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PLOS One logoLink to PLOS One
. 2021 Jun 14;16(6):e0252622. doi: 10.1371/journal.pone.0252622

The effect of colour on reading performance in children, measured by a sensor hub: From the perspective of gender

Tamara Jakovljević 1,*, Milica M Janković 2, Andrej M Savić 2, Ivan Soldatović 3, Ivan Mačužić 4, Tadeja Jere Jakulin 5, Gregor Papa 6, Vanja Ković 7
Editor: Murugappan M8
PMCID: PMC8202909  PMID: 34125863

Abstract

In recent decades reported findings regarding gender differences in reading achievement, cognitive abilities and maturation process in boys and girls are conflicting. As reading is one of the most important processes in the maturation of an individual, the aim of the study was to better understand gender differences between primary school students. The study evaluates differences in Heart Rate Variability (HRV), Electroencephalography (EEG), Electrodermal Activities (EDA) and eye movement of participants during the reading task. Taking into account that colour may affect reading skills, in that it affects the emotional and physiological state of the body, the research attempts to provide a better understanding of gender differences in reading through examining the effect of colour, as applied to reading content. The physiological responses of 50 children (25 boys and 25 girls) to 12 different background and overlay colours of reading content were measured and summarised during the reading process. Our findings show that boys have shorter reading duration scores and a longer Saccade Count, Saccade Duration Total, and Saccade Duration Average when reading on a coloured background, especially purple, which could be caused by their motivation and by the type of reading task. Also, the boys had higher values for the Delta band and the Whole Range of EEG measurements in comparison to the girls when reading on coloured backgrounds, which could reflect the faster maturation of the girls. Regarding EDA measurements we did not find systematic differences between groups either on white or on coloured/overlay background. We found the most significant differences arose in the HRV parameters, namely (SDNN (ms), STD HR (beats/min), RMSSD (ms), NN50 (beats), pNN50 (%), CVRR) when children read the text on coloured/overlay backgrounds, where the girls showed systematically higher values on HRV measurements in comparison to the boys, mostly with yellow, red, and orange overlay colours.

Introduction

In children reading skills progress over developmental stages [1], while learning to read is one of the most important achievements of the early school years [2]. The complex process of reading skill acquisition involves perception and cognition, via integration of auditory and visual information processing, and memory, attention, and language skills [3]. Reading skills depend upon a range of cognitive abilities and perceptual processes that affect learning during development [4–6]. Gender differences in cognitive abilities during development have been widely analysed in neuropsychological and psychological research. However, these differences are still subject to debate [7–11]. Some studies have reported such differences [12, 13], while others report that they are not able to isolate them [14–18]. This implies the need for additional research on gender differences during cognitive development. More generally, in the past decade, the question of whether males and females differ in cognitive ability has been the focus of significant research [19]. While males and females do not differ in general intelligence, which is a general consensus [20], gender differences are commonly observed for more specific cognitive abilities such as visual-spatial ability [21] and language [22]. However, most gender differences are small or trivial (close to zero) in magnitude, explained by the Hyde gender similarities hypothesis (GSH) [15]. The gender gap in reading achievement, which is found cross-culturally, may be one exception to this hypothesis [19, 23, 24]. Hyde [25] concluded that it is “difficult to reconcile” the magnitude of the gender gap observed in reading with that in other domains of verbal ability, which is typically much smaller, as is claimed by Linn [26].

In the developmental context, girls tend to be superior to boys in verbal abilities and linguistic function from infancy through to adulthood, and for this reason gender difference has been investigated in previous studies, reporting greater reading achievements in girls [24, 27], with boys being better in visuospatial tasks involving memory [28]. Female students read more frequently and had a more positive attitude towards reading, resulting in better reading comprehension [29]. Recent research reports a larger Scan Path Length and Saccade Amplitude in female subjects [30]. Also, in tests of early reading ability Harper and Pelletier [31] found no gender differences in children’s performance. However, the study employing eye-tracing methodology revealed gender differences in reading abilities indicated by Saccade Duration, Regression Rate, and Blink Rate [27].

Currently there are very few sources that explore the impact of colour on the reading process, specifically with early school-age children [32, 33]. The influence of text-, background-, or overlay-colour on the reading process in children is reported in literature [34–36] but there is no clear consensus regarding this. A recent study reported that colour does not influence the reading process [37], while conversely another has found that colour may be particularly effective for early readers such as school-age children [38]. In the study of visual stress, it is found that male subjects prefer blue and green, and females prefer pink and purple overlay colours for reading [39]. Therefore, the effect of colour on the reading process from the perspective of gender is interesting for further investigation.

Reading involves attention, memory, and sensory integration, which may be reflected in the psycho-physiological state of the individual engaged in the reading task. These processes are a result of fundamental physiological and neural processes, which are measurable by different BioSignal modalities such as Electrocardiography (ECG), Electroencephalography (EEG), Electrodermal Activity (EDA), and eye movement. The goal of the recent study was to incorporate multimodal sensor measurements to investigate the effect of colour on the content within the reading task in children from the perspective of gender differences. We have included measurements of heart rate variability (HRV), EEG, EDA, and eye-tracking to assess the influence of background and overlay colour on reading performance in boys and girls attending the early years of primary school. We aimed to address the mechanisms of colour effect on the reading process through electrophysiological correlates of the reader’s state while taking into account the gender aspect of the reading acquisition.

The current research is an attempt to present new evidence regarding gender differences in reading skill. This research aims to contribute to the existing body of knowledge on the effect of the text, background and overlay colour according to gender.

We aimed to investigate the effects of colour on the content as a stressor during the reading task. The present study aims to further illuminate underlying physiological and behavioural processes accompanying the reading task in children from the gender perspective.

Materials and methods

Participants

The study was carried out with fifty healthy participants, boys and girls (25 plus 25) randomly chosen from students in the second and third years (aged 8–10) of primary school “Drinka Pavlović” in Belgrade. Inclusion criteria were that children have normal or corrected-to-normal vision, and no reading and learning disabilities or attention disorders. A typical case for exclusion would be the presence of large artefacts in the acquired signals. However, no such cases were observed in our sample. According to these criteria, no participants were excluded from the statistical analysis.

Children individually participated in the experiment, each under the same experimental conditions: during the school day and in the same small classroom. They received a short instruction about the experiment setup. After they finished the reading test, participants received a certificate and a small present from the researcher. The research process was anonymous and the collected data were anonymised. Only the data regarding the gender and age of the participants were available to the research team. Before starting the experiment, researchers received oral informed consent about the students’ participation from the parents at a school class meeting, organised by the school director and the class teacher, which was summarized in the school director’s note and delivered to researchers.

The ethical committee of the Psychology Department of the University of Niš (a branch of the Serbian Psychology Association) approved the experimental procedure No 9/2019.

Procedure

A computer screen and keyboard were placed in front of every child. The participants read the text in silence from the computer screen, as per the instructions from the researcher at the beginning of the experiment. Each of the participants read the story from the stimuli presentation on the screen, pressing the space button to receive the next paragraph on the next slide. The experiment started with the presentation of black text on white background (the referent slide) as children would typically see in daily life. After that, a pseudo-randomised background colour with black text was presented to the children along with an overlay version (marked by O in the further text, e.g. “red O is for red overlay”) of the presented slides. The text on each colour/slide was different but was kept in a logical order. Except for the referent slide, no other colour was fixed to any particular stimuli presentation. There are no other factors (semantic or affective content, syntax, vocabulary, or text complexity) that could impact the reading process apart from the actual colour, given that the colours were randomly presented to the participants, rather than being selected.

The experiment design was exactly the same as in [40].

Fig 1 shows the sensor hub which consists of a portable multimodal ECG/EEG/EDA and eye tracking system for physiological data acquisition during the reading task. For data real-time monitoring and storage two laptops were used, one for the ECG/EEG/EDA system, and one for eye movement signal monitoring (connected with an external keyboard and the screen positioned in front of the child).

Fig 1. Portable multimodal ECG/EEG/EDA system and eye-tracking system.

Fig 1

A portable remote eye tracker (SMI RED-m 120-Hz, https://www.smivision.com) was mounted in front of the participants and fixed to the screen to secure its stability. In order to ensure that each participant was the same distance from the screen, an adjustable chin-rest was mounted on the table (it was placed 16 cm above the table and 57 cm from the eye-tracking sensor) [41]. For the stimuli presentation SMI Experiment Centre 3.7 was used. An iView RED-m was used for data storage and collection.

The Smarting (mBrainTrain, Belgrade, Serbia) mobile system with a 24-channel EEG amplifier was used for recording EEG and ECG signals, which were communicating wirelessly with a laptop via Bluetooth. In the experiment a Greentek Gelfree-S3 cap with twenty-two monopolar EEG channels was used (10/20 locations: Fp1, Fp2, F3, F4, C3, C4, P3, P4, O1, O2, F7, F8, T7, T8, P7, P8, Fz, Cz, Pz, AFz, CPz, POz). The FPz electrode was used as the ground site and for the reference site the FCz electrode was used. The ECG signal was recorded by one channel of the Smarting amplifier using a surface SKINTACT ECG electrode placed at the left chest region above the heart. The signals of EEG and ECG were acquired with 250 Hz sampling rate and 24-bit resolution. Prior to the test, the skin-electrode impendence was below the manufacturer’s recommended value of 1 kOhm.

For the synchronisation of the multimodal ECG/EEG/EDA and the eye tracking system, one channel of the Smarting amplifier with a small photosensitive sensor was used. This sensor registered the colour changes between two subsequent slides (one black and one white slide lasting for 200 ms each, positioned between two presented slides) indicating the colour changes of the presented slides.

A custom-made galvanic skin response device [41] that sends data via Bluetooth to a laptop was used (sampling rate 40 Hz) for electrodermal activity (EDA) acquisition. EDA data were recorded on the laptop using the Smarting application.

Data processing

BeGaze 3.7 software was used to monitor eye tracking data. Eye tracking analysis included the following parameters: a) Fixation Count, b) Fixation Frequency (count/second), c) Fixation Duration Total (ms), d) Fixation Duration Average (ms), e) Saccade Count, f) Saccade Frequency (count/second), g) Saccade Duration Total (ms) and i) Saccade Duration Average (ms).

EEG/ECG/EDA data were analysed using Matlab ver. 8.5 (Mathworks, USA) in the following manner for each presented slide:

  1. EEG data of all subjects was processed offline. EEG signals were band-pass filtered using 4th order Butterworth filter to extract the activity in the following frequency bands: Delta (0.5–4 Hz), b) Theta (4–7 Hz), c) Alpha (7–13 Hz), d) Beta (15–40 Hz) and e) broadband EEG activity (0.5–40 Hz).

    Filtered signals of all subjects/channels were squared and segmented according to the event markers while each epoch was associated with the reading task of one slide. The median value of data associated with each epoch was calculated for obtaining a single band-power value. Median calculation is used to remove impulse-noises associated with movements, blinks and other artefacts that may occur in the EEG during reading within each epoch. Additional visual inspection of power epochs was conducted to ensure that the median values represent the valid quantification of the band-power activity of each epoch.

  2. Heart activity beats were extracted using Kubios HRV Premium 3.3.1. software [42, 43]. The beats are detected using the Kubios built-in algorithm based on the Pan–Tompkins algorithm [44]. The period between two beats, so called beat-to-beat interval (BBI), and time domain heart rate variability (HRV) parameters [45], Table 1, were extracted by the same software. Also, the Kubios built-in threshold based artefact correction algorithm was performed (a local average interval of 0.35 s was selected and the detected artefacts were automatically replaced by cubic spline interpolated values within the software).

  3. The average value of EDA data was calculated for each slide.

Table 1. HRV parameters.

Parameter (Unit) / Time domain parameters Description
    Mean RR (ms) Mean value of BBIs
    SDNN (ms) Standard deviation of normal BBIs
    Mean HR (beats/min) Mean value of heart rate
    STD HR (beats/min) Standard deviation of heart rate
    CVRR = SDNN/Mean RR (n.u.) Coefficient of variance of normal BBIs
    RMSSD (ms) Root mean square of differences of successive BBIs
    NN50 (beats) Number of successive BBIs that varied more than 50 ms
    pPNN50 (%) Percentage of successive BBIs that differ more than 50 ms

Statistical methodology

Here we present results as percentages, means ± standard deviation or taking into account data type and distribution. We compared groups (boys vs. girls) using a parametric test, an independent samples t-test. All p-values which were less than 0.05 were considered significant. The data were analysed within the SPSS 20.0 software (IBM Corp. Released 2011. IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.). The Bonferroni corrections were applied in all the statistical analysis where necessary as a control for multiple comparisons.

Results

Reading results on white (default) background with black text

Gender comparisons (girl vs. boys) regarding the examined parameters for white background only are presented in Table 2. A significant difference has been obtained regarding a single HRV parameter for pNN50 (%), where girls have higher scores in comparison to boys. In all other parameters of EEG frequency bands (Alpha, Beta, Theta, Delta), ECG parameters and Eye tracking measurements, we observed no significant difference between girls and boys.

Table 2. Reading duration, EEG, eye tracking, EDA and HRV parameters in girls and boys—significant p values are marked as bold.

Parameters Grade p value*
MALE (n = 25) FEMALE (n = 25)
Reading duration
    RD (s) 40.32± 21.64 49.04± 23.49 0. 21
EEG parameters (median power band)
    Alpha 12.64± 8.47 11.99± 6.56 0.76
    Beta 5.50± 3.00 5.77± 2.89 0.75
    Delta 133.64± 198.80 81.32± 52.95 0.21
    Theta 20.31± 28.69 16.19± 9.54 0.50
    Whole Range 134.57± 76.02 130.77± 76.62 0.86
Eye tracking parameters
    Fixation Count 39.88± 21.48 37.68± 15.40 0.69
    Fixation Frequency [count/s] 1.02± 0.45 0.97± 0.52 0.74
    Fixation Duration Total [s] 48.29± 47.98 44.30±22.18 0.72
    Fixation Duration Average [ms] 1,120.83± 588.25 1,201.85± 554.00 0.63
    Saccade Count 34.68± 11.23 32.32± 14.59 0.53
    Saccade Frequency [count/s] 0.95± 0.45 0.86± 0.52 0.53
    Saccade Duration Total [ms] 784.50±289.72 722.84±361.63 0.52
    Saccade Duration Average [ms] 22.49± 3.75 22.38± 5.59 0.94
EDA value
    EDA (uS) 7.66± 3.71 7.69± 3.61 0.98
HRV parameters
    Mean RR (ms) 664.09± 54.58 673.95± 99.37 0.67
    SDNN (ms) 40.37± 19.12 54.34± 36.03 0.09
    CVRR (n.u.) 0.07±0.03 0.08±0.04 0.20
    Mean HR (beats/min) 90.93± 7.43 90.76± 12.36 0.95
    STD HR (beats/min) 5.51± 2.35 6.89± 3.09 0.08
    RMSSD (ms) 48.75± 29.40 69.97± 56.33 0.10
    NN50 (beats) 11.60±11.75 20.32±17.63 0.05
    pNN50 (%) 22.76±17.75 36.78±25.63 0.03

Independent sample t test

Background and overlay colours

In Table 3 a comparison between girls and boys, based on the t-test for independent samples, was obtained on each of the parameters measured in the study, namely: Reading duration, EEG, Eye tracking, EDA and HRV. As is obvious from the table, the girls (coloured in red) scored systematically higher in many of the HRV measurements. In particular for SDNN (ms) they scored higher on yellow, red O, orange O, and purple O; for CVRR they scored higher on yellow, red O, yellow O, orange O and purple O; for STD HR girls scored higher on red, yellow, orange, turquoise, red O, blue O, yellow O, orange O and purple O; for RMSSD they scored higher on yellow, turquoise, red O, yellow O, orange O and purple O; for NN50 they scored higher on red, yellow, orange, turquoise, red O, blue O, yellow O, orange O and turquoise O; and for pNN50 they scored higher on yellow, orange, purple, turquoise, red O, yellow O, orange O and purple O. Boys only scored higher when reading on a purple background for the following eye-tracking parameters: Saccade Count, Saccade Duration Total and Saccade Duration Average (coloured in blue).

Table 3. Differences between girls (marked with red colour) and boys (marked with blue colour) on reading duration, EEG, eye tracking, EDA and HRV parameters (p < .05).

 Parameters Normalized values
red blue yellow orange purple turquoise red O blue O yellow O orange O purple O turquoise O
Reading duration
    RD (s)
EEG parameters (median power band)
    Alpha
    Beta
    Delta
    Theta
    Whole Range
Eye tracking parameters
    Fixation Count
    Fixation Frequency [count/s]
    Fixation Duration Total [s]
    Fixation Duration Average [ms]
    Saccade Count
    Saccade Frequency [count/s]
    Saccade Duration Total [ms]
    Saccade Duration Average [ms]
EDA value
EDA (uS)
HRV parameters
    Mean RR (ms)
    SDNN (ms)
    CVRR
    Mean HR (beats/min)
    STD HR (beats/min)
    RMSSD (ms)
    NN50 (beats)
    pNN50 (%)

Girls vs. boys across all of the examined parameters over averaged scores aggregated for all tested colours are presented in Table 4. Boys achieved higher scores on a few EEG and eye-tracking measurements, namely: Delta and Whole range EEG band measurements and Fixation Count, Saccade Count and Saccade Duration Total. The girls, on the other hand, scored higher on the Reading Duration and on a few HRV measures, namely: SDNN, STDHR, RMSSD, NN50, PNN50 and CVRR.

Table 4. Reading duration, EEG, eye tracking and EDA parameters in girls and boys across all colours together—significant p values are marked in bold.

Parameters Grade p value*
MALE (n = 25) FEMALE (n = 25)
Reading duration
    RD (s) 41.83± 22.61 48.83± 27.72 0.00
EEG parameters (median power band)
    Alpha 11.02± 6.06 10.50± 5.94 0.27
    Beta 5.32± 2.61 5.50± 3.82 0.50
    Delta 82.90± 81.16 61.04± 38.66 0.00
    Theta 15.31± 14.16 13.98± 8.35 0.14
    Whole Range 116.90± 69.03 103.82± 58.33 0.01
Eye tracking parameters
    Fixation Count 39.55± 22.15 35.79± 11.74 0.01
    Fixation Frequency [count/s] 1.01± 0.48 1.00± 0.82 0.81
    Fixation Duration Total [s] 47.49± 39.36 45.22±26.61 0.41
    Fixation Duration Average [ms] 1,154.48± 534.75 1,196.31± 554.87 0.34
    Saccade Count 34.95± 15.10 30.75± 10.01 0.00
    Saccade Frequency [count/s] 0.93± 0.46 0.88± 0.87 0.33
    Saccade Duration Total [ms] 776.14±441.89 667.28±246.73 0.00
    Saccade Duration Average [ms] 21.76± 3.27 22.01± 5.27 0.48
EDA value
    EDA (uS) 7.56± 3.24 7.83± 3.64 0.31
HRV parameters
    Mean RR (ms) 656.20± 51.22 662.65± 90.05 0.26
    SDNN (ms) 40.08± 16.87 52.59± 29.66 0.00
    Mean HR (beats/min) 92.00± 7.31 92.10± 11.36 0.89
    STD HR (beats/min) 5.46± 1.99 6.90± 2.47 0.00
    RMSSD (ms) 44.89± 23.32 65.14± 46.05 0.00
    NN50 (beats) 12.25±10.57 21.51±18.19 0.00
    pNN50 (%) 21.45±16.03 34.23±24.02 0.00
    CVRR 0.07±0.03 0.09±0.03 0.00

*independent sample t test

Discussion

The Reading Duration, EEG, eye tracking, EDA and HRV parameters were evaluated in 50 children (25 female and 25 male second and third year students (aged 8–10) of primary school) using a multimodal sensor hub. As reading process involves attention, memory, and sensory integration, which may be reflected in the psychophysiological state of the individual engaged in the reading task, the study aim was investigating different BioSignal modalities such as ECG, EEG, EDA, and eye movement during the reading task.

Gender differences in reading are widely reported [2, 19, 24, 27, 46–49], and it was found that motivation, attitudes, and the type of reading task could impact on reading skills in boys more closely than in girls [47]. It was speculated that the boys’ reading performance could depend more on their motivation and attitude. The results of the present study showed that boys had shorter reading duration parameters than girls, but at the same time, they scored higher in some eye- tracking measures, and had longer Fixation Count, Saccade Count, and Saccade Duration Total measurements than the girls, irrespective of background/overlay colour. They also had a longer Saccade Count, Saccade Count Total and Saccade Count Average when reading text on a purple background. Additionally it is reported that males have a more positive emotional response than females during competitive game play [50]. Therefore, here we have taken into account findings that suggest that boys are more motivated to read in new conditions, without teachers’ grades/assessments, and have more competitive attitudes in comparison to female students. It has also been reported that male students have poor reading abilities in comparison to girls [27, 51], and therefore make more exploratory eye movements, which consequently result in larger Saccade Amplitudes as we have also demonstrated in the present research.

Regarding the normal maturation processes reflected in the EEG, McCarthy reported [52] that gender differences are equally distributed, while other researchers [53, 54] found that EEG differences between boys and girls suggest earlier maturation in girls [55]. On the other hand, Cohn [56] and Gasser [57] found no differences between boys and girls measurable by EEG. It is also reported that the amount of activity in the lower frequency EEG bands decreases with age, and in higher frequency bands, it increases [58, 59]. The gender differences in EEG are also reported in context of the task performance and cognitive activity. During the task and rest phase, females have a higher EEG power than males [60, 61]. Also, gender differences are frequently reflected in numerous factors, like task and age [62]. Some research shows that in most cognitive tasks including language, there exist inappreciable differences in behavioural output between the genders [16–18]. EEG power and its distribution over a lifetime also varies between the genders [63]. EEG maturation markers increase in faster band activity (alpha, beta) and decrease in slower band activity (theta, delta) [55, 57, 64–66]. In the present study it is shown that boys have higher values of delta band and whole range of EEG in comparison to girls. Clarke et al. report that boys’ EEG matures faster than girls’ in childhood, but that these differences are eliminated during adolescence. Conversely Gasser et al. [57] declare that there are no gender differences, mostly because of high interindividual variability in the EEG power spectrum. Other authors have found that until the age of 16 the alpha rhythm does not mature [67]. In previous research the delta band was found to be higher in young individuals than in adults because of incomplete cortical maturation, and is typically even higher in children with learning disabilities [68]. These findings are in line with our results, which showed increased Delta and Whole Range EEG bands to be more prominent in boys than in girls, which could be as a result of the faster maturation process in girls. This is in keeping with previously mentioned results showing that boys had a less mature pattern of eye-movements in comparison to girls.

Likewise, electrodermal activity has been used in several studies with the objective of clarifying markers of psychophysiological functioning and children’s developmental processes [69, 70].

Several studies supported higher levels of baseline SCL (Skin Conductance Level) [68] and SCL reactivity to stressors [71] in girls in comparison to boys. In the present study we found no evidence of systematic differences between boys and girls for this measurement when reading text on either white or coloured/overlay background.

Physiological mechanisms during adolescence actively and progressively undergo changes. It has been reported that HRV progressively reduces with age, and development during adolescence can be assessed using the heart rate variability (HRV) [72]. HRV can be used to ascertain the evolution of the ontogenetic maturation [73–76]. Moreover, the gender influence measured by HRV parameters was manifested only in young adults and younger adolescents and our study group belongs to the same age category. Research study shows that measurements of HRV depend on the age but not on the gender of healthy children [77]. When different colours of background, text, and overlay were included in the reading process we found significant differences between girls and boys, whereby girls scored higher on HRV parameters. This indicates higher emotional reactions in girls when they read the text on the coloured/overlay background in comparison to the boys. Regarding this result, our findings are compatible with previously reported results showing that girls have higher values on SDNN and RMSSD measurements.

Conclusion

Primarily this research aimed to assess gender differences in the reading process and to contribute to existing research on the effect of text, background and overlay colour according to gender.

Secondly, the aim was also to investigate the effects of colour on the content as a stressor in the reading task. In order to shed light on contradictory reports regarding gender differences in reading skills, present study illuminates underlying physiological and behavioural processes in the reading task in children from the gender perspective. It evaluates differences in reading duration, EEG, ECG, EDA and eye movement measures on both white and 12 different background/text/overlay colours. It was found that boys show shorter reading duration parameters than girls, and at the same time longer eye-tracking measures such as Fixation Count, Saccade Count, and Saccade Duration Total while reading on a coloured background/overlay, whereas they had a Longer Saccade Duration, Saccade Duration Total, and Saccade Duration Average on a purple background. These results partially support our expectation that boys would have more difficulties reading the text when displayed on background/overlay colours. However, they did not have more issues reading on the coloured background in comparison to the white/default background.

Comparing EEG parameters in girls and boys during reading on white background we did not find systematic differences. Observing all the colours together, it is shown that boys have higher values in Delta and Whole Range bands in comparison to the girls. As the Delta Range is higher in young adults, the findings are aligned with previous research where it is shown that boys will have more difficulties in reading tasks because the reading process is still not automated in comparison to the girls. In fact, they have also demonstrated longer Saccade Count and Saccade Duration measurements in comparison to the girls when reading on a purple background. It seems that the colour can really increase the task difficulty for less proficient readers. We did not find systematic differences for EDA measures between boys and girls while reading on white or coloured background/overlay content. However, regarding ECG measures, girls scored significantly higher on HRV measures (SDNN (ms), STD HR (beats/min), RMSSD (ms), NN50 (beats), pNN50 (%), CVRR), in particular on yellow, orange O and red O colours. These findings are also contributable to studies where it is shown that girls have higher values on HRV measures than boys, which is particularly evident from results including the additional effect of colour on the reading process.

Finally, we can underline that colours used as a stressor in a particular reading task could illuminate gender differences, especially in eye-tracking and ECG measures. Boys have shown longer Saccade Count and Saccade Duration in comparison to girls while reading on the purple colour. Boys have shown shorter reading duration than girls on all coloured background/overlay, and longer eye-tracking measures such as Fixation Count, Saccade Count, and Saccade Duration Total. Regarding the ECG (SDNN, STD HR, RMSSD, NN50, pNN50, CVRR) measures, girls scored higher than boys while reading on yellow, orange O, and red O colours. These findings show that colours could be contributing to a better understanding of gender differences and their relation to the context of the reading processes.

Supporting information

S1 Data

(XLSX)

Acknowledgments

The authors acknowledge Elementary School “Drinka Pavlović” (Belgrade) and IPS Jozef Stefan.

Data Availability

All relevant data are within the manuscript and S1 Data.

Funding Statement

The Authors work were supported through the Slovenian Research Agency (research core funding No. P2-0098), AD Futura Fund (Public Scholarship, Development, Disability and Maintenance Fund of the Republic of Slovenia, and the Ministry of Education, Science and Technological Development of the Republic of Serbia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Decision Letter 0

Murugappan M

23 Feb 2021

PONE-D-20-40129

Effect of colours on reading performance in children measured by the sensor hub: from the perspective of gender

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Partly

Reviewer #3: No

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Comments to the author:

This study aims to investigate the gender differences in reading achievement, cognitive abilities and maturation process using HRV, EEG, and EDA. The study is interesting, and the paper is well-written. There are however some major areas that require the authors' attention.

My major concern is about processing of bio signals. Generally, the physiological signals are affected by noise (e.g., power line interference) and artifacts (body movements). Especially, when you record signals from children. I was unbale to find the details about signal pre-processing. How the noise and artifacts are removed? How the authors extracted the band power? What filter used to extract the EEG sub-frequency bands? Overall, no clear information about pre-processing.

Next, I was unable to see the information about data analysis i.e., data segmentation or the authors analyzed the whole recorded signals. state the outcome of this research in real time? What is the significant of this research? Include the limitations

Reviewer #2: The research article analyzes various physiological and eye gaze parameters to understand the gender differences in reading comprehension of elementary students. The differences is observed in many of the parameters and concurs with existing research. Colors do not seem to influence children with good proficiency.

Some comments

1. What is the novelty proposed in this research work? How are the differences in controversial reports filled in this work?

2. Studies on the influence of color on reading can be understood by different questionnaire based methods and statistical analysis of the same. Was any such study done or feedback obtained from the teachers to validate your results. Is there a need to use invasive methods such as EEG?

3. Why do you think that the delta waves show significant differences?

4. What would the proposed application of these finding. Proposing a few may provide more insight into the paper.

Reviewer #3: The problems statement is good. Some of the results are presented but the signal processing aspects are missing.

Major comments

1. The paper is poorly written. Please check typos and rewrite the paper in standard English

2. Signal processing techniques are missing in this paper. It should be described clearly with mathematical expressions, algorithms with optimal coding parameters and also results.

**********

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: Yes: M Sabarimalai Manikandan

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PLoS One. 2021 Jun 14;16(6):e0252622. doi: 10.1371/journal.pone.0252622.r002

Author response to Decision Letter 0


22 Apr 2021

Dear Reviewers, 

Thank you for your detailed and useful comments. We hope that you will find our answers satisfactory and we definitely feel that your comments helped us improve the manuscript significantly. 

Sincerely, 

Authors of the manuscript

Reviewer #1: Comments to the author:

This study aims to investigate the gender differences in reading achievement, cognitive abilities and maturation process using HRV, EEG, and EDA. The study is interesting, and the paper is well-written. There are however some major areas that require the authors' attention.

My major concern is about processing of bio signals. Generally, the physiological signals are affected by noise (e.g., power line interference) and artifacts (body movements). Especially, when you record signals from children. I was unabale to find the details about signal pre-processing. How the noise and artifacts are removed? How the authors extracted the band power? What filter used to extract the EEG sub-frequency bands? Overall, no clear information about pre-processing.

Next, I was unable to see the information about data analysis i.e., data segmentation or the authors analyzed the whole recorded signals. state the outcome of this research in real time?

Response:

We have included more detailed information about signal processing conducted within this study, such as type of filter, calculation of band-power, signal segmentation and inspection for artifacts and their removal. Fourth order Butterworth filter was used to extract the activity of 5 frequency ranges analyzed in this study. Median values of band-power were calculated for each epoch (slide)/ frequency band in order to obtain a single power value for the total reading duration of each slide. The choice of median power calculation was introduced in order to remove impulse-noises associated with movements, blinks and other artefacts which was stated in the text. Additional visual inspection of band-power values for each subject/band/epoch was conducted in order to validate the obtained median values and check for presence of artifacts.

Our study is of exploratory nature and includes offline data processing only for examining the gender differences during reading task (with different colour/overlay setups) using multimodal signal measurements. Future studies and analyses of the collected data may include exploration of optimal signal measurement setup which could in real time estimate the preferred colour/overlay in order to facilitate the reading task, but this is out of the scope of the current study. You can find more in Methods section (Page 5).

What is the significant of this research? Include the limitation

Response:

Regarding the significance of this research -  the second reviewer in his first question also asked about it and about the novelty of the results, so hopefully you will find our answer satisfying.

One of the limitations was that we could have developed a longitudinal study where we would follow a group of girls and boys for a number of years, but this was outside of the scope of the current study and exceeds our current resources.

Reviewer #2: The research article analyzes various physiological and eye gaze parameters to understand the gender differences in reading comprehension of elementary students. The differences is observed in many of the parameters and concurs with existing research. Colors do not seem to influence children with good proficiency.

Some comments

1. What is the novelty proposed in this research work? How are the differences in controversial reports filled in this work?

Response:

The novelty and significance of this research is primarily methodological - we attempted to develop a sensory hub in order to simultaneously test cognitive and emotional arousal in boys and girls whilst reading on different colour/overlay backgrounds in order to overcome a pure behavioural measurements and get a more fine-grained insight into the processes and differences involved in process of reading across the two groups. The differences observed in controversial reports may have to do with strategic responding to some extent (given that the behavioral responding does not capture the underlying differences and strategies in the task), which we hopefully overcame by applying an automated way of measuring both cognitive and emotional responses.

2. Studies on the influence of color on reading can be understood by different questionnaire based methods and statistical analysis of the same. Was any such study done or feedback obtained from the teachers to validate your results. Is there a need to use invasive methods such as EEG?

Response:

Yes, we actually informally interviewed teachers as well as some speech specialists in the field regarding reading on the coloured background/overlay backgrounds, and they suggested that this kind of intervention may be of a great help in focusing attention in children whose attention span is much shorter nowadays. We did not run a separate study based on the feedback, but it become clear to us that, apart from looking into group differences, like in this study, the future research will need to focus more on the individual differences, as it seems that kids tend to have their own prefered colour and that choice vary significantly. But, that would be a matter of some future research, as it exceeds the scope of the article and results we presented here. 

We ensure you, as we did teachers and parents of the children who took part in this study, that none of the methodologies we used here is invasive to children in any way. We literally told them that it is measuring brain waves and not changing them, in the same way as we can measure body temperature without changing it. The amount of infra-red light used in the eye-tracking system is definitely such that it can not cause any harm to childrens’ eyes. Otherwise, we, as researchers, would have a huge ethical dilemma, even before getting a formal ethical approval for this study (which we did). Also, kids felt that they took part in a scientific adventure and they loved it - to the extent that they would come back to the researchers during the break between classes. 

3. Why do you think that the delta waves show significant differences?

Response:

This is a really interesting question. In our opinion the delta waves show significant differences because boys get to mature later in comparison to girls, and previous research (55, 70) demonstrated that younger (less matured children and children with learning disabilities)  tend to have more prominent delta waves. In our research we also found that boys had a less mature pattern of eye-movements as we interpreted/described in the discussion.  

4. What would the proposed application of these finding. Proposing a few may provide more insight into the paper.

Response:

One would be that we can make the reading process easier by selecting and adjusting the colour of the background for each individual child. The second would be that by applying this system we could help prevention, but also early detection of potential problems that children may have with reading. The major plan would be to extend this stream of research to dyslexic children and to employ machine learning in order to be able to do clear adjustment for each individual child.

Reviewer #3: The problems statement is good. Some of the results are presented but the signal processing aspects are missing.

Major comments

1. The paper is poorly written. Please check typos and rewrite the paper in standard English

Response:

Thank you so much - you were absolutely right. We asked a professional and also a native speaker of English to do the necessary corrections and there were so many of them that we did not keep track-changes of them, because the text would be difficult to read and major corrections difficult to spot. We also intend to get our language expert to make a final read before the paper gets published, if you find all the changes we made satisfactory. 

2. Signal processing techniques are missing in this paper. It should be described clearly with mathematical expressions, algorithms with optimal coding parameters and also results.

Response:

We have used the Kubios HRV Premium 3.3.1software for the heart beat extraction and beat-to-beat interval calculation, as well as for time domain HRV parameters calculation. We have properly referenced this software (added two references: [42], [43]) and we have modified the corresponding paragraph in the text (Page 5):

“2) heart activity beats were extracted using Kubios HRV Premium 3.3.1. software [42, 43]. The beats are detected using the Kubios built-in algorithm based on the Pan–Tompkins algorithm [44]. The period between two beats, so called beat-to-beat interval (BBI), and time domain heart rate variability (HRV) parameters [45], Table 1, were extracted by the same software. Also, the Kubios built-in threshold based artefact correction algorithm was performed (for local average interval was selected 0.35 s and the detected artefacts were automatically replaced by cubic spline interpolated values within the software).”

[42] https://www.kubios.com/hrv-premium/ 

[43] Tarvainen, M. P., Niskanen, J. P., Lipponen, J. A., Ranta-Aho, P. O., & Karjalainen, P. A. (2014). Kubios HRV–heart rate variability analysis software. Computer methods and programs in biomedicine, 113(1), 210-220.

Attachment

Submitted filename: Response to Reviewers_Plos One.docx

Decision Letter 1

Murugappan M

19 May 2021

The effect of colour on reading performance in children, measured by a sensor hub: from the perspective of gender

PONE-D-20-40129R1

Dear Dr. Jakovljevic,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Murugappan M, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

I may strongly suggest the authors to address the reviewer 3 suggestions in the camera-ready version of the manuscript.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Partly

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors addressed all the reviewer comments. I think the paper can be accepted for publication.

Reviewer #2: The authors have addresses the comments. The data collection methods have been justified. The authors have also included signal processing steps for better understanding.

Reviewer #3: Please improve the writing and presentation of this paper.

I would suggest the authors to highlight their statistical findings in the conclusion.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Jerritta Selvaraj

Reviewer #3: No

Acceptance letter

Murugappan M

26 May 2021

PONE-D-20-40129R1

The effect of colour on reading performance in children, measured by a sensor hub: from the perspective of gender

Dear Dr. Jakovljevic:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Murugappan M

Academic Editor

PLOS ONE

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