“Responsabilidade pré-contratual: os deveres de conduta na fase das negociações preliminares”

As negociações são uma fase na qual se mantém uma relação especial, em que, apesar de ainda não haver um contrato efetivamente concluído entre os envolvidos, estes travam um intenso contato, havendo grande troca de informações e maior possibilidade de interferência na esfera jurídica alheia. A especialidade que marca esse período requer uma proteção maior do que a conferida pelo princípio do “neminem laedere”, surgindo, assim, a responsabilidade civil pré-contratual. O caso mais famoso e mais frequente de responsabilidade civil pré-contratual é o que envolve um abandono abrupto de negociações quando já se havia criado no outro negociante, a legítima expectativa de contratação. No entanto, a doutrina aponta que a responsabilidade pré-contratual é mais ampla, abarcando todas as situações nas quais deveres anexos oriundos da boa-fé objetiva são descumpridos na fase pré-contratual. O presente trabalho busca, assim, realizar pesquisa jurisprudencial para verificar se há casos concretos que apontem para responsabilidade pré-contratual sem que seja causada pelo abrupto abandono de tratativas

Acute conversion of patient-derived Duchenne muscular dystrophy iPSC into myotubes reveals constitutive and inducible over-activation of TGFβ-dependent pro-fibrotic signaling

Background In Duchenne muscular dystrophy (DMD), DYSTROPHIN deficiency exposes myofibers to repeated cycles of contraction/degeneration, ultimately leading to muscle loss and replacement by fibrotic tissue. DMD pathology is typically exacerbated by excessive secretion of TGFβ and consequent accumulation of pro-fibrotic components of the extra-cellular matrix (ECM), which in turn impairs compensatory regeneration and complicates the efficacy of therapeutic strategies. It is currently unclear whether DMD skeletal muscle fibers directly contribute to excessive activation of TGFβ. Development of skeletal myofibers from DMD patient-derived induced pluripotent stem cells (iPSC), as an “in dish” model of disease, can be exploited to determine the myofiber contribution to pathogenic TGFβ signaling in DMD and might provide a screening platform for the identification of anti-fibrotic interventions in DMD. Methods We describe a rapid and efficient method for the generation of contractile human skeletal muscle cells from DMD patient-derived hiPSC, based on the inducible expression of MyoD and BAF60C (encoded by SMARCD3 gene), using an enhanced version of piggyBac (epB) transposone vectors. DMD iPSC-derived myotubes were tested as an “in dish” disease model and exposed to environmental and mechanical cues that recapitulate salient pathological features of DMD. Results We show that DMD iPSC-derived myotubes exhibit a constitutive activation of TGFβ-SMAD2/3 signaling. High-content screening (HCS)-based quantification of nuclear phosphorylated SMAD2/3 signal revealed that DMD iPSC-derived myotubes also exhibit increased activation of the TGFβ-SMAD2/3 signaling following exposure to either recombinant TGFβ or electrical pacing-induced contraction. Conclusions Acute conversion of DMD patient-derived iPSC into skeletal muscles, by the ectopic expression of MyoD and BAF60C, provides a rapid and reliable protocol for an “in dish” DMD model that recapitulates key pathogenic features of disease pathology, such as the constitutive activation of the TGFβ/SMAD signaling as well as the deregulated response to pathogenic stimuli, e.g., ECM-derived signals or mechanical cues. Thus, this model is suitable for the identification of new therapeutic targets in DMD patient-specific muscles

Histological Characteristics, Fatty Acid Composition of Lipid Fractions, and Cholesterol Content of Semimembranosus and Triceps Brachii Muscles in Maremmana and Limousine Bovine Breeds

This study examined the histological properties of Semimembranosus and Triceps brachii muscle in two different bovine breeds, Maremmana (an autochthonous breed from Tuscany, Italy) and Limousine. The animals were grazed in two adjoining pastures, received the same feed supplementation and were weighed monthly. The experimental period lasted from weaning (six months old) to slaughter (19 months old). Muscle samples were collected immediately after slaughter, before carcass cooling. Regarding the histological properties, the number of fibres (TNF), mean sarcolemma perimeter (MSP), cross section area (CSA), and total sarcolemma perimeter (TSP) were determined. Samples were also analysed for proximate composition, fatty acid profile of total lipids, phospholipids and neutral lipids and for total cholesterol content. Breed was a significant variation factor for the performance parameter and histological muscle fibre properties. Interestingly, despite that Maremmana being a less extensively genetically improved breed than Limousine, it showed higher weight at slaughter (+18%) and daily weight gain (+19%). Maremmana also showed smaller muscle fibres than Limousine and, consequently, the TSP was higher. This difference affected the lipid fraction distribution (Limousine was higher in phospholipids and lower in neutral lipids than Maremmana) and, consequently, the fatty acid composition of total lipids (Limousine was high in polyunsaturated fatty acids, while Maremmana was high in monounsaturated fatty acids). The results of this experiment highlight the importance of environmental and management conditions on the full expression of genotypic potentia

Next-generation sequencing approach to hyperCKemia: A 2-year cohort study

Next-generation sequencing (NGS) was applied in molecularly undiagnosed asymptomatic or paucisymptomatic hyperCKemia to investigate whether this technique might allow detection of the genetic basis of the condition

Carbon dioxide fluxes in Alpine grasslands at the Nivolet Plain, Gran Paradiso National Park, Italy 2017–2023

The version of record of this article, first published in [Scientific Data], is available online at Publisher’s website: http://dx.doi.org/10.1038/s41597-024-03374-1We introduce a georeferenced dataset of Net Ecosystem Exchange (NEE), Ecosystem Respiration (ER) and meteo-climatic variables (air and soil temperature, air relative humidity, soil volumetric water content, pressure, and solar irradiance) collected at the Nivolet Plain in Gran Paradiso National Park (GPNP), western Italian Alps, from 2017 to 2023. NEE and ER are derived by measuring the temporal variation of CO2 concentration obtained by the enclosed chamber method. We used a customised portable non-steady-state dynamic flux chamber, paired with an InfraRed Gas Analyser (IRGA) and a portable weather station, measuring CO2 fluxes at a number of points (around 20 per site and per day) within five different sites during the snow-free season (June to October). Sites are located within the same hydrological basin and have different geological substrates: carbonate rocks (site CARB), gneiss (GNE), glacial deposits (GLA, EC), alluvial sediments (AL). This dataset provides relevant and often missing information on high-altitude mountain ecosystems and enables new comparisons with other similar sites, modelling developments and validation of remote sensing data.This work was funded by the H2020 projects ECOPOTENTIAL (grant number: 641762), e-shape (grant number: 820852), eLTER PLUS (grant number: 871128), by the Italian National Biodiversity Future Center (NBFC), National Recovery and Resilience Plan (NRRP; mission 4, component 2, investment 1.4 of the Ministry of University and Research, funded by the European Union–NextGenerationEU; project code CN00000033), and by the ITINERIS NRRP Italian infrastructure project (project code No. IR0000032 - ESFRI Environment)

Measurement of χ c1 and χ c2 production with s√ = 7 TeV pp collisions at ATLAS

The prompt and non-prompt production cross-sections for the χ c1 and χ c2 charmonium states are measured in pp collisions at s√ = 7 TeV with the ATLAS detector at the LHC using 4.5 fb−1 of integrated luminosity. The χ c states are reconstructed through the radiative decay χ c → J/ψγ (with J/ψ → μ + μ −) where photons are reconstructed from γ → e + e − conversions. The production rate of the χ c2 state relative to the χ c1 state is measured for prompt and non-prompt χ c as a function of J/ψ transverse momentum. The prompt χ c cross-sections are combined with existing measurements of prompt J/ψ production to derive the fraction of prompt J/ψ produced in feed-down from χ c decays. The fractions of χ c1 and χ c2 produced in b-hadron decays are also measured

Congenital myopathies: Clinical phenotypes and new diagnostic tools

Congenital myopathies are a group of genetic muscle disorders characterized clinically by hypotonia and weakness, usually from birth, and a static or slowly progressive clinical course. Historically, congenital myopathies have been classified on the basis of major morphological features seen on muscle biopsy. However, different genes have now been identified as associated with the various phenotypic and histological expressions of these disorders, and in recent years, because of their unexpectedly wide genetic and clinical heterogeneity, next-generation sequencing has increasingly been used for their diagnosis. We reviewed clinical and genetic forms of congenital myopathy and defined possible strategies to improve cost-effectiveness in histological and imaging diagnosis

HemaMax™, a Recombinant Human Interleukin-12, Is a Potent Mitigator of Acute Radiation Injury in Mice and Non-Human Primates

HemaMax, a recombinant human interleukin-12 (IL-12), is under development to address an unmet medical need for effective treatments against acute radiation syndrome due to radiological terrorism or accident when administered at least 24 hours after radiation exposure. This study investigated pharmacokinetics, pharmacodynamics, and efficacy of m-HemaMax (recombinant murine IL-12), and HemaMax to increase survival after total body irradiation (TBI) in mice and rhesus monkeys, respectively, with no supportive care. In mice, m-HemaMax at an optimal 20 ng/mouse dose significantly increased percent survival and survival time when administered 24 hours after TBI between 8–9 Gy (p<0.05 Pearson's chi-square test). This survival benefit was accompanied by increases in plasma interferon-γ (IFN-γ) and erythropoietin levels, recovery of femoral bone hematopoiesis characterized with the presence of IL-12 receptor β2 subunit–expressing myeloid progenitors, megakaryocytes, and osteoblasts. Mitigation of jejunal radiation damage was also examined. At allometrically equivalent doses, HemaMax showed similar pharmacokinetics in rhesus monkeys compared to m-HemaMax in mice, but more robustly increased plasma IFN-γ levels. HemaMax also increased plasma erythropoietin, IL-15, IL-18, and neopterin levels. At non-human primate doses pharmacologically equivalent to murine doses, HemaMax (100 ng/Kg and 250 ng/Kg) administered at 24 hours after TBI (6.7 Gy/LD50/30) significantly increased percent survival of HemaMax groups compared to vehicle (p<0.05 Pearson's chi-square test). This survival benefit was accompanied by a significantly higher leukocyte (neutrophils and lymphocytes), thrombocyte, and reticulocyte counts during nadir (days 12–14) and significantly less weight loss at day 12 compared to vehicle. These findings indicate successful interspecies dose conversion and provide proof of concept that HemaMax increases survival in irradiated rhesus monkeys by promoting hematopoiesis and recovery of immune functions and possibly gastrointestinal functions, likely through a network of interactions involving dendritic cells, osteoblasts, and soluble factors such as IL-12, IFN-γ, and cytoprotectant erythropoietin

An explainable model of host genetic interactions linked to COVID-19 severity

We employed a multifaceted computational strategy to identify the genetic factors contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing (WES) dataset of a cohort of 2000 Italian patients. We coupled a stratified k-fold screening, to rank variants more associated with severity, with the training of multiple supervised classifiers, to predict severity based on screened features. Feature importance analysis from tree-based models allowed us to identify 16 variants with the highest support which, together with age and gender covariates, were found to be most predictive of COVID-19 severity. When tested on a follow-up cohort, our ensemble of models predicted severity with high accuracy (ACC = 81.88%; AUCROC = 96%; MCC = 61.55%). Our model recapitulated a vast literature of emerging molecular mechanisms and genetic factors linked to COVID-19 response and extends previous landmark Genome-Wide Association Studies (GWAS). It revealed a network of interplaying genetic signatures converging on established immune system and inflammatory processes linked to viral infection response. It also identified additional processes cross-talking with immune pathways, such as GPCR signaling, which might offer additional opportunities for therapeutic intervention and patient stratification. Publicly available PheWAS datasets revealed that several variants were significantly associated with phenotypic traits such as "Respiratory or thoracic disease", supporting their link with COVID-19 severity outcome.A multifaceted computational strategy identifies 16 genetic variants contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing dataset of a cohort of Italian patients