Conduct disorder is one of the most common developmental psychiatric disorders which is characterized by persistent aggressive and antisocial behaviors during childhood or adolescence. Previous neuroimaging studies have investigated the neural correlates underlying CD and demonstrated several constructive findings. However, Individuals with CD are at high risk for comorbidities, which might give rise to the inconsistencies of existed findings. It remains unclear which neuroanatomical abnormalities are specifically related to CD without comorbidities. Using structural magnetic resonance imaging (sMRI) data of 69 CD and 69 typically developing (TD) male youths (aged 14–17 years), the present study aims at investigating gray matter volume alterations of non-comorbid CD . We also examined how regional gray matter volumes were related to callous-unemotional (CU) traits and conduct problems in the CD group. The whole-brain analysis revealed decreased gray matter volumes in the right pre-postcentral cortex, supramarginal gyrus and right putamen in CD youths compared with TD youths. The region-of-interest analyses showed increased gray matter volumes in the superior temporal gyrus (STG) and right orbitofrontal cortex (OFC) in CD youths. Correlation analysis found that gray matter volume in the left amygdala was negatively correlated with CU traits in CD participants. These results demonstrated that gray matter volume in the orbitofrontal-paralimbic cortex, including OFC, STG and amygdala, might characterize the male youths with non-comorbid CD and might contribute to different severe forms and trajectories of CD. Conduct disorder (CD) is one of the most common developmental psychiatric disorders  which haNeural substrates abnormalities are important indicators of pathophysiological processes which can reflect disorder etiology . In recePrevious literature has indicated that these inconsistent findings might partially reflect the heterogeneity of the CD in the samples examined . It has In this context, accounting for the heterogeneity of CD youths would help improve the reliability of CD-sMRI study findings. A better understanding of the neural correlates underlying non-comorbid CD may, in turn, help clarify the etiology of CD. Although several previous researchers have studied the heterogeneity in individuals with CD, with promising findings, the existing sMRI literature has some important limitations, including relatively small sample size and large age range. Thus, the current study used sMRI in conjunction with voxel-based morphometry (VBM) to examine gray matter abnormalities in non-comorbid, non-medicated CD boys, in a relatively large sample size (69 CD and 69 TD boys) and within a small age range (14–17 years old). In line with previous studies, we expected to find gray matter volume alterations in several regions in the CD group, especially the regions involved in the ‘hot’ executive functions, such as OFC, STG and limbic brain regions . We alsoGiven that CD rates are greater in males and to remove gender as a confounding variable , this stThe inclusion period for CD and TD subjects was between November 2011 to August 2018 see , which hFor TD group recruitment, male students who matched the CD subjects’ ages were recruited through flyers in the local middle schools. Volunteers were interviewed by the same psychiatrists and subjected to the SCID-I/P. Information provided by TD subjects was verified by their parents on an as-needed basis. None of the TD participants met the criteria for CD. Moreover, the exclusion criteria for both groups were assessed in diagnostic interviews. The exclusion criteria included: history of ADHD, or any other behavioral disorder; history of any psychiatric or emotional disorder; the presence of any pervasive developmental or chronic neurological disorder , Tourette’s syndrome, post-traumatic stress disorder, obsessive-compulsive disorder; persistent headaches, head trauma; alcohol or SUD in the past year; contraindications to MRI; or a full-scale IQ < 80, as estimated by the Wechsler Intelligence Scale for Children-Chinese revision (C-WISC) examinations .The Chinese version of the Center for Epidemiologic Studies Depression Scale (CES-D)  and the T1-weighted three-dimensional magnetization-prepared rapid gradient-echo sequences with following acquisition parameters: voxel size = 1 × 1 × 1 mm3, repetition time = 8.5 ms, echo time = 3.743 ms, flip angle = 8°, acquisition matrix = 256 × 256 pixels, field of view = 256 × 256, number of slices = 180, slice thickness = 1 mm.Images were acquired by a Philips Achieva 3-T scanner  with a standard head coil at the Medical Imaging Department in Second Xiangya Hospital of Central South University . MRI data were acquired with http://www.neuro.uni-jena.de/cat/) provided ratings of image data quality, which were used to identify problems with individual images. These ratings assess basic image properties, noise and geometric distortions  and combine them into a weighted image quality rating (IQR). All of the scans rated B and above in IQR, representing good image quality. Finally, an automated quality check using covariance analysis on the sample homogeneity of segmented gray matter images was performed in CAT12 toolbox ; the anterior commissure was set manually in each image as the origin of the spatial coordinates. The Template-O-Matic toolbox was used to create a standardized a priori tissue probability map (TPM) based on the age of the 138 participants (Images were cropped and reoriented following the anterior-posterior commissure line with MRIcroicipants . The higicipants . Diffeomicipants . The temP < 0.05 was used with voxel-level familywise error (FWE) rate correction for multiple comparisons to protect against type I errors   . Prior Ron (SVC)  ROIs froon (SVC)  given thon (SVC) .P < 0.05, voxel-level FWE correction. Where significant results were found, contrast estimates from the peak voxel were extracted to assess the direction of effects in SPSS 19.0 and plot the results.Besides, we conducted exploratory whole-brain regression analysis between gray matter volume and CU traits in CD group only . Regression analysis was also used to investigate the relationship between conduct problem scores  with gray matter volumes in CD group. Results were thresholded at The demographic and clinical characteristics of the subjects are summarized in k = 37, P = 0.038, Z = 3.56, SVC) and increased left OFC volume  in the CD group when compared with TD group. We did not find any other significant gray matter volume differences between groups elsewhere in the brain  was negatively correlated with CU traits in CD participants . However, a positive correlation between gray matter volume in right OFC  and conduct problem scores was observed at a more liberal significance threshold (Puncorrected < 0.001). No other correlation was detected in the brain.The whole-brain analysis did not detect any brain regions that showed a correlation with conduct problem scores for SDQ in the CD group , the reduced striatal volume effect was non-significant when factoring out ADHD symptoms or excluding participants with comorbid ADHD. This result is consistent with studies showing that ADHD is associated with reduced volume in the putamen  in CD participants. Although we did not predict this finding, the result is consistent with previous studies on adolescents with conduct problems  and maleThe present study observed robust negative associations between CU traits and gray matter volumes in the left amygdala in CD participants, which is consistent with previous fMRI studies of CD demonstrating a negative correlation between CU traits and amygdala activity . CU traiWe noted several limitations of the present study. Firstly, the age-of-onset data were based on retrospective reports, which might be associated with recall bias. To minimize the effect of recall bias, we collected data from both participants and their caregivers. The psychiatrist would ask for more details and make the final judgment if the information provided was inconsistent. Secondly, the clinical characteristics of the subjects were based on self-perspective data. For CD participants with tendencies to manipulate, deceive or minimize faults, the validity of self-report assessment may be compromised . InconsiTo our knowledge, this is the largest single VBM study to investigate the gray matter volume alterations of CD boys with consideration of CU traits and age-of-onset. In conclusion, the present study replicated the gray matter volume aberrations in supramarginal gyrus, STG and OFC and extended these findings to male youths with non-comorbid CD. The results further suggested critical roles for the orbitofrontal-paralimbic cortex, including OFC, STG and amygdala in affecting empathy and moral judgment in ways that contribute to observable psychopathological symptoms of CD.The datasets generated for this study are available on request to the corresponding author.The studies involving human participants were reviewed and approved by the Ethics Committee of the Second Xiangya Hospital of Central South University (No. CSMC-2009S167). Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin.SY and XW: study concept and design. WS and SY: study supervision. YG, YJ, QM, JZ, DD, QW, and XG: acquisition, analysis, or interpretation of the data. YG: drafting of the manuscript. SY and WS: administrative, technical, or material support. All authors: critical revision of the manuscript for important intellectual content.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.


In vitro single-molecule methodologies have significantly aided the identification of links between TF gene dosage and transcriptional outcomes. Additionally, advances in quantitative microscopy have contributed mechanistic insights into normal and aberrant TF function. However, to understand TF biology, TF-chromatin interactions must be characterised in vivo, in a tissue-specific manner and in the context of both normal and altered TF numbers. Here, we summarise the advanced microscopy methodologies most frequently used to link TF abundance to function and dissect the molecular mechanisms underlying TF HIs. Increased application of advanced single-molecule and super-resolution microscopy modalities will improve our understanding of how TF HIs drive disease.Transcription factors (TFs) are life-sustaining and, therefore, the subject of intensive research. By regulating gene expression, TFs control a plethora of developmental and physiological processes, and their abnormal function commonly leads to various developmental defects and diseases in humans. Normal TF function often depends on gene dosage, which can be altered by copy-number variation or loss-of-function mutations. This explains why TF haploinsufficiency (HI) can lead to disease. Since aberrant TF numbers frequently result in pathogenic abnormalities of gene expression, quantitative analyses of TFs are a priority in the field.  Summary: Abundance and chromatin interactions determine transcription factor function; their disruption can impair development and cause disease. We survey advanced microscopy methodologies that measure these properties and discuss molecular mechanisms that underlie transcription factor haploinsufficiency. Precise tissue- and cell-specific regulation of gene expression is required for development and homeostasis. Transcription factors (TFs) tightly control transcriptional programmes in a cell- and tissue-dependent manner. General TFs and the core transcriptional machinery bind to gene promoters, whereas specific activating and repressing TFs bind to gene regulatory elements – i.e. transcriptional enhancers or silencers – and interact with promoter-bound complexes to control transcription . This inThe main TF families are classified according to their DNA-binding domains. These include the helix-turn-helix (HTH), zinc-finger (ZNF), basic helix-loop-helix (bHLH), basic leucine zipper (bZIP) and nuclear hormone receptor binding domains , each feThe aberrant function of TFs can have profound effects on development and disease. Of particular interest is how reduced levels of some TFs may influence their function and result in haploinsufficient phenotypes. Haploinsufficiency (HI) is the inability of a gene to rely on only one (of its two) alleles to exhibit its normal function. Therefore, HIs generally result in loss-of-function (LOF), often abnormal, phenotypes. It follows that HIs are properties of genes and, although ‘dosage-sensitive genes’ is a broader term than ‘HI genes’, henceforth, for simplicity, we use the two terms interchangeably.n≥0.9 as intolerant and with a score of n≤0.1 as tolerant to LOF mutations  , i.e. rautations . Bioinfoutations , linkingutations , keratitutations  and oculutations , as wellutations , schizenutations , craniosutations  and Greiutations . Diseaseutations  to ribosutations  and cancutations . Interesutations .Over 1600 human TF genes have been grouped into gene families , allowinabsolute TF numbers;stoichiometry of TFs and their interacting proteins;availability and binding kinetics to their TF target sites;stochastic nature of gene transcription;biophysical properties, such as their ability to form condensates.Insufficient TF protein levels are the common denominator of haploinsufficient TF genes. In these cases, the abnormal phenotypes are triggered in a dosage-dependent manner . At the How these parameters vary simultaneously between or within cells, tissues or entire organisms adds to the sheer complexity in characterising the causes of dosage sensitivity and HI in humans . HI may Studying the mechanisms of TF concentration and function in TF HIs is becoming increasingly possible thanks to the continuous advancement of microscopy methods. NANOG and GATA6 in a ‘salt-and-pepper’ pattern in the mouse embryonic inner cell mass controls the segregation of the epiblast from primitive endoderm lineages  analyses of human TFs found over a third of them to be enriched in specific tissues . Spatiotlineages . Stemneslineages . For inslineages . When Poelopment , 2005. TPOU5F1 enhancer is regulated by the accumulation of SOX2 and the chromatin reader BRD4, their chromatin-binding dynamics, as well as their interactions with the Mediator complex and the elongation controller P-TEFb, a cyclin-dependent kinase consisting of CDK9 and one of several cyclin subunits , a maternally provided TF and key morphogen, which determines embryonic anteroposterior polarity, remains unaltered despite its low levels within posterior nuclei. This is achieved when Bcd binds hubs together with the pioneer TF Zelda (Zld), which results in increased local concentrations  or super-resolution microscopy (SRM), respectively . FCS allelopment , the TF-elopment , TetR  and single-cell RNA sequencing (scRNA-seq). smFISH has been used to measure levels of FOXO mRNA  and heterozygous pathogenic mutations result in ocular developmental defects that are similar between mice  and SPI1 , bind ‘closed’ chromatin, making it accessible to non-pioneer TFs that act either alone or through interactions with chromatin remodelers   or bimolpression . TF comppression   gene guarantees the potentiation of sog transcription in the early the Drosophila embryo, such that all cells express the same amount of sog – even in positions where the Dl morphogen TF concentration is low  in combination with SPT (LLSM-SPT) , LLSM-FCand TIRF . Moreoveand TIRF  and 3D-Sand TIRF . HoweverPou5f1 and Nanog genes, to link clusters of BRD4 and enhancers with active transcription  microscopy and LLSM – is used to observe cellular localisation and co-partitioning of proteins into condensates , and canThe mobility of TFs within condensates, and the exchange of molecules between condensates and the environment, have been measured by FRAP, FCS and SPT. Condensates formed by transcriptional machinery components, such as BRD4 and MED1 , and TF It will be interesting to further investigate the biological functions of TF condensates through a combination of single-molecule approaches to link TF numbers and their dynamic behaviour to gene expression and HIs.Here, we have outlined how TF abundance and dynamic interactions with chromatin are required for normal development and how abnormalities in such regulation can result in disease.We have discussed the key factors and cellular functions that control TF abundance in cells and tissues. TFs undergo complex kinetic interactions with chromatin; therefore, investigating TF numbers and molecular movement at the cellular and tissue levels is essential. Studied paradigms of dosage-sensitivity and HI emphasise how recent quantitative microscopy advances help researchers to investigate such complex TF interactions and their malfunction in disease. To date, such information on TFs and TF-cofactor complexes can be obtained with high spatiotemporal resolution. When these methodologies are applied to multiple cell types and disease models, they help to understand the molecular underpinnings of HI-associated diseases. However, a remaining challenge is to simultaneously study the behaviour of collaborating TFs and TF-complexes in live cells and tissues. This shortcoming is likely to result from the limitations of fluorescence microscopy to faithfully investigate several differently labelled proteins at the same time. Mass spectroscopy, RNA-seq and ChIP-seq methods do provide much broader, albeit static, datasets of TF binding and protein and mRNA abundances in cells but lack the dynamic information of live systems. A further challenge is to combine temporal and spatial super-resolution methods so that the mobility and chromatin-binding dynamics of individual TFs can be investigated at specific genomic loci. As discussed, this is currently limited to repetitive loci, which can be studied in tandem using the same fluorescent probe. Non-repetitive DNA sequences require a large number of sequence-specific probes to sufficiently increase the signal-to-noise ratio for fluorescence imaging .YAP1 (Nevertheless, when studying the function of fluorescently labelled TFs, microscopy methodologies have become sensitive enough to detect molecules at low, physiologically relevant concentrations and by using low excitation power, which largely preserves the normal cell function. In fact, smaller and brighter fluorescent tags have been identified  but taggYAP1 . The genin vitro quantification of TF transcriptomes and interactomes. To date, most studies on TF function have addressed the impact of their mutations in disease only qualitatively. The information on intracellular TF concentrations and chromatin-binding behaviour is scarce, and may have been underappreciated. Until now, computational and bioinformatics analyses, coupled to machine learning and high-throughput screening, have been predominantly deployed to predict TF binding sites (From a translational perspective, new animal models will enable the detection of early developmental transcriptional and signalling defects that normally lead to disease onset and progression. This will be possible by combining high spatial and temporal resolution microscopy – as discussed in this Review – with ng sites  or TF-bing sites . Further


Long non-coding RNAs (lncRNAs) play important roles in the pathogenesis of Alzheimer's disease (AD). However, the functions and regulatory mechanisms of lncRNA are largely unclear. Herein, we obtained 3,158 lncRNAs by microarray re-annotation. A global network of competing endogenous RNAs (ceRNAs) was developed for AD and normal samples were based on the gene expressions profiles. A total of 255 AD-deficient messenger RNA (mRNA)-lncRNAs were identified by the expression correlation analysis. Genes in the dysregulated ceRNAs were found to be mainly enriched in transcription factors and micro RNAs (miRNAs). Analysis of the disordered miRNA in the lncRNA-mRNA network revealed that 40 pairs of lncRNA shared more than one disordered miRNA. Among them, nine lncRNAs were closely associated with AD, Parkinson's disease, and other neurodegenerative diseases. Of note, five lncRNAs were found to be potential biomarkers for AD. Real-time quantitative reverse transcription PCR (qRT-PCR) assay revealed that PART1 was downregulated, while SNHG14 was upregulated in AD serum samples when compared to normal samples. This study elucidates the role of lncRNAs in the pathogenesis of AD and presents new lncRNAs that can be exploited to design diagnostic and therapeutic agents for AD. Alzheimer's disease (AD) is the most common cause of dementia and is characterized by age-dependent memory loss and impairment of multiple cognitive functions  are small, endogenous, non-coding RNA (ncRNA) molecules that play important regulatory roles in AD  are a major type of ncRNAs that can regulate genes at the transcriptional, post-transcriptional, and epigenetic levels  database. The mRNA, miRNA, and lncRNA expression profiles were obtained through chip re-annotation, data integration, and standardization. The miRNA-mRNA and miRNA-lncRNA interaction data were obtained from starBase database. The hypergeometric distribution approach was used to establish a background ceRNA regulation network. Expression correlation was used for pruning after which functional module mining was performed through a co-expression network to identify candidate diagnostic biomarkers or potential therapeutic targets.GSE133349). We matched the probes with their corresponding genes. In cases where multiple probes corresponded to one gene, the median was expressed. Where one probe corresponded to multiple genes, such a probe was excluded.We searched the GEO database and the Affymetrix Human Genome U133 Plus 2.0 Array chip platform to identify gene expression profile data using the keyword “Alzheimer's disease.” Samples sizes with ≥30 datasets and 7 datasets were obtained. In addition, the dataset GSE16759, which simultaneously detected the expression profiles of miRNA and mRNA, included eight sets of 629 samples. Forty samples of undefined AD were excluded, and finally 589 samples were included as shown in To obtain the latest lncRNA expression profile, lncRNA re-annotation was performed using the HG-U133 2.0 array. First, the GENCODE Release 21 version of lncRNA transcript sequences was downloaded from GENCODE. Then, probe clusters sequences were aligned to lncRNA sequences using seqmap  of Pearson's correlation coefficient difference was applied to identify dysregulated mRNA-lncRNA.We compared the global networks of ceRNAs in normal and AD samples and selected the intersection of two networks. Furthermore, the difference between the Pearson's correlation coefficients of mRNA-lncRNA in two ceRNA networks was calculated. With 1,000 random readings as the statistical background, the probability (Each sample was subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of ssGSEA using the R package GSVA38 to identify pathways associated with lncRNA expression, applying a correlation > 0.5.https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi), with the ID: GSE133349.To facilitate the reproduction of our results, the eight datasets were uploaded to the GEO database  was used to extract total RNA from serum samples. The synthesis of total RNA into cDNA was performed according to the instructions of the Reverse Transcription Kit . The ABI 7500 real-time PCR instrument was used to perform the real-time PCR reaction using the quant one-step real-time quantitative reverse transcription PCR (qRT-PCR) Kit . The primers used were as follows: SNHG14 forward, ATGAGCTGACAACCTACTCC and reverse, AAGTCATCTTCTGCAAGGGT; PART1 forward, CCCTTTCACTATGAAGGACC and reverse, ATTTACCCGTCCAGTTCTG; NNT-AS1 forward, AGAAACAGGTCTAAAGACCCT and reverse, TTCTTGGCATCTCTGAGCA; AC093010.3 forward, TGATGTGTTTGCTATCTGCT and reverse, TGTTAACAGCTAGCCATTCAG; ARMCX5-GPRASP2 forward, AAGAGAAGGGATAGAGTGGTG and reverse, CTTCTGTCATAGAAATTTCCCTCTC; GAPDH forward, TCAAGATCATCAGCAATGCC and reverse, CGATACCAAAGTTGTCATGGA; GAPDH were used as internal controls. 2t-test, correlations were determined using Pearson's correlation coefficient, g:profiler  was used for network visualization.Statistical analyses were performed using R 3.4.3, and all analyses (non-specific description) were performed with default parameters. Group comparisons were performed using the Student's To investigate the potential role of miRNA-mediated ceRNA networks in AD, we analyzed the ceRNA landscape in normal samples and AD samples based on the expression profiles . Using tTo explore the co-expression relationship of the mRNA-lncRNAs in the ceRNA network, we analyzed the relationship between network degree and mRNA-lncRNA co-expression. The network degree was significantly positively correlated with mRNA-lncRNA co-expression . Genes wp ≤ 0.05 were selected as the background of the random network. A total of 294 dysregulated mRNAs-lncRNAs were identified in training queue, and a total of 333 dysregulated mRNA-lncRNAs were identified in the verification cohort by the same method. In addition, 258 dysregulated mRNA-lncRNAs were selected from all samples by the same method. By comparing the dysregulated mRNA-lncRNAs in the three cohorts, a high degree of overlap was found between them  ceRNAs were unchanged while 3,696 (58.9%) ceRNAs disappeared from the AD samples . Analysieen them . This sheen them . NotablyTo elucidate on the functional implications of the ceRNA network, functional enrichment analysis was performed on all 231 genes in the ceRNA network. Results showed that these genes were enriched in two gene ontology (GO) terms and were not associated with any pathway , implyinp < 0.05 as the cut-off value for selection, 86 pairs of miRNA-lncRNA and 111 pairs of mRNA-miRNA, including 19 miRNAs, 131 genes, and 37 lncRNAs, were identified. Interestingly, unlike mRNA-lncRNA, these dysregulated miRNA-target pairs were downregulated in AD samples  lncRNAs to be significantly differentially expressed . SimilarTo investigate the diagnostic value of lncRNA in AD, a receiver operating characteristic (ROC) curve classification was performed for the nine lncRNAs in AD samples and normal samples. Results showed that each of the nine lncRNAs could diagnose AD with an average area under the curve (AUC) of 0.6 . HoweverAdvances in high-throughput experimental techniques have enabled a comprehensive analysis of ceRNA interactions using the gene expression correlation approach. In addition, public databases and computational methods such as starBase provide platforms for the analysis of CLIP-seq-supported miRNA-mRNA interactions  can be found in the article/The studies involving human participants were reviewed and approved by the Neuroscience Center, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University. The patients/participants provided their written informed consent to participate in this study.DX and CH conceived and supervised the research. SD analyzed the data. JX wrote the manuscript. JL and NL prepared figures and tables. DX identified the research and editorial manuscript. All authors read and approved the manuscript.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.


At follow-up, participants with low food literacy at the program end significantly improved their follow-up domain scores for plan and manage (60%) and selection (73.3%), and participants with moderate or high food literacy at the program end maintained their follow-up scores. A food literacy program can support adults to improve and maintain their food literacy behaviours and maintain dietary behaviour change; therefore, strategies to support this continued change must be considered.Food Sensations for Adults is a free four-week nutrition and cooking program that teaches low- to middle-income individuals food literacy. This research aimed to compare demographic characteristics of participants who completed the program’s follow-up questionnaire three months after program completion and assess whether food literacy and dietary behaviour changes were improved or maintained. Statistical analysis methods used factor scores of the plan and manage, selection, and preparation domains to examine mean self-reported changes in food literacy. Tertile stratification methods calculated changes in participants who had low, middle, and high end-of-program food literacy scores, and multivariable regression analysis explored the associations. The follow-up results ( Food literacy is ‘composed of a collection of inter-related knowledge, skills, and behaviours required to plan, manage, select, prepare, and eat food to meet needs and determine intake’ . The fouBest practices for providing low-income audiences with effective nutrition education indicate that conducting outcome (follow-up) evaluation and assessing maintained behaviour change are two critical evaluation practices . A curreThere are only two Australian food literacy programs that have published follow-up evaluation findings ,9,10. ThSince 2007, Foodbank WA has delivered a food literacy program across WA , with thThis research aims to determine whether participants continue to improve or maintain food literacy behaviours and selected dietary practices three months after the program’s end. The program’s long-term influence is established by assessing participants’ food literacy and dietary behaviours between the end-of-program and follow-up questionnaires, and analysing factors that associate with continuing improvements or maintained improvements in food literacy behaviours between the beginning and end of the program.Cross-sectional questionnaires were conducted on three occasions—the program’s beginning and end, and three months after completion. Participants completed hard copy questionnaires at the beginning and end of the program and non-attendees completed the questionnaire online. The end-of-program questionnaire asked participants to indicate how they would like to complete their voluntary follow-up questionnaire: online, posted hard copy, or telephone. The participants were not directly paid for completing the follow-up questionnaire; rather, they were automatically entered into a draw every two months to win a A$200 supermarket voucher. Not all programs were evaluated due to literacy levels in ethnically diverse groups or confidentiality issues such as prison groups and not all participants consented to the evaluation. During the time period for this study, 2,841 participants attended one or more FSA sessions.Within the first questionnaire, participants completed a 14-item food literacy behaviour checklist and answered questions regarding their food literacy-related practices, selected dietary behaviours, and socio-demographic information. The end-of-program questionnaire included the same 14-item food literacy behaviour checklist and questions regarding selected dietary behaviours, and the follow-up questionnaire included the same 14-item food literacy behaviour checklist and questions regarding food literacy-related practices, selected dietary behaviours with an additional question on the barriers to change experienced.Within each of the questionnaires, different sets of questions were asked. The first included questions regarding eight socio-demographic characteristics: sex, age group, household composition, education level, employment status, socio-economic index, whether they were born in Australia and whether they were Aboriginal or Torres Strait Islanders. All questionnaires included two questions regarding self-reported dietary behaviours—self-reported intake of fruit and vegetables (serves per day) and the consumption frequency of fast food meals and sugar-sweetened drinks. Dietary behaviour questions included images of serve sizes for fruit and vegetables and examples of fast food meals and sugar-sweetened drinks from the Australian Guide to Healthy Eating to guide participants . Questiop-value < 0.05 was considered statistically significant. Fruit and vegetable intake was treated as a continuous variable, and all other variables were treated as categorical.The statistical analysis was completed using SPSS Statistics version 25 . The A participant’s demographic characteristics—sex, age group, household composition, education level, employment status, socio-economic index, whether they were born in Australia, and whether they were Aboriginal or Torres Strait Islanders—were used to identify participants less likely to complete the follow-up questionnaire. Chi-square tests were used to compare the demographic information of attendees who completed both the end-of-program and follow-up questionnaire against those who did not complete the follow-up questionnaire.The food literacy behaviour questions were used as the foundation for three food literacy behaviour domains in each questionnaire: plan and manage, selection and preparation . The facTo examine food literacy behaviour changes between the questionnaires, an innovative statistical approach was applied. The food literacy behaviour score was first allocated between three quantiles or tertiles. In clinical and epidemiological studies, quantiles are used in an analysis to separate distribution of data into groups and identify those more at risk of specific outcomes ; when spMcNemar–Bowker tests were used to investigate whether food literacy behaviour factor scores changed or remained the same between the end-of-program and follow-up questionnaires. The tests compared the tertile ranking of participants at these stages for the three food literacy behaviour domains—plan and manage, selection, and preparation. Paired t-tests were used to determine whether food literacy behaviours and dietary intake adjustments were maintained after the program’s completion. The three food literacy behaviour domains and the change in self-reported dietary intake of fruit and vegetable serves were reviewed in both the end-of-program and follow-up questionnaire scores. McNemar–Bowker tests were used to compare questionnaire scores of these two stages regarding self-reported intake of fast food meals and sugar-sweetened drinks and used to examine whether the program had improved cooking skills by comparing a participant’s self-reported cooking skills between their first and follow-up questionnaires.p-value indicates an increased likelihood.Multivariable regression analyses used a forced entry method to identify demographic(s), dietary behaviour(s) or food literacy-related practice(s) of participants who maintained or improved their moderate or high food literacy behaviour score between the end-of-program and follow-up questionnaires. Demographic characteristics included sex, age group, household composition, education level, employment status, socio-economic index, whether they were born in Australia and, whether they were Aboriginal or Torres Strait Islanders. Dietary behaviours included self-reported intake of fruit, vegetables, fast food meals, and sugar-sweetened drinks. Food literacy-related practices included the attitude towards the cost of healthy food, self-reported cooking skills, and the responsibility for preparing meals and purchasing food. Three food literacy behaviour domains—plan and manage, selection, and preparation—were analysed separately. Participants who scored moderately in the end-of-program questionnaire and moderate or high in the follow-up questionnaire, and those who scored high in both the end-of-program and follow-up questionnaires, were compared to all other participants (who scored low in both questionnaires or those who scored lower in the follow-up questionnaire). The results were displayed as an odds ratio, in which the likelihood of reporting a particular demographic or characteristic for participants who maintained or improved a moderate to high score compared to the reference answer (indicated by 1). An odds ratio greater than 1 with a significant Human Research Ethics Committee approval was obtained from Curtin University (RDHS-52-16). At the beginning of the program, each participant provided written consent to participate in the evaluation on an attendance sheet. If a participant consented to receive the follow-up questionnaire, they provided their contact details on their completed post-program questionnaire.n = 534 (86.8%) completed all three questionnaire stages and n = 594 (95.6%) completed the end-of-program and follow-up questionnaires. Completion methods of the follow-up questionnaire included online (75.4%), telephone (19.2%), and postage of a hard copy (5.5%).Out of 1,855 participants consenting to the evaluation processes, 621 (33.4%) completed the follow-up survey and the remainder either did not consent to follow-up, did not respond when contacted at the three-month follow-up, or withdrew consent at follow-up. Of the participants who completed the follow-up, There were several distinct demographic characteristics between participants who completed the follow-up questionnaire compared to those who did not. Those who completed the follow-up questionnaire were more likely to be female (84.2% vs. 78.4%), older (17.9% vs. 11.3% [66 and over]), living as a couple with children (39.2% vs. 33.5%) or without children (21% vs. 16.5%), and have a higher education level  see . Those wThe mean scores for the plan and manage and selection behaviours increased in the follow-up questionnaire compared to the participant’s end-of-program questionnaire score . The prep < 0.0001), the consumption frequency overall improved from the beginning of the program (p < 0.0001). There was no change in the reported intake frequency of sugar-sweetened drinks between the end-of-program and follow-up questionnaires (p = 0.063), or from the beginning of the program to follow-up (p = 1). However, the consumption frequency of fast food meals and sugar-sweetened drinks overall was low before the program—28.6% of participants had never consumed fast food, and 47.9% had never consumed sugar-sweetened drinks. Participants’ self-reported cooking skills were measured twice, with self-reported cooking skills significantly improving between the program’s first and follow-up questionnaires , 33.3% reported difficulties or obstacles in establishing food-related changes at home, 25.9% reported taste preferences of household members, and 23.2% reported busy lifestyles to be barriers. Other barriers that participants experienced also included not wanting to change the intake of preferred foods (14.2%) and the price of healthy foods (13.2%).Participants were asked to report on any barriers or challenges they had experienced in the three-month follow-up questionnaire. Of those who answered the question, ‘Have you experienced any difficulties or obstacles making food-related changes at home?’ . Further research is required to understand the barriers to increasing healthy food intake when participants are reporting improving or sustaining food literacy behaviours. Many programs have documented increases in fruit and vegetable consumption; however, they may not have included measures of dietary behaviours or intake in the follow-up evaluation or included a follow-up evaluation at all ,3. The Un = 1605) of participants rated themselves as being able to cook almost anything or cook a wide variety of meals; at the follow-up, participants improved the perception of their skills [Participants’ perceptions of their preparation and cooking skills are associated with the frequency of all food literacy behaviours. The preparation and cooking domain of food literacy appears to be the active ingredient for individuals using food literacy to improve their dietary intake. Fredericks et al. discussed the ten experiential features of culinary or cooking programs that instigate behavioural changes based on the experience of nutrition educators . At the r skills . A food r skills  and provr skills .Food literacy is a complex set of knowledge, skills and behaviours. Therefore, it is a complex process to transform knowledge and skills into positive behaviours successfully, and this change requires further research. Food literacy programs tend to attempt to change cognitions, such as attitudes and apply constructs from social cognitive models, such as improving self-efficacy . ApplyinFurther research is required to understand the cues for food literacy behaviour change and the reward received when making changes that assist in creating habits. Opportunities for specific participants to attend advanced programs or refresher programs may assist in maintaining and improving behaviour change. In addition, technology support applications and programs could provide reminders regarding using food literacy to improve fruit and vegetable intake and reduce fast food meal intake ,33. PartThere are numerous limitations to acknowledge. First, the ideal follow-up or outcome evaluation time is unknown. At the three-month evaluation point, results may have missed potential changes in the preparation domain. Second, community-focused programs present challenges for evaluation because, unlike research trials, they do not contain a control group. It is also possible that participants may engage other strategies post-program to continue improvements; therefore, not all change can be attributed to the program. Third, the demographic characteristics of those completing the follow-up questionnaire are different. It may be the highly motivated participants that complete the questionnaire, which potentially introduces a retention bias. Fourth, the evaluation included several short self-reported dietary behaviour questions. It is necessary to deepen the understanding regarding the association between dietary behaviour change using comprehensive measures and food literacy behaviours, and the effect of barriers to using food literacy behaviours post-program to improve dietary intake. Lastly, all program evaluation data is reliant on self-reported responses and is, therefore, subject to social desirability bias, whereby participants respond in a socially acceptable way as opposed to reporting their actual behaviour. Participation in Food Sensations for Adults resulted in the more frequent use of plan and manage, selection, and  preparation skills in the longer term. Food literacy programs are effective; however, there is a need to continue assisting and supporting participants to implement their skills—particularly food preparation—to maintain changes. Further investigation strategies for supporting and maintaining behaviour change after program completion is required.


The aim of the present study is to compare the clinical outcomes and postoperative complications of lumbar endoscopic unilateral laminotomy bilateral decompression (LE-ULBD) and minimally invasive surgery transforaminal lumbar interbody fusion (MIS-TLIF) to treat one-level lumbar spinal stenosis (LSS) without degenerative spondylolisthesis or deformity.A retrospective analysis of 112 consecutive patients of one-level LSS undergoing either LE-ULBD or MIS-TLIF was performed. Patient demographics, operation time, estimated blood loss, time to ambulation, length of hospitalization, intraoperative and postoperative complications were recorded. The visual analog scale (VAS) score for leg and back pain, the Oswestry Disability Index (ODI) score, and the Macnab criteria were used to evaluate the clinical outcomes. The healthcare cost was also recorded.The operation time, estimated blood loss, time to ambulation and length of hospitalization of LE-ULBD group were shorter than MIS-TLIF group. The postoperative mean VAS and ODI scores decreased significantly in both groups. According to the modified Macnab criteria, the outcomes rated as excellent/good rate were 90.6 and 93.8% in the two groups. The mean VAS scores, ODI scores and outcomes of the modified Macnab criteria of both groups were of no significant difference. The healthcare cost of LE-ULBD group was lower than MIS-TLIF group. Two cases of intraoperative epineurium injury were observed in the LE-ULBD group. One case of cauda equina injury was observed in the LE-ULBD group. No nerve injury, dural injury or cauda equina syndrome was observed in MIS-TLIF group. However, one case with transient urinary retention, one case with pleural effusion, one case with incision infection and one case with implant dislodgement were observed in MIS-TLIF group.Both LE-ULBD and MIS-TLIF are safe and effective to treat one-level LSS without degenerative spondylolisthesis or deformity. LE-ULBD is a more minimally invasive option and of less economic burden compared with MIS-TLIF. Decompression plus instrumented fusion may be not necessary for one-level LSS without degenerative spondylolisthesis or deformity. Lumbar spinal stenosis (LSS) can be caused by degenerative facet joints, hypertrophic ligamentum flavum, bulging or protrusion of the intervertebral discs, spondylolisthesis and or a combination of the above pathological lesions , 2. ThesMicrosurgical procedures was firstly described by Yasargil and Caspar in 1977 . MicroscLaminotomy with medial facetectomy may result in segmental spinal instability, instrumented fusion may be necessary in some cases to prevent segmental spinal instability after decompression of LSS , 13. TraEven though both decompression alone and decompression plus instrumented fusion has been performed to treat patients with LSS, it is controversial about the necessity of instrumented fusion after decompression , 18, 19.This retrospective study included 112 patients  with one-level LSS, who underwent LE-ULBD or MIS-TLIF in our department between January 2016 and December 2017. The present study was conducted in accordance with the guidelines of the Declaration of Helsinki and was approved by the ethics committee of our College. Written informed consents was obtained from each patient. In these patients, 32 surgeries were performed by LE-ULBD and the other 80 surgeries were performed by MIS-TLIF.Inclusion criteria were as follows: patients with (1) main symptoms, including leg pain, numbness, motor weakness or neurogenic claudication; (2) computed tomography and/or magnetic resonance imaging indicating central stenosis and/or lateral recess stenosis, in agreement with clinical symptoms and signs; (3) a history of failed conservative treatment, including physical therapy and nonsteroidal anti-inflammatory drugs intervention for more than 3 months; and 4) follow-up for at least 24 months. Exclusion criteria were as follows: patients with (1) degenerative spondylolisthesis or deformity; (2) tumors, infections, or other lesions; and (3) a surgical history involving the corresponding segment  follow-u. Each paAll the surgeries were performed by the senior author, who has many years of experience in open and minimally invasive lumbar canal decompression, and lumbar endoscopic visualized discectomy. Both LE-ULBD and MIS-TLIF were performed under general anesthesia with the patient in the prone position.Posteroanterior and lateral fluoroscopy were used to locate the interlaminar space at the surgery segment. A 10 mm skin incision was made laterally to the outer border of the interlaminar window. Soft tissue expanders were applied via the incision in order to separate the muscles to allow the insertion of the working sheath and the endoscopic surgical system . All the subsequent procedures were performed under constant irrigation with excellent endoscopic visualization.The inferior edge of the cranial lamina and the base of the spinous process of the ipsilateral side were removed by the endoscopic burr, enabling access into the spinal canal. Once the epidural space was entered, undercutting of the contralateral cranial lamina was performed. Then the ipsilateral and contralateral ligamentum flavum was identified and removed piecemeal with endoscopic punches and forceps. Subsequently, ipsilateral and contralateral medial facetectomy was performed to decompress the lateral recess and foramen, ensuring adequate decompression of the traversing nerve root. Once the traversing nerve root was decompressed, it was reflected medially using a blunt dissector. These procedures were presented in Fig. Posteroanterior and lateral fluoroscopy were used to locate the pedicles of the surgery level. The Wiltse approach was undertaken through a paramedian skin incision. A Quadrant tubular dilator was used for unilateral facet exposure. Facetectomy was performed on the ipsilateral side in order to visualize the transforaminal disc space. Laminectomy and lateral recess decompression were performed to decompress the spinal canal. Besides, the tubular retractor could be angled medially to complete a more extensive decompression of central canal stenosis and the contralateral side . LigamenThe operation time, estimated blood loss, time to ambulation, length of hospitalization, Intraoperative and postoperative complications were recorded. Follow-up examinations were conducted at 3, 6, 12 and 24 months postoperatively. Postoperative magnetic resonance imaging or computed tomography imaging was performed when necessary. The visual analog scale (VAS) score for leg pain and back pain , Oswestry Disability Index (ODI) score  and modified Macnab criteria were recorded preoperatively and at follow-up postoperatively. The surgery cost, anesthesia cost, surgical equipment and medical materials cost was also recorded.p-value of less than 0.05 was considered statistically significant.All data are presented as mean ± standard deviation. SPSS 22.0  was used to perform the statistical analyses. GraphPad Prism 6  was used to generate plots. Nonparametric statistical analysis, including Mann-Whitney U test or Wilcoxon signed-rank test were used. A The clinical characteristics of both groups were summarized in Table The operation time, estimated blood loss, time to ambulation and length of hospitalization of LE-ULBD group were shorter than MIS-TLIF group Table . The meaIntraoperative and postoperative complications of LE-ULBD and MIS-TLIF group were also compared Table . Two casThe surgery cost, anesthesia cost, surgical equipment and medical materials cost of LE-ULBD group were less than MIS-TLIF group Table .Table 6LSS is traditionally treated with open or microscopic laminotomy and foraminotomy via a midline lumbar incision . HoweverDuring LE-ULBD, laminotomy and and foraminotomy could be safely performed under excellent endoscopic visualization to guarantee complete decompression, minimize surgical trauma, and prevent spinal instability , 10, 11.The advantages of LE-ULBD were to perform bilateral decompression via a unilateral approach with minimize traumatization to the paraspinal musculoligamentous structures; to ensure the sufficient decompression the lateral recess and foramen under excellent endoscopic visualization to minimize neurological injury; and to preserve the stability of the spine with minimized foraminotomy , 10, 11.MIS-TLIF has been demonstrated to be a safe option for lumbar fusion with minimized iatrogenic traumatization to the paraspinal musculoligamentous structures , 17. ComWhile it is controversial about the necessity of instrumented fusion after decompression of LSS , 18, 19.There are some limitations to the present study. First, it is a retrospective, non-randomized controlled cohort study with a small sample size and short follow-up period. There also may be selection bias, as surgeons determined whether decompression alone or decompress with instrumented fusion should be performed. Further prospective, randomized, controlled studies, with larger sample sizes and longer follow-up periods should be conducted to determine the optimal surgical management for patients with LSS. Second, all the patients included were of one-level LSS without degenerative spondylolisthesis or deformity. As a result, further studies should be conducted to compare the clinical outcomes of LE-ULBD and MIS-TLIF for LSS with degenerative spondylolisthesis or deformity. Besides, preexisting adjacent level degeneration was not evaluated and compared in the present study.Both LE-ULBD and MIS-TLIF are safe and effective to treat one-level LSS without degenerative spondylolisthesis or deformity. LE-ULBD was proved to be a more minimally option and of less economic burden compared with MIS-TLIF. Decompression with instrumented fusion may be not necessary for one-level LSS without degenerative spondylolisthesis or deformity.


Up to 50% of myocarditis events developed in cancer patients upon treatment with immune checkpoint inhibitors (ICIs) are fatal. Therefore, identification of clinical risk factors predicting myocarditis onset during treatment with ICIs is important for the purpose of cardiac surveillance of high-risk patients. The aim of this retrospective matched case-control study was to assess whether pre-existing cardiovascular conditions were associated with the reporting of myocarditis with ICIs in VigiBase, the World Health Organization global database of suspected adverse drug reactions. Taking drugs labelled for the treatment of cardiovascular conditions as a proxy for concomitant cardiovascular risk factors and/or cardiovascular diseases, we found an association of moderate size between pre-existing cardiovascular conditions and the reporting of myocarditis with ICIs. Future prospective pharmacoepidemiological studies should assess the causal relationship between pre-existing cardiovascular conditions and myocarditis onset in a cohort of cancer patients followed during treatment with ICIs.p = 0.026, Cramer’s coefficient of effect size: Φ = 0.214). No significant association was found between pre-existing diabetes and ICI-related myocarditis reporting . These findings offer an invitation for future prospective pharmacoepidemiological studies to assess the causal relationship between pre-existing CV conditions and myocarditis onset in a cohort of cancer patients followed during ICI treatment.Although rare, immune checkpoint inhibitor (ICI)-related myocarditis can be life-threatening, even fatal. In view of increased ICI prescription, identification of clinical risk factors for ICI-related myocarditis is of primary importance. This study aimed to assess whether pre-existing cardiovascular (CV) patient conditions are associated with the reporting of ICI-related myocarditis in VigiBase, the WHO global database of suspected adverse drug reactions (ADRs). In a (retrospective) matched case-control study, 108 cases of ICI-related myocarditis and 108 controls of ICI-related ADRs other than myocarditis were selected from VigiBase. Drugs labeled as treatment for CV conditions (used as a proxy for concomitant CV risk factors and/or CV diseases) were found to be associated more strongly with the reporting of ICI-related myocarditis than with other ICI-related ADRs (McNemar’s chi-square test of marginal homogeneity:  Although rare, immune checkpoint inhibitor (ICI)-related myocarditis can be life threatening with a mortality rate up to 50% ,2. ICI-rTo date, data on clinical risk factors for ICI-related myocarditis are limited and heterogeneous. From a multicenter registry, amongst 35 patients with ICI-related myocarditis, 34.3% had pre-existing diabetes and 42.9% had an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker as pre-ICI home cardiovascular (CV) medication . Two disOur study aimed to assess whether pre-existing CV risk factors and/or CV diseases are associated with the reporting of ICI-related myocarditis in VigiBase. Using the description recorded in VigiBase of the role played by individual drugs in the occurrence of reported ADRs, concomitant drugs for CV conditions were taken as a proxy for the presence of pre-ICI CV risk factors and/or CV diseases. A (retrospective) matched case-control design (with the pre-existence of CV conditions as the independent variable and the reporting of ICI-related myocarditis as the dependent variable) was then adopted for comparing safety reports of ICI-related myocarditis, to control safety reports of ICI-related ADR(s) other than myocarditis from VigiBase.http://www.vigiaccess.org/) is the WHO global database of suspected ADRs, containing over 20 million safety reports submitted by physicians, pharmacists, healthcare professionals, patients/consumers, and pharmaceutical companies. To guarantee data quality, VigiBase employs a medical classification system , by which information on suspected ADRs is organized in a uniform and structured hierarchical form. The system organ classes (SOCs) serve to group ADRs by their manifestation site or etiology, and preferred terms (PTs) are used to describe ADRs by single medical concepts. To support retrieval of MedDRA-coded information, VigiBase offers the Standardized MedDRA Queries (SMQs) tool, which encompasses prespecified sets of PTs. Drugs are identified either by their active ingredients through the WHO Drug Global dictionary [VigiBase , patient characteristics (age and sex), ADR details (reported terms and outcomes), and drug descriptions  . Safety Deduplicated safety reports gathered in VigiBase up to 17 November 2019, were retrieved, from which safety reports with ipilimumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, and cemiplimab (active ingredients) as suspected drugs, were selected. A multistep selection process was then used to define the group of safety reports eligible for the study . Safety reports entered by physicians, pharmacists, and healthcare professionals, and classified by reporters as serious were included; in view of the study design, safety reports missing information on patient sex and/or age were excluded. Safety reports with suspected ICI(s) pertaining to patients on ICI treatment because of melanoma or non-small cell lung cancer (NSCLC), the two main indications of ICIs, were selected in the next step . SubsequCases were defined as the safety reports of ICI-related myocarditis or autoimmune myocarditis (MedDRA PTs) from among the group of safety reports resulting from the selection process. Of the remaining group of non-cases, 1709 safety reports were excluded for indicating ICI-related ADR(s) at the CV system or ICI-related ADR(s) representing risk factors for myocarditis , and/or for indicating an ICI-related transplant, immunosuppression, infection in an immunocompromised host, or autoimmune disorder . The ratAccording to the Human Research Act , from the Federal Assembly of the Swiss Confederation, ethical approval was not required for use of safety reports retrieved from VigiBase .A (retrospective) matched case-control study was conducted, with cases as described above and controls taken from the overall control group. Patient sex, patient age, cancer type, and ICI treatment , were used as matching criteria. The matching was done as random one-to-one matching, associating each case with a single control. The independent variable for the study was pre-existence of CV risk factors and/or CV diseases. Pre-existence of CV conditions was assessed using concomitant drugs of the anatomical, chemical, and therapeutic classes for the CV system (ATC C), for diabetes (ATC A10), and for obesity (ATC A08), as proxies. The dependent variable of the study was ICI-related myocarditis, that is, whether a safety report was a case or a control within the study.Data processing for the study included data validation, a matching procedure, and descriptive analysis of the selected group of safety reports, and was implemented in Python (version 3.8.2). From the safety reports in the case and control groups respectively, various demographic and clinical characteristics were summarized by counts and proportions. For establishing the relative risk (RR), per characteristic, for the case group compared to the control group, an odds-ratio (OR) approximation was employed to account for the size of the control group being imposed by the design of the study . An RR lFor analyzing the association between ICI-related myocarditis and pre-existing CV risk factors and/or CV diseases in the study group of safety reports, McNemar’s chi-square test of marginal homogeneity was employed. This non-parametric test for paired categorical data was applied to a 2 × 2 table tabulating all case-control pairs under study, and focused on the discordant pairs for testing whether the two marginal probabilities per category of the independent variable are the same; by disregarding the concordant pairs, the test was less affected by the matched design of the study than the standard chi-square test. Under the null hypothesis of marginal homogeneity, McNemar’s test statistic has a chi-squared distribution with a single degree of freedom. For evaluating the null hypothesis, a (two-tailed) significance level of α = 0.10 was used to allow for a reasonable power of the study in view of a small study-group size. Upon rejection of the null hypothesis, Cramer’s Φ coefficient of effect size was employed as a measure of strength for the association found between the independent and dependent variables.https://www.graphpad.com/quickcalcs/).All statistical tests were performed using the public web-based tool GraphPad .p = 0.026). Using Cramer’s Φ coefficient of effect size, the observed deviation from marginal homogeneity was found to be an association of moderate size (Φ = 0.214) between the independent and dependent variables [For the 108 case-control pairs in the study, ariables .p = 0.752). The overall study group of cases and controls did not include any safety report mentioning the use of antiobesity preparations (ATC class A08) and therefore did not allow an analysis of their potential association with the independent variable.In view of the 108 case-control pairs of our study, McNemar testing at the preset significance level of ɑ = 0.10 had a statistical power of 1 − β = 0.70. At a significance level of α = 0.15, McNemar testing would have had a statistical power of 1 − β = 0.80.Our matched case-control study of safety reports selected from VigiBase as described above, revealed an association of moderate size between pre-existing CV conditions and the reporting of ICI-related myocarditis. Drugs labeled as treatment for CV conditions, used as a proxy for concomitant CV risk factors and/or CV diseases, were associated more strongly with the reporting of ICI-related myocarditis than with the reporting of any other ICI-related ADR in VigiBase. No significant association was found between pre-existing diabetes and ICI-related myocarditis reporting. As antiobesity preparations were not mentioned in any of the included safety reports, our study did not allow an analysis of a potential association of their use with the reporting of ICI-related myocarditis. The latter two findings may be due to the relatively small size of the group of safety reports used in the study.Observational studies to date, of real-world clinical data from patient registries, pharmacovigilance databases, and claim databases, have yielded heterogeneous information on clinical risk factors for ICI-related myocarditis ,10,11,18Combination treatment is a well-established clinical risk factor for ICI-related myocarditis ,8. From With respect to ICI toxicities more in general, age subgroup analyses in randomized clinical trials (RCTs) suggested comparable safety profiles of ICIs regardless of patient age . From obIn our study, we observed that the safety reports of ICI-related myocarditis involved mostly males, likely reflecting the fact that, in RCTs, a higher proportion of men receive ICI treatment than women . The medWhile in VigiBase information on pre-existing and/or concomitant comorbidities is not systematically collected, concomitant drugs and the indications for which they are used are recorded  . Informap = 0.046) and the association between the independent variable (pre-existing CV conditions defined as concomitant drugs for the CV system— ATC C) and the dependent variable was once more found to be of moderate size  were excluded from the study group. Since patients with underlying autoimmunity may be at higher risk of ICI-related ADRs , we alsoIn accordance with previous studies, which reported about half of ICI-related myocarditis cases presenting with also other organ toxicities , we The study reported in this paper has several strengths, giving support to our conclusions. As it has a wide geographical coverage, VigiBase allowed us to involve different trends in spontaneous reporting of ICI-related myocarditis across countries. By using the MedDRA PTs of myocarditis and autoimmune myocarditis to define the dependent variable moreover, we increased study specificity . The choYet, our findings need to be interpreted in the context of some limitations. VigiBase is the global largest source of post-marketing safety reports of ADRs. Being dependent on spontaneous reporting however, safety reports from different countries and different types of reporters increase the heterogeneity of the recorded information, in terms of timeliness, completeness, and quality . In addiIn line with previous studies reporting a higher risk of ICI-related myocarditis with pre-ICI home angiotensin-converting enzyme inhibitor or angiotensin receptor blocker treatment, a history of heart failure and/or a history of acute coronary syndrome ,10, the