Circular RNAs (circRNAs) were known to be related to the pathogenesis of many diseases through competing endogenous RNA (ceRNA) regulatory mechanisms. However, the function of circRNA in coronary artery disease (CAD) remains unclear. In this study, we aim to construct a circRNA-related competing endogenous RNA (ceRNA) network in CAD.  The gene expression profiles of CAD were obtained from Gene Expression Omnibus datasets. Bioinformatics analysis was performed to construct a ceRNA regulatory network, from which the hub genes involved were identified through protein-protein interaction (PPI) networks leading to the identification of the circRNA-miRNA-hub gene subnetwork. In addition, function enrichment analysis was performed to detect the potential biological mechanism in which circRNA might be involved.  A total of 115 DEcircRNAs , 17 DEmiRNAs , and 790 DEmRNAs  were identified between CAD and control groups from microarray datasets. Functional enrichment analysis showed that DEmRNAs were significantly involved in inflammation-related pathways and ubiquitin-protein ligase binding. After identifying 20 DEcircRNA-DEmiRNA pairs and 561 DEmiRNA-DEmRNA pairs, we obtained a circRNA-miRNA-mRNA regulatory network. PPI network analysis showed that eight hub genes were closely related to CAD, leading to the identification of a circRNA-miRNA-hub gene subnetwork consisting of nine circRNAs , four miRNAs , and eight mRNAs . In addition, we discovered these hub genes were enriched in the ubiquitin-mediated proteolysis pathway, suggesting circRNAs may be involved in the pathogenesis of CAD through this pathway.  This study may deepen our understanding of the potential role of circRNA-miRNA-mRNA regulation network in CAD and suggest novel diagnostic biomarkers and therapeutic targets for CAD. Coronary artery disease (CAD), a cardiac disease caused by abnormal structure or function of the coronary artery, remains one of the leading causes of death and poses a heavy socioeconomic burden worldwide . PatientProtein-coding genes make up less than 2% of the human genome, while the majority of transcripts are noncoding RNAS (ncRNAs) . CirculaHere, in order to study the potential role of circRNA in the CAD, especially the circRNA-miRNA-mRNA regulatory network, we constructed a regulatory network consisting of 16 circRNAs, 6 miRNAs, and 267 mRNAs through bioinformatics analysis of multiple sets of the Gene Expression Omnibus (GEO) database. We used protein-protein interaction (PPI) networks to screen for hub genes and then constructed a circRNA-miRNA-hub gene subnetwork to further clarify the role of circRNA in CAD. Functional enrichment analysis was performed on these hub genes to understand the potential functional mechanisms of circRNAs. This study will provide in-depth insights into the potential pathogenesis of CAD and also suggest novel biomarkers and targets for CAD.http://www.ncbi.nlm.nih.gov/gds/). The circRNA expression data of CAD was obtained from GSE115733 [The microarray datasets used in the current study were acquired from the NCBI GEO database | >1 (|FC| > 2). DEmiRNAs were identified with the criteria of P value <0.05 and |log2FC| >0.263 (|FC| > 1.2).After downloading raw microarray data, normalization and logarithmic methods were performed to preprocess the data. The ID of the corresponding probe name was converted into an international standard name. When multiple probes correspond to the same gene symbol, the final probe expression value is calculated by the maximum value of probes. The probes that did not match a gene symbol were deleted. The Bioconductor Limma package was used to identify the DEcircRNAs , DEmiRNAs , and DEmRNAs  between CAD samples and the control samples. The criteria for selection of DEcircRNAs and DEmRNAs were https://david.ncifcrf.gov/). GO terms include biological process (BP), cellular component (CC), and molecular function (MF). A P value less than 0.05 was set as the cut-off criterion.To explore the potential biological mechanisms in the occurrence and development of CAD, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted by using the Database for Annotation, Visualization, and Integrated Discovery  [http://ophid.utoronto.ca/mirDIP/) [The circBank database (ank.cn/) , a comprmirDIP/) , as an iBy integrating the circRNA-miRNA pairs and the miRNA-mRNA pairs, we constructed a circRNA-miRNA-mRNA regulation network. The nodes that cannot complete a circRNA-miRNA-mRNA axis were removed. As circRNAs could competitively bind miRNA to regulate mRNA expression , only thhttps://string-db.org/) [The Retrieval of Interacting Genes/Proteins (STRING) database (db.org/) , an inteTo identify the association between DEcircRNAs, DEmiRNAs, and hub genes, we mapped the hub genes into the previously identified circRNA-miRNA-mRNA network and extracted the relevant DEcircRNAs and DEmiRNAs, based on which the circRNA-miRNA-hub gene subnetwork was identified. Cytoscape was used to visualize the circRNA-miRNA-hub gene subnetwork.A total of 115 DEcircRNAs (including 45 upregulated and 70 downregulated) and 790 DEmRNAs (including 496 upregulated and 294 downregulated) were identified from GSE115733 and GSE97320, respectively Figures . In addiGO and KEGG pathway enrichment analyses were performed on the 790 DEmRNAs identified in the previous section. The GO analysis demonstrated that the DEmRNAs were enriched in immune response (GO:0006955), inflammatory response (GO:0006954), and response to lipopolysaccharide (GO:0032496) for BP. When considering CC, hemoglobin complex (GO:0005833), cytosol (GO:0005829), and intracellular (GO:0005622) were the top three significantly enriched terms. With regard to MF, oxygen transporter activity (GO:0005344), ubiquitin-protein ligase binding (GO:0031625), and IgG binding (GO:0019864) were the most enriched terms. The top ten and total enriched terms are shown in Figures A total of 20 DEcircRNA-DEmiRNA pairs and 561 DEmiRNA-DEmRNA pairs were identified through bioinformatics prediction. As shown in The STRING database was used to construct a PPI network based on the 267 DEmRNAs with the unconnected nodes removed . The PPIBy mapping the eight hub genes into the previously identified circRNA-miRNA-mRNA network and extracting relevant circRNAs and miRNAs, a circRNA-miRNA-hub genes subnetwork was constructed. This subnetwork is consisted of nine circRNAs , four miRNAs , and eight mRNAs  . The basCAD is a complex and multifactorial disorder with high mortality and morbidity worldwide . The undκB) signaling pathway and the tumor necrosis factor (TNF) signaling pathway. It is well known that inflammation response plays an important role in the initiation and progression of CAD, as well as in coronary plaque instability [κB activation has been considered as a key step in the progression of atherosclerosis [α and NF-κB signaling pathways [α and NF-κB were significantly downregulated in CAD patients in Pakistan [In this study, a total of 115 DEcircRNAs, 17 DEmiRNAs, and 790 DEmRNAs between CAD and control groups were identified from three different microarray datasets in the GEO database, respectively. Moreover, we performed functional analysis of these DEmRNAs, and the results showed that these DEmRNAs were enriched in inflammation-related pathways such as the nuclear factor-kappa B  were identified in this subnetwork. The reports of these miRNAs have mainly focused on tumors. hsa-miR-136-5p was significantly downregulated in several types of cancers and functions as tumor suppressor. Zhu et al. revealed that hsa-miR-136-5p was associated with impaired tumorigenesis and metastasis in prostate cancer by targeting MAP2K4 . hsa-miRRecent studies have shown that the circRNA-miRNA-mRNA regulatory network could identify the disease-related circRNAs and potential axis . Miao etIn summary, our study constructed and analyzed a CAD-related circRNA-miRNA-mRNA network through integrated bioinformatics analysis, and we identified nine circRNAs that target the regulation of downstream genes through sponge-adsorbed miRNAs. However, there are still some limitations to this study. First, this is a retrospective study based on in silico data from the GEO database. All three datasets selected in this study were from different samples and different array platforms, which might have influenced the obtained conclusion. In addition, our results were mainly derived from bioinformatics predictions, and the proposed mechanism of circRNAs needed to be confirmed by laboratory studies. The advantage of our study is that it constructs a new circRNA-miRNA-mRNA network, which provides a new research direction for CAD. Based on this novel network, we further investigate the potential mechanism of circRNAs in CAD and identify potential biomarkers and therapeutic targets.In this study, we identified nine circRNAs  may be involved in the pathogenesis of CAD by constructing a circRNA-miRNA-mRNA regulatory network. Optimistically, this study may deepen our understanding of the potential pathogenesis of CAD and also suggest novel biomarkers and targets for CAD.


Recent observations raised concern that the intravenous recombinant tissue plasminogen activator (rt-PA) may result in damage to stroke patients caused by small artery occlusion (SAO). Thus, we perform a protocol for meta-analysis to investigate the efficacy and safety of intravenous thrombolysis with rt-PA in SAO-patients.The search-style electronic libraries, including Pubmed, Embase, the Cochrane Library, Web of Science, Wanfang Data, VIP Chinese Journals, and China Biomedical Literature Service System are used for document retrieval in June 2021 with no restrictions on language. The risk of bias in include articles will be assessed using the Cochrane Risk of Bias Tool. We perform the meta-analysis by Stata version 10.0 software and calculated the statistics using the inverse variance statistical method. Binary outcomes are presented as Mantel-Haenszel-style risk ratios with 95% confidence interval. Continuous outcomes are reported as mean differences.The results of the article will be shown in a peer-reviewed journal.Intravenous rt-PA may be effective and safe in SAO-patients. The response of treatment seems to be independent of underlying stroke mechanisms. Whether this is also true for stroke attributable to small artery occlusion (SAO) is under debate, because the presumed mechanism of SAO suggests that this stroke subtype may not respond to IVT.To reduce the burden associated with stroke, investigations of efficacy of available treatments for patients with stroke such as intravenous injection of recombinant tissue plasminogen activator (rt-PA) are necessary. In addition, patients with lacunar stroke treated by IVT have a poorer functional prognosis and higher risk of early deterioration than non-lacunar stroke patients treated by IVT. Moreover, leukoaraiosis, which occurs frequently in patients with SAO, has been confirmed to be a risk factor for the symptomatic intracranial hemorrhage after IVT. Thus, the bleeding risk related to IVT may be higher in SAO-patients when compared to patients with other stroke etiologies other than SAO. Recent observations raised concern that the use of rt-PA might cause harm in patients with strokes attributable to SAO. Thus, we conduct a protocol for systematic review and meta-analysis to investigate the efficacy and safety of intravenous IVT with rt-PA in SAO-patients.In the placebo-controlled randomized IVT trial, few patients suffered from stroke due to SAO, in spite of this cause accounted for 16% to 23% of all the strokes.2 Ethics application was not required as this study is based on published trials.This meta-analysis was registered at Open Science Framework registries (registration number: 10.17605/OSF.IO/ES39Z) and was conducted in accordance with the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols statement guidelines.2.1The search-style electronic libraries, including Pubmed, Embase, the Cochrane Library, Web of Science, Wanfang Data, VIP Chinese Journals, and China Biomedical Literature Service System are used for document retrieval in June 2021 with no restrictions on language. The search terms include “recombinant tissue plasminogen activator OR rt-PA,” “stroke OR cerebral infarction OR cerebral ischemia,” and “small artery occlusion.” These terms are searched in the combination of Medical Subject Headings (MeSH) and text words in the title and abstract. References to relevant review articles will be manually searched to identify additional studies2.2Studies are considered potentially eligible for this systematic review if they meet the following inclusion criteria: the objects were definitely diagnosed with stroke attributable to SAO; the intervention group was treated with intravenous rt-PA, while the control group was given placebo or conventional treatment; main outcome and complication measures were independence  at 3 months, intracranial hemorrhage, and recurrent ischemic stroke; randomized controlled trials. Studies would be excluded if they were available as case reports, letters, biochemical trials, conference abstracts, and reviews or if predefined outcome data required for analyses were lacking.2.3Two independent authors will extract the below descriptive information from the included articles: demographic information of patients, such as average age, number of patients, sex ratio and body mass index; study characteristics, such as author, year of publication, study language, study design, and the average follow-up period, and details of interventions and outcome measures. If the data cannot be directly extracted or is missing, we will contact the relevant author to ensure that the information is complete. Otherwise, we will calculate them with the guideline of Cochrane Handbook for Systematic Reviews of Interventions 5.1.0.2.4We use the Cochrane Collaborations risk of bias tool to assess the methodological quality and risk of bias of identified studies. Seven specific domains are evaluated in this tool: sequence generation, allocation concealment, blinding of participants and personnel, blinding of outcome assessment, incomplete outcome data, selective outcome reporting, and other issues. Each domain could be assigned as low risk of bias, high risk of bias, or unclear risk of bias based on the judgment criteria. To evaluate publication bias, we perform a funnel plot if the number of included studies is sufficient (>10 articles). A symmetrical funnel plot indicates no possibility of publication bias, whereas an asymmetrical funnel plot indicates a high possibility of publication bias. If we identify publication bias through analysis of the funnel plot, we may discuss possible reasons such as small-study effects.2.5I2 statistic. A random-effect model is adopted when I2 > 50% or P < .1; otherwise, heterogeneity is negligible and a fixed-effect model is used.We perform the meta-analysis by Stata version 10.0 software and calculate the statistics using the inverse variance statistical method. Binary outcomes are expressed as Mantel-Haenszel-style risk ratios with 95% confidence intervals. Continuous outcomes are presented as weighted mean differences (WMDs). Heterogeneity among the studies is quantified with the 3 SAO is a major ischemic stroke subtype that occurs more commonly in Asian than Western populations. However, to date, no evidence has shown the effectives of intravenous rt-PA for patients with stroke attributable to SAO. We acknowledge some of the limitations of this study. First, few studies have reported relevant topic and the sample size may be small; secondly, sometimes it is hard to distinguish different subtypes of ischemic stroke, which may affect the results of this study; thirdly, due to the low quality of the included studies, the evidence level may be declined. To draw more robust evidence, further randomized controlled trials are required.To the best of our knowledge, this is the initial meta-analysis to investigate the efficacy and safety of intravenous IVT with rt-PA in SAO-patients. Ischemic stroke is characterized by highly differentiated subtypes.Conceptualization: Li-Yan Ni.Formal analysis: Li-Yan Ni.Funding acquisition: Ji-You Tang.Investigation: Li-Yan NiMethodology: Ji-You Tang.Study design: Li-Yan NiWriting – original draft: Li-Yan NiWriting – review & editing: Ji-You Tang.


Built environment constructs derived from individual-level street network measures  were obtained. We assessed PA between four groups, based on income and GIS walkability . Results: No relationships between self-reported MVPA and GIS-measured walkability were found. Only intersection density was significantly, inversely associated with moderate and total MVPA . In the high SES group, vigorous PA was inversely associated with intersection density . Self-report transport PA differed between groups (p < 0.013). Conclusions: Results suggest that the construct of walkability may relate to volitional (leisure) and utilitarian (transport) PA differently, in highly inequitable settings.There is limited data concerning the built environment and physical activity (PA) in a country with a history of sociopolitically motivated, spatial and economic disparities. We explored the extent to which objectively measured attributes of the built environment were associated with self-report or device-measured PA in low- and high-socioeconomic status (SES) communities. Methods: In a convenient sample of residents ( The burden of physical inactivity is substantial in the country of South Africa  as it grThe monetary and spatial inequality trends in South Africa may only be addressed through policy interventions and solutions, targeting issues of social and environmental justice . The ecoOne key aspect of this ecological model is the built environment, which may be shaped by various planning and design processes, with the general targets for policies and structural interventions concerning physical activity usually found in the domains of “active travel” and “recreation or leisure time”. This study mainly focused on these two domains. Many studies have documented the relationship between supportive attributes of the built environment and physical activity for transportation and recreation purposes , yet mosIn one of the few studies in Africa, Malambo et al.  evaluateThe aim of this study, therefore, was to ascertain whether the objectively measured attributes of the built environment were associated with either self-reported or device-measured physical activity in an urban South African setting. Furthermore, we were interested in whether these relationships, should they exist, differ in high- and low-income settings, or by domains of activity.Materials: A cross-sectional survey design was used to collect information on physical activity and the neighborhood environment from community areas chosen to maximize the variability in neighborhood walkability (low-socioeconomic status (SES)/low walkable, high-SES/low walkable, low-SES/high walkable, high-SES/high walkable).The research study was conducted in four suburbs in the Western Cape Province of South Africa, namely, Langa, Pinelands, Khayelitsha and Table View, and all were in the urban metropole, South Africa. The suburbs were selected using clustered sampling method. Langa and Khayelitsha represented two primarily low socioeconomic status (SES) suburbs, whereas Pinelands and Table View represented suburbs of a higher SES, based on the city development index (CDI) that averaged indices of infrastructure, health, education and income. The suburbs of Khayelitsha and Langa have an average CDI of 0.75 compared to the provincial average of 0.81, whereas the suburbs of Table View and Pinelands have a CDI above 0.90. The four suburbs can be compared, as they are mainly residential, but have various commercial centers with retail, business, recreational facilities and public open space, with varying degrees of land use mix.n = 13), Table View (n = 13), Khayelitsha (n = 12) and Langa (n = 14).This study used a convenience sampling to select 66 adults (aged between 18 and 65 years) from who were regular members of either church or community groups in their respective suburbs for at least three months. Out of this sample, 14 of them had incomplete data, leaving us with a total of 52 adults with complete data needed for our analysis. The 52 included 10 men and 42 women; from Pinelands , high-SES/low walkable (n = 13), low-SES/high walkable (n = 13) and high-SES/high walkable (n = 13)). Approval for the study was granted by the University of Cape Health Sciences Research Ethics committee (HREC REF 293/2016).The sample was further divided into four groups using a median split for the GIS-measured walkability index (in 1000 m buffer) and by the neighborhood income level. The four study areas selected in Cape Town metropolis are shown in Compared to the rest of LMICs, Abubakar and Doan , show thWillemse and Donaldson , underscTheir research sought the perceptions, preferences, needs and uses of CNPs in five black townships in Cape Town including . Their research revealed that these townships had few CNPs, which therefore entailed travelling greater distances by public transport for access. Furthermore, the lack of private garden space forced the respondents to visit the CNPs and spend more time there thereby participating in either active or passive recreation. The main concerns for the CNPs included safety, maintenance and a lack of CNP facilities.The international physical activity questionnaire (IPAQ-long) English version was used to measure participants’ self-reported physical activity. This instrument has been previously validated in adults in South Africa with acceptable correlations . AdditioThe 31-item long form comprehensively assessed the frequency, duration and intensity of physical activity in the four domains of work, household, transportation and leisure. The IPAQ is used to compute the weekly dose of both moderate and vigorous physical activity . We did not translate and back translate the IPAQ into the South African local language spoken in Cape Town. Rather, we had trained local research assistants to administer the survey using local language equivalents of the words to match the intensities.www.ipaq.ki.se, accessed on 10 March 2021).We specifically examined walking or cycling for transportation and walking and physical activity for recreation. Work-related physical activity was not included, as it is not typically associated with the neighborhood built environment . TherefoParticipants were fitted with accelerometers  worn on an elastic belt around their waist. They were asked to wear them for seven days, and only to remove them when bathing or when they went to bed at night. Data were downloaded and analyzed using Actilife 6:10:4 software .The Actigraph GTX3 activity monitor was used to assess physical activity objectively. Minute by minute activity counts were accumulated and collapsed into minutes spent at different physical activity intensities, using the Freedson cut-points across the seven days . These iThe accelerometer data were collected and aggregated to one-minute epochs. Non-wear time was determined as any period of 60 min or more of consecutive zero counts. Only data from participants with at least 10 h of valid wear time, on at least four days, were included. Counts/minutes were converted into minutes of sedentary time (≤100 counts/min), light, moderate and vigorous-intensity physical activity as previously described .Measured built environment (GIS):Geographic Information Systems (GIS):Using geographic information systems (GISs) (ArcGIS version 10.51), we identified the available physical activity facilities, which included sporting venues, recreational centers and parks within a 1000 m residential buffer. The source of the point data was the City of Cape Town 2011 census. A radial buffer (1000 m) was established ,19 arounThe methodology used was based on Adams et al. , using tWalkability measures using GIS:Walkability:A walkability index was computed as the sum of Z scores for net residential density, land use mix and diversity and intersection density . The walThis score was adapted from the original measure, which included the retail floor area ratios, which were not available in the present study, but   capturedNet Residential Density:Net residential density was computed as the number of dwellings (numerator) divided by the land area dedicated to residential use, within the 1000 m buffer . The resStreet connectivity:In this study, , street Land-Use Mix and Diversity:Four land uses were computed viz. residential, retail-combined, civic/institutional and others. Parcel data was used to quantify the land uses. Land-use mix was calculated using an entropy equation  to scoreStatistical Analysis:t-tests. Where data were not normally distributed, for device-measured and self-reported physical activity, medians and the lower and upper quartiles were provided in addition to non-parametric Mann–Whitney U tests.Descriptive analysis of participants’ sociodemographic characteristics was undertaken and presented as means ± standard deviation for continuous variables or counts and percentages for categorical variables. The self-reported and device-measured MVPA were also compared between high- and low-SES groups using ANOVA. Between group comparisons for normally distributed were made using independent p < 0.05. All data were analyzed using Stat Soft® 2014 version 13 for Windows .The sample was divided into four groups using a median split for the walkability index . GIS-measured attributes that comprise the measure of walkability were calculated for the 1000 m buffer zones surrounding the participants’ street address. Spearman’s rho correlations were calculated between the reported physical activity for transport and recreation (IPAQ), device-measured physical activity and GIS measures of walkability. A Kruskal–Wallis test was used to compare self-reported and objectively measured physical activity between the four groups, according to SES and walkability. Statistical tests were considered significant at Descriptive characteristics of the participants:n = 52) are presented in p < 0.05) between the two-SES groups. Of those participants in the high-SES neighborhoods, 88% were either married or living with a partner, compared to only 19% from the low-SES communities. Additionally, 92% of persons surveyed in the high-SES suburbs compared to 44% from low-SES neighborhoods had access to at least one private motor vehicle.The descriptive characteristics of the participants for whom there was complete data  and transport physical activity (TPA) and total physical activity (TTL PA) were statistically significantly different. These differences are due to chance or lack of statistical power.Self-reported and device-measured physical activity and GIS walkability:p < 0.05). When groups were combined, the inverse relationship between intersection density and physical activity persisted. In the 1000 m buffer, intersection density was inversely associated with both moderate  and moderate-to-vigorous physical activity .There was no relationship between any self-reported physical activity and GIS measured attributes of walkability in 1000 m buffer as shown in Self-report and objectively-measured physical activity (MPVA) and GIS walkability when apportioned by income:p = 0.036) with significant between group differences between high-SES/low walkable vs. low-SES/high walkable (p = 0.013) as reported in p = 0.016), with between group differences for the low-SES/low walkable vs. high-SES/low walkable groups (p < 0.04).There was a significant overall difference in self-reported physical activity in the domain of transport  and device-measured physical activity (MVPA), in groups apportioned according to income and GIS measured walkability within a 1000 m buffer. In general, there was no relationship between self-reported physical activity and walkability , irrespective of income level. Conversely, the device-measured physical activity for all groups was inversely associated with intersection density. There were also observed differences in transport-related, self-report physical activity between SES groups.Previous studies have established that adults walk more for transportation in walkable neighborhoods . HoweverA vast majority of the literature has focused on examining associations between aspects of the built environment and modes of choice ,30,31. TBased on GIS and other measures, not all socioeconomic groups may benefit equally from the current notion of “walkable” built environments . StudiesThis study also points to socioeconomic inequalities, cultural and contextual differences in the environment , which are common to low- and middle-income countries, where PA is used for utilitarian, rather than recreational purposes. There is also the issue of environmental justice. The effect of environmental and social injustice still plagues South Africa post-1994. The skewed spatial legacies are indicated to determining opportunities in South Africa until now . The posStrengths and limitations:A strength of the present study was in its design to recruit participants from four different neighborhoods, based on income or socioeconomic status and residential density in an urban South African setting. The other strength included the use of accelerometers and GIS to objectively assess PA, and the perceived and objectively-measures of built environment. This study is one of the first to do so in Africa.The study has fundamental limitations to note. Conducting the study among residents of one neighborhood type or as part of a convenience sample (as participants in one area attend the same church within the same community) may restrict environmental variability. Restricted variability could, in turn underestimate the strengths of environmental–physical activity associations in environmental studies ,20,26,41The results of this study showed a mismatch between physical activity and the objectively measured attributes of the built environment that have been previously associated with “walkability”, largely in high-come countries. The findings are significant, especially given the high social disparities that face South Africa. This study suggests that persons living in low-income settings may not experience all the benefits of living in a walkable neighborhood, particularly if physical activity that is undertaken in utilitarian and not volitional. There is a need to consider the constraints to physical activity in low SES groups, along with the built environment, in order to address social and environmental justice in the promotion of physical activity, at a population level.The findings of this study could be relevant for health promotion and policy advocacy in sub-Saharan Africa, particularly in settings with a history of social and politically motivated, spatial disparities and inequities. Understanding the environmental correlates may lead to better strategies to promote physical activity in the low-income setting. Further evaluation of the relationship between physical activity and the built environment is needed in the broader South African population to strengthen evidence-based recommendations for creating better and safer communities with the built environment that improve access to safe and enjoyable opportunities in the African region.


Recent studies have shown that several children diagnosed with COVID-19 have developed Kawasaki Disease (KD)-like symptoms. This systematic review aims to assess the demographic, laboratory, and clinical characteristics of children with KD-like syndrome during the COVID-19 pandemic and evaluate efficacy of treatments and patients' outcome. A comprehensive search was carried out systematically through PubMed, Scopus, and Web of Science (WoS), medRxiv, and bioRxiv by two reviewers independently for all studies or preprints data on the demographic, laboratory, and clinical characteristics of children with K.D-like signs during the COVID-19 outbreak. Overall, 378 studies were identified by the systematic search, of which 25 studies were included in the study. The included studies involved 599 patients in total. Thirteen studies (52%) were case reports or case series, and the rest (48%) were cohort studies. In 19 studies, patients were diagnosed with Multisystem inflammatory syndrome in children (MIS-C). In 16 studies COVID-19 was diagnosed in all patients based on their polymerase chain reaction result, serological findings, and computed tomography results. Higher C-reactive protein and erythrocyte sedimentation rate level were the most prevalent laboratory findings. In most studies, patients had leucopenia with marked lymphopenia, hypoalbuminemia, and increased ferritin, as well as hyponatremia. Abnormal echocardiography and respiratory outcomes were the most common clinical outcomes. In 11 studies, all patients required intensive care unit admission. Findings of the present systematic review show that the incidence of KD-like syndrome in the COVID-19 pandemic increased significantly. Moreover, this study offers new insights in the KD-like syndrome pathogenesis and clinical spectrum during COVID-19 pandemic. Coronavirus Disease 2019 (COVID-19), caused by SARS-CoV-2, has created a global pandemic. Millions of people have been infected, and thousands have lost their lives  is one of the vasculitis of medium-sized vessels, that only affects children. Although no identified etiology can clearly explain this disease, some assumptions consider viral infections such as coronavirus family act as a trigger in genetically predisposed children , 8.Recent studies have shown that several children diagnosed with COVID-19 have developed symptoms such as prolonged fever, bilateral conjunctival injections, changes in the lips and oral cavity, cervical lymphadenopathy, extremity changes, and polymorphous rash that are similar to those of KD or Hyper inflammatory syndrome . On the This study is reported based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines , 13.The following inclusion criteria were used: (I) Published in the English language; (II) Full-text available; (III) observational studies (including case reports and case series); (IV) studies reporting the characteristics and outcome of children with KD-like syndrome during the COVID-19 outbreak; (V) patients in the study fulfill the classical Kawasaki criteria according to the American Heart Association indications (2017)  or KD-like syndrome criteria, based on CDC recommendation .A comprehensive search was carried out systematically through PubMed, Scopus, and Web of Science (WoS), databases , and medRxiv, bioRxiv  by two reviewers independently.*covid-19*” OR “*covid*” OR “*SARS-CoV-2*” OR “*2019-nCoV*” OR “*novel coronavirus*” OR “*new coronavirus*” OR “*coronavirus*”) AND (Kawasaki OR KD OR Kawasaki-like OR multi-system OR multi-system) AND inflammatory AND (syndrome* OR disease) OR MISC OR MIS-COR PIMS OR [inflammatory AND (multi-system OR multi-system) AND (syndrome* OR disease*) OR Systemic Inflammatory Response Syndrome OR hyperinflammatory] AND (child* OR pediatric).A search study was designed comprising of three concepts, the 2019 novel coronavirus disease, KD disease, and child. The following algorithm was used for screening the title and abstract ; . They came to an agreement regarding the conflicting results.Two independent researchers filled data extraction forms containing age, gender, journal, study type, sample size, clinical characteristics, laboratory findings, and outcomes. Another researcher resolved conflicts.Quality assessment (QA) of case-reports/case-series studies was assessed using the case-report (CARE) 13-item guideline . This guOur searches yielded 215 studies from PubMed, 310 studies from Scopus, and 156 study from Web of science. After removal of duplicates, we assess 378 studies for eligibility. We excluded 270 studies based on their title and abstract. Finally, 25 studies included in the qualitative synthesis. The detailed flow diagram is shown in In 16 studies COVID-19 was diagnosed in all patients based on their PCR result, serological findings, or computed tomography (CT) results. Fourhundred-fifty-five patients had positive serological indices  for SARS-CoV-2 infection, while 175 patients were tested positive for COVID-19 using PCR .Twenty one studies showed the CRP level; CRP was increased in all patients who participated in all studies. Highest CRP was reported in Deza Leon et al. study (450 mg/L) . A higheTwenty-two studies reported echocardiography findings of their patients. In six studies, all patients had abnormal echocardiography. The lowest rate of abnormality in echocardiography is demonstrated in Whittaker et al.  study 3. The mosIn this study, the highest and the lowest score of included case-reports/case-series based on CARE guidelines was 93 and 74%, respectively. The cohort studies included in this study were considered as high quality in seven articles and medium quality in five articles. This scale classifies papers as high quality (8–9 points), medium quality (6–7 points), and low quality (<6 points) .KD is a vasculitis of early childhood, which is the most common reason for acquired heart disease in children in developed countries . AlthougIn the Dean et al. study in a KD epidemic in Hawaii, 44 percent of the cases had a history of respiratory infection in the month before their visit. In their research, although an infectious agent is proved as one of the immune system triggers and an etiology for KD, yet the analysis failed to determine the microorganism responsible for it . Other sMembers of the coronaviridae family have also been suggested as triggers for KD . Some stWith the incidence of the COVID-19 pandemic, the number of children who presented with KD symptoms increased dramatically. The Verdoni et al. study showed that the monthly incidence of KD in an Italian province had increased 30-folds . The sigThis review has gathered the findings of 25 articles that have presented patients with KD-like syndrome in the era of the COVID-19 pandemic. Most of the patients have exhibited traces of COVID-19 in their tests. Among these data, it seems that KD-like syndrome are more strongly associated with positive serology tests. To justify this finding, we can point out that PCR turns negative in a shorter period compared to serology (measuring immunoglobulin levels) tests . It seemIn most reviewed articles, leukopenia and especially lymphopenia decreased levels of serum albumin, and an increase in Ferritin, ESR, and CRP was noted. Impaired heart function and myocarditis are the unfortunate outcomes of KD-like syndrome , 69. TheKD is rare disease, which was mainly managed by pediatric rheumatologists, but as a result of COVID-19 the prevalence of this disease and KD-like syndrome has increased notably and these disease spectrums are now considered as a health issue. Although most of the articles reviewed in this study have similar findings, this systematic review assists clinicians to determine the best therapeutic approach for their patients based on their demographic, clinical, and laboratory findings to achieve the best outcome.Although the clinical findings of KD is very similar to KD-like syndrome, there are some differences including the older age of KD-like Syndrome patients, which could be justified due to the higher exposure of school-aged children and adolescents to the virus compared to infants and toddlers. In addition, gastrointestinal Symptoms, lymphopenia, thrombocytopenia, and hypertriglyceridemia are more prevalent in KD-like syndrome compared to KD. KD-like syndrome patients are at higher risk of complications compared to KD patients. Moreover, the inflammatory factors have had more noticeable changes in these new patients, which may be related to primary infection with SARS-CoV-2 rather than KD itself , 71 Tab.It should be noted that there are some diagnostic limitations due to the relatively low sensitivity of serological test compared to PCR. Moreover, most of the articles that were reviewed in this study were case reports and had relatively low evidence. We found only a few articles concerning our subject (Even with lower evidence level). It is necessary to perform pathophysiological assessments in addition to conducting case-control studies so that we would be able to understand the relationship between the novel coronavirus and KD better.Incidence of children presenting with a severe inflammatory syndrome with KD-like features are increased during the COVID-19 pandemic. These children present with more severe symptoms which is attributed to worse clinical outcome and require intensive treatment and close monitoring. This study indicates that serological indices for SARS-CoV-2 have stronger correlation with the KD-like disease incidence compared to PCR. Moreover, CRP could also be helpful in terms of diagnosis. Although only one patient in included studies died of the complications the disease, most of the patients required ICU admission.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.MQ and KP: conceptualization. ME and MQ: search. PM and MA: data extraction. PM, ME, and MQ: writing—original draft. PI and KP: writing—review & editing. All authors read and approved final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Klebsiella pneumoniae (CR-Kp) and Acinetobacter baumannii (CR-Ab) represent important cause of severe infections in intensive care unit (ICU) patients. N-Acetylcysteine (NAC) is a mucolytic agent with antioxidant and anti-inflammatory properties, showing also in-vitro antibacterial activity. Aim was to evaluate the effect on 30-day mortality of the addition of intravenous NAC to antibiotics in ICU patients with CR-Kp or CR-Ab septic shock. A retrospective, observational case:control study (1:2) in patients with septic shock caused by CR-Kp or CR-Ab hospitalized in two different ICUs was conducted. Cases included patients receiving NAC plus antimicrobials, controls included patients not receiving NAC. Cases and controls were matched for age, SAPS II, causative agent and source of infection. No differences in age, sex, SAPS II score or time to initiate definitive therapy were observed between cases and controls. Pneumonia and bacteremia were the leading infections. Overall, mortality was 48.9% . Independent risk factors for mortality were not receiving NAC (p = 0.002) and CR-Ab (p = 0.034) whereas therapy with two in-vitro active antibiotics (p = 0.014) and time to initial definite therapy (p = 0.026) were protective. NAC plus antibiotics might reduce the 30-day mortality rate in ICU patients with CR-Kp and CR-Ab septic shock.Carbapenem-resistant  Klebsiella pneumoniae (CR-Kp) and Acinetobacter baumannii (CR-Ab) represent nowadays an important cause of severe infections in intensive care unit (ICU) patients and mortality rates are significantly associated to septic shock .p = 0.01), CR-Ab infection (p < 0.001), not receiving NAC (p = 0.05), whereas time to initiate definitive therapy (p = 0.017) and definitive therapy with two or more antibiotics displaying in vitro activity (p = 0.005) were protective.At the univariate analysis, risk factor for mortality were age (p = 0.002) and CR-Ab infection ; whereas time to initiate definitive therapy  and definitive therapy with two or more antibiotics displaying in vitro activity  were protective, regardless of age, sex, SAPS II score, source of infection or the type of antibiotics used as definitive therapy  was lower than that used in previous study, thus reducing the risk of toxic intermediates production.Finally, a recent randomized clinical trial investigating the effect of different anti-oxidants in septic shock showed that NAC was able to improve antioxidant capacity, in the absence of significant adverse reactions or side effects .Early antibiotic therapy represents a cornerstone of critical care management in septic shock patients ,44,45,46Although a difference was observed in the two study groups regarding the use of regimens containing colistin and tigecycline, with the former more frequently used in cases than controls, this did not affect overall mortality.In our study, 30-day mortality for CR-Ab was higher than that observed for CR-Kp. The impact of CR-Ab on clinical outcome was also highlighted in multivariable analysis, thus confirming the recent literature data, which showed a mortality rate of up to 60% and 40% in the presence of septic shock caused by CR-Ab and CR-Kp, respectively ,6,48.Our study has some limitations that should be acknowledged. First, the retrospective nature of the study is an intrinsic limitation of this analysis. Second, the sample size is relatively low and included all patients with septic shock caused by CR-Kp or CR-Ab observed in the two ICUs, without a sample size calculation. Therefore, the results of our study might be considered preliminary and further multicenter prospective studies are needed to confirm our findings. Third, cases and controls came from two distinct cohorts of patients and therefore differences could be related to intrinsic differences between populations and microorganism, despite the extensive patients’ matching . However, we were confident that the potential differences were minimized by the fact that both ICUs had a dedicated Infectious Diseases consultant who was in charge for the treatment of all patients and who belonged to the same well-established consultation system. Consequently, the Infectious Diseases consultants had the same diagnostic and therapeutic approach towards patients admitted to these 2 ICUs . Fourth,Nevertheless, we believe that the present investigation has some important strenghts, which might have contributed to bias reduction, such as: (i) patients were matched for several variables, which might have had theirselves an influence on the primary outcome of the study ; (ii) each ICU had a dedicated infectious diseases consultant referring to the same well-established consultation system, thus assuring the same clinical and therapeutic approach to infections in both cases and controls. Furthermore, it represents a real-life clinical experience providing useful suggestions to clinicians about the management of a difficult-to-treat infection such as septic shock caused by CR-Kp and CR-Ab.We performed a retrospective, observational case:control study (1:2) in patients with septic shock caused by CR-Kp or CR-Ab hospitalized in two different ICUs [IRCCS Neuromed for cases and Sapienza University  for controls, the latter derived from a historical cohort of patients . Both ICThe Ethical Commitees approved the study  whereas informed consent was waived due to the retrospective nature of the research.Patient data were extracted from medical records and from hospital computerized databases or clinical charts. The following information was reviewed: demographics, clinical and laboratory findings, comorbid conditions, microbiological data, duration of ICU and hospital stay, incidence of infections during hospitalization, treatments and procedures administered during hospitalization and/or in the 90 days prior to infection, classes of antibiotics received on admission and/or after admission before a positive culture of a biological sample was obtained, the simplified acute physiology score II (SAPS II) at the time of infection, source of infection, antibiotic regimens used for CR-Ab or CR-Kp infections, and 30-day mortality. According to both hospital’s guidelines, colonization with CR-Kp and CR-Ab strains was routinely evaluated by rectal swab, respiratory specimens and urine culture at the time of ICU admission and every week afterwards.Infections were defined according to the standard definitions of the ECDC  and septA CR-Ab or CR-Kp infection was defined as clinical signs of infection and culture of blood, urine, cerebrospinal fluid or a biological sample from skin and skin structures, lung, or abdomen yielding a CR-Ab or a CR-Kp strain.Infection onset was defined as the date of collection of the index culture . Infections were classified as hospital acquired if the index culture had been collected > 48 h after hospital admission and no signs or symptoms of infection had been noted at admission. Primary bloodstream infection (BSI) was defined as BSI occurring in patients without a recognized source of infection.The severity of clinical conditions was determined by using SAPS II score calculated at the time of septic shock onset. Length of hospital and ICU stay were calculated as the number of days from the date of admission to the date of discharge or death.Depending on the number of drugs used (one or more), treatment regimens were classified as monotherapy or combination therapy. Definitive antibiotic therapy was defined as the definitive antimicrobial treatment based on in vitro CR-Ab or CR-Kp isolates susceptibilities. Antibiotic regimens were also classified according to the following: one antibiotic displaying in vitro activity, and two or more antibiotics displaying in vitro activity. Time to initial definitive therapy was the time between infection onset and initial definitive therapy.Intravenous NAC was administered in patients with respiratory conditions characterized by excessive and/or thick mucus production as soon as signs of a possible infection developed  as adjunctive therapy and stopped together with antibiotic therapy. Dosages of intravenous NAC ranged from 1200 to 3000 mg/die, according to treating physicians. Intravenous NAC was administered in saline solution with 30–60 min infusion rate.p value two-sided <0.05 were considered statistically significant. The results obtained were analyzed using a commercially available statistical software package .Continuous variables were compared using Student’s t test or Mann-Whitney U test and were described as mean ± standard deviation (SD) or as median and interquartile range (IQR) according to whether the distribution of the variables was normal or non-normal. Chi-squared test (χ2) and Fisher’s exact test were used to compare categorical variables. Univariate and multivariate analyses were performed to evaluate factors related to 30-days mortality. Variables with a In conclusion, in the challenging context of increasing antimicrobial resistance and restricted therapeutic options, this study suggests that a combined use of NAC plus antibiotics might reduce the 30-day mortality rate in ICU patients with septic shock caused by CR-Kp and CR-Ab. Therefore, our preliminary data seem to encourage further clinical investigations on the role of NAC as an adjuvant therapy in ICU patients with septic shock due to multi-drug resistant Gram-negative bacilli.


Electromagnetic (EM) pollution has been evolving as one of the most concerning environmental problems in current society, due to the extensive application of EM technology, from household electronic apparatuses to wireless base stations, as well as military radars. Although both shielding and absorption are effective for resistance toward EM interference, the latter has received much more attention in recent years because it can convert EM energy and dissipate surplus EM waves . MicrowaConventional carbon black and graphite are rarely involved in carbon-based MAMs, because they easily suffer from impedance mismatching, and their relatively large size may induce serious phase separation in final composites. In contrast, carbon nanotubes (CNTs) and carbon nanofibers (CNFs) are more popular substrates for carbon-based MAMs, and their unique one-dimensional configuration has been verified to be favorable for accelerating electron transfer and consolidating conductivity loss . It is c3C2 nanosheets, and they found that the composite had much better microwave absorption performance than individual N-doped graphene or Ti3C2 nanosheets, whose minimum reflection loss and qualified absorption bandwidth reached up to −52.0 dB and 7.1 GHz, with an absorber thickness of 1.4 mm [The advent of graphene boosts the research of carbon-based MAMs significantly, and there have been over 4000 published papers on the fabrication of graphene-based MAMs, since its first report in 2011 . The typf 1.4 mm . It is oApart from these common members in the carbon family, carbon microspheres have also earned a place in the field of EM absorption. On one hand, their uniform spherical morphology makes it easy to disperse them into many solvents or resin matrix, and the serious agglomeration or re-stacking in CNTs/CNFs and graphene will not occur; on the other hand, the amorphous nature of carbon microspheres can create good impedance matching and bring more space for regulation of EM properties . There aSince the shape-preserving transformation of metal-organic frameworks (MOFs), it has rapidly developed into a dominant method for carbon-based MAMs. MOFs-derived, carbon-based materials usually possess some favorable features for EM absorption, including uniform morphology, porous microstructure, and well-dispersed metal nanoparticles. Although some significant achievements have been made in MOFs-derived carbon-based MAMs, a dilemma still hinders their popularization; that is, their composition and microstructure are in over-reliance on the corresponding MOFs precursors, and not in the optimum state for EM absorption. At present, composition optimization and microstructure design of MOFs-derived carbon-based MAMs are becoming a hotspot in the related field . Some adBiomass is considered as a kind of promising carbon source to construct carbon-based MAMs, not only for its abundant reserves and low cost, but more importantly for the fact that it ensures the formation of hierarchically porous microstructure in final carbon materials. In an effort to achieve better EM absorption performance, EM properties of biomass-derived carbon materials are generally regulated by introducing additional dielectric and magnetic components, through pre-impregnation or post-treatment methods . To date


A large university in Queensland, Australia with a diverse staff and student community introduced a campus wide smoke-free policy in 2016. The purpose of this enquiry was to understand attitudes about a new smoke-free policy, its potential impact and the shift in social norms and organizational culture to inform the next phase of implementation. An electronic survey was distributed to all staff and students approximately 12 weeks after the smoke-free policy was implemented. The survey consisted of multiple-choice questions about demographics, smoking behaviour, attitudes towards smoking and tobacco control, awareness of the smoke-free policy, and attitudes towards the effect of a completely smoke-free campus on quality of life, learning and enrolment. The survey was completed by 641 university staff and students. Respondents reported seeking out (80.4%) and socialising in smoke-free environments (86.6%) and supported smoke-free buildings (96.1%), indoor areas (91.6%), and outdoor areas (79%). The results revealed overwhelming support for a completely smoke-free campus (83%) and minority support for designated smoking areas (31%). Overall, respondents reflected positively towards a campus wide smoke-free policy. These findings suggest Queensland’s early adoption of tobacco control laws influenced the social environment, de-normalised smoking, changed behaviour, preference for smoke-free environments and shifted social norms. These findings provide convincing evidence for organisational change and suggest health promotion policy makers should progress the implementation of smoke-free policies nationally across the higher education sector. Tobacco smoking is the leading cause of preventable death and disease in Australia, contributing to more than 1 in 8 deaths or almost 21,000 lives in 2015 . HoweverIn the early 1980s, evidence emerged linking lung cancer in non-smokers with exposure to second-hand smoke . This evModifying the environment through the introduction of smoke-free legislation is an effective health promotion strategy for tobacco control. Sound evidence-based smoke-free policy underpinned by environmental change and supported by effective implementation, leads to behaviour change and a shift in social norms and organisational culture in settings such as university campuses. Australia has a long and successful history of smoke-free policies, with the first smoking restrictions commencing in the 1970s . AustralThe introduction of smoke-free policies has coincided with a reduction in reported respiratory problems, improved cardiovascular health and lung function among previously exposed non-smokers . These pStudies conducted in the United States confirm smoke-free policies positively impact smoking behaviour and exposure to second-hand smoke on university campuses ,15. FromBy 2016, twenty-two out of forty public universities in Australia had implemented smoke-free policies . UniversPrior to 1 July 2016, smoking was allowed in designated areas on campus in the University under investigation. From January 2016, the University communicated information about the new policy through several channels such as broadcast emails, the website, local forums and meetings and signage. The University allowed a twelve-month transition period. Transition strategies included advertising about implementation and operationalisation of the smoke free policy, marketing about smoking cessation resources including counselling, clear and regular communication signage, re-design of designated smoking areas, and staff training and education sessions. The new policy prohibits smoking in all areas of the university campus, including the grounds, buildings and vehicles. It includes the use of cigarettes and all other tobacco-related products including herbal cigarettes, loose tobacco, cigarette-making equipment and electronic cigarettes . The purpose of this study was to understand attitudes about the potential impact of a smoke-free policy and the shift in social norms and organizational culture to inform the next, institutionalisation phase at a large public university in Queensland, Australia.The setting for this study is a large public university in Queensland, Australia. The university has over 50,000 student enrolments—more than 38,000 are fulltime and nearly 9000 are international. The university employs over 4700 staff on two city campuses with a total revenue of $1.06 billion . StudentFollowing the implementation of the campus wide smoke-free policy in July 2016, an electronic survey was distributed to all current staff and students. An invitation to participate in this survey was included in the Registrars newsletter emailed during September 2016. The newsletter provided a short introduction about the research and a link to the survey. A follow-up email was sent in October 2016. The survey consisted of multiple-choice questions about demographics, smoking behaviour, attitudes towards smoking, tobacco control, awareness of the smoke-free policy, and attitudes towards a completely smoke-free campus on quality of life, learning and enrolment.n = 32) was used to measure reliability. The survey was anonymous and consisted of several multiple-choice questions. The survey used to collect data was developed and validated by researchers at a university in Western Australia . The queRespondents were grouped by their reported smoking status. Statement questions were developed to measure respondent’s attitudes towards smoking and the new smoking restrictions and indicated their level of agreement to statements using a five-point interval Likert-type scale, for example, strongly disagree/disagree/neutral/agree/strongly agree. Respondents were asked if they are aware of any [university] policy which restricts smoking on campus. A follow up question asked respondents to describe the university’s current smoking policy. Lastly, respondents were asked their opinion about the impact a smoke-free policy would have on staff and student quality of life, student learning, and student enrolment.The data were analysed using SPSS for Windows 10 and R . DescripOverall, 641 university staff and students completed the survey. The demographic representations of survey respondents and known demographics at QUT are described in n = 252, 45.8%) were exposed to second-hand smoke on campus. Of these, only 15.4% reported exposure to second-hand smoke once or more in a day, 25.5% reported exposure to second-hand smoke once or more than once a week, and 24.7% reported exposure to second-hand smoke a few times a month. A significant difference was observed in the level of exposure between current smokers, non-smokers and ex-smokers (p = 0.002). Less than half of the respondents  agreed with the statement “If someone smokes cigarettes around me, they are causing me harm”. A significant difference was observed in responses from current smokers, non-smokers and ex-smoker (p < 0.001), with 38.1% of current smokers agreeing with the statement. See Most respondents , “Rather date a non-smoker” , “Seek out smoke-free environments” , and were “Disappointed when a friend who does not normally smoke, smokes cigarettes while drinking alcohol” . In responses between current smokers, non-smokers and ex-smokers, significant differences were observed (p < 0.001) for all four statements. When asking others to not smoke around them, current smokers were less likely (3.2%) to agree with the statement (p < 0.001), compared to non-smokers (55.6%) or ex-smokers (25.8%).The overall attitude towards smoking was negative. Most respondents agreed they would “Prefer to socialise in a smoke free environment” , “All indoor worksites should be smoke-free, including bars and restaurants” , “The campus should be completely smoke-free”  and “The campus should be smoke-free including all outdoor areas” . Significant differences were observed between responses from current smokers, non-smokers and ex-smokers, (p < 0.001) for all statements, with current smokers less likely to agree than non-smokers or ex-smokers. See With reference to tobacco control attitude statements, most respondents agreed the “Campus should be smoke-free in all buildings” . Very few were unaware or “did not know” . No significant difference in responses between current smokers, non-smokers and ex-smokers was observed (p = 0.639). When asked to describe the current policy, most respondents affirmed “staff, students and visitors are banned from smoking tobacco throughout the campus; this includes all university buildings, grounds and vehicles” . No significant differences was observed in responses from current smokers, non-smokers and ex-smokers (p = 0.217).Regarding awareness about any [university] policy restrictions on smoking, most respondents knew about the new smoke-free policy  and students’ quality of life . Over half of respondents  felt the policy would have a positive effect on student learning. Less than half  felt the policy would have a positive impact on student enrolment. Significant differences were observed between responses from smokers, non-smokers and ex-smokers (p < 0.001) for all statements, with smokers less likely to acknowledge positive effects of the policy. See Most respondents agreed a smoke-free policy would have a positive effect on staff quality of life  agreed with a verbal reminder. Half  agreed with a monetary fine. Approximately one-third suggested anti-smoking education  or disciplinary processes for non-compliance . Less than a quarter  felt community service should be one of the consequences. Very few respondents  felt there should be no consequence for non-compliance with the policy.For successful implementation, respondents were asked about possible consequences for non-compliance with the smoke-free policy. Most respondents  compared to current smoking rates in Australia (12%)  and QueeThese results imply a compounding social, cultural and environmental effect stemming from the roll-out of smoking bans across Queensland for more than a decade and supports behaviour modification, social norming and organisational change through policy based environmental change. Especially relevant are policies modifying environments by prohibiting smoking at schools, restaurants, bars, workplaces, commercial eating and drinking areas and other public spaces such as public parks and beaches. This substantial shift in tobacco-related norms was confirmed by the majority preference towards a completely smoke-free environment (83%) and little support for preserving designated smoking areas (31%). These results correspond with the increasing denormalization and decreasing prevalence of smoking in Queensland. Many respondents agreed the policy would have a positive impact on students’ quality of life (84.5%), staff quality of life (82.1%) and student learning (63.5%). Overall, respondents reacted favourably to a complete smoking ban compared to those in Western Australia who responded less favourably in most questions . These fThe introduction of smoke-free policies in workplaces, restaurants and bars has been critical in reshaping the public perception of smoking, smoking behaviour and the demoralisation of tobacco in Queensland. In this instance, denormalization refers to the transition in the status of smoking from a socially acceptable and widely practised behaviour to one increasingly characterised as destructive, unhealthy and anti-social . ResearcIncreasing restrictions, about where smoking is permitted means people who smoke, are now being defined as a group whose behaviour entails segregation. Therefore, it is important for policymakers to consider how denormalization, social norms and organisational change affects those who continue to smoke within the context of policy-based environmental change. As smoking becomes increasingly concentrated among already disadvantaged populations , there is a risk these individuals and groups will feel even more marginalised and are less motivated to quit. Ongoing policy efforts to address the prevalence of tobacco smoking have reshaped the public perception of smoking as socially undesirable. This perception plays a major role in improving compliance with smoke-free policies by increasing feelings of dissonance or unease associated with nonconformity . This maYoung people in Queensland experience a smoke-free environment from childcare, go to smoke-free schools and later, they will work in smoke-free workplaces. University campuses and other tertiary settings are an obvious omission in the smoke-free educational facilities outlined The Tobacco Act and are not covered by current tobacco laws. Research confirms individuals who quit smoking before the age of thirty almost eliminate the risk of mortality due to smoking-induced causes . Thus, sAs more young adults attend university and the age of smoking uptake increases, campuses provide an important setting for primary and secondary smoking prevention and cessation support . These eFirstly, the response rate was low in this population. This might be because the initial survey was distributed during a mid-semester break, or the link was embedded in an electronic newsletter. Secondly, there is potential bias due to the small sample size. Thirdly, there was a large proportion of non-smokers, females and domestic students who participated in this study. This may be due to selective non-response and underreporting from current smokers, males and international students. To adjust or correct for this imbalance, results were post-stratified to reflect the QUT population according to gender, primary role and international student status. However, there may be an imbalance between other groups and/or a possibility for respondents to differ in their attitudes to non-respondents. These issues apply more generally to survey results and are difficult to control. The results from this study suggest support for a campus wide smoke-free policy. These findings may reflect the influence of Queensland’s early adoption of environment modification though the implementation of successive tobacco control laws and a consistent approach towards normalising smoke-free environments on attitudes and behaviour towards smoking, social norming and shifted organisational culture. The legacy of early adoption may continue to have a significant impact on the social acceptability and denormalization of smoking in public spaces. Comprehensive tobacco regulation at Queensland universities may deter young adults from initiating smoking and offer cessation support for those current smokers who are contemplating behaviour change.These findings have implications for the higher education sector because they contribute to the growing body of evidence for smoke-free policy implementation. These findings indicate a preference for smoke-free environments and a tolerance to smoke-free policies suggesting health promotion policy makers should progress the implementation of smoke-free policies nationally across the higher education sector.


Healthcare has undergone a significant transformation during the pandemic, with virtual services being rapidly developed and implemented to keep pace with societal needs. This study documented this change in healthcare by examining access and use of video-based, virtual service use before and during the pandemic. Participants for the study (n = 685) included three groups, including retirement-aged persons, middle-aged adults, and traditional college-aged students. Measures for the study included access to and utilization of physical and mental health services, satisfaction with services accessed, and anticipated access and use of virtual services following the pandemic. Results showed that most participants (94.2%) believed that virtual healthcare would persist after the pandemic; three-quarters of adults (75.2%) but only half of college-aged (52.8%) and retirement-aged (57.6%) participants anticipated using virtual healthcare in the future. Prior use and satisfaction with virtual healthcare services mediated anticipated future use for retirement-aged participants (p < .001), but only satisfaction with virtual healthcare was a marginal predictor for college-aged participants (p = .051), and neither were predictors for adult-aged participants. These results support that people believe virtual healthcare will persist after the end of the pandemic, but that there are age-related differences in who anticipates using these services in the future, and which factors will make the most difference in attracting clients. These differences can impact how healthcare providers market and develop further tele-health services to increase the likelihood of use by retirement-aged participants, and suggests that client satisfaction is a key mediator for different age groups.


Single-cell sequencing technologies have led to a revolution in our knowledge of the diversity of cell types, connections between biological levels of organization, and relationships between genotype and phenotype. These advances have mainly come from using model organisms; however, using single-cell sequencing in non-model organisms could enable investigations of questions inaccessible with typical model organisms. This primer describes a general workflow for single-cell sequencing studies and considerations for using non-model organisms . Importantly, single-cell sequencing, when further applied in non-model organisms, will allow for a deeper understanding of the mechanisms between genotype and phenotype and the basis for biological variation. Cells are the fundamental units of life, varying in form and function within and between individuals, populations, and species. The sequencing of nucleic acids within single-cells, known as single-cell sequencing (scSeq), has led to a revolution in our knowledge of the diversity of cell types at varying levels of the biological hierarchy , while also advancing our understanding of genotype x phenotype relationships. As in most of biology, our knowledge of humans and model organisms  far exceeds that of non-model or wild species, which remains a barrier to resolving how organisms function in nature. At present, this remains the case for scSeq; although scSeq is a promising and proven method, its use has been limited and its potential applications in non-model systems have yet to be realized. Nevertheless, there have been a few examples where scSeq has already been used in non-model organisms to investigate questions that are inaccessible with typical model organisms .The fast pace of technological advancement in the field, requirement of organism-specific technical knowledge, and cost are all barriers to applying scSeq to non-model organisms. The first challenge for applications of scSeq in non-model organisms is its recency; since the first study published in 2009 . AnotherThis primer describes a general workflow for scSeq studies and includes considerations for using non-model organisms. Although many genomic characteristics can be investigated with scSeq , we focus on single-cell DNA sequencing (scDNA-seq) for variant calling and single-cell RNA sequencing (scRNA-seq) for expression quantification as these two approaches are foundational technologies. Importantly, researchers should also consider single-cell multiomic approaches, where multiple genomic features are investigated in the same cell . FinallyThe specific aims of a proposed study will determine the amount and type of background knowledge necessary , the specific protocols , and data analysis techniques used. Indeed, scSeq projects have been performed on diverse organisms such as lizards , ants 66, fish , spongesResearchers should assess the availability and quality of the focal organism’s reference genome before embarking on a scSeq project, as many protocols rely on reference-based methods . For speAs scSeq has become more popular, sample collection protocols and library construction have become robust and highly optimized. Today, preparing single-cell suspensions remains a key challenge faced by researchers who wish to apply this approach in non-model species . With anThe first strategy, whole-cell sequencing, is very similar to protocols used in human studies, where fresh samples are collected and processed to prepare single-cell suspensions . To prepOnce tissues arrive in the lab, generating single-cell suspensions and library construction are both time-sensitive steps. A challenge for specimens from non-model organisms is deciding the optimal tissue dissociation methods, as organism-specific protocols do not exist in most cases. Generally, all dissociation protocols consist of mechanical digestion and enzymatic digestion . In the Single-nucleus sequencing is an alternate strategy to achieve comparable results as whole-cell sequencing . Single-2+ that serve to stabilize the integrity of nuclei [For single-nucleus sequencing, the integrity of nuclei is critical because nucleic acids may escape disassociated nuclei, causing failure or bias in downstream sequencing. Nuclei can be extracted using either the rapid, efficient and practical  method of nuclei . The visDNA isolated from a single cell is the target unit for scDNA-seq . The majWhole-genome amplification attempts to overcome this limitation of insufficient genomic starting materials by amplifying the genome. However, amplification methods can be biased and mutagenic due to PCR errors, thus affecting downstream interpretation of genomic variation .Single cell multiple displacement amplification (SCMDA) is a technique that amplifies the genome without using sequence-specific primers and improves the annealing of hexamer primers . These cLinear Amplification via Transposon Insertion (LIANTI) is a technique that randomly fragments the genomes of single cells using transposases . The DNASingle-stranded sequencing using microfluidic reactors (SISSOR) uses microfluidics to amplify long segments of separated DNA strands, resulting in the sequencing of complementary and homologous DNA strands . SNPs arThe Linked-Reads Assay from 10X Genomics was a popular approach that used droplet microfluidics for scDNA-seq. This product provided long read sequences that could be used to phase haplotypes and identify structural variants (10X Genomics protocol CG00044). This product was replaced by a newer technology, 10X Genomics Chromium Single Cell CNV, which used a different cell capture workflow (10X Genomics protocol CG000246). 10X Genomics discontinued their genomic products due to a legal injunction, brought about by BioRad’s case of patent infringement . However, this injunction was vacated, clearing a path for 10X Genomics to resume sale of the Chromium Single Cell CNV and Linked-Reads products . However, at present, 10X Genomics does not have a scDNA-seq solution, so there are no high-throughput, commercially available scDNA-seq solutions.Like scDNA-seq, scRNA-seq is limited by the small amount of starting material. Messenger RNA isolated from a single cell is the target unit for single cell transcriptomics. Different levels of mRNA abundance are used to identify differentially expressed genes, and to assign cell types. There are many methods for scRNA-seq, and the methods vary in the type of cell isolation required, sensitivity, and amplification biases. Broadly, different high-throughput scRNA-seq workflows can be divided into how the single cells are separated, often based on the question being answered: droplet-based and microwell strategies.Droplet-based strategies, where droplets containing single cells are produced, is a broadly available technology. Drop-seq and InDrop are two approaches that require cells to be isolated through droplet microfluidics ,36. ThesChromium Single Cell Gene Expression from 10X Genomics is a widely used, commercially available platform that also uses droplet-based microfluidics. Chromium uses a technology, “Gel bead in Emulsion” (GEM), which encapsulates single cells and reagents within the microfluidic chip. RNA extraction, reverse transcription, and barcoding occur in the GEMs . CurrentMicrowell strategies, where microfluidics is used to partition single cells into microwells, is an alternative approach. Seq-well is one of the newer protocols using microwells, whereas this method has been employed in previous forms through Smart-seq, CEL-seq, MARS-seq, and mcSCRB-seq are four microwell-based approaches that require cells isolated onto microfluidic plates ,41,42. SWhile scSeq enables transcriptomic profiling of thousands of cells, the position of these cells within a tissue is lost after sample preparation. Since the debut of spatial transcriptomics in 2017, the field has been pushed further by adding spatial transcriptomics to study the cellular transcriptional atlas without losing tissue context. There are several methods to perform spatial transcriptomics. Popular methods can be grouped into two approaches: (1) using in situ capture, which relies on barcoded bead arrays to capture mRNA molecules with oligo tails ; (2) using fluorescence in situ hybridization such as Slide-seq. Each method has their own advantage such as the 10X visium can provide faster and more robust results while Slide-seq can provide a higher resolution. Because spatial transcriptomics cannot provide transcriptome profiling at single-cell resolution as each capture spot represents an area with ~10 cells depending on the tissue type, computational methods have been developed to combine scRNA-seq and spatial transcriptomics to compensate for the loss of resolution in spatial transcriptomics ,45,46. AUnique considerations are required when analyzing scSeq data due to the nature of sparse data dropout events and the potential for stochastic gene expression. Genetically identical cells may show different levels of gene expression caused by the nature of gene expression being determined through a Poisson process. These challenges are exacerbated for non-model organisms without high-quality reference genomes. These unique challenges add to variation among both technical and biological replicates. In the case of scRNA-seq, it is particularly challenging to distinguish technical variation (transcripts are not detected in some samples) and biological variation (differences in transcript abundances among samples). For single-cell DNA sequencing, the major limitation is data dropout events due to low-coverage sequencing and amplification biases. For example, parsing technical variation due to amplification bias apart from low-expressed transcripts or rare alleles may result in inflation of cell-to-cell variability.Depending on the data analysis method, preprocessing of data may be required before further computational analysis . Two impDepending on the question being answered, after preprocessing scRNA-seq data, several analytical and statistical methods can be applied. Two common goals with single-cell RNA data are cell-level analysis and gene-level analysis . With ceA typical scSeq analysis includes the following steps: (1) raw data pre-processing, where raw sequencing reads are demultiplexed and then filtered based on unique feature counts, total unique molecules detected and percentage of reads mapped to mitochondria; (2) data normalization, such as global log transformation of the gene expression; (3) linear dimensional reduction based on highly variable features from cell-to-cell, where this step determines the dimensionality of the dataset; (4) non-linear dimensional reduction that uses previously determined clusters to visualize and explore the dataset; and (5) identification of differentially expressed features and cell type assignment which is based on the expression of biomarkers of each cell type. Additional analyses including trajectory prediction and integration of different omics also can be applied.Well-developed software suites for analyzing scSeq data such as Seurat ,46 and SFor cross-species single-cell studies, the workflow can be extended based on the identification of cells of interest using cell-type-specific biomarkers from different species ,55. By dDosage compensation describes the biological phenomenon that produces equivalent expression between a gene located on the X or Z sex chromosome and its ancestral autosomal gene . Equal gMeiotic sex chromosome inactivation (MSCI) refers to the transcriptional silencing of genes located on sex chromosomes within germ cells . MSCI apLivestock research can benefit greatly from scSeq, largely through improving our understanding of reproductive traits. A unique characteristic of reproductive traits is that the performance is fundamentally reflected by only a few cells, such as different stages of oocytes, and supporting cells such as granulosa cells. As there are many factors to determine the performance of reproduction at different developmental stages, traditional research methodology such as bulk RNA-sequencing may introduce noise from supporting cells. Normal procedures to quantify gene expression usually require steps to remove or isolate target cells such as oocytes. For example, the primary oocyte is more difficult to isolate from the secondary oocyte . BesidesAn important feature of scSeq is to identify cell types without isolating and purifying target cells. scSeq also provides an opportunity to study interactions between cell types. For example, it can be used to study the oocytes and supporting cells that play a crucial role shaping reproduction. scSeq, especially scRNA-seq, has been widely applied to human reproduction research. It has been extensively used to study the complex gene expression network in multiple cell types such as primordial germ cells ,66. For Overall, these examples illustrate how scSeq can be applied in non-model organisms help us expand our understanding of the genetic or functional basis of traits, which have relevance for foundational knowledge as well as for practical applications that benefit humanity.Although there are difficulties associated with scSeq for non-model organisms, we argue that this is the direction the field must go in to accurately infer biological processes from genomic variation and to build a more mechanistic link between genotype and phenotype. Importantly, work with non-model organisms requires substantial evaluation of pre-existing resources, such as the availability and quality of a reference genome, protocols to isolate and digest cell types of interest, and compatibility with bioinformatic workflows. Non-model organisms may be the most suitable for answering specific basic and applied questions. Overall, scSeq technologies, when applied to non-model organisms, may reveal unique biological processes and further our understanding of the link between genotype and phenotype.Looking into the future we can think about the eventual destination for scSeq technologies for non-model organisms. For example, in model organisms including humans, major initiatives are underway to develop cell atlases for tissues, organs, and the whole body through developmental stages, for example the Tabula Sapiens . Similar


Our aim is to determine the strong predictors of the onset of instrumental activities of daily living (IADL) decline in community-dwelling older people.A prospective cohort study with a two-year follow-up.Kashiwa City, Chiba Prefecture, Japan and Toshima Ward, Tokyo Metropolitan, Japan.The data were acquired from two cohorts. The final sample comprised 1,523 community-dwelling older people aged 65–94 years . They were individuals who were independent in IADL at baseline and participated in follow-up IADL assessments two years later.At baseline, comprehensive assessments were performed including: health interview, gait function, hand-grip strength, skeletal muscle mass, balance function, oral function, dietary lifestyle, cognitive function, quality of life, mental status, and social network. When the two-year follow-up was performed, IADL declines were observed in 53 out of 1,523 people. The association of each Z-transformed parameter with the occurrence of IADL decline was examined by employing a binominal logistic regression model adjusting for age, gender, body weight, body height, and medical history. An odds ratio (OR) and a 95% confidence interval were calculated and compared between different parameters.A decrease in walking speed and one-legged stance time, whereas an increased timed up & go test time was associated with significant ORs for the occurrence of IADL decline.Gait-related parameters appear to be the strong predictors of the onset of IADL decline in community-dwelling older people. Maintaining sufficient mental and physical capacity is the key to successful aging. According to Lawton’s model, functional capacity can be divided into the following seven levels of competence: 1) life maintenance; 2) functional health; 3) perception-cognition; 4) physical self-maintenance; 5) instrumental self-maintenance; 6) effectance; and 7) social role [There are several indicators which may serve as predictors of the onset of IADL dependence in community-dwelling older people –13. For et al. proposed that in order to identify older people who need aid, a short-term predictive value of the indicators would be more substantial than a long-term predictive value [As described above, the predictors of the onset of IADL dependence in the older people has been well studied –13. Howeve value . Then, ave value . TherefoThe Toshima cohort study was approved by the ethics committee of Tokyo Metropolitan Institute of Gerontology , while the Kashiwa study was approved by the ethics committee of the University of Tokyo (number 12–8). This ethics approval is in accordance with the Declaration of Helsinki (1964). All the participants in this study provided written informed consent.The data of two different cohorts in Japan was utilized. The first cohort was observed in the Kashiwa study which took place from 2012 to 2014. The second cohort was observed in the Toshima study from 2014 to 2016. Both studies were conducted to investigate age related functional changes including physical, cognitive and social functions in community-dwelling older population in Japan. The original papers have already reported studies on the Kashiwa  and ToshThe Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC) was employed to evaluate the IADL status in participants. TMIG-IC was developed to assess the following three high-level of functional capacities of older people in Japan: (1) IADL, (2) Intellectual activity, and (3) Social role. The IADL list of TMIG-IC consisted of five questions . The parThe medical history of the participants in regard to chronic diseases such as hypertension, stroke, diabetes mellitus, hyperlipemia, chronic renal failure, heart disease, and cancer was recorded by nurses.2), and appendicular skeletal mass index (ASMI) (kg/m2) were measured as physical structure measurements. ASMI (kg/m2) was estimated by using a bioelectrical impedance analysis system .Body height (cm), body weight (kg), body mass index (BMI) , one-legged stance time (s), usual walking speed (m/s), maximum walking speed (m/s), and timed up & go (TUG) time (s) ,20. A deOral function was assessed by a speech diadochokinetic test  . For theet al. (2003) and has been used to assess the dietary habits of older people in Japan [Dietary habits were evaluated by using the Dietary Variety Score (DVS). This index was developed by Kumagai in Japan . The parDepressive symptomatology was evaluated using the Japanese version of the Geriatric Depression Scale (GDS) . This asThe WHO-Five Well-being Index (WHO-5) was used to measure the current subjective Quality of Life (QOL) which assesses positive mood, vitality, and general interest . The tesA Mini-Mental State Examination (MMSE) was performed to evaluate the cognitive functioning among the older people . The scoThe Lubben Social Network Scale (LSNS-6) was used to measure social engagement including that of family and friends . This teP < 0.05 as the significance level. All statistical analyses were performed using the SPSS software .A Pearson chi-square test and a Mann-Whitney U test were employed to compare baseline characteristics between the IADL decline and IADL non-decline groups. The Mann-Whitney U-test was performed to compare the baseline parameters between the dropout (n = 796) and non-dropout (n = 1523) groups. Neither those who dropped out nor those who did not have any impairment in IADL at baseline. Binomial logistic regression analyses (by the forced entry method) were performed using either “with IADL decline” or “without IADL decline” as the dependent variable. Each parameter served as the predictor variable. These analyses were then adjusted according to baseline age, gender, body weight, body height, and medical history of illnesses . The statistical results were shown as an odds ratio (OR) with a 95% confidence interval (CI). To compare the results of each predictor variable, a Z-transformation was applied to each parameter prior to the binominal logistic regression analysis. A Hosmer-Lemeshow test was used to assess the fit of the logistic regression model with P < 0.05). Additionally, the Pearson chi-square test showed a significant difference in percentage of gender and percentage of history of cancer between the participants of IADL decline and IADL non-decline groups (P < 0.05).P > 0.05). Four of the variables  were significantly associated with the occurrence of IADL decline (P < 0.05), and showed larger ORs than the other variables.P = 0.028; one-legged stance time, P < 0.001; maximum walking speed, P < 0.001; TUG time, P < 0.001).The Mann-Whitney U-test was performed to compare the baseline parameters between the dropout (n = 796) and non-dropout (n = 1523) groups. There was no significant difference in age between the dropouts and non-dropouts. The dropout group had lower motor function than the non-dropout group  in Japan. The TMIG-IC related to IADL status includes more complex daily activities such as transportation, shopping, meal preparation and money management compared to the more basic daily activities [Previous studies have shown that many variables are associated with physical function, cognitive function, and social role. These are often identified as the predictors of future IADL decline in the older people ,9,12,13.tivities . The abitivities . MaintaiThis study found that parameters related to gait were important for predicting the occurrence of IADL impairments. Although we did not measure gait patterns, biomechanical and neurophysiological studies have shown age-related changes in gait patterns –32. TherSeveral studies reported that a decrease in cognitive function is related to the decline of BADL and IADL ,11. CognThe previous study shows that the social role performance assessed by TMIC-IG is significantly associated with future IADL decline . Lawton’Of late, oral functioning has attracted attention from researchers and practitioners. Previous studies show that poor oral health is significantly associated with physical frailty, sarcopenia, and mortality among the community-dwelling older people . HoweverThis study had several limitations. 1) The present data were obtained from Japanese suburban areas ; thus, the present results may differ in rural areas as well as other countries. 2) The proportion of participants with IADL decline was small; hence, the participants with IADL decline may have dropped out of the follow-up assessment. The motor function of the dropouts was lower than those who did not drop out. Although a power analysis was performed to confirm that the number of samples was large enough, generalization of the results needs careful consideration. Indicators of relatively low significance  may become significant when more representative samples are included. Thus, cognitive, social, and oral functions could be used as secondary predictors of future IADL decline in the older people. 3) A comprehensive assessment was performed in this study, which concluded that gait-related parameters were the strongest predictors of future IADL decline. However, the unexamined measures, such as socio-demographic factors, may provide results different from those of the present study. 4) The final sample size was very small compared with the total number of participants recruited. Although the participants were randomly selected, sample selection bias may have occurred in this study.In conclusion, this study revealed that gait-related parameters such as TUG time, maximum walking speed, and usual walking speed are significantly associated with future IADL decline in community-dwelling older people. Daily monitoring of these indicators associated with gait-function would be useful for detecting future risk of IADL decline.