Designing and comparing optimized pseudo-continuous Arterial Spin Labeling protocols for measurement of cerebral blood flow

Arterial Spin Labeling (ASL) is a non-invasive, non-contrast, perfusion imaging technique which is inherently SNR limited. It is, therefore, important to carefully design scan protocols to ensure accurate measurements. Many pseudo-continuous ASL (PCASL) protocol designs have been proposed for measuring cerebral blood flow (CBF), but it has not yet been demonstrated which design offers the most accurate and repeatable CBF measurements. In this study, a wide range of literature PCASL protocols were first optimized for CBF accuracy and then compared using Monte Carlo simulations and in vivo experiments. The protocols included single-delay, sequential and time-encoded multi-timepoint protocols, and several novel protocol designs, which are hybrids of time-encoded and sequential multi-timepoint protocols. It was found that several multi-timepoint protocols produced more confident, accurate, and repeatable CBF estimates than the single-delay protocol, while also generating maps of arterial transit time. Of the literature protocols, the time-encoded protocol with T1-adjusted label durations gave the most confident and accurate CBF estimates in vivo (16% and 40% better than single-delay), while the sequential multi-timepoint protocol was the most repeatable (20% more repeatable than single-delay). One of the novel hybrid protocols, HybridT1-adj, was found to produce the most confident, accurate and repeatable CBF estimates out of all the protocols tested in both simulations and in vivo (24%, 47%, and 28% more confident, accurate, and repeatable than single-delay in vivo). The HybridT1-adj protocol makes use of the best aspects of both time-encoded and sequential multi-timepoint protocols and should be a useful tool for accurately and efficiently measuring CBF

A coupled drug kinetics-cell cycle model to analyse the response of human cells to intervention by topotecan

A model describing the response of the growth of single human cells in the absence and presence of the anti-cancer agent topotecan (TPT) is presented. The model includes a novel coupling of both the kinetics of TPT and cell cycle responses to the agent. By linking the models in this way, rather than using separate (disjoint) approaches, it is possible to illustrate how the drug perturbs the cell cycle. The model is compared to experimental in vitro cell cycle response data (comprising single cell descriptors for molecular and behavioural events), showing good qualitative agreement for a range of TPT dose levels

Model-based Bayesian inference of brain oxygenation using quantitative BOLD

© 2019 The Authors Streamlined Quantitative BOLD (sqBOLD) is an MR technique that can non-invasively measure physiological parameters including Oxygen Extraction Fraction (OEF) and deoxygenated blood volume (DBV) in the brain. Current sqBOLD methodology rely on fitting a linear model to log-transformed data acquired using an Asymmetric Spin Echo (ASE) pulse sequence. In this paper, a non-linear model implemented in a Bayesian framework was used to fit physiological parameters to ASE data. This model makes use of the full range of available ASE data, and incorporates the signal contribution from venous blood, which was ignored in previous analyses. Simulated data are used to demonstrate the intrinsic difficulty in estimating OEF and DBV simultaneously, and the benefits of the proposed non-linear model are shown. In vivo data are used to show that this model improves parameter estimation when compared with literature values. The model and analysis framework can be extended in a number of ways, and can incorporate prior information from external sources, so it has the potential to further improve OEF estimation using sqBOLD

Systems pharmacology approach for prediction of pulmonary and systemic pharmacokinetics and receptor occupancy of inhaled drugs

Pulmonary drug disposition after inhalation is complex involving mechanisms, such as regional drug deposition, dissolution, and mucociliary clearance. This study aimed to develop a systems pharmacology approach to mechanistically describe lung disposition in rats and thereby provide an integrated understanding of the system. When drug- and formulation-specific properties for the poorly soluble drug fluticasone propionate were fed into the model, it proved predictive of the pharmacokinetics and receptor occupancy after intravenous administration and nose-only inhalation. As the model clearly distinguishes among drug-specific, formulation-specific, and system-specific properties, it was possible to identify key determinants of pulmonary selectivity of receptor occupancy of inhaled drugs: slow particle dissolution and slow drug-receptor dissociation. Hence, it enables assessment of factors for lung targeting, including molecular properties, formulation, as well as the physiology of the animal species, thereby providing a general framework for rational drug design and facilitated translation of lung targeting from animal to man

A general framework for optimizing arterial spin labeling MRI experiments

PurposeArterial spin labeling (ASL) MRI is a non‐invasive perfusion imaging technique that is inherently SNR limited, so scan protocols ideally need to be rigorously optimized to provide the most accurate measurements. A general framework is presented for optimizing ASL experiments to achieve optimal accuracy for perfusion estimates and, if required, other hemodynamic parameters, within a fixed scan time. The effectiveness of this framework is then demonstrated by optimizing the post‐labeling delays (PLDs) of a multi‐PLD pseudo‐continuous ASL experiment and validating the improvement using simulations and in vivo data.Theory and MethodsA simple framework is proposed based on the use of the Cramér‐Rao lower bound to find the protocol design which minimizes the predicted parameter estimation errors. Protocols were optimized for cerebral blood flow (CBF) accuracy or both CBF and arterial transit time (ATT) accuracy and compared to a conventional multi‐PLD protocol, with evenly spaced PLDs, and a single‐PLD protocol, using simulations and in vivo experiments in healthy volunteers.ResultsSimulations and in vivo data agreed extremely well with the predicted performance of all protocols. For the in vivo experiments, optimizing for just CBF resulted in a 48% and 15% decrease in CBF errors, relative to the reference multi‐PLD and single‐PLD protocols, respectively. Optimizing for both CBF and ATT reduced CBF errors by 37%, without a reduction in ATT accuracy, relative to the reference multi‐PLD protocol.ConclusionThe presented framework can effectively design ASL experiments to minimize measurement errors based on the requirements of the scan

Pericyte heterogeneity identified by 3D ultrastructural analysis of the microvessel wall

Confident identification of pericytes (PCs) remains an obstacle in the field, as a single molecular marker for these unique perivascular cells remains elusive. Adding to this challenge is the recent appreciation that PC populations may be heterogeneous, displaying a range of morphologies within capillary networks. We found additional support on the ultrastructural level for the classification of these PC subtypes—“thin-strand” (TSP), mesh (MP), and ensheathing (EP)—based on distinct morphological characteristics. Interestingly, we also found several examples of another cell type, likely a vascular smooth muscle cell, in a medial layer between endothelial cells (ECs) and pericytes (PCs) harboring characteristics of the ensheathing type. A conserved feature across the different PC subtypes was the presence of extracellular matrix (ECM) surrounding the vascular unit and distributed in between neighboring cells. The thickness of this vascular basement membrane was remarkably consistent depending on its location, but never strayed beyond a range of 150–300 nm unless thinned to facilitate closer proximity of neighboring cells (suggesting direct contact). The density of PC-EC contact points (“peg-and-socket” structures) was another distinguishing feature across the different PC subtypes, as were the apparent contact locations between vascular cells and brain parenchymal cells. In addition to this thinning, the extracellular matrix (ECM) surrounding EPs displayed another unique configuration in the form of extensions that emitted out radially into the surrounding parenchyma. Knowledge of the origin and function of these structures is still emerging, but their appearance suggests the potential for being mechanical elements and/or perhaps signaling nodes via embedded molecular cues. Overall, this unique ultrastructural perspective provides new insights into PC heterogeneity and the presence of medial cells within the microvessel wall, the consideration of extracellular matrix (ECM) coverage as another PC identification criteria, and unique extracellular matrix (ECM) configurations (i.e., radial extensions) that may reveal additional aspects of PC heterogeneity

The RIG-I-like Receptor LGP2 Controls CD8+ T Cell Survival and Fitness

SummaryThe RIG-I-like receptors (RLRs) signal innate immune defenses upon RNA virus infection, but their roles in adaptive immunity have not been clearly defined. Here, we showed that the RLR LGP2 was not essential for induction of innate immune defenses, but rather was required for controlling antigen-specific CD8+ T cell survival and fitness during peripheral T cell-number expansion in response to virus infection. Adoptive transfer and biochemical studies demonstrated that T cell-receptor signaling induced LGP2 expression wherein LGP2 operated to regulate death-receptor signaling and imparted sensitivity to CD95-mediated cell death. Thus, LGP2 promotes an essential prosurvival signal in response to antigen stimulation to confer CD8+ T cell-number expansion and effector functions against divergent RNA viruses, including West Nile virus and lymphocytic choriomeningitis virus