The aim of this work was to study the quality characteristics, the sensory profiles and the ethylene production of ‘Abate Fetel’ pears sorted at harvest in different TRS maturity classes and stored in different atmospheres. At harvest, 540 pears were measured by TRS for μa670, ranked by μa670 in three maturity classes  and randomized in nine samples according to 1-MCP treatment , storage time (4–6 months) and atmosphere . Fruits were examined at harvest and after 7 days of poststorage shelf life for skin color, firmness, soluble solids, acidity and ethylene production and were submitted to sensory analysis. At harvest and after storage, MoM pears were less green and showed a higher SSC content than LeM ones. After storage, MoM pears produced less ethylene and were perceived to be firmer , more astringent and less juicy (when stored for 6 months) than LeM ones.Time-resolved reflectance spectroscopy (TRS), a nondestructive technique, can help the industry to provide high-quality fruit to encourage pear consumption. The absorption coefficient measured by TRS at 670 nm ( Furthermore, this storage protocol prolonged the storage duration up to 27–35 weeks and, at the same, allowed the fruits to ripen [μa) and the reduced scattering (μs′) coefficients are estimated [μa measured by TRS at 670 nm (μa670) is linked to the biological age of a fruit and represents an effective maturity index for different fruits, such as peaches, nectarines, apples, plums, pears and mangoes [μa670 declines during fruit growth and ripening as well as during storage and shelf life and can be used to segregate fruits into uniform groups based on the maturity degree [μa670, while more mature (MoM) fruits have low values of μa670 [μa670 can distinguish unripe from ripe fruit [The ‘Abate Fetel’ pear, the most important winter pear cultivar in Italy, is appreciated by consumers for its aroma, texture and balanced sweet and sour taste ,2. ‘Abatextended . The 1-mto ripen ,10. A lato ripen ,12. Therto ripen . NIR, Vito ripen . Spectroto ripen ,16,17, tto ripen  or to prto ripen . Howeverto ripen . An imprto ripen . In the to ripen  and measto ripen ,24,25,26stimated ,25. The stimated ,26. The y degree ,29. Fruipe fruit . Similarpe fruit .Thus, the aim of this work was to study the quality characteristics, the sensory profiles and the ethylene production of ‘Abate Fetel’ pears treated with 1-MCP and stored for 4–6 months in different atmospheres in relation to maturity at harvest nondestructively measured by TRS.μa670 at harvest ranged from 0.054 to 1.000 cm−1 and significantly decreased from LeM pears to MoM ones (IAD), texture properties and starch hydrolysis were significantly higher in LeM fruits than in MoM ones, while RSR increased with maturity degree  at harvest to 0.064 ± 0.001 cm−1 after storage plus 7 days of shelf life at 20 °C. The μa670 significantly changed with storage time , 1-MCP treatment , storage atmosphere  and TRS maturity class . The μa670 was significantly higher in 1-MCP-treated pears and in LeM fruits and decreased with storage time only in LeM-1-MCP-treated fruits stored in NA and in CA and in MeM-treated fruits stored in NA , secondly by storage time  and then by TRS maturity class  and day of shelf life . On average, 1-MCP-treated pears developed (mean ± standard error) 47 ± 4 pmol kg−1s−1 and control fruits 417 ± 10 pmol kg−1s−1. Ethylene production was significantly higher in LeM pears (300 ± 21 pmol kg−1s−1) than in MoM ones (223 ± 19 pmol kg−1s−1) and increased from 4 (226 ± 18 pmol kg−1s−1) to 6 months of storage (295 ± 22 pmol kg−1s−1). In control fruits stored for 4 months, ethylene showed the highest production after 3 days of shelf life in MoM pears, while in LeM fruits, ethylene increased from 1 to 3 days, maintaining this level until the end of the shelf-life period , more juicy , sweeter  and aromatic  than 1-MCP-treated ones. The sensory profiles of ‘Abate Fetel’ pears stored for 4 and 6 months in CA and in NA  were maiF-ratio = 7.63, p < 0.01), juiciness  and astringency , with MoM pears being firmer , more astringent and less juicy than LeM ones, especially when stored for 6 months  with pears stored in NA being less sour than those stored in CA , h° (r = 0.42) and firmness (r = 0.44). IAD was positively related to h° (r = 0.58) and firmness (r = 0.48). Firmness, work and stiffness were positively related to sensory firmness, with r ranging from 0.75 to 0.80, and for astringency, r = 0.43, and they were negatively related to juiciness and sweetness, with r ranging from −0.64 to −0.66 and from −0.40 to −0.42, respectively. SSC was positively related to stiffness (r = 0.45) and work (r = 0.40) and to sensory firmness, with r = 0.45. No significant correlation with r ≥ 0.4 was observed for TA. As for sensory attributes, firmness was negatively related to juiciness (r = −0.65) and sweetness (r = −0.43) and positively related to astringency (r = 0.46); juiciness was positively related to sweetness and aroma with r = 0.61 and r = 0.62, respectively; and sweetness was positively related to aroma (r = 0.68). Overall acceptability was positively related to aroma (r = 0.83), sweetness (r = 0.71) and juiciness (r = 0.55).The relationships among variables highlighted by PCA were studied and confirmed by Pearson’s correlation analysis . In factAs for the PC scores, on average, PC1 scores were higher for 1-MCP-treated fruits and for those stored for 4 months, and they did not change with the storage atmosphere or TRS maturity class. PC2 scores were higher for control fruits and for those classified at harvest as MoM by TRS and were not affected by storage conditions. PC1 allowed to discriminate LeM- and MoM-1-MCP-treated pears stored for 4 months (highest PC1 scores) from MoM-1-MCP-treated pears stored for 6 months (intermediate PC1 scores) and LeM-1-MCP-treated pears stored for 6 months (low PC1 scores) and untreated fruits (with any storage time and TRS maturity class), which showed the lowest PC1 scores . PC2 disµa670 decreased from harvest to storage plus 7 days of shelf life at 20 °C, indicating that ‘Abate Fetel’ pears ripen during storage time . In fact, µa670 was higher in 1-MCP-treated fruits that were greener, firmer and contained more acid, and they developed much less ethylene than untreated pears . In the previous works, O2 was set instead at 2%, a percentage that keeps the pears in a less advanced maturity state and inhibits superficial scald but can promote soft scald development [2 were greener and firmer than NA ones after 27 and 35 weeks of storage, while no influence on firmness was observed after 18 weeks. The authors also found that at 18 weeks, CA-stored pears were juicier, more grainy and sourer than NA fruit, while at 27 weeks, they were juicier than CA + NA ones, and after 35 weeks, they were less firm and aromatic than NA ones. The storage atmosphere did not have any significant effect on overall acceptability, no matter the storage time. Also in this case, the different results could be partially explained by the different adopted O2 percentages.The  NA ones , with CA NA ones , even if NA ones . On the elopment ,7. Rizzoµa670 in combination with 1-MCP treatment, as in the control fruits, no change in µa670 was observed compared between 4 and 6 months of storage, while in 1-MCP-treated pears, µa670 decreased with storage time  picked in 2020 at commercial maturity in an orchard of the Protected Geographical Indication (IGP) area located in the Mantova province . At harvest, 540 pears were individually measured on two sides by TRS for the absorption coefficient at 670 nm (μa670). Pears were ranked by decreasing μa670 averaged on the two sides, that is, from the least to the most mature. The ranked fruits were grouped by nines, with a total of 60 groups, corresponding to 60 levels of μa670, and divided into 60 groups, corresponding to 60 μa670 levels. Each fruit from each group was randomly assigned to a different sample. In this way, 9 batches were obtained, each one containing 60 fruits from the whole range of μa670. In each batch, according to fruit ranking, the 60 fruits were divided into three TRS maturity classes: less mature , medium mature  and more mature . The nine batches were randomly assigned to harvest, two 1-MCP treatments , two storage atmospheres  and two storage times (4 and 6 months), as reported in The experiment was carried out on ‘Abate Fetel’ pears  and MoM (rank 51–60) pears were submitted to sensory analysis. At 1, 3 and 6 days of shelf life at 20 °C, ethylene production was measured on LeM (rank 1–5) and Mom (rank 1–5) pears.At harvest, a sample of 60 fruits was analyzed for fruit mass, skin color, flesh firmness, starch hydrolysis, titratable acidity and soluble solids content.An existing instrumentation for TRS developed at the Department of Physics of Politecnico di Milano was used . BrieflyData acquisition was performed by a Time-Correlated Single-Photon Counting board  able to reconstruct the distribution of time-of-flight (DTOF) of the detected photons. The DTOF is analyzed via an in-house-built software based on a nonlinear fitting procedure, thus retrieving the values of absorption and reduced scattering coefficients of the sample. The theoretical curves simulating photon diffusion in a turbid homogeneous medium that are required as input of the fitting procedure are calculated by exploiting a model based on the diffusion equation .h°) was computed according to h° = arctangent(b*/a*) × 360/ (2 × 3.14). Skin color was also measured through the determination of IAD (index of absorbance of chlorophyll) using a DA-meter, a portable nondestructive device based on Vis–NIR spectroscopy . The IAD is the difference between the absorbance values at 670 nm and at 720 nm.Skin color was measured on two opposite sides of the fruit in L*, a*, b* color space using a Spectrophotometer CM-2600d , the primary illuminant D65 and a 10° observer. Hue (−1 to a depth of 8 mm). According to Vanoli et al. [The mechanical properties were determined on two opposite peeled sides of each pear using a penetrometer . Soluble solids content (SSC) and titratable acidity (TA) were determined using juice squeezed from each fruit; SSC was measured by placing some drops of juice on an automatic refractometer ; and TA was determined by titrating 5 g of juice plus 50 mL of distilled water with 0.1 N NaOH to pH 8 using a 682 titroprocessor equipped with a 665 dosimat .Ethylene production was measured according to Rizzolo et al.  on 5 fruSensory analyses were carried out on LeM and MoM pears in a sensory lab equipped with six computerized individual booths, according to Rizzolo et al. . A panelμa670, ethylene production, and quality and sensory parameters were submitted to multifactor ANOVA considering 1-MCP treatment, storage time, storage atmosphere and TRS maturity class as factors; means were compared by Tukey’s test at p ≤ 0.05%. In order to explore the relationship among all variables, all data were submitted to Principal Component Analysis (PCA), and Pearson’s correlation analysis was performed: only significant (p < 0.001) correlations with r ≥ 0.40 were considered. Prior to statistical analyses, the rating scores of each sensory attribute were standardized by panelists according to Bianchi et al. [Data of i et al.  to removµa670) allowed to obtain fruits with different quality and sensory characteristics after storage, mainly regarding skin color, soluble solids content and sensory firmness. However, due to the strong effect of 1-MCP treatment and, secondly, of storage duration, quality characteristics and sensory attributes were not always very different between less and more mature pears. Therefore, TRS maturity classification at harvest can give partial but not conclusive indications of the best storage strategies to obtain fruits that can satisfy consumers.In conclusion, our results indicate that classifying pears at harvest in homogeneous maturity classes by using the absorption coefficient nondestructively measured by TRS at 670 nm (


Humulus lupulus L. (hop) is recognized, but the primary metabolism of the hop strobilus has not been quantified in response to elevated CO2. The photosynthetic contribution of hop strobili to reproductive effort may be important for growth and crop yield. This component could be useful in hop breeding for enhanced performance in response to environmental signals. The objective of this study was to assess strobilus gas exchange, specifically the response to CO2 and light. Hop strobili were measured under controlled environment conditions to assess the organ’s contribution to carbon assimilation and lupulin gland filling during the maturation phase. Leaf defoliation and bract photosynthetic inhibition were deployed to investigate the glandular trichome lupulin carbon source. Strobilus-level physiological response parameters were extrapolated to estimate strobilus-specific carbon budgets under current and future atmospheric CO2 conditions. Under ambient atmospheric CO2, the strobilus carbon balance was 92% autonomous. Estimated strobilus carbon uptake increased by 21% from 415 to 600 µmol mol−1 CO2, 14% from 600 to 900 µmol mol−1, and another 8%, 4%, and 3% from 900 to 1200, 1500, and 1800 µmol mol−1, respectively. We show that photosynthetically active bracts are a major source of carbon assimilation and that leaf defoliation had no effect on lupulin production or strobilus photosynthesis, whereas individual bract photosynthesis was linked to lupulin production. In conclusion, hop strobili can self-generate enough carbon assimilation under elevated CO2 conditions to function autonomously, and strobilus bracts are the primary carbon source for lupulin biosynthesis.The economic value of  Li. Lic appm−2 s−1)  [11]. Lim−2 s−1)  [11]. Dam−2 s−1)  [11]. Th3 leaves  [11].2 of 600 μmol mol−1 increased  via herbicide application. Defoliation did not affect strobilus photosynthetic activity or lupulin production. Blocking bract photosynthesis, to isolate the bract as the sole remaining photosynthetic organ within the strobilus, did prevent lupulin glands from filling. Lupulin glands were amassed approximately equally at the base of each bract in both treatments. Bracts that were not treated yet resided on the same strobilus as some with herbicide treatment filled their basal lupulin glands normally with no visual signs of chlorophyll degradation. Herbicide-treated bracts, however, visually expressed tissue mortality at the site of herbicide contact and failed to fill their lupulin glands . We note2 sources: external atmospheric CO2 and internal CO2 released from organ respiration. Aschan and Pfanz  (An/Ci) and PAR (An/PAR) were repeated four times at ten-day intervals. For the An/Ci curves, strobili were initially acclimated in the chamber until An was stable at a controlled CO2 concentration (415 μmol mol−1) and light level (400 μmol m−2 s−1). The CO2 response curve was constructed at CO2 levels of 415, 300, 200, 150, 100, 50, 25, 415, 600, 800, 1000, 1200, 1500, and 1900 μmol CO2 mol−1 with 10 min of stabilization per CO2 change. After the completion of the An/Ci curve, CO2 concentration was controlled at 415 μmol mol−1 until stabilization, and PAR was sequentially lowered to 1000, 800, 700, 600, 500, 400, 300, 200, 150, 100, 75, 50, 25, and 0 μmol m−2 s−1 with five minutes of stabilization per PAR change.Response curves of net photosynthesis (Aa) to ϕ, as described in Bauerle et al. [2 = 0.968, n = 5; unpublished data).Immediately after gas exchange, five replicate light absorptance samples were recorded per strobilus on random bracts within the strobilus with an SPAD meter and averaged . The SPAD values were used to correct apparent quantum yield  under light saturation due to the capability of estimating in vivo Rubisco activity. The An/Ci data were analyzed as per the fitting method “fitaci” function of the “plantecophys” package [nc), electron transport (Anj), and inorganic phosphate supply (Anp) can limit CO2 assimilation, as summarized in von Caemmerer et al. [The model of Farquhar, von Caemmerer, and Berry  was used package  accordin package , where tr et al. An = min2 in the chloroplast at which photorespiratory CO2 evolution equals the rate of carboxylation  at 25 °C and 21% O2 was as in von Caemmerer et al. [The partial pressure of COr et al. , assuminmax is the light-saturated net photosynthetic rate, ϕa is the apparent quantum yield of assimilation, ø is the convexity of the curve, and Rd is the respiration rate. The parameter ϕa was calculated through linear regression analysis on the initial slope from 20–125 μmol m−2 s−1 to exclude the Kok effect region (≤20 μmol m−2 s−1) [a underestimation from data in the non-linear region (≥125 μmol m−2 s−1) [c) was estimated from the intersection of the regression line with the x-axis (An = 0), and Rd was measured at the end of the An/PAR curve. The quantum yield for CO2 (ϕCO2) was derived from ϕa by correcting ϕa for the percentage leaf absorptance of PAR, as described in Bauerle et al. [n/PAR curves .Photosynthesis versus PAR data were fitted to the nonrectangular hyperbola model of Prioul and Chartier , as descm−2 s−1)  and prevm−2 s−1) . The lige et al. . Equation/Ci parameters were used to parameterize deterministic strobilus-level models for energy balance [2. To place the strobilus CO2 responses in the context of strobilus diurnal carbon gain, the photoperiod was set at 15 h, the approximate day length for strobilus physiological functions in Yakima, WA,  at the beginning of August (day of year 213). Half-hourly averages of incident PAR, temperature, RH, and wind speed in Yakima, WA, were downloaded from the Washington State University AgWeatherNet site. Atmospheric CO2 concentrations of 415, 600, 900, 1200, 1500, and 1800 μmol mol−1 were inputted to estimate strobilus carbon gain responses to CO2 enrichment.Photosynthetic light response and A balance , stomata balance ,49), and balance  (Table 1g) is a direct estimate of carbohydrate biosynthesis:n;light is the total An during the light period, R;dark is the total respiration during the dark period, and 12 is the molecular mass of C. We summed 30 min diurnal carbon estimates to arrive at net daily Cg per strobilus surface area.Predicted strobilus carbon gain  to inhibit electron transport and CO2 assimilation. Per strobilus, several random bracts were swabbed with Paraquat from the bract tip proceeding inward toward the strig, covering approximately two thirds of the top surface of an individual bract. Six replicate plants each per defoliation or control group were treated. Strobili were harvested upon maturity and lupulin gland secretions were visually assessed at 40× magnification on Paraquat-treated and untreated bracts per strobilus.To investigate the carbon source for lupulin biosynthesis on the plant scale, six replicates were randomly assigned to either the “defoliation” or “control” group. The control was left to grow normally, and the defoliation group had all leaves removed using scissors at the strobili stigma senescence stage. The strobili stigma senescence stage, described in the comprehensive strobilus developmental stage index of Kavalier et al. , is infon/PAR and An/Ci curves was six replicates (n = 6), with one strobilus per six replicate plants. Each strobilus was repeatedly measured at ten-day intervals over the course of four repeated measures (40 days). Each plant was an experimental unit treated as a replicate (n = 6). Two-tailed t-tests were used to analyze the significance between each combination of repeated time measurements. Plant response data were analyzed using SPSS ). Differences between means were considered significant when the p value of the t-value was < 0.05.The total sample size for A


Face perception is a fundamental aspect of human social interaction, yet most research on this topic has focused on single modalities and specific aspects of face perception. Here, we present a comprehensive multimodal dataset for examining facial emotion perception and judgment. This dataset includes EEG data from 97 unique neurotypical participants across 8 experiments, fMRI data from 19 neurotypical participants, single-neuron data from 16 neurosurgical patients (22 sessions), eye tracking data from 24 neurotypical participants, behavioral and eye tracking data from 18 participants with ASD and 15 matched controls, and behavioral data from 3 rare patients with focal bilateral amygdala lesions. Notably, participants from all modalities performed the same task. Overall, this multimodal dataset provides a comprehensive exploration of facial emotion perception, emphasizing the importance of integrating multiple modalities to gain a holistic understanding of this complex cognitive process. This dataset serves as a key missing link between human neuroimaging and neurophysiology literature, and facilitates the study of neuropsychiatric populations. For example, the response to fearful faces has been comprehensively delineated using a combination of neuronal firing, local field potentials, and hemodynamic activity in the human amygdala2. Here, we present a multimodal dataset for studying emotion perception and judgment. A key strength of our dataset is that all participants performed the same emotion judgment task  with a gradient of morphed faces along the fear-happy dimension. We have acquired the following behavioral and neural data:3. Through a series of experiments4, we have characterized the LPP in detail and shown that it is specifically associated with behavioral judgments about choices that are ambiguous. This result is consistent with the literature showing that the LPP is sensitive to various types of ambiguity, including ambiguous facial expressions5, racially ambiguous faces6, socially relevant concepts7, affective pictures10, and stimulus uncertainty11.Electroencephalogram (EEG): Data from 97 unique neurotypical participants were acquired through a series of 8 experiments. An example result is the late positive potential (LPP), which encodes facial emotion ambiguity12, and activation of the right amygdala12, dorsomedial prefrontal cortex (dmPFC), and ventromedial prefrontal cortex (vmPFC)3 for categorical ambiguity of the stimuli. These results are consistent with the role of the amygdala in emotion processing15, especially fear processing18, as well as its role in detecting ambiguous stimuli and modulating vigilance and attention accordingly21. Furthermore, although the anterior cingulate cortex (ACC) has functional segregations (see23 for details), most of its functions involve processing ambiguity in some form, which requires conflict resolution, ongoing action monitoring, dynamic adjustments in cognitive control, and inversely correlates with confidence in judgment30 , consistent with our results.Functional magnetic resonance imaging (fMRI): Data from 19 neurotypical participants were acquired. Example results include activation of the left amygdala for the degree of fearfulness/happiness in the face33. By recording from 16 neurosurgical patients with depth electrodes , we have shown that neurons in the amygdala parametrically encode the degree of fearfulness/happiness in the face and the categorical ambiguity of the stimuli12, and neurons in the dmPFC encode the categorical ambiguity of the stimuli34. These results are consistent with the above fMRI results.Single-neuron recordings: We had the unique opportunity to record single-neuron activity directly from the human brain in neurosurgical patients undergoing monitoring to localize their epileptic seizures4 and face size35); and facial regions of interest (ROIs) have been delineated. Furthermore, we can compare eye movement in people with autism spectrum disorders (ASD) vs. controls (see below).Eye tracking: We acquired eye tracking data from 24 neurotypical participants. Fixations can be compared between task conditions .Autism spectrum disorder (ASD): We acquired behavioral and eye tracking data from 18 participants with ASD and 15 matched controls. We have shown reduced specificity in emotion judgment in participants with ASD12. This result is in line with other studies on amygdala lesions that have shown altered emotion recognition and judgment45.Amygdala lesion: We acquired behavioral data from 3 rare patients with focal bilateral amygdala lesions. We have shown an increase in behavioral sensitivity to fear in these patientsFaces are among the most ubiquitous social stimuli that people see in everyday life. Although there is a large literature studying face perception in humans, most of these studies focus on a single aspect of face perception using unimodal approaches. To comprehensively understand face perception, we highlighted the importance of multimodal cognitive neuroscience approaches for studying face perception46, computational modeling of single-neuron activity to explain autism behaviors47, and multimodal functional connectivity analysis34. Along with other multimodal face datasets (e.g.48), our current dataset will contribute to a more comprehensive understanding of face perception.With this dataset, further analysis can be performed to compare different participant groups  and different neural signals . Example usage of this dataset includes a confidence database37. Below, we provide an overview of our methods.The detailed methods have been described in our previous studies37, our data are considered adequate for detecting task effects and for making comparisons using different methodologies.In this study, we recruited adult participants of both sexes. There was no selection of participants based on age (adults only), sex, race, or ethnicity. The sample size for each experiment was determined empirically based on prior literature. Although we did not perform a formal power analysis to estimate the sample size, given our prior resultsIn the face judgment task with fear-happy morphed emotions, 23 neurotypical participants took part in the EEG experiment, 19 neurotypical participants took part in the fMRI experiment, 24 neurotypical participants took part in the eye tracking experiment, 16 neurosurgical patients (22 sessions) took part in in the single-neuron recording experiment, 18 high-functioning participants with ASD and 15 matched controls  took part in the eye tracking experiment, and 3 rare patients with focal bilateral amygdala lesions took part in the behavioral experiment 51 or Social Communication Questionnaire (SCQ)52 when an informant was available. The ADOS-2 was scored according to the latest algorithm, and we also derived severity scores for exploratory correlation analyses : 12.1 ± 4.22 (mean ± SD), restricted and repetitive behavior (RRB): 3.13 ± 1.36, severity score for social affect (CSS SA): 8.00 ± 1.71; severity score for restricted and repetitive behavior (CSS RRB): 7.13 ± 1.36, severity score for social affect plus restricted and repetitive behavior (CSS All): 7.88 ± 1.54). The ASD group had a full-scale IQ (FSIQ) of 105 ± 13.3 , a mean age of 30.8 ± 7.40 years, a mean Autism Spectrum Quotient (AQ) of 29.3 ± 8.28, a mean SRS-2 Adult Self Report (SRS-A-SR) of 84.6 ± 21.5, and a mean Benton score of 46.1 ± 3.89 . ADOS item scores were not available for two participants, so we were unable to utilize the revised scoring system. But these individuals’ original ADOS algorithm scores all met the cutoff scores for ASD. Fifteen neurologically and psychiatrically healthy participants with no family history of ASD  were recruited as controls. Controls had a comparable FSIQ of 107 ± 8.69  and a comparable mean age of 35.1 ± 11.4 years (P = 0.20), but a lower AQ  and SRS-A-SR  as expected. Controls were also matched on sex, race and education.Specifically, all 18 participants with ASD met DSM-V/ICD-10 diagnostic criteria for autism spectrum disorder, and met the cutoff scores for ASD on the Autism Diagnostic Observation Schedule-2 (ADOS-2) revised scoring system for Module 453. AM and BG are monozygotic twins, both with complete destruction of the basolateral amygdala and minor sparing of anterior amygdaloid and ventral cortical amygdaloid parts at a rostral level, as well as lateral and medial parts of the central amygdaloid nucleus and the amygdalohippocampal area at more caudal levels54. The details of the neuropsychological assessments of these patients have been described previously55. The anatomical scans of the lesions are shown in our previous report12.AP, AM and BG are three patients with selective bilateral amygdala lesions as a result of Urbach-Wiethe diseaseAll participants had normal or corrected-to-normal visual acuity. Participants provided written informed consent according to protocols approved by the Institutional Review Board (IRB) of the South China Normal University (SCNU), Huntington Memorial Hospital (HMH), Cedars-Sinai Medical Center (CSMC), and California Institute of Technology (CIT).56, a database of face images expressing highly recognizable emotions. The facial expressions were morphed by a computer algorithm; therefore, they are not real faces. For each individual, we selected unambiguous exemplars of fearful and happy expressions as evaluated with normative rating data provided by the database creators. To generate the morphed expression continua for this experiment, we interpolated pixel value and location between fearful exemplar faces and happy exemplar faces using a piece-wise cubic-spline transformation over a Delaunay tessellation of manually selected control points. We created five levels of fear-happy morphs, ranging from 30% fear/70% happy to 70% fear/30% happy in steps of 10%. Low-level image properties were equalized by the SHINE toolbox57  Fourier amplitude spectra, for normalizing and scaling mean luminance and contrast, and for exact histogram specification optimized for perceptual visual quality).Stimuli were morphed expression continua between exemplars of fearful and happy expressions . Similar to fear-happy morphs, we also selected 4 identities  and two Caucasian), with 2 unambiguous faces and 5 morph levels for each identity. The morphs ranged from 30% anger/70% disgust to 70% anger/30% disgust in steps of 10%.Anger-disgust morphs were created by FaceGen Modeller  of the face model. This task had the highest ambiguity because whether the face model is poor or rich could not be told without any priors.EEG Control Experiment 1 had a speeded response: participants were instructed to respond as quickly as possible. The stimulus stayed on the screen until the button press. Similarly, participants had 2 seconds to respond, otherwise, the trial was aborted and discarded. It is worth noting that in this task, the question prompt asking participants to make the best guess of the facial emotion preceded the stimulus and was presented for 500 ms. In EEG Control Experiment 1a,participants were asked to judge the EEG Control Experiment 2 modulated the ambiguity contexts: participants only judged anchor faces  in the first and third block  and judged both anchor and morphed faces in the second block .EEG Control Experiment 3 used anger-disgust morphed emotions. The task was the same as the fear-happy judgment task.EEG Control Experiment 4 modulated judgment responses/decisions. In EEG Control Experiment 4a (free viewing), participants were instructed to freely view the fear-happy morphed faces without pressing any buttons. In EEG Control Experiment 4b (unambiguous decisions), participants were instructed to discriminate whether the stimulus was a human or an animal, using ambiguous anger-disgust morphed faces and cat-dog morphed animals.Participants were seated comfortably about 1.1 m in front of a computer screen in a dimly lit and electromagnetically shielded room. Experiments were administered on a 19-in. (37.7 × 30.1 cm screen size) IBM LCD display (1280 × 1024 screen resolution). Stimuli were presented using E-prime. EEGs were recorded using a digital AC amplifier from 32 scalp sites with tin electrodes mounted in an elastic cap (NeuroScan4.5) according to the International 10–20 system. EEGs were recorded from the following sites: frontal: FP1, FP2, F7, F3, Fz, F4, F8; frontal-central: FC3, FCz, FC4; central: C3, Cz, C4; central-parietal: CP3, CPz, CP4; parietal: P7, P3, Pz, P4, P8; frontal-temporal-parietal: FT7, TP7, T7, T8, TP8, FT8; and occipital: O1, Oz, O2. The vertical-oculograms (VEOG) were recorded from left supra-orbital and infra-orbital electrodes. The horizontal electro-oculograms (HEOG) were measured from electrodes placed lateral to the outer canthi of the left and right eyes. The ground electrode was placed on the forehead. One reference electrode was placed at the left mastoid and the other at the right mastoid, and all recordings were referenced to the right mastoid. All impedance was maintained below 5ΚΩ. EEG and electro-oculogram (EOG) were amplified using a 0.05–70 Hz band-pass filter and were continuously sampled at 500 Hz for each channel. In Control Experiment 4, EEGs were recorded using a digital AC amplifier from 64 scalp sites.58 and in-house MATLAB functions. The continuous EEG data were re-referenced to the average of the left and right external mastoid signals to avoid biasing the data towards one hemisphere60. The data was filtered using a digital zero-phase shift band-pass filter of 0.5–30 Hz with a slope of 24 dB/octave. Then the continuous EEG data were epoched into 3-s segments  or 1-s segments . The pre-stimulus interval from –400 to –200 ms was used as the baseline for long epochs  and the pre-stimulus interval from –200 to 0 ms was used as the baseline for short epochs. The data were baseline corrected by subtracting the average activity during the baseline period.EEG data were processed using EEGLABhttp://erpinfo.org/erplab), in which vertical ocular artifacts exceeding a normalized cross-variance threshold of 0.7 were detected during the whole epoch61. We rejected these trials because blinks might not only alter the sensory input of that trial, but also contaminate the EEG signals, especially the signals from the frontal channels. Although we did not explicitly instruct participants not to explore the face during face presentation, the fixation cross preceding the stimulus could reduce eye movements and artifacts related to eye movements. Epochs with saccadic eye movements were detected and discarded using a step-like artifact-detection function, in which horizontal ocular artifacts exceeding 70 μV in amplitude were detected during the entire epoch with 200 ms moving window and 50 ms increment steps. This function is suitable to detect saccadic eye movements that typically consist of sudden, step-like changes in voltage61. Remaining artifacts were further detected using a moving-window peak-to-peak artifact-detection method on specific midline electrodes. Epochs were excluded if the peak-to-peak voltage  exceeded a threshold of 100 μV. Bad channels were interpolated using the average voltage from their surrounding electrodes. Notably, we repeated our artifact rejection using independent component analysis (ICA) and we derived qualitatively the same results.Trials that had blinks in any part of the segments were excluded using a blink detection tool from ERPlab  based on the time window of 400 to 700 ms after stimulus onset at the parietal-central (Pz) electrode63. Stimuli were presented using a mirror attached to the MRI head coil. Whole-brain data were acquired with echo planar T2*-weighted imaging (EPI), sensitive to blood-oxygen-level dependent (BOLD) signal contrast . T1 weighted structural images were acquired at a resolution of 1 × 1 × 1 mm.MRI scanning was conducted at the SCNU on a 3-Tesla Tim Trio Magnetic Resonance Imaging scanner  using a standard 12-channel head-coil system. Stimuli were presented to the participants on a back-projection screen using MATLAB with the Psychophysics Toolboxwww.fil.ion.ucl.ac.uk/spm/). The first 4 volumes were discarded to allow the MR signal to reach steady-state equilibrium. EPI images were sinc interpolated in time to correct for slice-timing differences and realigned to the first scan by rigid-body transformations to correct for head movements. Utilizing linear and nonlinear transformations and smoothing with a Gaussian kernel of full-width-half maximum 6 mm, EPI and structural images were coregistered to the T1 MNI 152 template . Global changes were removed by high-pass temporal filtering with a cutoff of 128 s to remove low-frequency drifts in signal.Neuroimaging data were preprocessed and analyzed using SPM12  design matrix, for every participant we estimated a GLM with autoregressive order 1 [AR(1)] and the following regressors (R): R1 at face presentation; R2 at face presentation modulated by fear levels: 100%, 70%, 60%, 50%, 40%, 30%, 0%; R3 at face presentation modulated by ambiguity levels: unambiguous, 30%/70% morph (intermediate ambiguity), 40–60% morph (high ambiguity); and R4 at fixation presentation. For all GLM analyses, six head-motion regressors based on the SPM’s realignment estimation routine were added to the model . Multiple linear regressions were then run to generate parameter estimates for each regressor for every voxel. The contrast  images of the first-level analysis were entered into one-sample 65. Bipolar wide-band recordings (0.1–9 kHz), using one of the eight microwires as reference, were sampled at 32 kHz and stored continuously for off-line analysis with a Neuralynx system . The raw signal was filtered with a zero-phase lag 300-3 kHz bandpass filter and spikes were sorted using a semiautomatic template matching algorithm66. Units were carefully isolated and spike sorting quality were assessed quantitatively.We recorded bilaterally from implanted depth electrodes in the amygdala and dmPFC  from patients with pharmacologically intractable epilepsy. Target locations were verified using post implantation structural MRIs. At each site, we recorded from eight 40 μm microwires inserted into a clinical electrode as described previouslyTo analyze the response of individual neurons, we only considered single units with an average firing rate of at least 0.2 Hz (entire task). Trials were aligned to face onset, and the baseline firing rate was calculated in a 1-s interval of blank screen right before face onset. To select neurons that showed a significant trial-by-trial correlation with the level of emotion ambiguity, we quantified the response of each neuron based on the number of spikes in a 1.5-s window starting 250 ms after stimulus onset. Peristimulus time histograms (PSTH) were computed by counting spikes across trials in consecutive 250 ms bins. Comparisons between morph/ambiguity levels in the PSTH were made using a one-way ANOVA at P < 0.05 and Bonferroni-corrected for multiple comparisons across bins.2. Fixations were defined as the complement of a saccade, i.e., periods without saccades, and the fixation locations were determined using the EyeLink event parser. Ten healthy participants were recorded using a remote noninvasive infrared Tobii T120 system which recorded binocular gaze at 120 Hz. The stimuli were presented using Matlab with the Psychophysics Toolbox. The Tobii visualization software (Tobii Studio 2.2) was used to record eye movements and perform gaze analysis. Fixations were detected by Tobii Fixation Filter implemented in Tobii Studio. The Tobii Fixation Filter is a classification algorithm proposed by Olsson67 and detects quick changes in the gaze point using a sliding window averaging method. The velocity threshold was set to 35 pixels/sample, and the distance threshold was set to 35 pixels in our study.In the eye tracking experiment with neurotypical participants conducted at the SCNU Table , we usedIn the eye tracking experiment conducted at the CIT focusing on studying ASD Table , 18 partTo quantitatively compare the fixation properties within certain parts of the face, we defined three regions of interest (ROIs): eyes, mouth, and center. Each ROI is a rectangle, and the eye and mouth ROI have the same size. To compute fixation density maps, fixation locations were smoothed with a 40-pixel 2D Gaussian kernel with a standard deviation of 10 pixels. The fixation density map indicates the probability of fixating a given location (in arbitrary units), which was calculated based on the number and duration of fixations and ensured an equal contribution from each participant and statistical independence between participants. The average fixation density within the ROIs was calculated for each participant and for each morph level during the entire stimulus period. Statistical comparisons were then performed to compare whether the mean fixation density, the total fixation duration, the mean fixation duration, the percentage of the number of fixations, and the latency of the first fixation falling into an ROI differed between fear/ambiguity levels, for each ROI.68 were sorted out in separate directories based on the data formats and experiments. A detailed description of the variables and usage of data and code can be found in the corresponding ‘README.docx’ file in each data directory.The data and codeThe stimuli used in this study are stored in the ‘/01 Stimuli/’ directory. Additionally, an independent group of 10 participants from the SCNU provided valence and arousal ratings for the fear-happy stimuli. Each face was rated five times on a scale from 1 to 10. For valence, participants were asked to rate how pleasant the displayed emotion of the face was, with 1 representing ‘very unpleasant’ and 10 representing ‘very pleasant’. For arousal/intensity, participants were asked to rate how intense the displayed emotion of the face was, with 1 representing ‘very mild/calm’ and 10 representing ‘very intense/excited’. The original behavioral data for each participant are stored in the corresponding ‘.DAT’ files, where ‘v_’ represents valence ratings and ‘a_’ represents arousal ratings. Additionally, a group summary of the ratings is stored in the file ‘FaceRatingSummary.xlsx’.Behavioral data from all participants  are stored in the file ‘behaviorData.mat’ in the ‘/02 Behavioral Data and Code/’ directory. Each element of the structure array corresponds to a specific participant. The code for the seven levels of fearfulness (ranging from 1 for the most fearful to 7 for the happiest) is stored in the field ‘codeL’. The code for the face identity is stored in the field ‘codeI’. The button press responses, where 1 represents a fearful response and 2 represents a happy response, are stored in the field ‘vResp’. Response times are recorded and stored in the field ‘RT’. Confidence ratings are stored in the field ‘vCR’. Response times for confidence ratings are stored in the field ‘RT_CR’. It is worth noting that EEG and fMRI participants as well as a subset of ASD, ASD control, and single-neuron participants did not provide confidence ratings.62 because the scalp topography of the difference waveform between high ambiguity and unambiguous stimuli showed the most pronounced difference at Pz in this time window3. The LPP waveforms for three levels of ambiguity can be obtained by running the script ‘LPP_plots_3levels_ambiguity_longepoch.m’ for the long-epoch data or ‘LPP_plots_3levels_ambiguity_shortepoch.m’ for the short-epoch data. The mean and peak amplitudes of the LPP are stored in the files ‘LPP_long.mat’ and ‘LPP_short.mat’, respectively. The mean and peak amplitudes for three levels of ambiguity can be plotted using the scripts ‘LPP_bars_longepoch.m’ and ‘LPP_bars_shortepoch.m’, which load the files ‘LPP_long.mat’ and ‘LPP_short.mat’, respectively. In addition, similar analyses can be performed for the N170 component using the script ‘N170_plots_3levels_ambiguity_shortepoch.m’.The preprocessed EEG data and code are stored in the ‘/03 EEG Data and Code/’ directory. The EEG data after artifact rejection using long epochs (–1000 to 2000 ms relative to stimulus onset) are stored in the file ‘Data_FearHappy_LongEpoch.zip’. The long epochs are better suited for time-frequency analysis. Additionally, the EEG data after artifact rejection using short epochs (–200 to 800 ms relative to stimulus onset) are stored in the file ‘Data_FearHappy_ShortEpoch.zip’. The short epochs are better suited for ERP analysis. Here, we focused on the LPP based on the time window of 400 to 700 ms after stimulus onset at the parietal-central (Pz) electrodeThe data and code for the control experiments are stored in a similar format in the same directory. Each control experiment is stored in a separate zip file.The preprocessed fMRI data are stored in the ‘/04 fMRI Data and Code/’ directory in two files: ‘sub1-10.zip’ and ‘sub11-19.zip’. The file ‘onsets.zip’ contains all the behavioral data and code necessary for extracting the stimulus onsets for the parametric design. The script ‘step1_firstlevel_model_face_fear_ambiguity3.m’ generates the first-level parametric model for each participant. The script ‘step2_firstlevel_contrasts_face_para3.m’ generates the first-level contrasts for each participant. The script ‘step3_secondlevel_model_face_para3.m’ generates the second-level (group average) contrasts. The file ‘rfx_face_emotion_para3.zip’ contains all the group-averaged contrasts. These steps are performed using SPM12 in MATLAB. The file ‘Brain Activation.docx’ documents all the coordinates of the activated brain regions.Single-neuron data from the amygdala and MFC are stored in the file ‘FiringRate_Amygdala.mat’ and ‘FiringRate_MFC.mat’, respectively, in the ‘/05 Single-neuron Recordings Data and Code/’ directory. Firing rates estimated within different time windows for each neuron are stored in the variable ‘FR’. For each neuron, the timestamps (in μs), session ID, channel ID, cluster ID, and recording brain area are stored in the variables ‘timestampsOfCellAll’, ‘vCell’, ‘vCh’, ‘vClusterID’, and ‘areaCell’, respectively . “sessions” contains the session identifiers corresponding to the index in ‘vCell’. The variable ‘beh’ stores the behavior for each recording session , and the variable ‘sessions’ stores detailed participant information. The file ‘plotRastersEachCell.m’ provides a demonstration of a raster plot for a given neuron. The file ‘plotGroupPSTH.m’ provides a demonstration of the group PSTH and a dot plot for group average.Eye tracking data are stored in the file ‘/06 Eye-tracking Data and Code/ET_Data_All.mat’. The eye tracking data for each trial were aligned to the image coordinates and then were extracted and stored in a matrix format in the variable ‘EM’. In each matrix, each row refers to a single fixation and the following saccade and the columns  refer to the following attributes: 1—trial index, 2—fixation index, 3—fixation coordinate in horizontal direction , 4—fixation coordinate in vertical direction, 5—fixation start time , 6—fixation end time, 7—type of stimuli (here we only included type ‘1’ for fear-happy morphs), 8—ROI that the fixation falls in, 9—the serial order of fixation in each ROI, 10—empty (not used), 11—empty (not used), 12—fixation duration, 13—saccade index, 14—saccade starting position in horizontal direction, 15—saccade starting position in vertical direction, 16—saccade ending position in horizontal direction, 17—saccade ending position in vertical direction, 18—saccade starting time, 19—saccade ending time, 20—saccade duration, 21—saccade distance , 22—saccade peak velocity , 23—saccade starting ROI , 24—saccade ending ROI, 25—serial order of saccade with starting position in this ROI , 26—serial order of saccade with ending position in this ROI, and 27—saccade direction . Note that data recorded using the Tobii eye tracker do not have the saccade information available.Furthermore, the variable ‘vSub’ denotes the participant category, the variable ‘beh’ stores the behavior , and the variable ‘sessions’ stores detailed participant information. It is important to note that all of these variables correspond to ‘EM’. Additionally, the variables ‘ROI_E’, ‘ROI_M’, and ‘ROI_C’ denote the ROI coordinates for the eyes, mouth, and center, respectively. The file ‘analyzeFixationDensityDemo’ plots fixation density maps for each participant group and the file ‘analyzeFixationAttributesDemo’ plots the percentage of the number of fixations, total fixation duration, mean fixation duration, and first fixation latency for each ROI. The file ‘analyzeFixationAttributesDemo’ also demonstrates comparisons between participant groups, across morph/stimulus levels, and across ambiguity levels.We first assessed the quality of our behavioral data and demonstrated that smooth psychometric curves could be derived from participants’ choices. Additionally, we showed that reaction time (RT) and participants’ confidence in their decisions varied as expected based on the stimulus.36 and amygdala lesion patients had shifted psychometric curves12).For each participant, we quantified behavior as the proportion of trials identified as fearful as a function of morph level Fig. . We founp2 = 0.45; EEG [Fig. p2 = 0.19; fMRI [Fig. p2 = 0.33). When grouping all trials into three levels of ambiguity, each group of participants showed the shortest RT for anchor faces (no ambiguity) and the longest RT for high ambiguity faces .We found that RT was faster for anchor faces  compared to ambiguous faces for each participant group MRI Fig. : Fp2 = 0.65; Fig. p2 = 0.76; Fig. p2 = 0.086; Fig. p2 = 0.139; Fig. Eye tracking participants reported their confidence in their decisions after reporting a face as fearful or happy. They reported significantly higher levels of confidence for anchor faces compared to ambiguous faces  = 42.0, P < 0.001, η−4, ηp2 = 0.34), and post-hoc t-tests revealed a significant difference between anchor  and intermediate ambiguity  = 3.17, P = 0.004, Cohen’s d = 0.66), and a marginal difference between intermediate and high ambiguity  = 1.73, P = 0.098, d = 0.36). In addition to the LPP, we also observed an N170 component in electrodes TP7 and TP8 in response to emotional facial images. However, the N170 was not sensitive to varying degrees of emotion ambiguity  = 0.406, P = 0.669, ηp2 = 0.018) or valence  = 0.659, P = 0.522, ηp2 = 0.029).In the fear-happy judgment task Fig. , we foun−10, ηp2 = 0.76).In Control Experiment 1a , the coding of stimulus ambiguity was relatively weaker compared with the emotion judgment task.In Control Experiment 1b .In Control Experiment 1c . Therefore, the LPP was influenced not only by the levels of ambiguity but also by the context of ambiguous stimuli.In Control Experiment 2 .In Control Experiment 3 .In Control Experiment 4a .In Control Experiment 4b  a significant increase of BOLD signal in the bilateral inferior frontal gyrus (IFG)/anterior insula and dorsal medial prefrontal cortex (dmPFC)/dorsal anterior cingulate cortex (dACC) as a function of increasing emotion ambiguity Fig. , (2) a s−6; see Fig. −6; see Fig. We recorded from 321 neurons in the amygdala (21 sessions) and 236 neurons in the dmPFC . To validate our data, we demonstrated that the responses of amygdala and dmPFC neurons were modulated by the level of emotion ambiguity. Specifically, we used a linear regression to identify neurons whose firing rate correlated trial-by-trial with three levels of emotion ambiguity. We found 36 amygdala neurons , the same figures stored in the ‘output figures’ folder can be obtained.


To examine the causal associations of diet-derived circulating antioxidants with knee osteoarthritis (OA), hip OA, and rheumatoid arthritis (RA) within the two-sample Mendelian randomization (MR) framework.Independent single-nucleotide polymorphisms (SNPs) significantly associated with circulating levels of diet-derived antioxidants  were extracted as genetic instruments. Summary statistics of genetic instruments associated with knee OA, hip OA, and RA were obtained from corresponding genome-wide association studies (GWASs). The inverse-variance weighted (IVW) was applied as the primary analysis method, with four sensitivity analysis approaches employed to evaluate the robustness of the primary results.p = 4.43 × 10−3], while genetically determined per unit increase in absolute circulating levels of β-carotene was suggestively associated with increased risk of RA . No other causal association was found. Significant evidence for heterogeneity and pleiotropic outlier was only identified when absolute circulating vitamin C was evaluated as the exposure, whereas all sensitive analysis provided consistently non-significant results.Genetically determined per unit increment of absolute circulating levels of retinol was significantly associated with a reduced risk of hip OA [odds ratio (OR) = 0.45, 95% confidence interval (CI) 0.26–0.78, Our results demonstrated that genetically determined lifelong higher exposure to absolute circulating levels of retinol is associated with a decreased risk of hip OA. Further MR study with more genetic instruments for absolute circulating levels of antioxidants are needed to confirm our results. As the two most common forms of arthritis, osteoarthritis (OA) and rheumatoid arthritis (RA) affect over 300 million and nearly 20 million people worldwide, respectively. It is foreseeable that these figures will dramatically increase as a result of the prolonged life expectancy as well as the ageing of the global population . OA maniOf note, a growing body of evidence indicates that oxidative stress associated with the abundant production of reactive oxygen species (ROS) is involved in the onset and progression of OA and RA, suggesting that diet-derived antioxidants might possess the preventive potential against OA  and RA . OxidatiRecently, Mendelian randomization (MR), employing genetic variants such as single nucleotide polymorphisms (SNPs) as instrumental variables (IVs) in observational settings to make causal inference between environmentally modifiable exposure and outcome, has been gaining popularity in recent years . Since gThe present two-sample MR study was conducted based on publicly available genome-wide association studies (GWASs) summary-level data, and ethical approval and informed consent had been acquired in each original study. In brief, we first extracted the summary statistics of genetic instruments for absolute circulating antioxidants and circulating antioxidant metabolites from the corresponding GWASs, then the summary statistics for these genetic instruments were obtained from GWASs of knee OA, hip OA and RA. Eventually, the causal associations of diet-derived circulating antioxidants and risk of knee OA, hip OA, and RA were evaluated within a two-sample MR framework, and the schematic overview of our present study design is shown in r2 < 0.001 within a 10,000 kb sliding window) serving as IVs significantly associated with diet-derived circulating antioxidants were extracted.In our present two-sample MR study, a total of five common diet-derived antioxidants, namely retinol, β-carotene, lycopene, vitamin C and Vitamin E (α-tocopherol and γ-tocopherol), were included. For each diet-derived antioxidant, the absolute circulating level and/or relative circulating level of the antioxidant metabolite were considered. Summary-level genetic association statistics for independent SNPs (p < 5 × 10−8) in the meta-analysis of a GWAS and further replication studies comprising 8,902 Caucasian individuals were selected (p < 5 × 10−8) in meta-analysis of GWAS and further replication samples involving 3,881 European individuals was selected (p < 5 × 10−6) were obtained from a GWAS of 441 Old Order Amish adults (p < 5 × 10−8) in a large-scale GWAS in 52,018 individuals of European ancestry were chosen (p < 5 × 10−8) in meta-analysis of one GWAS with two independent replication samples consisting of 7,781 European individuals were obtained , α-tocopherol (n = 11), and γ-tocopherol (n = 13) at suggestive genome-wide significance level (p < 1 × 10−5) were derived from a GWAS comprising 7,824 adults from 2 European population studies (p < 1 × 10−5) were derived from a recently published GWAS comprising 1,957 European participants  and hip OA  were extracted from one recent GWAS meta-analysis performed based on the United Kingdom Biobank and Arthritis Research UK Osteoarthritis Genetics (arcOGEN) resources, in which approximately 17.5 million SNPs in 455,221 individuals of European ancestry were analyzed  based on a type I error being 0.05, a statistical power of 0.8, sample size, proportion of case and variance explained by genetic instruments (2R) were estimated using an online toolR2 were either extracted from the original studies or calculated based on the following formula: R2 = 2 × EAF × (1 − EAF) × beta2/ [(2 × EAF × (1 − EAF) × beta2 + 2 × EAF × (1 − EAF) × N × se2)], where EAF, beta, se, and N denote the effect allele frequency, estimated effect and its standard error of genetic instrument on exposure, and sample size, respectively , set based on Bonferroni correction method, was considered statistically significant, while a two-sided p value less than 0.05 was regarded suggestive significance. All analysis were performed utilizing “forestplot,” “TwoSampleMR” and “MRPRESSO” packages in R version 4.2.0.The inverse-variance weighted (IVW) was employed as the primary MR analysis approach, and four sensitive analysis methods, including weighed median, weighted mode, MR-Egger and MR pleiotropy residual sum and outlier (MR-PRESSO), were applied to test the robustness of IVW estimates to horizontal pleiotropy. When all selected SNPs are valid instrumental variables, the IVW method gives the most precise estimate of the causal effect by combing the Wald ratio estimate of each genetic instrument in a meta-analysis model. When only one genetic instrument is available, the Wald ratio estimate was applied. The weighted median method could provide a consistent causal estimate if at least one half of the employed genetic instruments are valid , while tectively . The eff2R) for absolute circulating levels of retinol, β-carotene, lycopene, vitamin C, and α-tocopherol was 2.3, 2.5, 31.0, 1.7, and 1.7%, respectively. The R2 for circulating metabolites levels of retinol, vitamin C, α-tocopherol, and γ-tocopherol was 22.9, 21.7, 6.8, and 9.8%, respectively. The effect estimates for the associations of genetic instruments for absolute circulating antioxidants and circulating antioxidant metabolites with knee OA, hip OA and RA are shown in p = 4.43 × 10−3), while genetically determined per unit increase in β-carotene was associated with increased risk of RA  with suggestive significance. Since the number of genetic instruments available for retinol and β-carotene was less than 3, sensitive analysis could not be performed. As for the associations of the other three diet-derived antioxidants  with knee OA, hip OA and RA, no significant evidence was found in the main IVW analysis, and the estimates derived from all sensitive analysis methods were consistently non-significant. Significant heterogeneity across individual genetic instrument estimate was found for the association of vitamin C with knee OA (p = 2.09 × 10−3), hip OA (p = 0.03) and RA (p = 1.91 × 10−3). No significant evidence of pleiotropy was found in MR-Egger regression  study (n = 108)  . As regan = 108) , 32, whin = 108) , 21, 22.n = 108) , while an = 108) . Besidesn = 108)  as well n = 108) . It has n = 108) . Of inten = 108)  found thn = 108) , it is on = 108)  also focn = 108) . These mn = 108) , and ourThe main strength of our present study lies in the utilization of genetic instruments as proxies for diet-derived circulating antioxidants evaluated as absolute levels and relative concentrations in the form of metabolites to perform two-sample MR analysis to infer their causal effects on risk of knee OA, hip OA and RA. However, several potential limitations should be acknowledged. First, since only less than three genetic instruments are available for absolute circulating levels of retinol and β-carotene, we could not perform sensitive analysis to examine and correct for the potential pleiotropy. Nevertheless, these genetic instruments are not associated with other factors as shown in PhenoScanner database, suggesting that the possibility of pleiotropy is small. Second, non-linear causal associations of diet-derived circulating antioxidants with OA and RA could not be examined based on the summary-level data. However, it is reasonable to speculate that insufficient or excessive levels of antioxidants might have a harmful health effect. Third, our present MR analysis was underpowered to detect small causal effects due to the small proportion of variance explained by currently available genetic instruments for absolute circulating levels of all included antioxidants except for lycopene. Notwithstanding, consistently non-significant evidence was shown for circulating metabolites with sufficient statistical power. Last, although most of the causal associations detected in our present MR analysis were non-significant, we could not rule out the possibility that diet-derived antioxidants might have beneficial health effects in individuals deficient in these antioxidants. Besides, a synergistic effect between diet-derived antioxidants might exist, whereas this possibility could not be explored in our present MR study.Taken together, utilizing genetic instruments as proxies for diet-derived circulating antioxidants evaluated as absolute levels and relative levels of their metabolites, the causal associations of diet-derived circulating antioxidants with the risk of knee OA, hip OA and RA were comprehensively assessed within a two-sample MR framework. We found that genetically predicted higher absolute circulating levels of retinol are associated with a reduced risk of hip OA, while genetically predicted higher absolute circulating levels of β-carotene are associated with increased risk of RA. Further MR study with more genetic instruments for absolute circulating levels of antioxidants, followed by the findings in larger-scale GWASs in the future, are needed to confirm our results.The original contributions presented in the study are included in the article/Supplementary Material, and further inquiries can be directed to the corresponding author.Ethical approval was not provided for this study on human participants because the present two-sample MR study was conducted based on publicly available genome-wide association studies (GWASs) summary-level data, and ethical approval and informed consent had been acquired in each original study.YM, JL, and HC had full access to all of the data in the study and took responsibility for the integrity of the data and the accuracy of the data analysis. LH, YM, JL, and HC contributed to conception and design. LH, YX, TJ, ZZ, MW, and HC were involved in the acquisition of data. LH, YM, JL, and HC carried out analysis and interpretation of data. All authors were involved in drafting the article or revising it critically for important intellectual content, and approved the final version to be submitted for publication.This work was supported by Zhejiang Provincial Natural Science Foundation of China (Grant No. LY20H260001) and K.C. Wong Magna Fund in Ningbo University.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Decreased participation and complex transitions into adulthood among youth with disabilities may impede their well-being. To advance knowledge on the co-occurrence of mental health problems and physical disability, this brief report describes the frequency of mental health problems, measured by the Behavior Assessment System of Children (BASC-3), among transition-aged youth (14–25 years) with physical disabilities and examines the association between mental health problems and sex, age, and number of functional issues.Thirty-three participants completed a demographic questionnaire and the BASC-3. Frequency of BASC-3 scales falling within 3 categories: “within norms”, “at risk”, and “clinically significant” were described. Crosstabs and Chi-square tests were used to examine the association between BASC-3 scales and sex, age (< and ≥ 20), and number of functional issues (< and ≥ 6).Overall, “somatization”, “self-esteem”, “depression” and “sense of inadequacy” were the most common subscales being at risk. Participants with a higher number of functional issues (≥6) were more likely to fall within “at risk” or “clinically significant” categories across 20 (out of 22) BASC-3 scales, and female participants tended to fall more within “at risk” or “clinically significant” categories for 8 of BASC-3 scales. Younger participants (<20) were ranked in the “at risk” or “clinically significant” categories for 7 scales.Findings lend further support for the occurrence of mental health problems emerging in youth with physical disabilities and highlight initial trends especially across functional levels. Further investigation of such co-occurrences and the factors that affect their development is needed. Children and youth with physical disabilities often experience lower levels of participation compared to their peers without physical disabilities –4. DecreAlong with decreased participation, youth with physical disabilities also experience more complex transitions into adulthood that may adversely affect their quality of life and well-being. Adolescence and young adulthood are marked by pronounced psychological and physical changes . Mental Evidence on the co-occurrence of physical disabilities and mental health problems is emerging. A meta-analysis of 8 studies among children and youth with cerebral palsy conducted by Downs et al.  suggestsTo better understand the occurrence of mental health problems among transition-aged youth living with physical disabilities, this brief research report aimed to describe the frequency of mental health problems, as measured by the BASC-3, among youth aged 14–25 years with physical disabilities who experience different levels of mobility restriction in a large Canadian province where both English and French are common spoken languages. It also aimed to describe the frequency of mental health problems according to sex, age, and complexity of youth's condition in terms of number functional issues among this population.This brief research report presents baseline results  of a clinical trial that examined the effect of an intervention aimed to improve participation in leisure community-based activities on outcomes at the body function level using the specific subscales of the BASC-3 among other motor-related outcomes. As such, the present study is cross-sectional in nature. A total of 33 youth aged 14 to 25 years old (mean age: 20.24) were included from both sexes who spoke either English or French. Youth with a physical disability, presenting with restricted mobility , as well as multiple diagnostic categories were included. Participants were excluded if they were in their first year of recovery from severe brain injury or orthopedic surgery or if they were recovering from botulinum toxin treatment 6 months prior to the study or anticipated during the study. Youth with neurodegenerative disorders and/or significant intellectual delays were also excluded, as the latter had the potential to impact self-completion of any necessary forms and assessments.via Research Electronic Data Capture (RedCap) from all participants followed by the completion of a demographic questionnaire. The full BASC-3 was then completed online by the youth through the Q global platform . The BASC-3 was used to measure behaviors and emotions that pertain to aspects of mental health. It is a multicomponent instrument that takes self-reported measures to analyze behavioral and emotional aspects of children and youth (2–25 years old) . Two difDemographic questionnaire. The demographic questionnaire contained 14 different questions created by the research team. Listed questions were related to sex, age, type of community, spoken languages, living situation, education, engaged activities, functional issues, health conditions, rehabilitation services received  and mental health services received . More specifically, the current study looked at age, sex, and functional issues within the questionnaire. Age was noted in years and sex was categorized as male or female. To capture the complexity of participants' condition, as used in a study by Anaby et al.  to predip-value of Pearson Chi-Square, but when Chi-Square's assumption of cell counts with less that 5 or less than 20% was violated, the p-value of Fisher's Exact Test was used (p < 0.05.Descriptive statistics were used to report the frequency (presented in percentages) of observed mental health problems falling in categories “within norms”, “at risk”, and “clinically significant” for the entire sample. A bar chart was created to illustrate the percentage of participants who ranked “at risk” and “clinically significant” for each BASC-3 scale. A fourth category named “at risk and above” was created by combining “at risk” and “clinically significant” categories. To examine the association between the presence of mental health problems and variables of sex, age and number of functional issues, crosstabs and Chi-Square or Fisher Exact tests were performed. To do so, the data was divided into 2 subgroups for variables of sex , number of functional issues (<6 and ≥6) and age <20 and ≥20) based on the median. Chi-Square tests were performed to generate 2 × 2 crosstabulation tables to explore the relationship between 2 BASC categories (“within norms” and “at risk and above”) of each BASC-3 subscale and the 2 categories of each of the 3 variables . The significance of association between subscales and the 3 variables were determined using the  and ≥20 was used . Trends Thirty-three youth aged 14–25 years  participated in the study. Nineteen youth reported types of services received; 5 (26.4%) of which reported not receiving neither physiotherapy, occupational therapy, counseling, nor speech-language pathology and 14 (73.6%) participants responded to receiving at least one of these services. The number of services received ranged from 0 to 4 (median and mode were 1), with occupational therapy being the most common service received. Eight reported receiving counseling. The majority (90%) lived with at least one of their family members and the remaining (10%) lived alone or with their significant other. Youth reported up to 11 functional issues . The two most frequently reported functional issues were moving around (69.7%) and managing emotions (68.8%). Youth reported 1–6 health conditions . The two most frequent health conditions were orthopedic/movement difficulties (69.7%) and health impairments (24.2%) as detailed in n = 33):Frequency of occurrences of self-reported mental health problems across BASC-3 categories  closely followed by sensation seeking (84.8%), while the least common ones were somatization (48.5%) and sense of inadequacy (57.6%). With respect to the composite scales, inattention/hyperactivity (84.8%) followed by internalizing problems (63.6%) ranked most frequently “within norms” while the least common ones were emotional symptoms index (54.5%) and personal adjustments (57.6%). With respect to content scales, mania (75.8%) ranked most frequently “within norms” followed by anger control (72.7%) while ego strength (51.5%) and test anxiety (66.7%) were the least common ones.For the BASC-3 subscales, as shown in The most frequent “clinically significant” BASC-3 subscales were somatization (30.3%) followed by depression (21.2%) whereas the least common ones were attention problems (3.0%) and hyperactivity (3.0%). With respect to the composite scales, internalizing problems (18.2%) and emotional symptoms index (18.2%) ranked most frequently “clinically significant” while the least common one was personal adjustments (12.1%). None of the scores pertaining to inattention/hyperactivity fell within the “clinically significant” category. As for the content scales, anger control and ego strength (12.1%) ranked most frequently “clinically significant” while test anxiety (3.0%) was the least common one. None of the scores pertaining to mania fell within the “clinically significant” category.In total, participants ranked most frequently “at risk and above” for self-esteem (54.5%) closely followed by somatization (51.5%) while least frequently for hyperactivity (12.1%) and sensation seeking (15.2%). With regards to the composite scales, emotional symptoms index (45.5%) closely followed by personal adjustments (42.4%) ranked most frequently “at risk and above” while the least common ones were inattention/hyperactivity (15.2%) and internalizing problems (36.4%). With respect to content scales, ego strength (48.5%) and test anxiety (33.3%) ranked most frequently “at risk and above” while the least common ones were mania (24.2%) and anger control (27.3%).Among female participants, atypicality, sense of inadequacy, self-esteem, test anxiety, and ego strength were most frequently ranked “at risk” (45%), while somatization was most frequently ranked “clinically significant” (45%) and “at risk and above” (70%). Within the male participants, emotional symptoms index was the most frequent scale ranked “at risk” (30.8%), while self-esteem was the most frequent subscale ranked “clinically significant” (23.1%) and “at risk and above” (46.2%).Overall, nine scales indicated a trend with sex, among which three were statistically significant see . Female Among the sub-group of participants with higher number of functional issues (≥6), atypicality, self-esteem, test anxiety and ego strength were most frequently ranked “at risk” (41.2%), while somatization was most frequently ranked “clinically significant” (47.1%) and “at risk and above” (70.6%). Amongst participants with lower number of functional issues (<6), sense of inadequacy, self-esteem, emotional symptoms index, personal adjustments, and ego strength were most frequently ranked “at risk” (31.3%), while self-esteem was most frequently ranked “clinically significant” (18.8%) and “at risk and above” (50.1%).In general, the crosstab analysis revealed that descriptively those with greater number of functional issues fell within the “at risk and above” category for most of the BASC-3 scales (20/22). The association between number of functional issues and presence of mental health problems (ranked “at risk and above”) was statistically significant in six scales. Specifically, youth with higher number of functional issues tended to experience depression, somatization, inattention/hyperactivity, problems with interpersonal relations, and to internalize problems, whereas those with lower number of functional issues were significantly more likely to seek sensations.For younger participants, aged below 20 years, anxiety, personal adjustments, mania, and ego strength were most frequently ranked “at risk” (42.9%), while depression, somatization, self-esteem, and anger control were most frequently ranked “clinically significant” (21.4%), and somatization and self-esteem “at risk and above” (57.1%). Within participants aged 20 and above, self-esteem was most frequently ranked “at risk” (36.8%), while somatization was most frequently ranked “clinically significant” (36.8%) and self-esteem “at risk and above” (52.6%).Overall, a descriptive trend between the presence of mental health problems and age groups was observed in seven BASC-3 scales, where younger participants (aged below 20 years) were more likely to fall within the “at risk and above” category than the older group (aged 20 years and above). The BASC-3 subscales that showed significant association with age were hyperactivity and mania, revealing that younger participants were significantly more likely to fall within the “at risk and above” category than the older group see . A similThis brief research report provides additional evidence for the occurrences of emerging mental health problems, measured by a comprehensive self-reported assessment (BASC-3), among transition-aged youth with physical disabilities. It also describes these co-occurrences across sex, age, and functional issues. Overall, when looking at the entire sample, “somatization”, “self-esteem”, “depression” and “sense of inadequacy” were the most common areas where youth reported difficulties, falling within the “at risk” or “clinical significant” categories. Similarly, Gorter et al.  found thSeveral associations (or trends) were observed between mental health issues and personal factors with a more pronounced association with complexity of the youth's condition. Specifically, youth with higher number of functional issues were more likely to fall within the “at risk and above” category for most of the BASC-3 scales, except for hyperactivity and sensation seeking. Although these associations were not statistically significant across all scales, it is important to reflect on the identified trends as prior research has shown similar findings. For example, Lindén-Boström & Persson  found thSome initial trends were observed across the sexes where nine scales of the BASC-3 showed a trend or an association with sex. Specifically, female adolescents, in comparison to male participants, were significantly more likely to fall with the “at risk and above” category on eight scales including self-esteem, test anxiety, atypicality and somatization. Our findings are similar to what has been establish in literature, showing sex differences for mental health in general . FemalesAge also appeared to be an important factor in some of the BASC domains. Results demonstrated that participants in the younger age group ranked more “at risk and above” compared to participants in the older age group across 7 (out of 22) subscales. Similarly, a study by Brossard-Racine et al.  found thOur results, based on the BASC-3 clinical norms, suggest that youth with physical disabilities are at a greater risk of mental health problems, evident in 12% to 54% of our sample  and comparable to emerging findings , 17. TheGeneralizability of the findings towards the larger population of youth with physical disabilities may be impacted by sampling strategy (convenience sampling) as well as inclusion and exclusion criteria, where individuals presenting with an intellectual delay, or a neurodegenerative disorder were excluded. When looking at data analysis, given that sampling distribution has an approximate chi-square distribution, a small sample size can impact accuracy of the approximation . Further


In the published article, there was an error in Abstract, subsection “Methods.” This sentence previously stated:A correction has been made to the Section “Two thousand four hundred and seventy three participants completed six-follow-up surveys in 2014 and were analyzed in this study.”The corrected sentence appears below:“Two thousand six hundred and thirty participants completed six-follow-up surveys in 2014 and were analyzed in this study. ”Abstract, subsection “Results.” These sentences previously stated:A correction has been made to the Section “Of the 2,473 participants, 14.1% were consistent daily tea drinkers, and 22.6% reported frailty at the 6-year follow-up. Compared to non-tea drinkers, consistent daily tea drinkers reported a significantly lower ratio of having frailty , adjusting for sociodemographic characteristics, health behavior, socioeconomic status, and chronic illnesses. In further subgroup analyses, consistent daily tea consumption significantly reduced the risk of frailty for males  but not females ; in the young  but not in the oldest (aged ≥ 80) ; informal education  but not formal education ; financial dependence  but not financial independence .”The corrected sentences appear below:“Of the 2,630 participants, 15.3% were consistent daily tea drinkers, and 22.6% reported frailty at the 6-year follow-up. Compared to non-tea drinkers, consistent daily tea drinkers reported a significantly lower ratio of having frailty , adjusting for sociodemographic characteristics, health behavior, socioeconomic status, and chronic illnesses. In further subgroup analyses, consistent daily tea consumption significantly reduced the risk of frailty for males  but not females ; informal education  but not formal education ; financial dependence  but not financial independence . Tea consumption was associated with a lower risk of frailty in both the young  and the oldest (aged ≥ 80) .”Materials and methods, subsection “Study sample,” paragraph 2. This sentence previously stated:A correction has been made to the Section “Finally, 2,473 participants completed a six-follow-up survey in 2014 and were analyzed in this study.”The corrected sentence appears below:“Finally, 2,630 participants completed a six-follow-up survey in 2014 and were analyzed in this study.”Materials and methods, subsection “Definition of frailty,” paragraph 1. This sentence previously stated:A correction has been made to the Section “The scale of a frailty index in this study has a Cronbach's alpha of 0.867.”The corrected sentence appears below:“The scale of a frailty index in this study has a Cronbach's alpha of 0.870.”Materials and methods, subsection “Definition of frailty,” paragraph 1. This sentence previously stated:A correction has been made to the Section “The participants with a score of >0.21 were referred to as frail (32), while those with a score of <0.21 were referred to as not frail.”The corrected sentence appears below:“The participants with a score of >0.21 were referred to as frail (32), while those with a score of ≤0.21 were referred to as not frail.”Results, paragraphs 1, 2, 3 and 4. These sentences previously stated:Corrections have been made to the Section “Characteristics of the participants at baseline are summarized in “P = 0.156), occupation (P = 0.381), and exercise (P = 0.392).”““Compared to non-tea drinkers, consistent daily tea drinkers had a significantly lower ratio of frailty  in the crude and  adjusted models . HoweverThe corrected sentences appear below:“Characteristics of the participants at baseline are summarized in “P = 0.214), occupation (P = 0.232), and exercise (P = 0.565).”““Compared to non-tea drinkers, consistent daily tea drinkers had a significantly lower ratio of frailty  in the crude and  adjusted models . HoweverThe authors apologize for these errors and state that these do not change the scientific conclusions of the article in any way. The original article has been updated.


Coverage and capacity are optimized in fifth generation (5G) networks by small base station (SBS) distribution in the coverage realm of macro base station (MBS). However, system performance is significantly reduced by inter-cell interference (ICI) because of the orthogonal frequency division multiple access assumption. In addition to ICI, this work considers intentional jammers’ interference (IJI) due to the presence of jammers. These Jammers try to inject undesirable energies into the legitimate communication band, which significantly degrade uplink (UL) signal-to-interference ratio (SIR). To reduce ICI and IJI, in this work, we employ SBS muting, where the SBSs near MBS are switched off. To further mitigate ICI and IJI, we use one of the effective interference management schemes a.k.a reverse frequency allocation (RFA). We presume that due to mitigation in ICI and IJI, the UL coverage performance of the proposed network model can be further improved. Heterogeneous cellular networks (HetNets) is a promising candidate technology for the future fifth generation 5G) networks –3. The wG networkIntra-cell interference(ICI) is still the key limiting factor in HetNets despite the adoption of orthogonal frequency division multiple access (OFDMA), which results in minimal intra-cell interference , 11.The functioning of the HetNet network can be negatively impacted by severe intentional jammers’ interference (IJI) caused by jammers’ presence –14. The SBS muting is taken into consideration in HetNets because I a user receives more coverage close to the MBS  and (ii)We refer to SBS muting by non-uniform HetNets (NUHs) and without SBS muting by uniform HetNets (UHs) in the remaining sections of the work. Different interference mitigation strategies, including reverse frequency allocation (RFA) , cell raDifferent key 5G technologies with their applications are presented in , 23. LatThe works in , 28 evalTheir proposed model leads to significant performance superiority due to lower IJI and ICI. The above-mentioned work, however, lacks to investigate both SBS muting and RFA scheme.The work in  study thIn , the autThe latest work on RFA employment can be found in , 21, 36,In this work, we look into HetNets’ performance in terms of coverage when there is IJI and ICI. To alleviate the impact of ICI and IJI we use SBS muting as well as RFA as a preventive measure for interference mitigation.A1, and the edge region, designated A2, with radii Δs1 and Δs2, accordingly [The paper uses two layers of BS, known as MBSs and SBSs, to illustrate a model based on HetNets. IJI attacks frequently result in additional UL intersections in addition to the typical ICI. Thus, the system as a whole is considered degenerative. In ordingly , 38.The significance of this work from the state-of-the-art can be summarized as follow.The work in –29 evaluIn contrast to our work, –32 lacksThe latest work of –35 evaluThe latest work on RFA employment can be found in , 21, 36.The following are this paper’s significant contributions.A2 when IJI and ICI are present.Analysis of the UL coverage for the typical user which is defined as Slivnyak theorem states that the statistical characteristics of an independent homogeneous Poission Point Process (IHPPP) are preserved and simplified by a typical user at origin , 40., U,This study examines how proactive interference control strategies can reduce IJI and ICI. RFA and the use of NUHs, a smart network design technique.For (i) UHs with RFA employment (see ) and ii), we devThe outcomes are presented for various network characteristics, including SIR threshold, jammers’ density and transmitted power, users’ transmit power and density of SBS.The remainder of the paper is laid out as follows. The system model is presented in Section 2. The suggested model’s coverage probabilities are calculated in Section 3. Section 4 contains the results and commentary. The paper is finished in Section 5. This section presents the suggested network design as shown in ϕM, ϕS, ϕu, and ϕJ, respectively [ζM, ζS, ζu, and ζJ, respectively. The proposed network models are presented in A2. The path loss exponent is denoted by α [h|2 ∼ exp(1) [h|. For RFA and NUH employment, we divide the MBS coverage region in to A1 and A2 with radii Δs1 and Δs2, respectively [In this paper, we consider two-tier HetNet comprising of co-deployed SBS’s with MBS’s. We suppose that there exist intentional jammers throughout the network which degrade the desired communication link. MBS’s, SBS’s, users, and jammers are distributed via IHPPPs ectively . The dented by α , 42. The∼ exp(1) , 40 is rectively , 43.Jammers are considered to transmits unwanted energy across the entire spectrum of the communication system to reduce network performance , 38, 44.g ∈  and l ∈  used alternatively. Due to efficient interference mitigation, RFA-based resource partitioning significantly improves coverage . By usin1 and F2, such that F = ⋃z∈ Fz, as shown in 1 and F2 of MBS is used for UL and DL communication in outer area macro cell  uplink coverage probability for uniform HetNets (UHs’) is presented in Subsection 3.1 while (ii) the uplink coverage probability in case of non-uniform HetNets (NUHs’) is derived in Subsection 3.2.This section focus on the assessment of coverage probability in the proposed network scenarios where ν in A2 can be obtained as:The UL coverage probability when there are intentional jammers (IJs) and RFA, A2, i.e., A1, i.e., IJ,A. Therefore, Following the architecture of RFA, the total interference in UL is the addition of the UL interference from MBSs in ν UL transmission power connected with MBS, Pt,j is the emitting power of jammers. Moreover, substituting Pt,lUL iss, where a + b) = exp(a) × exp(b).Here, Step (1) follows from the coverage probability definition , 40. SteA2, i.e., The Laplace transform (LT) of interference in UL from MBSs in a) follows the definition of LT [b) is achieved by substituting a), Step (c) is achieved by replacing s, s.t., b), Step (e) is followed by evaluating the LT of Step (d) with respect to hj, Step (f), is followed by considering probability generating functional (PGFL) of IHPPP [g) is achieved by replacing f), and Step (h) is achieved from Gauss-hypergeometric approximation of Step (g) [Here, Step  .A1, i.e., Similarly, the LT of the total UL interference received from the MBSs in A1, i.e., η2 is the ratio of In addition, the LT of the DL interference from SBSs in y as far , and is ν in A2 can be written as [The UL coverage probability, itten as PA2UL,*, an, an7), aessed as .A1 is muted and user in that vicinity is in coverage with MBS. The UL coverage probability, ν in A2 can be written asNon-uniform heterogeneous network deployment is established where SBS in By substituting  and 8)  8) into essed as . In 10)PA1UL(ΓM)essed as , J(·) inA = π(500m)2, s.t., A = A1UA2. Transmitted power by MBS, SBS, jammers, and users are supposed to be 40 dBm, 30 dBm, 20 dBm, and 20 dBm, respectively. Various network parameters such as ζM, ζS, ζj, ΓM, Pt,J and A2.This section describes results for the user’s UL coverage probability while taking into consideration: (i) UL coverage probability of UH and (ii) UL coverage probability of NUH. MATLAB 2015a has been used in drawing our results. MBS, SBS, jammers and users are dispersed in M in A2. This figure assumes ζj = 0 and 100, for both UH and NUH network scenarios. This figure indicates that the simulation results will coincide with the numerical results both for UH and NUH. The plots in the figure further demonstrate that NUHs with ζj = 0 lead to the highest coverage gain as compared with the rest of the scenarios. This is due to improved interference mitigation by NUHs as a result of lower SBS deployment.M for both UH and NUH in A2. This figure is obtained for ζj = 0 and 100 and ζS/ζM = 30. This result demonstrates that NUH with RFA outperforms the other scenarios due to significant interference mitigation. At ΓM = −10dB, the proposed NUH with RFA and ζj = 0 leads to 20% UL coverage gain.In M for UH and NUH, respectively. The plots in both the figures are obtained for ζj = 0, 100, 200, 300, 400, 500. Moreover, the results indicate that a sufficient number of IJs in the network are needed to significantly degrade UL coverage because of the wideband nature and low transmission power of IJs. Furthermore, increasing the value ζj leads to lower UL coverage in both UH and NUH due to higher interference. The results indicate significant coverage performance improvements by RFA and NUH due to effective interference mitigation.In ζj, while considering RFA, UH, and NUH. The plots are obtained for ΓM = 0 dB, −5 dB, −10 dB, −15 dB, −20 dB, −25 dB and ζS/ζM = 30. The plots indicate that higher values of ΓM lead to lower coverage due to lower user association. Furthermore, the plots in both figures indicate that NUH gives rise to higher coverage in contrast to UH. By employing RFA, the network performance improves in both cases but due to less interference the coverage in NUH is better than UH.In ζj and ζS/ζM = 10.Similarly, 2, NUH with RFA leads to 19% UL coverage improvement due to significant interference reduction.Both of these figures indicate that increasing IJs distribution area leads to improved coverage as the IJs become less effective. The figures further depict that at an area of 1 kmM. These figures consider ζj = 100 and ζS/ζM = 10. The results indicate that increase in the values of ΓM gives rise to lower coverage due to lower user connection. The figures also indicate that NUH with RFA and ΓM = -50 dB give rise to the highest coverage gain in contrast to the rest of the scenarios.Finally, M = −10 dB while using RFA and NUHs as compared with RFA and UHs. This work can be extended to evaluate drone-based jammers in HetNets.This work aims to reduce ICI and IJI by employing SBS muting and RFA in HetNets. Various network parameters such as jammer’s density, jammers transmit power and their distribution area, SIR threshold are investigated against user coverage. The results are obtained for both UHs and NUHs in addition to and without RFA. The results depict that NUHs employing RFA outperform other scenarios in terms of UL coverage. Moreover, the investigation indicates 20% UL coverage improvement at Γ