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Timed feeding drives adipose browning, although the integrative mechanisms for the same remain unclear. Here, we show that twice-a-night (TAN) feeding generates biphasic oscillations of circulating insulin and leptin, representing their entrainment by timed feeding. Insulin and leptin surges lead to marked cellular, functional, and metabolic remodeling of subcutaneous white adipose tissue (sWAT), resulting in increased energy expenditure. Single-cell RNA-sequencing (scRNA-seq) analyses and flow cytometry demonstrate a role for insulin and leptin surges in innate lymphoid type 2 (ILC2) cell recruitment and sWAT browning, since sWAT depot denervation or loss of leptin or insulin receptor signaling or ILC2 recruitment each dampens TAN feeding-induced sWAT remodeling and energy expenditure. Consistently, recreating insulin and leptin oscillations via once-a-day timed co-injections is sufficient to favorably remodel innervated sWAT. Innervation is necessary for sWAT remodeling, since denervation of sWAT, but not brown adipose tissue (BAT), blocks TAN-induced sWAT remodeling and resolution of inflammation. In sum, reorganization of nutrient-sensitive pathways remodels sWAT and drives the metabolic benefits of timed feeding.

Abstract DNA double-strand breaks (DSBs) must be repaired to ensure cell survival and genomic integrity. In yeast, the Mre11-Rad50-Xrs2 complex (MRX) collaborates with Sae2 to initiate DSB repair. Sae2 stimulates two MRX nuclease activities, endonuclease and 3’−5’ exonuclease. However, how Sae2 controls the two nuclease activities remains enigmatic. Using a combined genetic and biochemical approach, we identified a separation-of-function rad50 mutation, rad50-C47, that causes a defect in Sae2-dependent MRX 3’−5’ exonuclease activity, but not endonuclease activity. We found that both the endo- and 3’−5’ exonuclease activities are essential to release Spo11 from DNA ends, whereas only the endonuclease activity is required for hairpin removal. We also uncovered that MRX-Sae2 endonuclease introduces a cleavage at defined distances from the Spo11-blocked end with gradually decreasing efficiency. Our findings demonstrate that Sae2 stimulates the MRX endo- and exonuclease activities via Rad50 by different mechanisms, ensuring diverse actions of MRX-Sae2 nuclease at DNA ends.

Abstract Aims We aimed to analyse the characteristics and in‐hospital outcomes of patients hospitalized for heart failure (HF) with co‐morbid systemic sclerosis (SSc) and compare them to those without SSc, using data from the National Inpatient Sample from years 2016 to 2019. Methods and results International Classification of Diseases, Tenth Revision diagnosis codes were used to identify hospitalized patients with a primary diagnosis of HF and secondary diagnoses of SSc from the National Inpatient Sample database from 2016 to 2019. Patients were divided into two groups: those with and without a secondary diagnosis of SSc. Baseline characteristics including demographics and co‐morbidities, outcomes of mortality, length of stay (LOS), and costs were compared between the two groups. Multivariable logistic regression analysis was performed to adjust for confounders and assess the impact of SSc on in‐hospital mortality, cost, and LOS. A total of 4 709 724 hospitalizations for HF were identified, with 8150 (0.17%) having a secondary diagnosis of SSc. These patients were predominantly female (82.3% vs. 47.8%; P = 0.01), younger (mean age of 67.4 vs. 71.4; P

Background and aims – We tested the integrity of Rhynchospora sect. Pleurostachys, understand its evolution and origins, and test the phylogenetic separation of three morphologically distinct taxa. Material and methods – We used the chloroplast marker trnL-F and the two nuclear markers nrETS and nrITS in our phylogenetic analysis. Key results – We demonstrated that R. sect. Pleurostachys is monophyletic and that the section is divided into two large clades with the early divergent species in each clade occurring outside the Atlantic Forest of Brazil. The molecular analysis also confirmed the distinctness of the three species new to science. Conclusions – Rhynchospora sect. Pleurostachys probably originated outside the Atlantic Forest but diversified there. The three taxa are described as species new to science: Rhynchospora barbosae, R. hamadryadis, and R. eremica, from the tropical semi-deciduous and deciduous forests of Bahia and Espírito Santo, Brazil, are illustrated and compared to other related species of the section.

Abstract The outcome of CRISPR-Cas-mediated genome modifications is dependent on DNA double-strand break (DSB) processing and repair pathway choice. Homology-directed repair (HDR) of protein-blocked DSBs requires DNA end resection that is initiated by the endonuclease activity of the MRE11 complex. Using reconstituted reactions, we show that Cas9 breaks are unexpectedly not directly resectable by the MRE11 complex. In contrast, breaks catalyzed by Cas12a are readily processed. Cas9, unlike Cas12a, bridges the broken ends, preventing DSB detection and processing by MRE11. We demonstrate that Cas9 must be dislocated after DNA cleavage to allow DNA end resection and repair. Using single molecule and bulk biochemical assays, we next find that the HLTF translocase directly removes Cas9 from broken ends, which allows DSB processing by DNA end resection or non-homologous end-joining machineries. Mechanistically, the activity of HLTF requires its HIRAN domain and the release of the 3′-end generated by the cleavage of the non-target DNA strand by the Cas9 RuvC domain. Consequently, HLTF removes the H840A but not the D10A Cas9 nickase. The removal of Cas9 H840A by HLTF explains the different cellular impact of the two Cas9 nickase variants in human cells, with potential implications for gene editing.

Introdução: A implementação de um programa hospitalar de Gestão do Sangue do Paciente (Patient Blood Management - PBM) envolve a “abordagem centrada no paciente, sistemática e baseada em evidências para melhorar os resultados do paciente, gerenciando e preservando o sangue do próprio paciente, ao mesmo tempo em que promove a segurança e o empoderamento do paciente.” O PBM não se caracteriza apenas como um protocolo isolado, requer a participação multidisciplinar. Objetivos: Descrever o processo de implementação do programa de PBM num hospital privado, com foco em pacientes submetidos a cirurgias cardíacas de grande porte, comparando os dados da fase inicial de implementação, quando as ações de melhoria que ainda estavam sendo desenvolvidas, com os dados da fase de consolidação, quando elas já estavam implementadas. Material e métodos: Foram implementadas diversas ações, incluindo o desenvolvimento de um protocolo compreendendo as fases de pré, intra e pós-operatório, incorporando os 3 pilares do PBM. Além disso, foram estruturadas intervenções educativas e de monitoramento de indicadores, com feedback para as equipes clínicas. Resultados: Na fase inicial da implementação, de setembro de 2022 a junho de 2023, foram monitorados 229 pacientes submetidos a cirurgia cardíaca. Na fase de consolidação, de julho de 2023 a abril de 2024, mais 294 pacientes foram avaliados. Na análise de práticas intraoperatórias de PBM, observou-se que o uso de antifibrinolíticos oscilou de 81% para 83% (aumento de 2,5%; p = 0,561). A utilização de autotransfusão intraoperatória, aumentou de 55% para 69% (incremento de 25,5%; p = 0,001). A adoção de tromboelastometria para guiar a hemostasia subiu de 40% para 57% (aumento de 42,5%; p < 0,001). Quanto ao uso de hemocomponentes, o número médio de concentrados de hemácia por paciente diminuiu de 1,46 para 1,31 (redução de 10,3%; p = 0,784). De forma similar, a proporção de pacientes transfundidos caiu de 44% para 39% (redução de 11,3%; p = 0,327). Na análise dos desfechos, a ocorrência de complicações pós-operatórias reduziu de 52% para 20% (redução de 60,8%; p < 0,001), enquanto a incidência de infecção pós-operatória diminuiu de 27% para 17% (redução de 37%; p = 0,005). O tempo médio de permanência na unidade de terapia intensiva reduziu-se de 4,7 dias para 4,3 (redução de 8,7%; p = 0,543). A mortalidade intra-hospitalar caiu de 4,8% para 4,1% (redução de 15%; p = 0,830). Discussão: Um programa de PBM foi implementado com foco nas cirurgias cardíacas, sendo que algumas equipes já utilizavam técnicas de PBM mesmo antes da fase inicial da implementação. A comparação do período de implementação com o de consolidação revelou melhora importante nas práticas intraoperatórias, traduzidas pelo aumento no uso de autotransfusão e tromboelastometria. Redução importante na taxa de complicações e de infecção pós-operatória foi observada. Como todo processo de mudança de cultura e de processos, verificou-se a necessidade de intensificar os esforços de educação continuada e padronização de procedimentos para obtenção de melhores desfechos clínicos. Conclusão: A implementação bem-sucedida de um programa de PBM é factível com a elaboração de protocolos, ações educativas e monitoramento de indicadores. Com abordagem assertiva, tais estratégias do PBM podem levar a melhores resultados clínicos, maior segurança dos pacientes e a utilização mais racional dos recursos sanguíneos.

In this study, the mechanical behavior of concrete structures reinforced with one and two layers of the Brazilian glass textile “AR-360” was verified through direct tensile tests. The analysis of the mechanical behavior of the structures was performed with stress-strain curves, evaluating the transition points of the formed stages, rupture mode, rupture stress and strain, as well as an analysis comparing experimental results with analytics. Regarding the results, there was a structural inability of the pieces reinforced with a layer of glass textile. In the samples in which reinforcement with two layers of the aforementioned material was evaluated, the three stages present in the stress-strain curves were correctly identified, as predicted in the literature for the structures evaluated.

Abstract:  Currently, around 50% (~178 million) of all Americans are missing at least 1 tooth, with 12% (~40 million) of Americans missing all of their teeth. Dental implants are a common solution to this issue; titanium is the leading material of choice. Zirconia ceramic dental implants are a suitable replacement for titanium, but are often criticized for their brittleness. By developing a tantalum functional gradient coating, we hope to increase the viability of zirconia ceramic materials for dental implants. Two sets of material were chosen to test the proprietary BruxZir tantalum ion solution: yttria stabilized zirconia (3Y-TZP) and alumina toughened zirconia (ATZ). Solutions with varying concentrations of 0%, 10%, 15%, and 30% weight percent tantalum were prepared, and the ceramic samples were coated and sintered. Each sample was analyzed through various characterization methods, including SEM, EDS, XRD, nanoindentation, and Raman spectroscopy to observe how the solution impacted each sample’s microstructure and mechanical properties. Although our results are still in progress, we found evidence of an inverse relationship between tantalum concentration and grain size.

Grassy ecosystems occupy 27% of Brazilian territory and have historically been neglected in conservation efforts and studies about biodiversity. The Campos Sulinos region in Southern Brazil includes open formations in the Pampa and Atlantic Forest domains. This region has the smallest coverage by protected areas in the country and presents high levels of biodiversity and endemism. Besides, this region is under threat due to croplands and exotic tree plantations. Here, we compile a plant population genetics and phylogeography dataset from published studies focusing on species native to the Campos Sulinos to synthesize findings in plant evolution and genetics to better understand the dynamics of genetic diversity in this open ecosystem. We found 58 works on 51 plant species, published from 2005 to 2022. Most studies used only one kind of molecular marker and few loci. The climatic changes during the Pleistocene are likely the main speciation driver either due to the dynamics between grassland and forest or marine transgressions. Overall, high genetic variability and clear structuring of populations were found for the species studied. Regions with high genetic diversity do not coincide with protected areas. We reinforce the necessity of considering high intraspecific genetic diversity and population structure into both conservation and restoration planning, as well as in research within the Campos Sulinos.

Summary: Timed feeding drives adipose browning, although the integrative mechanisms for the same remain unclear. Here, we show that twice-a-night (TAN) feeding generates biphasic oscillations of circulating insulin and leptin, representing their entrainment by timed feeding. Insulin and leptin surges lead to marked cellular, functional, and metabolic remodeling of subcutaneous white adipose tissue (sWAT), resulting in increased energy expenditure. Single-cell RNA-sequencing (scRNA-seq) analyses and flow cytometry demonstrate a role for insulin and leptin surges in innate lymphoid type 2 (ILC2) cell recruitment and sWAT browning, since sWAT depot denervation or loss of leptin or insulin receptor signaling or ILC2 recruitment each dampens TAN feeding-induced sWAT remodeling and energy expenditure. Consistently, recreating insulin and leptin oscillations via once-a-day timed co-injections is sufficient to favorably remodel innervated sWAT. Innervation is necessary for sWAT remodeling, since denervation of sWAT, but not brown adipose tissue (BAT), blocks TAN-induced sWAT remodeling and resolution of inflammation. In sum, reorganization of nutrient-sensitive pathways remodels sWAT and drives the metabolic benefits of timed feeding.

OBJECTIVE: Classification criteria for calcium pyrophosphate deposition (CPPD) disease will facilitate clinical research on this common crystalline arthritis. Our objective was to report on the first 2 phases of a 4-phase process for developing CPPD classification criteria. METHODS: CPPD classification criteria development is overseen by a 12-member steering committee. Item generation (phase I) included a scoping literature review of 5 literature databases and contributions from a 35-member combined expert committee and 2 patient research partners. Item reduction and refinement (phase II) involved a combined expert committee meeting, discussions among clinical, imaging, and laboratory advisory groups, and an item-rating exercise to assess the influence of individual items toward classification. The steering committee reviewed the modal rating score for each item (range -3 [strongly pushes away from CPPD] to +3 [strongly pushes toward CPPD]) to determine items to retain for future phases of criteria development. RESULTS: Item generation yielded 420 items (312 from the literature, 108 from experts/patients). The advisory groups eliminated items that they agreed were unlikely to distinguish between CPPD and other forms of arthritis, yielding 127 items for the item-rating exercise. Fifty-six items, most of which had a modal rating of +/- 2 or 3, were retained for future phases. As numerous imaging items were rated +3, the steering committee recommended focusing on imaging of the knee and wrist and 1 additional affected joint for calcification suggestive of CPP crystal deposition. CONCLUSION: A data- and expert-driven process is underway to develop CPPD classification criteria. Candidate items comprise clinical, imaging, and laboratory features.

Breast cancer brain metastasis (BCBM) typically results in an end-stage diagnosis and is hindered by a lack of brain-penetrant drugs. Tumors in the brain rely on the conversion of acetate to acetyl-CoA by the enzyme acetyl-CoA synthetase 2 (ACSS2), a key regulator of fatty acid synthesis and protein acetylation. Here, we used a computational pipeline to identify novel brain-penetrant ACSS2 inhibitors combining pharmacophore-based shape screen methodology with absorption, distribution, metabolism, and excretion (ADME) property predictions. We identified compounds AD-5584 and AD-8007 that were validated for specific binding affinity to ACSS2. Treatment of BCBM cells with AD-5584 and AD-8007 leads to a significant reduction in colony formation, lipid storage, acetyl-CoA levels and cell survival in vitro. In an ex vivo brain-tumor slice model, treatment with AD-8007 and AD-5584 reduced pre-formed tumors and synergized with irradiation in blocking BCBM tumor growth. Treatment with AD-8007 reduced tumor burden and extended survival in vivo. This study identifies selective brain-penetrant ACSS2 inhibitors with efficacy towards breast cancer brain metastasis.

Geochemical negative emissions technologies (NETs) comprise a set of approaches to climate change mitigation that make use of alkaline minerals to remove and/or permanently store carbon dioxide (CO2) as solid carbonate minerals or dissolved ocean bicarbonate ions. This roadmap accompanies the comprehensive review of geochemical NETs by the same authors and offers guidance for the development and deployment of geochemical NETs at gigaton per year (Gt yr.−1) scale. We lay out needs and high-priority initiatives across six key elements required for the responsible and effective deployment of geochemical NETs: (i) technical readiness, (ii) social license, (iii) demand, (iv) supply chains, (v) human capital, and (vi) infrastructure. We put forward proposals for: specific initiatives to be undertaken; their approximate costs and timelines; and the roles that various actors could play in undertaking them. Our intent is to progress toward a working consensus among researchers, practitioners, and key players about initiatives that merit resourcing and action, primarily focusing on the near-term.