Spin Labeling Method Research Articles

Oxygen extraction fraction mapping based combining quantitative susceptibility mapping and quantitative blood oxygenation level-dependent imaging model using multi-delay PCASL

Background and purposeThe oxygen extraction fraction is an essential biomarker for the assessment of brain metabolism. A recently proposed method combined with quantitative susceptibility mapping and quantitative blood oxygen level-dependent magnitude enables noninvasive mapping of the oxygen extraction fraction. Our study investigated the oxygen extraction fraction mapping variations of single-delay and multi-delay arterial spin-labeling. Materials and methodsA total of twenty healthy participants were enrolled. The multi-echo spoiled gradient-echo, multi-delay arterial spin-labeling, and magnetization-prepared rapid gradient echo sequences were acquired at 3.0 T. The mean oxygen extraction fraction was generated under a single delay time of 1780 ms, multi-delay arterial spin-labeling of transit-corrected cerebral blood flow, and multi-delay arterial spin-labeling of arterial cerebral blood volume. The results were compared via paired t tests and the Wilcoxon test. Linear regression analyses were used to investigate the relationships among the oxygen extraction fraction, cerebral blood flow, and venous cerebral blood volume. ResultsThe oxygen extraction fraction estimate with multi-delay arterial spin-labeling yielded a significantly lower value than that with single-delay arterial spin-labeling. The average values for the whole brain under single-delay arterial spin-labeling, multi-delay arterial spin-labeling of transit-corrected cerebral blood flow, and multi-delay arterial spin-labeling of arterial cerebral blood volume were 41.5 ± 1.7 % (P < 0.05), 41.3 ± 1.9 % (P < 0.001), and 40.9 ± 1.9 % (N = 20), respectively. The oxygen extraction fraction also showed a significant inverse correlation with the venous cerebral blood volume under steady-state conditions when multi-delay arterial spin-labeling was used (r = 0.5834, p = 0.0069). ConclusionThese findings suggest that the oxygen extraction fraction is significantly impacted by the arterial spin-labeling methods used in the quantitative susceptibility mapping plus the quantitative blood oxygen level-dependent model, indicating that the differences should be accounted for when employing oxygen extraction fraction mapping based on this model in diseases.

Association of Alpha-Crystallin with Human Cortical and Nuclear Lens Lipid Membrane Increases with the Grade of Cortical and Nuclear Cataract.

Eye lens α-crystallin has been shown to become increasingly membrane-bound with age and cataract formation; however, to our knowledge, no studies have investigated the membrane interactions of α-crystallin throughout the development of cataracts in separated cortical membrane (CM) and nuclear membrane (NM) from single human lenses. In this study, four pairs of human lenses from age-matched male and female donors and one pair of male lenses ranging in age from 64 to 73 years old (yo) were obtained to investigate the interactions of α-crystallin with the NM and CM throughout the progression of cortical cataract (CC) and nuclear cataract (NC) using the electron paramagnetic resonance spin-labeling method. Donor health history information (diabetes, smoker, hypertension, radiation treatment), sex, and race were included in the data analysis. The right eye lenses CM and NM investigated were 64 yo male (CC: 0), 68 yo male (CC: 3, NC: 2), 73 yo male (CC: 1, NC: 2), 68 yo female (CC: 3, NC: 2), and 73 yo female (CC: 1, NC: 3). Similarly, left eye lenses CM and NM investigated were 64 yo male (CC: 0), 68 yo male (CC: 3, NC: 2), 73 yo male (CC: 2, NC: 3), 68 yo female (CC: 3, NC: 2), and 73 yo female (CC: 1, NC: 3). Analysis of α-crystallin binding to male and female eye lens CM and NM revealed that the percentage of membrane surface occupied (MSO) by α-crystallin increases with increasing grade of CC and NC. The binding of α-crystallin resulted in decreased mobility, increased order, and increased hydrophobicity on the membrane surface in male and female eye lens CM and NM. CM mobility decreased with an increase in cataracts for both males and females, whereas the male lens NM mobility showed no significant change, while female lens NM showed increased mobility with an increase in cataract grade. Our data shows that a 68 yo female donor (long-term smoker, pre-diabetic, and hypertension; grade 3 CC) showed the largest MSO by α-crystallin in CM from both the left and right lens and had the most pronounced mobility changes relative to all other analyzed samples. The variation in cholesterol (Chol) content, size and amount of cholesterol bilayer domains (CBDs), and lipid composition in the CM and NM with age and cataract might result in a variation of membrane surface mobility, membrane surface hydrophobicity, and the interactions of α-crystallin at the surface of each CM and NM. These findings provide insight into the effect of decreased Chol content and the reduced size and amount of CBDs in the cataractous CM and NM with an increased binding of α-crystallin with increased CC and NC grade, which suggests that Chol and CBDs might be a key component in maintaining lens transparency.

Cholesterol Content Regulates the Interaction of αA-, αB-, and α-Crystallin with the Model of Human Lens-Lipid Membranes.

α-Crystallin (αABc) is a major protein comprised of αA-crystallin (αAc) and αB-crystallin (αBc) that is found in the human eye lens and works as a molecular chaperone by preventing the aggregation of proteins and providing tolerance to stress. However, with age and cataract formation, the concentration of αABc in the eye lens cytoplasm decreases, with a corresponding increase in the membrane-bound αABc. This study uses the electron paramagnetic resonance (EPR) spin-labeling method to investigate the role of cholesterol (Chol) and Chol bilayer domains (CBDs) in the binding of αAc, αBc, and αABc to the Chol/model of human lens-lipid (Chol/MHLL) membranes. The maximum percentage of membrane surface occupied (MMSO) by αAc, αBc, and αABc to Chol/MHLL membranes at a mixing ratio of 0 followed the trends: MMSO (αAc) > MMSO (αBc) ≈ MMSO (αABc), indicating that a higher amount of αAc binds to these membranes compared to αBc and αABc. However, with an increase in the Chol concentration in the Chol/MHLL membranes, the MMSO by αAc, αBc, and αABc decreases until it is completely diminished at a mixing ratio of 1.5. The Ka of αAc, αBc, and αABc to Chol/MHLL membranes at a mixing ratio of 0 followed the trend: Ka (αBc) ≈ Ka (αABc) > Ka (αAc), but it was close to zero with the diminished binding at a Chol/MHLL mixing ratio of 1.5. The mobility near the membrane headgroup regions decreased with αAc, αBc, and αABc binding, and the Chol antagonized the capacity of the αAc, αBc, and αABc to decrease mobility near the headgroup regions. No significant change in membrane order near the headgroup regions was observed, with an increase in αAc, αBc, and αABc concentrations. Our results show that αAc, αBc, and αABc bind differently with Chol/MHLL membranes at mixing ratios of 0 and 0.5, decreasing the mobility and increasing hydrophobicity near the membrane headgroup region, likely forming the hydrophobic barrier for the passage of polar and ionic molecules, including antioxidants (glutathione), creating an oxidative environment inside the lens, leading to the development of cataracts. However, all binding was completely diminished at a mixing ratio of 1.5, indicating that high Chol and CBDs inhibit the binding of αAc, αBc, and αABc to membranes, preventing the formation of hydrophobic barriers and likely protecting against cataract formation.

Aberrant Neurovascular Coupling in Diabetic Retinopathy Using Arterial Spin Labeling (ASL) and Functional Magnetic Resonance Imaging (fMRI) methods.

Previous imaging studies have demonstrated that diabetic retinopathy (DR) is linked to structural and functional abnormalities in the brain. However, the extent to which DR patients exhibit abnormal neurovascular coupling remains largely unknown. Thirty-one patients with DR and 31 sex- and age-matched healthy controls underwent resting-state functional magnetic resonance imaging (rs-fMRI) to calculate functional connectivity strength (FCS) and arterial spin-labeling imaging (ASL) to calculate cerebral blood flow (CBF). The study compared CBF-FCS coupling across the entire grey matter and CBF/FCS ratios (representing blood supply per unit of connectivity strength) per voxel between the two groups. Additionally, a support vector machine (SVM) method was employed to differentiate between diabetic retinopathy (DR) patients and healthy controls (HC). In DRpatients compared to healthy controls, there was a reduction in CBF-FCS coupling across the entire grey matter. Specifically, DR patients exhibited elevated CBF/FCS ratios primarily in the primary visual cortex, including the right calcarine fissure and surrounding cortex. On the other hand, reduced CBF/FCS ratios were mainly observed in premotor and supplementary motor areas, including the left middle frontal gyrus. An elevated CBF/FCS ratio suggests that patients with DR may have a reduced volume of gray matter in the brain. A decrease in its ratio indicates a decrease in regional CBF in patients with DR. These findings suggest that neurovascular decoupling in the visual cortex, as well as in the supplementary motor and frontal gyrus, may represent a neuropathological mechanism in diabetic retinopathy.

A Protocol to Depict the Proteolytic Processes Using a Combination of Metal-Organic Materials (MOMs), Electron Paramagnetic Resonance (EPR), and Mass Spectrometry (MS).

Proteolysis is a critical biochemical process yet a challenging field to study experimentally due to the self-degradation of a protease and the complex, dynamic degradation steps of a substrate. Mass spectrometry (MS) is the traditional way for proteolytic studies, yet it is challenging when time-resolved, step-by-step details of the degradation process are needed. We recently found a way to resolve the cleavage site, preference/selectivity of cleavage regions, and proteolytic kinetics by combining site-directed spin labeling (SDSL) of protein substrate, time-resolved two-dimensional (2D) electron paramagnetic resonance (EPR) spectroscopy, protease immobilization via metal-organic materials (MOMs), and MS. The method has been demonstrated on a model substrate and protease, yet there is a lack of details on the practical operations to carry out our strategy. Thus, this protocol summarizes the key steps and considerations when carrying out the EPR/MS study on proteolytic processes, which can be generalized to study other protein/polypeptide substrates in proteolysis. Details for the experimental operation and cautions of each step are reported with figures illustrating the concepts. This protocol provides an effective approach to understanding the proteolytic process with the advantages of offering time-resolved, residue-level resolution of structural basis underlying the process. Such information is important for revealing the cleavage site and proteolytic mechanisms of unknown proteases. The advantage of EPR, probing the target substrate regardless of the complexities caused by the proteases and their self-degradation, offers a practically effective, rapid, and easy-to-operate approach to studying proteolysis. Key features • Combining protease immobilization, EPR, spin labeling, and MS experimental methods allows for the analysis of proteolysis process in real time. • Reveals cleavage site, kinetics of product generation, and preference of cleavage regions via time-resolved SDSL-EPR. • MS confirms EPR findings and helps depict the sequences and populations of the cleaved segments in real time. • The demonstrated method can be generalized to other proteins or polypeptide substrates upon proteolysis by other proteases.

The cortical high-flow sign of oligodendroglioma, IDH-mutant and 1p/19q-codeleted: comparison between arterial spin labeling and dynamic susceptibility contrast methods.

The cortical high-flow sign with the non-enhancing area was reportedly found to be more frequent with oligodendroglioma, IDH-mutant and 1p/19q codeleted (ODG IDHm-codel) than with IDH-wildtype or astrocytoma, IDH-mutant on arterial spin labeling (ASL) in diffuse gliomas. This study aimed to compare the identification rate of the cortical high-flow sign on ASL in patients with ODG IDHm-codel to that on dynamic susceptibility contrast-enhanced perfusion-weighted imaging (DSC-PWI). Participants consisted of 32 adult ODG IDHm-codel patients with pathologically confirmed. Subtraction images were generated from paired control and label images on ASL. For DSC, dynamic T2*-weighted perfusion weighted images were obtained after pre-bolus of gadolinium-based contrast agent. Regional cerebral blood flow/volume maps were generated based on the concentration-time curve and arterial input function. Tumor-affecting cortices without contrast enhancement on conventional MR imaging were targeted. The identification rate of the cortical high-flow sign was compared between ASL and DSC using the Pearson's Chi-Square test. Frequency of the cortical high-flow sign was significantly higher on ASL (18/32, 56.3%; p < 0.001) than on DSC (5/32, 15.6%). All cases with the positive cortical high-flow sign on DSC were identified on ASL. ASL effectively identifies the cortical high-flow sign in ODG IDHm-codel, surpassing DSC in identification rates.

Binding of βL-Crystallin with Models of Animal and Human Eye Lens-Lipid Membrane.

Several discoveries show that with age and cataract formation, β-crystallin binds with the lens membrane or associates with other lens proteins, which bind with the fiber cell plasma membrane, accompanied by light scattering and cataract formation. However, how lipids (phospholipids and sphingolipids) and cholesterol (Chol) influence β-crystallin binding to the membrane is unclear. This research aims to elucidate the role of lipids and Chol in the binding of β-crystallin to the membrane and the membrane's physical properties (mobility, order, and hydrophobicity) with β-crystallin binding. We used electron paramagnetic resonance (EPR) spin-labeling methods to investigate the binding of βL-crystallin with a model of porcine lens-lipid (MPLL), model of mouse lens-lipid (MMLL), and model of human lens-lipid (MHLL) membrane with and without Chol. Our results show that βL-crystallin binds with all of the investigated membranes in a saturation manner, and the maximum parentage of the membrane surface occupied (MMSO) by βL-crystallin and the binding affinity (Ka) of βL-crystallin to the membranes followed trends: MMSO (MPLL) > MMSO (MMLL) > MMSO (MHLL) and Ka (MHLL) > Ka (MMLL) ≈ Ka (MPLL), respectively, in which the presence of Chol reduces the MMSO and Ka for all membranes. The mobility near the headgroup regions of the membranes decreases with an increase in the binding of βL-crystallin; however, the decrease is more pronounced in the MPLL and MMLL membranes than the MHLL membrane. In the MPLL and MMLL membranes, the membranes become slightly ordered near the headgroup with an increase in βL-crystallin binding compared to the MHLL membrane. The hydrophobicity near the headgroup region of the membrane increases with βL-crystallin binding; however, the increase is more pronounced in the MPLL and MMLL membranes than the MHLL membrane, indicating that βL-crystallin binding creates a hydrophobic barrier for the passage of polar molecules, which supports the barrier hypothesis in cataract formation. However, in the presence of Chol, there is no significant increase in hydrophobicity with βL-crystallin binding, suggesting that Chol prevents the formation of a hydrophobic barrier, possibly protecting against cataract formation.

Quantification of Age-Related Changes in the Lateral Organization of the Lipid Portion of the Intact Membranes Isolated from the Left and Right Eye Lenses of the Same Human Donor.

The continuous wave EPR spin-labeling method was used to evaluate age-related changes in the amounts of phospholipids (PLs) and cholesterol (Chol) in domains present in intact, cortical, and nuclear fiber cell plasma membranes isolated separately from the left and right eye lenses of the same human donor. The relative amounts of boundary plus trapped PLs were evaluated with the PL analog 12-doxylstearic acid spin label (12-SASL) and the relative amounts of trapped Chol with the Chol analog androstane spin label (ASL). The donors ranged in age from 15 to 70 years. Both the left and right eye lenses from donors aged 60, 65, and 70 years had nuclear cataracts; additionally, the right eye lens only of the 60-year-old donor had a cortical cataract. In transparent lenses, the relative amounts of boundary plus trapped PLs increase monotonously with donor age, and, at all ages, this amount was greater in nuclear compared with cortical membranes. Moreover, in transparent lenses, the relative amount of trapped Chol increases with age in nuclear membranes. However, the EPR spectrum of ASL from cortical membranes of 15- to 60-year-old donors shows only the weakly immobilized component assigned to ASL in the bulk plus Chol bilayer domain. Only the cortical membranes of 61- to 70-year-old donors contain both weakly and strongly immobilized components. The strongly immobilized component is assigned to ASL in trapped lipids. We speculate that the age of 60 years may be considered as a "threshold" for appearance of trapped lipids in cortical membranes. The relative amounts of boundary plus trapped PLs in lenses with nuclear cataracts is lower than that predicted from the tendency of the age-dependent increase observed for transparent lenses. The differences in amounts of lipids in the indicated left and right eye domains of each donor are smaller than the differences in single donors of a similar age.

Interactions of beta-crystallin with a model of human eye lens-lipid membranes.

β-Crystallin helps maintain the eye lens's structure and refractivity. With age and cataract formation, α-, β-, and γ-crystallin may undergo various mutations and post-translational modifications, and consequently form higher molecular weight complexes, some of which bind to lens membranes. However, the binding of β-crystallin alone to the lens membrane is still unclear. This study aims to investigate the binding of β-crystallin to the model of the human lens-lipid (MHLL) membrane and the physical properties of the MHLL membrane after β-crystallin binding. Small unilamellar vesicles of MHLL membranes, resembling the human lens-lipid composition, were prepared using the rapid solvent exchange method and probe-tip sonication. The 1 mol% cholesterol analogue spin label (CSL) integrated into the membrane, which resides near the lens-lipid headgroup region, probes β-crystallin binding to the membrane using the electron paramagnetic resonance (EPR) spin-labeling method. Our results showed that β-crystallin binds to the MHLL membrane with a binding affinity (Ka) of 0.082 ± 0.006 μM−1 and saturates around 5% of the membrane surface occupied. Furthermore, β-crystallin binding to the MHLL membrane decreased the mobility parameter, making the headgroups of the membrane less mobile. However, β-crystallin binding to the MHLL membrane showed no significant change in maximum splitting, meaning it does not affect the order near the headgroup regions of the MHLL membrane. Moreover, the hydrophobicity of the MHLL membrane also did not change significantly with the binding of β-crystallin, meaning hydrophobicity near the headgroup regions does not significantly change with β-crystallin binding. Our results showed that β-crystallin binding with MHLL membranes changes the physical property of membranes, implying that β-crystallin binding to the lens membrane may change the integrity of the membrane and has the potential to promote cataract formation.

A sensitive method to investigate the interaction of alpha-crystallin with cortical and nuclear lens lipid membranes derived from a single lens.

The levels of α-crystallin in eye lens cytoplasm decrease with a proportional increase in membrane-bound α-crystallin with age and cataract formation. The influence of differences in lipids composition between cortical membrane (CM) and nuclear membrane (NM) derived from a single lens on α-crystallin binding to these membranes is still unclear. This study aims to develop a sensitive electron paramagnetic resonance (EPR) spin-labeling method to investigate the binding of α-crystallin to the CM and NM derived from total lipids extracted from a single lens of a bovine. No binding of α-crystallin below the surface of CM and small binding of α-crystallin below the surface of NM was observed using cholesterol analogue spin-label (CSL) within these membranes. However, significant binding of α-crystallin on the surface of CM and NM was observed using 4-palmitamido-TEMPO (4PT) spin-label within these membranes. The percentage of membrane surface occupied and the binding affinity for NM are higher than for CM, correlating with less mobility and more order below and on the surface of NM than CM. The mobility decreases with no significant change in order and hydrophobicity below and on the surface of CM and NM with α-crystallin binding. Our results imply that α-crystallin primarily binds on the surface of bovine CM and NM, and such surface binding of α-crystallin to membranes in young and clear lenses may be helpful for membrane stability. However, α-crystallin binding increases hydrophobicity below the surface of the cortical membrane with low cholesterol (CMLC), mimicking the decreased cholesterol content in cataractous lenses, suggesting that α-crystallin binding below the surface of the CMLC forms a hydrophobic barrier for the passage of polar molecules supporting the barrier hypothesis in cataract formation.

MR vessel-encoded arterial spin labeling with the placement of metallic items to visualize the territorial blood flow after extracranial-intracranial bypass surgery: a proof-of-concept study.

Depiction of bypass blood flow in patients who received extracranial-intracranial (EC-IC) bypass surgery is important for patient care. To develop a vessel-encoded arterial spin labeling (VE-ASL) method using surgical staples as a magnetic resonance (MR)-conditional product in patients who received EC-IC bypass surgery. Pseudo-continuous labeling was used for VE-ASL acquisition with a 3-T MR unit. First, an experimental study was conducted to determine the appropriate number of surgical staples to obtain a spatially sufficient saturation effect. Thereafter, four healthy normal volunteers underwent a VE-ASL study to confirm the sufficiency of the saturation effect to the right or left common carotid artery. Finally, VE-ASL scanning was performed in seven patients after EC-IC bypass surgery to confirm the ability of VE-ASL to visualize the territorial bypass perfusion. All qualitative evaluation was performed by two neuroradiologists using a 3-point grading system (2 = good, 1 = moderate, 0 = poor). A quantity of 200 staples was found to be appropriate for VE-ASL scanning. In healthy volunteers, one neuroradiologist rated the images of all four cases as good, while the other rated three cases as good and one case as moderate. For the seven patients after EC-IC bypass surgery, one neuroradiologist rated all seven cases as good, and the other rated six cases as good and one case as moderate. VE-ASL using surgical staples might be useful for the evaluation of territorial bypass perfusion in patients after EC-IC bypass surgery.

PS-C05-12: OMENTIN-1 PREDICTS MEMBRANE FLUIDITY OF RED BLOOD CELLS AND MICROCIRCULATORY FUNCTION IN HYPERTENSIVE SUBJECTS BY THE NITRIC OXIDE-DEPENDENT MECHANISM

Objective: It has been shown that abnormalities in physical properties of the cell membranes may strongly be linked to hypertension. Recent evidence indicates that omentin-1, an adipokine with anti-inflammatory properties, may actively participate in the pathophysiology of hypertension, atherosclerosis and other cardiovascular diseases. However, few studies have evaluated the association between omentin-1 and membrane function in hypertension. The present study was performed to investigate possible relationships among omentin-1, endothelial function and membrane fluidity (a reciprocal value of membrane microviscosity) of red blood cells (RBCs) in patients with essential hypertension and normotensive subjects. Design and method: We measured the membrane fluidity of RBCs with an electron spin resonance (ESR) and spin labeling method. In each ESR spectrum, we calculated the order parameter (S) for the spin-label agent (5-nitroxide stearate). The greater the value of the order parameter (S) was, the lower the membrane fluidity of RBC was. Results: (1) The order parameter (S) obtained from the ESR spectra of RBC membranes was significantly greater in hypertensive subjects (0.727 ± 0.001, n = 64) than in normotensive subjects (0.719 ± 0.001, n = 49, P &lt; 0.0001), indicating that membrane fluidity of RBCs might be reduced in hypertensive subjects compared with normotensive subjects. (2) Plasma omentin-1 levels were lower in hypertensive subjects than in normotensive subjects (HT 0.486 ± 0.014 ng/mL, n = 64, NT 0.537 ± 0.019 ng/mL, n = 49, P &lt; 0.05). Plamsa omentin-1 significantly correlated with plasma nitric oxide (NO)-metabolite levels (an index of endothelial function) (r = 0.392, n = 113, P &lt; 0.0001). (3) Furthermore, the order parameter (S) of RBCs inversely correlated with both plasma omentin-1 (r = -0.347, n = 113, P &lt; 0.001) and plasma NO-metabolites, suggesting that reduced membrane fluidity of RBCs might be associated with decreased levels of plasma omentin-1 and endothelial dysfunction. (4) Multivariate regression analysis showed that, after adjusting for confounding factors, plasma omentin-1 might be an independent predictor of membrane fluidity of RBCs. Conclusions: The ESR study demonstrated that lower plasma omentin-1 levels might have a close correlation with reduced membrane fluidity of RBCs and impaired hemorheologic behavior in hypertensive subjects by the NO-dependent mechanism. It is strongly suggested that plasma omentin-1 could be a putative biomarker for the microcirculatory function in hypertension.

Robust dual-module velocity-selective arterial spin labeling (dm-VSASL) with velocity-selective saturation and inversion.

Compared to conventional arterial spin labeling (ASL) methods, velocity-selective ASL (VSASL) is more sensitive to artifacts from eddy currents, diffusion attenuation, and motion. Background suppression is typically suboptimal in VSASL, especially of CSF. As a result, the temporal SNR and quantification accuracy of VSASL are compromised, hindering its application despite its advantage of being delay-insensitive. A novel dual-module VSASL (dm-VSASL) strategy is developed to improve the SNR efficiency and the temporal SNR with a more balanced gradient configuration in the label/control image acquisition. This strategy applies for both VS saturation (VSS) and VS inversion (VSI) labeling. The dm-VSASL schemes were compared with single-module labeling and a previously developed multi-module schemes for the SNR performance, background suppression efficacy, and sensitivity to artifacts in simulation and in vivo experiments, using pulsed ASL as the reference. Dm-VSASL enabled more robust labeling and efficient backgroud suppre across brain tissues, especially of CSF, resulting in significantly reduced artifacts and improved temporal SNR. Compared to single-module labeling, dm-VSASL significantly improved the temporal SNR in gray (by 90.8% and 94.9% for dm-VSS and dm-VSI, respectively; P< 0.001) and white (by 41.5% and 55.1% for dm-VSS and dm-VSI, respectively; P< 0.002) matter. Dm-VSI also improved the SNR of VSI by 5.4% (P= 0.018). Dm-VSASL can significantly improve the robustness of VS labeling, reduce artifacts, and allow efficient background suppression. When implemented with VSI, it provides the highest SNR efficiency among VSASL methods. Dm-VSASL is a powerful ASL method for robust, accurate, and delay-insensitive perfusion mapping.

Binding of Alpha-Crystallin to Cortical and Nuclear Lens Lipid Membranes Derived from a Single Lens.

Several studies reported that α-crystallin concentrations in the eye lens cytoplasm decrease with a corresponding increase in membrane-bound α-crystallin with age and cataracts. The influence of the lipid and cholesterol composition difference between cortical membrane (CM) and nuclear membrane (NM) on α-crystallin binding to membranes is still unclear. This study uses the electron paramagnetic resonance (EPR) spin-labeling method to investigate the α-crystallin binding to bovine CM and NM derived from the total lipids extracted from a single lens. Compared to CMs, NMs have a higher percentage of membrane surface occupied by α-crystallin and binding affinity, correlating with less mobility and more order below and on the surface of NMs. α-Crystallin binding to CM and NM decreases mobility with no significant change in order and hydrophobicity below and on the surface of membranes. Our results suggest that α-crystallin mainly binds on the surface of bovine CM and NM and such surface binding of α-crystallin to membranes in clear and young lenses may play a beneficial role in membrane stability. However, with decreased cholesterol content within the CM, which mimics the decreased cholesterol content in the cataractous lens membrane, α-crystallin binding increases the hydrophobicity below the membrane surface, indicating that α-crystallin binding forms a hydrophobic barrier for the passage of polar molecules, supporting the barrier hypothesis in developing cataracts.

A preliminary study on predictors of treatment response to repetitive transcranial magnetic stimulation in patients with treatment-resistant depression in Japan.

Brain imaging studies have reported that the effect of repetitive transcranial magnetic stimulation (rTMS) is associated with the activities of the dorsolateral prefrontal cortex (DLPFC) and ventral medial prefrontal cortex (VMPFC). However, few studies have been conducted in Japanese patients. We aimed to identify brain regions associated with depressive symptom changes by measuring regional cerebral blood flow (rCBF) in the DLPFC and VMPFC before and after the high-frequency rTMS to the left DLPFC in Japanese patients with treatment-resistant depression. Fourteen patients participated in the rTMS study and were assessed with the 17-item Hamilton depression rating scale (HAM-D17 ). Among them, 13 participants underwent magnetic resonance imaging scan of the brain using the arterial spin labeling method. The rCBF was calculated using the fine stereotactic region of interest template (FineSRT) program for automated analysis. We focused on eight regions reported in previous studies. Depression severity significantly decreased after 2 week (HAM-D17 :11.4 ± 2.8, P=0.00027) and 4 week (HAM-D17 : 11.0 ± 3.7, P=0.0023) of rTMS treatment. There was no significant change in rCBF at each region in the pre-post design. However, there was a significantly negative correlation between baseline rCBF in the right DLPFC and the improvement in HAM-D17 score (r=-0.559, P=0.047). We obtained supportive evidence for the effectiveness of rTMS to the prefrontal cortex in treatment-resistant depression, which may be associated with reduced rCBF of the right DLPFC before initiation of rTMS.

Recent Technical Developments in ASL: A Review of the State of the Art.

This review article provides an overview of a range of recent technical developments in advanced arterial spin labeling (ASL) methods that have been developed or adopted by the community since the publication of a previous ASL consensus paper by Alsop et al. It is part of a series of review/recommendation papers from the International Society for Magnetic Resonance in Medicine Perfusion Study Group. Here, we focus on advancements in readouts and trajectories, image reconstruction, noise reduction, partial volume correction, quantification of nonperfusion parameters, fMRI, fingerprinting, vessel selective ASL, angiography, deep learning, and ultrahigh field ASL. We aim to provide a high level of understanding of these new approaches and some guidance for their implementation, with the goal of facilitating the adoption of such advances by research groups and by MRI vendors. Topics outside the scope of this article that are reviewed at length in separate articles include velocity selective ASL, multiple‐timepoint ASL, body ASL, and clinical ASL recommendations.

Abnormal Cerebral Blood Flow and Volumetric Brain Morphometry in Patients With Obstructive Sleep Apnea.

Obstructive sleep apnea (OSA) is a serious breathing disorder, leading to myocardial infarction, high blood pressure, and stroke. Brain morphological changes have been widely reported in patients with OSA. The pathophysiological mechanisms of cerebral blood flow (CBF) changes associated with OSA are not clear. In this study, 20 patients with OSA and 36 healthy controls (HCs) were recruited, and then pseudo-continuous arterial spin labeling (pCASL) and voxel-based morphometry (VBM) methods were utilized to explore blood perfusion and morphological changes in the patients with OSA. Compared with the HC group, the OSA group showed increased CBF values in the right medial prefrontal cortex (mPFC), left precentral gyrus, and right insula and showed decreased CBF values in the right temporal pole (TP) and the right cerebellum_Crus2. Compared with the HC group, the patients with OSA showed decreased gray matter volume (GMV) in the right dorsal lateral prefrontal cortex (DLPFC), the right occipital pole, and the vermis. There were no significantly increased GMV brain regions found in patients with OSA. Pearson correlation analysis showed that the reduced GMV in the right DLPFC and the right occipital pole was both positively correlated with Mini-Mental State Examination (MMSE) (r = 0.755, p < 0.001; r = 0.686, p = 0.002) and Montreal Cognitive Assessment (MoCA) scores (r = 0.716, p = 0.001; r = 0.601, p = 0.008), and the reduced GMV in the right occipital pole was negatively correlated with duration of illness (r = −0.497, p = 0.036). Patients with OSA have abnormal blood perfusion metabolism and morphological changes in brain regions including the frontal lobe and the cerebellum and were closely related to abnormal behavior, psychology, and cognitive function, which play an important role in the pathophysiological mechanism of OSA.

Multilamellar Liposomes as a Model for Biological Membranes: Saturation Recovery EPR Spin-Labeling Studies.

EPR spin labeling has been used extensively to study lipids in model membranes to understand their structures and dynamics in biological membranes. The lipid multilamellar liposomes, which are the most commonly used biological membrane model, were prepared using film deposition methods and investigated with the continuous wave EPR technique (T2-sensitive spin-labeling methods). These investigations provided knowledge about the orientation of lipids, their rotational and lateral diffusion, and their rate of flip-flop between bilayer leaflets, as well as profiles of membrane hydrophobicity, and are reviewed in many papers and book chapters. In the early 1980s, the saturation recovery EPR technique was introduced to membrane studies. Numerous T1-sensitive spin-label methods were developed to obtain detailed information about the three-dimensional dynamic membrane structure. T1-sensitive methods are advantageous over T2-sensitive methods because the T1 of spin labels (1–10 μs) is 10 to 1000 times longer than the T2, which allows for studies of membrane dynamics in a longer time–space scale. These investigations used multilamellar liposomes also prepared using the rapid solvent exchange method. Here, we review works in which saturation recovery EPR spin-labeling methods were applied to investigate the properties of multilamellar lipid liposomes, and we discuss their relationships to the properties of lipids in biological membranes.

Arterial Spin Labeling Perfusion in Determining the IDH1 Status and Ki-67 Index in Brain Gliomas.

The aim of the study was to evaluate the relationship between tumor blood flow (TBF) measured by the pseudo-continuous arterial spin labeling (PCASL) method and IDH1 mutation status of gliomas as well as Ki-67 proliferative index. Methods. The study included 116 patients with newly diagnosed gliomas of various grades. They received no chemotherapy or radiotherapy before MRI. IDH1 status assessment was performed after tumor removal in 106 cases—48 patients were diagnosed with wildtype gliomas (Grade 1–2—6 patients, Grade 3–4—42 patients) and 58 patients were diagnosed with mutant forms of gliomas (Grade 1–2—28 patients, Grade 3–4—30 patients). In 64 cases out of 116 Ki-67 index was measured. Absolute and normalized tumor blood flow values were measured on 3D PCASL maps. Results. TBF and normalized TBF (nTBF) in wildtype gliomas were significantly higher than in IDH1-mutant gliomas (p < 0.001). ASL perfusion showed high values of sensitivity and specificity in the differential diagnosis of gliomas with distinct IDH1 status (for TBF: specificity 75%, sensitivity 77.6%, AUC 0.783, cutoff 80.57 mL/100 g/min, for nTBF: specificity 77.1%, sensitivity 79.3%, AUC 0.791, cutoff 4.7). TBF and nTBF in wildtype high-grade gliomas (HGG) were significantly higher than in mutant forms (p < 0.001). ASL perfusion showed the following values of sensitivity and specificity in the diagnosis of mutant HGG and wildtype HGG (for TBF: specificity 83.3%, sensitivity 60%, AUC 0.719, cutoff 84.18 mL/100 g/min, for nTBF: specificity 88.1%, sensitivity 60%, AUC 0.729, cutoff 4.7). There was a significant positive correlation between tumor blood flow and Ki-67 (for TBF Rs = 0.63, for nTBF Rs = 0.61). Conclusion. ASL perfusion may be an informative factor in determining the IDH1 status in brain gliomas preoperative and tumor proliferative activity.

Phospholipid acyl chain length and degree of unsaturation modulate α-crystallin binding to the membrane

The binding of α-crystallin to the eye lens’s fiber cell plasma membrane increases with age and cataract. It has also been reported that the primary binding site of α-crystallin in the lens’s membrane is phospholipids (PLs). However, it is unclear whether PL acyl chain length and degree of unsaturation influence α-crystallin binding. We have used electron paramagnetic resonance spin-labeling methods to investigate the binding of α-crystallin with cholesterol (Chol)/phosphatidylcholine (PC) membrane of different chain lengths and degree of unsaturation (i.e., 14:0 PC, 18:0 18:1 PC, 18:1 PC, and 16:0 20:4 PC) and in the presence and absence of 23 mol% Chol.