Explore the vast range of titles available.

Human Long interspersed element-1 (L1) retrotransposons contain an internal RNA polymerase II promoter within their 5' untranslated region (UTR) and encode two proteins, (ORF1p and ORF2p) required for their mobilization (i.e., retrotransposition). The evolutionary success of L1 relies on the continuous retrotransposition of full-length L1 mRNAs. Previous studies identified functional splice donor (SD), splice acceptor (SA), and polyadenylation sequences in L1 mRNA and provided evidence that a small number of spliced L1 mRNAs retrotransposed in the human genome. Here, we demonstrate that the retrotransposition of intra-5'UTR or 5'UTR/ORF1 spliced L1 mRNAs leads to the generation of spliced integrated retrotransposed elements (SpIREs). We identified a new intra-5'UTR SpIRE that is ten times more abundant than previously identified SpIREs. Functional analyses demonstrated that both intra-5'UTR and 5'UTR/ORF1 SpIREs lack Cis-acting transcription factor binding sites and exhibit reduced promoter activity. The 5'UTR/ORF1 SpIREs also produce nonfunctional ORF1p variants. Finally, we demonstrate that sequence changes within the L1 5'UTR over evolutionary time, which permitted L1 to evade the repressive effects of a host protein, can lead to the generation of new L1 splicing events, which, upon retrotransposition, generates a new SpIRE subfamily. We conclude that splicing inhibits L1 retrotransposition, SpIREs generally represent evolutionary \"dead-ends\" in the L1 retrotransposition process, mutations within the L1 5'UTR alter L1 splicing dynamics, and that retrotransposition of the resultant spliced transcripts can generate interindividual genomic variation.

LINE-1 (L1) retrotransposons can mobilize (retrotranspose) within the human genome, and mutagenic de novo L1 insertions can lead to human diseases, including cancers. As a result, cells are actively engaged in preventing L1 retrotransposition. This work reveals that the human Condensin II complex restricts L1 retrotransposition in both non-transformed and transformed cell lines through inhibition of L1 transcription and translation. Condensin II subunits, CAP-D3 and CAP-H2, interact with members of the Gamma-Interferon Activated Inhibitor of Translation (GAIT) complex including the glutamyl-prolyl-tRNA synthetase (EPRS), the ribosomal protein L13a, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and NS1 associated protein 1 (NSAP1). GAIT has been shown to inhibit translation of mRNAs encoding inflammatory proteins in myeloid cells by preventing the binding of the translation initiation complex, in response to Interferon gamma (IFN-γ). Excitingly, our data show that Condensin II promotes complexation of GAIT subunits. Furthermore, RNA-Immunoprecipitation experiments in epithelial cells demonstrate that Condensin II and GAIT subunits associate with L1 RNA in a co-dependent manner, independent of IFN-γ. These findings suggest that cooperation between the Condensin II and GAIT complexes may facilitate a novel mechanism of L1 repression, thus contributing to the maintenance of genome stability in somatic cells.

In late 2018, Andes virus was identified as the cause of an outbreak of hantavirus pulmonary syndrome in Argentina. This outbreak was associated with 34 confirmed infections and 11 deaths. A detailed investigation identified person-to-person transmission that was driven by three symptomatic persons.

The real-time generation of information about pathogen genomes has become a vital goal for transmission analysis and characterisation in rapid outbreak responses. In response to the recently established genomic capacity in the Democratic Republic of the Congo, we explored the real-time generation of genomic information at the start of the 2018 Ebola virus disease (EVD) outbreak in North Kivu Province. We used targeted-enrichment sequencing to produce two coding-complete Ebola virus genomes 5 days after declaration of the EVD outbreak in North Kivu. Subsequent sequencing efforts yielded an additional 46 genomes. Genomic information was used to assess early transmission, medical countermeasures, and evolution of Ebola virus. The genomic information demonstrated that the EVD outbreak in the North Kivu and Ituri Provinces was distinct from the 2018 EVD outbreak in Équateur Province of the Democratic Republic of the Congo. Primer and probe mismatches to Ebola virus were identified in silico for all deployed diagnostic PCR assays, with the exception of the Cepheid GeneXpert GP assay. The first two coding-complete genomes provided actionable information in real-time for the deployment of the rVSVΔG-ZEBOV-GP Ebola virus envelope glycoprotein vaccine, available therapeutics, and sequence-based diagnostic assays. Based on the mutations identified in the Ebola virus surface glycoprotein (GP12) observed in all 48 genomes, deployed monoclonal antibody therapeutics (mAb114 and ZMapp) should be efficacious against the circulating Ebola virus variant. Rapid Ebola virus genomic characterisation should be included in routine EVD outbreak response procedures to ascertain efficacy of medical countermeasures. Defense Biological Product Assurance Office.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is causing an exponentially increasing number of coronavirus disease 19 (COVID-19) cases globally. Prioritization of medical countermeasures for evaluation in randomized clinical trials is critically hindered by the lack of COVID-19 animal models that enable accurate, quantifiable, and reproducible measurement of COVID-19 pulmonary disease free from observer bias. We first used serial computed tomography (CT) to demonstrate that bilateral intrabronchial instillation of SARS-CoV-2 into crab-eating macaques ( ) results in mild-to-moderate lung abnormalities qualitatively characteristic of subclinical or mild-to-moderate COVID-19 (e.g., ground-glass opacities with or without reticulation, paving, or alveolar consolidation, peri-bronchial thickening, linear opacities) at typical locations (peripheral>central, posterior and dependent, bilateral, multi-lobar). We then used positron emission tomography (PET) analysis to demonstrate increased FDG uptake in the CT-defined lung abnormalities and regional lymph nodes. PET/CT imaging findings appeared in all macaques as early as 2 days post-exposure, variably progressed, and subsequently resolved by 6-12 days post-exposure. Finally, we applied operator-independent, semi-automatic quantification of the volume and radiodensity of CT abnormalities as a possible primary endpoint for immediate and objective efficacy testing of candidate medical countermeasures.

Long INterspersed Element-1 (LINE-1 or L1) retrotransposons are an ancient family of repeated DNA sequences present in all inspected mammalian genomes. L1s are the only active autonomous mobile element in the human genome and are present at over 500,000 copies, representing ~17% of genomic DNA. A full-length human L1 is ~6 kb in length and contains an internal RNA polymerase II promoter within its 5’ untranslated region (UTR). Following the 5’UTR are two open reading frames (ORF1 and ORF2) that encode two functional proteins (ORF1p and ORF2p) that are required for mobilization (i.e., retrotransposition). L1s end with a 3’UTR and a poly(A) tail. The evolutionary success of L1 relies on the reiterative retrotransposition of full-length L1 RNAs. The vast majority (>99.9%) of genomic L1s are inactive; however, on average, ~80-100 L1s per diploid genome are capable of retrotransposition. Since, L1 retrotransposition by its nature is mutagenic, it is likely that cellular host-factors have evolved to inhibit or restrict unabated L1 retrotransposition. Previous studies identified functional splice donor, splice acceptor, and polyadenylation sequences in full-length L1 RNA. Here, I demonstrate that retrotransposition of intra-5’UTR or 5’UTR/ORF1 spliced L1 RNAs leads to the generation of Spliced Integrated Retrotransposed Elements (SpIREs). Additionally, I uncovered a new intra-5’UTR SpIRE that is approximately ten times more abundant than previously identified SpIREs. Using biochemical and genetic approaches, I definitively demonstrate that intra-5’UTR SpIREs lack cis-acting transcription factor binding sites, resulting in reduced 5’UTR promoter activity compared to a full-length 5’UTR. Moreover, I demonstrate that 5’UTR/ORF1 SpIREs lack cis-acting sequences required for L1 transcription and produce non-functional ORF1p variants. These results establish that SpIREs are evolutionary “dead ends,” which are unlikely to contribute to additional rounds of L1 retrotransposition. Finally, in agreement with previous publications, I demonstrate that a subset of splicing factors may repress L1 expression and/or retrotransposition. Previous experiments in embryonic human carcinoma-derived cells (hECs) revealed that an engineered L1 tagged with a retrotransposition indicator cassette (L1- reporter) successfully retrotransposed into hEC genomic DNA. However, either during or immediately after genomic integration, expression of the L1-reporter is silenced in hECs. Previous experiments demonstrated that hEC cells treated with histone deacetylase (HDAC) inhibitors swiftly reverse L1-reporter silencing. I sought to identify host proteins that may be involved in L1-reporter silencing by developing a forward genetic screen using CRISPR/Cas9-based genome editing technology. Using the PA-1 hEC cell line that permits L1-reporter retrotransposition, but subsequently silences L1- reporter expression, I identified potential candidate genes that may play a role in L1- reporter mediated gene silencing. Future work will test whether the candidate genes identified in this screen directly or indirectly inhibit L1-reporter mediated gene silencing. A continued understanding of the interplay between L1 and host-factors is critical to understanding human genomic variation. Additionally, as the L1-host interaction largely recapitulates a traditional host-parasite “arms-race,” new insights gained from these studies may contribute to our understanding of how L1 retrotransposition continues to influence human health and disease.

Introduction Aerosol pollutants are known to raise the risk of development of non-communicable respiratory diseases (NCRDs) such as asthma, chronic bronchitis, chronic obstructive pulmonary disease, and allergic rhinitis. Sub-Saharan Africa’s rapid pace of urbanization, economic expansion, and population growth raise concerns of increasing incidence of NCRDs. This research characterizes the state of research on pollution and NCRDs in the 46 countries of Sub-Saharan Africa (SSA). This research systematically reviewed the literature on studies of asthma; chronic bronchitis; allergic rhinitis; and air pollutants such as particulate matter, ozone, NOx, and sulfuric oxide. Methods We searched three major databases (PubMed, Web of Science, and Scopus) using the key words “asthma”, “chronic bronchitis”, “allergic rhinitis”, and “COPD” with “carbon monoxide (CO)”, “sulfuric oxide (SO)”, “ozone (O3)”, “nitrogen dioxide (NO2)”, and “particulate matter (PM)”, restricting the search to the 46 countries that comprise SSA. Only papers published in scholarly journals with a defined health outcome in individuals and which tested associations with explicitly measured or modelled air exposures were considered for inclusion. All candidate papers were entered into a database for review. Results We found a total of 362 unique research papers in the initial search of the three databases. Among these, 14 met the inclusion criteria. These papers comprised studies from just five countries. Nine papers were from South Africa; two from Malawi; and one each from Ghana, Namibia, and Nigeria. Most studies were cross-sectional. Exposures to ambient air pollutants were measured using spectrometry and chromatography. Some studies created composite measures of air pollution using a range of data layers. NCRD outcomes were measured by self-reported health status and measures of lung function (spirometry). Populations of interest were primarily schoolchildren, though a few studies focused on secondary school students and adults. Conclusions The paucity of research on NCRDs and ambient air pollutant exposures is pronounced within the African continent. While capacity to measure air quality in SSA is high, studies targeting NCRDs should work to draw attention to questions of outdoor air pollution and health. As the climate changes and SSA economies expand and countries urbanize, these questions will become increasingly important.

Introduction Snakebites are a major cause of permanent injury and death among poor, rural populations in developing countries, including those in East Africa. This research characterizes snakebite incidence, risk factors, and subsequent health-seeking behaviors in two regions of Kenya using a mixed methods approach. Methods As a part of regular activities of a health demographic surveillance system, household-level survey on snakebite incidence was conducted in two areas of Kenya: Kwale along the Kenyan Coast and Mbita on Lake Victoria. If someone in the home was reported to have been bitten in the 5 years previous to the visit, a survey instrument was administered. The survey gathered contextual information on the bite, treatment-seeking behavior and clinical manifestations. To obtain deeper, contextual information, respondents were also asked to narrate the bite incident, subsequent behavior and outcomes. Results 8775 and 9206 households were surveyed in Kwale and Mbita, respectively. Out of these, 453 (5.17%) and 92 (1.00%) households reported that at least one person had been bitten by a snake in the past 5 years. Deaths from snakebites were rare (4.04%), but patterns of treatment seeking varied. Treatment at formal care facilities were sought for 50.8% and at traditional healers for 53.3%. 18.4% sought treatment from both sources. Victims who delayed receiving treatment from a formal facility were more likely to have consulted a traditional healer (OR 8.8995% CI [3.83, 20.64]). Delays in treatment seeking were associated with significantly increased odds of having a severe outcome, including death, paralysis or loss of consciousness (OR 3.47 95% CI [1.56; 7.70]). Conclusion Snakebite incidence and outcomes vary by region in Kenya, and treatment-seeking behaviors are complex. Work needs to be done to better characterize the spatial distribution of snakebite incidence in Kenya and efforts need to be made to ensure that victims have sufficient access to effective treatments to prevent death and serious injury.