricated using 3D rotational inclined UV lithography of photoresist SU-8, consists of a cell-containing chamber and a flow channel separated by a thin vertical wall having a lateral micrometer aperture smaller than a cell. A cell is first immobilized on the aperture by suction from the flow channel using a syringe pump, and a chemical stimulant is then fed to the channel so that only the cell surface bounded by the aperture is subjected to the stimulus without leakage to other part of the cell surface. The subsequent lateral signal propagation inside the cell can be visualized using high-speed fluorescence confocal microscopy. As an experimental demonstration of the device, 2-NBDG (fluorescence glucose analog) intake into a mouse insulinoma cell, MIN6m9, was visualized in 4D resolution.

Abnormalities in resting-state functional connectivity in early human immunodeficiency virus infection.

Limited information is available concerning changes that occur in the brain early in human immunodeficiency virus (HIV) infection. This investigation evaluated resting-state functional connectivity, which is based on correlations of spontaneous blood oxygen level-dependent functional magnetic resonance imaging (fMRI) oscillations between brain regions, in 15 subjects within the first year of HIV infection and in 15 age-matched controls. Resting-state fMRI data for each session were concatenated in time across subjects to create a single 4D dataset and decomposed into 36 independent component analysis (ICA) using Multivariate Exploratory Linear Optimized Decomposition into Independent Components. ICA components were back-reconstructed for each subject's 4D data to estimate subject-specific spatial maps using the dual-regression technique. Comparison of spatial maps between HIV and controls revealed significant differences in the lateral occipital cortex (LOC) network. Reduced coactivation in left inferior parietal cortex within the LOC network was identified in the HIV subjects. Connectivity strength within this region correlated with performance on tasks involving visual-motor coordination (Grooved Pegboard and Rey Figure Copy) in the HIV group. The findings indicate prominent changes in resting-state functional connectivity of visual networks early in HIV infection. This network may sustain injury in association with the intense viremia and brain viral invasion before immune defenses can contain viral replication. Resting-state functional connectivity may have utility as a noninvasive neuroimaging biomarker for central nervous system impairment in early HIV infection.

A non-uniformly sampled 4D HCC(CO)NH-TOCSY experiment processed using maximum entropy for rapid protein sidechain assignment

One of the stiffest challenges in structural studies of proteins using NMR is the assignment of sidechain resonances. Typically, a panel of lengthy 3D experiments are acquired in order to establish connectivities and resolve ambiguities due to overlap. We demonstrate that these experiments can be replaced by a single 4D experiment that is time-efficient, yields excellent resolution, and captures unique carbon–proton connectivity information. The approach is made practical by the use of non-uniform sampling in the three indirect time dimensions and maximum entropy reconstruction of the corresponding 3D frequency spectrum. This 4D method will facilitate automated resonance assignment procedures and it should be particularly beneficial for increasing throughput in NMR-based structural genomics initiatives.

Holographic dual of QCD from black D5 branes

We study the dynamics of probe D7 flavor probe branes in the background of near extremal D5 branes. This model is a holographic dual to a gauge theory with spontaneous breaking of a U(Nf)L × U(Nf)R chiral symmetry. The spectrum of two such D7 embeddings, contains a single massive 4D meson coming from the world volume U(1) gauge field, the pion, and a single massive 4D scalar meson coming from fluctuations of the embedding of the brane. In addition, there are continuum five dimensional states due to the finite height of the effective potential in the radial direction. We investigate baryons in this model, and find that the size is stabilized due to the Chern-Simons term in the D7 world volume action. The model admits a Hagedorn temperature of 1/2πR where R is the radius parameter in the D5 branes metric. We investigate the pattern of chiral symmetry breaking in the deconfined phase as a function of the asymptotic separation of the branes L. We find that for πR/3 < L 1.068R that chiral symmetry is restored, and that chiral symmetry is broken for L outside this window. We further argue that the solutions with L < π/3 are only classically stable, and in fact no D7 embedding exists with these boundary conditions.

SU‐GG‐T‐460: Multiple Gating for Lung Stereotactic Body Radiotherapy Treatment

Purpose: An inherent problem in treatment of lung cancers with stereotactic body radiotherapy (SBRT) is target motion. Multiple‐phase‐based gating refers to treating the target at two or more phase windows with plans optimized for each window individually. It closely approximates the four‐dimensional (4D) tracking technique when large number of gating windows is used. Compared to conventional single gating window technique, multiple‐phase‐based gating can improve the normal tissue sparing by taking advantage of the location change of the target. In addition, the duty cycle may be further improved. Compared to 4D tracking, the dose conformity of the multiple‐gating technique may be inferior. However, its implementation may be more robust because of the allowance of residual error inside each gating window. In this study, we evaluate the dose performances for different techniques including static, single gated, dynamic 4D tracking, and two‐window‐gated SBRT plans. Method and Materials: A motion phantom was scanned with 4DCT and planned using the above techniques. The motion was 1D sinusoidal with amplitude of 1cm. For any optimized plan, 95% of the PTV was covered by the prescription dose of 60Gy. Film dosimetry was performed to compare the doses delivered in each plans. The above techniques were also compared for a lung patient who was undergoing SBRT treatment, prescribed to 60Gy in 3 fractions. Results: Both the phantom and the patient studies showed similar results. If normalized to the static plan, the V20's were decreased by 56%,22%,16%, and the maximum doses in “lung” were changed to 102%,58%,45%, for single window gating, two‐window gating, and 4D tracking, respectively. Conclusion: Multiple‐gating technique has significant improvement in lung dose sparing compared with static or single‐gating techniques. Its difference from 4D plan is relatively small. The dose performance also highly depends on the size of tumor and the extent of target motion.

Scanner characterization for color measurement and diagnostics

We propose a novel scanner characterization approach for applications requiring color measurement of hardcopy output in calibration, characterization, and diagnostics applications. The method is advantageous for common practical color printing sys- tems that use more than the minimum of three colorants necessary for subtractive color reproduction; printing with cyan (C), magenta (M), yellow (Y), and black (K) is the most prevalent example we use in our description. The proposed method exploits the fact that for the scenarios in consideration, in addition to the scanner RGB values for a scanned patch, the CMYK control values used to print the patch are also available and can be exploited in characterization. An indexed family of 3D scanner characterizations is created, each characterization providing a mapping from scanner RGB to CIELAB for a fixed value of K, the latter constituting the index for the char- acterization. Combined together, the family of 3D characterizations provides a single 4D characterization that maps scanner RGB ob- tained from scanning a patch and the K control value used for print- ing the patch to a colorimetric CIELAB measurement for the patch. A significant improvement in the robustness of the method to varia- tions in printing is obtained by modifying the K index to utilize the scanned output for a black-only patch printed with the correspond- ing K value instead of directly utilizing the control K value used at the printer. Results show that the proposed 4D scanner character- ization technique can significantly outperform standard 3D ap- proaches in the target applications. © 2007 SPIE and IS&T.

Four-dimensional heteronuclear correlation experiments for chemical shift assignment of solid proteins

Chemical shift assignment is the first step in all established protocols for structure determination of uniformly labeled proteins by NMR. The explosive growth in recent years of magic-angle spinning (MAS) solid-state NMR (SSNMR) applications is largely attributable to improved methods for backbone and side-chain chemical shift correlation spectroscopy. However, the techniques developed so far have been applied primarily to proteins in the size range of 5-10 kDa, despite the fact that SSNMR has no inherent molecular weight limits. Rather, the degeneracy inherent to many 2D and 3D SSNMR spectra of larger proteins has prevented complete unambiguous chemical shift assignment. Here we demonstrate the implementation of 4D backbone chemical shift correlation experiments for assignment of solid proteins. The experiments greatly reduce spectral degeneracy at a modest cost in sensitivity, which is accurately described by theory. We consider several possible implementations and investigate the CANCOCX pulse sequence in detail. This experiment involves three cross polarization steps, from H to CA[i], CA[i] to N[i], and N[i] to C'[i-1], followed by a final homonuclear mixing period. With short homonuclear mixing times (<20 ms), backbone correlations are observed with high sensitivity; with longer mixing times (>200 ms), long-range correlations are revealed. For example, a single 4D experiment with 225 ms homonuclear mixing time reveals approximately 200 uniquely resolved medium and long-range correlations in the 56-residue protein GB1. In addition to experimental demonstrations in the 56-residue protein GB1, we present a theoretical analysis of anticipated improvements in resolution for much larger proteins and compare these results in detail with the experiments, finding good agreement between experiment and theory under conditions of stable instrumental performance.

Characterization of full‐length hepatitis C virus genotype 4 sequences

Over 85% of the world's nearly 170 million hepatitis C virus (HCV)-infected subjects exist in regions of Africa, Southeast Asia and Middle Eastern countries where genotypes 4-6 are very common. In particular, HCV genotype 4 is highly prevalent in Egypt with more than 19% of the population infected and chronic HCV representing one of the top five leading causes of death, due in part to ineffective interferon alpha treatment against this genotype. Despite this, very little work has been carried out to characterize the sequence diversity of genotype 4, which will be critical to the development of effective vaccines and antiviral therapies against this genotype. As a result of the paucity of sequence data available for HCV genotype 4, for which only one full genome sequence is currently available, we were interested in characterizing additional genotype 4 sequences and to provide reagents for amplification of this genotype. Here we describe seven unique HCV genotype 4a full genomes, in addition to a single genotype 4d genome, and characterize their sequence diversity in relation to other more closely characterized HCV genotypes.

Determination of all NOes in 1H–13C–Me-ILV-U−2H–15N Proteins with Two Time-Shared Experiments

We present two time-shared experiments that enable the characterization of all nOes in 1H-13C-ILV methyl-labelled proteins that are otherwise uniformly deuterated and 15N enriched and possibly selectively protonated for distinct residue types. A 3D experiment simultaneously provides the spectra of a 3D NOESY-HN-TROSY and of a 3D NOESY-HC-PEP-HSQC. Thus, nOes from any protons to methyl or amide protons are dispersed with respect to 15N and 13C chemical shifts, respectively. The single 4D experiment presented here yields simultaneously the four 4D experiments HC-HSQC-NOESY-HC-PEP-HSQC, HC-HSQC-NOESY-HN-TROSY, HN-HSQC-NOESY-HN-TROSY and HN-HSQC-NOESY-HC-PEP-HSQC. This allows for the unambiguous determination of all nOes involving amide and methyl protons. The method was applied to a (1H,13C)-ILV-(1H)-FY-(U-2H,15N) sample of a 37 kDa di-domain of the E. coli enterobactin synthetase module EntF.