Obstructive sleep apnea (OSA) is a common and costly medical condition. Untreated OSA is associated with numerous and well-documented adverse health consequences including depression, diabetes, cardiovascular disease, and premature death. In addition to these health consequences, untreated OSA is also associated with substantial costs borne by patients, payers, the health system, and society at large. Perhaps more importantly, evidence suggests that OSA treatment is associated with positive economic benefit. The purpose of this brief review is to introduce economic aspects of OSA, including the potential economic benefit of OSA treatment. Approximately 14% of men and 5% of women between the ages of 30 and 70 years in the United States suffer moderate to severe OSA10, with 936 million individuals estimated to suffer OSA worldwide11. In part because of rising rates of obesity, the prevalence of the disorder is increasing13.Obstructive sleep apnea (OSA) is a common and costly medical condition. The health consequences of OSA include increased risk for cardiovascular disease14. Yet, despite widespread acknowledgment that OSA is associated with substantial economic burden, relatively little research has evaluated economic aspects of OSA treatments15. In the modern healthcare climate of rising costs on the one hand and limited resources on the other, such knowledge is essential for payers and policy-makers who seek to make evidence-based decisions regarding allocation of scarce healthcare resources. To this end, the purpose of the present paper is to introduce key health economic concepts pertaining to OSA and to very briefly summarize the evidence to date regarding health economic aspects of OSA.Beyond well-documented adverse health outcomes, OSA incurs dramatic economic costs borne by patients, payers, employers, and society16. Broadly speaking, these OSA costs can be divided into three categories: direct costs, indirect costs, and health-related quality of life (HrQOL). Direct costs of OSA include costs of OSA diagnosis and management, including provider consultation, sleep apnea testing, device and supply costs , prescription medications, and so on. Indirect costs include costs unrelated to direct OSA care that are nonetheless attributable to OSA, such as increased healthcare utilization (HCU) for non-OSA conditions, diminished workplace productivity , and increased risk of accidents and errors including motor vehicle collisions (MVCs). In the AASM white paper, the greatest costs associated with OSA were lost workplace productivity ($86.9 billion), increased HCU ($30 billion), MVCs ($26.2 billion), and workplace accidents and injuries ($6.5 billion)16. Although total societal-level costs are often cited to demonstrate the costs associated with OSA and other sleep disorders, it is important to realize that each of these costs is primarily borne by different stakeholders17.Several years ago, the American Academy of Sleep Medicine commissioned a white paper that estimated total societal-level costs of OSA to exceed $150 billion per year in the United States alone19. HrQOL includes both general as well as disease-specific outcomes, which can be measured using various tools. Common general HrQOL measures include the PROMIS, developed as part of the National Institutes of Health Roadmap20, Short-Form 36 (SF-36)21, EuroQol-5D22, and others; the Functional Outcomes of Sleep Questionnaire (FOSQ)23 is an example of an OSA-specific measure of HrQOL. The most common economic metric to quantify HrQOL is the quality adjusted life year (QALY). QALYs represent the product of HrQOL and time  and thus provide researchers and decision-makers with a standardized metric to compare return-on-investment across diverse disease states, such as in cost-effectiveness analyses (CEAs)15. In CEAs, cost-effectiveness is determined based on the ratio of quality to cost, which is known as the incremental cost-effectiveness ratio (ICER). As a measure of value, ICERs reflect the cost per QALY. A given health system or payer determines a willingness to pay (WTP) for a QALY. For example, in the US, a cost of $50,000 per QALY is a generally accepted threshold for cost-effectiveness. Over the past three-plus decades, a preponderance of evidence demonstrates that untreated OSA worsens HrQOL and OSA treatments improve HrQOL and are generally cost-effective14.In addition to direct and indirect costs that can be clearly monetized, HrQOL is a key economic outcomeThe sleep medicine ecosystem includes diverse stakeholders such as patients, payers, providers, health systems, and equipment manufacturers, just to name a few. Each of these stakeholders incurs different costs  so perceives the value of sleep differently. For example, patients desire improved quality of life, such as increased energy for work or leisure activities, as well as ease of care experience. Payers want to reduce costs of care delivery, which is directly related to profits. Providers want valid, reliable, and easy-to-administer testing and treatment options, and so on. 24. Kim and colleagues found that payer costs were reduced by $264 over three months when patients were treated within the at-home arm (as opposed to the in-lab arm). By contrast, provider operating margin was reduced from $142 to –$161 under these same conditions. Finally, time horizon is an important consideration: most OSA treatment costs are front-loaded, but OSA cost reductions might not be realized immediately.From an economic perspective, the diagnosis of OSA involves two broad components: provider time and sleep apnea testing. As a result, the most expensive costs would include a face-to-face physician encounter, in regulated hospital space, and in-lab diagnostic polysomnography with an additional in-lab CPAP titration. At the other extreme, diagnostic costs could include consultation with a mid-level provider, via telehealth, and home sleep apnea testing (HSAT) without a separate titration study. Like OSA diagnosis, OSA treatment costs include two components: initial device and equipment (e.g. CPAP) and ongoing supply costs. Furthermore, it is important to realize the competing interests in these OSA pathways: cost savings (a positive) from the payer’s perspective results in reduced revenue (a negative) from the health system’s perspective or decreased reimbursement (a negative) from the provider’s perspective. As one example, consider findings from an economic analysis of a multi-site clinical trial designed to compare in-lab versus at-home approaches to OSA diagnosis and treatment17.As healthcare transitions from volume-based care to value-based care, alternative payment models (APMs) have been employed in numerous disease states. To date, APMs have not yet impacted sleep medicine in the US in a meaningful way. Nonetheless, bundled payments and shared risk are likely to impact the field of sleep medicine sooner rather than later. Within a value-based framework, both maximizing quality and minimizing cost are essential. Such matters, including cost-shifting between stakeholders, have been discussed in detail elsewhere25 or included only a very limited number (n = 5) of studies that examined the impact of OSA treatment on HCU26. More recently, our group published a systematic review of the impact of OSA treatments on monetized economic outcomes. Results demonstrated that 15 of 17 comparisons reported a positive economic benefit from OSA treatment 41, which might have contributed to the positive findings. Similarly, Chhatre and colleagues analyzed data from a large national Medicare administrative claims database32. When the authors used CPAP machine charges as a proxy for objective CPAP use, CPAP was associated with reduced total costs. Interestingly, de Vries and colleagues33 found OA therapy more cost-effective than CPAP in terms of QALYs, but not in terms of reduction in apnea-hypopnea index, over 12 months. Results of these and other studies examining the economic impact of OSA treatments, reflecting a non-systematic update since our prior systematic review Relative to thousands of studies that evaluated pathophysiologic, clinical, and epidemiologic aspects of OSA, few studies have considered the economic impact of OSA treatments. Two prior reviews primarily considered costs of untreated OSA42. Evidence suggests that such racial sleep disparities might underlie racial disparities in chronic disease outcomes, such as cardiovascular disease and diabetes43. Relative to whites, blacks are also less likely to adhere to CPAP therapy44, and low socioeconomic status (SES) and neighborhood of residence have also been associated with poorer CPAP adherence46. These results further support a link between OSA disparities and health disparities. In recognition of the importance of sleep disparities in overall health disparities, the National Institutes of Health recently hosted a workshop and identified research needs; a summary report has been published elsewhere and is recommended for review47.Racial and socioeconomic factors have been identified as an important determinant of OSA and OSA outcomes. For example, relative to whites, blacks suffer higher rates of OSA but are less likely to be diagnosed48. Third, it is essential to adopt the employer’s perspective when assessing the economic impact of OSA. It is notable that the majority of OSA-related costs are borne by employers, yet very few studies have considered economic aspects of OSA from the employer’s perspective. Fourth, given the positive economic impact of CPAP adherence, much greater insight is needed regarding the costs and effectiveness of interventions to improve CPAP adherence49. Finally, given the very rapid technological advancements in OSA diagnosis and treatment, it will be important to evaluate the economic impact of telehealth and remote monitoring for OSA. This recommendation is especially important given the impact of the COVID-19 pandemic in terms of clinical care and rapid adoption of telehealth approaches. Additional actionable recommendations to advance understanding regarding the health economic aspects of OSA are presented in Our most important recommendation is to include economic outcomes in OSA clinical trials and sleep research more broadly. Public–private partnerships are likely to be especially fruitful in this regard. Second, in light of the high rates of comorbidity of OSA with numerous medical and psychiatric conditions , it will be important to evaluate economic aspects of OSA within various disease subpopulations. Disentangling the effects of OSA relative to the often better studied comorbid conditions, such as cardiovascular disease, is methodologically challenging yet vitalIn summary, OSA incurs significant economic costs borne by a wide range of stakeholders including patients, payers, health systems, industry partners, and others. Although important questions remain unanswered, a small but considerable body of evidence suggests that the diagnosis and treatment of OSA is associated with positive economic benefit. Our most important recommendations are for researchers to include economic endpoints in all OSA clinical trials and to adopt the economic perspectives of multiple stakeholders in the sleep medicine ecosystem. In the modern healthcare climate of rising costs on one hand and limited resources on the other, economic aspects of care will become increasingly important determinants of health policy and resource allocation.


Paralympic Powerlifting is a sport in which the strength of the upper limbs is assessed through bench press performance in an adapted specific bench. It is therefore essential to optimize training methods to maximize this performance. The aim of the present study was to compare force production and muscle activation involved in partial vs. full range of motion (ROM) training in Paralympic Powerlifting. Twelve male athletes of elite national level in Paralympic Powerlifting participated in the study . The athletes performed five sets of 5RM (repetition maximum), either with 90% of 1RM in full ROM or with a load of 130% 1RM in partial ROM. All subjects underwent both exercise conditions in consecutive weeks. Order assignment in the first week was random and counterbalanced. Fatigue index (FI), Maximum Isometric Force (MIF), Time to MIF (Time) and rate of force development (RFD) were determined by a force sensor. Muscle thickness was obtained using ultrasound images. All measures were taken pre- and post-training. Additionally, electromyographic signal (EMG) was evaluated in the last set of each exercise condition. Post-exercise fatigue was higher with full ROM as well as loss of MIF. Full ROM also induced greater. EMG showed greater activation of the Clavicular portion and Sternal portion of pectoralis major muscle and lower in the anterior portion of deltoid muscle when full ROM was performed. Muscle thickness of the pectoralis major muscle increased post-exercise. We concluded that training with partial ROM enables higher workloads with lower loss of muscle function. Paralympic Powerlifting (PP) is a paralympic sport that focus in maximum strength development and the best athletes are able to lift three times their body mass . The parAs a strength modality, Paralympic Powerlifting training includes control of several variables, such as load, strength, movement duration and bar displacement velocity , 3. BesiWeightlifters and others involved in resistance training commonly perform exercises with partial ROM to displace superior amounts of load or even to complete a target number of repetitions during a set. However, there is a lack in the literature regarding different ROM during weightlifting performed by Paralympic athletes.Therefore, the aim of the present study was to compare the fatigue index, the maximum isometric force, the time to maximum isometric force, the rate of strength development, the muscle thickness and the activation of the muscles involved in partial vs. full range of motion (ROM) in Paralympic Powerlifting.The participants consisted of 12 male Paralympic Powerlifting athletes with 28.60 ± 7.60 years, 71.80 ± 17.90 kg and with a minimum training experience of 12 months. To enable seated weighing, weigh-in was performed on a digital electronic platform-type Michetti  with a maximum supported weight of 3000 kg and a size of 1.50 x 1.50m. All participants were national-level competitors who met the necessary prerequisites of the Brazilian Paralympic Committee to be eligible for the sport . Five atThe study lasted for three weeks. In the first week the subjects were familiarized with measurement of electromyography, ultrasound and mechanical strength assessment procedures; and were also submitted to 1RM testing and re-testing. An interval of 48-h was kept between each session of testing on week 1.On weeks 2 and 3 the individuals were submitted to two different exercise conditions in a counter balanced design. So, half of the athletes underwent a training session with full ROM and the other half underwent training with partial ROM. For the partial ROM, four wood boards 2.5 cm thick were used, as shown in Before each intervention, the athletes performed a 10.min warm-up for the upper limbs comprising three sets of 10 to 20RM in three exercises  , 12. TheThe 1RM test was performed in the week of familiarization (week 1), where each subject chose the load they believed could be lifted using the maximum effort. Weight increments were then added until the maximum load was reached. If the subject could not perform a single repetition, 2.5% of the load used in the test was subtracted . The subThe subjects were submitted to the tests of FI, MIF, Time, RFD, thickness and muscular activation. Following this, after 10 minutes of rest , the athThe subjects were submitted to the tests of FI, MIF, Time, RFD, thickness and muscular activation. After 10 minutes of rest , they unMeasurement of muscle strength, fatigue index (FI), Maximum Isometric Force (MIF), Time to MIF (Time) and Rate of Force Development (RFD) were determined by a force sensor PFMA 3010 , fixed to the adapted bench press bench using Spider HMS Simond carabiners  with a rupture load of 21 KN. A steel chain with a breaking load of 2,300 kg was used to secure the force sensor to the seat . PerpendMaximum Isometric Force was measured by the maximum force generated by the upper limb muscles. The MIF was determined by the isometric force, measured by the force sensor and the adapted bench press bench, which was adjusted so that there was an elbow angle close to 90°, and at a distance of 15 cm from the starting point (chest to bar). Angle of the elbows was verified with a goniometer FL6010 . Participants were instructed to perform a single maximum movement seeking elbow extension (as soon as possible) and, after movement, to relax for MIF evaluation. For the fatigue index (FI) assessment, the same exercise was performed and the subjects were asked to maintain the maximum contraction for 10-s. FI was determined by FI = -1,0)X100). The rate of force development (RFD) was measured through the relationship between time and force development (RFD = Δforce/Δtime), adapted from the adapted methodology of Milner-Brown, Mellenthin and Miller [® SSD 500V ultrasound (US) with a 7.5 MHz electronic linear transducer . Ultrasound images were captured using a 90 dB gain and a magnification that allows a depth of 42.0 mm of the Pectoralis major muscle. Images were acquired between the Sternal and Clavicular portion. Participants remained relaxed for 10 min to allow fluid distribution before and after training [Muscle thickness (MT) was obtained using an Alokatraining . Particitraining , 18. Thetraining , 20.Maximum isometric force measurement and electromyographic signal were evaluated simultaneously. Surface electromyography (EMG) is a noninvasive technique with no contraindications, which assesses the action potential of muscle fiber activity over which the sensors are placed .EMG analysis used EMG System EMG 432C muscle equipment. EMG targeted the Pectoralis major sternal portion (LSP) and clavicular portion (CP), the anterior deltoid (AD), and the long head triceps brachii (LHTB), which were captured during maximal voluntary 10-s isometric contractions (MVIC). There was an elbow angulation of approximately 90°, with a distance of 15.0 cm from the bar to the stern . The surp2. When significant interactions were identified, post-hoc pairwise analysis was performed along with Cohen´s d for effect size. EMG records post-training with the exercise conditions were compared with paired t-test. The level of statistical significance was set at p ≤ 0.05. Results are presented as means (X) ± standard deviation (SD).The statistical analysis was performed using the Statistical Package for Social Science, version 22.0. Normality and sphericity assumptions were confirmed with Shapiro Wilk and Mauchly tests, respectively and were then followed by a 2x2 repeated measures ANOVA, which was performed on the following dependent variables: Rate of force development, maximum isometric force, fatigue index, time to maximum isometric force and muscle thickness in clavicular and sternal portions of pectoralis major. The factors (interaction terms) were the exercise condition  and the moments in time of assessment (pre- vs. post-training). In the 2x2 ANOVA, the effect size was given by the ηp2 = 0,720). The same was observed for time to maximum isometric force .In the remaining variables significant interaction of exercise condition vs moment in time were detected and therefore pairwise comparisons between moments in time were investigated for each exercise conditions separately. Maximal isometric force decreased in the full Rom condition , and also, though with a smaller effect, in partial ROM . The fatigue index increased significantly solely in the partial Rom condition , but did not in full ROM . The clavicular portion of the pectoralis major muscle thickness increased from pre- to post in both exercise conditions, with greater effect in full ROM , compared with partial ROM . Finally, the sternal portion of the pectoralis major muscle thickness also increased from pre- to post in full ROM  and in partial ROM p<0.001; d = 2.36).The aim of the present study was to compare the fatigue index, the maximum isometric force, the time to maximum isometric force, the rate of strength development, the muscle thickness during training with partial vs. full range of motion (ROM) in Paralympic Powerlifting.The main results of the present study were that training with full ROM induced greater muscle signs of fatigue (greater decrease in maximum isometric force and greater increase in fatigue index).Training with full ROM seems to induce a greater muscle fatigue, as shown in the present study. Moreover, training with partial ROM enables higher training loads. Indeed, Sullivan et al., , have obRegarding the edema measured through ultrasonography, it was found that the muscle thickness changed significantly from pre- to post-test exercise in both training conditions. Changes were greater when the subjects performed the full ROM training  and a significant interaction between time of measurement and exercise conditions was evident. Indeed, post-exercise muscle thickness was larger in the full ROM condition, despite a lower load being lifted.Our study partially corroborates that by Pinto et al., , who useThe fact that the subjects were able to move a higher load (130% 1-RM) in partial ROM with a smaller muscle edema, seems to favor this training strategy when maximum strength is the main goal. Indeed, the body of knowledge on recovery mechanisms post-inflammation supports the rationale that larger edema requires enlarged recovery time .Previous studies on bench press exercise  and on eIn the present study, muscle activation was different between the two exercise conditions. During full ROM pectoralis major muscle was the most active muscle group, whereas in the partial ROM it was triceps and deltoid muscles. On the other hand, Lander et al.,  found thThe main limitation of the present study was the small number of participants. The specificity of this populations does not enable large scale studies and therefore we must rely on the accumulation of more contributions before a clear tendency is established. Another limitation is that the differences herein do not necessarily support possible differences in adaptations to partial ROM training as apposite to full ROM training.Training through partial range of motion has been used in Paralympic Powerlifting. However, to the present, possible differences and / or similarities between this method and that with full range of motion were not clear. Post-exercise fatigue indicators were higher with full ROM . We conclude that training with partial ROM enables handling of higher workloads with lower loss of muscle function.S1 Dataset(XLSX)Click here for additional data file.


Dear Editor:uninfected. Regarding conjunctival swabs, all tested eyes yielded virus-negative results. Clearly, this is important empirical information in regard to infection risks for ophthalmologists and medical staff in tertiary eye care centers, including the handling of human corneal donor tissue — always provided that patients or donors are not infected.In a prospective study, Rokohl and colleagues looked into the possibility that SARS-CoV-2 may hide in the eyes of pre-screened asymptomatic patients in a tertiary eye care center . The autinfected individuals, viz., COVID-19 patients, survivors, or asymptomatic carriers, and virus material persistence in immune-privileged sites of the eye and elsewhere?We compliment the authors for taking this initiative and call for research regarding hiding places of SARS-CoV-2 to be expanded: What about infected individuals may be(come) of interest, in 1967 a virus reservoir in sperm was proposed in the course of discovering the Marburg virus (MV) [To exemplify why “secret” hiding sites in rus (MV) . Carefulrus (MV) ) spermatrus (MV) . That Ebrus (MV)  — in semrus (MV)  — conveyRegarding SARS-CoV-2, investigating virus materials in semen of COVID-19 cases in acute and recovery phases yielded ambiguous results. The question of whether virus material may persist in the privileged immunity site of the testes remains open . In cont


Virus Mutations, Integration sites and Cis-effects (ViMIC), which specializes in three features: virus mutation sites, viral integration sites and target genes. In total, the ViMIC provides information on 31 712 VMs entries, 105 624 VISs, 16 310 viral target genes and 1 110 015 virus sequences of eight viruses in 77 human diseases obtained from the public domain. Furthermore, in ViMIC users are allowed to explore the cis-effects of virus-host interactions by surveying 78 histone modifications, binding of 1358 transcription regulators and chromatin accessibility on these VISs. We believe ViMIC will become a valuable resource for the virus research community. The database is available at http://bmtongji.cn/ViMIC/index.php.Molecular mechanisms of virus-related diseases involve multiple factors, including viral mutation accumulation and integration of a viral genome into the host DNA. With increasing attention being paid to virus-mediated pathogenesis and the development of many useful technologies to identify virus mutations (VMs) and viral integration sites (VISs), much research on these topics is available in PubMed. However, knowledge of VMs and VISs is widely scattered in numerous published papers which lack standardization, integration and curation. To address these challenges, we built a pilot database of human disease-related  Meanwhile, millions of virus sequence data deposited in public databases may be used to discover feature patterns related to diseases. Domain knowledge that can be used to improve the understanding of virus cis-effects, for instance, gene expression and transcription cis-regulatory information, has been deposited in freely accessible databases, such as Cistrome Data Browser ,19 and tVirus Mutations, Integration sites and cis-effects (ViMIC). ViMIC is built to fill such gaps from the aspects of virus mutation, integration, and cis-effect of host. The unique features of our database over other databases include: ViMIC is the first database to provide VMs and virus sequence data annotations retrieved from a large collection of public domains; and ViMIC is the first to visualize VMs harbored in VIS fragments and integrate VIS data with cistrome resources to help researchers who are interested in gene regulation mechanisms to systematically investigate the functional annotation of VISs in the host genome. ViMIC mainly covers three special features: virus mutation sites, viral integration sites and target genes. Firstly, the database facilitates the search for VM annotations and quick browsing for statistics on virus sequences. Secondly, the database can help users determine whether the integration site of viral fragments harbor mutations or whether the VIS integrated into the host genome is in a functional element distinguished by histone modification, transcription factor binding or chromatin accessibility. Thirdly, the database provides information on host target genes affected by viral insertion or by virus gene/protein/region, and on the correlation between gene expression levels and the fraction of infiltrating immune cells in human diseases.Therefore, to overcome the challenges mentioned above, we present a database of human disease-related A schematic overview of the ViMIC data collection, text-mining system, data processing and web interface is summarized in Figure The major sources of data in ViMIC were derived from the published literature, the Cistrome Data Browser ,19, VISDThe Entrez Programming Utilities (E-Utilities) tool was used to build a set of full names and abbreviations of virus keywords into a fixed URL syntax to search and retrieve virus-related articles from PubMed. A Python script was run to filter the articles by matching keywords (e.g. mutation) as regular expressions. Full-text articles from PMC and abstracts from PubMed (BioC-XML format) with annotations of biomedical concepts in the Bioconepts2pubtatorcentral file were downloaded from PubTatorCentral , and thiFor the virus sequence module, GenBank id, virus gene, genotype, geographical region and disease information was extracted from the NCBI Nucleotide database for standardized annotation and statistics. For the VIS-VM module, virus integration fragments were collected from publicly available literature. Since the information of some collected VISs may be incomplete, we only focused on those VISs where both breakpoint locations and the viral reference genome were provided. The multiple sequence alignment tool CLUSTALW  was thentrans-acting factor on a genome-wide scale, also known as the in vivo genome-wide location of transcription factor binding-sites or histone modifications’. Therefore, the host gene expression may be influenced if the virus integrates into the Cistrome. Cistrome DB collects human and mouse ChIP-seq, DNase-seq and ATAC-seq data from publicly available resources. In our current study, Cistrome DB data was applied as cistrome factor information to integrate with data from a virus integration site and assist users to explore the cis-effect of VIS. The procedure included four steps:To construct features for exploring the functional integration of a VIS (VIS-factors), the overlaps were calculated between the VIS and cistromes. The concept of ‘cistrome’ has been widely used in the gene regulation field ,19,35. AFirstly, the viral integration sites in the human genome were derived from VISDB (76 264 curated VISs) and from the PubMed literature  that reported disease-related viral integration events in the human genome. The collected VISs were processed into BED format, which was then converted into human reference genome coordinates in GRCh38 by the LiftOver if a previous genome version was used. Secondly, the ChIP-seq peaks of human transcription factor and histone modifications were downloaded as well as the DNase-seq and ATAC-seq peaks from the Cistrome Data Browser. This information was then compressed into bgzipped (.gz) files . Thirdly, a GIGGLE (genome search engine)   (1.6 (40 bp) for all VISs with two positions, which indicated that two-point VIS with length <40 bp would have similar overlap calculation result with the single site. Therefore, for each VIS with two points, the overlapping calculation results of single start site and two positions will be both displayed. Moreover, the classification case of VISs with two positions based on literature (case 1 or case 2) and the length (the distance of two positions) were also added in the web interface for those VIS with two positions.The location used in ViMIC is according to the coordinate in the human genome of VIS declared in the original articles. There are two forms of VIS location, one is VIS with a single site, and the other is VIS with both two positions. In our data, the single site accounts for nearly 94%. VISs with two positions were generally classified into two categories. Case 1: the deletion of human fragments during the integration event  ; Case 2: et al.) ,39. We c et al.) , and vis et al.) . SupplemViMIC is characterized by a comprehensive curation of VMs, functional annotations of VISs in the host genome, and target genes. ViMIC provides three features, as shown in Figure The ‘Virus Mutation Site’ feature provides two modules for users: the VM annotation module and the Sequence module. The VM annotation module includes mutation site, mutation level, virus gene/protein/region, virus-associated disease, genotype/subtype, and literature evidence. A description column indicates more detailed information for each mutation site using the following tags: (i) D: drugs mentioned in the literature; (ii) G: genotypes/subtypes mentioned in the literature; (iii) R: reference genomes mentioned in the literature; (iv) S: viral sequences mentioned in the literature; (v) I: the VM is related to immune escape; (vi) Cl: clinical information available in the literature and (vii) C: geographical distribution of the source mentioned in the literature. The virus sequence module provides the curated information and virus sequence data statistics including a map of viral sequence source distribution and statistics of virus sequence in the viral gene, genotype and countries. Adenovirus-associated virus type 2 (AAV2) was excluded in this feature because there is no pathogenic mutation information from current studies.The second feature is the VIS-cistrome factor overlap calculation, which was specifically designed to explore whether the VIS is involved in a functional region in the host genome. For each VIS, if there are overlaps between the VIS and the genomic peaks of the cistrome factors (sum of overlaps > 0), the detailed results are presented, including the overlap number and annotation of each cistrome sample . Furthermore, all the VIS-cistrome factor overlapped records on human chromosomes are summarised using heatmap and histogram plots, and the top-ranked cistrome factors ordered by the overlap number in each chromosome using stacked bar plots. Thus, users can view the overlap distribution between the VIS and cistrome factors in the human genome. In the current version of the VIS-VM module, VISs of three viruses including hepatitis B virus (HBV), human papillomavirus virus (HPV) and AAV2, with precise breakpoint coordinates and the reference genome in the integration literature, were used to map and visualize whether mutation occurred in VISs.This feature provides information for the target genes affected by viral insertion or by the virus gene/protein/region. We have systematically annotated the target gene information, including official gene symbols, official full names, gene aliases, transcripts, gene types, gene location information, gene functions, gene IDs and drug information. For each gene, if there were available gene expression data for patients with the virus-related disease, we further analysed the relationship between its expression and immune cell infiltration and represented it by heatmap and lollipop plots.In the current version as of 21 May 2021), the database contains 8 viruses, including HBV, HPV, Epstein-Barr virus (EBV), AAV2, Merkel cell polyomavirus (MCV), human immunodeficiency virus (HIV), human T-cell lymphotropic virus type-1 (HTLV1) and xenotropic murine leukaemia virus-related virus (XMRV), as well as 31 712 VMs entries, 105 624 VISs  entries, 17 778 436 overlapping entries between VISs and Cistromes, 77 virus-associated diseases , 16 310 target genes, 1 110 015 virus sequence information records and 127 datasets with clinical information. The detailed statistics for ViMIC data are listed in Table  May 2021All of the annotation and analysis results are deposited and managed in a MySQL relational database on a Linux server. More detailed information on the VMs, VISs, and target genes is crosslinked to the related resources including PubMed, VISDB, ENCODE, NCBI GenBank , WikipedViMIC allows users to search data of interest in three ways. Firstly, a quick search is available on the homepage. Users can input a keyword of interest including a virus name, disease, histone mark, transcription regulator or virus gene/protein/region. Secondly, there is a search engine at the top-right corner of each table in the web interface, where users can enter any keywords of interest. Thirdly, an advanced search is also available on the home page for sophisticated searches by selecting a virus name of interest, and searching by human chromosome, disease, virus gene/protein/region or target gene. Each search produces tables of matched entries. Users can then view the detailed information by clicking on the hyperlink or the ‘View’ button in the table.A web interface was designed to provide user-friendly access to browse, search, analyse, visualize and download data. The database provides an interface for each virus with a summary and links to three ViMIC features to help users quickly check their hypothesis regarding VMs, VIS-cistrome factor overlaps or viral target genes.Firstly, the ‘Virus Mutation Site’ feature presents a catalogue of links to virus genes proteins/ regions and diseases on the homepage of selected virus. In the ‘Mutation’ module, users can view a particular VM labelled with literature PMID, virus gene/protein/region and description. Moreover, users can click the ‘Sequence’ button to browse and download the curated sequence annotation of the selected virus. The detailed virus mutation page is composed of mutation annotations and evidence from the literature.Secondly, the ‘Viral Integration Site’ feature provides the sum of VIS-cistrome factor counts and three VIS-cistrome factors overlap counts in each human chromosome, as well as two browse data menus for users to view overlapped entries by chromosomes or factors on the homepage of selected virus. Users can browse VIS information by clicking the ‘View’ button in the table as well as by selecting a chromosome or cistrome factor in the browse data menu. ViMIC also developed a function on the web page that allows users to upload a list of integration sites and query them in our database, or users can upload their own VISs location information to the ViMIC and will be notified by email once the job is completed. Moreover, ViMIC designs a VIS-VM module of selected virus to visualize mutations observed in VIS fragments and a search module to show overlap data according to options of a user's interest including disease, biological source . In addition, ViMIC provides a heatmap of three VIS-cistrome factor overlap counts on each chromosome and two stacked barplots of top-ranked cistrome factors to help users have a better understanding of the distribution of VIS-cistrome factor overlap in the human genome.Third, ‘Target Gene’ feature provides the curated target genes affected by viral genome insertion or by virus gene/region/protein. Users can browse the detailed gene information, related VISs/VMs and the correlation analysis results between gene expression levels and the proportions of 22 types of infiltrating immune cells in virus associated human diseases.ViMIC also includes virus, disease, dataset, download and help modules on the homepage, which allows users to be quickly redirected to a particular feature of users’ interest. In addition, users can browse datasets for which clinical characteristics and virus sequencing data are available in the literature in our dataset module. To make the data presentation more intuitive, the statistical plots and tables are also shown on the home page of each module.In this article, we have taken HBV as an example, as it is a viral infection that can cause death in patients, even in anti-viral drug users.Users can click the green button with VM number of HBV in the virus mutation overview page, as shown in Figure et al. identified the recurrent HBV insertion nearby the telomerase reverse transcriptase (TERT) gene  gene . In ViMIRT) gene . In HepGRT) gene . TherefoRT) gene . All of As TERT is a target gene of ‘1000606’, ViMIC also presents the immune analysis between TERT gene expression and 22 types of infiltrating immune cells based on CIBERSORT Figure . The detet al. studied HBV interactions with human chromatins and investigated the significance of four histone modifications  on HBV integration segment . GIGGLE can search and rank the overlapping genomic loci between query genomic locations such as VISs and genome interval files and it is available as a GitHub repository at https://github.com/ryanlayer/giggle. LiftOver is a web service that provides genome coordinate transformation between different human genome reference assemblies and can be freely accessed at https://genome.ucsc.edu/cgi-bin/hgLiftOver. PubTator is a web service that can automatically annotate biomedical entries including genes and mutations, from the literature, which is available at https://www.ncbi.nlm.nih.gov/research/pubtator/. CIBERSORT uses the gene expression data to estimate the abundances of member cell types from a mixed cell population, and it can be freely accessed at https://cibersort.stanford.edu/.ViMIC is available at gkab779_Supplemental_FileClick here for additional data file.


Text CorrectionThere was an error in the original article . The worDimerization of the MoMuLV genome occurs co-transcriptionally or near the site of the provirus [53–57] whereas HIV genomes dimerize randomly , most likely after being exported from the nucleus [59] and possibly not until they reach the plasma membrane [30].The authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected. The original article has been updated.References Added54J. Virol. 2003, 77, 4577–4587.Onafuwa, A.; An, W.; Robson, N.D.; Telesnitsky, A. Human immunodeficiency virus Type 1 genetic recombination is more frequent than that of Moloney Murine Leukemia Virus despite similar template switching rates. 55J. Virol.2004, 78, 12129–12139.Flynn, J.A.; King, S.R.; Telesnitsky, A. Nonrandom dimerization of murine leukemia virus genomic RNAs. 56Retrovirology2005, 2, doi:10.1186/1742-4690-2-3.Kharytonchyk, S.A.; Kireyeva, A.I.; Osipovich, A.B.; Fomin, I.K. Evidence for preferential copackaging of Moloney murine leukemia virus genomic RNAs transcribed in the same chromosomal site. 57J. Gen. Virol.2006, 87, 2279–2289.Rasmussen, S.V.; Pedersen, F.S. Co-localization of gammaretroviral RNAs at their transcription site favours co-packaging. 58J. Virol.2003, 77, 11193–11200.Rhodes, T.; Wargo, H.; Hu, W.-S. High rates of human immunodeficiency virus type 1 recombination: Near-random segregation of markers one kilobase apart in one round of viral replication.


Virus infection is one of the threats to the health of organisms, and finding suitable antiviral agents is one of the main tasks of current researchers. Metal ions participate in multiple key reaction stages of organisms and maintain the important homeostasis of organisms. The application of synthetic metal-based nanomaterials as an antiviral therapy is a promising new research direction. Based on the application of synthetic metal-based nanomaterials in antiviral therapy, we summarize the research progress of metal-based nanomaterials in recent years. This review analyzes the three inhibition pathways of metal nanomaterials as antiviral therapeutic materials against viral infections, including direct inactivation, inhibition of virus adsorption and entry, and intracellular virus suppression; it further classifies and summarizes them according to their inhibition mechanisms. In addition, the use of metal nanomaterials as antiviral drug carriers and vaccine adjuvants is summarized. The analysis clarifies the antiviral mechanism of metal nanomaterials and broadens the application in the field of antiviral therapy. Virus infection has always been a threat to human and animal health. Typical viruses include hepatitis B virus , influenAs one of the emerging fields in recent years, nanomaterials can be simply divided into one-dimensional, two-dimensional, and three-dimensional nanostructures according to the morphology of the material. There are many methods to synthesize nanomaterials, such as mechanical grinding synthesis , chemicaThis review first provides a more comprehensive summary of the different stages of the role of metal nanomaterials in inhibiting viral infections in recent years. (1) Directly inactivating the virus; (2) inhibiting virus adsorption and entry; (3) intracellular virus suppression. Subsequently, it summarizes the use of metal-based nanomaterials as drugs and vaccine carriers. The application of metal nanomaterials in the field of antiviral research is introduced and its prospects are discussed. This review focuses on metal nanomaterials by comparing research progress in related fields in recent decades ,20,21. TVirus infection of cells can be divided into three stages. The first is the early adsorption of the virus onto cell membrane, where the virus binds to cell surface receptors through its surface protein. The second is the process of uncoating, replication, and translation after the virus enters the cell. Finally, the virus repackages to release new progeny viruses. In these processes, if metal nanomaterials can inhibit the virus in the early stage and prevent the virus from invading cells, they can be used in medical protective equipment, which has great potential for preventing the virus spread .2 nanomaterials with photocatalytic properties can generate active substances by using light or through their affinity with biological macromolecules. They can damage the biofilm and kill the bacteria. Since the surface of the virus also has a biofilm, these metal-based nanomaterials can be used for the direct inactivation of the virus. H. Cui and colleagues found that TiO2 has good photocatalytic properties and can generate reactive oxygen species (ROS) under ultraviolet irradiation. ROS has oxidative properties. Therefore, using external interference in tumor therapy, the balance of ROS expression in tumor cells will be affected, which accelerates tumor cell apoptosis. At the same time, its excessive presence can also destroy the biofilm and cause it to lose its biological activity. Related genes in cells regulate the normal expression level of ROS. H. Cui’s group explored the inhibitory effect of TiO2 on the influenza virus under ultraviolet irradiation  on the cell membrane . In thisAu/Ag is known to have a good affinity with S. Jinyoung Kim synthesized Au and Ag with different morphologies and with or without holes. After co-cultivating with the virus for a certain period, the adsorbed and unadsorbed viruses were separated by centrifugation. The interaction between the nanoparticles and the virus was observed by transmission electron microscope, and the number of disulfide bonds in the viral hemagglutinin was quantified to determine the effective destruction of the disulfide bonds by the metal-based nanoparticles. The results show that, compared with non-porous Au and Ag nanomaterials, porous Au nanoparticles can achieve a more apparent antiviral effect by destroying the disulfide bonds in the hemagglutinin on the surface of the virus . This prThe process of a virus invading cells involves specific binding, which provides a good starting point for preventing viruses from infecting cells. The primary inhibition method is to competitively bind to the virus by simulating cell surface virus receptors and inhibiting the virus from invading cells. However, this method can stop the virus from spreading further and does not inactivate the virus. There are certain drawbacks to the application. It is worth noting that these nanomaterials can be adsorbed on medical protective fabrics and concrete surfaces, thereby reducing the spread and infection of viruses.After the virus enters the cell, it uses the intracellular machinery to carry out protein replication and translation. Since the virus is uncoated to expose the genetic material DNA/RNA, this process can provide the possibility for external drugs to destroy the viral nucleic acid or interfere with the process of translation, replication, and release.2S nanoparticles to explore their inhibitory effect on coronaviruses. The author explored the expression levels of the virus by adding Ag2S materials at different times after virus infection. Since the particle size of Ag2S is about 2.5 nm, it has a more profound cell penetration ability and can better interact with viruses. PEDV nucleocapsid (N) protein is a protein that binds to viral RNA, and its expression is tested to verify the inhibition of Ag2S. The results show that Ag2S inhibits the virus by inhibiting the negative-strand RNA synthesis and budding of PEDV [When the virus enters the cell, it begins the process of unpacking, replication, and translation. During this period, the RNA/DNA and protein of the virus are synthesized in large quantities; this happens to be the most effective moment for drugs to inactivate viruses. By importing the drug into the cell, it acts on the viral RNA/DNA and protein, destroying the replication process of the virus and inhibiting its reproduction. Most nanomaterials have good biocompatibility. The advantages of small particle size and high specific surface area can help nanomaterials enter cells efficiently. With the capability of binding metal ions with biological macromolecules, the metal nanomaterials electrostatically interact with viral nucleic acids or proteins to form chemical bonds, destroy their structures, or cause irreversible conformational changes of viral proteins, thereby achieving the purpose of inhibiting virus replication. Ting Du synthesized glutathione-encapsulated Ag of PEDV .The relatively high surface activity of metal nanomaterials also brings the disadvantage of high cytotoxicity. However, more amphiphilic polymers such as PEG and PVP have been studied at this stage . They caThe high infectivity and pathogenicity of viruses has caused people to pay more attention to antiviral drug research and development. The current research and development of viral drugs mainly include screening natural drugs and synthesizing organic small-molecule drugs. However, the screening of natural medicines requires a high workload, and the inhibitory effect of natural medicines on viruses is not satisfactory. The inhibition mechanism remains to be explored. The research and development of organic small-molecule drugs, finding the relevant derivatives of existing antiviral small-molecule drugs, and further exploring drugs with viral inhibitory effects have good application prospects. However, small organic molecules have disadvantages such as low water solubility, poor biocompatibility, and high toxicity. Moreover, the development and application of vaccines have also opened up new areas for virus therapy. Vaccines are preventive treatments that can significantly reduce the incidence of viral infections. However, vaccines for injection have biological activity, and instability in transportation and storage is still an open question. Hence, many scholars have developed numerous carrier materials, such as colloids , magnetiThe human body contains many kinds of metal elements, including Al, Ca, Mg, Fe, Na, Mn, Zn, K, Li, Cu, Se, etc. These metals are essential for participating in various life cycle processes in the human body. Therefore, small amounts of metal ions are less harmful to the human body, and the metal ions could be prepared as nanomaterials with a high specific surface area of nanomaterials, porosity of some materials, and good biological activity as a promising drug carrier.The poor water solubility of antiviral drugs makes their bioavailability low, and higher doses are often required to achieve the desired therapeutic effect. However, higher doses will produce certain toxicity to organisms, so the emergence of drug carriers can improve the drugs’ bioavailability and reduce the damage to organisms. When selecting metal nanoparticles as drug carriers, the toxicity of metal ions to organisms and the loading efficiency of materials should be considered. Using metal ions or inert ions, with higher content in human cells, can reduce the toxicity of materials and increase the materials’ natural metabolism.Human Acquired Immune Deficiency Syndrome (AIDS), caused by HIV, is a disease in which immune cells are the primary targets. These cells can accumulate viruses for a long time to be used as targeted sites for drug delivery. Studies have shown that Au nanoparticles can be more internalized by human macrophages and have less biological toxicity . HinojalQuantum dots (QD) are a type of low-dimensional semiconductor material and often have smaller sizes. Therefore, applying QD materials to the field of biological therapy can often improve the therapeutic effect. Ranjeet Dungdung’s group used ZnS quantum dots as a drug carrier, loaded with mycophenolic acid (MPA), an immunosuppressant against dengue fever virus. It was found that cells have a higher internalization rate of ZnS-coupled MPA, and its inhibitory effect on dengue virus was significantly improved and the selectivity index was increased by two orders of magnitude . The stu2O4@BaTiO3, AZTTP is bonded to nanomaterials’ surface through ionic bonds. As shown in The application of antiviral drugs can significantly reduce the infection rate of the virus. However, the presence of the blood–brain barrier reduces the scope of action of the drug, which is also a shortcoming in the therapeutic application of most drugs. When Madhavan Nair and colleagues studied the antiretroviral application of HIV, they found that the antiretroviral drug azidothymidine 5′-triphosphate (AZTTP) has low efficiency in passing through the blood–brain barrier, which makes the treatment of the virus not very efficient. Therefore, it is crucial to find an excellent carrier to load AZTTP to achieve targeted drug release. Studies have shown that by synthesizing 30 nm magnetic CoFeIn the treatment of viral infections, in addition to traditional antiviral drugs, nucleic acid drugs such as oligonucleotides , siDNA , DNAzymeStudies have shown that small interfering RNA 331 (siRNA331) can target the conserved region of hepatitis C virus RNA . InspireVaccines are used to prevent infectious diseases by artificially attenuating and inactivating pathogenic microorganisms and their metabolites to produce an immune response in the body. After immunization, the organism will produce immune antibodies to protect itself, and the immune system will form a memory of pathogenic microorganisms. When it invades again, the immune system can react in time and produce corresponding antibodies to protect the organism. The treatment of viral infections is often more complicated. However, it is often a valid preventive method to cause the body to produce specific antibodies through the previous vaccine injection.Vaccines can be divided into live-attenuated vaccines, inactivated vaccines, recombinant vaccines, and so on. Among them, live vaccines  content in the cell is higher than that in the extracellular tissue, the synthetic material undergoes a redox reaction between GSH and the disulfide bond in the material to release OVA. Studies have shown that the material’s better transportation performance increases the immune effect and the memory of the immune system. Furthermore, metal elemental nanomaterials have a small particle size and are easily taken up by cells. Therefore, the cellular uptake of antigens and adjuvants that they carried increases [ZIF-8 has mild synthesis conditions, and it is possible to encode ovalbumin (OVA), an antigen model, into ZIF-8 for co-transport of antigen and adjuvant by surface electrostatic adsorption of immune adjuvant cytosine-phosphate-guanine (CpG) . Accordincreases ,66. Morencreases . Moreovencreases ,69.Due to the excellent physical and chemical properties of metal-based nanoparticles, their biological applications have been continuously expanded. Previous work has found that some metal-based materials themselves have adjuvant properties. During the early vaccination process, alum is a promising adjuvant widely used in vaccines. Later studies found that CAP can stimulate the organism’s immune response, and its effect is better than that of alum . On this2+ to prevent protonation in organisms and improve the stability of antigens [Some adjuvants need to be added into inactivated virus vaccines to increase the immune effect when used in vivo. Here, adjuvants can stimulate the immune response, and protect the activity of the inactivated virus antigen. The inactivated foot-and-mouth-disease virus vaccine is not stable when used. The histidine residues on the surface protein are protonated, causing it to lose immune activity ,72. Metaantigens . Similarantigens . In termantigens .We briefly summarize below the therapeutic mechanism of nanomaterials in viral infection inhibition. As shown in The spread of viruses such as SARS-CoV and SARS-CoV-2 as well as influenza viruses poses a huge threat to human health. It is generally known that viruses are prone to mutate due to external influences to produce different virus subtypes, limiting the use of traditional antiviral drugs and vaccines. Therefore, novel drug discovery and vaccine development for the treatment of viral infectious diseases is very important.2, etc., have virus-inactivating ability and can damage their surface proteins or their genetic material by combining with viruses. In comparison, nanoparticles such as ZnO and Fe3O4 can prevent the virus from further infecting cells by interfering with the replication, translation, and release of the virus. Some other metal nanoparticles have a small particle size and good biocompatibility, are easily taken up by cells, and can be used as a suitable carrier for antiviral drugs. Moreover, utilizing the binding of metal ions and biological macromolecules, metal nanoparticles can be used as vaccine adjuvants or adjuvant carriers to promote the occurrence of immune response of the body. However, there are still some problems to be solved in the antiviral application of metal-based nanomaterials. First of all, the toxicity of metal ions in organisms is still a major obstacle to their application. The biological toxicity of metal ions has always been a major obstacle to the application of metal-related materials in biological treatments. While ensuring their therapeutic effect, reducing the in vivo toxicity of metal ions is one of the most important directions for researchers to explore. At this stage, there are mainly two methods; first, by reducing the concentration of metal materials used, and second, by optimizing the metabolism of metal ions. However, the effect is not yet satisfactory. Next, the antiviral mechanism of some metal ions has not yet been explored clearly. Finally, there are many metal nanomaterials, and the current research is limited to a few metal ions. The antiviral properties of other metals still need to be further studied. Based on their excellent properties, the application of metal-based nanomaterials in the antiviral field is a promising research field for expansion.Metal-based nanomaterials have the advantages of high specific surface area and small particle size, making them widely used in the biological field. In terms of antiviral activity, some gold-based nanomaterials such as Au, Ag, CuI, TiO


The emergence and re-emergence of infectious diseases are a cause for worldwide concern. The introduction of Zika and Chikungunya diseases in the Americas has exposed unforeseen medical and logistical challenges for public health systems. Moreover, the lack of preventive measures and vaccination against known and emerging mosquito-transmitted pathogens, and the occurrence of unanticipated clinical complications, has had an enormous social and economic impact on the affected populations. In this study, we aimed to measure the seroprevalence of endemic and emerging viral pathogens in military personnel stationed in Manaus, Amazonas state.We measured the seropositivity of antibodies against 19 endemic and emerging viruses in a healthy military personnel group using a hemagglutination inhibition assay (HIA).Flaviviruses varied depending on the virus serogroup. Additionally, 0.6% and 0.3% individuals were seropositive for Oropouche virus (OROV) and Catu virus (CATUV) from the family Peribunyaviridae, respectively. All samples were negative for Eastern Equine Encephalitis virus (EEEV), Western Equine Encephalomyelitis virus (WEEV), Mayaro virus (MAYV), Mucambo virus (MUCV) and CHIKV from the family Togaviridae.Overall, DENV positivity was 60.4%, and 30.9% of the individuals reacted against ZIKV. Also, 46.6%, 54.7%, 51.3% and 48.7% individuals reacted against West Nile virus (WNV), Saint Louis encephalitis virus (SLEV), Ilheus virus (ILHV) and Rocio virus (ROCV), respectively. Individuals with high DENV HIA titer reacted more frequently with ZIKV or WNV compared to those with low HIA titers. Observed cross-reactivity between A high proportion of individuals in our high-risk population (~ 60%) lacked antibodies against major endemic and emerging viruses, which makes them susceptible for further infections. Military personnel serving in the Amazon region could serve as sentinels to strengthen global infectious disease surveillance, particularly in remote areas.The online version contains supplementary material available at 10.1186/s13071-021-04901-4. Deforestation, urbanization and climate change have led to perpetual public health challenges of infectious diseases worldwide . In the Dengue virus (DENV), one of the foremost studied arboviruses, is still responsible for millions of infections and numerous deaths worldwide . ConsecuThe social and economic impacts attributed to the recent Zika virus (ZIKV) and Chikungunya virus (CHIKV) introductions, driven by the large sheer number of human infections and associated pathology in Brazil and the Americas, are well documented , 11. In Furthermore, compared to civilian population, military personnel live in a communal nature, train in diverse locations like the Amazon rainforest and participate in humanitarian aid in adverse conditions, alongside suboptimal hygiene and stress in the field, which increases their risk of contracting emerging infectious diseases. Hence, soldiers can act as a sentinel population to identify emerging pathogens. However, we have few data about the serological status before or during recent arbovirus outbreaks and the role of cross-reactive antibodies against these emerging viruses. Hence, in this study we evaluated the prevalence and antibody reactivity among military personnel participating in the jungle survival course using a cell-based assay against major endemic and emerging arboviruses from three different virus families. Additionally, we performed a literature review to understand the distribution of DENV prevalence in Brazil between 1980 and 2020.Seroprevalence greatly varies depending on the study population, age, sex and serological assay employed for antibody testing. Based on previous estimates of seroprevalence for arboviruses in Brazil, which ranged between < 1% to > 50% depending on the geographical location , 19–22, The study population comprised of Brazilian army personnel participating in a jungle survival course at the Jungle Warfare Training Center , located in Manaus, Amazonas State. Every year CIGS organizes up to four training camps in the Amazon rainforest, where recruits spend a maximum of 3 months inside the rainforest, training and performing exercises. We interviewed and sought ethical consent from participants before entering the rainforest and collected blood samples after the end of the jungle survival course. Number of participants varied with each training camp. Adults of both sexes ≥ 18 years were invited to participate. During the study period there were no female participants. Consecutive individuals were enrolled in this observational and cross-sectional study using convenience sampling until attaining calculated sample size of 300 between January 2014 and December 2015. A total of 4 ml of blood was drawn from each participant , subsequently tubes were centrifuged and plasma was separated and stored at −80 °C until further analysis.Flaviviridae (Flavivirus genus): yellow fever virus (YFV), dengue virus (DENV) serotypes 1 to 4 , Zika virus (ZIKV), Saint Louis encephalitis virus (SLEV), West Nile virus (WNV), Ilheus virus (ILHV), Rocio virus (ROCV); Togaviridae (Alphavirus genus): Eastern equine encephalitis virus (EEEV), Western equine encephalomyelitis virus (WEEV), Mayaro virus (MAYV), Mucambo virus (MUCV), Chikungunya virus (CHIKV); Peribunyaviridae (Orthobunyavirus genus): Oropouche virus (OROV), Tacaiuma virus (TCMV) and Catu virus (CATUV).The serological tests were performed at the Instituto Evandro Chagas (IEC) . Plasma samples collected were subjected to an in-house hemagglutination inhibition assay (HIA) with a titration cut-off of 1:20 plasma dilution, as previously described , 24. Samhttps://www.saude.gov.br/boletins-epidemiologicos).PubMed, Science Direct, LICACS, Web of Science and Medline databases were searched by keywords Fig.  to ident2 of the inverse titer value, calculated with the formula normalized_titer = Log2[1/titer]  . We thenFlaviviridae family (genus Flavivirus) . Our entire sample was negative for all five viruses tested from the Togaviridae family , B (n = 59) and C (n = 169)  and B (n = 2).We then evaluated the clustering of individuals according to their serological titers against all viruses tested. We observed that our population clustered into three groups: A (69) Fig. . Group AFlaviviruses, we analyzed the reactivity of DENV-1- or DENV-2-positive individuals with their corresponding reactivity for DENV-2 or DENV-1, WNV or ZIKV .Furthermore, 5% (15/295) and 17.2% (51/296) individuals self-reported previous malaria and dengue virus infection, respectively. We performed a chi-square test to assess whether self-reported dengue infection rates were similar to results observed by serological diagnosis performed in this study. We observed that serological testing showed a higher percentage of positive individuals than self-reported disease  strain 17D compared to the wild-type strain. Moreover, the decreasing reactivity toward wild-type strain may be an indicator of a gradual decrease of antibody titers over time after vaccination. YFV booster doses at 10 years after primary vaccination in travelers to endemic regions, endemic populations and high-risk individuals such as military personnel are necessary to heighten the 17DD-YF-specific immune response and to achieve efficient immunity .Flaviviruses tested in this study. We cannot rule out multiple flavivirus infections in the same individual. These findings, along with other reports, indicate that SLEV, WNV, ILHV and ROCV circulation in Brazil is largely unknown, and there may be epidemiological implications of the co-circulation of these arboviruses [Flavivirus infections in cross-protection or induction of a severe disease [Flavivirus infections.The presence of heterologous neutralizing antibody titers is inversely correlated with the severity of patients with a second DENV infection , 33–35. oviruses , 19. Ove disease , 15, 36. disease –39. Rece disease , 41. On  disease , 43. NevAlphavirus tested in our study. Similarly, we observed very low prevalence of OROV belonging to Peribunyaviridae. OROV is still localized in the Amazon region and is responsible for causing neurological disease in urban and rural areas [MAYV is endemic in the Amazon region, and there have been imported cases in other regions of Brazil , 44. A pal areas .Flavivirus-positive population and was therefore susceptible to infection. Theoretically, susceptible military personnel could act as disease-spreading agents when returning to civil life when the combination of a susceptible population and specific vectors is present. More detailed serological surveys together with vector population assessment and viral detection strategies are needed to further characterize the extent of favorable factors that can contribute to future outbreaks and to forecast potential public health needs.One of the limitations of the present study is that it was not feasible to perform a neutralization assay to distinguish between dengue serotypes and confirm or rule out multiple infections or cross-reactivity. HIA detects total reactive antibodies against the test antigen; previous studies have demonstrated a lower sensitivity of the HIA compared to ELISA. However, since no sensitivity and specificity values for each virus are available, we could not perform a statistical adjustment to estimate the true prevalence; here, we describe only the crude percentage reactivity for each virus tested in this study. Although most study participants were young male adults, these results are in accordance with previous studies on DENV prevalence , 29. FurAedes aegypti or Culex quinquefasciatus and the decreasing herd immunity to them might facilitate further epidemics of endemic and emerging arboviruses [Aedes and Anopheles mosquito populations [Overall, mosquito control measures and integrated vector management are essential for control of all arboviruses and were effective in controlling ZIKV and CHIKV outbreaks in Brazil and worldwide –50. Howeoviruses , 3. Curroviruses , 14; moroviruses , 51. Warulations .A high percentage of individuals lacked antibodies against major endemic and emerging arboviruses, which makes them susceptible to further infections. Hence, improved vector and febrile syndrome surveillance to identify emerging pathogens is essential to prevent future outbreaks.Additional file 1: Table S1. Socio-demographic features of the study population.Additional file 2: Table S2. Prevalence of antibodies against principal human arboviruses.Additional file 3: Table S3. Dengue seroprevalence in Brazil reported between 1980 and 2020.Additional file 4: Table S4. Distribution of dengue virus serotypes in Brazilian states.Additional file 5: Figure S1. Distribution of dengue, Zika and Chikungunya virus cases in Brazil between 2014 and 2018. Hot spot detection maps were plotted using publicly available data for dengue (a and b), Zika (c and d) and Chikungunya (e and f) virus incidences between 2014 and 2018 . Please note the differences in the incidence rate scales for each virus. North region: Acre: AC, Amapá: AP, Amazonas: AM, Pará: PA, Rondônia: RO, Roraima: RR, Tocantins: TO; Northeast region: Alagoas: AL, Bahia: BA, Ceará: CE, Maranhão: MA, Paraíba: PB, Pernambuco: PE, Piauí: PI, Rio Grande do Norte: RN, Sergipe: SE; Midwest region: Goiás: GO, Mato Grosso: MT, Mato Grosso do Sul: MS, Distrito Federal : DF; Southeast region: Espírito Santo: ES, Minas Gerais: MG, Rio de Janeiro: RJ, São Paulo: SP; South region: Paraná: PR, Rio Grande do Sul: RS, Santa Catarina: SC.


The analyses revealed the infection by a novel betaflexivirus, which tentatively named wheat virus Q (WVQ), together with wheat yellow mosaic virus  and northern cereal mosaic virus (a cytorhabdovirus). Basic local alignment search tool (BLAST) analyses showed that the WVQ strains (of which there are at least three) were related to the members of the genus Foveavirus in the subfamily Quinvirinae (family Betaflexiviridae). In the phylogenetic tree, they form a clade distant from that of the foveaviruses, suggesting that WVQ is a member of a novel genus in the Quinvirinae. Laboratory tests confirmed that WVQ, like WYMV, is potentially transmitted through the soil to wheat plants. WVQ was also found to infect rye plants grown in the same field. Moreover, WVQ-derived small interfering RNAs accumulated in the infected wheat plants, indicating that WVQ infection induces antiviral RNA silencing responses. Given its common coexistence with WYMV, the impact of WVQ infection on yellow mosaic disease in the field warrants detailed investigation.Yellow mosaic disease in winter wheat is usually attributed to the infection by bymoviruses or furoviruses; however, there is still limited information on whether other viral agents are also associated with this disease. To investigate the wheat viromes associated with yellow mosaic disease, we carried out  Triticum aestivum L.) belongs to the grass family Poaceae and is one of the most important crops worldwide. Several diseases caused by vector-borne viruses have significant impacts on cereal crops, including wheat and barley , and furoviruses  are known as disease agents for yellow mosaic disease in wheat and barley and have been distributed widely in wheat/barley-growing regions of many countries  is the causal agent of the wheat yellow mosaic disease and is vectored by the obligate root-inhabiting Benyviridae), wheat yellow striate virus , and European wheat striate mosaic virus  form two phylogenetically separate clades that are assigned to two subfamilies – Trivirinae and Quinvirinae , Banana virus X, and Sugarcane striate mosaic associated virus  for cell-to-cell movement in the leaf epidermis, and are clearly differentiated from the members of the subfamily Trivirinae, whose genomes encode a single movement protein (the 30K superfamily). Most quinviruses belonging to the genera Foveavirus (eight species) and Robigovirus (five species) naturally infect fruit trees, such as apple, cherry, and grapevine, while the members of the genus Carlavirus (53 species) infect various dicotyledonous plants (Allium sativum) plants (via an unknown mechanism (The members of the family nvirinae . The subs plants . Recentl) plants . Some ca) plants . A quinvechanism .de novo meta-RNA-seq approach was used to investigate the virome of wheat plants sampled from a field in Hokkaido, Japan. We discovered three strains of a novel quinvirus tentatively named wheat virus Q (WVQ) that often co-infected wheat with a few known viruses. We also found that WVQ could be transmitted via soil and occurred at least 4 consecutive years in the field, raising the concerns about its potential impact on wheat production.In the current study, a Secale cereale cv. Fuyumidori) samples (leaves and roots) were also collected from the same Naganuma field in 2020 and 2021  in May and June of 2018 and 2019  using default parameters . Subsequently, assembled contigs were used as queries for basic local alignment search tool (BLAST) analyses  against the viral genome reference sequence (RefSeq) dataset of the National Center for Biotechnology Information , and then plasmids were subjected to DNA sequencing.The total RNA was extracted from the plant materials using RNAiso Plus Reagent  according to the manufacturer’s instructions. The cDNA was synthesized using Moloney-murine leukemia virus (MMLV) reverse transcriptase  with random primers , and was then used as a template for PCR amplification with QuickTaq . The conditions of PCR were as follows: 94°C for 2 min; then 30 or 35 cycles of 94°C for 10 s, 53°C or 57°C for 30 s, and 72°C for 1 or 2 min; and 72°C for 10 min. Alternatively, the RNA samples were subjected to one-step reverse transcriptase PCR (RT-PCR) analysis by using PrimeScript One-Step RT-PCR Kit Ver. 2 (TaKaRa), following the manufacturer’s instructions. The 5' and 3' rapid amplification of cDNA ends (5' and 3' RACE) analyses were conducted using the 5'-Full RACE Core Set and 3'-Full RACE Core Set (TaKaRa Bio), respectively, with virus-specific primers . The wheSequence data were analyzed using EnzymeX ver. 3.3.3For the phylogenetic analyses, maximum-likelihood (ML) tree was constructed, as described in Small RNA deep sequencing was performed as described by For soil transmission, the soils that were collected from the field with WYMV-infected wheat plants (KTH-20-1) were used as a virus inoculation source. Commercial culture soil  was also used for non-infested healthy soils. Wheat seeds (cv. “Kitahonami”) were sown in plastic pots (7.5 cm in diameter) containing a mixture of the infested soil and the culture soil, and were grown in a growth cabinet  or in a greenhouse . At different periods after sowing, total RNA extracted from the roots of the plants was subjected to RT-PCR.De novo assembly using the CLC Genomics Workbench yielded 124,777 (pool-18L) and 131,004 (pool-19L) sequence contigs. These assembled contigs were then subjected to local BLAST-N  and BLAST-X (contigs larger than 1.0 kb as queries) analysis against the viral RefSeq collection. At least, 86 contigs  were indicated as candidates for virus-derived sequences. The 41 contigs had a sequence similarity to WYMV , northern cereal mosaic virus , and betaflexiviruses . Approximately, 1.1% of the reads  in pool-18L and 0.9% of the reads  in pool-19 were assigned to virus-related reads in each library. Almost all the virus-associated reads were derived from plant viruses – 43.5%  for WYMV, 0.4%  for NCMV, and 53.3%  for betaflexiviruses, with 2.9%  representing others, likely derived from fungal-associated viruses  were nearly complete sequences of the WYMV RNA1 segment, while the RNA2 coding-complete sequence was generated using partially overlapping contigs in each dataset  and cover nearly the entire region of its genomic RNA (Two negative-sense RNA virus-like contigs (Wh18L_c253 and Wh19L_c20097) were identified from the pool-18L library . These tomic RNA . The conomic RNA  showed aomic RNA  likely romic RNA . The NJ omic RNA . Among tomic RNA . NCMV isomic RNA ; howeverFoveavirus and Carlavirus, subfamily Quinvirinae) using the local BLAST analysis on the two datasets  related to quinviruses . These sequence variants lacked the extra ORF (ORF6) and it seems to be a minor population in the tested wheat sample (KTH-19-1), as the majority of the RACE clones (8/12), which would retain ORF6, had no additional A. The replicase of WVQ does not contain an alkylation B (AlkB)-like domain, which is commonly present in betaflexiviruses that infect perennial plants, such as fruit crops  overlapping the CP gene . The preit crops . They shit crops . Since dit crops , these tit crops  with higvia combinational RT-PCR and direct sequencing. Some overlapping contigs (contigs 936 and 1832 or 935 in pool-19L), likely derived from strain c, show high nucleotide sequence identities >95.9% to each other , indicating the presence of a third strain (designated “strain c”) in the sample pools . A partiQuinvirinae usually have less than about 45% nt identity in their genes  as queries revealed that no significant genome-wide hits were detected . PASC analysis of the genomic sequence of strain a showed 41.1% identity with that of a foveavirus – grapevine rupestris stem pitting-associated virus – which was the top hit sequence . Virusesir genes , suggestir genes . PASCs uir genes ,C. TakenQuinvirinae. WVQ is distantly related to members of the three genera Foveavirus, Carlavirus, and Robigovirus, and other floating or unassigned quinviruses, such as SCSMaV (proposed genus “Sustrivirus”), banana mild mosaic virus (proposed genus “Banmivirus”), garlic yellow mosaic-associated virus, and yam virus Y  were sown in pots containing soils from WYMV-infested fields or commercial soils as a control. One group of pots (three pots for each treatment) being kept under growth-cabinet control at approximately 16°C, which is generally allowed for WYMV infection, while the other group of pots was kept in the greenhouse (non-controlled temperature), where WYMV infection is usually more difficult (Our preliminary finding that WVQ was detected in WYMV-infected roots in the early spring (data not shown) when the temperature was relatively cool and thus the insects usually do not spread widely in the fields, suggests the possibility that WVQ may be transmitted to the roots through soil. Therefore, we aimed to examine the transmission of WVQ ifficult . Under tifficult . After 3ifficult . No systWVQ strains were constantly detected by RT-PCR in the roots and leaves of the wheat plants (cv. “Kitahonami”) that were grown in WYMV-infested fields at Naganuma over 4 consecutive years , tentatively named “wheat virus Q.” Among the three WVQ strains identified in the Naganuma field, two strains were determined on their complete genomic sequences  or formed two or three distinct clades (for CP and TGB1) of foveaviruses , while the underground mechanism, or possible biological vector, of WVQ transmission is still unknown .WVQ is the second quinvirus found to infect cereal crops following SCSMaV, which is distributed across a limited area in Queensland, Australia . WVQ co-graminis  and the  unknown . Interesoviruses . It seemechanism . Only naechanism , while tAspiviridae) and lettuce big-vein associated virus , transmitted by a root-infecting fungus Olpidium brassicae (WVQ was present in at least four consecutive growing seasons in the wheat plants coinfected with WYMV , 6B, andrassicae . MLBVV irassicae . The co-The virus and virus-like sequences derived from this study have been submitted to the GenBank/DDBJ/ENA with the accession numbers LC632066–LC632071.HK designed the experiments. NY and TT collected the samples. HK, NY, MF, KM, KH, HH, and NS performed the experimental work and discussion. HK analyzed the data and wrote the draft manuscript. lBA, NY, TT, and NS were involved in manuscript revision. All authors have given their approval to the 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.