SRRM2 splicing factor modulates cell fate in early development.

Embryo development is an orchestrated process that relies on tight regulation of gene expression to guide cell differentiation and fate decisions. The Srrm2 splicing factor has recently been implicated in developmental disorders and diseases, but its role in early mammalian development remains unexplored. Here, we show that Srrm2 dosage is critical for maintaining embryonic stem cell pluripotency and cell identity. Srrm2 heterozygosity promotes loss of stemness, characterised by the coexistence of cells expressing naive and formative pluripotency markers, together with extensive changes in gene expression, including genes regulated by serum-response transcription factor (SRF) and differentiation-related genes. Depletion of Srrm2 by RNA interference in embryonic stem cells shows that the earliest effects of Srrm2 heterozygosity are specific alternative splicing events on a small number of genes, followed by expression changes in metabolism and differentiation-related genes. Our findings unveil molecular and cellular roles of Srrm2 in stemness and lineage commitment, shedding light on the roles of splicing regulators in early embryogenesis, developmental diseases and tumorigenesis.

Targeting SRSF2 mutations in leukemia with RKI-1447: A strategy to impair cellular division and nuclear structure.

Spliceosome machinery mutations are common early mutations in myeloid malignancies; however, effective targeted therapies against them are still lacking. In the current study, we used an in vitro high-throughput drug screen among four different isogenic cell lines and identified RKI-1447, a Rho-associated protein kinase inhibitor, as selective cytotoxic effector of SRSF2 mutant cells. RKI-1447 targeted SRSF2 mutated primary human samples in xenografts models. RKI-1447 induced mitotic catastrophe and induced major reorganization of the microtubule system and severe nuclear deformation. Transmission electron microscopy and 3D light microscopy revealed that SRSF2 mutations induce deep nuclear indentation and segmentation that are apparently driven by microtubule-rich cytoplasmic intrusions, which are exacerbated by RKI-1447. The severe nuclear deformation in RKI-1447-treated SRSF2 mutant cells prevents cells from completing mitosis. These findings shed new light on the interplay between microtubules and the nucleus and offers new ways for targeting pre-leukemic SRSF2 mutant cells.

Mpox outbreak in Rio de Janeiro, Brazil: A translational approach.

Mpox is a zoonotic disease historically reported in Africa. Since 2003, limited outbreaks have occurred outside Africa. In 2022, the global spread of cases with sustained interhuman transmission and unusual disease features raised public health concerns. We explore the mpox outbreak in Rio de Janeiro (RJ) state, Brazil, in an observational study of mpox-suspected cases from June to December 2022. Data collection relied on a public healthcare notification form. Diagnosis was determined by MPXV-PCR. In 46 confirmed cases, anti-OPXV IgG was determined by ELISA, and seven MPXV genomes were sequenced. A total of 3095 cases were included, 816 (26.3%) with positive MPXV-PCR results. Most positive cases were men in their 30 s and MSM. A total of 285 (34.9%) MPXV-PCR+ patients live with HIV. Eight were coinfected with varicella-zoster virus. Anogenital lesions and adenomegaly were associated with the diagnosis of mpox. Females and individuals under 18 represented 9.4% and 5.4% of all confirmed cases, respectively, showing higher PCR cycle threshold (Ct) values and fewer anogenital lesions compared to adult men. Anti-OPXV IgG was detected in 29/46 (63.0%) patients. All analyzed sequences belonged to clade IIb. In RJ state, mpox presented a diverse clinical picture, represented mainly by mild cases with low complication rates and prominent genital involvement. The incidence in females and children was higher than usually reported. The observation of a bimodal distribution of Ct values, with few positive results, may suggest the need to review the diagnostic criteria in these groups.

A Versatile Shaping Method of Very-High Loading Porous Solids Paper Adsorbent Composites.

Owing to their high porosity and tunability, porous solids such as metal-organic frameworks (MOFs), zeolites, or activated carbons (ACs) are of great interest in the fields of air purification, gas separation, and catalysis, among others. Nonetheless, these materials are usually synthetized as powders and need to be shaped in a more practical way that does not modify their intrinsic property (i.e., porosity). Elaborating porous, freestanding and flexible sheets is a relevant shaping strategy. However, when high loadings (>70 wt.%) are achieved the mechanical properties are challenged. A new straightforward and green method involving the combination softwood bleached kraft pulp fibers (S) and nano-fibrillated cellulose (NFC) is reported, where S provides flexibility while NFC acts as a micro-structuring and mechanical reinforcement agent to form high loadings porous solids paper sheets (>70 wt.%). The composite has unobstructed porosity and good mechanical strength. The sheets prepared with various fillers (MOFs, ACs, and zeolites) can be rolled, handled, and adapted to different uses, such as air purification. As an example of potential application, a MOF paper composite has been considered for the capture of polar volatile organic compounds exhibiting better performance than beads and granules.

Characteristics of women diagnosed with mpox infection compared to men: A case series from Brazil.

BACKGROUND: Cisgender men were mostly affected during the 2022 mpox multinational outbreak, with few cases reported in women. This study compares the characteristics of individuals diagnosed with mpox infection according to gender in Rio de Janeiro. METHODS: We obtained surveillance data of mpox cases notified to Rio de Janeiro State Health Department (June 12 to December 15, 2022). We compared women (cisgender or transgender) to men (cisgender or transgender) using chi-squared, Fisher's exact, and Mood's median tests. RESULTS: A total of 1306 mpox cases were reported; 1188 (91.0%) men (99.8% cisgender, 0.2% transgender), 108 (8.3%) women (87.0% cisgender, 13.0% transgender), and 10 (0.8%) non-binary persons. Compared to men, women were more frequently older (40+years: 34.3% vs. 25.1%; p < 0.001), reported more frequent non-sexual contact with a potential mpox case (21.4% vs. 9.8%; p = 0.004), fewer sexual partnerships (10.9 vs. 54.8%; p < 0.001), less sexual contact with a potential mpox case (18.5% vs. 43.0%; p < 0.001), fewer genital lesions (31.8% vs. 57.9%; p < 0.001), fewer systemic mpox signs/symptoms (38.0% vs. 50.1%; p = 0.015) and had a lower HIV prevalence (8.3% vs. 46.3%; p < 0.001), with all cases among transgender women. Eight women were hospitalized; no deaths occurred. The highest number of cases among women were notified in epidemiological week 34, when the number of cases among men started to decrease. CONCLUSIONS: Women diagnosed with mpox presented differences in epidemiological, behavioral, and clinical characteristics compared to men. Health services should provide a comprehensive assessment that accounts for gender diversity.

Combined Oxidative Phosphorylation Deficiency Type-13 with Perinatal Presentation: A Case Report.

INTRODUCTION: Polynucleotide phosphorylase is involved in RNA processing in mitochondria. Biallelic variants in PNPT1 cause mitochondrial RNA import protein deficiency and heterogeneous clinical manifestations. CASE REPORT: The patiest was the first child of remote consanguineous parents, born at 35 weeks by caesarean section due to fetal growth restriction. Apgar index was 9/10/10. Birth weight, length and head circumference were at 3rd, <3rd and 10th percentiles, respectively. In the first hours of life, respiratory distress, hypoglycaemia and seizures ensued. She started invasive mechanic ventilation, phenobarbital and was transferred to ICU. Physical examination showed minor facial dysmorphisms, brief eye-opening, hypotonia and hyporeflexia. Electroencephalogram showed immature pattern and multifocal paroxysmal activity. MRI at D8 of life showed severe reduced brain volume. Normal aminoacid screen was also observed. Expanded newborn screening was negative. Mitochondrial organic aciduria was seen. WES showed a homozygotic likely pathogenic variant in the PNPT1 gene. MRI at 6-months showed brain atrophy, thin corpus callosum, reduced brainstem volume. Bilateral and symmetrical lesions in globi pallidi, compatible with Leigh síndrome were observed. Currently, at 14 months, no neurodevelopment progress, dystonia, visual deficit, sensorineural deafness, hypertrophic cardiomyopathy and microcephaly are observed. CONCLUSION: The early and severe Leigh-like presentation of our patient expands the phenotype spectrum of this disease. As far as we know, this is the first reported case of PNPT1 mutation with onset in the perinatal period. Moreover, hypertrophic cardiomyopathy has not yet been described in association with mutation of the PNPT1 gene. WES was the key for early diagnosis in this patient. It should be done in all children with severe clinical presentation of unknown origin.

Specific quinone reductase 2 inhibitors reduce metabolic burden and reverse Alzheimer's disease phenotype in mice.

Biological aging can be described as accumulative, prolonged metabolic stress and is the major risk factor for cognitive decline and Alzheimer's disease (AD). Recently, we identified and described a quinone reductase 2 (QR2) pathway in the brain, in which QR2 acts as a removable memory constraint and metabolic buffer within neurons. QR2 becomes overexpressed with age, and it is possibly a novel contributing factor to age-related metabolic stress and cognitive deficit. We found that, in human cells, genetic removal of QR2 produced a shift in the proteome opposing that found in AD brains while simultaneously reducing oxidative stress. We therefore created highly specific QR2 inhibitors (QR2is) to enable evaluation of chronic QR2 inhibition as a means to reduce biological age-related metabolic stress and cognitive decline. QR2is replicated results obtained by genetic removal of QR2, while local QR2i microinjection improved hippocampal and cortical-dependent learning in rats and mice. Continuous consumption of QR2is in drinking water improved cognition and reduced pathology in the brains of AD-model mice (5xFAD), with a noticeable between-sex effect on treatment duration. These results demonstrate the importance of QR2 activity and pathway function in the healthy and neurodegenerative brain and what we believe to be the great therapeutic potential of QR2is as first-in-class drugs.

Maldi-Tof-Ms Identified Rhodococcus Hoagie Bacteraemia In An Immunocompromised Patient.

Rhodococcus hoagie, previously referred to as R. equi, is a Gram-positive intracellular coccobacillus that belongs to the Nocardiaceae family. This multi-host pathogen causes infections in farm animals, particularly foals, but also in immunosuppressed patients, mainly individuals treated with high doses of corticosteroids, subjected to organ transplant, or infected with human immunodeficiency virus Objectives of the study are to report a bloodstream infection in an immunocompromised patient. Immunocompromised patients with advanced HIV who presented bloodstream infection, residing in an urban setting and having undertaken no trips to the countryside or elsewhere during the COVID-19 pandemic. Blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was done in order to identify the bacteria. The immunocompromised female patient presented bloodstream infection by Rhodococcus hoagie, which was identified using MALDI-TOF-MS. R. hoagie can cause a severe infection with a high mortality rate if prompt treatment with a combination of antibiotics is not established. A high level of suspicion is required to establish the diagnosis, as it may be misdiagnosed as pulmonary tuberculosis. On gram stain, R. hoagie may appear as beaded to solid staining coccobacilli, which can be dismissed as a "diphtheroid" contaminant. The infection was identified using MALDI-TOF-MS.

Spatio-Temporal Cluster Detection of Dengue, Chikungunya, and Zika Viruses' Infection in Rio de Janeiro State from 2010 to 2019.

Dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV) virus infections are widespread throughout the Rio de Janeiro state. The co-circulation of these emergent arboviruses constitutes a serious public health problem, resulting in outbreaks that can spatially and temporally overlap. Environmental conditions favor the presence, maintenance, and expansion of Aedes aegypti, the primary vector of these urban arboviruses. This study assessed the detection of clusters of urban arboviruses in the Rio de Janeiro state from 2010 to 2019. Notified cases of dengue, chikungunya, and Zika were grouped by year according to the onset of symptoms and their municipality of residence. The study period recorded the highest number of dengue epidemics in the state along with the simultaneous circulation of chikungunya and Zika viruses. The analyzes showed that the central municipalities of the metropolitan regions were associated with higher risk areas. Central municipalities in metropolitan regions were the first most likely clusters for dengue and Zika, and the second most likely cluster for chikungunya. Furthermore, the northwest and north regions were comprised clusters with the highest relative risk for the three arboviruses, underscoring the impact of these arboviruses in less densely populated regions of Brazil. The identification of high-risk areas over time highlights the need for effective control measures, targeted prevention and control interventions for these urban arboviral diseases.

Assessing the climate change adaptability of sustainable land management practices regarding water availability and quality: A case study in the Sorraia catchment, Portugal.

In Mediterranean catchments, such as the Sorraia catchment in Portugal, it is expected that climate change will increase drought stress and the deterioration of water quality in reservoirs. Sustainable land management (SLM) practices are seen as an adaption measure for those problems, but the effectiveness on improving climate change impacted water availability and quality on catchment scale is still poorly understood. Therefore, this study aims to evaluate the effectiveness of SLM practices in adapting the impacts of climate change on water availability and quality of the Montargil and Maranhão reservoirs in the Sorraia catchment. A well-calibrated Soil Water Assessment Tool model is used to simulate four scenarios (2041-2071 and 2071-2100; representative climate pathways 4.5 and 8.5), to investigate the effects of climate change on total phosphorus load (TP) in streams, reservoir volume, irrigation use and water exploitation index (WEI). Results showed that WEI will not exceed any water stress level while reservoir water quality will worsen. In particular since the TP load in streams flowing into the reservoirs increases and the volume decreases, it is likely that the existing P limitation for eutrophication will be counteracted. Nevertheless, tested SLM practices were able to decrease the TP load in those streams and increase the reservoir volume under future climates. Overall, this study shows that the SLM practices are effective in adapting to the climate change effects regarding reservoir water quality, without worsening the water availability; thus, it is a promising tool that should be investigated further for application by e.g. local land-users and decision makers.

Identifying a selective inhibitor of autophagy that targets ATG12-ATG3 protein-protein interaction.

Macroautophagy/autophagy is a catabolic process by which cytosolic content is engulfed, degraded and recycled. It has been implicated as a critical pathway in advanced stages of cancer, as it maintains tumor cell homeostasis and continuous growth by nourishing hypoxic or nutrient-starved tumors. Autophagy also supports alternative cellular trafficking pathways, providing a mechanism of non-canonical secretion of inflammatory cytokines. This opens a significant therapeutic opportunity for using autophagy inhibitors in cancer and acute inflammatory responses. Here we developed a high throughput compound screen to identify inhibitors of protein-protein interaction (PPI) in autophagy, based on the protein-fragment complementation assay (PCA). We chose to target the ATG12-ATG3 PPI, as this interaction is indispensable for autophagosome formation, and the analyzed structure of the interaction interface predicts that it may be amenable to inhibition by small molecules. We screened 41,161 compounds yielding 17 compounds that effectively inhibit the ATG12-ATG3 interaction in the PCA platform, and which were subsequently filtered by their ability to inhibit autophagosome formation in viable cells. We describe a lead compound (#189) that inhibited GFP-fused MAP1LC3B/LC3B (microtubule associated protein 1 light chain 3 beta) puncta formation in cells with IC50 value corresponding to 9.3 µM. This compound displayed a selective inhibitory effect on the growth of autophagy addicted tumor cells and inhibited secretion of IL1B/IL-1ß (interleukin 1 beta) by macrophage-like cells. Compound 189 has the potential to be developed into a therapeutic drug and its discovery documents the power of targeting PPIs for acquiring specific and selective compound inhibitors of autophagy.Abbreviations: ANOVA: analysis of variance; ATG: autophagy related; CQ: chloroquine; GFP: green fluorescent protein; GLuc: Gaussia Luciferase; HEK: human embryonic kidney; IL1B: interleukin 1 beta; LPS: lipopolysaccharide; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; PCA: protein-fragment complementation assay; PDAC: pancreatic ductal adenocarcinoma; PMA: phorbol 12-myristate 13-acetate; PPI: protein-protein interaction. VCL: vinculin.

Evaluation of an Imine-Linked Polymer Organic Framework for Storage and Release of H2S and NO.

Hydrogen sulfide (H2S) and nitric oxide (NO) are especially known as toxic and polluting gases, yet they are also endogenously produced and play key roles in numerous biological processes. These two opposing aspects of the gases highlight the need for new types of materials to be developed in addition to the most common materials such as activated carbons and zeolites. Herein, a new imine-linked polymer organic framework was obtained using the inexpensive and easy-to-access reagents isophthalaldehyde and 2,4,6-triaminopyrimidine in good yield (64%) through the simple and catalyst-free Schiff-base reaction. The polymeric material has microporosity, an ABET surface area of 51 m2/g, and temperature stability up to 300 °C. The obtained 2,4,6-triaminopyrimidine imine-linked polymer organic material has a higher capacity to adsorb NO (1.6 mmol/g) than H2S (0.97 mmol/g). Release studies in aqueous solution showed that H2S has a faster release (3 h) from the material than NO, for which a steady release was observed for at least 5 h. This result is the first evaluation of the possibility of an imine-linked polymer organic framework being used in the therapeutic release of NO or H2S.

Environmental and topographic drivers of amphibian phylogenetic diversity and endemism in the Iberian Peninsula.

Understanding the ecological and evolutionary processes driving biodiversity patterns and allowing their persistence is of utmost importance. Many hypotheses have been proposed to explain spatial diversity patterns, including water-energy availability, habitat heterogeneity, and historical climatic refugia. The main goal of this study is to identify if general spatial drivers of species diversity patterns of phylogenetic diversity (PD) and phylogenetic endemism (PE) at the global scale are also predictive of PD and PE at regional scales, using Iberian amphibians as a case study. Our main hypothesis assumes that topography along with contemporary and historical climate are drivers of phylogenetic diversity and endemism, but that the strength of these predictors may be weaker at the regional scale than it tends to be at the global scale. We mapped spatial patterns of Iberian amphibians' phylogenetic diversity and endemism, using previously published phylogenetic and distribution data. Furthermore, we compiled spatial data on topographic and climatic variables related to the water-energy availability, topography, and historical climatic instability hypotheses. To test our hypotheses, we used Spatial Autoregressive Models and selected the best model to explain diversity patterns based on Akaike Information Criterion. Our results show that, out of the variables tested in our study, water-energy availability and historical climate instability are the most important drivers of amphibian diversity in Iberia. However, as predicted, the strength of these predictors in our case study is weaker than it tends to be at global scales. Thus, additional drivers should also be investigated and we suggest caution when interpreting these predictors as surrogates for different components of diversity.

Maximising signal-to-noise ratios in environmental DNA-based monitoring.

Man's impacts on global ecosystems are increasing and there is a growing demand that these activities be appropriately monitored. Monitoring requires measurement of a response metric ('signal') that changes maximally and consistently in response to the monitored activity irrespective of other factors ('noise'), thus maximising the signal-to-noise ratio. Indices derived from time-consuming morphology-based taxonomic identification of organisms are a core part of many monitoring programmes. Metabarcoding is an alternative to morphology-based identification and involves the sequencing of short fragments of DNA ('markers') from multiple taxa simultaneously. DNA suitable for metabarcoding includes that extracted from environmental samples (eDNA). Metabarcoding outputs DNA sequences that can be identified (annotated) by matching them against archived annotated sequences. However, sequences from most organisms are not archived - preventing annotation and potentially limiting metabarcoding in monitoring applications. Consequently, there is growing interest in using unannotated sequences as response metrics in monitoring programmes. We compared the sequences from three commonly used markers (16S (V3/V4 regions), 18S (V1/V2 regions) and COI) and, sampling along steep impact gradients, showed that the 16S and COI sequences were associated with the largest and smallest signal-to-noise ratio respectively. We trialled four separate, intuitive, noise-reduction approaches and demonstrated that removing less frequent sequences improved the signal-to-noise ratio, partitioning an additional 25 % from noise to explanatory factors in non-parametric ANOVA (NPA) and reducing dispersion in the data. For the 16S marker, retaining only the most frequently observed sequence, per sample, resulting in nine sequences across 150 samples, generated a near-maximal signal-to-noise ratio (95 % of the variance explained in NPA). We recommend that NPA, combined with rigorous elimination of less frequent sequences, be used to pre-filter sequences/taxa being used in monitoring applications. Our approach will simplify downstream analysis, for example the identification of key taxa and functional associations.

Molecular ecology meets systematic conservation planning.

Integrative and proactive conservation approaches are critical to the long-term persistence of biodiversity. Molecular data can provide important information on evolutionary processes necessary for conserving multiple levels of biodiversity (genes, populations, species, and ecosystems). However, molecular data are rarely used to guide spatial conservation decision-making. Here, we bridge the fields of molecular ecology (ME) and systematic conservation planning (SCP) (the 'why') to build a foundation for the inclusion of molecular data into spatial conservation planning tools (the 'how'), and provide a practical guide for implementing this integrative approach for both conservation planners and molecular ecologists. The proposed framework enhances interdisciplinary capacity, which is crucial to achieving the ambitious global conservation goals envisioned for the next decade.

DNA damage independent inhibition of NF-κB transcription by anthracyclines.

Anthracyclines are among the most used and effective anticancer drugs. Their activity has been attributed to DNA double-strand breaks resulting from topoisomerase II poisoning and to eviction of histones from select sites in the genome. Here, we show that the extensively used anthracyclines Doxorubicin, Daunorubicin, and Epirubicin decrease the transcription of nuclear factor kappa B (NF-κB)-dependent gene targets, but not interferon-responsive genes in primary mouse (Mus musculus) macrophages. Using an NMR-based structural approach, we demonstrate that anthracyclines disturb the complexes formed between the NF-κB subunit RelA and its DNA-binding sites. The anthracycline variants Aclarubicin, Doxorubicinone, and the newly developed Dimethyl-doxorubicin, which share anticancer properties with the other anthracyclines but do not induce DNA damage, also suppressed inflammation, thus uncoupling DNA damage from the effects on inflammation. These findings have implications for anticancer therapy and for the development of novel anti-inflammatory drugs with limited side effects for life-threatening conditions such as sepsis.

Phylodynamic analysis of SARS-CoV-2 spread in Rio de Janeiro, Brazil, highlights how metropolitan areas act as dispersal hubs for new variants.

During the first semester of 2021, all of Brazil has suffered an intense wave of COVID-19 associated with the Gamma variant. In July, the first cases of Delta variant were detected in the state of Rio de Janeiro. In this work, we have employed phylodynamic methods to analyse more than 1 600 genomic sequences of Delta variant collected until September in Rio de Janeiro to reconstruct how this variant has surpassed Gamma and dispersed throughout the state. After the introduction of Delta, it has initially spread mostly in the homonymous city of Rio de Janeiro, the most populous of the state. In a second stage, dispersal occurred to mid- and long-range cities, which acted as new close-range hubs for spread. We observed that the substitution of Gamma by Delta was possibly caused by its higher viral load, a proxy for transmissibility. This variant turnover prompted a new surge in cases, but with lower lethality than was observed during the peak caused by Gamma. We reason that high vaccination rates in the state of Rio de Janeiro were possibly what prevented a higher number of deaths.

Cryptosporidium diagnosis in different groups of children and characterization of parasite species.

BACKGROUND: Microscopy and enzyme-linked immunosorbent assay (ELISA) are routinely used for Cryptosporidium diagnosis, without differentiating the parasite species. METHODS: Children's feces were analyzed by modified Ziehl-Neelsen (mZN) and ELISA for Cryptosporidium diagnosis and by polymerase chain reaction-restriction fragment length polymorphism for species identification. RESULTS: Cryptosporidium frequency was 2.6%. The sensitivity and specificity of ELISA were 85.7% and 99.7%, respectively, with excellent concordance with mZN (kappa=0.854). Parasite species were characterized as Cryptosporidium hominis (78.3%), Cryptosporidium felis (17.4%), and Cryptosporidium parvum (4.3%). CONCLUSIONS: Coproantigen ELISA is as efficient as mZN for Cryptosporidium diagnosis. Cryptosporidium genotyping suggests anthroponotic and zoonotic transmission to children.

Emergence of Within-Host SARS-CoV-2 Recombinant Genome After Coinfection by Gamma and Delta Variants: A Case Report.

In this study, we report the first case of intra-host SARS-CoV-2 recombination during a coinfection by the variants of concern (VOC) AY.33 (Delta) and P.1 (Gamma) supported by sequencing reads harboring a mosaic of lineage-defining mutations. By using next-generation sequencing reads intersecting regions that simultaneously overlap lineage-defining mutations from Gamma and Delta, we were able to identify a total of six recombinant regions across the SARS-CoV-2 genome within a sample. Four of them mapped in the spike gene and two in the nucleocapsid gene. We detected mosaic reads harboring a combination of lineage-defining mutations from each VOC. To our knowledge, this is the first report of intra-host RNA-RNA recombination between two lineages of SARS-CoV-2, which can represent a threat to public health management during the COVID-19 pandemic due to the possibility of the emergence of viruses with recombinant phenotypes.