In this review, we aimed to tabulate the types and outcomes of the theory-based e-health interventions that had been applied to improve the e-health literacy of older adults.In this systematic review, theory-based e-health literacy interventions that published up to April 2020 were retrieved from several online electronic databases, including Medline via PubMed, Cochrane Library, ProQuest, and EMBASE. The published papers were included in this study, if the study had been conducted on older adults, a theory-based intervention aimed at promoting e-health literacy, and had been written in English language in the timeframe of 2008–2020.A total of 1658 records were identified initially, of which, 12 articles met the inclusion criteria. The systematic review identified the using of variety of intrapersonal, interpersonal, and societal level conceptual models in enhancing of e-health literacy in older adults, and the concept of self-efficacy was applied in the most of interventions as the main conceptual theoretical framework.Despite the paucity of conceptual models, which are specifically designed for e-health literacy interventions, based on our findings, we recommend self-efficacy as a powerful concept that can play an important role in improving e-health literacy in older adults. Computer-based electronic health  literacy has the potential to alleviate barriers for accessing to health care services and facilitating health care delivery . ElectroAn appropriate and effective exploration of e-health content requires some basic computer literacy and skills, including the know how to operate and maintain a computer, . HoweverConsistent with the growth of older people, an augmented demand for the improvement of e-health literacy has been reported, even in developed countries, to facilitate learning and understanding of e-health information . HoweverIn addition, during the last three decades, theory-based interventions have had significant roles in shifting research from discovering new facts and explaining events, predicting outcomes, strengthening the efficacy of findings towards successful and efficacious results; thus, behavior modification programs would be effective if and when they are developed based on a suitable theoretical framework , 15. TheThere are various theories and models that can be used to design effective interventions , which aAlthough e-health literacy interventions have benefits for older adults , 12, thehttps://www.researchgate.net/publication/343135960). Studies were searched using multiple resources to search for available relevant studies, including Medline via PubMed, Cochrane Library, ProQuest, and EMBASE.The present review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) . The proWe used the study’s research questions to guide the search terms, namely, (a) what theory-based intervention strategies have been used to improve the e-health literacy in the older adult population and (b) what outcome measures have been reported? Therefore, search terms were framed a priori, using Boolean logic. An example of the search strategy for the PubMed database was as follows:(“Computer Literacy”[Mesh]) OR (“Telemedicine”[Mesh]) OR  OR  OR  OR  OR  OR  OR  OR  OR  OR  OR  OR  OR  OR  OR (computer literacy [Title/Abstract]) OR (computer literacy [Other Term]) OR  OR  OR  OR  OR  OR online health information seeking [Other Term]) OR  OR  OR web-based health information [Title/Abstract]) OR  AND (theory [Title/Abstract]) OR (theories [Title/Abstract]) OR (model [Title/Abstract]) OR (theory-driven [Title/Abstract]) OR  OR  OR (theory [Other Term]) OR (theories [Other Term]) OR (model [Other Term]) OR (theory-driven [Other Term]) OR  OR  AND (old* adult*[Title/Abstract]) OR (old* people [Title/Abstract]) OR (elderly [Title/Abstract]) OR (aging [Title/Abstract]) OR (aged [Title/Abstract]) OR (ageing [Title/Abstract]) OR (senior [Title/Abstract]) OR (babyboomer [Title/Abstract]) OR (retiree*[Title/Abstract]) OR (pensioner [Title/Abstract]) OR (old* adult*[Other Term]) OR (old* people [Other Term]) OR (elderly [Other Term]) OR (aging [Other Term]) OR (aged [Other Term]) OR (ageing [Other Term]) OR (senior [Other Term]) OR (babyboomer [Other Term]) OR (retiree*[Other Term]) OR pensioner [Other Term]). Moreover, we examined the references of identified publications for relevant studies. This phase of the study screening yielded a total of 1169 articles after removing duplicate papers.Populations referred to studies that were included subjects 60 years and older as participants.Interventions were delimited to those in which the theoretical framework had been explicitly named, referenced, and used. The theory was defined as a set of analytical principles or statements, including defined variables, a domain to which the theory applies, and a set of relationships between the variables and specific predictions [dictions . Theory-Comparisons. We did not consider any other intervention for comparison.Outcomes were any reported impact of the theory-based intervention on the improvement of e-health literacy.Time. All articles published between January 2008 to April 2020 were considered.Setting. There was no limitation based on the type of settings.The study’s eligibility criteria were formulated, a priori, utilizing the PICO  framework, and the content validity was examined and approved by members of the research team :The eligible study designs included English language quantitative , time series, and before-after studies), or mixed methods (focusing on the quantitative strand) studies. Systematic reviews were not included but used to identify additional eligible studies. Studies that referenced a theory, model, or framework but did not discuss it and/or provided no empirical evidence to support the effectiveness of the intervention were not included in our study.A final comprehensive search strategy, in accordance with the Peer Review of Electronic Search Strategies statement , was devThe selected studies were used to extract the necessary data. Specifically, a data extraction form was developed with input from the research team to collect information on study characteristics, demographic information, aim, theoretical framework, and the use of the theory in identifying determinants, selecting/tailoring interventions, and evaluating the impact of the intervention(s). Additionally, in case of extracting of qualitative details in the review process, the findings were synthesized using a qualitative narrative approach.Two members of the research team, SP and ZF, independently pilot-tested the data extraction form, utilizing two of the nineteen articles, compared and discussed the findings, and the feedback was used to refine the form. The final draft of the form was used by SP to extract data from the remaining seventeen articles, which were independently checked by ZF. Next, both of SP and ZF reviewed the full texts of the articles and cross-validated the eligibility based on the aforementioned inclusion criteria. Subsequently, eleven of the nineteen articles was eliminated, resulting in a final sample of eight studies. Articles were eliminated if not reporting empirical data , 23, notThe qualitative assessment was conducted independently by two reviewers (SP and ZF) using the CONSORT and TREND checklist. These checklists consist of 25 and 22 criteria, respectively. If a study fulfilled a criterion it received one point. A higher overall score indicated lower methodological bias. The obtained risk of bias score of each study was divided by 25 in CONSORT and 22 in TREND (highest attainable score) and multiplied by 100 to obtain the percentage of fulfilled criteria. The disagreement between the reviewers was resolved through discussion and consensus with a third reviewer (HA). Studies were then grouped into low (> 66.7% fulfilled criteria), moderate (50–66.7% fulfilled criteria), and high risk of bias (< 50% fulfilled criteria) , 33.n = 12). Six studies used a NRSI designs [Table  designs , 42, 43. designs  to 272 p designs . The par designs  to 101 [ designs , with th designs , 41–43,  designs .Table 1n = 6 studies) had low risk of bias (range = 68–81%) [On average, the studies fulfilled 65.7% of the assessment criteria (range = 54–81%). Hence, overall the studies had a moderate risk of bias and over half of the studies ( 68–81%) , 40, 41. 68–81%) , 42, 43.Several general conceptual frameworks were used across the studies to improve e-health literacy in older adults. Specifically, the Health Belief Model (HBM) in health behavior research , the selSeveral intervention tools and/or manuals were used to promote e-health literacy of older adults. Some studies used instructional manual developed by the National Institute on Aging (NIA) of the National Institutes of Health (NIH) , 41, 43.Interventions were intrapersonal, interpersonal, or societal, focusing on improving the participants’ e-health literacy via the instructor/teacher-center and/or interactive/collaborative/peer tutor delivery approaches. Five studies used didactic sessions and workshops in classroom settings , 42, 43,http://nihseniorhealth.gov/toolkit/toolkit.html), which was used in the studies conducted by Xie, had a number of features to improve the individual’s self-efficacy in using a computer to access e-health. In Xie’s toolkit, each training session was created based on the information from the previous session and gradually added to the complexity of lessons and goals. Therefore, the knowledge and skills of learners increased gradually during the sessions, which assisted the participants in competency building and computer self-efficacy [Self-efficacy model was the common conceptual model used nearly in all the e-health literacy improvement interventions. For example, the NIA toolkit  in partnering with seniors for better health program, used instructors to develop older adults’ computer literacy and increase their skills in retrieving and appraising online health information. By breaking the skills to simpler steps and gradually adding more complex steps, instructors not only enhanced the participants’ self-efficacy, but also enabled them to perform a more complex Internet search for accessing e-health information .Fink and Beck used the constructs of HBM to develop e-Health literacy by improving the online searches abilities. For example, they increased the users’ perceived threat by demonstrating the pitfalls of relying on public search engines. To increase the participants’ self-efficacy, they provided user-friendly guidelines for evaluating websites quality, provided a list of high-quality websites, and illustrated the benefits of using high-quality websites .Cooper-Gaiter (2015) organized a facilitator-led computer workshop and training modules at a local community center. After the workshop, she obtained favorable results in increasing participants’ self-efficacy in using e-health information .In addition, Nahm et al. (2019) emphasized on four sources of efficacy beliefs, namely, (1) successful performance of the behavior, (2) social persuasion, (3) modeling others’ successful performances, and (4) relief of emotional stress , which wBased on another approach, Bo Xie (2011) used the social interdependence theory as a conceptual framework to improve the e-health literacy of the older adults by developing collaborative learning tactics –36. CollFinally, Chiu et al. (2016) used TAM and DOI to enhance the perceived usefulness of internet technology in accessing e-health. To increase older adults’ acceptability of e-health and reduce their perceived barriers in utilizing complicated computer application, they selected and downloaded several simple e-health computer applications. In addition, they provided a step-by-step training illustrations which indicating the applying of self-efficacy development in this program .Several studies showed an improvement in the older adults’ level of interest, confidence, and self-efficacy in using e-health information , 40–43. We also reviewed the conceptual models used in designing e-health literacy interventions in older adults. Among the existing health behavior models, self-efficacy was the most frequently cited conceptual model. However, none of these studies used the conceptual frameworks that were specifically designed for e-health literacy intervention, such as the Lily model or the framework proposed by Chan and Kaufman , 45. ThiSelf-efficacy refers to a personal belief in his or her capacity to perform behavior/behaviors that is/are necessary to attain specific performance . AccordiWatson believed that one of the factors that might affect to use less the online health information by older adults, is the lack of internet self-efficacy . InterneIn addition, it is postulated that personal self-efficacy beliefs can be improved via one’s sources of information regarding the establishment, reinforcing or weakening of such beliefs, potentially influencing individual’s behavior . BanduraThe main source of self-efficacy is mastery experience . TherefoThis systematic review’s delimitations and limitations must be acknowledged. The sample was delimited to articles published in English, so it may not reflect findings of studies that were published in other languages. The keyword search was restricted to the title, keywords, and abstract for each article. Due to the high variability in the intervention tools, strategies, and outcome, it was not possible to perform a joint statistical analysis of the data. Consequently, a narrative analysis was conducted, which limited the external validity of the findings.The aforementioned findings suggest that the application of the conceptual frameworks in the context of e-health literacy interventions has the potential to enhance the outcome measures. Meanwhile, they suggest a need to empirically test the efficacy of the conceptual models that are designed to specifically improve e-health literacy. Based on our findings, we recommend self-efficacy as a powerful concept that can play an important role in improving health literacy in older adults by increasing their confidence in themselves and their abilities to use the Internet and web-based learning despite their age-related limitations.


MYCN amplification (MNA) or dysregulation. MYCN is associated with poor prognosis and its over-expression leads to several dysregulations including metabolic reprogramming, mitochondria alteration, and cancer stem cell phenotype. Some hints suggest that MYCN overexpression leads to cancer immune-escape. However, this relationship presents various open questions. Our work investigated in details the relationship of MYCN with the immune system, finding a correlated immune-suppressive phenotype in neuroblastoma (NB) and different cancers where MYCN is up-regulated. We found a downregulated Th1-lymphocytes/M1-Macrophages axis and upregulated Th2-lymphocytes/M2-macrophages in MNA NB patients. Moreover, we unveiled a complex immune network orchestrated by N-Myc and we identified 16 genes modules associated to MNA NB. We also identified a MYCN-associated immune signature that has a prognostic value in NB and recapitulates clinical features. Our signature also discriminates patients with poor survival in non-MNA NB patients where MYCN expression is not discriminative. Finally, we showed that targeted inhibition of MYCN by BGA002 (anti-MYCN antigene PNA) is able to restore NK sensibility in MYCN-expressing NB cells. Overall, our study unveils a MYCN-driven immune network in NB and shows a therapeutic option to restore sensibility to immune cells.A wide range of malignancies presents  MYCN is a transcription factor member of the MYC proto-oncogene family involved in nervous system development during embryogenesis  is established as a major driver of Neuroblastoma (NB) (characterizing 50% of the high-risk group) (k group) –13. NB ik group) . Currentk group) –17.Immune evasion plays a fundamental role in the development and progression of cancers and is driven by the tumor microenvironment remodeling by cancer cells . DiffereHowever, while anti-GD2 therapy and chimeric antigen receptor (CAR) T cells showed some promising results in non-MNA NB patients, the checkpoint inhibitors have not shown the same success in improving the survival in NB, as in other solid tumors. These poor results can be linked to different factors as low MHC-I expression, low presence of neoantigens, immunosuppressive environment , 26–28. MYCN impact on the immune system. Indeed, the NB immunity and the role of MYCN in the immunosuppression are still a field of investigation  and processed using the quantile algorithm in limma. Another NB dataset and Wilms’ dataset were downloaded from TARGET data portal . From NCBI GEO DataSets (https://www.ncbi.nlm.nih.gov/gds) were downloaded the following datasets: retinoblastoma (accession: GSE59983), rhabdomyosarcoma (accession: GSE114621), T acute lymphoblastic leukemia (T-ALL), and acute myeloid leukemia (AML) dataset  and for the T-helper lymphocyte (Th) profiles (accession: GSE107011). Another SCLC dataset was retrieved from the supplementary of the article (PMID: 26168399)  patients and non-MNA patients in the NB datasets  were obtained using the limma package algorithm .The NB dataset (accession: E-MTAB-1781) and the small cell lung cancer (SCLC) (accession: E-MTAB-1999) datasets were downloaded from ArrayExpress, (6168399) . In the MYCN or MYC in each dataset were used as ranked gene list to identify enriched pathway through Gene Set Enrichment Analysis (GSEA) (Correlated genes with s (GSEA) . We usedWe used CIBERSOFT tool (Cell type Identification By Estimating Relative Subsets Of known RNA Transcripts) as described in the developer instruction . NB exprMYCN knock out genes were downloaded by KnockTF  from E-MTAB-1781 were filtered for this list. WGCNA  was perfversus not MNA patients. The model was cross-validated 50-fold using E-MTAB-1781 as training set (80% of observation at each run) and using TARGET dataset as test set. We used L1 penalization (C = 0.1) and SAGE solver. We selected the weights for each gene and averaged . We selected two different vector weights associated with MYCN, positive weight vector (associated with MNA) and negative weight vector (associated with non-MNA). The two vectors were normalized subtracting the minimum and dividing by the range:Immune genes retrieved from Gene Ontology (GO) and literature. Patient gene expression profiles (GEP) from E-MTAB-1781 and TARGET were filtered for this list (gene list is present in Through univariate cox regression we selected genes significantly associated to the prognosis (we used the same gene list used for the logistic regression model). The obtained p-value was correct with the Bonferroni correction (list of significant genes obtained through univariate cox regression is present in MYCN and Cox model. We then build a positive and a negative immune score for each dataset (-MTAB-1781 and TARGET) multiplying each gene x (log2 expression) for each normalized weight  and MYCN abstracts querying PubMed . Clinical information was retrieved within the dataset  and Python (Anaconda release). The following libraries from Python (version 3.7) were used: Scikit-learn, Matplotlib, matplotlib.pyplot, Pandas, UMAP, numpy. The following libraries from R (version 3.5) were used for analysis and graphs: ggplot2, dplyr, data.table, tydr, survival, survminer, wordcloud, WGCNA, circlize, iGraph, anRichment, stringr.The cell lines used in this study were obtained in 2020 and kept in culture for 30 days and seven passages at maximum. Mycoplasma detection was conducted with LookOut Mycoplasma PCR Detection Kit (Sigma-Aldrich). Additional details about the cell line used in this study can be found in Kelly-luc cell-line (Kelly NB cell line transfected with luciferase gene) was generated as described in . Kelly-lMYCN in NB, we performed GSEA analysis in NB patient datasets. Interestingly, MYCN negative correlated genes are significantly enriched of different immune system pathways in both NB cohort 1 (E-MTAB-1781) and NB cohort 2 (TARGET)  in MNA NB. Since, MYCN overexpression is present in a large group of tumors . We observed that different pathways associated to Th1 are negatively correlated to MYCN in different cancer types . Remarkably, the high MYCN immune dysregulation group was associated with poor prognosis while the low group with a favorable prognosis in NB . Anti-MYCN BGA002 potently reduced MYCN expression and led to a significant CD276 down-regulation after the treatment in MYCN-expressing MNA and non-MNA NB cell lines  expression has been reported in NB, but PD-L1 blockade immunotherapy has not reported to be effective in NB . We did MYCN immune signature and highly expressed in MNA NB patients . We did not notice viability decrease adding the NK-cells alone in co-culture with MNA-NB cells   . Interesr growth . Indeed,MYCN expression regulated different genes involved in direct interactions with immune cell types playing as a driver of this poor immune environment. We identified 16 immune gene modules that are differently enriched in MNA and non-MNA NB patients, where modules related to immune receptors, signaling, and cytokines are enriched in the latter group. Therefore, all these results confirm a deeply dysregulated immune tumor micro-environment in NB. We also inferred the putative TFs that regulate the immune genes and differed between MNA and non-MNA NB, founding three regulon clusters  who are in common between NB1 and NB2 cohorts. Our results show that these regulon clusters are differentially enriched in MNA and non-MNA patients and they regulate the immune landscape in NB. We found that there are direct interactions between these TFs, as well as N-Myc also directly regulates the expression of these regulons as mechanistic explanation of the immune response dysregulation in NB.We showed that mapping the possible protein-protein interactions between immune cell types in non-MNA NB patients revealed a complex immune network that is lost in MNA patients. Moreover, we found that MYCN immune dysregulation and prognostic impact in NB. Generating a MYCN immune signature we stratified the NB cohorts in three groups  which showed a marked difference in the prognosis. Moreover, the MYCN immune score was also associated with other different NB characteristics as stage, proliferation, and histology and we confirmed these associations in an independent NB cohort. As a confirmation, the MYCN immune score correlated with immune checkpoints, Th cytokines, MHC genes, and TLRs capturing the immune landscape of the NB. Moreover, apart from MNA, there are other cancer events that can lead to dysregulated N-Myc higher activity  making difficult to infer the MYCN relevance in these cases , showing MYCN inhibition and a therapeutic effect in vitro and in vivo (MYCN. Our results showed that MYCN inhibition by BGA002 resulted in a cascade to downregulation of negative immune checkpoints (CD276) and regulons implied (HMGA1) in the immune-suppression phenotype. Indeed, we noticed that anti-MYCN treatment also led to NK lysis of MNA NB cells.MNA and refractory NB patients are lacking viable therapeutic options , 68. Sinrvention . Despite in vivo , 10. ThuMYCN in suppressing the immune landscape, which play a role in the poor prognosis associated to this oncogene. These data also suggest that MYCN blocking can ameliorate the immune suppression characterizing MNA NB patients. Indeed, while specific MYCN inhibition by anti-MYCN BGA002 can be proposed as a single treatment for MNA NB patients, our results also show that its activity can restore the responsiveness of the immune system against NB, opening the way to use anti-MYCN inhibition in combination with immune-therapy.Collectively, the data here presented provide demonstrations of the broad role of The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/The studies involving human participants were reviewed and approved by the Scientific Ethical Committee of Bologna University. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.SR performed the bioinformatic analysis. CA, SL, and LM performed the experiments and acquired and analyzed the biological data. MF provided dataset for the analysis. SR, DR, and RT wrote the manuscript. AP, PH, and MF revised the manuscript. AP, PH, MF, and RT acquired funds. SR and RT designed and supervised the study. All authors contributed to the article and approved the submitted version.RT, AP, and PH are funded by the University of Bologna. LM is funded by AGEOP. SR, CA, and SL are funded by BIOGENERA SpA; the funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.RT and AP are co-founders and shareholders of Biogenera. Authors SR, SL and CA are employed by Biogenera.The remaining 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.


Despite barrier measures and physical distancing tailored by the populations worldwide, coronavirus continues to spread causing severe health and social-economic problems. Therefore, researchers are focusing on developing efficient detection and therapeutic platforms for SARS-CoV2. In this context, various biotechnologies, based on novel molecules targeting the virus with high specificity and affinity, have been described. In parallel, new approaches exploring nanotechnology have been proposed for enhancing treatments and diagnosis. We discuss in the first part of this review paper, the different biosensing and rapid tests based on antibodies, nucleic acids and peptide probes described since the beginning of the pandemic. Furthermore, given their numerous advantages, the contribution of nanotechnologies is also highlighted. To date, six human coronaviruses have been identified: α-coronaviruses , β-coronaviruses , severe acute respiratory syndrome-CoV (SARS-CoV), and Middle East respiratory syndrome-CoV (MERS-CoV) [Different measures have been imposed by authorities, including wearing masks and physical distancing, aiming to stop the virus spread. However, the adopted procedures are not sufficient, and mass screening of coronavirus is more than a necessity. At present, real-time RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction) is the most used technique for COVID-19 testing. It is based on specific primers targeting sequences encoding the proteins E, N, S, ORF1b(Open Reading Frame 1b), and RdRp(RNA-dependant RNA polymerase)or a combination of different targets ,5. The mApart from familiar technologies, there have been several reports describing the identification of new biorecognition elements that have been employed in different biosensing schemes ,11. In aSeveral research groups are working on the development of rapid and sensitive assays and biosensors to overcome the limitations of traditional techniques used in coronaviruses detection. These techniques are mainly based on antibodies and nucleic acids including aptamers, clustered regularly interspaced short palindromic repeats (CRISPR), and antisense oligonucleotides.Currently, immunoassays are the most popular diagnostic tools available in the market and used in medical structures. Basically, these methods use antibodies as bioreceptors targeting capsid proteins or whole viruses. In serological testing, capsid proteins are used as viral antigens to bind the immunoglobulins generated by the patient against the pathogen. Antibodies are usually obtained from animal immunization with N, S, or E protein or from the blood samples of patients who are infected . In addiElectrochemical immunosensing is based on the conversion of the antibody–antigen interaction into electrochemical signals. In contrast to traditional immunoassays, innovative transducers allow a highly sensitive quantification of the targeted antigen/antibody in different ways . It shouField-effect transistor(FET)-based biosensing is characterized by many advantages, including sensitivity, fast response, and low-noise detection . MoreoveBesides electrochemical methods, optical biosensors have been widely used in clinical diagnosis and drug discovery due to their analytical performance in detecting biological systems . DiffereSurface plasmon resonance-based biosensors are based on the monitoring of refractive index variations after the formation of the target–receptor complex on the sensing surface. Plasmonic biosensing offers many advantages, including label-free and real-time monitoring . Yong’s Besides label-free sensors, optical sensors, based on chemiluminescent and colorimetric labeling, have been reported for COVID-19 diagnosis. First, Cai et al. developed a chemiluminescent enzymatic serological immunoassay based on a peptide from the S protein. The latter was screened from peptide antigens synthesized from the genomic sequence from GenBank (NC_045512.1). Streptavidin-functionalized magnetic beads were used as an immobilization support for the biotinylated peptide. The binding between IgM and IgG present in serum samples and the magnetic beads was carried out in solution. After binding and washing, the conjugate was reacted with the substrate, generating a luminescent signal, which was measured by a luminometer. In this work, no clinical application was carried out . AnotherDespite the importance of IgGs in detecting previous COVID-19 infection and the association of IgM to recent contamination, the role of secretory immunoglobulins IgA should not be neglected. IgAs(Immunoglobulin A)play a key role in the immune exclusion process limiting the access of microorganisms to the mucosal barriers. It has been demonstrated that elevated total IgA and IgA-aPL(IgA-antiphospholipid) are significantly associated with severe COVID-19 infection ,33. RodaThe different immunosensing strategies described above are summarized in As coronaviruses are positive single-stranded RNA viruses, their detection is usually based on nucleic acid techniques, mainly real-time PCR. However, this technology requires expensive reagents and equipment, in addition to qualified manipulators. Therefore, researchers are focusing on new techniques based on clustered regularly interspaced short palindromic repeats (CRISPR), aptamers, and nucleic acidhybridization to reduce the dependence on RT-PCR.After the identification of CRISPR-Cas systems targeting RNA, numerous studies have shown the potential use of this tool in the detection of different viruses such as dengue and Zika viruses . Cas13 eThe first designed and most used platform, called SHERLOCK showed a successful application for identifying cell-free tumor DNA mutations, genotyping human DNA, detectingdengue and Zika viruses, and distinguishing pathogenic bacteria . SeveralBy combining SHERLOCK with the HUDSON protocol; a process for lysis of viral particles and inactivation of DNases and RNases in body fluids with heat and chemical treatment, no prior RNA extraction is needed . In a reApart from Cas-13, Cas-12 effectors have been also employed for coronavirus diagnosis. For that, CRISPR-Cas12 was combined to DETCTR platform with the DNA endonuclease-targeted CRISPR trans reporter (DETCTR) platform. Extracted RNA is first retro-transcripted to DNA, followed by isothermal amplification using the RT-RPA enzyme. The Cas12a-gRNA complex is then activated through identification of a specific sequence in the amplified DNA, leading to the cleavage of reporters as it was described for the SHERLOCK platform. Based on this principle, different fluorescent lateral flow assays have been developed and tested in clinical samples, showing excellent results .CRISPR-Cas9 effectors have been also exploited to construct a triple line lateral flow assay for the simultaneous dual gene (E and Orf1ab) detection of SARS-Cov2. The assay is based on the recognition between the Cas9/sgRNA and DNA-AuNPs probes. After binding, the formed complex accumulates on the test line and generates a visible colorimetric signal. The originality of this strip is the simultaneous detection of more than one gene which is not available by using Cas13 and 12 requiring nonspecific cleavage .Finally, these studies confirm that CRISPR technology holds a great potential for the diagnosis of the novel coronavirus. It constitutes a promising alternative for the RT-qPCR technique, mainly due to its low cost, simple fabrication and sensitivity.Aptamers are a class of bio-inspired receptors composed of single stranded DNA or RNA. They are selected by a combinatorial process called selection of ligands by exponantial enrichment (SELEX)for their affinity and specificity to a target . They arIn April 2020, Song et al selected the first aptamer specific for receptor binding domain of the SARS-CoV-2 spike glycoprotein. SELEX was carried out by incubating a ssDNA (single stranded DNA) library comprising 40 random nucleotides with RBD protein immobilized on A-beads. After a total of twelve rounds of selection and amplification, two sequences have been chosen. They exhibited a high affinity for the novel coronavirus with respective Kd values of 5.8 and 19.9 nM. In addition to the experimental competition, molecular dynamic simulations suggested that the selected aptamers bind to identical amino acids of RBD, thus providing a promising issue for coronavirus diagnosis, prevention and treatment . The feaApart from the spike protein, Nucleocapsid protein (NP) has been also targeted by SELEX technology. Zhang et al selected four aptamers specific for N protein with high affinity lower than 5 nM.For that, a total of five rounds of selection were performed starting from a library of 76 nucleotides comprising a randomized sequence of 36 nucleotides. After being selected, aptamers were employed in a sandwich-type assay based on four aptamer pairs, which form a ternary complex with NP. Combined with antibodies, this principle was first applied in an enzyme linked immunosorbent assay showing a high sensitivity. Then, an immunochromatographic strip was developed using gold nanoparticles to allow the visual detection of NP in urine and serum . The selBesides to aptamers specifically selected for the novel coronavirus, the potential use of SARS-CoV-1 specific aptamers has been also investigated. Based on the fact that the sequence of SARS-CoV-2 N protein exhibits a homology of 91% with that of SARS-CoV, Chen et al studied the binding capacities of the SARS-CoV aptamer to detect the new coronavirus. Enzyme-linked aptamer binding assay (ELAA) demonstrated that the three tested aptamers exhibit a binding affinity for the SARS-CoV-2 N protein opening the way for their application in COVID-19 diagnosis and treatment. Aiming to validate that, the binding efficiency of aptamer 2 was compared to that of a commercial antibody using Western blot showing similar results. These aptamers could thus be a good alternative for antibodies and immunoassays, currently used for coronavirus detection. Moreover, the use of these aptamers should provide a low cost and rapid solution by avoiding the selection of new candidates specific to SARS-CoV-2 .Recently, an interesting study was published by exploring the binding affinity of a nucleocapsid protein specific aptamer and proximity ligation strategy. The principle is based on targeting the antigenic protein with two different aptamers in order to bring the ligation DNA region into close proximity and initiate ligation-dependent qPCR(quantitative PCR) amplification . The proDespite the excellent characteristics of aptamers, few research reports explored their advantages in COVID-19 diagnosis. Based on the excellent results of the aptasensors discussed above, more techniques will be certainly described in the future by exploring the different sequences selected either for SARS-CoV or SARS-CoV-2.Based on the predictable and specific hybridization of complementary bases, DNA-RNA hybridization has been widely used in RT-PCR and other biomedical diagnosis techniques. Hybridization is mainly dependent on melting temperature; complementary strands can hybridize with each other when the temperature is slightly lower than their melting temperature, while a single mismatch can cause the melting temperature to decrease significantly . Qui et Owing to their simple synthesis and low cost, antisense oligonucleotides could be more advantageous than the other bioreceptors discussed in this review including aptamers and antibodies.In recent years, nanotechnology and nanoengineering have shown a remarkable progress. Nanomaterials have been widely studied and applied in numerous fields including medicine, showing great potential in the diagnosis, prevention, control and treatment of many diseases ,85. ThisAfter discussing the different COVID-19 biosensing platforms integrating nanomaterials in the first part of this review, we focus in this part on the different therapeutic strategies based on nanotechnology. Basically, nanoparticles can be used as drug carriers and viral inhibitors. Moreover, they hold a great potential in vaccine research for the novel coronavirus.Nanomaterials have been reported as excellent carriers for delivering drugs, with high selectivity, to targeted locations in organism. It has been demonstrated that nanocarriers reduce therapeutic agent toxicity, immunogenicity and side effects while increasing drug efficacy. Moreover, certain nanomaterials have been already approved by the FDA for drug delivery such as polylactic-co-glycolic-acid and lipid nanoparticle systems . In vaccine design, nanocarriers constitute a key element either in antigen encapsulation and presentation or boosting of the immune system via the co-delivery of antigen and adjuvant . The abiBesides to vaccine delivery, nanomaterials could be used as antiviral carriers for SARS-CoV-2 treatment. For instance, Ullah et al. proposed a siRNA-based antiviral strategy by using cationic liposomes as a delivery system. Aiming to target ACE2 receptors, the authors proposed an aerosol formulation for intranasal or intratracheal administration. This targeting can be achieved by functionalizing lipid/polymer nanocarriers with highly specific receptors such as antibodies or aptamers. Such a strategy could promote the use of siRNA for silencing and knockdown of viral genes .2, SiO2…) have been used as antivirals against different viruses including hepatitis B, H3N1 and H1N1viruses [Nanomaterials can be successfully conjugated to various functional groups, linkers and bioactive molecules. Moreover, it has been demonstrated that nanoparticles are able to cross the blood-brain and blood-air barriers as well as lymphatic system ,95. Furt1viruses . For ins1viruses .Based on these findings, nanomaterials-based antiviral strategy could be used as promising tool for SARS-CoV-2 inhibition. In the literature, different reviews have discussed the potential of nanotechnology in combating COVID-19 infection ,99. In tThe health crisis caused by the COVID-19 pandemic showed the key role of biotechnologies in biomedical research. As COVID-19 patients display nonspecific symptoms, efficient diagnostic approaches are highly required. Many efforts have been devoted worldwide to meet the urgent need for cost-effective mass screening. In this context, a large array of biosensors has been developed based on different bioreceptors and detection techniques. Many research groups have investigated immunochemical techniques based on antibodies, peptides and aptamers, combined with electrochemical and optical transducers for the sensitive and specific detection of viral antigens and/or immunoglobulins. In parallel, viral genes have also been targeted by using different types of nucleic acids including CRISPR and antisense oligonucleotides. Most part of the developed biosensors was portable allowing rapid, cheap and easy to use point of care testing of SARS-CoV-2. Moreover, some techniques allow the sensitive determination of more than one indicator providing a complete diagnosis and monitoring platform. Furthermore, dual platforms combining simultaneous sample collection and antigen sensing have also been reported. Finally, the selectivity of these technologies has been demonstrated against different types of human pathogens. Besides diagnosis, vaccination is also of a great importance in preventing the propagation of viruses, in particular, with the fast spreading UK virus variant which is raising alarms . In this


It is well recognized that cell-type heterogeneity hampers the interpretation of Epigenome-Wide Association Studies (EWAS). Many tools have emerged to address this issue, including several R/Bioconductor packages that infer cell-type composition. Here we present a web application for cell-type deconvolution, which offers the functionality of our EpiDISH Bioconductor/R package in a user-friendly GUI environment. Users can upload their data to infer cell-type composition and differentially methylated cytosines in individual cell-types for a range of different tissues.https://www.biosino.org/EpiDISH/.EpiDISH web server is implemented with Shiny in R, and is freely available at  Epigenome-wide association studies (EWAS) aim to identify epigenetic alterations associated with disease, or with environmental risk factors. Most EWAS have been conducted in complex tissues  that are made up of many different cell-types. Since DNA methylation (DNAm) is highly cell-type specific, cell-type heterogeneity can lead to confounding . PreviouThe web server is depicted in Users are required to upload their DNAm beta value matrix, phenotype of interest (POI) file  and covariates matrix . Both .txt and .csv formatted files will be accepted. After uploading, a preview will automatically appear on the right side of the page, so users can check that everything is ok. A prompt message will tell the users about the dimensions of the uploaded data.We provide references for three tissues types, including whole blood, general epithelial tissues and breast tissue. For each of them the user can select the cell-types for which they would like to estimate fractions. For general epithelial and breast tissue, users can specify whether they would like to know the fractions of immune cell subtypes, as we otherwise return the total immune cell fraction. Users can also choose the estimation method from: robust partial correlation . Users cP-value threshold to call DMCTs  and the NSFC ; L.M., W.Y. and G.Z. are supported by National Key R&D Program of China .Conflict of Interest: none declared.


The issue contains material on classical sequence analysis, bioinformatics applications in medicine, and the theoretical research on gene network structure and dynamics. This traditional school on bioinformatics is organized annually since 2008 by the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences and Novosibirsk State University. In 2019 we had several large international meetings in Russia - in Novosibirsk and Moscow presenting materials on genetics and bioinformatics. The name of the special issue - SBB-2019 reflects the main topics of the event. Traditionally we select the best conference and school materials to be presented at BMC Genomics  and BMC Genetics post-conference publications [https://bmcbioinformatics.biomedcentral.com/articles/supplements/volume-20-supplement-1).This Special Issue of BMC Bioinformatics “Systems Biology and Bioinformatics” collates the papers presented at the 11th Young Scientists School “Systems Biology and Bioinformatics”-2019 (SBB-2019) hold in summer 2019 in Novosibirsk, Russia  [https://bgrssb.icgbio.ru/2020/). For the first time, it was in an online format. We believe such events and public discussion at the platforms of international publishers bring attention of the readers [https://www.frontiersin.org/research-topics/14266/bioinformatics-of-genome-regulation; https://peerj.com/collections/72-bgrs-sb-2020/).The SBB Schools in Novosibirsk are satellite events for BGRS\SB multiconference organized biannually. In 2019, the SBB-2019 School was held as a broad-scope independent meeting with science and education components. Other Special Issues in the fields of genomics, bioinformatics, microbiology, and medical genomics accompany this Special Issue in bioinformatics, published as a part of the following series: BMC Genomics and BMC Medical Genomics, BMC Genetics and BMC Medical Genetics, as well as in BMC Microbiology. In 2018, the conference highlights were organized into the Special Issues with reports from the BGRS\SB-2018 conference and earlier from Belyaev Readings-2017 (v100/en) , 6. We cThe papers comprising this issue of BMC Bioinformatics were discussed at the SBB-2019 School in Novosibirsk. We open up this Special Issue by classical work on protein sequence analysis by Valery Polyanovsky and colleagues  . The original work on the dynamic of cell replications [Tamara Khlebodarova and colleagues applied gene network models for the analysis of dynamic regulation of synaptic protein that may cause neuropsychiatric diseases  . Next, SBB-2020 School in Russia is scheduled for September 2020. BGRS\SB-2020 (Bioinformatics of Genome Regulation and Structure \ Systems Biology) multiconference just over in July 2020 holding series of symposia and workshops (https://bgrssb.icgbio.ru/2020/), and following journal publications . We invite our readers worldwide to attend our next events on Systems Biology and Bioinformatics in Russia, as well as the systems biology meetings in Moscow - Digital Medicine Forum .Therefore, this issue includes reports of recent bioinformatics applications in protein sequence and structure analysis, text mining, gene networks modeling. We aim to support international exchange and education in new forms via the schools and competitions for young scientists (


Subgroup analysis showed that the C-reactive protein-to-albumin ratio is a significant prognostic marker for various head and neck cancers. An elevated pretreatment C-reactive protein-to-albumin ratio predicts a worse prognosis for patients with head and neck cancers. Therefore, the C-reactive protein-to-albumin ratio could serve as a potential prognostic biomarker facilitating treatment stratification.The C-reactive protein-to-albumin ratio is a proven prognostic predictor of nasopharyngeal carcinoma. However, the role of the C-reactive protein-to-albumin ratio in other head and neck cancers remains unclear. This meta-analysis explored the prognostic value of the C-reactive protein-to-albumin ratio in head and neck cancers. A systematic search was conducted. Outcomes of interest included overall survival, disease-free survival, and distant metastasis–free survival. The hazard ratio with 95% confidence interval was pooled using a random-effects model. A total of 11 publications from the literature were included, allowing for the analysis of 7080 participants. Data pooling demonstrated that pretreatment C-reactive protein-to-albumin ratio had a hazard ratio of 1.88 (95% CI: 1.49−2.37,  Head and neck cancer (HNC), including cancers of the nasopharynx, oral cavity, oropharynx, hypopharynx, and larynx, accounts for 800,000 of all new cancer diagnoses each year and caused more than 400,000 deaths in 2018 ,4.Accumulated evidence has demonstrated the roles of systemic inflammation and host immunity in angiogenesis and cancer progression ,6. StudiTwo meta-analyses have reported the prognostic value of CAR for determining nasopharyngeal cancer (NPC) survival ,15. HoweA systemic review and meta-analysis was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis criteria . A strucPublished research that satisfied the following inclusion criteria was included: (1) it explored the relationship between pretreatment CAR and long-term prognosis, including overall survival (OS), disease-free survival (DFS), or distant metastasis–free survival (DMFS) in HNC; (2) patients did not undergo any oncologic management such as operation or neoadjuvant therapy before CAR samples were obtained; and (3) the article reported necessary information for a meta-analysis. We excluded (1) studies that did not include survival outcomes and (2) letters, epidemiological studies, case reports, review articles, conference abstracts, and duplicate publications. Two authors (Chih-Wei Luan and Hsin-Yi Yang) independently reviewed the citations among the identified studies and enrolled studies that satisfied our inclusion criteria. The references were checked to identify potential eligible studies. A third author (Yao-Te Tsai) adjudicated on disagreements between the first two authors.The authors (Chih-Wei Luan and Hsin-Yi Yang) independently reviewed the included studies and extracted the following data using a standardized data collection form: study details (author(s), sample size, sex, publication year, study design (retrospective or prospective), and country of study), pathological characteristics (TNM staging), and clinical features . The CRP/Alb ratio was calculated by dividing the serum CRP level by the serum albumin level with same scales as follows: CRP level (expressed in mg/L)/albumin level (expressed in g/L). A third author (Yao-Te Tsai) arbitrated when the first two authors disagreed.The authors (Chih-Wei Luan and Hsin-Yi Yang) independently evaluated the quality of the included studies using the Newcastle–Ottawa quality assessment scale . Studies2 statistic. All data pooling was performed using a random-effects model. Metaregression and cumulative meta-analysis by publication year, sample size, and cutoff values were employed to examine the potential influence of these factors on effect estimates. Subgroup analysis was performed to examine the significance of various potential moderators, including tumor sites, sample size, country of publication, cutoff values, and follow-up periods. To assess publication bias, Begg’s, Egger’s tests and a funnel plot were conducted. A p value of < 0.05 was defined as statistically significant in all calculations. Statistical analyses were conducted using STATA software version 15 .The primary outcome measure of this study was the OS of patients with HNC. Secondary outcomes were DFS and DMFS. We pooled hazard ratios (HRs) with their corresponding 95% CIs to estimate the association between CAR and survival outcomes and determined heterogeneity using Cochran’s Q-test and the IA structured search returned 110 records from PubMed, Embase, and CENTRAL. The flow diagram in p < 0.0001) for predicting mortality. The results suggest that a higher pretreatment CAR significantly predicts poor OS for patients with HNC  for predicting disease recurrence or mortality compared with CARs lower than the cutoff  provided HRs for the DFS of patients with HNC based on their CAR. Regarding DFS outcomes, the pooled results revealed that a CAR higher than the cutoff had an HR of 1.87  of having a poor DMFS rate. This result implied that patients with HNC and a higher CAR might have a higher distant metastasis rate than those with a low CAR  and Egger’s tests (p < 0.001). Thus, we used the trim and fill method to evaluate the influence of publication bias on the pooled results. The results from the trim and fill calculation showed that the pooled HR was 1.57  for OS prediction after the missing studies were added. The new HR was smaller than that in the original results but did not change the direction and significance of these results. This finding supports the strength and stability of our meta-analysis.The funnel plot had apparent asymmetry on visual inspection, which demonstrates a lack of studies with small effect size HRs and small sample sizes for all three outcomes . When OSDespite previous meta-analyses providing evidence to support the prognostic value of CAR in relation to nasopharyngeal carcinoma, this is the first meta-analysis to demonstrate that CAR also has prognostic value with respect to other HNCs. Our results demonstrate that a higher pretreatment CAR is significantly associated with poor OS, DFS, and DMFS. The CAR holds prognostic prediction value for determining OS and DFS outcomes for all HNC tumor locations, including the mouth, nasopharynx, hypopharynx, and larynx . Thus, CThe sample size of this study was a significant moderator in metaregression and subgroup analysis. These analyses showed that studies with small sample sizes tend to have higher HRs. This phenomenon can be explained by the presence of publication bias. Our funnel plot showed an obvious absence of studies with small sample size and small effect estimates. This finding suggests that publication bias might exist for a study with small size and that negative findings are less likely to be published, thus leading to higher HR values in studies with small sample sizes, as identified in metaregression and cumulative analysis.In this work, tumor location and countries of origin were significant moderators in the subgroup meta-analysis. Given that only 11 studies were included in this meta-analysis, it was not appropriate to perform a multivariate metaregression to adjust for the potentially confounding influence of sample size on the moderating effect of tumor location and country of origin. However, oral cancer studies generally had smaller sample sizes than NPC studies . FurtherThe underlying mechanism of the correlation between CAR and oncologic prognosis remains uncertain. Studies have indicated that inflammation participates in tumor development and progression by affecting the microenvironment of cancer pathogenesis. Systemic inflammatory responses may influence cancer cell proliferation, apoptosis, and angiogenesis, facilitating cell invasion and exacerbating cancer metastasis through the release of cytokines, immune cells, chemokines, acute phase proteins, and small inflammatory proteins ,30,31. SThe TNM staging system, which is based on tumor factors, is always a crucial reference for treatment planning and prognostic prediction. However, in that system, host factors are not considered, such as nutritional status and systemic inflammation, which may account for disparate prognoses in patients with the same cancer stage. Therefore, despite similar TNM staging and advances in HNC therapy, equivalent treatment approaches may fail and lead to locoregional recurrent and distant metastasis.Developing individualized treatment modalities with appropriate regimens is still difficult and often controversial. By assessing patients’ CAR, physicians can choose tailored treatment regimens for patients with HNC and inform their patients about management and long-term results ,37. ReceThis meta-analysis had several limitations. First, included studies had considerable methodological diversity. Of the 11 included studies, tumor site, cancer stage, treatment method, CAR cutoff values, and follow-up periods all differed. Such diversity might have caused the observed statistical heterogeneity. Second, all included studies had exclusively Asian patient populations. Thus, it is difficult to generalize our findings to the global population. Third, the great variation in the cutoff value of CAR hinders its clinical application. Thus, future studies are needed to identify the exact cutoff of CAR to be clinically useful. Finally, this meta-analysis had obvious publication bias due to the lack of small studies with negative findings. However, our trim and fill analysis showed that publication bias did not alter the results of our meta-analysis. Nonetheless, we encourage the publication of small studies with negative findings.


With the continued spread of COVID-19 across the world, rapid diagnostic tools, readily available respurposable drugs, and prompt containment measures to control the SARS-CoV-2 infection are of paramount importance. Examples of recent advances in diagnostic tests are CRISPR technology, IgG assay, spike protein detection, and use of artificial intelligence. The gold standard reverse transcription polymerase chain (RT-PCR) has also been upgraded with point-of-care rapid tests. Supportive treatment, mechanical ventilation, and extracorporeal membrane oxygenation (ECMO) remain the primary choice, while therapeutic options include antivirals, antiparasitics, anti-inflammatories, interferon, convalescent plasma, monoclonal antibody, hyperimmunoglobulin, RNAi, and mesenchymal stem cell therapy. Different types of vaccines such as RNA, DNA, and lentiviral, inactivated, and viral vector are in clinical trials. Moreover, rapidly deployable and easy-to-transport innovative vaccine delivery systems are also in development. As countries have started easing down on the lockdown measures, the chance for a second wave of infection demands strict and rational control policies to keep fatalities minimized. An improved understanding of the advances in diagnostic tools, treatments, vaccines, and control measures for COVID-19 can provide references for further research and aid better containment strategies. In late December 2019, the health officials in China reported of a mysterious pneumonia with no known etiology. Prompt genome analysis revealed that the disease was the cause of a novel coronavirus. As the outbreak continued to spread out at a remarkable pace, the World Health Organization (WHO) had to declare it a pandemic on March 11, 2020. SARS-CoV-2 is the seventh coronavirus to infect humans and the causative agent for COVID-19. Previously, two coronavirus epidemics raised international concern, SARS in 2002–2003 and MERS in 2012. As for the new virus, it is highly infectious and has already killed over 200,000 people with an estimated sCFR  of 1.4% (0.9–2.1%) [WHO recommends nucleic acid amplification test (NAAT) based reverse transcription polymerase chain reaction (RT-PCR) as the primary testing method because of its accuracy and hence it remains the gold standard for COVID-19 detection . HoweverWith overloaded healthcare and an increasing number of infections among the medical personnel, the ultimate way out remains to be in the discovery of an effective vaccine. However, the discovery and development of vaccine or drugs is a lengthy process, and it generally takes around a decade to pass through the entire course. So, repurposing of existing drugs to treat COVID-19 appears as a logical scientific approach. So far, the choices are limited for the treatment of COVID-19 by a lack of specific drugs. From a variety of existing antiviral medications, repurposing the appropriate drug remains as a challenge to overcome. Small-scale studies reported a few drugs to be effective, but later proved to bring no significant difference in clinical outcomes –8. High As the virus is moving in a pandemic speed, various control measures have been considered by different regions. Revising the decisions and their respective outcomes can further resolve the challenges in disease containment. In this review, we aimed at summarizing the current literatures to draw a compendium of understanding on the scopes of diagnostics tools, treatments, vaccines, and control measures for COVID-19.The real time reverse transcriptase polymerase chain reaction (rRT-PCR), or quantitative reverse transcriptase PCR (qRT-PCR), uses nucleic acid amplification test (NAAT) method to convert the viral RNA into cDNA and subsequently amplify the DNA for detection as a definitive confirmatory test . CurrentThe RT-PCR has a few disadvantages such as the requirement of strict handling, time consumption, and a minimum initial concentration of RNA. The lowest RNA concentration of SARS-CoV-2 detectable via RT-PCR is found to be 3.8–23 copies/ml . Heat trFor specimen collection, with regard to upper respiratory specimens, nasopharyngeal and oropharyngeal swab should be considered, and for lower respiratory specimens, sputum and endotracheal aspirate or bronchoalveolar lavage should be considered . AdditioSome point-of-care molecular testing methods have already been approved by emergency use authorization (EUA). For instance, Accula SARS-CoV-2 testing is a qualitative visually read PCR method that provides results in 30 minutes; Abbott ID NOW COVID-19 test uses an isothermal nucleic acid amplification technology to target the RdRp gene, and the positive result can be obtained as quickly as 5 minutes , 18.In addition, rapid test design for the detection of viral protein is also in progress. The viral nucleocapsid (N) and spike (S) are the main immunogenic proteins. While nucleocapsid protein is the most abundant and 90% similar to SARS-CoV, viral spike (S) protein is divergent and elicits strong immune response . HoweverDevelopment of an accurate antibody testing is a major challenge, and as of now, hundreds of trials are ongoing. There are important considerations for antibody testing, (i) timing of the test, (ii) previous infection, (iii) immune status of the individual, and (iv) cross-reaction, which can alter the test result. However, a few antibody-based tests got emergency approval by the FDA, for instance, COVID-19 ELISA IgG Antibody Test from Cellex, which has a sensitivity of 93%, and qSARS-CoV-2 IgG/IgM Rapid Test from Mount Sinai Laboratory with 92.5% sensitivity , 22. TheAlicyclobacillus acidiphilus. The test recently got FDA approval under emergency use authorizations [CRISPR gene editing tool has been employed to construct an accurate, faster, and simple-to-use SARS-CoV-2 detection test. DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTR) assay is based on CRISPR–Cas12 and it can distinguish SARS-CoV-2 with no cross-reactivity for related coronavirus strains using N gene gRNA within 40 minutes . The resizations . The tesizations . The assApart from detecting the presence of the virus, chest CT scan can demonstrate the disease status and severity. A recent study suggested that CT scan is more sensitive than the PCR procedure . During Recently, artificial intelligence system based on deep convolutional neural network design was used to detect COVID-19 from chest radiography images . The tooDifferential diagnosis is the process by which a single disease or condition is differentiated from those having similar clinical features. Patients with COVID-19 can have coinfection or superimposed infection by other viruses or bacteria simultaneously. Differential diagnosis is therefore important to differentiate SARS-CoV-2 induced infection from other viral or bacterial and mycoplasmal pneumonia. Since a plethora of clinical manifestations is observed in COVID-19, there is an increased chance for incorrect diagnosis. For instance, a COVID-19 patient in Thailand was found to present with fever and rash and was initially mistaken for dengue , 37. In For most patients with mild-to-moderate symptoms, bed rest with supportive treatments including sufficient calorie, water intake and maintaining water electrolyte balance, and homeostasis have shown to provide relief. However, for patients with hypoxia, a noninvasive positive airway pressure ventilator could be used. In case of disease severity, the patient is supported with invasive mechanical ventilation with endotracheal intubation. According to WHO-China joint report, about 25% of severe and critical cases require mechanical ventilation while the remaining 75% can be safely supplemented with oxygen only . With acRemdesivir, a prodrug that was previously intended to be used against Ebola virus, is now considered by the WHO as one of the most promising candidates to treat COVID-19 , 43. RecThe cocktail of danoprevir and ritonavir is in phase-IV trial indicating to be a promising therapeutic option. Danoprevir is an inhibitor of NS3/4A HCV protease while ritonavir acts to inhibit cellular CYP3A4 to bolster danoprevir's activity. A clinical study conducted in China described eleven patients gaining complete recovery after administering danoprevir boosted with ritonavir . Also, tAmong the other repurposable antiviral drugs, favipiravir is of notable consideration as it acts to selectively inhibit viral RNA polymerase enzyme against a broad range of viruses . The druNafamostat, a potent MERS-CoV inhibitor, was also found active against TMPRSS2-dependent host cell entry of SARS-CoV-2 . To prevSome anti-inflammatory drugs are also made use of to decrease disease severity. For instance, sargramostim is a recombinant GM-CSF that functions as an immunostimulatory agent and augments in innate host defense against pathogens . Among oOther potential repurposable drugs include steroids as anti-inflammatory agents. Although the use of methylprednisolone in extreme cases such as ARDS indicated a chance of lower mortality, the regular use of corticosteroids is not recommended as the effectiveness is yet to be confirmed . HoweverPreviously, preliminary studies indicated that chloroquine/hydroxychloroquine together with azithromycin could potentially reduce the disease severity of COVID-19 patients . In vitrRecently, an antiparasitic drug suramin was proposed as a possible therapeutic option as it was found to inhibit viral entry . The in β 1a could inhibit SARS-CoV in Vero E-6 cell line [β was proposed as an appropriate treatment choice. A study showed that IFN-I treatment caused significant reduction in viral protein and replication in SARS-CoV-2 [During SARS outbreak, interferons were used to treat patients, and laboratory experiment showed that IFN-ell line . For COVRS-CoV-2 . Howeverpro protein out of 1.3 billion compounds of ZINC15 library [The development of novel drug targeting the specific proteins of SARS-CoV-2 is also advancing. One study described deep docking method to identify potential 1000 ligands against the SARS-CoV-2 M library . Another library .Of different viral proteins, 3-chymotrypsin-like protease, papain-like protease, helicase, and RNA-dependent RNA polymerase, spike glycoprotein could be targeted for drug development . MoreoveRNAi or RNA interference could be used to construct novel drugs as the technique involves inhibiting gene expression by neutralizing targeted mRNA molecules. The RNAi was extensively explored against SARS-CoV, including the small interfering RNA (siRNA). One study reported that viral replication can be reduced up to 90% using siRNA-based RNAi technology in Vero E6 cells .Convalescent plasma (CP) treatment using blood from recovered COVID-19 patients could be used to aid in recovery for critically ill patients. A considerable number of clinical trials are underway to decide on the safety and efficacy of the treatment, and preliminary results were promising . A studyMonoclonal antibodies (mAB) specific to SARS-CoV are also being tried against SARS-CoV-2. One lab experiment reported that the human monoclonal antibody, CR3022, can bind to the receptor binding domain (RBD) of SARS-CoV-2 . AnotherThe development and production of hyperimmunoglobulin (H-Ig) is underway, which could be more potent and easily deployable than convalescent plasma . It contThe mesenchymal stem cell (MSC) therapy is also thought to be utilized as they can exert anti-inflammatory and anti-apoptotic effects along with repairing pulmonary epithelial cell damage as well as promoting alveolar fluid damage . ResearcThe development of vaccine is in progress, but it is estimated that a working safe vaccine will require at least 12–18 months to be available for wider use . There atuberculosis. However, there is no evidence so far to confirm that the BCG vaccine prevents SARS-CoV-2 infection in humans.The Ad5-nCoV is in phase-II in China and ChAdOx1-nCoV is preparing to enter phase-II trial in the UK. The primary result from the recombinant Ad5-nCoV vaccine expressing the spike glycoprotein showed to stimulate humoral response and the vaccine was found to be safe and tolerable . Other nLiquid-based intramuscular needle-and-syringe injections have a few caveats. They are expensive to store and transport; many of them are sensitive to temperature. Some advanced approaches are in active development for COVID-19 such as the Langerhans cell targeted delivery system (LC-TDS) . The LC-Other options that can be explored in vaccine delivery include solid dose vaccine delivery system based on mucosal route to present antigens to mucosa associated lymphoid tissues (MALT). The vaccines currently available via oral route include rotavirus, typhoid, cholera, and poliovirus. Moreover, a live attenuated typhoid vaccine Ty21a is available as orally delivered capsule that can be readily ingested . Fast diTo curb the crisis, the citizens are advised to follow basic guidelines such as, frequent hand washing, using disinfectants, following cough etiquette, and using facemask. Recent studies suggest that masks could slow down the disease transmission . CDC nowCOVID-19 is highly infectious, and contact transmission might occur due to touching the mouth, nose, or eyes with contaminated hands. On average, people tend to touch their faces every two and a half minutes, indicating how quickly the virus can make way into a human host . With thAccording to the European standard EN 149 + A1 : 2009, there are three levels of precautions for healthcare workers: (i) contact, (ii) droplet, and (iii) airborne precaution . MedicalTo contain the virus, a strict lockdown was imposed in Wuhan. Lockdown allows quick suppression of the number of infections facilitating time for the healthcare system to respond to the epidemic with planning and resource mobilization . One stuThe lockdown strategy and the complete closure of workplaces and termination of domestic and international flights was also argued. One study discussed that the decreasing trend of epidemic in Hubei, China, was possibly not driven by the internal travel ban or lockdown . AnotherHowever, in Brazil, a unique on-off lockdown strategy was undertaken through intermittent relaxation of lockdown . In the As countries around the world have started easing down on the previously imposed restrictions such as opening up businesses and shops and allowance for travel, there is an increased risk for a second wave of infection, which would be difficult to contain, especially during the winter season. Premature termination of strict measures could potentially lead to a second wave of infection. One modeling study demonstrated that relaxing the lockdown too early would cause the R0 to exceed 1 and spread across China. Epidemiologic disease modeling could play a vital role by predicting the plausible infection rate status. Modern technology such as machine learning could be used to process a large amount of data and generate better models .The continued infection spike across various countries despite lockdown measures indicated that the effectiveness of lockdown depended heavily on the proper testing, contact tracing, quarantining, and imposing strict physical distancing methods in high risk areas . Urban iAmidst the pandemic, there is an overflow of misinformation regarding the protection and cure from COVID-19. The infodemic of misinformation creates panic and poses a serious threat to public health attempts to contain the virus. Panic buying, false cures, and spread of disinformation amplify the risk of health loss and social disorder. The risk communication team of WHO launched a new platform called WHO Information Network for Epidemics (EPI-WIN) to provide credible information regarding COVID-19 . It is iThe pandemic of COVID-19 has challenged our existing knowledge, laws, and regulations and forced us to take measures as far as complete lockdown in various parts of the world. The high death toll of COVID-19 has stressed the need for prompt research and dissemination of updated information. This review summarized the scopes and developments of COVID-19 diagnosis tools and therapeutic options and discussed the prevention and control measures considering an apparently upcoming second wave of infection.While the world is in search for a cure, it is recommended that countries make use of existing scientific tools to develop models to predict community-based outcomes prior to making decisions. Healthcare workers must be supported with supplies and remain updated with the up-to-date knowledge, and citizens must play their role to maintain basic guidelines. At the governmental level, facilitating more testing and contact tracing, providing timely publication of epidemic information, enabling early diagnosis, and delivering supportive treatments for the patients are of utmost importance.


Studies with behavioural and neuropsychological tests have supported the developmental taxonomy theory of antisocial behaviour, which specifies abnormal brain development as a fundamental aspect of life-course-persistent antisocial behaviour, but no study has characterised features of brain structure associated with life-course-persistent versus adolescence-limited trajectories, as defined by prospective data. We aimed to determine whether life-course-persistent antisocial behaviour is associated with neurocognitive abnormalities by testing the hypothesis that it is also associated with brain structure abnormalities.We used structural MRI data collected at 45 years of age from participants in the Dunedin Study, a population-representative longitudinal birth cohort of 1037 individuals born between April 1, 1972, and March 31, 1973, in Dunedin, New Zealand, who were resident in the province and who participated in the first assessment at 3 years of age. Participants underwent MRI, and mean global cortical surface area and cortical thickness were extracted for each participant. Participants had been previously subtyped as exhibiting life-course-persistent, adolescence-limited, or no history of persistent antisocial behaviour  based on informant-reported and self-reported conduct problems from the ages of 7 years to 26 years. Study personnel who processed the MRI images were masked to antisocial group membership. We used linear estimated ordinary least squares regressions to compare each antisocial trajectory group (life-course persistent and adolescence limited) with the low trajectory group to examine whether antisocial behaviour was related to abnormalities in mean global surface area and mean cortical thickness. Next, we used parcel-wise linear regressions to identify antisocial trajectory group differences in surface area and cortical thickness. All results were controlled for sex and false discovery rate corrected.Data from 672 participants were analysed, and 80 (12%) were classified as having life-course-persistent antisocial behaviour, 151 (23%) as having adolescence-limited antisocial behaviour, and 441 (66%) as having low antisocial behaviour. Individuals on the life-course-persistent trajectory had a smaller mean surface area  and lower mean cortical thickness  than did those in the low group. Compared with the low group, the life-course-persistent group had reduced surface area in 282 of 360 anatomically defined parcels and thinner cortex in 11 of 360 parcels encompassing circumscribed frontal and temporal regions associated with executive function, affect regulation, and motivation. Widespread differences in brain surface morphometry were not observed for the adolescence-limited group compared with either non-antisocial behaviour or life-course-persistent groups.These analyses provide initial evidence that differences in brain surface morphometry are associated with life-course-persistent, but not adolescence-limited, antisocial behaviour. As such, the analyses are consistent with the developmental taxonomy theory of antisocial behaviour and highlight the importance of using prospective longitudinal data to define different patterns of antisocial behaviour development.US National Institute on Aging, Health Research Council of New Zealand, New Zealand Ministry of Business, Innovation and Employment, UK Medical Research Council, Avielle Foundation, and Wellcome Trust. Children and adolescents exhibiting persistent antisocial behaviour are often diagnosed with conduct disorder. These children are at an increased risk for incarceration and poor physical and mental health later in life.Evidence before this studyThe developmental taxonomy theory of antisocial behaviour outlines two prototypes of antisocial behaviour: life-course persistent and adolescence limited. We searched PubMed and Google Scholar for research articles published between Aug 1, 2018, and April 30, 2019, using the terms “antisocial behavio(u)r”, “conduct problems/disorder”, and “developmental taxonomy”, and we reviewed publications reporting structural brain measures. English language publications were included. Longitudinal studies have documented abnormal neuropsychological development in individuals with life-course-persistent but not adolescence-limited antisocial behaviour. However, no studies have reported on differences in structural brain integrity between these two groups defined by data-driven models.Added value of this studyThis study addresses limitations of existing research by examining a population-representative longitudinal birth cohort and by using statistically defined groups of individuals with life-course-persistent or adolescence-limited antisocial behaviour. Providing support for the developmental taxonomy theory, individuals on the life-course-persistent antisocial trajectory had thinner cortex and smaller surface area in brain regions associated with executive function, motivation, and affect, including the ventromedial prefrontal and orbitofrontal cortices, superior temporal gyrus and posterior cingulate cortex, than did individuals who were not antisocial, and these differences were more broadly distributed across the brain than previously reported in cross-sectional studies of antisocial individuals. Moreover, the adolescence-limited group had thinner cortex in the right middle and inferior temporal lobe, regions that have not been associated with antisocial behaviour previously.Implications of all the available evidenceThe developmental taxonomy theory has affected juvenile justice policy across the globe as well as clinical diagnosis and treatment. Our study supports the developmental taxonomy theory by showing that individuals on the life-course-persistent but not adolescence-limited trajectory show robust, widespread structural brain differences compared with individuals without antisocial behaviour.Large, well defined cohort studies indicate that individuals on the life-course-persistent trajectory display difficult childhood temperament and antisocial behaviour early in life.Adolescence-limited antisocial behaviour emerges alongside puberty and reflects socially normative peer processes, including gaining and asserting independence from adults.The original formulation of the taxonomy specified abnormal brain development as a fundamental aspect of the aetiology of life-course-persistent but not of adolescence-limited antisocial behaviour,We aimed to address the questions of whether previously reported structural brain differences linked to antisocial behaviour predominantly characterise individuals on the life-course-persistent trajectory, as hypothesised in the developmental taxonomy theory, and whether there are structural brain differences associated with life-course-persistent antisocial behaviour that have not been detected in previous small, cross-sectional studies. We focused on two structural features of neocortex—surface area and cortical thickness—making it possible to assess whether either of these measures was more indicative of antisocial behaviour pathology or differentially associated with antisocial behaviour trajectories. Our focus reflects both the preferential role of neocortical circuits in supporting higher-order executive processes implicated in neuropsychological deficits associated with life-course-persistent antisocial behaviourParticipants belonged to the Dunedin Study,The relevant ethics committees approved each study phase, and informed consent was obtained from all participants.Participants were scanned using a Siemens Skyra 3T scanner  with a 64-channel head and neck coil. As part of the scanning protocol, high-resolution T1-weighted images, three-dimensional fluid-attenuated inversion recovery (FLAIR) images, and a gradient echo field map were obtained. Acquisition parameters and analysis preprocessing details are described in the We analysed structural MRI data using the Human Connectome Project (HCP) minimal preprocessing pipeline, as detailed elsewhere.t tests and resulting p values were used to compare groups on demographic and cognitive variables. Odds ratios and associated CIs were calculated to compare groups on psychiatric diagnoses at 45 years of age. First, we used linear estimated ordinary least squares regressions to compare each antisocial trajectory group (life-course persistent and adolescence limited) with the low trajectory group to examine whether antisocial behaviour was related to abnormalities in global surface area and mean cortical thickness. Next, we used parcel-wise linear regressions to identify antisocial trajectory group differences in surface area and cortical thickness in each of the 360 regions comprising the parcellation scheme described previously.Participants who met MRI data quality control measures were included in analyses. Student's All analyses were done with R, version 3.4.1.The funders had no input in study design, data collection, data analysis, data interpretation, writing of the report, or the decision to submit for publication. All authors had access to study data. The corresponding author had final responsibility for the decision to submit for publication.vs low group; standardised β=–0·06 [95% CI −0·12 to 0·00], p=0·071, for the adolescent-limited vs low group; vs low group; standardised β=–0·08 [95% CI −0·16 to 0·00], p=0·039, for the adolescent-limited vs low group; The most recent phase of data collection was at 45 years of age, completed in April, 2019. Of the participants with available structural MRI data, four were excluded because of major incidental findings or previous injuries , nine because of missing FLAIR or field map scans, one because of poor surface mapping, and ten because of other quality control issues. Of the 672 participants who had both usable MRI data and were subtyped in trajectory groups, 80 (12%) were classified as having life-course-persistent antisocial behaviour, 151 (23%) as having adolescence-limited antisocial behaviour, and 441 (66%) as having low antisocial behaviour. These proportions resemble those originally identified within the full cohort, including individuals who did not undergo MRI .In the exploratory analyses, p values derived through permutation testing for these comparisons for both surface area and cortical thickness are similar to those reported in Secondary analyses including IQ, socioeconomic status, total intracranial volume, and head-injury history, and excluding individuals diagnosed with schizophrenia left findings largely unaltered .Our analyses revealed findings in line with the developmental taxonomy theory. First, we found that smaller surface area and thinner cortex in brain regions associated with executive function, motivation, and affect regulation previously implicated in studies of antisocial behaviour were, as hypothesised, specific to the life-course-persistent group. Second, the observed pattern of smaller surface area specific to the life-course-persistent group was much broader than previously reported in cross-sectional studies of antisocial behaviour. Third, both antisocial behaviour trajectory groups had features of reduced cortical thickness compared with the low group without antisocial behaviour, but in different areas of the brain. In the adolescence-limited group, the pattern was limited to two regions in the right temporal lobe that have not been consistently implicated in previous studies of antisocial behaviour, whereas in the life-course-persistent group, the pattern extended to paralimbic frontal and temporal regions that have been previously implicated in antisocial behaviour.So far, the majority of research comparing life-course-persistent and adolescence-limited antisocial behaviour (or proposed proxies for these subtypes) has not used optimal study designs for investigating the underlying brain correlates of antisocial behaviour subtypes as originally delineated by the developmental taxonomy theory.Early-onset antisocial behaviour, which characterises the life-course-persistent group, is strongly heritable, and the people with life-course-persistent antisocial behaviour are also more likely to have experienced childhood adversity.It is also possible that the observed structural brain features arose from a lifetime of confounding risk factors  and are therefore a consequence of the life-course-persistent antisocial lifestyle. Secondary analyses including IQ, socioeconomic status, total intracranial volume, and head-injury history, and excluding individuals diagnosed with schizophrenia left findings largely unaltered. It was not meaningful to separate these risk factors because they have been shown to be virtually inherent core characteristics of the life-course-persistent trajectory .6 ExcludSeveral explanations could account for the pervasive and non-specific signature of smaller surface area associated with life-course-persistent antisocial behaviour. Surface area and cortical thickness are under distinct genetic control,In contrast to the broad patterns of smaller surface area, lower cortical thickness associated with life-course-persistent antisocial behaviour was more circumscribed. Differences in the structure of the ventromedial prefrontal cortex and posterior cingulate have been previously shown in adults with persistent antisocial behaviourParticipants with life-course-persistent antisocial behaviour also had thinner cortex in the superior temporal gyrus and temporal pole than those without antisocial behaviour. Reduced thickness in the superior temporal gyrus of youths with conduct disorder has been reported previously and is hypothesised to be related to social-cognitive impairments that are pronounced in people with early-onset conduct disorder.The developmental taxonomy theory of antisocial behaviour, first posed 25 years ago, has influenced juvenile justice policy in the UK, USA, and elsewherehttps://moffittcaspi.trinity.duke.edu/research). We offer secure access on Duke University , University of Otago , and King's College London  campuses. All data analysis scripts and results files are available for review.The datasets reported in the current article are not publicly available because of lack of informed consent and ethical approval, but are available on request by qualified scientists. Requests require a concept paper describing the purpose of data access, ethical approval at the applicant's university, and provision for secure data access (


Following brain parcellation by using independent component analysis, dynamic CAP analyses were conducted and k-means clustering was used to determine transient activation patterns of the M-CIN. The frequency of occurrence of different dynamic CAP brain states was then compared between children with ASD and typically developing (TD) children. Dynamic brain configurations characterized by coactivation of the M-CIN with central executive/lateral fronto-parietal and default mode/medial fronto-parietal networks appeared less frequently in children with ASD compared with TD children. This study highlights the utility of time-varying approaches for studying altered M-CIN function in prevalent neurodevelopmental disorders. We speculate that altered M-CIN dynamics in ASD may underlie the inflexible behaviors commonly observed in children with the disorder.Brain connectivity studies of autism spectrum disorder (ASD) have historically relied on static measures of functional connectivity. Recent work has focused on identifying transient configurations of brain activity, yet several open questions remain regarding the nature of specific brain network dynamics in ASD. We used a dynamic coactivation pattern (CAP) approach to investigate the salience/midcingulo-insular (M-CIN) network, a locus of dysfunction in ASD, in a large multisite resting-state fMRI dataset collected from 172 children  is a neurodevelopmental disorder associated with altered patterns of functional brain connectivity. Little is currently known about how moment-to-moment brain dynamics differ in children with ASD and typically developing (TD) children. Altered functional integrity of the midcingulo-insular network (M-CIN) has been implicated in the neurobiology of ASD. Here we use a novel coactivation analysis approach applied to a large sample of resting-state fMRI data collected from children with ASD and TD children to demonstrate altered patterns of M-CIN dynamics in children with the disorder. We speculate that these atypical patterns of brain dynamics may underlie behavioral inflexibility in ASD. Autism spectrum disorders (ASDs) are neurodevelopmental disorders with central features of atypical social communication and restricted and repetitive behavioral patterns  ROI permits the identification of multiple different coactivation patterns, breaking down traditional static FC findings into multiple contributing patterns  produces dynamic CAPs that are less variable  than dynamic brain states identified using a sliding window correlation analysis –based FC map into multiple transient patterns occurring at different points in time. Liu and Duyn first showed that applying Compared with static FC methods, dynamic time-varying approaches may exhibit superior accuracy in characterizing the behavior of typically developing (TD) individuals, as well as in discriminating clinical from nonclinical populations including autism, schizophrenia, depression, and bipolar disorder  to resting-state fMRI data collected from three cohorts of age- and IQ-matched children with ASD and TD children and conducted a dynamic CAP analysis focusing on the M-CIN. The goals of the study were to (a) identify patterns of dynamic CAPs related to the M-CIN, and (b) compare these dynamic M-CIN coactivation patterns between TD and ASD groups. We hypothesized that children with ASD would show altered M-CIN dynamics compared with TD children.n = 75) and TD (n = 97) groups did not differ in terms of age (p = 0.285), full-scale IQ (p = 0.127), or mean head motion (p = 0.917). For participants from UM and Stanford University, ASD diagnosis was confirmed by a licensed psychologist using the Autism Diagnostic Observation Schedule (ADOS-2)  public databases  . Preprocessing steps included realignment, normalization to the 3-mm MNI template, and smoothing .As the three sites utilized different scan lengths, each scan was trimmed to 155 timepoints, which was the length of the data collected at the Erasmus site. We removed the first five volumes from the beginning of the scans to remove any scanner initiation artifacts, and removed additional volumes from the end of the scans, as needed, to make each scan 5 minutes and 10 seconds in duration. The scans were then subjected to quality control procedures as follows. All neuroimaging data underwent a visual quality control procedure conducted by trained research assistants to identify scanner- or motion-induced artifacts. Artifacts included, but were not limited to, signal loss, head coverage, motion slicing, ringing, blurring, ghosting, and wrapping. Data were rated as either pass, qualified pass, or fail. All images included in the analysis received either a pass or a qualified pass rating. The resting-state scans were then preprocessed using DPABI version 3.1 (We did not apply https://trendscenter.org/software/gift/). We used a model order of 100 independent components (ICs) as in previous similar work  . All 100 ICs were visually inspected and classified as noise or non-noise by two of the authors (JN & EM). The ICA components related to movement, white matter, or cerebrospinal fluid were removed from further analysis. The remaining 69 components were grouped into 12 functional domains to facilitate CAP interpretation: salience/M-CIN, default mode/M-FPN, central executive/L-FPN, sensorimotor, frontal, parietal, temporal, occipital, subcortical, attentional, cerebellum, and brainstem   × 100 (ICs) matrix for each of the 172 participants, and were postprocessed using Matlab code from the GIFT toolbox. Postprocessing included linear detrending, despiking using the AFNI 3D despike command, and bandpass filtering (0.01–0.1 Hz) Allen et al. .k-means clustering algorithm was applied to a concatenated matrix of all subjects’ timepoints of non-noise components to extract dynamic CAPs corresponding to different resting-state networks , mean head motion, age, and handedness.k as determined for all data. To determine if there was a relationship between specific CAPs, head motion and frequency of occurrence, average FD was computed for all TRs assigned to each CAP. Finally, to ensure that the identified CAPs were not driven by random differences in the data rather than brain activity, k-means clustering was conducted on phase-randomized surrogate data according to the procedure in Lancaster et al.    . State 2 0.0232) . Signifik-means clustering algorithm produced similar clusters in the ASD and TD participants, the above steps were run on the two groups, separately. There was no noticeable difference between the CAPs when the groups were clustered separately. The results of these analyses are detailed in To ensure that our There was no identifiable systematic relationship between head motion, the frequency of occurrence, or amount of TRs in each CAP . The mosFinally, phase-randomized surrogate data produced no noticeable CAPs of brain activity . The CAPASDs are widely thought to be associated with atypical patterns of functional brain connectivity  and two components in the default mode/medial fronto-parietal network (M-FPN). Children with ASD exhibited this CAP significantly less frequently than TD children. The lower frequency of occurrence of this state for children with ASD indicates less coactivation of the M-CIN with components of the M-FPN and L-FPN, especially the medial prefrontal cortex (mPFC) and PCC. Previous work has demonstrated that effective connectivity among nodes of these three networks can be used to discriminate task-evoked and resting states in TD children to a greater extent than in children with ASD. The same study found that this brain state discriminability was related to symptom severity in the domain of restricted and repetitive behaviors in children with ASD  was a top 10 component in States 1 and 3, and bilateral regions of the L-FPN were strongly activated in States 2 and 5. As activation of the L-FPN and M-FPN has been shown to be influenced and controlled by the M-CIN Uddin, , this coOnly one previous study has specifically examined anterior insula dynamics in ASD. Using a large ABIDE sample, Guo and colleagues identified an anterior insula ROI using Neurosynth , and attempted to control for acquisition site across all analyses, we cannot be sure that inherent between-site effects are completely accounted for. In addition, we cannot ascertain whether diagnostic procedures and scanner sessions were conducted in the same way for all participants at the three sites. There were also some discrepancies in the information that was reported to the ABIDE database for the different sites, eliminating the possibility of using behavioral measures like the Repetitive Behavior Scale or the Social Communication Questionnaire to explore relationships between dynamic FC parameters and symptom severity in the ASD group. Furthermore, similarly to most studies of ASD, our sample consisted mostly of males, with only 34 female participants included in the analyses. Although sex was included as a variable in our statistical analyses and had minimal effect on the significant results reported, this imbalance of males to females may fail to account for differences in the brain activity of the two sexes , Award ID: R01MH107549. Lucina Q. Uddin, Canadian Institute for Advanced Research. Lucina Q. Uddin, University of Miami Gabelli Senior Scholar Award. Jason Nomi, National Institute of Mental Health (http://dx.doi.org/10.13039/100000025), Award ID: R03MH121668. Jason Nomi, Brain & Behavior Research Foundation.Lucina Q. Uddin, National Institute of Mental Health (Click here for additional data file.