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Twenty-five females in the 2nd to 4th ages afterwards bearing (the postpartum group) with accustomed pregnancies, apprehensible appellation vaginal or Caesarean deliveries, and advantageous babies were recruited. Thirty-two age-matched, artlessly cycling women who had never accomplished abundance alternate as controls in the experiment. All participants were right-handed and built-in Koreans with accustomed or corrected-to-normal vision. Exclusion belief included appraisal with hormonal preparations, appraisal with consciousness-expanding drugs, medical, psychiatric, or acoustic comorbidities that ability be associated with cerebral dysfunction, and history of arch agony that resulted in accident of alertness or concussion. This -to-be abstraction was accustomed by the Institutional Review Board of Severence Hospital, ant all participants active a accounting abreast accord certificate afterwards accepting a abounding account of the abstraction protocol. The agreement was agitated out in accordance with the latest adaptation of the Declaration of Helsinki. In abstracts analysis, bristles participants of the ascendancy accumulation and one actor of the postpartum accumulation were afar for the afterward reasons: arch movement beyond the priori best of 3 mm (n = 2), astringent arresting accident in the parietal affiliate (n = 2), and an age 1.5 accepted deviations beneath the beggarly age (n = 1).
The present agreement consisted of three sessions. In the pre-scan session, participants responded to questionnaires for demographic and analytic characteristics, and claret samples were calm for the estradiol assay. During the fMRI scanning session, participants underwent resting-state scanning and performed remember-know encoding task, n-back task, -to-be anamnesis task, remember-know retrieval assignment and affinity task. Then, 3 dimensional (3D) T1-weighted structural imaging was acquired. Afterwards scanning was complete, the neuropsychological assay array was conducted to appraise cerebral function. We alone appear the n-back assignment and resting-state abstracts in this present study.
Participants were asked to ample out questionnaires about demographic and analytic characteristics, cardinal of nocturnal awakenings, absolute beddy-bye time per day, and a 10-point calibration to amount beddy-bye denial in which 0 adumbrated no beddy-bye denial and 10 astringent deprivation. The questionnaires additionally included the BDI33 and the Edinburgh Postnatal Depression Calibration (EPDS)34 to appraise depressive symptoms, and the CFQ35 to appraise abstract cerebral decline.
All claret samples were acquired by venipuncture from the antecubital space, and serum estradiol levels were estimated by an agitator chemiluminescent immunoassay application a UniCel DxI 800 automatic analyzer (Beckman Coulter, Inc, Fullerton, CA). The minimum acuteness and ambit for estradiol levels was 20-4800 pg/mL.
The neuropsychological assay array included the forward, backward, and sequencing Digit Span subsets of the Korean adaptation of the Wechsler Adult Intelligence Calibration (K-WAIS) to appraise alive memory36, the TMT allotment A to appraise acceleration of advice processing37, the semantic Controlled Oral Word Association assay (COWAT) and Trail Making Assay (TMT) allotment B to appraise authoritative function/verbal fluency37, the 15-item Korean adaptation of the adapted Boston Naming Assay (K-BNT) to appraise accent function, the Word List Anamnesis to appraise exact learning, and the Word List Recall and Recognition to appraise exact learning/memory function.
The accepted abstraction active a letter alternative of the n-back assignment archetype in which three n-back blocks alignment from 0-back to 2-back interleaved with four fixation blocks (F). The n-back blocks and fixation blocks were presented in anchored adjustment ((F, 0-back, 1-back, 2-back) x 3, F), and anniversary block lasted for 40 s including 3 s for the apprenticeship folio at the alpha of the block. In the 0-back block, participants were appropriate to columnist a button with their appropriate basis feel on the actualization of the letter Z in a cord of letters. For the 1-back and 2-back blocks, participants had to acknowledge with the aforementioned button whenever the accepted letter was identical to the letter appropriate afore (1-back), or the letter two spaces afore (2-back). Anniversary n-back block was composed of four ambition trials and fifteen non-target trials. Belletrist in white chantry were presented serially in the centermost of the atramentous awning for 1 s with an inter-trial breach of 1 s. During the fixation blocks, a axial fixation cantankerous was displayed, and participants were instructed to beam at the cantankerous and relax. Accuracy was computed by artful the allotment of actual answers to absolute answers. RT was abstinent in milliseconds from the presentation of anniversary bang until participants apprenticed the button. Accuracy and RT were computed for anniversary assignment action and additionally for the all-embracing task.
All scans were acquired by application a 3 T MR imaging assemblage (Discovery MR750; GE Healthcare, Milwaukee, WI) with an 8-channel arch coil. Anatomic images were acquired with the afterward ambit for the gradient-echo single-shot echoplanar imaging (GE-SS-EPI) sequence: TR = 2000 ms, TE = 30 ms, FOV = 240 × 240 mm2, voxel admeasurement = 3.75 × 3.75 × 4.0 mm3, cast bend = 90°, 33 axial slices agee 30° from the AC–PC plane, no gap, interleaved. rsfMRI abstracts were calm and followed by bristles anatomic runs of aggregate acquisition. During the resting-state abstracts browse which lasted for 6 min 48 sec, participants were instructed to abutting their eyes while lying awake, and not to anticipate of annihilation in accurate or in a analytical way. In anatomic runs, stimuli were projected assimilate a awning at the end of the scanner, and participants beheld stimuli through a mirror army on the arch coil. Participants responded with a magnet-compatible button box. For structural angel acquisition, we acclimated 3D-T1-turbo acreage answer arrangement with the afterward parameters: sagittal accretion with TR = 8.3 ms, TE = 3.3 ms, FOV = 198 × 220 mm2, voxel admeasurement = 0.77 × 0.86 × 1.0 mm3, 216 slices, cast bend = 12°, no gap.
Preprocessing and accepted beeline archetypal were conducted application Statistical Parametric Mapping 8 (SPM8 with MATLAB 2014a, Wellcome Department of Cerebral Neurology, London, U.K.). Slice-timing alteration was conducted by resampling all slices about to the average allotment (i.e., 17th slice) in banausic order. The fMRI abstracts for anniversary accountable was realigned to the aboriginal aggregate for motion alteration and alone abstracts sets with ≤ 2 mm acute displacement during an absolute browse were included in this study. Next, the anatomic images were coregistered to the T1-weighted angel and spatially normalized to the Montreal Acoustic Institute (MNI) arrangement provided with SPM8, again resampled into 3 × 3 × 3 mm admeasurement voxels, followed by spatial cutting application a Gaussian atom with a abounding amplitude at bisected best (FWHM) of 8 mm. A high-pass clarify of 1/128 Hz was acclimated to abolish low-frequency noise, and banausic autocorrelation was done with an AR(1) white babble model.
The accepted beeline archetypal (GLM) included 3 abstracted assignment regressors clay activation during the 0-back, 1-back and 2-back blocks, and these regressors were convolved with approved hemodynamic acknowledgment action (HRF). Six motion ambit acquired during alteration and a affair beggarly regressor were additionally included in the architecture matrix. Three beta images apery BOLD action during anniversary action were generated for anniversary alone participant, and these images were entered into the group-level analysis. For the 2nd akin analysis, 2(groups) x 3(conditions) abounding factorial ANOVA were conducted. The between-group agency consisted of “postpartum” and “control”, and the within-group agency consisted of “0-back”, “1-back” and “2-back”. All statistical analyses were adapted for assorted comparisons based on Monte Carlo simulation agnate to an alpha akin of P < 0.0538.
An absolute resting-state appraisal was conducted to investigate built-in alterations of the postpartum accumulation which were not afflicted by task-related activity. We conducted amplitude of low abundance fluctuations (ALFF) and apportioned ALFF (fALFF) analyses to appraise amplitude of built-in academician activity, and bounded accord (ReHo) appraisal to appraise bounded connectivity of built-in academician action application the toolboxes of Abstracts Processing Assistant for Resting-state fMRI (DPARSF adaptation 2.3, www.restfmri.net). ALFF/fALFF analyses approved low abundance amplitudes which reflect ad-lib activation during the comatose state39. In these analyses, abstracts were preprocessed in an identical address to that of the antecedent GLM appraisal except that nuisance corruption was excluded, and added linear-trend abatement and band-pass clarification (0.01–0.08 Hz) were conducted. Then, the time alternation for anniversary voxel was adapted to the abundance area application a Fast Fourier Transform (FFT). The aboveboard basis of the ability spectrum was affected and averaged beyond the low abundance ambit (0.01–0.08 Hz), acquiescent an ALFF amount for anniversary voxel. fALFF was acquired by accretion the atom of ALFF over the accomplished apparent abundance ambit for the accustomed signal. The ALFF and fALFF maps of anniversary actor were normalized to conduct the group-level two-sample t-test. ReHo appraisal estimated voxel-based synchrony of time-series aural a anatomic array during resting-state, which accustomed us to appraise bounded functionality. The abstracts were preprocessed with the aforementioned methods acclimated in the ALFF/fALFF analysis, but cutting was not performed in ReHo appraisal to abstain inflating the results40. Then, Kendall’s accessory of acceding (KCC) which represents bounded affinity was affected on the time alternation of a accustomed voxel with those of its abutting 26 neighbors in a voxel-wise manner41. The consistent ReHo maps were additionally normalized to abate airheadedness beyond participants and underwent a two-sample t-test.
We acclimated these three altered resting-state analyses to analyze neural alterations consistently begin in both amplitude and connectivity aspects in the postpartum accumulation during resting-state. Details for the resting-state appraisal are declared in the Supplementary information.
The DMN and TPN bulge alternative was performed in the rsfMRI. First, the ahead appear afterwards cingulate case (PCC) berry [−2 −51 27] (Talairach coordinates) was called as a priori ROI to accomplish anatomic connectivity maps42. The alternation accessory map was complete for anniversary actor by artful alternation amid the time alternation of the 8 mm apple PCC affectation and that of anniversary actual voxel in the accomplished brain. The z-transformed anatomic connectivity (FC) maps were again averaged to accomplish a group-level DMN map. As the TPN is authentic as regions assuming anti-correlation with the DMN12, −1 abounding FC maps were acclimated to actualize a TPN map. The nodes aural anniversary arrangement were created as abundantly ample clusters (K > 68 voxels) afterwards thresholding the DMN and TPN maps at Z > 0.35. Three clusters amid in the cerebellum arena were afar from the DMN nodes, as ample portions of the created clusters were positioned alfresco the brain. Six generated DMN nodes were amid in the centermost prefrontal case (mPFC), PCC, mutual centermost banausic gyrus (lMTG, rMTG) and mutual angular gyrus (lAG, rAG), and seven TPN nodes were amid in the after antecedent cingulate case (dACC), mutual dorsolateral prefrontal case (ldlPFC, rdlPFC), mutual antecedent insula (laIns, raIns), and mutual supramarginal gyrus (lSMG, rSMG) (Fig. 6).
Six DMN and seven TPN nodes for the arrangement connectivity appraisal were acquired from the resting-state anatomic connectivity analysis. Abbreviations: ACC, antecedent cingulate cortex; lAG, rAG, mutual angular gyrus; laIns, raIns, mutual antecedent insula; ldlPFC, rdlPFC, mutual dorsolateral prefrontal cortex; lMTG, rMTG, mutual centermost banausic gyrus; lSMG, rSMG, mutual supramarginal gyrus mPFC, centermost prefrontal cortex; PCC, afterwards cingulate cortex.
The time alternation of anniversary of the 13 nodes were extracted from preprocessed alive anamnesis assignment images (260 TR) for anniversary participant. The time alternation of anniversary voxel was detrended and normalized, and again averaged beyond voxels, acquiescent the time alternation of anniversary node. Pair-wise cantankerous alternation coefficients were affected amid anniversary brace of the nodes by administering a fractional correlation, while authoritative for motion artifacts, and the all-around beggarly and alive anamnesis amount effect. Specifically, 3 cerebral amount regressors for anniversary action (0-back, 1-back, 2-back) and 6 motion regressors that were convolved with HRF were regressed out. Then, the alternation coefficients were normalized by Fisher’s Z transform to access normality. For anniversary participant, within-DMN connectivity was adjourned by averaging the alternation coefficients of anniversary brace aural DMN nodes, and within-TPN connectivity was adjourned by averaging the alternation coefficients of anniversary brace aural TPN nodes. Inter-network connectivity was affected by averaging the alternation coefficients of anniversary brace amid the DMN and TPN nodes. In adjustment to analyze within- and inter-network patterns amid the postpartum and ascendancy groups, we agitated out a two-sample t-test application anniversary participant’s within-DMN, within-TPN, and inter-network connections.
To analyze if there was any structural aberration amid the postpartum accumulation and the ascendancy accumulation and to acquisition how the structural alterations were accompanying to anatomic changes, we performed voxel-based morphometry analysis. Voxel-based morphometry appraisal was conducted application SPM 12 (Wellcome Institute of Neurology, University College London, UK, http://www.fil.ion.ucl.ac.uk/spm/). T1-weighted images were anecdotal into gray matter, white matter, and cerebrospinal aqueous by an image-intensity contrast alteration and a unified tissue-segmentation procedure. The anecdotal gray amount images were registered application a customized arrangement by DARTEL algorithm (Wellcome Department of Imaging Neuroscience) and spatially normalized to accepted MNI space. Then, the gray amount images were articulate application Jacobean determinants of the agnate anamorphosis filed for acclimation aggregate changes. The consistent images were smoothed application an 8-mm FWHM Gaussian kernel.
The two-sample t-test was performed on gray amount images for group-level analysis, and we acclimated an uncorrected beginning of P < 0.001, k = 100 for basic purposes.
Normally broadcast analytic abstracts were compared amid the postpartum and ascendancy groups with the two-sample t-test. Other abstracts were analyzed with the Mann-Whitney U-test.
Correlations amid estradiol level, behavioral result, and arrangement connectivity were examined. The Pearson alternation appraisal was conducted if both ethics of the alternation brace were commonly distributed, while the Spearman alternation appraisal was conducted for alternation pairs with non-normal distributions.
Statistical analyses were performed with SPSS, Adaptation 24.0 (IBM, Armonk, New York) and the statistical software R (version 3.5.1; R Foundation for Statistical Computing, Vienna, Austria). A two-tailed P < 0.05 was advised to announce statistical significance.
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