ChimeraTE: a pipeline to detect chimeric transcripts derived from genes and transposable elements.

Transposable elements (TEs) produce structural variants and are considered an important source of genetic diversity. Notably, TE-gene fusion transcripts, i.e. chimeric transcripts, have been associated with adaptation in several species. However, the identification of these chimeras remains hindered due to the lack of detection tools at a transcriptome-wide scale, and to the reliance on a reference genome, even though different individuals/cells/strains have different TE insertions. Therefore, we developed ChimeraTE, a pipeline that uses paired-end RNA-seq reads to identify chimeric transcripts through two different modes. Mode 1 is the reference-guided approach that employs canonical genome alignment, and Mode 2 identifies chimeras derived from fixed or insertionally polymorphic TEs without any reference genome. We have validated both modes using RNA-seq data from four Drosophila melanogaster wild-type strains. We found ∼1.12% of all genes generating chimeric transcripts, most of them from TE-exonized sequences. Approximately ∼23% of all detected chimeras were absent from the reference genome, indicating that TEs belonging to chimeric transcripts may be recent, polymorphic insertions. ChimeraTE is the first pipeline able to automatically uncover chimeric transcripts without a reference genome, consisting of two running Modes that can be used as a tool to investigate the contribution of TEs to transcriptome plasticity.

SEDA 2024 update: enhancing the SEquence DAtaset builder for seamless integration into automated data analysis pipelines.

BACKGROUND: The initial version of SEDA assists life science researchers without programming skills with the preparation of DNA and protein sequence FASTA files for multiple bioinformatics applications. However, the initial version of SEDA lacks a command-line interface for more advanced users and does not allow the creation of automated analysis pipelines. RESULTS: The present paper discusses the updates of the new SEDA release, including the addition of a complete command-line interface, new functionalities like gene annotation, a framework for automated pipelines, and improved integration in Linux environments. CONCLUSION: SEDA is an open-source Java application and can be installed using the different distributions available ( https://www.sing-group.org/seda/download.html ) as well as through a Docker image ( https://hub.docker.com/r/pegi3s/seda ). It is released under a GPL-3.0 license, and its source code is publicly accessible on GitHub ( https://github.com/sing-group/seda ). The software version at the time of submission is archived at Zenodo (version v1.6.0, http://doi.org/10.5281/zenodo.10201605 ).

Multiple Lines of Evidence Support 199 SARS-CoV-2 Positively Selected Amino Acid Sites.

SARS-CoV-2 amino acid variants that contribute to an increased transmissibility or to host immune system escape are likely to increase in frequency due to positive selection and may be identified using different methods, such as codeML, FEL, FUBAR, and MEME. Nevertheless, when using different methods, the results do not always agree. The sampling scheme used in different studies may partially explain the differences that are found, but there is also the possibility that some of the identified positively selected amino acid sites are false positives. This is especially important in the context of very large-scale projects where hundreds of analyses have been performed for the same protein-coding gene. To account for these issues, in this work, we have identified positively selected amino acid sites in SARS-CoV-2 and 15 other coronavirus species, using both codeML and FUBAR, and compared the location of such sites in the different species. Moreover, we also compared our results to those that are available in the COV2Var database and the frequency of the 10 most frequent variants and predicted protein location to identify those sites that are supported by multiple lines of evidence. Amino acid changes observed at these sites should always be of concern. The information reported for SARS-CoV-2 can also be used to identify variants of concern in other coronaviruses.

Maternal and perinatal risk factors associated with congenital syphilis.

OBJECTIVE: Mother-to-child transmission of syphilis remains an important global public health problem. Untreated intrauterine infection may result in adverse events for the fetus or newborn (NB). Maternal risk factors, such as prenatal care, early diagnosis, and appropriate treatment, significantly impact the likelihood of vertical transmission of syphilis. The purpose of this review is to evaluate maternal risk factors for congenital syphilis and the characteristics of exposed NB. METHODS: A total of 14 studies were evaluated, including 8 cohort studies, 4 cross-sectional and 2 control cases. A total of 12,230 women were included, with confirmed or highly probable congenital syphilis outcome, and 2285 NB. The studies evaluated risk factors for congenital syphilis, which were maternal, demographic, obstetric factors and characteristics of the exposed NB. RESULTS: Included in the risk factors studied, inadequate prenatal care and late onset, as well as inadequate or late treatment of maternal syphilis were significant risk factors for the outcome of congenital syphilis. When the time set of maternal diagnosis was correlated with neonatal infection, there was a tendency to worsen prognosis (more infected NB) in women diagnosed later in pregnancy, as well as in women who underwent few prenatal consultations and inadequate treatment. Women with recent syphilis with high VDRL titres had a higher rate of vertical transmission. The prior history of syphilis with adequate treatment was identified as a protective factor, resulting in lower rates of congenital syphilis. Among the epidemiological and demographic aspects surveyed, it was observed that young age, lower schooling, unemployment, low family income and no fixed residence were associated with higher risk of congenital syphilis. CONCLUSIONS: The association of syphilis with adverse socio-economic conditions and inadequate prenatal care suggests that the improvement of the population's living conditions and equitable access to quality health services may have an impact on the reduction of congenital syphilis.

Labour market participation among rehabilitated individuals with spinal cord injury in Brazil: a cross-sectional study.

STUDY DESIGN: Cross-sectional study. OBJECTIVE: To document the labour market participation rate and investigate the impact of social and economic characteristics on this outcome among individuals with spinal cord injury or disease (SCI/D) who participated in a multidisciplinary rehabilitation programme in Brazil. SETTING: A specialised rehabilitation centre in southern Brazil. METHODS: An interview was performed with former rehabilitation clients with SCI/D. A total of 111 community-dwelling individuals with SCI/D who had been previously rehabilitated in a large regional rehabilitation centre were considered. Employment status and demographic, injury, work, and economic characteristics were self-reported via interview. Descriptive statistical analyses were performed. RESULTS: The labour market participation rate was 21.6% (24 individuals). Five respondents were employed in formal work activities, and the other 19 were involved in informal activities (self-employed). Since the onset of injury, a shorter time was associated with nonparticipation in occupational work. Individuals with a higher level of education were more likely to be working. Household income was higher among the participants who had returned to work than among those who had not returned. CONCLUSION: There is a relatively low labour market participation rate among rehabilitated individuals with SCI/D in southern Brazil. The rehabilitation services should emphasise vocational training, access to education, and employment support for individuals with SCI/D from early onset after the injury. Public policies must be revised to support labour market participation among individuals with SCI/D in Brazil.

The Worldwide Invasion of Drosophila suzukii Is Accompanied by a Large Increase of Transposable Element Load and a Small Number of Putatively Adaptive Insertions.

Transposable elements (TEs) are ubiquitous and mobile repeated sequences. They are major determinants of host fitness. Here, we characterized the TE content of the spotted wing fly Drosophila suzukii. Using a recently improved genome assembly, we reconstructed TE sequences de novo and found that TEs occupy 47% of the genome and are mostly located in gene-poor regions. The majority of TE insertions segregate at low frequencies, indicating a recent and probably ongoing TE activity. To explore TE dynamics in the context of biological invasions, we studied the variation of TE abundance in genomic data from 16 invasive and six native populations of D. suzukii. We found a large increase of the TE load in invasive populations correlated with a reduced Watterson estimate of genetic diversity θw^ a proxy of effective population size. We did not find any correlation between TE contents and bioclimatic variables, indicating a minor effect of environmentally induced TE activity. A genome-wide association study revealed that ca. 2,000 genomic regions are associated with TE abundance. We did not find, however, any evidence in such regions of an enrichment for genes known to interact with TE activity (e.g., transcription factor encoding genes or genes of the piRNA pathway). Finally, the study of TE insertion frequencies revealed 15 putatively adaptive TE insertions, six of them being likely associated with the recent invasion history of the species.

Evaluation of the lights4violence program: reduction in machismo and acceptance of violence among adolescents in Europe.

BACKGROUND: Machismo and acceptance of violence (AV) against women are part of the social construction of hegemonic masculinity and are related to the risk of dating violence. This study aims to analyze the effectiveness of the Lights4Violence program in reducing machismo and AV in secondary school students from different European cities. METHODS: Quasi-experimental longitudinal study using a convenience sample of 1,146 high school students from different European cities (12-17 years old) including 575 intervention group students (59.1% girls) and 571 control group students (62.7% girls). We performed linear regression models to identify the effect of the intervention, modelling the difference in means in machismo and AV (dependent variables) between wave-2 and wave-1. RESULTS: An interaction was identified between the group variable and the empathy variable. In wave-2, girls with high empathy at baseline in the intervention group obtained lower mean AV scores (ß: -0.131; p = 0.004). However, the boys in the intervention group (reference: control group) with low empathy at baseline registered a significant increase in the mean values of machismo (ß: 0.247; p < 0.001). CONCLUSION: The importance of empathy is shown in the effectiveness of interventions to reduce machismo and AV in adolescents. While the Lights4Violence program focuses on promoting healthy relationships, there were some controversial results. It is possible that some children, especially those with less empathy, may have felt "challenged" during the intervention and/or assessment. This suggests the need for the development of interventions that also consider psychological processes and integrate the promotion of positive expressions of masculinity.

The transposable element-rich genome of the cereal pest Sitophilus oryzae.

BACKGROUND: The rice weevil Sitophilus oryzae is one of the most important agricultural pests, causing extensive damage to cereal in fields and to stored grains. S. oryzae has an intracellular symbiotic relationship (endosymbiosis) with the Gram-negative bacterium Sodalis pierantonius and is a valuable model to decipher host-symbiont molecular interactions. RESULTS: We sequenced the Sitophilus oryzae genome using a combination of short and long reads to produce the best assembly for a Curculionidae species to date. We show that S. oryzae has undergone successive bursts of transposable element (TE) amplification, representing 72% of the genome. In addition, we show that many TE families are transcriptionally active, and changes in their expression are associated with insect endosymbiotic state. S. oryzae has undergone a high gene expansion rate, when compared to other beetles. Reconstruction of host-symbiont metabolic networks revealed that, despite its recent association with cereal weevils (30 kyear), S. pierantonius relies on the host for several amino acids and nucleotides to survive and to produce vitamins and essential amino acids required for insect development and cuticle biosynthesis. CONCLUSIONS: Here we present the genome of an agricultural pest beetle, which may act as a foundation for pest control. In addition, S. oryzae may be a useful model for endosymbiosis, and studying TE evolution and regulation, along with the impact of TEs on eukaryotic genomes.

Evidence of cross-contamination of waste workers and transmission of antimicrobial resistance genes by coagulase-negative Staphylococcus isolated from dental solid waste: an intriguing study.

The aim of this study was to phenotypically and genotypically identify coagulase-negative Staphylococcus (CoNS) recovered from the nostrils of waste workers and from dental waste; 135 strains were recovered and S. epidermidis was the prevailing species. Genetic similarity (100%) was observed between the two S. epidermidis isolated from different employees on the same shift and 85% similarity between the S. epidermidis recovered from an employee's nostril and from waste. The mecA gene was found in 20 CoNS, and 20% were also found to possess the vanA gene. The blaZ gene was detected in 46.7%, and the icaA (34.8%), B and C genes (11.8% each). Our findings emphasized the biological risk to which waste workers are exposed and unprecedently confirms that it was possible to recover genetically identical bacterial species from waste and from workers' nostrils. It is important to highlight that this risk is raised by the detection of relevant antimicrobial resistance genes. The results also suggest that effective measures to correctly manage waste and promote the rational use of antimicrobials should be adopted.

Genomic Analysis of European Drosophila melanogaster Populations Reveals Longitudinal Structure, Continent-Wide Selection, and Previously Unknown DNA Viruses.

Genetic variation is the fuel of evolution, with standing genetic variation especially important for short-term evolution and local adaptation. To date, studies of spatiotemporal patterns of genetic variation in natural populations have been challenging, as comprehensive sampling is logistically difficult, and sequencing of entire populations costly. Here, we address these issues using a collaborative approach, sequencing 48 pooled population samples from 32 locations, and perform the first continent-wide genomic analysis of genetic variation in European Drosophila melanogaster. Our analyses uncover longitudinal population structure, provide evidence for continent-wide selective sweeps, identify candidate genes for local climate adaptation, and document clines in chromosomal inversion and transposable element frequencies. We also characterize variation among populations in the composition of the fly microbiome, and identify five new DNA viruses in our samples.

Oxidative and radiation stress induces transposable element transcription in Drosophila melanogaster.

It has been shown that stressors are capable of activating transposable elements (TEs). Currently, there is a hypothesis that stress activation of TEs may be involved in adaptive evolution, favouring the increase in genetic variability when the population is under adverse conditions. However, TE activation under stress is still poorly understood. In the present study, we estimated the fraction of differentially expressed TEs (DETEs) under ionizing radiation (144, 360 and 864 Gy) and oxidative stress (dioxin, formaldehyde and toluene) treatments. The stress intensity of each treatment was estimated by measuring the number of differentially expressed genes, and we show that several TEs families are activated by stress whereas others are repressed. The proportion of DETEs was positively related to stress intensity. However, even under the strongest stress, only a small fraction of TE families were activated (9.28%) and 17.72% were repressed. Considering all treatments together, the activated proportion was 19.83%. Nevertheless, as several TEs are incomplete or degenerated, only 10.55% of D. melanogaster mobilome is, at same time, activated by the stressors and able to transpose or at least code a protein. Thus, our study points out that although stress activates TEs, it is not a generalized activation process, and for some families, the stress induces repression.

Less effective selection leads to larger genomes.

The evolutionary origin of the striking genome size variations found in eukaryotes remains enigmatic. The effective size of populations, by controlling selection efficacy, is expected to be a key parameter underlying genome size evolution. However, this hypothesis has proved difficult to investigate using empirical data sets. Here, we tested this hypothesis using 22 de novo transcriptomes and low-coverage genomes of asellid isopods, which represent 11 independent habitat shifts from surface water to resource-poor groundwater. We show that these habitat shifts are associated with higher transcriptome-wide [Formula: see text] After ruling out the role of positive selection and pseudogenization, we show that these transcriptome-wide [Formula: see text] increases are the consequence of a reduction in selection efficacy imposed by the smaller effective population size of subterranean species. This reduction is paralleled by an important increase in genome size (25% increase on average), an increase also confirmed in subterranean decapods and mollusks. We also control for an adaptive impact of genome size on life history traits but find no correlation between body size, or growth rate, and genome size. We show instead that the independent increases in genome size measured in subterranean isopods are the direct consequence of increasing invasion rates by repeat elements, which are less efficiently purged out by purifying selection. Contrary to selection efficacy, polymorphism is not correlated to genome size. We propose that recent demographic fluctuations and the difficulty of observing polymorphism variation in polymorphism-poor species can obfuscate the link between effective population size and genome size when polymorphism data are used alone.

Multiple independent L-gulonolactone oxidase (GULO) gene losses and vitamin C synthesis reacquisition events in non-Deuterostomian animal species.

BACKGROUND: L-ascorbate (Vitamin C) is an important antioxidant and co-factor in eukaryotic cells, and in mammals it is indispensable for brain development and cognitive function. Vertebrates usually become L-ascorbate auxothrophs when the last enzyme of the synthetic pathway, an L-gulonolactone oxidase (GULO), is lost. Since Protostomes were until recently thought not to have a GULO gene, they were considered to be auxothrophs for Vitamin C. RESULTS: By performing phylogenetic analyses with tens of non-Bilateria and Protostomian genomes, it is shown, that a GULO gene is present in the non-Bilateria Placozoa, Myxozoa (here reported for the first time) and Anthozoa groups, and in Protostomians, in the Araneae family, the Gastropoda class, the Acari subclass (here reported for the first time), and the Priapulida, Annelida (here reported for the first time) and Brachiopoda phyla lineages. GULO is an old gene that predates the separation of Animals and Fungi, although it could be much older. We also show that within Protostomes, GULO has been lost multiple times in large taxonomic groups, namely the Pancrustacea, Nematoda, Platyhelminthes and Bivalvia groups, a pattern similar to that reported for Vertebrate species. Nevertheless, we show that Drosophila melanogaster seems to be capable of synthesizing L-ascorbate, likely through an alternative pathway, as recently reported for Caenorhabditis elegans. CONCLUSIONS: Non-Bilaterian and Protostomians seem to be able to synthesize Vitamin C either through the conventional animal pathway or an alternative pathway, but in this animal group, not being able to synthesize L-ascorbate seems to be the exception rather than the rule.

Population-specific dynamics and selection patterns of transposable element insertions in European natural populations.

Transposable elements (TEs) are ubiquitous sequences in genomes of virtually all species. While TEs have been investigated for several decades, only recently we have the opportunity to study their genome-wide population dynamics. Most of the studies so far have been restricted either to the analysis of the insertions annotated in the reference genome or to the analysis of a limited number of populations. Taking advantage of the European Drosophila population genomics consortium (DrosEU) sequencing data set, we have identified and measured the dynamics of TEs in a large sample of European Drosophila melanogaster natural populations. We showed that the mobilome landscape is population-specific and highly diverse depending on the TE family. In contrast with previous studies based on SNP variants, no geographical structure was observed for TE abundance or TE divergence in European populations. We further identified de novo individual insertions using two available programs and, as expected, most of the insertions were present at low frequencies. Nevertheless, we identified a subset of TEs present at high frequencies and located in genomic regions with a high recombination rate. These TEs are candidates for being the target of positive selection, although neutral processes should be discarded before reaching any conclusion on the type of selection acting on them. Finally, parallel patterns of association between the frequency of TE insertions and several geographical and temporal variables were found between European and North American populations, suggesting that TEs can be potentially implicated in the adaptation of populations across continents.

TEtools facilitates big data expression analysis of transposable elements and reveals an antagonism between their activity and that of piRNA genes.

Over recent decades, substantial efforts have been made to understand the interactions between host genomes and transposable elements (TEs). The impact of TEs on the regulation of host genes is well known, with TEs acting as platforms of regulatory sequences. Nevertheless, due to their repetitive nature it is considerably hard to integrate TE analysis into genome-wide studies. Here, we developed a specific tool for the analysis of TE expression: TEtools. This tool takes into account the TE sequence diversity of the genome, it can be applied to unannotated or unassembled genomes and is freely available under the GPL3 (https://github.com/l-modolo/TEtools). TEtools performs the mapping of RNA-seq data obtained from classical mRNAs or small RNAs onto a list of TE sequences and performs differential expression analyses with statistical relevance. Using this tool, we analyzed TE expression from five Drosophila wild-type strains. Our data show for the first time that the activity of TEs is strictly linked to the activity of the genes implicated in the piwi-interacting RNA biogenesis and therefore fits an arms race scenario between TE sequences and host control genes.

An attempt to select non-genetic variation in resistance to starvation and reduced chill coma recovery time in Drosophila melanogaster.

Phenotypic variance is attributed to genetic and non-genetic factors, and only the former are presumed to be inherited and thus suitable for the action of selection. Although increasing amounts of data suggest that non-genetic variability may be inherited, we have limited empirical data in animals. Here, we performed an artificial selection experiment using Drosophila melanogaster inbred lines. We quantified the response to selection for a decrease in chill coma recovery time and an increase in starvation resistance. We observed a weak response to selection in the inbred and outbred lines, with variability across lines. At the end of the selection process, differential expression was detected for some genes associated with epigenetics, the piRNA pathway and canalization functions. As the selection process can disturb the canalization process and increase the phenotypic variance of developmental traits, we also investigated possible effects of the selection process on the number of scutellar bristles, fluctuating asymmetry levels and fitness estimates. These results suggest that, contrary to what was shown in plants, selection of non-genetic variability is not straightforward in Drosophila and appears to be strongly genotype dependent.

SNP calling from RNA-seq data without a reference genome: identification, quantification, differential analysis and impact on the protein sequence.

SNPs (Single Nucleotide Polymorphisms) are genetic markers whose precise identification is a prerequisite for association studies. Methods to identify them are currently well developed for model species, but rely on the availability of a (good) reference genome, and therefore cannot be applied to non-model species. They are also mostly tailored for whole genome (re-)sequencing experiments, whereas in many cases, transcriptome sequencing can be used as a cheaper alternative which already enables to identify SNPs located in transcribed regions. In this paper, we propose a method that identifies, quantifies and annotates SNPs without any reference genome, using RNA-seq data only. Individuals can be pooled prior to sequencing, if not enough material is available from one individual. Using pooled human RNA-seq data, we clarify the precision and recall of our method and discuss them with respect to other methods which use a reference genome or an assembled transcriptome. We then validate experimentally the predictions of our method using RNA-seq data from two non-model species. The method can be used for any species to annotate SNPs and predict their impact on the protein sequence. We further enable to test for the association of the identified SNPs with a phenotype of interest.

High-throughput sequencing of transposable element insertions suggests adaptive evolution of the invasive Asian tiger mosquito towards temperate environments.

Invasive species represent unique opportunities to evaluate the role of local adaptation during colonization of new environments. Among these species, the Asian tiger mosquito, Aedes albopictus, is a threatening vector of several human viral diseases, including dengue and chikungunya, and raises concerns about the Zika fever. Its broad presence in both temperate and tropical environments has been considered the reflection of great "ecological plasticity." However, no study has been conducted to assess the role of adaptive evolution in the ecological success of Ae. albopictus at the molecular level. In the present study, we performed a genomic scan to search for potential signatures of selection leading to local adaptation in one-hundred-forty field-collected mosquitoes from native populations of Vietnam and temperate invasive populations of Europe. High-throughput genotyping of transposable element insertions led to the discovery of more than 120,000 polymorphic loci, which, in their great majority, revealed a virtual absence of structure between the biogeographic areas. Nevertheless, 92 outlier loci showed a high level of differentiation between temperate and tropical populations. The majority of these loci segregate at high insertion frequencies among European populations, indicating that this pattern could have been caused by recent adaptive evolution events in temperate areas. An analysis of the overlapping and neighbouring genes highlighted several candidates, including diapause, lipid and juvenile hormone pathways.

Description of new genera and species of marine cyanobacteria from the Portuguese Atlantic coast.

Aiming at increasing the knowledge on marine cyanobacteria from temperate regions, we previously isolated and characterized 60 strains from the Portuguese foreshore and evaluate their potential to produce secondary metabolites. About 15% of the obtained 16S rRNA gene sequences showed less than 97% similarity to sequences in the databases revealing novel biodiversity. Herein, seven of these strains were extensively characterized and their classification was re-evaluated. The present study led to the proposal of five new taxa, three genera (Geminobacterium, Lusitaniella, and Calenema) and two species (Hyella patelloides and Jaaginema litorale). Geminobacterium atlanticum LEGE 07459 is a chroococcalean that shares morphological characteristics with other unicellular cyanobacterial genera but has a distinct phylogenetic position and particular ultrastructural features. The description of the Pleurocapsales Hyella patelloides LEGE 07179 includes novel molecular data for members of this genus. The filamentous isolates of Lusitaniella coriacea - LEGE 07167, 07157 and 06111 - constitute a very distinct lineage, and seem to be ubiquitous on the Portuguese coast. Jaaginema litorale LEGE 07176 has distinct characteristics compared to their marine counterparts, and our analysis indicates that this genus is polyphyletic. The Synechococcales Calenema singularis possess wider trichomes than Leptolyngbya, and its phylogenetic position reinforces the establishment of this new genus.

Knowledge, behaviour and microbial load of workers handling dental solid waste in a public health service in Brazil.

Healthcare wastes are those generated inside healthcare services, including dental clinics. Workers coming into close proximity to hazardous healthcare waste are potentially at risk. In an attempt to assess the knowledge and attitudes of workers dealing with infectious waste, a questionnaire was administered. The biological risk was investigated by evaluating the microbial load and screening some clinically relevant micro-organisms in the nasal mucosa, hands and coats of these workers. The results showed that 66.6% of the study population had incomplete primary education. Only two workers have had their blood tested to confirm anti-HBs levels. Microbial load evaluation on hand surfaces of morning workers showed statistically significant lower microbial loads after the workday when compared with the beginning of the work period. It is important to highlight that some clinically relevant bacteria, such as Pseudomonas aeruginosa and Klebsiella pneumoniae were isolated from worker's hands. This study revealed the need for more training programmes regarding awareness of safe waste disposal protocols and also the necessity of discussing vaccination and its implications. Data regarding microbial loads of the worker's hands, mostly at the beginning of the workday when handwashing is recommended worldwide, emphasise that hygiene measures should receive more attention during training exercises.