The DEAD box RNA helicase DDX42 is an intrinsic inhibitor of positive-strand RNA viruses.

Genome-wide screens are powerful approaches to unravel regulators of viral infections. Here, a CRISPR screen identifies the RNA helicase DDX42 as an intrinsic antiviral inhibitor of HIV-1. Depletion of endogenous DDX42 increases HIV-1 DNA accumulation and infection in cell lines and primary cells. DDX42 overexpression inhibits HIV-1 infection, whereas expression of a dominant-negative mutant increases infection. Importantly, DDX42 also restricts LINE-1 retrotransposition and infection with other retroviruses and positive-strand RNA viruses, including CHIKV and SARS-CoV-2. However, DDX42 does not impact the replication of several negative-strand RNA viruses, arguing against an unspecific effect on target cells, which is confirmed by RNA-seq analysis. Proximity ligation assays show DDX42 in the vicinity of viral elements, and cross-linking RNA immunoprecipitation confirms a specific interaction of DDX42 with RNAs from sensitive viruses. Moreover, recombinant DDX42 inhibits HIV-1 reverse transcription in vitro. Together, our data strongly suggest a direct mode of action of DDX42 on viral ribonucleoprotein complexes. Our results identify DDX42 as an intrinsic viral inhibitor, opening new perspectives to target the life cycle of numerous RNA viruses.

Epitranscriptomic regulation of HIV-1 full-length RNA packaging.

During retroviral replication, the full-length RNA serves both as mRNA and genomic RNA. However, the mechanisms by which the HIV-1 Gag protein selects the two RNA molecules that will be packaged into nascent virions remain poorly understood. Here, we demonstrate that deposition of N6-methyladenosine (m6A) regulates full-length RNA packaging. While m6A deposition by METTL3/METTL14 onto the full-length RNA was associated with increased Gag synthesis and reduced packaging, FTO-mediated demethylation promoted the incorporation of the full-length RNA into viral particles. Interestingly, HIV-1 Gag associates with the RNA demethylase FTO in the nucleus and contributes to full-length RNA demethylation. We further identified two highly conserved adenosines within the 5'-UTR that have a crucial functional role in m6A methylation and packaging of the full-length RNA. Together, our data propose a novel epitranscriptomic mechanism allowing the selection of the HIV-1 full-length RNA molecules that will be used as viral genomes.

The effect of height on family formation in rural Spain, birth-cohorts 1835-1975.

This article examines the relationship between the height of adult males and marital outcomes, including likelihood of marrying, age at marriage, and marital fertility, in rural Spain. For this analysis, a sample of 4,501 men born between 1835 and 1975 living in 14 villages in northeastern Spain was taken. Previous research has shown that shorter individuals are less likely to marry. However, it is still disputed whether differences exist in the timing of marrying based on height, and little attention has been paid to the effect(s) of height on offspring. Family data were obtained from parish records and interviews with individuals and their families, while height data were obtained from military records, with individuals in Spain being conscripted at the age of 21 years. The data were linked according to nominative criteria using family reconstitution methods. The results confirm that shorter individuals were less likely to marry. Individuals of medium and medium-high height were the first to marry, with a small gap between them and shorter individuals. With regard to marital fertility, no difference in terms of average fertility by height were found, but there were small differences in timing of childbirth, possibly as a result of delayed marriage.

CBP80/20-dependent translation initiation factor (CTIF) inhibits HIV-1 Gag synthesis by targeting the function of the viral protein Rev.

Translation initiation of the human immunodeficiency virus type-1 (HIV-1) full-length RNA has been shown to occur through cap-dependent and IRES-driven mechanisms. Previous studies suggested that the nuclear cap-binding complex (CBC) rather than eIF4E drives cap-dependent translation of the full-length RNA and we have recently reported that the CBC subunit CBP80 supports the function of the viral protein Rev during nuclear export and translation of this viral transcript. Ribosome recruitment during CBC-dependent translation of cellular mRNAs relies on the activity CBP80/20 translation initiation factor (CTIF), which bridges CBP80 and the 40S ribosomal subunit through interactions with eIF3g. Here, we report that CTIF inhibits HIV-1 and HIV-2 Gag synthesis from the full-length RNA. Our results indicate that CTIF associates with HIV-1 Rev through its N-terminal domain and is recruited onto the full-length RNA ribonucleoprotein complex in order to interfere with Gag synthesis. We also demonstrate that CTIF induces the cytoplasmic accumulation of Rev impeding the association of the viral protein with CBP80. We finally show that Rev interferes with the association of CTIF with CBP80 indicating that CTIF and Rev compete for the CBC subunit.

Sustained hyperammonemia induces TNF-a IN Purkinje neurons by activating the TNFR1-NF-κB pathway.

BACKGROUND: Patients with liver cirrhosis may develop hepatic encephalopathy. Rats with chronic hyperammonemia exhibit neurological alterations mediated by peripheral inflammation and neuroinflammation. Motor incoordination is due to increased TNF-a levels and activation of its receptor TNFR1 in the cerebellum. The aims were to assess (a) whether peripheral inflammation is responsible for TNF-a induction in hyperammonemic rats, (b) the cell type(s) in which TNF-a is increased, (c) whether this increase is associated with increased nuclear NF-κB and TNFR1 activation, (d) the time course of TNF-a induction, and (e) if TNF-a is induced in the Purkinje neurons of patients who die with liver cirrhosis. METHODS: We analyzed the level of TNF-a mRNA and NF-κB in microglia, astrocytes, and Purkinje neurons in the cerebellum after 1, 2, and 4 weeks of hyperammonemia. We assessed whether preventing peripheral inflammation by administering an anti-TNF-a antibody prevents TNF-a induction. We tested whether TNF-a induction is reversed by R7050, which inhibits the TNFR1-NF-κB pathway, in ex vivo cerebellar slices. RESULTS: Hyperammonemia induced microglial and astrocyte activation at 1 week. This was followed by TNF-a induction in both glial cell types at 2 weeks and in Purkinje neurons at 4 weeks. The level of TNF-a mRNA increased in parallel with the TNF-a protein level, indicating that TNF-a was synthesized in Purkinje cells. This increase was associated with increased NF-κB nuclear translocation. The nuclear translocation of NF-κB and the increase in TNF-a were reversed by R7050, indicating that they were mediated by the activation of TNFR1. Preventing peripheral inflammation with an anti-TNF-a antibody prevents TNF-a induction. CONCLUSION: Sustained (4 weeks) but not short-term hyperammonemia induces TNF-a in Purkinje neurons in rats. This is mediated by peripheral inflammation. TNF-a is also increased in the Purkinje neurons of patients who die with liver cirrhosis. The results suggest that hyperammonemia induces TNF-a in glial cells and that TNF-a released by glial cells activates TNFR1 in Purkinje neurons, leading to NF-κB nuclear translocation and the induction of TNF-a expression, which may contribute to the neurological alterations observed in hyperammonemia and hepatic encephalopathy.

Revisiting the effect of time of birth on neonatal, infant, and child mortality in rural Spain, 1830-1929.

OBJECTIVES: This article aims to determine whether different patterns of mortality occurred among children born during the day and the night respectively, between 1830 and 1929. METHODS: The data include the time of birth and death of 9814 individuals from 10 villages in rural Spain between 1830 and 1929, within a context of natural births at home with little medical support. These data were subjected to a comparative analysis relating to the time of birth and the age at death. RESULTS: Neonatal, infant, and child mortality was higher for children born during daytime. The day-to-night mortality pattern diverged until children were at least 5 years old. CONCLUSIONS: The results confirm that the mortality patterns differed according to the time of birth. Possibly some of these children experienced longer or problematic deliveries that, in the absence of good medical assistance, had health consequences during the following days and years of life.

A Rev-CBP80-eIF4AI complex drives Gag synthesis from the HIV-1 unspliced mRNA.

Gag synthesis from the full-length unspliced mRNA is critical for the production of the viral progeny during human immunodeficiency virus type-1 (HIV-1) replication. While most spliced mRNAs follow the canonical gene expression pathway in which the recruitment of the nuclear cap-binding complex (CBC) and the exon junction complex (EJC) largely stimulates the rates of nuclear export and translation, the unspliced mRNA relies on the viral protein Rev to reach the cytoplasm and recruit the host translational machinery. Here, we confirm that Rev ensures high levels of Gag synthesis by driving nuclear export and translation of the unspliced mRNA. These functions of Rev are supported by the CBC subunit CBP80, which binds Rev and the unspliced mRNA in the nucleus and the cytoplasm. We also demonstrate that Rev interacts with the DEAD-box RNA helicase eIF4AI, which translocates to the nucleus and cooperates with the viral protein to promote Gag synthesis. Finally, we show that the Rev/RRE axis is important for the assembly of a CBP80-eIF4AI complex onto the unspliced mRNA. Together, our results provide further evidence towards the understanding of the molecular mechanisms by which Rev drives Gag synthesis from the unspliced mRNA during HIV-1 replication.

DEAD-box RNA helicase DDX3 connects CRM1-dependent nuclear export and translation of the HIV-1 unspliced mRNA through its N-terminal domain.

DEAD-box RNA helicase DDX3 is a host factor essential for HIV-1 replication and thus, a potential target for novel therapies aimed to overcome viral resistance. Previous studies have shown that DDX3 promotes nuclear export and translation of the HIV-1 unspliced mRNA. Although the function of DDX3 during both processes requires its catalytic activity, it is unknown whether other domains surrounding the helicase core are involved. Here, we show the involvement of the N- and C-terminal domains of DDX3 in the regulation of HIV-1 unspliced mRNA translation. Our results suggest that the intrinsically disordered N-terminal domain of DDX3 regulates its functions in translation by acting prior to the recruitment of the 43S pre-initiation complex onto the viral 5'-UTR. Interestingly, this regulation was conserved in HIV-2 and was dependent on the CRM1-dependent nuclear export pathway suggesting a role of the RNA helicase in interconnecting nuclear export with ribosome recruitment of the viral unspliced mRNA. This specific function of DDX3 during HIV gene expression could be exploited as an alternative target for pharmaceutical intervention.

HIV-2 genomic RNA accumulates in stress granules in the absence of active translation.

During the post-transcriptional events of the HIV-2 replication cycle, the full-length unspliced genomic RNA (gRNA) is first used as an mRNA to synthesize Gag and Gag-Pol proteins and then packaged into progeny virions. However, the mechanisms responsible for the coordinate usage of the gRNA during these two mutually exclusive events are poorly understood. Here, we present evidence showing that HIV-2 expression induces stress granule assembly in cultured cells. This contrasts with HIV-1, which interferes with stress granules assembly even upon induced cellular stress. Moreover, we observed that the RNA-binding protein and stress granules assembly factor TIAR associates with the gRNA to form a TIAR-HIV-2 ribonucleoprotein (TH2RNP) complex localizing diffuse in the cytoplasm or aggregated in stress granules. Although the assembly of TH2RNP in stress granules did not require the binding of the Gag protein to the gRNA, we observed that increased levels of Gag promoted both translational arrest and stress granule assembly. Moreover, HIV-2 Gag also localizes to stress granules in the absence of a 'packageable' gRNA. Our results indicate that the HIV-2 gRNA is compartmentalized in stress granules in the absence of active translation prior to being selected for packaging by the Gag polyprotein.

Strengthening safety compliance in nuclear power operations: a role-based approach.

Safety compliance is of paramount importance in guaranteeing the safe running of nuclear power plants. However, it depends mostly on procedures that do not always involve the safest outcomes. This article introduces an empirical model based on the organizational role theory to analyze the influence of legitimate sources of expectations (procedures formalization and leadership) on workers' compliance behaviors. The sample was composed of 495 employees from two Spanish nuclear power plants. Structural equation analysis showed that, in spite of some problematic effects of proceduralization (such as role conflict and role ambiguity), procedure formalization along with an empowering leadership style lead to safety compliance by clarifying a worker's role in safety. Implications of these findings for safety research are outlined, as well as their practical implications.

Analyzing the effects of the Spanish Civil War on biological well-being through new anthropometric indicators.

Numerous studies have demonstrated the negative impact of severe economic shocks (such as those associated with wars) on the growth of children and adolescents. Individuals exposed to these shocks during their developmental years exhibited shorter average heights compared to both previous and subsequent generations. Anthropometric research has highlighted the sensitivity of the height variable in understanding the biological well-being of children and adolescents. However, little attention has been paid to the evolution of other anthropometric variables. This study investigates the impact of the famine following the Spanish Civil War on biological well-being in nine municipalities of the Region of Valencia (with over 120,000 observations of individuals born between 1890 and 1955) using two indicators: chest circumference, and the percentage of individuals of short stature. Our findings confirm that both of these anthropometric indicators were responsive to the economic shock of the Civil War. The well-being levels prior to the war took 20 years to recover.

Height and political activism in rural Aragón (Spain) during the 20th century. A new perspective using individual-level data.

This article explores the relationship between the political leanings of more than 1000 men born in the 1870-1970 s in 11 rural Aragonese villages and their biological well-being during childhood and adolescence, proxied by height. The aim is to test whether an individual was more likely to be left-wing if his level of biological well-being was lower and, therefore, with more incentives to fight against the social inequality that had negatively affected his family. Our results confirm that, for most of the study period, there was a strong relationship between shorter height and political activism1 in left-wing parties and organizations.

Microenvironmental Snail1-induced immunosuppression promotes melanoma growth.

Melanoma is an aggressive form of skin cancer due to its high metastatic abilities and resistance to therapies. Melanoma cells reside in a heterogeneous tumour microenvironment that acts as a crucial regulator of its progression. Snail1 is an epithelial-to-mesenchymal transition transcription factor expressed during development and reactivated in pathological situations including fibrosis and cancer. In this work, we show that Snail1 is activated in the melanoma microenvironment, particularly in fibroblasts. Analysis of mouse models that allow stromal Snail1 depletion and therapeutic Snail1 blockade indicate that targeting Snail1 in the tumour microenvironment decreases melanoma growth and lung metastatic burden, extending mice survival. Transcriptomic analysis of melanoma-associated fibroblasts and analysis of the tumours indicate that stromal Snail1 induces melanoma growth by promoting an immunosuppressive microenvironment and a decrease in anti-tumour immunity. This study unveils a novel role of Snail1 in melanoma biology and supports its potential as a therapeutic target.

Selective NO2 Detection of CaCu3Ti4O12 Ceramic Prepared by the Sol-Gel Technique and DRIFT Measurements to Elucidate the Gas Sensing Mechanism.

NO2 is one of the main greenhouse gases, which is mainly generated by the combustion of fossil fuels. In addition to its contribution to global warming, this gas is also directly dangerous to humans. The present work reports the structural and gas sensing properties of the CaCu3Ti4O12 compound prepared by the sol-gel technique. Rietveld refinement confirmed the formation of the pseudo-cubic CaCu3Ti4O12 compound, with less than 4 wt% of the secondary phases. The microstructural and elemental composition analysis were carried out using scanning electron microscopy and X-ray energy dispersive spectroscopy, respectively, while the elemental oxidation states of the samples were determined by X-ray photoelectron spectroscopy. The gas sensing response of the samples was performed for different concentrations of NO2, H2, CO, C2H2 and C2H4 at temperatures between 100 and 300 °C. The materials exhibited selectivity for NO2, showing a greater sensor signal at 250 °C, which was correlated with the highest concentration of nitrite and nitrate species on the CCTO surface using DRIFT spectroscopy.

Nanostructured manganese oxide particles from coordination complex decomposition and their catalytic properties for ethanol oxidation.

Novel manganese oxide particles with complex morphologies and different nanostructures (i.e., spherical/lamellar) were synthesized by initial preparation of a coordination complex of manganese with 1,4,7,10-tetraazacyclododecane (cyclen), followed by characterization of the nanostructured oxide as a catalytic material for ethanol oxidation. The samples present a bulk gamma-MnO2 structure although X-ray photoelectron spectroscopy analysis reveals that their surfaces have different chemical compositions. Some of these nanostructured particles show high catalytic activities for ethanol oxidation enabling a decrease of the reaction temperature by more than 80 degrees C as compared with traditional MnO2 particles. The high catalytic activity of the particles depends on their morphology and a relationship between morphology and specific area was established. It is proposed that these novel nanostructured manganese oxide particles may be highly active in the catalytic oxidation of other volatile organic compounds (VOCs) opening up their further development for environmental applications.

Bidirectional genome-wide CRISPR screens reveal host factors regulating SARS-CoV-2, MERS-CoV and seasonal HCoVs.

CRISPR knockout (KO) screens have identified host factors regulating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication. Here, we conducted a meta-analysis of these screens, which showed a high level of cell-type specificity of the identified hits, highlighting the necessity of additional models to uncover the full landscape of host factors. Thus, we performed genome-wide KO and activation screens in Calu-3 lung cells and KO screens in Caco-2 colorectal cells, followed by secondary screens in four human cell lines. This revealed host-dependency factors, including AP1G1 adaptin and ATP8B1 flippase, as well as inhibitors, including mucins. Interestingly, some of the identified genes also modulate Middle East respiratory syndrome coronavirus (MERS-CoV) and seasonal human coronavirus (HCoV) (HCoV-NL63 and HCoV-229E) replication. Moreover, most genes had an impact on viral entry, with AP1G1 likely regulating TMPRSS2 activity at the plasma membrane. These results demonstrate the value of multiple cell models and perturbational modalities for understanding SARS-CoV-2 replication and provide a list of potential targets for therapeutic interventions.

The association between male height and lifespan in rural Spain, birth cohorts 1835-1939.

This article analyses the relationship between male height and age at death and its evolution over time among conscripts born in fourteen villages in north-east rural Spain. A total of 1,488 conscripts born between 1835 and 1939 (and who died between 1868 and 2019) have been included in the analysis (based on the study of 3 sub-periods: 1835-1869, 1870-1899, and 1900-1939). The height data have been obtained from military service conscription records and the demographic and socioeconomic information of the deceased was obtained from parish archives and censuses. The data have been linked according to nominative criteria using family reconstitution methods. For the statistical analysis, we have used ordinary least squares (OLS) linear regressions with heteroskedasticity-robust estimation. The results suggest a positive relationship between height and lifespan in the long-term. For the birth cohorts of 1835-1869, conscripts with a height of 170 cm or more lived on average 7.6 years longer than conscripts measuring less than 160 cm. This difference in life expectancy tended to disappear for the birth cohorts of 1900-1939, benefiting especially the short conscripts who had greater possibilities to increase their average lifespan. The reasons that might explain these changes could reside in the improvements experienced by this group in terms of their living conditions, health and nutrition during the twentieth century.

Rethinking the Fertility Transition in Rural Aragón (Spain) Using Height Data.

Based on an analysis of the life trajectories of 2510 conscripts and their families from a Spanish rural area in the period 1835-1977, this paper studies the development of the fertility transition in relation to height using bivariate analyses. The use of heights is an innovative perspective of delving into the fertility transition and social transformation entailed. The results confirm that the men with a low level of biological well-being (related to low socio-economic groups) were those who started to control their fertility, perhaps due to the effect that increased average family size had on their budget. The children of individuals who controlled their fertility were taller than the children of other families. Therefore, the children of parents who controlled their fertility experienced the largest intergenerational increase in height (approximately 50% higher). This increase could be due to the consequence of a greater investment in children (Becker's hypothesis) or a greater availability of resources for the whole family (resource dilution hypothesis).

Did parental care in early life affect height? Evidence from rural Spain (19th-20th centuries).

This article examines the relationship between childhood mortality experienced within families and the height of surviving male children. Sibling mortality, controlled by different socioeconomic and environmental variables, is used as an approximation of the hygienic and epidemiological context and practices within the family. The analysis is based on a sample of 2783 individuals born between 1835 and 1977 in 14 villages in north-eastern Spain. The mortality data were obtained from the parish archives of the reference villages, and the height data from military service records of conscriptions at 21 years of age. The data were linked according to nominative criteria using family reconstitution methods. The results suggest the existence of a strong negative relationship between height and the childhood mortality experienced within families. Children born in families in which 50% of the children died before the age of five were up to 2.3 cm shorter than those of families with childhood mortality of less than 25%. General socioeconomic, hygienic and health improvements reduced childhood mortality, causing this link to gradually disappear between the 1940s and 1970s.

Bidirectional genome-wide CRISPR screens reveal host factors regulating SARS-CoV-2, MERS-CoV and seasonal coronaviruses.

Several genome-wide CRISPR knockout screens have been conducted to identify host factors regulating SARS-CoV-2 replication, but the models used have often relied on overexpression of ACE2 receptor. Additionally, such screens have yet to identify the protease TMPRSS2, known to be important for viral entry at the plasma membrane. Here, we conducted a meta-analysis of these screens and showed a high level of cell-type specificity of the identified hits, arguing for the necessity of additional models to uncover the full landscape of SARS-CoV-2 host factors. We performed genome-wide knockout and activation CRISPR screens in Calu-3 lung epithelial cells, as well as knockout screens in Caco-2 intestinal cells. In addition to identifying ACE2 and TMPRSS2 as top hits, our study reveals a series of so far unidentified and critical host-dependency factors, including the Adaptins AP1G1 and AP1B1 and the flippase ATP8B1. Moreover, new anti-SARS-CoV-2 proteins with potent activity, including several membrane-associated Mucins, IL6R, and CD44 were identified. We further observed that these genes mostly acted at the critical step of viral entry, with the notable exception of ATP8B1, the knockout of which prevented late stages of viral replication. Exploring the pro- and anti-viral breadth of these genes using highly pathogenic MERS-CoV, seasonal HCoV-NL63 and -229E and influenza A orthomyxovirus, we reveal that some genes such as AP1G1 and ATP8B1 are general coronavirus cofactors. In contrast, Mucins recapitulated their known role as a general antiviral defense mechanism. These results demonstrate the value of considering multiple cell models and perturbational modalities for understanding SARS-CoV-2 replication and provide a list of potential new targets for therapeutic interventions.