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Some proteins perform their biological functions by changing their material states through liquid-liquid phase separation. Upon phase separation, the protein condenses into a concentrated liquid phase and sometimes into a gel phase, changing its dynamic properties and intermolecular interactions, thereby regulating cellular functions. Although the biological significance of this phenomenon has been widely recognized by researchers, there is still a lack of a comprehensive understanding of the structural and dynamic properties of the protein in the condensed phase. In this phase, molecules usually contain domains with varied dynamic properties and undergo intermediate exchanges. Magic angle spinning (MAS) solid-state NMR (SSNMR) experiments are very powerful in studying rigid protein polymers such as amyloid. The incorporation of solution-like experiments into SSNMR and the development of J-coupling based MAS SSNMR techniques extend its ability to study partially mobile segments of proteins in a condensed liquid or gel phase which are not visible by solution NMR or dipolar-coupling based SSNMR. Therefore, it has been applied in studying protein condensation and has provided very important information that is hard to obtain by other techniques.

Abstract Background Glioblastoma (GBM) is the most malignant among gliomas with an inevitable lethal outcome. The elucidation of the physiology and regulation of this tumor is mandatory to unravel novel target and effective therapeutics. Emerging concepts show that the minor subset of glioblastoma stem cells (GSCs) accounts for tumorigenicity, representing the true target for innovative therapies in GBM. Methods Here, we isolated and established functionally stable and steadily expanding GSCs lines from a large cohort of GBM patients. The molecular, functional and antigenic landscape of GBM tissues and their derivative GSCs was highlited in a side-by-side comprehensive genomic and transcriptomic characterization by ANOVA and Fisher’s exact tests. GSCs’ physio-pathological hallmarks were delineated by comparing over time in vitro and in vivo their expansion, self-renewal and tumorigenic ability with hierarchical linear models for repeated measurements and Kaplan–Meier method. Candidate biomarkers performance in discriminating GBM patients’ classification emerged by classification tree and patients’ survival analysis. Results Here, distinct biomarker signatures together with aberrant functional programs were shown to stratify GBM patients as well as their sibling GSCs population into TCGA clusters. Of importance, GSCs cells were demonstrated to fully resemble over time the molecular features of their patient of origin. Furthermore, we pointed out the existence of distinct GSCs subsets within GBM classification, inherently endowed with different self-renewal and tumorigenic potential. Particularly, classical GSCs were identified by more undifferentiated biological hallmarks, enhanced expansion and clonal capacity as compared to the more mature, relatively slow-propagating mesenchymal and proneural cells, likely endowed with a higher potential for infiltration either ex vivo or in vivo. Importantly, the combination of DCX and EGFR markers, selectively enriched among GSCs pools, almost exactly predicted GBM patients’ clusters together with their survival and drug response. Conclusions In this study we report that an inherent enrichment of distinct GSCs pools underpin the functional inter-cluster variances displayed by GBM patients. We uncover two selectively represented novel functional biomarkers capable of discriminating GBM patients’ stratification, survival and drug response, setting the stage for the determination of patient-tailored diagnostic and prognostic strategies and, mostly, for the design of appropriate, patient-selective treatment protocols.

Abstract Background Extracellular vesicles (EVs) are membrane-enclosed particles released systemically by all cells, including tumours. Tumour EVs have been shown to manipulate their local environments as well as distal targets to sustain the tumour in a variety of tumours, including glioblastoma (GBM). We have previously demonstrated the dual role of the glial water channel aquaporin-4 (AQP4) protein in glioma progression or suppression depending on its aggregation state. However, its possible role in communication mechanisms in the microenvironment of malignant gliomas remains to be unveiled. Results Here we show that in GBM cells AQP4 is released via EVs that are able to affect the GBM microenvironment. To explore this role, EVs derived from invasive GBM cells expressing AQP4-tetramers or apoptotic GBM cells expressing orthogonal arrays of particles (AQP4-OAPs) were isolated, using a differential ultracentrifugation method, and were added to pre-seeded GBM cells. Confocal microscopy analysis was used to visualize the interaction and uptake of AQP4-containing EVs by recipient cells. Chemoinvasion and Caspase3/7 activation assay, performed on recipient cells after EVs uptake, revealed that EVs produced by AQP4-tetramers expressing cells were able to drive surrounding tumour cells toward the migratory phenotype, whereas EVs produced by AQP4-OAPs expressing cells drive them toward the apoptosis pathway. Conclusion This study demonstrates that the different GBM cell phenotypes can be transferred by AQP4-containing EVs able to influence tumour cell fate toward invasiveness or apoptosis. This study opens a new perspective on the role of AQP4 in the brain tumour microenvironment associated with the EV-dependent communication mechanism. Graphical Abstract

Construction is known as one of the most dangerous industries in terms of worker safety. Collisions due the excessive proximity of workers to moving construction vehicles are one of the leading causes of fatal and non-fatal accidents on construction sites internationally. Proximity warning systems (PWS) have been proposed in the literature as a solution to detect the risk for collision and to alert workers and equipment operators in time to prevent collisions. Although the role of sensing technologies for situational awareness has been recognised in previous studies, several factors still need to be considered. This paper describes the design of a prototype sensor-based PWS, aimed mainly at small and medium-sized construction companies, to collect real-time data directly from construction sites and to warn workers of a potential risk of collision accidents. It considers, in an integrated manner, factors such as cost of deployment, the actual nature of a construction site as an operating environment and data protection. A low-cost, ultra-wideband (UWB)-based proximity detection system has been developed that can operate with or without fixed anchors. In addition, the PWS is compliant with the General Data Protection Regulation (GDPR) of the European Union. A privacy-by-design approach has been adopted and privacy mechanisms have been used for data protection. Future work could evaluate the PWS in real operational conditions and incorporate additional factors for its further development, such as studies on the timely interpretation of data.

Abstract Background Glioblastoma multiforme (GBM) is an incurable tumor, with a median survival rate of only 14–15 months. Along with heterogeneity and unregulated growth, a central matter in dealing with GBMs is cell invasiveness. Thus, improving prognosis requires finding new agents to inhibit key multiple pathways, even simultaneously. A subset of GBM stem-like cells (GSCs) may account for tumorigenicity, representing, through their pathways, the proper cellular target in the therapeutics of glioblastomas. GSCs cells are routinely enriched and expanded due to continuous exposure to specific growth factors, which might alter some of their intrinsic characteristic and hide therapeutically relevant traits. Methods By removing exogenous growth factors stimulation, here we isolated and characterized a subset of GSCs with a “mitogen-independent” phenotype (I-GSCs) from patient’s tumor specimens. Differential side-by-side comparative functional and molecular analyses were performed either in vitro or in vivo on these cells versus their classical growth factor (GF)-dependent counterpart (D-GSCs) as well as their tissue of origin. This was performed to pinpoint the inherent GSCs’ critical regulators, with particular emphasis on those involved in spreading and tumorigenic potential. Transcriptomic fingerprints were pointed out by ANOVA with Benjamini-Hochberg False Discovery Rate (FDR) and association of copy number alterations or somatic mutations was determined by comparing each subgroup with a two-tailed Fisher’s exact test. The combined effects of interacting in vitro and in vivo with two emerging GSCs’ key regulators, such as Wnt5a and EphA2, were then predicted under in vivo experimental settings that are conducive to clinical applications. In vivo comparisons were carried out in mouse-human xenografts GBM model by a hierarchical linear model for repeated measurements and Dunnett’s multiple comparison test with the distribution of survival compared by Kaplan–Meier method. Results Here, we assessed that a subset of GSCs from high-grade gliomas is self-sufficient in the activation of regulatory growth signaling. Furthermore, while constitutively present within the same GBM tissue, these GF-independent GSCs cells were endowed with a distinctive functional and molecular repertoire, defined by highly aggressive Wnt5aHigh/EphA2Low profile, as opposed to Wnt5aLow/EphA2High expression in sibling D-GSCs. Regardless of their GBM subtype of origin, I-GSCs, are endowed with a raised in vivo tumorigenic potential than matched D-GSCs, which were fast-growing ex-vivo but less lethal and invasive in vivo. Also, the malignant I-GSCs’ transcriptomic fingerprint faithfully mirrored the original tumor, bringing into evidence key regulators of invasiveness, angiogenesis and immuno-modulators, which became candidates for glioma diagnostic/prognostic markers and therapeutic targets. Particularly, simultaneously counteracting the activity of the tissue invasive mediator Wnt5a and EphA2 tyrosine kinase receptor addictively hindered GSCs’ tumorigenic and invasive ability, thus increasing survival. Conclusion We show how the preservation of a mitogen-independent phenotype in GSCs plays a central role in determining the exacerbated tumorigenic and high mobility features distinctive of GBM. The exploitation of the I-GSCs' peculiar features shown here offers new ways to identify novel, GSCs-specific effectors, whose modulation can be used in order to identify novel, potential molecular therapeutic targets. Furthermore, we show how the combined use of PepA, the anti-Wnt5a drug, and of ephrinA1-Fc to can hinder GSCs’ lethality in a clinically relevant xenogeneic in vivo model thus being conducive to perspective, novel combinatorial clinical application.

In the architecture, engineering and construction industry, site management during construction is a key phase. Scheduling activities and monitoring their progress allow any deviations from the schedule to be identified so that timely action can be taken. Until now, the monitoring phase has mainly been characterised by inspections in which the construction site manager manually collects data and produces a summary report. This proves to be a time-consuming process and is prone to errors. The authors propose an innovative construction progress monitoring method that combines BIM-based construction scheduling (4D BIM) with periodic geometric surveying using an indoor mobile mapping system (iMMS). Ten surveys were carried out on a real case study, producing point clouds to be compared with the 4D BIM, thereby comparing the as-built with the as-planned. The comparison was carried out using Sitemotion exploiting a custom class, the work breakdown structure (WBS), added to the BIM to associate each element with its scheduled construction date. The results show how the proposed method can effectively support the evaluation of construction progress, allowing the monitoring to be performed digitally and linked to the BIM. The paper details the proposed methodology, highlighting the problems encountered and suggesting adjustments for future implementation.

Abstract Shedding synchrotron light on microfluidic systems, exploring several contrasts in situ/operando at the nanoscale, like X-ray fluorescence, diffraction, luminescence, and absorption, has the potential to reveal new properties and functionalities of materials across diverse areas, such as green energy, photonics, and nanomedicine. In this work, we present the micro-fabrication and characterization of a multifunctional polyester/glass sealed microfluidic device well-suited to combine with analytical X-ray techniques. The device consists of smooth microchannels patterned on glass, where three gold electrodes are deposited into the channels to serve in situ electrochemistry analysis or standard electrical measurements. It has been efficiently sealed through an ultraviolet-sensitive sticker-like layer based on a polyester film, and The burst pressure determined by pumping water through the microchannel(up to 0.22 MPa). Overall, the device has demonstrated exquisite chemical resistance to organic solvents, and its efficiency in the presence of biological samples (proteins) is remarkable. The device potentialities, and its high transparency to X-rays, have been demonstrated by taking advantage of the X-ray nanoprobe Carnaúba/Sirius/LNLS, by obtaining 2D X-ray nanofluorescence maps on the microchannel filled with water and after an electrochemical nucleation reaction. To wrap up, the microfluidic device characterized here has the potential to be employed in standard laboratory experiments as well as in in situ and in vivo analytical experiments using a wide electromagnetic window, from infrared to X-rays, which could serve experiments in many branches of science.

Objective To determine if methotrexate or folic acid prescription was associated with differential risk for COVID-19 diagnosis or mortality.Design Case–control analysis.Setting The population-based UK Biobank (UKBB) cohort.Participants Data from 380 380 UKBB participants with general practice prescription data for 2019–2021. Updated medical information was retrieved on 13 December 2021.Primary and secondary outcome measures The outcomes of COVID-19 diagnosis and COVID-19-related mortality were analysed by multivariable logistic regression. Exposures evaluated were prescription of folic acid and/or methotrexate. Criteria for COVID-19 diagnosis were (1) a positive SARS-CoV-2 test or (2) ICD-10 code for confirmed COVID-19 (U07.1) or probable COVID-19 (U07.2) in hospital records, or death records. By these criteria, 26 003 individuals were identified with COVID-19 of whom 820 were known to have died from COVID-19. Logistic regression statistical models were adjusted for age sex, ethnicity, Townsend deprivation index, body mass index, smoking status, presence of rheumatoid arthritis, sickle cell disease, use of anticonvulsants, statins and iron supplements.Results Compared with people prescribed neither folic acid nor methotrexate, people prescribed folic acid supplementation had increased risk of diagnosis of COVID-19 (OR 1.51 (1.42–1.61)). The prescription of methotrexate with or without folic acid was not associated with COVID-19 diagnosis (p≥0.18). People prescribed folic acid supplementation had positive association with death after a diagnosis of COVID-19 (OR 2.64 (2.15–3.24)) in a fully adjusted model. The prescription of methotrexate in combination with folic acid was not associated with an increased risk for COVID-19-related death (1.07 (0.57–1.98)).Conclusions We report an association of increased risk for COVID-19 diagnosis and COVID-19-related death in people prescribed folic acid supplementation. Our results also suggest that methotrexate might attenuate these associations.

Abstract Gout flare prophylaxis and therapy use in people with underlying chronic kidney disease (CKD) is challenging, given limited treatment options and risk of worsening renal function with inappropriate treatment dosing. This literature review aimed to describe the current literature on the efficacy and safety of gout flare prophylaxis and therapy use in people with CKD stages 3–5. A literature search via PubMed, the Cochrane Library, and EMBASE was performed from 1 January 1959 to 31 January 2018. Inclusion criteria were studies with people with gout and renal impairment (i.e. estimated glomerular filtration rate (eGFR) or creatinine clearance (CrCl)

Abstract Background Colorectal cancer (CRC) harboring BRAF V600E mutation exhibits low response to conventional therapy and poorest prognosis. Due to the emerging correlation between gut microbiota and CRC carcinogenesis, we investigated in serrated BRAF V600E cases the existence of a peculiar fecal microbial fingerprint and specific bacterial markers, which might represent a tool for the development of more effective clinical strategies. Methods By injecting human CRC stem-like cells isolated from BRAF V600E patients in immunocompromised mice, we described a new xenogeneic model of this subtype of CRC. By performing bacterial 16S rRNA sequencing, the fecal microbiota profile was then investigated either in CRC-carrying mice or in a cohort of human CRC subjects. The microbial communities’ functional profile was also predicted. Data were compared with Mann-Whitney U, Welch’s t-test for unequal variances and Kruskal-Wallis test with Benjamini–Hochberg false discovery rate (FDR) correction, extracted as potential BRAF class biomarkers and selected as model features. The obtained mean test prediction scores were subjected to Receiver Operating characteristic (ROC) analysis. To discriminate the BRAF status, a Random Forest classifier (RF) was employed. Results A specific microbial signature distinctive for BRAF status emerged, being the BRAF-mutated cases closer to healthy controls than BRAF wild-type counterpart. In agreement, a considerable score of correlation was also pointed out between bacteria abundance from BRAF-mutated cases and the level of markers distinctive of BRAF V600E pathway, including those involved in inflammation, innate immune response and epithelial-mesenchymal transition. We provide evidence that two candidate bacterial markers, Prevotella enoeca and Ruthenibacterium lactatiformans, more abundant in BRAF V600E and BRAF wild-type subjects respectively, emerged as single factors with the best performance in distinguishing BRAF status (AUROC = 0.72 and 0.74, respectively, 95% confidence interval). Furthermore, the combination of the 10 differentially represented microorganisms between the two groups improved performance in discriminating serrated CRC driven by BRAF mutation from BRAF wild-type CRC cases (AUROC = 0.85, 95% confidence interval, 0.69–1.01). Conclusion Overall, our results suggest that BRAF V600E mutation itself drives a distinctive gut microbiota signature and provide predictive CRC-associated bacterial biomarkers able to discriminate BRAF status in CRC patients and, thus, useful to devise non-invasive patient-selective diagnostic strategies and patient-tailored optimized therapies.

Understanding the tumor microenviroment is important before it can be exploited therapeutically. Here, the authors use single cell sequencing to study stromal cells in mouse tumors and identify a subset of interferon-licensed cancer associated fibroblasts that appear after anti-TGFβ treatment.

Regeneration of existing buildings and abandoned areas is a governmental and European key objective. This research focuses on Poveglia: an abandoned island in the Venetian lagoon, Italy. The underpinning idea concerns the upgrade of Poveglia to a zero energy Cognitive University Campus, connected to other educational institutions in the lagoon. The vision aims at reusing abandoned islands combining architecture, learning environments and digital networks, to demonstrate the potential of new technologies for the transformation of abandoned areas. The case study demonstrates the effectiveness of employing improved and revised systems and tools, developed in eLUX lab at the University of Brescia, for the regeneration of the island. The inclusion of the island in a smart campus network results in triggering positive synergies both enhancing energy efficiency and the learning environment. A further development concerns the setup of strategies for historical preservation, energy efficiency and renewable energy production.

We know Kocher's name as an anatomical reference in neurosurgery. In fact, Theodor Kocher was a Swiss general surgeon, and his contributions were such that Kocher was honored in 1909 with the Nobel Prize in Medicine and Physiology, and he was the first surgeon to receive this honor. Kocher participated in the initial scientific phase of medicine, living with names that are in history, as well as him; Langenbeck and Virchow, Lucke, Billroth, Horsley, Lister, Halstedt, Pasteur, Osler, Lawson Tait, Verneuil, and a long list and other icons of the time. The present account rescues the many important facets and contributions of the Swiss surgeon Theodor Kocher, and his relationship with several of them. Kocher's memory, surgical instruments and literary production are preserved in a small wing of the University of Bern. The present article highlights how intense Kocher's dedication to the medical field was.

Abstract The main objective of this phase I trial was to assess the feasibility and safety of microtransplanting human neural stem cell (hNSC) lines into the spinal cord of patients with amyotrophic lateral sclerosis (ALS). Eighteen patients with a definite diagnosis of ALS received microinjections of hNSCs into the gray matter tracts of the lumbar or cervical spinal cord. Patients were monitored before and after transplantation by clinical, psychological, neuroradiological, and neurophysiological assessment. For up to 60 months after surgery, none of the patients manifested severe adverse effects or increased disease progression because of the treatment. Eleven patients died, and two underwent tracheotomy as a result of the natural history of the disease. We detected a transitory decrease in progression of ALS Functional Rating Scale Revised, starting within the first month after surgery and up to 4 months after transplantation. Our results show that transplantation of hNSC is a safe procedure that causes no major deleterious effects over the short or long term. This study is the first example of medical transplantation of a highly standardized cell drug product, which can be reproducibly and stably expanded ex vivo, comprising hNSC that are not immortalized, and are derived from the forebrain of the same two donors throughout this entire study as well as across future trials. Our experimental design provides benefits in terms of enhancing both intra‐ and interstudy reproducibility and homogeneity. Given the potential therapeutic effects of the hNSCs, our observations support undertaking future phase II clinical studies in which increased cell dosages are studied in larger cohorts of patients. Stem Cells Translational Medicine 2019;8:887&897

Background: Despite their lethality and ensuing clinical and therapeutic relevance, circulating tumor cells (CTCs) from colorectal carcinoma (CRC) remain elusive, poorly characterized biological entities. Methods and findings: We perfected a cell system of stable, primary lines from human CRC showing that they possess the full complement of ex- and in-vivo, in xenogeneic models, characteristics of CRC stem cells (CCSCs). Here we show how tumor-initiating, CCSCs cells can establish faithful orthotopic phenocopies of the original disease, which contain cells that spread into the circulatory system. While in the vascular bed, these cells retain stemness, thus qualifying as circulating CCSCs (cCCSCs). This is followed by the establishment of lesions in distant organs, which also contain resident metastatic CCSCs (mCCSCs). Interpretation: Our results support the concept that throughout all the stages of CRC, stemness is retained as a continuous property by some of their tumor cells. Importantly, we describe a useful standardized model that can enable isolation and stable perpetuation of human CRC's CCSCs, cCCSCs and mCCSCs, providing a useful platform for studies of CRC initiation and progression that is suitable for the discovery of reliable stage-specific biomarkers and the refinement of new patient-tailored therapies. Fund: This work was financially supported by grants from “Ministero della Salute Italiano”(GR-2011-02351534, RC1703IC36 and RC1803IC35) to Elena Binda and from “Associazione Italiana Cancro” (IG-14368) Angelo L. Vescovi. None of the above funders have any role in study design, data collection, data analysis, interpretation, writing the project. Keywords: Human colorectal carcinoma (CRC), Stemness, CRC stem cells, CRC circulating stem cells, CRC metastatic stem cells, CRC biology and biomarkers, Anti-CRC SCs strategies

The new paradigm of smart building requires the accomplishment of occupants needs through the analysis of data gathered within the building, which switches from activities host to provider of customized services for occupants. The digitalization supports this new approach, as the implementation of Building Management Systems (BMS) in Building Information Modelling (BIM) environment allows to link the information collected to a database. The contribution is focused on a case study of the University of Cagliari, the Mandolesi Pavilion, and it is aimed at implementing and improving energy audit procedures by the use of Building Information Modelling.

Abstract Background Many gout comorbidities (e.g., hypertension) are correlated with serum urate. In this investigation, we identified risk factors (e.g., systolic blood pressure [SBP]), that (1) are associated with incident gout, (2) have effects on gout risk that cannot be fully explained by correlated differences in serum urate, and (3) may modulate the relationship between gout and serum urate. Methods Using data from the Atherosclerosis Risk in Communities (ARIC) study, we estimated the unadjusted associations between gout and risk factors by calculating ORs and using chi-square tests. The adjusted associations were analyzed using logistic regression by sequentially adding (1) one risk factor at a time or (2) all risk factors, to a baseline model that includes serum urate only. Stepwise selection was used to select main effects. Two-way interactions of variables from the main effects model were also analyzed. Results Average gout incidence was 2.7 per 1000 people per year. Serum urate was highly associated with incident gout, with odd ratios of 3.16 [95% CI 2.11, 4.76] and 25.9 [95% CI 17.2, 38.4] for moderately high (6–8 mg/dl) and high serum urate (> 8 mg/dl), relative to normal serum urate (

Herein, we explored the impact of the lysosome dysfunction during the progression of Amyotrophic Lateral Sclerosis type-1 (ALS1). We conducted the study in non-neural cells, primary fibroblasts (rFFFs), and bone marrow-mesenchymal stem cells (rBM-MSCs), isolated from the animal model ratG93A for ALS1 at two stages of the disease: Pre-symptomatic-stage (ALS1-PreS) and Terminal-stage (ALS1-EndS). We documented the storage of human mutant Superoxide Dismutase 1, SOD1G93A (SOD1*) in the lysosomes of ALS1-rFFFs and ALS1-rBM-MSCs and demonstrated the hallmarks of the disease in non-neural cells as in ratG93A-ALS1-tissues. We showed that the SOD1* storage is associated with the altered glycohydrolases and proteases levels in tissues and both cell types from ALS1-PreS to ALS1-EndS. Only in ALS1-rFFFs, the lysosomes lost homeostasis, enlarge drastically, and contribute to the cell metabolic damage. Contrariwise, in ALS1-rBM-MSCs, we found a negligible metabolic dysfunction, which makes these cells’ status similar to WT. We addressed this phenomenon to a safety mechanism perhaps associated with an enhanced lysosomal autophagic activity in ALS1-rBM-MSCs compared to ALS1-rFFFs, in which the lysosomal level of LC3-II/LC3I was comparable to that of WT-rFFFs. We suggested that the autophagic machinery could balance the storage of SOD1* aggregates and the lysosomal enzyme dysfunction even in ALS1-EndS-stem cells.

Aspects of terrestrial microgrids and ship power systems are examined. The work exposes a variety of technical synergies from these two power systems to effectively advance their technologies. Understanding their overlap allows congruent efforts to target both systems; understanding their differences hinders conflict and redundancy in early-stage design. The paper concludes by highlighting how an understanding of both systems can reduce the investment in research resources.

Introduction: Road traffic accidents (RTAs) are a major cause of death and disability in South Sudan. The purpose of this study was to investigate whether violation of traffic rules is the main cause of RTAs. Method: A cross sectional study design was used with quantitative data covering January – December 2014. The main objective of the research was to understand the epidemiology of RTAs in order to develop preventive measures. A total of 1725cases from road RTAs data were extracted from the directorate of traffic police Central Equatoria state Juba and Juba teaching hospital. Results: Most (99.5%) of the RTA drivers were not under the influence of alcohol. Most accidents were caused by male drivers (99%). The highest number of RTAs took place in August (11%). Drivers of private vehicles caused most accidents (37%). Most drivers (46%) were aged 20-30 years. RTAs occurred most often on city roads (89.83%). Conclusion: This leads us to conclude that a comprehensive safety system is needed that are premised on the idea of community-based awareness of traffic rules and safety regulations. Resources are limited so there is a need to harness local resources including the local community. More efforts are needed to improve road safety education among the youth/integrate safety into road design.

Sistemas valvulares para controle de hidrocefalia funcionam por meio de diferencial de pressão. As inovações tecnológicas surgidas após a primeira geração de válvulas tentam amenizar o desequilíbrio hidráulico causado pelo desvio artificial do liquor para fora da cavidade intracraniana causado pelo sistema valvular, e que agora sofrem com forças gravitacionais antes compensadas por mecanismos fisiológicos. Este trabalho esclarece ao neurocirurgião os parâmetros que qualificam o nível de desempenho das válvulas frequentemente utilizadas na prática neurocirúrgica, priorizando o entendimento do gráfico “pressão versus vazão”. Para tal, os gráficos foram exemplificados por intermédio do teste de uma válvula de hidrocefalia de primeira geração disponibilizada recentemente, por meio dos testes da ISO 7197.

O fator estimulador de colônias granulocitárias (G-CSF) é uma glicoproteína descrita há mais de vinte anos, e é largamente utilizada para tratamento de estados neutropênicos e no transplante de medula óssea. O G-CSF estimula células-tronco hematopoéticas e regula crucialmente a sobrevivência de neutrófilos maduros, pós-mitóticos, através da inibição da apoptose. Além do efeito sistêmico, mais recentemente tem-se demonstrado uma surpreendente atividade do G-CSF no sistema nervoso central. A administração de G-CSF mobiliza células-tronco e progenitoras da medula óssea para o sangue periférico, que, por sua vez, atravessa a barreira hemato-encefálica (BHE) e se dirige à área acometida do cérebro. A atividade do G-CSF no sistema nervoso central tem sido caracterizada como multimodal, pois, além do efeito mobilizador de células da medula óssea, demonstrou uma ação direta neuroprotetora através de diferentes mecanismos, tais como a atividade antiapoptótica em neurônios, regeneração da vascularização, efeito anti-inflamatório e estimulação da neurogênese endógena. Este relato sumariza a ação do G-CSF no sistema nervoso central e aborda seu potencial para o emprego no acidente vascular cerebral.The granulocyte colony-stimulating-factor (G-CSF) is a glycoproteina which has been described for decades, and it is commonly utilized in the treatment of neutropenic states and bone marrow transplants. G-CSF stimulates hematopoietic stem-cels e crucially regulates the survival of mature neutrophils through a mechanism of apoptosis inhibition. Beyond its systemic effect, recently it has been shown its surprising activity in the central nervous system (CNS). G-CSF administration mobilizes bone marrow stem cells para systemic blood, and those cells cross the blood-brain-barrier e target brain's damaged area. G-CSF's activity in the CNS has been defined as multimodal, because additionally it has been demonstrated a direct neuroprotective action through different mechanisms such as antiapoptotic activity, angiogenesis, anti-inflamatory effect, and stimulation of endogenous neurogenesis. This paper sumarizes G-CSF action in the CNS and approaches its potential para use in stroke.

The presence of drugs and their metabolites in surface waters and municipal effluents has been reported in several studies, but its impacts on aquatic organisms are not yet well understood. This study investigated the effects of acute exposure to the antipsychotic risperidone on the stress and behavioral responses in zebrafish. It became clear that intermediate concentration of risperidone inhibited the hypothalamic-pituitary-interrenal axis and displayed anxiolytic-like effects in zebrafish. The data presented here suggest that the presence of this antipsychotic in aquatic environments can alter neuroendocrine and behavior profiles in zebrafish.

There is controversy about whether serum urate (sUA) predicts future cardiovascular disease (CVD) independently of classical risk factors, and the age at which any prediction starts. We studied the sUA-CVD association among generally healthy adults.CARDIA recruited 5115 black and white individuals aged 18-30 years in 1985-1986 (year-0). Fatal and nonfatal CVD events by year 27 (n = 164) were ascertained during annual contacts and classified using medical records. The association with sUA (year-0, 10, 15 and 20) was modeled using Cox proportional hazards regression, pooling over gender-specific quartiles.Mean sUA concentration was higher in men than women, but increased over time in both genders. Those with elevated sUA had worse metabolic profiles that substantially deteriorated over time. Adjusting for demographic and lifestyle factors (the minimal model), baseline sUA concentration was positively associated with incident CVD (hazard ratio (HR) per mg/dL = 1.21; 95% confidence interval: 1.05, 1.39; P = 0.005). This positive association attenuated to nonsignificance in the full model accounting simultaneously for classical CVD risk factors (HR = 1.09; 0.94, 1.27; P = 0.24). Both the minimal and full models appeared to show stronger associations (than year-0 sUA) between year-10 sUA and incident CVD (HR = 1.27 and 1.12, respectively), but sUA was not statistically significant in the full model. Despite fewer events, year-15 sUA showed a significant sUA-CVD association pattern, with minimal model association magnitude comparable to year-10, and remained significant in the full model (HR = 1.19; 1.02, 1.40; P = 0.03). Hyperuricemia at year-15 strongly predicted CVD risk (HR = 2.11; 1.34, 3.33; P = 0.001), with some attenuation in the full model (HR = 1.68; P = 0.04).sUA may be an early biomarker for CVD in adults entering middle age. The prediction of CVD by sUA appeared to strengthen with aging. The potential complex relation of sUA with deterioration of a cluster of metabolic abnormalities warrants future exploration.

Este projeto é trabalho do Departamento de Trauma da Sociedade Brasileira de Neurocirurgia, realizado com a colaboração de diversas instituições, com o objetivo de conceber uma infra-estrutura que permita estudos prospectivos (multicêntricos ou não) de questões relacionadas à neurotraumatologia, coletando dados via Internet entre instituições. Descrevemos a situação atual do “Projeto Diretrizes de Atendimento ao Traumatismo Craniencefálico”. Apresentamos os resultados iniciais de um estudo cooperativo entre diversas instituições médicas através da criação de um banco de dados e estabelecemos um novo protocolo de estudo. Propomos que o sistema atual evolua à semelhança do BrainIT Group. No momento é uma proposta conceitual, de uma estrutura de coordenação entre serviços e de acesso a bancos de dados e que estabeleça critérios para publicação.

Objetivo: a hiperdrenagem devido a acidentes e manipulação inadequada de sistemas de drenagem externa é um fato e submete o paciente a conseqüências graves, muitas vezes até à morte. Essa complicação mecânica tem sido subestimada e para a qual não existe mecanismo protetor até esse momento. Este trabalho procura responder às seguintes questões: (1) qual é o tempo necessário para que ocorra a drenagem de todo o líquor contido nos ventrículos para o sistema de drenagem externa? (2) qual é a influência do gradiente hidrostático no tempo de drenagem? (3) qual a influência da pressão intraventricular no tempo de drenagem? Materiais e métodos: utilizando-se de uma bancada de testes para hidrodinâmica, as tubagens de três sistemas, comerciais disponíveis no país, foram testadas quanto às suas características de fluxo. Foram simulados gradientes hidrostáticos negativos de 12,5; 25; 50 e 100 cm, e pressão intraventricular (PIV) de 5, 10, 15 e 20 cm H2O. Resultados: o tempo de escoamento do líquor ventricular depende do raio e do comprimento do tubo do sistema utilizado. Entretanto, qualquer que seja o sistema utilizado, o escoamento ocorre muito rapidamente. O gradiente hidrostático é o fator mais importante na drenagem do conteúdo ventricular no efeito sifão; a PIV e o raio do tubo têm importâncias relativas, que se atenuam conforme aumenta o gradiente hidrostático negativo. Conclusão: a quantificação temporal do fluxo livre de uma bolsa de drenagem ressalta a importância clínica de se desenvolver mecanismos protetores contra o efeito sifão em sistemas de drenagem externa.

Este artigo é um estudo laboratorial das características do fluxo e resistência de modelos de vários tipos de cateteres peritoneais, inclusive os disponíveis comercialmente. Utilizou-se uma bancada de testes que permitiu um controle preciso da pressão de perfusão do sistema. Os resultados demonstram que os cateteres comercialmente disponíveis não possuem resistência (Rout) significativa; o cateter aberto apresentou uma Rout que variou de 1,12 a 1,95 mmHg/ml/min, e o cateter com fendas de 10 mm apresentou uma Rout que variou de 1,22 a 1,26 mmHg/ml/min. Em humanos, os níveis considerados normais do elemento resistivo da dinâmica liquórica (Rout) é de até 3 mmHg/ml/min. Isso significa que os cateteres peritoneais testados reproduzem os valores dos elementos resistivos fisiológicos. Contudo, considerando-se a hipótese de Kajimoto, os cateteres de 8 mm e 9 mm possuem potencial para acrescentar o elemento resistivo adequado ao sistema de derivação. O cateter com fendas de 8 mm teve, em ΔPinicial de 5,15 mmHg, um fluxo de 2,11 ml/min e Rout de 2,45 mmHg/ml/min e, em ΔPsentado (14 mmHg), um fluxo de 9,96 ml/min e Rout de 1,41 mmHg/ml/min, representando um elemento resistivo adicional de 25% quando comparado ao cateter aberto tanto em ΔP inicial quanto em ΔPsentado. O cateter com fendas de 9 mm teve, em ΔPinicial de 4,05 mmHg, um fluxo de 1,93 ml/min e Rout de 2,1 mmHg/ml/min, e em ΔPsentado , um fluxo de 10,32 ml/min e Rout de 1,36 mmHg/ml/min, representando um elemento resistivo adicional de 7% em ΔPinicial e de 21% em ΔPsentado quando comparados com o cateter aberto. Assim, os cateteres de 8 mm e 9 mm mostraram potencial para contribuir como um elemento resistivo adicional para limitar o efeito sifão, e merecerão estudos futuros para se observar os efeitos hidrodinâmicos numa bancada de testes que inclua um sistema valvular.

Myofibroblasts (Mfs) that persist in a healing wound promote extracellular matrix (ECM) accumulation and excessive tissue contraction. Increased levels of integrin αvβ5 promote the Mf phenotype and other fibrotic markers. Previously we reported that maintaining uPA (urokinase plasminogen activator) bound to its cell-surface receptor, uPAR prevented TGFβ-induced Mf differentiation. We now demonstrate that uPA/uPAR controls integrin β5 protein levels and in turn, the Mf phenotype. When cell-surface uPA was increased, integrin β5 levels were reduced (61%). In contrast, when uPA/uPAR was silenced, integrin β5 total and cell-surface levels were increased (2-4 fold). Integrin β5 accumulation resulted from a significant decrease in β5 ubiquitination leading to a decrease in the degradation rate of internalized β5. uPA-silencing also induced α-SMA stress fiber organization in cells that were seeded on collagen, increased cell area (1.7 fold), and increased integrin β1 binding to the collagen matrix, with reduced activation of β1. Elevated cell-surface integrin β5 was necessary for these changes after uPA-silencing since blocking αvβ5 function reversed these effects. Our data support a novel mechanism by which downregulation of uPA/uPAR results in increased integrin αvβ5 cell-surface protein levels that regulate the activity of β1 integrins, promoting characteristics of the persistent Mf.

ObjectiveTo determine if methotrexate or folic acid prescription was associated with differential risk for COVID-19 diagnosis or mortality.DesignCase-control analysis.SettingThe population-based UK Biobank (UKBB) cohort.ParticipantsData from 380 380 UKBB participants with general practice prescription data for 2019-2021. Updated medical information was retrieved on 13 December 2021.Primary and secondary outcome measuresThe outcomes of COVID-19 diagnosis and COVID-19-related mortality were analysed by multivariable logistic regression. Exposures evaluated were prescription of folic acid and/or methotrexate. Criteria for COVID-19 diagnosis were (1) a positive SARS-CoV-2 test or (2) ICD-10 code for confirmed COVID-19 (U07.1) or probable COVID-19 (U07.2) in hospital records, or death records. By these criteria, 26 003 individuals were identified with COVID-19 of whom 820 were known to have died from COVID-19. Logistic regression statistical models were adjusted for age sex, ethnicity, Townsend deprivation index, body mass index, smoking status, presence of rheumatoid arthritis, sickle cell disease, use of anticonvulsants, statins and iron supplements.ResultsCompared with people prescribed neither folic acid nor methotrexate, people prescribed folic acid supplementation had increased risk of diagnosis of COVID-19 (OR 1.51 (1.42-1.61)). The prescription of methotrexate with or without folic acid was not associated with COVID-19 diagnosis (p≥0.18). People prescribed folic acid supplementation had positive association with death after a diagnosis of COVID-19 (OR 2.64 (2.15-3.24)) in a fully adjusted model. The prescription of methotrexate in combination with folic acid was not associated with an increased risk for COVID-19-related death (1.07 (0.57-1.98)).ConclusionsWe report an association of increased risk for COVID-19 diagnosis and COVID-19-related death in people prescribed folic acid supplementation. Our results also suggest that methotrexate might attenuate these associations.

The open tank paradigm, also known as novel tank diving test, is a protocol used to evaluate the zebrafish behavior. Several characteristics have been described for this species, including scototaxis, which is the natural preference for dark environments in detriment of bright ones. However, there is no evidence regarding the influence of "natural stimuli" in zebrafish subjected to novelty-based paradigms. In this report, we evaluated the spatio-temporal exploratory activity of the short-fin zebrafish phenotype in the open tank after a short-period confinement into dark/bright environments. A total of 44 animals were individually confined during a 10-min single session into one of three environments: black-painted, white-painted, and transparent cylinders (dark, bright, and transparent groups). Fish were further subjected to the novel tank test and their exploratory profile was recorded during a 15-min trial. The results demonstrated that zebrafish increased their vertical exploratory activity during the first 6-min, where the bright group spent more time and travelled a higher distance in the top area. Interestingly, all behavioral parameters measured for the dark group were similar to the transparent one. These data were confirmed by automated analysis of track and occupancy plots and also demonstrated that zebrafish display a classical homebase formation in the bottom area of the tank. A detailed spatio-temporal study of zebrafish exploratory behavior and the construction of representative ethograms showed that the experimental groups presented significant differences in the first 3-min vs. last 3-min of test. Although the main factors involved in these behavioral responses still remain ambiguous and require further investigation, the current report describes an alternative methodological approach for assessing the zebrafish behavior after a forced exposure to different environments. Additionally, the analysis of ethologically-relevant patterns across time could be a potential phenotyping tool to evaluate the zebrafish exploratory profile in the open tank task.

Cell-based therapy holds great promises for demyelinating diseases. Human-derived fetal and adult oligodendrocyte progenitors (OPC) gave encouraging results in experimental models of dysmyelination but their limited proliferation in vitro and their potential immunogenicity might restrict their use in clinical applications. Virtually unlimited numbers of oligodendroglial cells could be generated from long-term self-renewing human (h)-derived neural stem cells (hNSC). However, robust oligodendrocyte production from hNSC has not been reported so far, indicating the need for improved understanding of the molecular and environmental signals controlling hNSC progression through the oligodendroglial lineage. The aim of this work was to obtain enriched and renewable cultures of hNSC-derived oligodendroglial cells by means of epigenetic manipulation.We report here the generation of large numbers of hNSC-derived oligodendroglial cells by concurrent/sequential in vitro exposure to combinations of growth factors (FGF2, PDGF-AA), neurotrophins (NT3) and hormones (T3). In particular, the combination FGF2+NT3+PDGF-AA resulted in the maintenance and enrichment of an oligodendroglial cell population displaying immature phenotype (i.e., proliferation capacity and expression of PDGFRalpha, Olig1 and Sox10), limited self-renewal and increased migratory activity in vitro. These cells generate large numbers of oligodendroglial progeny at the early stages of maturation, both in vitro and after transplantation in models of CNS demyelination.We describe a reliable method to generate large numbers of oligodendrocytes from a renewable source of somatic, non-immortalized NSC from the human foetal brain. We also provide insights on the mechanisms underlying the pro-oligodendrogenic effect of the treatments in vitro and discuss potential issues responsible for the limited myelinating capacity shown by hNSC-derived oligodendrocytes in vivo.

Neural stem cells (NSCs) represent an optimal tool for studies and therapy of neurodegenerative diseases. We recently established a v-myc immortalized human NSC (IhNSC) line, which retains stem properties comparable to parental cells. Oxygen concentration is one of the most crucial environmental conditions for cell proliferation and differentiation both in vitro and in vivo. In the central nervous system, physiological concentrations of oxygen range from 0.55 to 8% oxygen. In particular, in the in the subventricular zone niche area, it's estimated to be 2.5 to 3%.We investigated in vitro the effects of 1, 2.5, 5, and 20% oxygen concentrations on IhNSCs both during proliferation and differentiation. The highest proliferation rate, evaluated through neurosphere formation assay, was obtained at 2.5 and 5% oxygen, while 1% oxygen was most noxious for cell survival. The differentiation assays showed that the percentages of beta-tubIII+ or MAP2+ neuronal cells and of GalC+ oligodendrocytes were significantly higher at 2.5% compared with 1, 5, or 20% oxygen at 17 days in vitro. Mild hypoxia (2.5 to 5% oxygen) promoted differentiation into neuro-oligodendroglial progenitors as revealed by the higher percentage of MAP2+/Ki67+ and GalC+/Ki67+ residual proliferating progenitors, and enhanced the yield of GABAergic and slightly of glutamatergic neurons compared to 1% and 20% oxygen where a significant percentage of GFAP+/nestin+ cells were still present at 17 days of differentiation.These findings raise the possibility that reduced oxygen levels occurring in neuronal disorders like cerebral ischemia transiently lead to NSC remaining in a state of quiescence. Conversely, mild hypoxia favors NSC proliferation and neuronal and oligodendroglial differentiation, thus providing an important advance and a useful tool for NSC-mediated therapy of ischemic stroke and neurodegenerative diseases like Parkinson's disease, multiple sclerosis, and Alzheimer's disease.

Understanding and harnessing cellular potency are fundamental in biology and are also critical to the future therapeutic use of stem cells. Transcriptome analysis of these pluripotent cells is a first step towards such goals. Starting with sources that include oocytes, blastocysts, and embryonic and adult stem cells, we obtained 249,200 high-quality EST sequences and clustered them with public sequences to produce an index of approximately 30,000 total mouse genes that includes 977 previously unidentified genes. Analysis of gene expression levels by EST frequency identifies genes that characterize preimplantation embryos, embryonic stem cells, and adult stem cells, thus providing potential markers as well as clues to the functional features of these cells. Principal component analysis identified a set of 88 genes whose average expression levels decrease from oocytes to blastocysts, stem cells, postimplantation embryos, and finally to newborn tissues. This can be a first step towards a possible definition of a molecular scale of cellular potency. The sequences and cDNA clones recovered in this work provide a comprehensive resource for genes functioning in early mouse embryos and stem cells. The nonrestricted community access to the resource can accelerate a wide range of research, particularly in reproductive and regenerative medicine.