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IntroductionParkinson’s disease (PD) is characterized by an alteration of the walking gait, frequently including a slower self-selected walking speed (SSWS). Although the reduction of walking speed is inherent to people with PD, such speed reduction also represents a potential confounding factor that might partly explain the observed gait differences between PD and control participants.MethodsIn this study, each participant walked along a 25 m level corridor during which vertical ground reaction force signals were recorded using shoes equipped with eight pressure sensors. Vertical ground reaction force signals (using statistical parametric mapping) and temporal and kinetic variables as well as their related variability and asymmetry (using Student’s t-test) were compared between PD (n = 54) and walking-speed-matched control subjects (n = 39).ResultsStatistical parametric mapping did not yield significant differences between PD and control groups for the vertical ground reaction force signal along the walking stance phase. Stride time and single support time (equivalent to swing time) were shorter and peak vertical ground reaction force was larger in PD patients compared to controls (p ≤ 0.05). However, the single support time was no longer different between people with PD and healthy subjects when expressed relatively to stride time (p = 0.07). While single support, double support, and stance times were significantly more variable and asymmetric for PD than for the control group (p ≤ 0.05), stride time was similar (p ≥ 0.07).DiscussionThese results indicate that at matched SSWS, PD patients adopt a higher cadence than control participants. Moreover, the temporal subdivision of the walking gait of people with PD is similar to healthy individuals but the coordination during the double support phase is different. Hence, this study indicates that isolating the speed factor is crucial in gait analysis for PD.

After several field campaigns between 2007 and 2018 in the northwestern region of the Dominican Republic, more than 300 archaeological sites have been registered and revisited. While several of these sites were identified through the scatter of surface material culture, others show terrain modifications in the form of anthropogenic mounds and levelled areas. Researchers have gathered valuable information regarding these features’ functionality and construction processes through large-scale excavations in archaeological sites with anthropogenic mounds, paleoenvironmental studies and remote sensing analyses. These anthropogenic mounds represent a long-term process of formation and were used for a variety of purposes ranging from agricultural to ritual activities. While excavations and small-scale remote sensing can provide a myriad of data to improve our understanding of these archaeological sites, a regional perspective is needed to map the relationship among archaeological sites with and without terrain modifications, to better understand the Indigenous cultural patterns in their regional environment. In this regard, the primary objective of this paper is to explore to what extent these archaeological sites were related to the environment and each other. This was achieved by correlating archaeological data with a set of archaeologically recognized important environmental variables using advanced spatial statistics. The results provide important insights to understand the underlying pattern of archaeological sites in this region and its relationship with the environmental setting.

Background: In vitro and toxicological studies have shown that non-persistent environmental chemicals can perturb thyroid hormone homeostasis. Epidemiological studies with improved exposure assessment (i.e., repeated urine samples) are needed to evaluate effects of these compounds, individually or as a mixture, in humans. We studied the associations between prenatal exposure to non-persistent environmental chemicals and neonatal thyroid hormones. Methods: The study population consisted of 442 mother–child pairs from the French SEPAGES mother–child cohort recruited between July 2014 and July 2017. For each participant, four parabens, five bisphenols, triclosan, triclocarban, benzophenone-3 as well as metabolites of phthalates and of di(isononyl)cyclohexane-1,2-dicarboxylate were assessed in two pools of repeated urine samples (median: 21 spot urines per pool), collected in the 2nd and 3rd trimesters of pregnancy, respectively. Thyroid stimulating hormone (TSH) and total thyroxine (T4) levels were determined in newborns from a heel-prick blood spot. Maternal iodine and selenium were assessed in urine and serum, respectively. Adjusted linear regression (uni-pollutant model) and Bayesian Kernel Machine Regression (BKMR, mixture model) were applied to study overall and sex-stratified associations between chemicals and hormone concentrations. Results: Interaction with child sex was detected for several compounds. Triclosan, three parabens, and one phthalate metabolite (OH-MPHP) were negatively associated with T4 among girls in the uni-pollutant model. BKMR also suggested a negative association between the mixture and T4 in girls, whereas in boys the association was positive. The mixture was not linked to TSH levels, and for this hormone the uni-pollutant model revealed associations with only a few compounds. Conclusion: Our study, based on repeated urine samples to assess exposure, showed that prenatal exposure to some phenols and phthalates disturb thyroid hormone homeostasis at birth. Furthermore, both uni-pollutant and mixture models, suggested effect modification by child sex, while, to date underlying mechanisms for such sex-differences are not well understood.

Introduction Repeated sprints exercise (RSE) performed in hypoxia (RSH) induce greater performance improvement than in normoxia (Brocherie et al., 2017). It has been previously argued that RSH efficiency depend on the oxidative-glycolytic balance which is influenced by sprint duration and exercise-to-rest-ratio (E:R). Indeed, we recently showed that long sprint duration (e.g. 20 s vs 10 s or 5 s) blunts the additional impact of hypoxia on RSH with a constant large E:R of 1:22. However, E:R also influence acute response to RSH. Therefore, this study aims to compare acute responses during RSE to exhaustion with a short E:R (1:6) in normoxia (RSN) and hypoxia (FiO2 = 0.13) and the same sprint durations (5, 10 or 20 s) previously used. Methods On separate visits, 10 active participants completed in random order three RSH and three RSN sessions to exhaustion on a cycle-ergometer (Excalibur Sport, Lode) with three sprint durations and a constant short E:R of 1:6 (5:30; 10:60 and 20:120). Vastus lateralis muscle de-reoxygenation (Oxymon, Artinis Medical Systems) and power output were continuously recorded. Lower limb and breathing discomfort, blood lactate, and ratings of perceived exertion were evaluated immediately after exhaustion. Results Number of sprints and peak power output were higher while blood lactate was lower (all p < 0.001) during 5:30 compared to 10:60 or 20:120. No condition or interaction effects were reported for blood lactate and exercise-related sensation. Blood lactate and muscle deoxyhemoglobin increase (p < 0.001) and total hemoglobin decrease (p = 0.002) during sprint were more pronounced when sprint duration increased (no hypoxia effect). Discussion Similar deoxygenation changes during recovery were reported in hypoxia compared to normoxia suggesting that 1:6 ratio and the associated long recovery period seemed to allow myoglobin O2 store restauration, even with a FiO2 = 13%. Sprint effort should be interspersed by incomplete recovery making impossible the full O2 reloading myoglobin and PCr resynthesis. The increase in blood lactate and the concomitant decrease in muscle oxygenation during sprint with longer duration confirmed a switch in the oxidative-glycolytic balance for energy supply. Since short exercise-to-rest ratio blunt hypoxic effect during RSH due to too long recovery period, hypoxia did not participate to the switch from oxidative to glycolytic energy supply. Conclusion During RSE to exhaustion, a short exercise-to-rest ratio (i.e., 1:6) blunts the specific psychophysiological responses induced by hypoxia, probably due to too long of recovery period. Manipulating effort duration rather than hypoxic exposure is preferable to alter both anaerobic energy system solicitation and power produced during repeated cycling sprints to exhaustion using a short exercise-to-rest ratio. References Brocherie, F., Girard, O., Faiss, R., & Millet, G. P. (2017). Effects of repeated-sprint training in hypoxia on sea-level performance: A meta-analysis. Sports Medicine, 47, 1651-1660. https://doi.org/10.1007/s40279-017-0685-3

In memoriam of Prof. Stan Fakan

Abstract Background Chloride intracellular channel-1 (CLIC1) activity controls glioblastoma proliferation. Metformin exerts antitumor effects in glioblastoma stem cells (GSCs) inhibiting CLIC1 activity, but its low potency hampers its translation in clinical settings. Methods We synthesized a small library of novel biguanide-based compounds that were tested as antiproliferative agents for GSCs derived from human glioblastomas, in vitro using 2D and 3D cultures and in vivo in the zebrafish model. Compounds were compared to metformin for both potency and efficacy in the inhibition of GSC proliferation in vitro (MTT, Trypan blue exclusion assays, and EdU labeling) and in vivo (zebrafish model), migration (Boyden chamber assay), invasiveness (Matrigel invasion assay), self-renewal (spherogenesis assay), and CLIC1 activity (electrophysiology recordings), as well as for the absence of off-target toxicity (effects on normal stem cells and toxicity for zebrafish and chick embryos). Results We identified Q48 and Q54 as two novel CLIC1 blockers, characterized by higher antiproliferative potency than metformin in vitro, in both GSC 2D cultures and 3D spheroids. Q48 and Q54 also impaired GSC self-renewal, migration and invasion, and displayed low systemic in vivo toxicity. Q54 reduced in vivo proliferation of GSCs xenotransplanted in zebrafish hindbrain. Target specificity was confirmed by recombinant CLIC1 binding experiments using microscale thermophoresis approach. Finally, we characterized GSCs from GBMs spontaneously expressing low CLIC1 protein, demonstrating their ability to grow in vivo and to retain stem-like phenotype and functional features in vitro. In these GSCs, Q48 and Q54 displayed reduced potency and efficacy as antiproliferative agents as compared to high CLIC1-expressing tumors. However, in 3D cultures, metformin and Q48 (but not Q54) inhibited proliferation, which was dependent on the inhibition dihydrofolate reductase activity. Conclusions These data highlight that, while CLIC1 is dispensable for the development of a subset of glioblastomas, it acts as a booster of proliferation in the majority of these tumors and its functional expression is required for biguanide antitumor class-effects. In particular, the biguanide-based derivatives Q48 and Q54, represent the leads to develop novel compounds endowed with better pharmacological profiles than metformin, to act as CLIC1-blockers for the treatment of CLIC1-expressing glioblastomas, in a precision medicine approach.

Neuronal PAS domain protein 2 (NPAS2) is a hemeprotein comprising a basic helix–loop–helix domain (bHLH) and two heme-binding sites, the PAS-A and PAS-B domains. This protein acts as a pyridine nucleotide-dependent and gas-responsive CO-dependent transcription factor and is encoded by a gene whose expression fluctuates with circadian rhythmicity. NPAS2 is a core cog of the molecular clockwork and plays a regulatory role on metabolic pathways, is important for the function of the central nervous system in mammals, and is involved in carcinogenesis as well as in normal biological functions and processes, such as cardiovascular function and wound healing. We reviewed the scientific literature addressing the various facets of NPAS2 and framing this gene/protein in several and very different research and clinical fields.

An updated checklist of Mozambique’s vascular plants is presented. It was compiled referring to several information sources such as existing literature, relevant online databases and herbaria collections. The checklist includes 7,099 taxa (5,957 species, 605 subspecies, 537 varieties), belonging to 226 families and 1,746 genera. There are 6,804 angiosperms, 257 pteridophytes, and 38 gymnosperms. A total of 6,171 taxa are native to Mozambique, while 602 are introduced and the remaining 326 taxa were considered as uncertain status. The endemism level for Mozambique’s flora was assessed at 9.59%, including 278 strict-endemic taxa and 403 near-endemic. 58.2% of taxa are herbaceous, while shrubs and trees account respectively for 26.5% and 9.2% of the taxa. The checklist also includes ferns (3.6%), lianas (1.7%), subshrubs (0.5%) and cycads (0.3%). Fabaceae, Poaceae and Asteraceae are the three most represented families, with 891, 543 and 428 taxa, respectively. The extinction risk of 1,667 taxa is included, with 158 taxa listed as Vulnerable, 119 as Endangered and as 24 Critically Endangered. The geographical distribution, known vernacular names and plants traditional uses are also recorded.

Ancylostoma caninum is the most prevalent nematode parasite of dogs. We confirmed multiple-drug resistance (MDR) in several A. caninum isolates to all anthelmintic drug classes approved for the treatment of hookworms in dogs in the USA. Cases of MDR hookworms appear to be highly overrepresented in greyhounds. The aims of this study were to evaluate the drug-resistant phenotypes and genotypes of the A. caninum infecting greyhounds. Fecal samples from greyhounds of the USA were acquired from two greyhound adoption kennels, one active greyhound racing kennel, and three veterinary practices. Fecal egg counts (FECs) were performed on fecal samples from 219 greyhounds, and despite treatment with anthelmintics, the mean FEC was 822.4 eggs per gram (EPG). Resistance to benzimidazoles and macrocyclic lactones were measured using the egg hatch assay (EHA) and the larval development assay (LDA), respectively. We performed 23 EHA and 22 LDA on either individual or pooled feces, representing 54 animals. Mean and median IC50 and IC95 values for the EHA were 5.3 μM, 3.6 μM, and 24.5 μM, 23.4 μM, respectively. For the LDA, the median IC50 value was >1000 nM. These values ranged 62–81 times higher than our susceptible laboratory isolate. Only post-treatment samples were available. For samples collected TAC) resistance polymorphism in 99% of these samples. These clinical, in vitro, and genetic data provide strong evidence that greyhound dogs in the USA are infected with MDR A. caninum at very high levels in prevalence and infection intensity.

In this article, we look at twirling baton clubs characterized by a specific sociality which is part of a life form. This was threatened by the demands of sportivisation from the sport institution. The protests expressed by the clubs show the attachment to a logic of care with the aim of protecting the vulnerabilities and the singular identities of the athletes who are mostly young girls.

The development of novel nanoconstructs for biomedical applications requires the assessment of their biodistribution, metabolism and clearance in single cells, organs and entire organisms in a living environment. To reduce the number of in vivo experiments performed and to refine the methods used, in accordance with the 3Rs principle, this work proposes an ex vivo experimental system to monitor, using fluorescence microscopy, the distribution of nanoparticles in explanted murine skeletal muscle maintained in a bioreactor that can preserve the structural and functional features of the organ for long periods of time. Fluorescently-labelled liposomes and poly(lactide-co-glycolide) (PLGA)-based nanoparticles were injected into the intact soleus muscle (in the distal region close to the tendon) immediately after explants, and their distribution was analysed at increasing incubation times in cross cryosections from the proximal region of the belly. Both nanocarriers were clearly recognized in the muscle and were found to enter and migrate inside the myofibres, whereas their migration in the connective tissue seemed to be limited. In addition, some fluorescent signals were observed inside the macrophages, demonstrating the physiological clearance of the nanocarriers that did not enter the myofibres. Our ex vivo system therefore provides more information than previous in vitro experiments on cultured muscle cells, highlighting the need for the appropriate functionalization of nanocarriers if myofibre targeting is to be improved.

Duty factor (DF) and step frequency (SF) were previously defined as the key running pattern determinants. Hence, this study aimed to investigate the association of DF and SF on 1) the vertical and fore-aft ground reaction force signals using statistical parametric mapping; 2) the force related variables (peaks, loading rates, impulses); and 3) the spring-mass characteristics of the lower limb, assessed by computing the force-length relationship and leg stiffness, for treadmill runs at several endurance running speeds. One hundred and fifteen runners ran at 9, 11, and 13 km/h. Force data (1000 Hz) and whole-body three-dimensional kinematics (200 Hz) were acquired by an instrumented treadmill and optoelectronic system, respectively. Both lower DF and SF led to larger vertical and fore-aft ground reaction force fluctuations, but to a lower extent for SF than for DF. Besides, the linearity of the force-length relationship during the leg compression decreased with increasing DF or with decreasing SF but did not change during the leg decompression. These findings showed that the lower the DF and the higher the SF, the more the runner relies on the optimization of the spring-mass model, whereas the higher the DF and the lower the SF, the more the runner promotes forward propulsion.

Nanoconstructs intended to be used as biomedical tool must be assessed for their capability to cross biological barriers. However, studying in vivo the permeability of biological barriers to nanoparticles is quite difficult due to the many structural and functional factors involved. Therefore, the in vitro modeling of biological barriers -2D cell monocultures, 2D/3D cell co-cultures, microfluidic devices- is gaining more and more relevance in nanomedical research. Microscopy techniques play a crucial role in these studies, as they allow both visualizing nanoparticles inside the biological barrier and evaluating their impact on the barrier components. This paper provides an overview of the various microscopical approaches used to investigate nanoparticle translocation through in vitro biological barrier models. The high number of scientific articles reported highlights the great contribution of the morphological and histochemical approach to the knowledge of the dynamic interactions between nanoconstructs and the living environment.

BACKGROUND:Tumor-associated macrophages (TAMs) are abundant in gliomas and immunosuppressive TAMs are a barrier to emerging immunotherapies. It is unknown to what extent macrophages derived from peripheral blood adopt the phenotype of brain-resident microglia in pre-treatment gliomas. The relative proportions of blood-derived macrophages and microglia have been poorly quantified in clinical samples due to a paucity of markers that distinguish these cell types in malignant tissue. RESULTS:We perform single-cell RNA-sequencing of human gliomas and identify phenotypic differences in TAMs of distinct lineages. We isolate TAMs from patient biopsies and compare them with macrophages from non-malignant human tissue, glioma atlases, and murine glioma models. We present a novel signature that distinguishes TAMs by ontogeny in human gliomas. Blood-derived TAMs upregulate immunosuppressive cytokines and show an altered metabolism compared to microglial TAMs. They are also enriched in perivascular and necrotic regions. The gene signature of blood-derived TAMs, but not microglial TAMs, correlates with significantly inferior survival in low-grade glioma. Surprisingly, TAMs frequently co-express canonical pro-inflammatory (M1) and alternatively activated (M2) genes in individual cells. CONCLUSIONS:We conclude that blood-derived TAMs significantly infiltrate pre-treatment gliomas, to a degree that varies by glioma subtype and tumor compartment. Blood-derived TAMs do not universally conform to the phenotype of microglia, but preferentially express immunosuppressive cytokines and show an altered metabolism. Our results argue against status quo therapeutic strategies that target TAMs indiscriminately and in favor of strategies that specifically target immunosuppressive blood-derived TAMs.

Introdução: Com a pandemia de COVID-19 o Serviço de Controle de Infecções Hospitalares (SCIH) além do desafio em lidar com os fluxos relacionados a COVID-19 e orientações de prevenção para pacientes e colaboradores, também enfrentou a disseminação de microrganismos multirresistentes causadoras de infecções relacionadas à assistência à saúde (IRAS). Objetivo: Avaliar o impacto das ações adotadas na redução de microrganismos multirresistentes (MDR) e no consumo de antimicrobianos (ATM) nas unidades de terapia intensiva (UTIs) durante a pandemia de COVID-19. Método: As ações ocorreram em duas UTIs destinadas a pacientes com diagnóstico de covid-19 de um hospital de nível terciário de assistência à saúde. Foram avaliadas a densidade de incidência de MDR da instituição no período pandêmico e seu comportamento durante a segunda onda, a partir de fevereiro/2021. A intensificação das medidas de controle ocorreu entre 01/06 e 31/08/2021 e incluíram treinamentos voltados à higiene das mãos, uso correto de EPI, monitoramento da higiene ambiental, auditoria por 4 semanas da higiene terminal das UTIs (n = 9), com aplicação de marcadores fluorescentes em 259 pontos definidos pelo SCIH, instituição do banho diário com clorexidina degermante 2% (n = 116), coletas semanal de swab anal avaliação de colonização por MDR (n = 169) e intensificação diária do Programa de Antimicrobial Stewardship nas UTIs. Resultados: Em 2020, a densidade de infecções por K. pneumoniae foi de 1,1 por 1.000 pacientes/dia; P. aeruginosa zero e A. baumannii 0,1, com significativo crescimento entre fevereiro e julho/2021: 2,3; 2,1 e 2,6, por 1.000 pacientes/dia. A auditoria da higiene ambiental da UTI evidenciou 56% de adesão global, enquanto a análise dos pontos individualizados apresentou efetividade de 71%. A adesão dos profissionais das UTI aos treinamentos ministrados foi de 90%. A dose diária definida de antibióticos das UTIs passou de 3.528 para 1.721, após intervenção direta do infectologista nas UTI COVID-19. A colonização por KPC isolada em swab anal (pesquisa de colonização) foi de 7% em junho, 5,6% em julho e 0% em agosto. A colonização por enterococo resistente à vancomicina foi de 24,5% em junho, 16,9% julho e 3,7% agosto. Não identificamos MDR nas infecções notificadas em agosto, novembro e dezembro/2021, setembro 8,2%, outubro 7,5%. Conclusão: As ações de prevenção de infecção e o Programa de Antimicrobial Stewardship tiveram importante impacto para a redução dos MDR e do consumo de ATM nas UTIs de internação de pacientes com COVID-19.

Glioblastoma multiforme (GBM) has high mortality and recurrence rates. Malignancy resilience is ascribed to Glioblastoma Stem Cells (GSCs), which are resistant to Temozolomide (TMZ), the gold standard for GBM post-surgical treatment. However, Nitric Oxide (NO) has demonstrated anti-cancer efficacy in GBM cells, but its potential impact on GSCs remains unexplored. Accordingly, we investigated the effects of NO, both alone and in combination with TMZ, on patient-derived GSCs. Experimentally selected concentrations of diethylenetriamine/NO adduct and TMZ were used through a time course up to 21 days of treatment, to evaluate GSC proliferation and death, functional recovery, and apoptosis. Immunofluorescence and Western blot analyses revealed treatment-induced effects in cell cycle and DNA damage occurrence and repair. Our results showed that NO impairs self-renewal, disrupts cell-cycle progression, and expands the quiescent cells’ population. Consistently, NO triggered a significant but tolerated level of DNA damage, but not apoptosis. Interestingly, NO/TMZ cotreatment further inhibited cell cycle progression, augmented G0 cells, induced cell death, but also enhanced DNA damage repair activity. These findings suggest that, although NO administration does not eliminate GSCs, it stunts their proliferation, and makes cells susceptible to TMZ. The resulting cytostatic effect may potentially allow long-term control over the GSCs’ subpopulation.

Down syndrome (DS) is a genetically-based disease based on the trisomy of chromosome 21 (Hsa21). DS is characterized by intellectual disability in association with several pathological traits among which early aging and altered motor coordination are prominent. Physical training or passive exercise were found to be useful in counteracting motor impairment in DS subjects. In this study we used the Ts65Dn mouse, a widely accepted animal model of DS, to investigate the ultrastructural architecture of the medullary motor neuron cell nucleus taken as marker of the cell functional state. Using transmission electron microscopy, ultrastructural morphometry, and immunocytochemistry we carried out a detailed investigation of possible trisomy-related alteration(s) of nuclear constituents, which are known to vary their amount and distribution as a function of nuclear activity, as well as the effect of adapted physical training upon them. Results demonstrated that trisomy per se affects nuclear constituents to a limited extent; however, adapted physical training is able to chronically stimulate pre-mRNA transcription and processing activity in motor neuron nuclei of trisomic mice, although to a lesser extent than in their euploid mates. These findings are a step towards understanding the mechanisms underlying the positive effect of physical activity in DS.

The home can be a complex environment to understand, as well as to model and predict, due to inherent variability between people’s routines and practices. A one-size-fits-all approach does not consider people’s contextual and institutional influences that contribute to their daily routines. These contextual and institutional factors relate to the household structure and relationship between occupants, as well as the working lifestyle of the occupants. One household can consume resources and live quite differently compared to a similar size household with the same number of occupants due to these factors. Predictive analysis of consumption data can identify this difference to create household-specific modelling to predict occupant routines and practices. Using post-occupancy data from the Fairwater Living Laboratory in Sydney that monitored 39 homes built in a green-star community, this research has utilised machine learning approaches and a K-Means clustering method complemented by t-distributed Stochastic Neighbour Embedding (t-SNE) to show how households follow different daily routines and activities resulting in resource consumption. This analysis has identified energy usage patterns and household groupings with each group following similar daily routines and consumption. The comparison between modelling the precinct as a whole and modelling households individually shows how detail can be lost when aggregating household data at a precinct/community level. This detail can explain why policies or technologies are not as effective as their design due to ignoring the delicate aspects of household routines and practices. These household groupings can provide insight for policymakers to help them understand the different profiles that may be present in the community. These findings are useful for net-zero developments and decarbonization of the built environment through modelling occupant behaviour accurately and developing policies and technologies to suit.

The mild oxidative stress induced by low doses of gaseous ozone (O3) activates the antioxidant cell response through the nuclear factor erythroid 2-related factor 2 (Nrf2), thus inducing beneficial effects without cell damage. Mitochondria are sensitive to mild oxidative stress and represent a susceptible O3 target. In this in vitro study, we investigated the mitochondrial response to low O3 doses in the immortalized, non-tumoral muscle C2C12 cells; a multimodal approach including fluorescence microscopy, transmission electron microscopy and biochemistry was used. Results demonstrated that mitochondrial features are finely tuned by low O3 doses. The O3 concentration of 10 μg maintained normal levels of mitochondria-associated Nrf2, promoted the mitochondrial increase of size and cristae extension, reduced cellular reactive oxygen species (ROS) and prevented cell death. Conversely, in 20 μg O3-treated cells, where the association of Nrf2 with the mitochondria drastically dropped, mitochondria underwent more significant swelling, and ROS and cell death increased. This study, therefore, adds original evidence for the involvement of Nrf2 in the dose-dependent response to low O3 concentrations not only as an Antioxidant Response Elements (ARE) gene activator but also as a regulatory/protective factor of mitochondrial function.

The human genome produces thousands of long noncoding RNAs (lncRNAs)-transcripts >200 nucleotides long that do not encode proteins. Although critical roles in normal biology and disease have been revealed for a subset of lncRNAs, the function of the vast majority remains untested. We developed a CRISPR interference (CRISPRi) platform targeting 16,401 lncRNA loci in seven diverse cell lines, including six transformed cell lines and human induced pluripotent stem cells (iPSCs). Large-scale screening identified 499 lncRNA loci required for robust cellular growth, of which 89% showed growth-modifying function exclusively in one cell type. We further found that lncRNA knockdown can perturb complex transcriptional networks in a cell type-specific manner. These data underscore the functional importance and cell type specificity of many lncRNAs.

Cardiomyocyte renewal represents an unmet clinical need for cardiac regeneration. Stem cell paracrine therapy has attracted increasing attention to resurge rescue mechanisms within the heart. We previously characterized the paracrine effects that human amniotic fluid–derived stem cells (hAFSC) can exert to provide cardioprotection and enhance cardiac repair in preclinical models of myocardial ischemia and cardiotoxicity. Here, we analyze whether hAFSC secretome formulations, namely, hAFSC conditioned medium (hAFSC-CM) over extracellular vesicles (hAFSC-EVs) separated from it, can induce cardiomyocyte renewal. c-KIT+ hAFSC were obtained by leftover samples of II trimester prenatal amniocentesis (fetal hAFSC) and from clinical waste III trimester amniotic fluid during scheduled C-section procedures (perinatal hAFSC). hAFSC were primed under 1% O2 to enrich hAFSC-CM and EVs with cardioactive factors. Neonatal mouse ventricular cardiomyocytes (mNVCM) were isolated from cardiac tissue of R26pFUCCI2 mice with cell cycle fluorescent tagging by mutually exclusive nuclear signal. mNVCM were stimulated by fetal versus perinatal hAFSC-CM and hAFSC-EVs to identify the most promising formulation for in vivo assessment in a R26pFUCCI2 neonatal mouse model of myocardial infarction (MI) via intraperitoneal delivery. While the perinatal hAFSC secretome did not provide any significant cardiogenic effect, fetal hAFSC-EVs significantly sustained mNVCM transition from S to M phase by 2-fold, while triggering cytokinesis by 4.5-fold over vehicle-treated cells. Treated mNVCM showed disorganized expression of cardiac alpha-actinin, suggesting cytoskeletal re-arrangements prior to cell renewal, with a 40% significant downregulation of Cofilin-2 and a positive trend of polymerized F-Actin. Fetal hAFSC-EVs increased cardiomyocyte cell cycle progression by 1.8-fold in the 4-day-old neonatal left ventricle myocardium short term after MI; however, such effect was lost at the later stage. Fetal hAFSC-EVs were enriched with a short isoform of Agrin, a mediator of neonatal heart regeneration acting by YAP-related signaling; yet in vitro application of YAP inhibitor verteporfin partially affected EV paracrine stimulation on mNVCM. EVs secreted by developmentally juvenile fetal hAFSC can support cardiomyocyte renewal to some extension, via intercellular conveyance of candidates possibly involving Agrin in combination with other factors. These perinatal derivative promising cardiogenic effects need further investigation to define their specific mechanism of action and enhance their potential translation into therapeutic opportunity.

O objetivo desse trabalho consistiu em gerar uma tipologia de classificação para os municípios da Amazônia Legal com base em indicadores socioeconômicos e demográficos capazes de representar adequadamente as características da região, possibilitando a construção de uma base territorial de grupos relativamente homogêneos, objetivando, mais particularmente, verificar se era possível identificação de um padrão convergente de municípios convergentes em desmatamento e outros indicadores associados. Foram utilizados 762 municípios que fizeram parte do Projeto de Estimativa do Desmatamento da Amazônia (PRODES) no ano de 2010 e uma base de dados com 19 variáveis abrangendo aspectos econômicos, demográficos, sociais e de posse da terra. Os dados foram submetidos aos métodos multivariados: análise fatorial e análise de agrupamento cluster. Os resultados indicaram como resultado cinco fatores de significado, sendo eles: grau de desenvolvimento médio, desmatamento e seus condicionantes, abertura ao comércio, conservação e direito de propriedades. Os municípios foram, então, agrupados de acordo com os níveis de similaridade, em quatro categorias. Entre os resultados encontrados, o grupo dois destacou-se por ser aquele que mais contém municípios associados a altos valores de desmatamento no ano de 2010.

This article reviews several software programs designed to conduct item response theory (IRT) scale linking and equating. The programs reviewed include IRTEQ, STUIRT, and POLYEQUATE. Features and functionalities of each program are discussed and an example analysis using the common-item non-equivalent groups design in IRTEQ is provided.

Skin is a major administration route for drugs, and all transdermal formulations must be tested for their capability to overcome the cutaneous barrier. Therefore, developing highly reliable skin models is crucial for preclinical studies. The current in vitro models are unable to replicate the living skin in all its complexity; thus, to date, excised human skin is considered the gold standard for in vitro permeation studies. However, skin explants have a limited life span. In an attempt to overcome this problem, we used an innovative bioreactor that allowed us to achieve good structural and functional preservation in vitro of explanted human skin for up to 72 h. This device was then used to set up an in vitro inflammatory model by applying two distinct agents mimicking either exogenous or endogenous stimuli: i.e., dithranol, inducing the contact dermatitis phenotype, and the substance P, mimicking neurogenic inflammation. Our in vitro system proved to reproduce inflammatory events observed in vivo, such as vasodilation, increased number of macrophages and mast cells, and increased cytokine secretion. This bioreactor-based system may therefore be suitably and reliably used to simulate in vitro human skin inflammation and may be foreseen as a promising tool to test the efficacy of drugs and cosmetics.

Background: Obesity is a pandemic disease characterized by excessive severe body comorbidities. Reduction in fat accumulation represents a mechanism of prevention, and the replacement of white adipose tissue (WAT) with brown adipose tissue (BAT) has been proposed as one promising strategy against obesity. In the present study, we sought to investigate the ability of a natural mixture of polyphenols and micronutrients (A5+) to counteract white adipogenesis by promoting WAT browning. Methods: For this study, we employed a murine 3T3-L1 fibroblast cell line treated with A5+, or DMSO as control, during the differentiation in mature adipocytes for 10 days. Cell cycle analysis was performed using propidium iodide staining and cytofluorimetric analysis. Intracellular lipid contents were detected by Oil Red O staining. Inflammation Array, along with qRT-PCR and Western Blot analyses, served to measure the expression of the analyzed markers, such as pro-inflammatory cytokines. Results: A5+ administration significantly reduced lipids’ accumulation in adipocytes when compared to control cells (p < 0.005). Similarly, A5+ inhibited cellular proliferation during the mitotic clonal expansion (MCE), the most relevant stage in adipocytes differentiation (p < 0.0001). We also found that A5+ significantly reduced the release of pro-inflammatory cytokines, such as IL-6 and Leptin (p < 0.005), and promoted fat browning and fatty acid oxidation through increasing expression levels of genes related to BAT, such as UCP1 (p < 0.05). This thermogenic process is mediated via AMPK-ATGL pathway activation. Conclusion: Overall, these results demonstrated that the synergistic effect of compounds contained in A5+ may be able to counteract adipogenesis and then obesity by inducing fat browning.

Bladder cancer is very common in humans and is often characterized by recurrences, compromising the patient’s quality of life with a substantial social and economic impact. Both the diagnosis and treatment of bladder cancer are problematic due to the exceptionally impermeable barrier formed by the urothelium lining the bladder; this hinders the penetration of molecules via intravesical instillation while making it difficult to precisely label the tumor tissue for surgical resection or pharmacologic treatment. Nanotechnology has been envisaged as an opportunity to improve both the diagnostic and therapeutic approaches for bladder cancer since the nanoconstructs can cross the urothelial barrier and may be functionalized for active targeting, loaded with therapeutic agents, and visualized by different imaging techniques. In this article, we offer a selection of recent experimental applications of nanoparticle-based imaging techniques, with the aim of providing an easy and rapid technical guide for the development of nanoconstructs to specifically detect bladder cancer cells. Most of these applications are based on the well-established fluorescence imaging and magnetic resonance imaging currently used in the medical field and gave positive results on bladder cancer models in vivo, thus opening promising perspectives for the translation of preclinical results to the clinical practice.

The 15th edition of the international conference ArcheoFOSS Open Software, hardware, processes, data, and formats in archaeology is now open. After the 2020’s conference is the second year in which the official conference languages are both English and Italian. The conference will address a range of important topics and themes relating to data sharing, open tools, processes and formats in the archaeological domain. The Organizing Committee invited scholars, independent researchers, institutions, freelance archaeologists, and professional companies involved in Cultural Heritage to submit their original researches or case studies by exposing the latest trends, theoretical or practical developments and challenges in the field. On behalf of the Scientific Committee 2021, the Organising Committee encouraged abstract proposals focused on some thematic areas. Furthermore, for the first time, ArcheoFOSS published the pre-acs volume to facilitate the sharing and the deep understanding of any issues.

The investigation of the absorption of drug delivery systems, designed for the transport of therapeutic molecules inside the body, could be relatively simplified by the fluorophore association and tracking by means of bio-imaging techniques (i.e., optical in vivo imaging or confocal and multiphoton microscopy). However, when a fluorescence signal comes out from the skin, its specific detection can be problematic. Skin high autofluorescence can hinder the observation of administered exogenous fluorophores conjugated to drug delivery systems, making it more challenging to detect their biodistribution. In the present study, we have developed a method based on the spectrofluorometric analysis of skin samples to discriminate the fluorescent signal coming from administered fluorescent molecules from the background. Moreover, we gave a semi-quantitative evaluation of the signal intensity. Thus, we distinguished two gel formulations loading the fluorophore rhodamine B (called GEL RHO and GEL SLN-RHO). The two formulations of gels, one of which containing solid lipid nanoparticles (GEL RHO-SLN), were administered on skin explants incubated in a bioreactor, and the penetration was evaluated at different time points (2 and 6 hours). Cryostatic sections of skin samples were observed with confocal laser scanning microscopy, and a spectrofluorometric analysis was performed. Significantly higher signal intensity in the samples administered with SLN-RHO GEL, with a preferential accumulation in the hair bulbs, was found. Reaching also the deeper layers of the hair shaft after 6 hours, the solid lipid nanoparticles thickened with polymer represent a suitable drug delivery system for transcutaneous administration.

Abstract Background Few cases of scedosporiosis have been reported in animals, but the true prevalence is probably underestimated due to a lack of awareness. Scedosporiosis in dogs has often been associated with localized infection (i.e., nasal infection, eumycetoma, or keratomycosis) or, in rare cases, disseminated infections. Case presentation This case report describes the clinical and pathological features and the diagnostic process of a rare systemic and fatal fungal infection in a dog caused by Scedosporium apiospermum. A 10-month-old female Maremmano-Abruzzese sheepdog showing weakness, lethargy, lateral decubitus, miosis and muscular rigidity was presented. Rodenticide poisoning was clinically suspected for the differential diagnosis. However, postmortem examinations revealed the presence of a swollen and soft subcutaneous nodule located near the right inguinal breast, which was associated with massive enlargement of the inguinal lymph nodes and small disseminated, cream-colored nodules in the kidneys and mesentery. Multiple fungal pyogranulomas were observed upon histological examination. Fungal isolation from the kidneys, breast and inguinal lymph nodes was performed. The internal transcribed spacer (ITS) sequences from the fungal colony DNA were searched in BLAST in the NCBI GenBank for species identification. The sequences of the fungi isolated from the kidney and breast cultures showed 100% sequence identity with sequences from Scedosporium apiospermum. Conclusions This report shows that Scedosporium apiospermum may act as a primary pathogen in young and apparently healthy dogs and represents an important pathogen that should be considered during the diagnostic process, particularly when a fungal infection is suspected.

Abstract Background Elevated cholesterol levels and systemic inflammation are considered relevant risk factors for cardiovascular disease (CVD) development and progression. Increasing evidence suggests that cholesterol-lowering and inflammation-lowering nutraceuticals are useful in the management of moderate hypercholesterolemia. Here, we describe the study protocol of a clinical trial aimed to evaluate the cholesterol and inflammatory lowering effect of an innovative dietary supplement (BruMeChol™, Mivell S.r.l., Italy), composed of a mixture of extracts of bergamot and olive fruits in association with vitamin K2 in subjects with mild hypercholesterolemia. Methods The study was planned as a randomized, double-blind, placebo-controlled, parallel group clinical trial for 12 weeks at the Cardiology Unit of the IRCCS INRCA of Ancona, Italy. A total of 125 subjects (age ≥ 40 years) with mild hypercholesterolemia (total serum cholesterol levels ≥ 200 and ≤ 250 mg/dl) will be recruited. Intervention arm participants will take one capsule of dietary supplement two times a day, 15 min before the main meal. Control arm participants will receive one capsule of placebo in the same way. The dietary supplement capsule contains the following ingredients: phytosterols, flavonoid-rich extract of bergamot fruit (Citrus bergamia), flavonoid-rich extract of olive fruit (Olea europaea), and vitamin K2. Participants will undergo a medical evaluation and chemical-clinical examinations, which include lipid profile, glycemia, biomarkers of renal, liver and cardiac/muscular functions, interleukins (IL 6, IL-32, IL-37, and IL-38), and innovative mediators of inflammation such as inflamma-miRs (miR-21 and miR-146a), at baseline, and after 6 and 12 weeks of treatment. The decrease in total cholesterol levels and inflammatory biomarkers will be the primary and secondary endpoints of the study. Discussion This protocol study, planned to verify the effects of BruMeChol™ dietary supplementation in subjects with mild hypercholesterolemia, could also contribute to new study designs for next large-scale multicenter clinical trials. Trial registration Australian New Zealand Clinical Trials Registry: ACTRN12619000170123 . Retrospectively registered on 5 February 2019

Abstract Three relatives carrying a t(4;8)(p15.2;p23.2) translocation had juvenile myoclonic epilepsy, self‐limited photosensitive occipital epilepsy and migraine with aura. The t(4;8) translocation interrupted the coding sequence of CSMD1 gene and occurred immediately to the 3’UTR of STIM2 gene. STIM2 was overexpressed in the patient carrying the unbalanced translocation, and all three individuals had a single functional copy of CSMD1. Array CGH study disclosed that these three individuals also carried a deletion at 5q12.3 that involves the RGS7BP gene. The overall results favor the view that CSMD1, STIM2, and RGS7BP genes could contribute to epilepsy and migraine phenotypes in our family.