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ATTR amyloidosis is caused by the deposition of transthyretin in the form of amyloid fibrils in virtually every organ of the body, including the heart. This systemic deposition leads to a phenotypic variability that has not been molecularly explained yet. In brain amyloid conditions, previous studies suggest an association between clinical phenotype and the molecular structures of their amyloid fibrils. Here we investigate whether there is such an association in ATTRv amyloidosis patients carrying the mutation I84S. Using cryo-electron microscopy, we determined the structures of cardiac fibrils extracted from three ATTR amyloidosis patients carrying the ATTRv-I84S mutation, associated with a consistent clinical phenotype. We found that in each ATTRv-I84S patient, the cardiac fibrils exhibited different local conformations, and these variations can co-exist within the same fibril. Our finding suggests that one amyloid disease may associate with multiple fibril structures in systemic amyloidoses, calling for further studies.

Abstract Low injectivity is often experienced in geothermal doublets installed in sandstone reservoirs. This even led to a shutdown of the Mezőberény (Hungary) geothermal site. An on-site campaign was carried out in January 2021 to prepare a stimulation aiming to enhance the transmissivity of the sedimentary reservoir and the near-wellbore zone of this site. Previous studies have concluded that insufficient injectivity may be linked to a high skin effect in the near well-bore zone and pore clogging in combination with the low net sandstone content of the fluvio-deltaic reservoir. A chemical soft stimulation based on the injection of hydrochloric acid (HCl) was successfully used to unclog and recover the well injectivity. Despite such empirical evidence, the geochemical mechanisms leading to both, detrimental formation of clogging and the HCl-driven transmissivity restoration, have not yet been elucidated. This work presents the results of a novel analysis aiming at (a) predicting the dominant type of clogging forming in the near-well bore zone; (b) quantifying the drop in hydraulic conductivity as clogging occurs; and (c) supporting the optimization of the HCl dosage during the chemical soft stimulation. The study is supported by new experimental datasets never presented before from the Mezőberény site and a geochemical model set-up simulating the main mechanisms involved in the clogging and unclogging processes. It is concluded that the biofilm formation was the dominant, while the precipitation of calcite and amorphous ferrihydrite—later reduced to magnetite by microbes—was the secondary clogging mechanism: In the long-term (yearly scale) simulating the hydraulic conductivity showed a decline with forming scales; therefore, biofilm was presumably responsible for the experienced rapid (1 month) clogging. When modelling the chemical stimulation, the estimated amount of precipitated minerals was dissolved already with 2.5 mol of HCl per liter of water (~ 10 m/m%). Therefore, the 20 m/m% of HCl chosen during the field campaign might had a beneficial effect dissolving the potentially higher amount of scaling and/or the carbonate minerals of the matrix near the wellbore. Overall, it is concluded that the chemical and the microbial analyses together with the geochemical model were critical to tailor the remediation attempts and to propose further development or reconstruction of the surface system before going into operation to prevent recurrent impairments. Our findings highlight the importance of interactions of various clogging mechanisms with each other as well as with the reservoir processes and provide approaches to tackle the issue of injectivity drop by characterizing and quantifying their effects.

Abstract Community-acquired pneumonia (CAP) is an acute infection of the lung parenchyma acquired outside the hospital or other healthcare settings, typically affecting previously healthy individuals. This intersociety consensus aims to provide evidence-based recommendations for the antibiotic treatment of mild to moderate CAP in previously healthy children in Italy. A systematic review was conducted to identify the most recent and relevant evidence. Embase, Scopus, PubMed, and Cochrane databases were systematically screened, with a date restriction from 2012 to April 2024, but without language limitations. The review included studies conducted in high-income countries on antibiotic therapy in children over 3 months of age diagnosed with mild-moderate CAP. The certainty of evidence was assessed using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methods. The final recommendations were obtained through a Delphi consensus of an expert panel. Amoxicillin is the first-line treatment if the child is at least immunized against Haemophilus influenzae type b (low/very low quality of evidence, strong recommendations), while amoxicillin-clavulanate or second- or third-generation cephalosporins should be prescribed for those unimmunized or with incomplete immunization coverage for both H. influenzae type b and Streptococcus pneumoniae (low/very low quality of evidence, strong recommendations). Macrolides should be considered in addition to amoxicillin in children over 5 years old, if symptoms persist and the clinical condition remains good after 48 h of therapy (low/very low quality of evidence, strong recommendations). The dosage of amoxicillin is 90 mg/kg/day divided in three doses, although two doses could be considered to improve compliance (moderate quality of evidence, weak recommendations). A five-day duration of therapy is recommended, with clinical monitoring and re-assessment approximately 72 h after the start of antibiotic treatment to evaluate symptom resolution (moderate quality of evidence, strong recommendations). To improve the management of CAP in pediatric patients, we have developed this consensus based on a thorough review of the best available evidence and extensive discussions with an expert panel. However, further efforts are needed. Future research should focus on enhancing diagnostic accuracy, optimizing antibiotic utilization, comparing the efficacy of different antibiotic regimens, and determining the optimal dosage and duration of treatment in different setting.

Abstract Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) is a rare but life-threatening cutaneous drug reaction mediated by human leukocyte antigen (HLA) class I-restricted CD8+ T cells. For unbiased assessment of cellular immunopathogenesis, here we perform single-cell (sc) transcriptome, surface proteome, and T cell receptor (TCR) sequencing on unaffected skin, affected skin, and blister fluid from 15 SJS/TEN patients. From 109,888 cells, we identify 15 scRNA-defined subsets. Keratinocytes express markers indicating HLA class I-restricted antigen presentation and appear to trigger the proliferation of and killing by cytotoxic CD8+ tissue-resident T cells that express granulysin, granzyme B, perforin, LAG3, CD27, and LINC01871, and signal through the PKM, MIF, TGFβ, and JAK-STAT pathways. In affected tissue, cytotoxic CD8+ T cells express private expanded and unexpanded TCRαβ that are absent or unexpanded in unaffected skin, and mixed populations of macrophages and fibroblasts express pro-inflammatory markers or those favoring repair. This data identifies putative cytotoxic TCRs and therapeutic targets.

Aim – to determine additional risks of developing arterial thrombotic and thromboembolic complications in bradysystolic AF and substantiate the results using modeling of intra-arterial hemodynamics. Material and methods. A single-center prospective study involving 252 patients: 146 in the main group, 106 in the control group. The main group was divided into 2 subgroups: 1A subgroup RR ECG interval 1.5 seconds; 2B subgroup RR≥1.5 seconds. A comprehensive examination of the patients was carried out. The second stage is prospective comprising an analysis of the development of arterial thrombotic and thromboembolic complications over 1 year. Experimental modeling was carried out using the “Device for simulating intra-arterial circulation”. Results. Thrombotic and thromboembolic complications were more common in subgroup 1B (OR=8.287 (2.287; 30.040); z=3.219; p=0.001). When analyzing the main parameters of the hemodynamics of the main arteries, the first pulse wave, coming after a long pause of 1.5 seconds or more in AF, was accompanied by a statistically significant increase in all of analyzed parameters. In the experiment, when simulating AF, the intensity of the mechanical impact of the free end of the thread on the wall of the rotameter was maximum when the pause between pulse waves was 1.5 seconds or more (9.70 ± 2.52 mm). At this moment, the piezocrystalline pressure sensor recorded the maximum increase in pressure inside the rotameter tube by an average of 56%. Conclusions. Bradysystole in AF is associated with a significantly higher likelihood of developing long-term thromboembolic events. The first pulse wave, coming after a long pause between ventricular contractions during AF, leads to a significant increase in the main parameters of the hemodynamics of the main arteries (linear velocity of blood flow, volumetric blood flow). When monitoring heart rate in AF, it is necessary to avoid bradysystole with pauses between ventricular contractions of 1.5 seconds or more, due to a higher risk of stroke, myocardial infarction, and distal arterial embolism in other vascular regions.

Abstract Specialized function gradient computing hardware could greatly improve the performance of state-of-the-art optimization algorithms. Prior work on such hardware, performed in the context of Ising Machines and related concepts, is limited to quadratic polynomials and not scalable to commonly used higher-order functions. Here, we propose an approach for massively parallel gradient calculations of high-degree polynomials, which is conducive to efficient mixed-signal in-memory computing circuit implementations and whose area scales proportionally with the product of the number of variables and terms in the function and, most importantly, independent of its degree. Two flavors of such an approach are proposed. The first is limited to binary-variable polynomials typical in combinatorial optimization problems, while the second type is broader at the cost of a more complex periphery. To validate the former approach, we experimentally demonstrated solving a small-scale third-order Boolean satisfiability problem based on integrated metal-oxide memristor crossbar circuits, with competitive heuristics algorithm. Simulation results for larger-scale, more practical problems show orders of magnitude improvements in area, speed and energy efficiency compared to the state-of-the-art. We discuss how our work could enable even higher-performance systems after co-designing algorithms to exploit massively parallel gradient computation.

Abstract Background Considering that school environments are important and have the potential to promote an active lifestyle with direct impacts on physical, motor, cognitive, affective, and psychological aspects, the objective of this study was to evaluate the effect of a physical exercise program in the school context. Methods A parallel, two-arm experimental research project carried out over 19 weeks, with two non-consecutive weekly classes of 60 min, with the inclusion of PROFIT-BR in the initial 15 min of the class followed by 40 min based on specific physical education skills on indicators of physical fitness in children. Estimates of excess weight and visceral fat will be measured and assessed by body mass index and waist-to-height ratio, respectively, and cardiorespiratory fitness by performance in the six-minute run/walk test. Flexibility will be assessed by the sit and reach test; and muscle strength measured by the number of sit-ups in one minute. The 2 kg medicine ball throw and the horizontal jump will be used to measure upper and lower limb power, respectively, the square test used as a measure of agility, and the 20-meter run test used to measure speed. To analyze the differences between baseline and post-intervention values, repeated measures ANOVA adjusted for somatic maturation will be used. The magnitude of the effect size will be estimated by “partial eta squared”. Statistical analyses will be carried out using SPSS version 24.0 software and a 5% probability of error in the analyses will be accepted. Discussion The main theoretical assumption of the proposal is the cause-and-effect relationship between the regular practice of physical exercise in childhood and adolescence and several parameters of health and motor performance. The elaborated program has a lot of applicability because it was wondering from the perspective of the school, in terms of materials, space, and objectives of physical education.

Molecular docking is a widely used technique in drug discovery to predict the binding mode of a given ligand to its target. However, the identification of the near-native binding pose in docking experiments still represents a challenging task as the scoring functions currently employed by docking programs are parametrized to predict the binding affinity, and, therefore, they often fail to correctly identify the ligand native binding conformation. Selecting the correct binding mode is crucial to obtaining meaningful results and to conveniently optimizing new hit compounds. Deep learning (DL) algorithms have been an area of a growing interest in this sense for their capability to extract the relevant information directly from the protein-ligand structure. Our review aims to present the recent advances regarding the development of DL-based pose selection approaches, discussing limitations and possible future directions. Moreover, a comparison between the performances of some classical scoring functions and DL-based methods concerning their ability to select the correct binding mode is reported. In this regard, two novel DL-based pose selectors developed by us are presented.

Abstract In silico prediction of xenobiotic metabolism is an important strategy to accelerate the drug discovery process, as candidate compounds often fail in clinical phases due to their poor pharmacokinetic profiles. Here we present MetaQM, a dataset of quantum-mechanical (QM) optimized metabolic substrates, including force field parameters, electronic and physicochemical properties. MetaQM comprises 2054 metabolic substrates extracted from the MetaQSAR database. We provide QM-optimized geometries, General Amber Force Field (FF) parameters for all studied molecules, and an extended set of structural and physicochemical descriptors as calculated by DFT and PM7 methods. The generated data can be used in different types of analysis. FF parameters can be applied to perform classical molecular mechanics calculations as exemplified by the validating molecular dynamics simulations reported here. The calculated descriptors can represent input features for developing improved predictive models for metabolism and drug design, as exemplified in this work. Finally, the QM-optimized molecular structures are valuable starting points for both ligand- and structure-based analyses such as pharmacophore mapping and docking simulations.

Abstract Background Poly (ADP-ribose) polymerase 1 and 2 (PARP1/2) inhibitors (PARPi) are targeted therapies approved for homologous recombination repair (HRR)-deficient breast, ovarian, pancreatic, and prostate cancers. Since inhibition of PARP1 is sufficient to cause synthetic lethality in tumors with homologous recombination deficiency (HRD), PARP1 selective inhibitors such as saruparib (AZD5305) are being developed. It is expected that selective PARP1 inhibition leads to a safer profile that facilitates its combination with other DNA damage repair inhibitors. Here, we aimed to characterize the antitumor activity of AZD5305 in patient-derived preclinical models compared to the first-generation PARP1/2 inhibitor olaparib and to identify mechanisms of resistance. Methods Thirteen previously characterized patient-derived tumor xenograft (PDX) models from breast, ovarian, and pancreatic cancer patients harboring germline pathogenic alterations in BRCA1, BRCA2, or PALB2 were used to evaluate the efficacy of AZD5305 alone or in combination with carboplatin or an ataxia telangiectasia and Rad3 related (ATR) inhibitor (ceralasertib) and compared it to the first-generation PARPi olaparib. We performed DNA and RNA sequencing as well as protein-based assays to identify mechanisms of acquired resistance to either PARPi. Results AZD5305 showed superior antitumor activity than the first-generation PARPi in terms of preclinical complete response rate (75% vs. 37%). The median preclinical progression-free survival was significantly longer in the AZD5305-treated group compared to the olaparib-treated group (> 386 days vs. 90 days). Mechanistically, AZD5305 induced more replication stress and genomic instability than the PARP1/2 inhibitor olaparib in PARPi-sensitive tumors. All tumors at progression with either PARPi (39/39) showed increase of HRR functionality by RAD51 foci formation. The most prevalent resistance mechanisms identified were the acquisition of reversion mutations in BRCA1/BRCA2 and the accumulation of hypomorphic BRCA1. AZD5305 did not sensitize PDXs with acquired resistance to olaparib but elicited profound and durable responses when combined with carboplatin or ceralasertib in 3/6 and 5/5 models, respectively. Conclusions Collectively, these results show that the novel PARP1 selective inhibitor AZD5305 yields a potent antitumor response in PDX models with HRD and delays PARPi resistance alone or in combination with carboplatin or ceralasertib, which supports its use in the clinic as a new therapeutic option.

Abstract ATTR amyloidosis results from the conversion of transthyretin into amyloid fibrils that deposit in tissues causing organ failure and death. This conversion is facilitated by mutations in ATTRv amyloidosis, or aging in ATTRwt amyloidosis. ATTRv amyloidosis exhibits extreme phenotypic variability, whereas ATTRwt amyloidosis presentation is consistent and predictable. Previously, we found unique structural variabilities in cardiac amyloid fibrils from polyneuropathic ATTRv-I84S patients. In contrast, cardiac fibrils from five genotypically different patients with cardiomyopathy or mixed phenotypes are structurally homogeneous. To understand fibril structure’s impact on phenotype, it is necessary to study the fibrils from multiple patients sharing genotype and phenotype. Here we show the cryo-electron microscopy structures of fibrils extracted from four cardiomyopathic ATTRwt amyloidosis patients. Our study confirms that they share identical conformations with minimal structural variability, consistent with their homogenous clinical presentation. Our study contributes to the understanding of ATTR amyloidosis biopathology and calls for further studies.

Pediatric Autoimmune Neuropsychiatric Disorder Associated with Streptococcal Infections (PANDAS) and Pediatric Acute-onset Neuropsychiatric Syndrome (PANS) are broad diagnoses that encompass a range of sudden-onset neuropsychiatric symptoms in children, which can include obsessive-compulsive disorder (OCD), tics, anxiety, emotional instability, and cognitive difficulties. Unlike PANDAS, PANS is not strictly linked to group A streptococcal infections but can be triggered by various infectious or environmental factors. Lights and shadows remain upon the management of children with PANS and PANDAS and there is no clear consensus regarding definition, diagnostic criteria, treatment, and follow-up. The aim of the present study was to evaluate the level of agreement on PANS and PANDAS definition, diagnostic criteria, treatment and follow-up and to assess on the basis of recent studies whether there is a need to modify the current recommendations used by primary care pediatricians and hospital pediatricians in clinical practice in order to improve outcomes. Using the Delphi method, this consensus provides shared indications on PANS and PANDAS management in pediatric age, based on the most updated literature. This work represents, in our opinion, the most complete and up-to-date information on the diagnosis of PANS and PANDAS, as well as consensus statements about several aspects of clinical care. Undoubtedly, more randomized and controlled trials are needed in the pediatric population to better define the best management, also in terms of adequate follow-up examinations and period of observation.

Introduction: The neutrophil/lymphocyte ratio (NLR) and the platelet/lymphocyte ratio (PLR) are markers of inflammation and prognosis in systemic diseases. This study determined the association between hematological indices, and disease activity in patients with systemic lupus erythematosus (SLE). Methods: This prospective, observational and analytical cross-sectional study included 87 Paraguayan patients diagnosed with SLE, according to the ACR/EULAR 2019 criteria. We investigate four hematological indices: neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), eosinophil/lymphocyte ratio (ELR), and monocytes/lymphocyte ratio (MLR). Results: Eighty-seven percent were female, with a median age of 34.9 ±12.3 years. SLEDAI mean of 3.68 ± 4.76 was found. SLEDAI mean was 3.68 ± 4.76. Positivity of anti-DNAds was found in 52% of the patients. CRP mean was 3.5 ± 4.6. Regarding values of hematological indices, we found: NLR mean of 2.4 ± 1.6 was obtained, ELR 0.07 ± 0.09, MRL 0.07 ± 0.07 and PLR 155.89 ± 80.69. A significant positive correlation was found with SLEDAI and NLR (r=0.34, p=0.001). Based on the ROC curve, the best cut-off value for our patient cohort capable of predicting patients with high activity is for NLR values ? 2.283 (AUC: 0.66). Conclusion: NLR could be a valuable and accessible biomarker to identify elevated activity in patients with SLE.

Abstract The escalating drug resistance among microorganisms underscores the urgent need for innovative therapeutic strategies and a comprehensive understanding of bacteria's defense mechanisms against oxidative stress and antibiotics. Among the recently discovered barriers, the endogenous production of hydrogen sulfide (H2S) via the reverse transsulfuration pathway, emerges as a noteworthy factor. In this study, we have explored the catalytic capabilities and crystal structure of cystathionine γ-lyase from Pseudomonas aeruginosa (PaCGL), a multidrug-opportunistic pathogen chiefly responsible for nosocomial infections. In addition to a canonical l-cystathionine hydrolysis, PaCGL efficiently catalyzes the production of H2S using l-cysteine and/or l-homocysteine as alternative substrates. Comparative analysis with the human enzyme and counterparts from other pathogens revealed distinct structural features within the primary enzyme cavities. Specifically, a distinctly folded entrance loop could potentially modulate the access of substrates and/or inhibitors to the catalytic site. Our findings offer significant insights into the structural evolution of CGL enzymes across different pathogens and provide novel opportunities for developing specific inhibitors targeting PaCGL.

Abstract The aim of the current study was to investigate the influence of both intra- and interspecific audiences on dogs' facial expressions and behaviours. Forty-six dogs were exposed to three test conditions in which a food reward, initially available, was denied when in the presence of either a human (Human condition) or a dog audience (Dog condition), or in the absence of a visible audience (Non-social condition). Salivary cortisol was collected to evaluate the stress/arousal activation in the different conditions. Compared to the Non-social condition, the presence of a conspecific evoked more facial expressions, according to the DogFACS (Facial Action Coding System, an anatomically based tool to analyze facial expressions in domestic dogs), (EAD105—Ears downward), displacement behaviours (AD137—Nose licking, AD37—Lip wiping), tail wagging, whining, and panting (AD126). When facing a conspecific, dogs assumed a more avoidant attitude, keeping a distance and not looking at the stimuli, compared to when in the presence of the human partner. Dogs also exhibited more facial expressions (EAD102—Ears Adductor, EAD104—Ears Rotator), displacement behaviours (AD137—Nose licking, AD37—Lip wiping), panting (AD126) and whining when facing the conspecific than the human partner. Post-test cortisol was not influenced by any condition, and no association between pre-test cortisol and behavioural variables was found, thus strong differences in the levels of stress/arousal were unlikely to be responsible for differences in behavior between conditions. Considering the current results in the context of the available literature, we suggest that the higher displacement behaviors exhibited with the conspecifics were likely due to an increased level of uncertainty regarding the situations.

Subsidence refers to the gradual lowering or sudden sinking of the ground surface and is known to impact human lives in terms of damages to the infrastructures, utility lines, and buildings as well as changes in the surficial drainage systems and groundwater conditions. The impacts of land subsidence will be greater in the future, considering the sea level rise, population growth, intensification of coastal erosion and extreme events, as well as increase in flood risk or freshwater salinization, mostly in coastal cities. The main aim of this work is to provide an open-source, peer-reviewed, and comprehensive database identifying the main and secondary causes of land subsidence in 143 coastal cities. We highlight the potential impacts of subsidence that are still unknown in some at-risk cities and non-existence of mitigation measures. The database additionally shows that mitigation measures, specifically those addressing subsidence due to groundwater extraction, have proven successful in the past. The proposed database aims to increase the knowledge on the subsidence phenomenon and also global awareness of land subsidence issues among researchers, the scientific community, stakeholders, and policymakers in terms of urban planning and development.

O presente artigo apresenta um material didático que possa auxiliar professores de biologia a elaborarem aulas mais participativas para os alunos surdos, a fim de proporcionar uma aprendizagem atrativa e significativa. Para tal, foi realizada uma pesquisa exploratória com professores de Biologia e, em seguida, foi realizada uma pesquisa documental, na qual buscou-se selecionar na plataforma Google canais do Youtube, jogos e aplicativos que pudessem contribuir para as práticas docentes. Os materiais selecionados foram reunidos em um e-book, junto à uma proposta de Sequência Didática (SD), sendo posteriormente avaliado por professores de biologia. O estudo possibilitou a percepção de que os docentes carecem de materiais que possam auxiliar a aprendizagem dos alunos surdos, assim como a importância das tecnologias digitais visuais para esse processo, sendo o e-book elaborado considerado potencialmente capaz de contribuir para as práticas pedagógicas inclusivas.

Sea fennel is a minor crop of emerging socioeconomic importance in the Mediterranean region. Despite its potential, there has been no assessment of its environmental impacts to support its perceived sustainability for broader promotion. Using a life cycle assessment (LCA), we evaluated the environmental performance of sea fennel production in an open field. A cradle-to-farm gate assessment was conducted using a functional unit (FU) of 1 kg of fresh sea fennel at the farm gate. The system boundaries encompassed the nursery, cultivation, and waste management phases. Primary data was collected from farms in the Marche region of Italy. The environmental impacts were assessed using the Environmental Footprint (EF) 3.0 midpoint life cycle impact assessment method. From the results, the primary hotspot input was plastic mulch, with a relative contribution of over 50% across most impact categories. Increasing the recycling rate from 30% to 70% substantially improved the environmental performance of sea fennels, reiterating the need for further investment in plastic recycling. This study provides insights into the environmental sustainability of sea fennel production. Findings demonstrate that sea fennel cultivation offers a promising path toward sustainable biomass production, underscoring the prospects for its commercial exploitation.