Neurocognition and brain functional connectivity in a non-clinical population-based sample with psychotic experiences.

We characterized the neurocognitive profile of communed-based individuals and unaffected siblings of patients with psychosis from Brazil reporting psychotic experiences (PEs). We also analyzed associations between PEs and the intra and inter-functional connectivity (FC) in the Default Mode Network (DMN), the Fronto-Parietal Network (FPN) and the Salience Network (SN) measured by functional magnetic resonance imaging. The combined sample of communed-based individuals and unaffected siblings of patients with psychosis comprised 417 (neurocognition) and 85 (FC) volunteers who were divided as having low (<75th percentile) and high (≥75th percentile) PEs (positive, negative, and depressive dimensions) assessed by the Community Assessment of Psychic Experiences. The neurocognitive profile and the estimated current brief intellectual quotient (IQ) were assessed using the digit symbol (processing speed), arithmetic (working memory), block design (visual learning) and information (verbal learning) subtests of Wechsler Adult Intelligence Scale-third edition. Logistic regression models were performed for neurocognitive analysis. For neuroimaging, we used the CONN toolbox to assess FC between the specified regions, and ROI-to-ROI analysis. In the combined sample, high PEs (all dimensions) were related to lower processing speed performance. High negative PEs were related to poor visual learning performance and lower IQ, while high depressive PEs were associated with poor working memory performance. Those with high negative PEs presented FPN hypoconnectivity between the right and left lateral prefrontal cortex. There were no associations between PEs and the DMN and SN FC. Brazilian individuals with high PEs showed neurocognitive impairments like those living in wealthier countries. Hypoconnectivity in the FPN in a community sample with high PEs is coherent with the hypothesis of functional dysconnectivity in schizophrenia.

Influence of cerebral blood flow on volumetric loss related to Alzheimer's disease.

CBF measured with Arterial Spin Labeling (ASL) obtained by Magnetic Resonance Imaging (MRI) may become an important biomarker by showing changes in early stages of AD, such as in the prodromal stage of Mild Cognitive Impairment (MCI). Objective: Verify the correlation between atrophy and CBF in patients with MCI and mild phase ADD, to demonstrate whether changes in CBF can be considered as vascular biomarkers in the diagnosis of the DA continuum. Methods: 11 healthy volunteers, 16 MCI and 15 mild ADD were evaluated. Images of the brain were acquired, including CBF measured with Arterial Spin Labeling (ASL). Results: When comparing MCI with control, a reduction in normalized CBF was observed in left posterior cingulate (estimated difference -0.38; p=0.02), right posterior cingulate (estimated difference -0.45; p=0.02) and right precuneus (estimated difference -0.28; p <0.01); also increase in normalized CBF in right upper temporal pole (estimated difference 0.22; p=0.03). It was also observed that in MCI, the smaller the gray matter volume, the smaller the CBF in the left posterior cingulate; as well as the greater the cerebrospinal fluid volume, consequent to the encephalic volumetric reduction associated with atrophy, the greater the CBF in the right superior temporal pole. When comparing controls, MCI and mild AD, in relation to the other variables, no other correlations were observed between CBF and atrophy. Conclusion: In patients with MCI, the reduction of CBF in the left posterior cingulate correlated with gray matter atrophy, as well as the increase of CBF in the right upper temporal pole correlated with an increase in cerebrospinal fluid consequent to the encephalic volumetric reduction associated with atrophy, demonstrating the influence of CBF in AD related brain atrophy. These findings position CBF as a possible vascular biomarker for early-stage AD diagnoses.

A imagem por ressonância magnética (IRM) pode se tornar um importante biomarcador ao mostrar alterações nos estágios iniciais da doença de Alzheimer (DA). Objetivo: Sendo a atrofia cerebral um importante biomarcador de neurodegeneração na DA, o presente estudo foi realizado com o objetivo de verificar se há correlação entre atrofia e fluxo sanguíneo cerebral (FSC) em pacientes com diagnóstico de CCL e demência da doença de Alzheimer (DDA) leve, com o objetivo de revelar se as alterações no FSC podem ser consideradas possíveis biomarcadores vasculares no diagnóstico do continuum da DA. Métodos: Foram avaliados 11 voluntários saudáveis, 16 CCL e 15 DDA leve. Imagens do cérebro foram adquiridas em um equipamento de 3 T, incluindo imagens ponderadas em T1 de alta resolução para avaliação anatômica e Arterial Spin Labeling (ASL) para a quantificação de FSC. Resultados: Quando comparado CCL com controle, observou-se redução no FSC normalizado em cingulado posterior esquerdo (diferença estimada de -0,38; p=0,02), cingulado posterior direito (diferença estimada de -0,45; p=0,02) e precúneo direito (diferença estimada de -0,28; p <0,01); e aumento de FSC normalizado no polo temporal superior direito (diferença estimada de 0,22; p=0,03). No CCL, quanto menor o volume da substância cinzenta, menor o FSC no cingulado posterior esquerdo; quanto maior o volume de fluido cerebroespinhal, consequente à redução volumétrica encefálica, maior o FSC no polo temporal superior direito. Conclusão: Nos pacientes com diagnóstico de CCL, a redução de FSC no cingulado posterior esquerdo apresentou correlação com atrofia da substância cinzenta, assim como o aumento de FSC no polo temporal superior direito apresentou correlação com o aumento de fluido cerebroespinhal, demonstrando a provável influência do FSC na atrofia encefálica relacionada à DA.

Cortical Thickness and Brain Connectivity Mediate the Relation Between White Matter Hyperintensity and Information Processing Speed in Cerebral Small Vessel Disease.

White matter hyperintensities of presumed vascular origin (WMH) are the most common imaging feature of cerebral small vessel disease (cSVD) and are associated with cognitive impairment, especially information processing speed (IPS) deficits. However, it is unclear how WMH can directly impact IPS or whether the cortical thickness and brain connectivity mediate such association. In this study, it was evaluated the possible mediating roles of cortical thickness and brain (structural and functional) connectivity on the relationship between WMH (also considering its topography distribution) and IPS in 389 patients with cSVD from the RUN-DMC (Radboud University Nijmegen Diffusion tensor and Magnetic resonance imaging Cohort) database. Significant (p < 0.05 after multiple comparisons correction) associations of WMH volume and topography with cortical thickness, brain connectivity, and IPS performance in cSVD individuals were found. Additionally, cortical thickness and brain structural and functional connectivity were shown to mediate the association of WMH volume and location with IPS scores. More specifically, frontal cortical thickness, functional sensorimotor network, and posterior thalamic radiation tract were the essential mediators of WMH and IPS in this clinical group. This study provided insight into the mechanisms underlying the clinical relevance of white matter hyperintensities in information processing speed deficits in cSVD through cortical thinning and network disruptions.

Neural Substrates of Psychomotor Speed Deficits in Cerebral Small Vessel Disease: A Brain Disconnectome Mapping Study.

It remains unknown which factors influence how brain disconnectivity derived from White Matter Hyperintensity (WMH) lesions leads to psychomotor speed dysfunction, one of the earliest and most common cognitive manifestations in the cerebral Small Vessel Disease (cSVD) population. While the burden of WMH has been strongly linked to psychomotor speed performance, the effect that different locations and volumes of WMH may have on cSVD-related cognitive impairment remains unclear. Therefore, we aimed to explore (1) whether global WMH, deep WMH (DWMH), and periventricular (PVWMH) volumes display different psychomotor speed associations; (2) whether tract-specific WMH volume shows stronger cognitive associations compared with global measures of WMH volume; (3) whether specific patterns of WMH location lead to different degrees of disconnectivity. Using the BCBToolkit, we investigated which pattern of distribution and which locations of WMH lesion result in impaired psychomotor speed in a well-characterized sample (n = 195) of cSVD patients without dementia. Two key findings emerge from our study. First, global (and not tract-specific) measures of WMH volume were associated with psychomotor speed performance. Second, disconnection maps revealed the involvement of callosal tracts, association and projection fibers, and frontal and parietal cortical brain areas related to psychomotor speed, while the lesion location influenced such associations. In conclusion, psychomotor deficits are affected differently by WMH burden and topographic distribution through brain disconnection in non-demented cSVD patients.

Influence of cerebral blood flow on volumetric loss related to Alzheimer's disease / Influência do fluxo sanguíneo cerebral na perda volumétrica relacionada à doença de Alzheimer

ABSTRACT: CBF measured with Arterial Spin Labeling (ASL) obtained by Magnetic Resonance Imaging (MRI) may become an important biomarker by showing changes in early stages of AD, such as in the prodromal stage of Mild Cognitive Impairment (MCI). Objective: Verify the correlation between atrophy and CBF in patients with MCI and mild phase ADD, to demonstrate whether changes in CBF can be considered as vascular biomarkers in the diagnosis of the DA continuum. Methods: 11 healthy volunteers, 16 MCI and 15 mild ADD were evaluated. Images of the brain were acquired, including CBF measured with Arterial Spin Labeling (ASL). Results: When comparing MCI with control, a reduction in normalized CBF was observed in left posterior cingulate (estimated difference -0.38; p=0.02), right posterior cingulate (estimated difference -0.45; p=0.02) and right precuneus (estimated difference -0.28; p <0.01); also increase in normalized CBF in right upper temporal pole (estimated difference 0.22; p=0.03). It was also observed that in MCI, the smaller the gray matter volume, the smaller the CBF in the left posterior cingulate; as well as the greater the cerebrospinal fluid volume, consequent to the encephalic volumetric reduction associated with atrophy, the greater the CBF in the right superior temporal pole. When comparing controls, MCI and mild AD, in relation to the other variables, no other correlations were observed between CBF and atrophy. Conclusion: In patients with MCI, the reduction of CBF in the left posterior cingulate correlated with gray matter atrophy, as well as the increase of CBF in the right upper temporal pole correlated with an increase in cerebrospinal fluid consequent to the encephalic volumetric reduction associated with atrophy, demonstrating the influence of CBF in AD related brain atrophy. These findings position CBF as a possible vascular biomarker for early-stage AD diagnoses.

RESUMO: A imagem por ressonância magnética (IRM) pode se tornar um importante biomarcador ao mostrar alterações nos estágios iniciais da doença de Alzheimer (DA). Objetivo: Sendo a atrofia cerebral um importante biomarcador de neurodegeneração na DA, o presente estudo foi realizado com o objetivo de verificar se há correlação entre atrofia e fluxo sanguíneo cerebral (FSC) em pacientes com diagnóstico de CCL e demência da doença de Alzheimer (DDA) leve, com o objetivo de revelar se as alterações no FSC podem ser consideradas possíveis biomarcadores vasculares no diagnóstico do continuum da DA. Métodos: Foram avaliados 11 voluntários saudáveis, 16 CCL e 15 DDA leve. Imagens do cérebro foram adquiridas em um equipamento de 3 T, incluindo imagens ponderadas em T1 de alta resolução para avaliação anatômica e Arterial Spin Labeling (ASL) para a quantificação de FSC. Resultados: Quando comparado CCL com controle, observou-se redução no FSC normalizado em cingulado posterior esquerdo (diferença estimada de -0,38; p=0,02), cingulado posterior direito (diferença estimada de -0,45; p=0,02) e precúneo direito (diferença estimada de -0,28; p <0,01); e aumento de FSC normalizado no polo temporal superior direito (diferença estimada de 0,22; p=0,03). No CCL, quanto menor o volume da substância cinzenta, menor o FSC no cingulado posterior esquerdo; quanto maior o volume de fluido cerebroespinhal, consequente à redução volumétrica encefálica, maior o FSC no polo temporal superior direito. Conclusão: Nos pacientes com diagnóstico de CCL, a redução de FSC no cingulado posterior esquerdo apresentou correlação com atrofia da substância cinzenta, assim como o aumento de FSC no polo temporal superior direito apresentou correlação com o aumento de fluido cerebroespinhal, demonstrando a provável influência do FSC na atrofia encefálica relacionada à DA.

Contrast agent-free state-of-the-art magnetic resonance imaging on cerebral small vessel disease - Part 2: Diffusion tensor imaging and functional magnetic resonance imaging.

Cerebral small vessel disease (cSVD) has been widely studied using conventional magnetic resonance imaging (MRI) methods, although the association between MRI findings and clinical features of cSVD is not always concordant. We assessed the additional contribution of contrast agent-free, state-of-the-art MRI techniques, particularly diffusion tensor imaging (DTI) and functional magnetic resonance imaging (fMRI), to understand brain damage and structural and functional connectivity impairment related to cSVD. We performed a review following the PICOS worksheet and Search Strategy, including 152 original papers in English, published from 2000 to 2022. For each MRI method, we extracted information about their contributions regarding the origins, pathology, markers, and clinical outcomes in cSVD. In general, DTI studies have shown that changes in mean, radial, and axial diffusivity measures are related to the presence of cSVD. In addition to the classical deficit in executive functions and processing speed, fMRI studies indicate connectivity dysfunctions in other domains, such as sensorimotor, memory, and attention. Neuroimaging metrics have been correlated with the diagnosis, prognosis, and rehabilitation of patients with cSVD. In short, the application of contrast agent-free, state-of-the-art MRI techniques has provided a complete picture of cSVD markers and tools to explore questions that have not yet been clarified about this clinical condition. Longitudinal studies are desirable to look for causal relationships between image biomarkers and clinical outcomes.

Contrast-agent-free state-of-the-art MRI on cerebral small vessel disease-part 1. ASL, IVIM, and CVR.

Cerebral small vessel disease (cSVD), a common cause of stroke and dementia, is traditionally considered the small vessel equivalent of large artery occlusion or rupture that leads to cortical and subcortical brain damage. Microvessel endothelial dysfunction can also contribute to it. Brain imaging, including MRI, is useful to show the presence of lesions of several types, although the association between conventional MRI measures and clinical features of cSVD is not always concordant. We assessed the additional contribution of contrast-agent-free, state-of-the-art MRI techniques such as arterial spin labeling (ASL), diffusion tensor imaging, functional MRI, and intravoxel incoherent motion (IVIM) applied to cSVD in the existing literature. We performed a review following the PICO Worksheet and Search Strategy, including original papers in English, published between 2000 and 2022. For each MRI method, we extracted information about their contributions, in addition to those established with traditional MRI methods and related information about the origins, pathology, markers, and clinical outcomes in cSVD. This paper presents the first part of the review, which includes 37 studies focusing on ASL, IVIM, and cerebrovascular reactivity (CVR) measures. In general, they have shown that, in addition to white matter hyperintensities, alterations in other neuroimaging parameters such as blood flow and CVR also indicate the presence of cSVD. Such quantitative parameters were also related to cSVD risk factors. Therefore, they are promising, noninvasive tools to explore questions that have not yet been clarified about this clinical condition. However, protocol standardization is essential to increase their clinical use.

Feasibility of intravoxel incoherent motion in the assessment of tumor microvasculature and blood-brain barrier integrity: a case-based evaluation of gliomas.

OBJECTIVE: To evaluate the feasibility of intravoxel incoherent motion (IVIM) in assessing blood-brain barrier (BBB) integrity and microvasculature in tumoral tissue of glioma patients. METHODS: Images from 8 high-grade and 4 low-grade glioma patients were acquired on a 3 T MRI scanner. Acquisition protocol included pre- and post-contrast T1- and T2-weighted imaging, FLAIR, dynamic susceptibility contrast (DSC), and susceptibility-weighted imaging (SWI). In addition, IVIM was acquired with 15 b-values and fitted under the non-negative least square (NNLS) model to output the diffusion (D) and pseudo-diffusion (D*) coefficients, perfusion fraction (f), and f times D* (fD*) maps. RESULTS: IVIM perfusion-related maps were sensitive to (1) blood flow and perfusion alterations within the microvasculature of brain tumors, in agreement with intra-tumoral susceptibility signal (ITSS); (2) enhancing areas of BBB breakdown in agreement with DSC maps as well as areas of BBB abnormality that was not detected on DSC maps; (3) enhancing perfusion changes within edemas; (4) detecting early foci of increased perfusion within low-grade gliomas. CONCLUSION: The results suggest IVIM may be a promising approach to delineate tumor extension and progression in size, and to predict histological grade, which are clinically relevant information that characterize tumors and guide therapeutic decisions in patients with glioma.

Advances in resting state fMRI acquisitions for functional connectomics.

Resting state functional magnetic resonance imaging (rs-fMRI) is based on spontaneous fluctuations in the blood oxygen level dependent (BOLD) signal, which occur simultaneously in different brain regions, without the subject performing an explicit task. The low-frequency oscillations of the rs-fMRI signal demonstrate an intrinsic spatiotemporal organization in the brain (brain networks) that may relate to the underlying neural activity. In this review article, we briefly describe the current acquisition techniques for rs-fMRI data, from the most common approaches for resting state acquisition strategies, to more recent investigations with dedicated hardware and ultra-high fields. Specific sequences that allow very fast acquisitions, or multiple echoes, are discussed next. We then consider how acquisition methods weighted towards specific parts of the BOLD signal, like the Cerebral Blood Flow (CBF) or Volume (CBV), can provide more spatially specific network information. These approaches are being developed alongside the commonly used BOLD-weighted acquisitions. Finally, specific applications of rs-fMRI to challenging regions such as the laminae in the neocortex, and the networks within the large areas of subcortical white matter regions are discussed. We finish the review with recommendations for acquisition strategies for a range of typical applications of resting state fMRI.

Semantic verbal fluency brain network: delineating a physiological basis for the functional hubs using dual-echo ASL and graph theory approach.

Objective. Semantic verbal fluency (SFV) is a cognitive process that engages and modulates specific brain areas related to language comprehension and production, decision making, response inhibition, and memory retrieval. The impairment of the brain network responsible for these functions is related to various neurological conditions, and different strategies have been proposed to assess SVF-related deficits in such diseases. In the present study, the concomitant changes of brain perfusion and functional connectivity were investigated during the resting state and SVF task performance.Approach. Arterial spin labeling (ASL), a perfusion-based magnetic resonance imaging (MRI) method, was used with a pseudocontinuous labeling approach and dual-echo readout in 28 healthy right-handed Brazilian Portuguese speakers. The acquisition was performed in a resting state condition and during the performance of a SVF task.Main results. During task performance, a significant increase in cerebral blood flow (CBF) was observed in language-related regions of the frontal lobe, including Brodmann's areas 6, 9, 45, and 47, associated with semantic processing, word retrieval, and speech motor programming. Such regions, along with the posterior cingulate, showed a crucial role in the SVF functional network, assessed by seed-to-voxel and graph analysis. Our approach successfully overcame the generalization problem regarding functional MRI (fMRI) graph analysis with cognitive, task-based paradigms. Moreover, the CBF maps enabled the functional assessment of orbital frontal and temporal regions commonly affected by magnetic susceptibility artifacts in conventional T2*-weighted fMRI approaches.Significance. Our results demonstrated the capability of ASL to evaluate perfusion alterations and functional patterns simultaneously regarding the SVF network providing a quantitative physiological basis to functional hubs in this network, which may support future clinical studies.

Functional and structural brain connectivity in congenital deafness.

Several studies have been carried out to verify neural plasticity and the language process in deaf individuals. However, further investigations regarding the intrinsic brain organization on functional and structural neural networks derived from congenital deafness are still an open question. The objective of this study was to investigate the main differences in brain organization manifested in congenitally deaf individuals, concerning the resting-state functional patterns, and white matter structuring. Functional and diffusion magnetic resonance imaging modalities were acquired from 18 congenitally deaf individuals and 18 age-sex-matched hearing controls. Compared to the hearing group, the deaf individuals presented higher functional connectivity among the posterior cingulate cortex node of the default mode network with visual and motor networks, lower functional connectivity between salience networks, language networks, and prominence of functional connectivity changes in the right hemisphere, mostly in the frontoparietal and temporal lobes. In terms of structural connectivity, we found changes mainly in the occipital and parietal lobes, involving both classical sign language support regions as well as concentrated networks for focus activity, attention, and cognitive filtering. Our findings demonstrated that the congenital deaf individuals who learned sign language developed significant brain functional and structural reorganization, which provides prominent support for large-scale brain networks associated with attention decision-making, environmental monitoring based on the movement of objects, and on the motor and visual controls.

[Formula: see text] Neuropsychological functioning and its relationship with brain anatomical measures of children and adolescents with non-syndromic cleft lip and palate.

BACKGROUND: Children and adolescents with non-syndromic cleft lip and palate (NSCLP) show cognitive performance below expected. This difficulty can be associated with alterations in the cortical thickness and volume of brain regions. The aim of this study was to investigate anatomical brain characteristics and their relationship with the neuropsychological scores of children and adolescents with NSCLP. Methods: Twenty-four children and adolescents with ages from 10 to 16 years and 11 months (12 with a diagnosis of NSCLP; 12 with typical development) were enrolled. Neuropsychological tests were administered and high-resolution, structural magnetic resonance imaging (MRI) was performed in a 1.5 T scanner. Results: Compared to the control group, NSCLP individuals showed intellectual (p = 0.006) and cognitive (p = 0.003) impairment, as well as deficits in subdomains of executive functions (sustained attention, working memory, and cognitive planning). The morphological analysis showed reduced volumes and cortical thickness in temporal, parietal, and frontal regions, in both hemispheres, of the NSCLP group. Significant, strong associations of structural alterations and cognitive performance were observed. Conclusions: Our study provided strong evidence of the relationship between brain development in children and adolescents with NSCLP, and their neuropsychological profile. This relationship is characterized by a malfunction of associative areas of the brain, such as parieto-temporo-occipital, frontoparietal, and prefrontal regions.

Contrast optimization in arterial spin labeling with multiple post-labeling delays for cerebrovascular assessment.

OBJECTIVE: Improving the readout for arterial spin labeling with multiple post-labeling delays (multi-PLD ASL) through a flip angle (FA) sweep towards increasing contrast-to-noise ratio for long PLD images. METHODS: Images were acquired from 20 healthy subjects and 14 patients with severe, asymptomatic carotid artery stenosis (ACAS) in a 3T MRI scanner. Multi-PLD ASL images with conventional and proposed (FA sweep) readouts were acquired. For patients, magnetic resonance angiography was used to validate the multi-PLD ASL results. Perfusion values were calculated for brain regions irrigated by the main cerebral arteries and compared by analysis of variance. RESULTS: For healthy subjects, better contrast was obtained for long PLDs when using the proposed multi-PLD method compared to the conventional. For both methods, no hemispheric difference of perfusion was observed. For patients, the proposed method facilitated the observation of delayed tissue perfusion, which was not visible for long PLD using the conventional multi-PLD ASL. CONCLUSION: We successfully assessed brain perfusion of patients with asymptomatic CAS using multi-PLD ASL with FA sweep. We were able to show subtle individual differences. Moreover, prolonged arterial transit time in patients was observed, although they were considered asymptomatic, suggesting that it may not be an adequate term to characterize them.

Brain Structural-Functional Connectivity Relationship Underlying the Information Processing Speed.

Human cognition and behavior emerge from neuronal interactions on a brain structural architecture. The convergence (or divergence) between functional dynamics and structural connectivity (SC) and their relationship with cognition are still a pivotal question about the brain. We focused on the information processing speed (IPS), assessed by the Symbol Digit Modalities Test (SDMT), once delayed IPS underlies attention deficits in various clinical conditions. We hypothesize that the SC constrains but does not determine functional connectivity, and such a relationship is related to the cognitive performance. Blood oxygenation level-dependent and diffusion tensor images of healthy young volunteers were acquired in a 3T magnetic resonance imaging machine. Activation maps included the left and right middle frontal gyri, left superior parietal lobule, left precuneus, left inferior frontal gyrus (IFG), right cuneus, left lingual gyrus, and left declive. A network involving such regions and signal propagation from visual, through cognitive, up to motor regions was proposed. Random effects Bayesian model selection showed that the top-down connections have the highest expected and exceedance probabilities. Moreover, all pairs of task-related regions were connected by at least one tract, except for the left declive with the left IFG. The interactions between the right cuneus with left declive were related to the interindividual variability in SDMT performance. Altogether, our findings suggest that the IPS functional network is related to the highest SDMT scores when its effective endogenous connections are suppressed to the detriment of modulation caused by the experimental conditions, with the underlying structure providing low diffusion environments.

Effects of global signal regression and subtraction methods on resting-state functional connectivity using arterial spin labeling data.

BACKGROUND: Arterial spin labeling (ASL) is an established magnetic resonance imaging (MRI) technique that is finding broader applications in functional studies of the healthy and diseased brain. To promote improvement in cerebral blood flow (CBF) signal specificity, many algorithms and imaging procedures, such as subtraction methods, were proposed to eliminate or, at least, minimize noise sources. Therefore, this study addressed the main considerations of how CBF functional connectivity (FC) is changed, regarding resting brain network (RBN) identification and correlations between regions of interest (ROI), by different subtraction methods and removal of residual motion artifacts and global signal fluctuations (RMAGSF). METHODS: Twenty young healthy participants (13 M/7F, mean age = 25 ±â€¯3 years) underwent an MRI protocol with a pseudo-continuous ASL (pCASL) sequence. Perfusion-based images were obtained using simple, sinc and running subtraction. RMAGSF removal was applied to all CBF time series. Independent Component Analysis (ICA) was used for RBN identification, while Pearson' correlation was performed for ROI-based FC analysis. RESULTS: Temporal signal-to-noise ratio (tSNR) was higher in CBF maps obtained by sinc subtraction, although RMAGSF removal had a significant effect on maps obtained with simple and running subtractions. Neither the subtraction method nor the RMAGSF removal directly affected the identification of RBNs. However, the number of correlated and anti-correlated voxels varied for different subtraction and filtering methods. In an ROI-to-ROI level, changes were prominent in FC values and their statistical significance. CONCLUSIONS: Our study showed that both RMAGSF filtering and subtraction method might influence resting-state FC results, especially in an ROI level, consequently affecting FC analysis and its interpretation. Taking our results and the whole discussion together, we understand that for an exploratory assessment of the brain, one could avoid removing RMAGSF to not bias FC measures, but could use sinc subtraction to minimize low-frequency contamination. However, CBF signal specificity and frequency range for filtering purposes still need to be assessed in future studies.