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Abstract This ex vivo study devised an analytical ex vivo method for infection/disinfection of simulated lateral canals located in the middle and apical segments of the root. The antibacterial effects of supplementary approaches were tested in this model. Extracted mandibular premolars had their main root canals enlarged and then two lateral canals (100 μm in diameter) were created in the root, one in the apical and the other in the middle portion. Micro-computed tomography was used for specimen selection and to confirm the quality of the simulated ramifications. The specimens were contaminated with a mixed bacterial culture from subgingival bacterial biofilm added to pure Enterococcus faecalis strain ATCC 29212 grown overnight, using special strategies to facilitate culture medium penetration within the lateral canals. The following procedures were tested for disinfection: NaOCl/passive ultrasonic irrigation (PUI), NaOCl/XP-endo Finisher, ozonated water/continuous ultrasonic irrigation (CUI), and NaOCl/conventional irrigation with 30-G needles (control). Bacteriological samples were taken from the main canal before (S1) and after (S2) each supplementary protocol, and also from each lateral canal after treatment (S3). DNA extracted from the samples was subjected to quantitative real-time polymerase chain reaction. All S1 main canal samples were positive for bacterial presence. Bacterial counts in the main root canal substantially decreased by 99.2% after PUI, 99.1% after ozone/CUI, 99% after XP-endo Finisher, and 96% in the control group (P 0.05). The same was observed when comparing the effects of the supplementary approaches in the apical and middle lateral canals (P > 0.05). Only a few lateral canals showed no detectable bacteria. The method proposed here proved effective for ex vivo infection/disinfection studies. All supplementary approaches induced a substantial bacterial reduction in the main canal, with no significant differences between them. However, in terms of lateral canal disinfection, none of the tested approaches showed significant effects when compared to the control group.

Paclobutrazol (PBZ) is a growth regulator that widely used in horticulture and in the tomato seedling growth to compact the shoots, increase the stem diameter and, root biomass, allowing more tolerance of the seedlings against adverse weather conditions. The objective of this work was to evaluate the rates of paclobutrazol (0, 4, 7, 10 and 13 mg L-1) applied 15 days after sowing by foliar spray on the growth, chemical composition and xylem vessel number of tomato seedlings cultivated in two periods. The PBZ regardless of the application rate reduced the height of tomato seedlings in both growth periods. The basal stem diameter and leaf area were increased with 13 mg L-1 of PBZ. The lignin percentage also increased with 10 and 13 mg L-1 of PBZ as compared to control for both periods. The number of xylem vessel was not affected by PBZ application on the seedlings in the first period. PBZ application at rates of 7 and 10 mg L-1 increased the xylem vessel number in the second period. In general, the application of 13 mg L-1 of PBZ generated seedling more robust to overcome climate adversities.

ABSTRACT The limited temporal completeness and taxonomic accuracy of species lists, made available in a traditional manner in scientific publications, has always represented a problem. These lists are invariably limited to a few taxonomic groups and do not represent up-to-date knowledge of all species and classifications. In this context, the Brazilian megadiverse fauna is no exception, and the Catálogo Taxonômico da Fauna do Brasil (CTFB) (http://fauna.jbrj.gov.br/), made public in 2015, represents a database on biodiversity anchored on a list of valid and expertly recognized scientific names of animals in Brazil. The CTFB is updated in near real time by a team of more than 800 specialists. By January 1, 2024, the CTFB compiled 133,691 nominal species, with 125,138 that were considered valid. Most of the valid species were arthropods (82.3%, with more than 102,000 species) and chordates (7.69%, with over 11,000 species). These taxa were followed by a cluster composed of Mollusca (3,567 species), Platyhelminthes (2,292 species), Annelida (1,833 species), and Nematoda (1,447 species). All remaining groups had less than 1,000 species reported in Brazil, with Cnidaria (831 species), Porifera (628 species), Rotifera (606 species), and Bryozoa (520 species) representing those with more than 500 species. Analysis of the CTFB database can facilitate and direct efforts towards the discovery of new species in Brazil, but it is also fundamental in providing the best available list of valid nominal species to users, including those in science, health, conservation efforts, and any initiative involving animals. The importance of the CTFB is evidenced by the elevated number of citations in the scientific literature in diverse areas of biology, law, anthropology, education, forensic science, and veterinary science, among others.

Background/aim The participation of individuals with physical impairment in sports has numerous benefits, yet there is also the risk of sustaining sport-related injuries or illnesses. Therefore, prevention programmes of these problems are needed to ensure that individuals can maintain a healthy, active lifestyle. Currently, very few prevention interventions are accessible for these athletes. Therefore, the article aims to describe the development process of the Tailored Injury Prevention in Adapted Sports intervention, an online tailored injury and illness prevention intervention for athletes with a physical impairment.Methods The development was guided by the Knowledge Transfer Scheme (KTS).Results In the first step, a cohort study and a qualitative study were conducted to define the problem statement. In the second step, a systematic review was performed in order to learn from theory. Steps 3 and 4 involved an iterative process involving collaboration with diverse expert groups. This included defining athletes’ needs and creating a health problem blueprint, after which the intervention content was created. To ensure accuracy and completeness, a feedback loop was incorporated. In the final phase of this step, we refined the language used within the intervention together with athletes. Finally, an effect and process evaluation will take place in the last step of the KTS.Conclusions Through a five-step approach of the KTS, we developed an online injury and illness prevention intervention for athletes with a physical impairment. This intervention provides direct, timely feedback based on their current health status. Furthermore, it takes the sport and the physical impairment of the athletes into account with regard to the given prevention advices.

Abstract Background Multimorbidity is increasing among adults in the United States. Yet limited research has examined multimorbidity clusters in persons aged 50 years and older with and without a history of cancer. An increased understanding of multimorbidity clusters may improve the cancer survivorship experience for survivors with multimorbidity. Methods We identified 7580 adults aged 50 years and older with 2 or more diseases—including 811 adults with a history of primary breast, colorectal, cervical, prostate, or lung cancer—from the 2018 National Health Interview Survey. Exploratory factor analysis identified clusters of multimorbidity among cancer survivors and individuals without a history of cancer (controls). Frequency tables and chi-square tests were performed to determine overall differences in sociodemographic characteristics, health-related characteristics, and multimorbidity between groups. Results Cancer survivors reported a higher prevalence of having 4 or more diseases compared to controls (57% and 38%, respectively). Our analysis identified 6 clusters for cancer survivors and 4 clusters for controls. Three clusters (pulmonary, cardiac, and liver) included the same diseases for cancer survivors and controls. Conclusions Diseases clustered differently across adults ≥ 50 years of age with and without a history of cancer. Findings from this study may be used to inform clinical care, increase the development and dissemination of multilevel public health interventions, escalate system improvements, and initiate innovative policy reform.

BackgroundNegative body image and adverse body self-evaluation represent key psychological constructs within the realm of weight bias (WB), potentially intertwined with the negative self-evaluation characteristic of depressive symptomatology. Although WB encapsulates an implicit form of self-critical assessment, its exploration among people with mood disorders (MD) has been under-investigated. Our primary goal is to comprehensively assess both explicit and implicit WB, seeking to reveal specific dimensions that could interconnect with the symptoms of MDs.MethodsA cohort comprising 25 MD patients and 35 demographically matched healthy peers (with 83% female representation) participated in a series of tasks designed to evaluate the congruence between various computer-generated body representations and a spectrum of descriptive adjectives. Our analysis delved into multiple facets of body image evaluation, scrutinizing the associations between different body sizes and emotionally charged adjectives (e.g., active, apple-shaped, attractive).ResultsNo discernible differences emerged concerning body dissatisfaction or the correspondence of different body sizes with varying adjectives. Interestingly, MD patients exhibited a markedly higher tendency to overestimate their body weight (p = 0.011). Explicit WB did not show significant variance between the two groups, but MD participants demonstrated a notable implicit WB within a specific weight rating task for BMI between 18.5 and 25 kg/m2 (p = 0.012).ConclusionsDespite the striking similarities in the assessment of participants’ body weight, our investigation revealed an implicit WB among individuals grappling with MD. This bias potentially assumes a role in fostering self-directed negative evaluations, shedding light on a previously unexplored facet of the interplay between WB and mood disorders.

BackgroundLupus pathogenesis is mainly ascribed to increased production and/or impaired clearance of dead cell debris. Although self-reactive T and B lymphocytes are critically linked to lupus development, neutrophils, monocytes, and natural killer (NK) cells have also been implicated. This study assessed apoptosis-related protein expressions in NK cells of patients with juvenile-onset systemic lupus erythematosus (jSLE) and relations to disease activity parameters, nephritis, and neuropsychiatric involvement.MethodsThirty-six patients with jSLE, 13 juvenile dermatomyositis (JDM) inflammatory controls, and nine healthy controls had Fas, FasL, TRAIL, TNFR1, Bcl-2, Bax, Bim, and caspase-3 expressions in NK cells (CD3−CD16+CD56+) simultaneously determined by flow cytometry. Disease activity parameters included Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) score, erythrocyte sedimentation rate, C-reactive protein level, anti-double strain DNA antibody level, complement fractions C3 and C4 levels.ResultsPatients with jSLE had a profile of significantly reduced expression of TRAIL, Bcl-2, and TNFR1 proteins in NK cells when compared to healthy controls. Similar profile was observed in patients with jSLE with active disease, positive anti-dsDNA, nephritis, and without neuropsychiatric involvement. Patients with jSLE with positive anti-dsDNA also had reduced expression of Bax in NK cells when compared healthy controls and to those with negative anti-dsDNA. Yet, patients with jSLE with negative anti-dsDNA had reduced mean fluorescence intensity (MFI) of Bim in NK cells compared to healthy controls. Patients with jSLE with nephritis also had reduced MFI of Fas in NK cells when compared to those without nephritis. In addition, in patients with jSLE, the proportion of FasL-expressing NK cells directly correlated with the SLEDAI-2K score (rs = 0.6, p = 0.002) and inversely correlated with the C3 levels (rs = −0.5, p = 0.007). Moreover, patients with jSLE had increased NK cell percentage and caspase-3 protein expression in NK cells when compared to JDM controls.ConclusionThis study extends to NK cells an altered profile of TRAIL, Bcl-2, TNFR1, Fas, FasL, Bax, Bim, and caspase-3 proteins in patients with jSLE, particularly in those with active disease, positive anti-dsDNA, nephritis, and without neuropsychiatric involvement. This change in apoptosis-related protein expressions may contribute to the defective functions of NK cells and, consequently, to lupus development. The full clarification of the role of NK cells in jSLE pathogenesis may pave the way for new therapies like those of NK cell–based.

BackgroundThe COVID-19 pandemic had serious impact on the well-being of health care workers and highlighted the need for resources to help hospital staff to cope with psychologically negative consequences. The purpose of this study was to investigate the potentially protective effect of spirituality, as measured by the construct of transpersonal trust, against anxiety in physicians and in hospital pastoral care workers. In addition, transpersonal trust was compared to the effects of other potential resources, namely sense of coherence, optimism, and resilience. We also explored the relationship between transpersonal trust and anxiety and how it was moderated by sense of coherence and expected a significant effect.MethodThe sample included N = 405 participants (n = 151 pastoral care workers and n = 254 physicians) who completed an online survey during the first wave of the COVID-19 pandemic between 20th April and 05th July, 2020, that comprised established questionnaires assessing anxiety, transpersonal trust, sense of coherence, and resilience.ResultsThere was no statistically significant negative relationship between transpersonal trust and anxiety in either profession or broken down by occupational group. Multiple regression analysis revealed that sense of coherence inversely predicted generalized anxiety, while transpersonal trust, resilience, and optimism did not. As hypothesized, the association between transpersonal trust and anxiety was moderated by sense of coherence. However, we could not confirm our hypothesis of a protective effect of transpersonal trust against anxiety.ConclusionOur results point to the significant role of sense of coherence as a protective factor against anxiety and highlight the complexity of the relationship among spirituality, transpersonal trust, and anxiety.

BackgroundThe COVID-19 pandemic presents a significant challenge to professional responders in healthcare settings. This is reflected in the language used to describe the pandemic in the professional literature of the respective professions. The aim of this multidisciplinary study was to analyze the linguistic imagery in the relevant professional literature and to determine the identification of different professional groups with it and its emotional effects.MethodA list of 14 typical, widespread and differing imageries for COVID-19 in form of single sentences (e.g., “Until the pandemic is over, we can only run on sight.”) were presented to 1,795 healthcare professionals in an online survey. The imageries had been extracted from a qualitative search in more than 3,500 international professional journals in medicine, psychology and theology. Ratings of agreement with these imageries and feelings about them were subjected to factor analysis.ResultsBased on the list of imageries presented, it was possible to identify three factors for high/low agreement by experiences, and two factors for high/low induced feelings. Broad agreement emerged for imageries on “fight against the crisis” and “lessons learned from the crisis”, while imageries on “acceptance of uncontrollability” tended to be rejected. Imageries of “challenges” tended to lead to a sense of empowerment among subjects, while imageries of “humility” tended to lead to a sense of helplessness.ConclusionBased on the qualitative and subsequential quantitative analysis, several factors for imageries for the COVID-19 pandemic were identified that have been used in the literature. Agreement with imageries is mixed, as is the assessment of how helpful they are.

Abstract Background Essential elements have functions in tumor progression by promoting protumoral cellular processes, such as proliferation, and migration, among others. Obtaining an understanding of how these elements relate to tumor progression processes is of great importance for research. Elemental profile studies in distant tissues, which can be modulated by tumor cells to promote metastasis, have not been sufficiently investigated. The main goal of this study is to evaluate multielemental distribution during tumor progression, focusing on tumor tissue and distant tissues that may be affected. Methods Tumor progression in vivo was simulated by inoculating C57BL/6 mice with Lewis Lung Carcinoma (LLC) cells. Samples of the primary tumor and distant tissues were collected during 5 weeks of tumor progression for the control and experimental (tumor-bearing) groups. The biological samples were analyzed using the synchrotron radiation X-Ray fluorescence technique. Data on the concentration of P, S, K, Ca, Mn, Fe, Cu, and Zn in the samples were obtained and statistically analyzed to evaluate the distribution of the elements during tumor progression in the primary tumor as well as distant tissues. Results It was possible to observe significant changes in the concentrations’ distribution of P, S, K, Ca, Mn, Fe, and Cu in distant tissues caused by the presence of tumor cells. It was also possible to detect a greater similarity between tumor tissue (which has the lung as tissue of origin) and a tissue of non-origin, such as the liver, which is an unprecedented result. Moreover, changes in the distributions of concentrations were detected and studied over time for the different tissues analyzed, such as primary tumor, liver and lung, in Control and Tumor groups. Conclusions Among other results, this paper could explore the modulation of distant tissues caused by the presence of a primary tumor. This could be achieved by the evaluation of several elements of known biological importance allowing the study of different biological processes involved in cancer. The role of essential elements as modulators of the tumor microenvironment is a relevant aspect of tumor progression and this work is a contribution to the field of tumoral metallomics.

OBJECTIVES/GOALS: UNC-Chapel Hill’s CTSA implemented a community feedback session (CFS) service to help researchers gather actionable, authentic, and constructive feedback on their projects from community experts. Simultaneously, we conducted an ongoing, participatory evaluation process to improve the experience for researchers and community members. METHODS/STUDY POPULATION: Informed by the Community Engagement Studio model (Yoosten, 2015), a CFS is a 2-hour consultative session that includes facilitated discussion with community experts around topics or questions posed by a research team. UNC-Chapel Hill’s CTSA staff conducted 7 CFSs during the pilot phase of the service and collected evaluation surveys from researchers and community experts. We held a data party – a participatory evaluation method – to analyze and interpret survey data. Resulting recommendations were used to improve CFS materials and processes. We conducted 11 CFSs after the pilot phase, then analyzed survey data again to gauge improvement and make further adjustments to the service. We also surveyed researchers three months after their last CFS to assess the impact of community experts’ feedback on their studies. RESULTS/ANTICIPATED RESULTS: Since January 2022, 108 community experts have participated in 18 CFSs spanning 9 research projects. Data party interpretations of pilot evaluation data yielded several changes in service delivery; since those changes were instituted, all researchers have highly rated the service, unanimously recommending it to colleagues. Researchers praised well-structured sessions and productive engagement, citing direct benefits to their work and significant impact on recruitment processes 3 months post CFS. Community experts also echoed satisfaction, with 100% finding CFSs worthwhile and 95% desiring to participate again, emphasizing the sessions' supportive atmosphere. A community expert reported that the sessions 'made me feel seen and allowed for a greater understanding of what I have been dealing with.' DISCUSSION/SIGNIFICANCE: The CFS model allows research teams to elicit rapid and meaningful community input, which is key to improving research relevance and impact. Ongoing participatory evaluation of the service ensures continuous improvement, yielding more meaningful interactions and studies that reflect the perspectives of people affected by the research.

Highlights ∙ CTX related to chemotherapy is an important cause of heart failure in surviving cancer patients; ∙ Cardiomyocytes may increase their glycolytic metabolism under significant stress. This phenomenon has been observed in both experimental and clinical studies; ∙ PET/CT is a powerful noninvasive diagnostic technique that provides both anatomical and metabolic information; ∙ 18F-FDG is a glucose analog that is the most commonly used radiopharmaceutical in oncological PET/CT exams; ∙ The degree of myocardial 18F-FDG uptake may be a promising marker of the cardiomyocyte changes that precede cardiac dysfunction in the CTX cascade.

Abstract Seizure Related 6 Homolog Like 2 (SEZ6L2) protein has been shown to have implications in neuronal and especially motor function development. In oncology, overexpression of SEZ6L2 serves as a negative prognostic marker in several tumor entities. Recently, few cases of anti-SEZ6L2 antibody mediated cerebellar syndromes were reported. In this article, we present a case of a 70-year-old woman with subacute onset of gait disturbance, dysarthria and limb ataxia. Serum anti-SEZ6L2 antibodies were markedly increased, and further diagnostic workup revealed left sided breast cancer. Neurological symptoms and SEZ6L2 titer significantly improved after curative tumor therapy. This is a very rare and educationally important report of anti-SEZ6L2 autoimmune cerebellar syndrome with a paraneoplastic etiology. Additionally, we performed a review of the current literature for SEZ6L2, focusing on comparing the published cases on autoimmune cerebellar syndrome.

Next to acute sickness behavior, septic encephalopathy is the most frequent involvement of the brain during infection. It is characterized by a cross-talk of pro-inflammatory cells across the blood–brain barrier, by microglial activation and leukocyte migration, but not by the entry of infecting organisms into the brain tissue. Septic encephalopathy is very frequent in older persons because of their limited cognitive reserve. The predominant clinical manifestation is delirium, whereas focal neurological signs and symptoms are absent. Electroencephalography is a very sensitive method to detect functional abnormalities, but these abnormalities are not specific for septic encephalopathy and of limited prognostic value. Routine cerebral imaging by computer tomography usually fails to visualize the subtle abnormalities produced by septic involvement of the brain. Magnetic resonance imaging is by far more sensitive to detect vasogenic edema, diffuse axonal injury or small ischemic lesions. Routine laboratory parameters most suitable to monitor sepsis, but not specific for septic encephalopathy, are C-reactive protein and procalcitonin. The additional measurement of interleukin (IL)-6, IL-8, IL-10 and tumor necrosis factor-α increases the accuracy to predict delirium and an unfavorable outcome. The most promising laboratory parameters to quantify neuronal and axonal injury caused by septic encephalopathy are neurofilament light chains (NfL) and S100B protein. Neuron-specific enolase (NSE) plasma concentrations are strongly influenced by hemolysis. We propose to determine NSE only in non-hemolytic plasma or serum samples for the estimation of outcome in septic encephalopathy.

Different studies corroborate a role for ceramide synthases and their downstream products, ceramides, in modulation of apoptosis and autophagy in the context of cancer. These mechanisms of regulation, however, appear to be context dependent in terms of ceramides’ fatty acid chain length, subcellular localization, and the presence or absence of their downstream targets. Our current understanding of the role of ceramide synthases and ceramides in regulation of apoptosis and autophagy could be harnessed to pioneer the development of new treatments to activate or inhibit a single type of ceramide synthase, thereby regulating the apoptosis induction or cross talk of apoptosis and autophagy in cancer cells. Moreover, the apoptotic function of ceramide suggests that ceramide analogues can pave the way for the development of novel cancer treatments. Therefore, in the current review paper we discuss the impact of ceramide synthases and ceramides in regulation of apoptosis and autophagy in context of different types of cancers. We also briefly introduce the latest information on ceramide synthase inhibitors, their application in diseases including cancer therapy, and discuss approaches for drug discovery in the field of ceramide synthase inhibitors. We finally discussed strategies for developing strategies to use lipids and ceramides analysis in biological fluids for developing early biomarkers for cancer.

This study aimed to characterize 300 Aeromonas spp. strains isolated from 123 ornamental fish of 32 different species presenting with septicemia, skin lesions, and/or eye lesions. Within the 300 strains, 53.0% were identified as A. veronii, 41.3% as A. hydrophila, and 5.7% as A. caviae. Among the six virulence genes investigated, the most frequent were act (90.3%) and aer (79.3%). More than 50% of A. hydrophila strains were positive for all the studied genes. A total of 30 virulence profiles were identified, with the five main profiles identified comprising 75% of strains. Only five strains were negative for all genes and were identified as A. caviae and A. veronii. The antimicrobial susceptibility profile was performed for 234 strains, with sulfonamides presenting more than 50% of the resistance rates. Susceptibility was observed mainly for cephalosporins, aminoglycosides, chloramphenicol and piperacillin-tazobactam. Multidrug resistance was detected in 82.5% of the studied strains, including A. caviae with 100% multidrug resistance, and A. hydrophila with 90.9% multidrug resistance. The SE-AFLP analysis resulted in 66 genotypes of A. hydrophila, 118 genotypes of A. veronii, and 14 genotypes of A. caviae, demonstrating the greater heterogeneity of A. veronii and A. caviae. However, no direct correlation was observed between the genotypes and the strains’ origins or virulence and resistance profiles.

Astrocytic tumors are known for their high progression capacity and high mortality rates; in this regard, proteins correlated to prognosis can aid medical conduct. Although several genetic changes related to progression from grade 2 to grade 4 astrocytoma are already known, mRNA copies do not necessarily correlate with protein abundance and therefore could shadow further comprehension about this tumor's biology. This motivates us to seek for complementary strategies to study tumor progression at the protein level. Here we compare the proteomic profile of biopsies from patients with grade 2 (diffuse, n = 6) versus grade 4 astrocytomas (glioblastomas, n = 10) using shotgun proteomics. Data analysis performed with PatternLab for proteomics identified 5,206 and 6,004 proteins in the 2- and 4-grade groups, respectively. Our results revealed seventy-four differentially abundant proteins (p < 0.01); we then shortlist those related to greater malignancy. We also describe molecular pathways distinctly activated in the two groups, such as differences in the organization of the extracellular matrix, decisive both in tumor invasiveness and in signaling for cell division, which, together with marked contrasts in energy metabolism, are determining factors in the speed of growth and dissemination of these neoplasms. The degradation pathways of GABA, enriched in the grade 2 group, is consistent with a favorable prognosis. Other functions such as platelet degranulation, apoptosis, and activation of the MAPK pathway were correlated to grade 4 tumors and, consequently, unfavorable prognoses. Our results provide an important survey of molecular pathways involved in glioma pathogenesis for these histopathological groups.

Plain English summary Patients with a severe form of anorexia nervosa (AN) are often treated as inpatients. Many of them benefit from this acute treatment. Unfortunately, a significant number of patients experience relapse after discharge. This problem could be addressed by specific treatments directly following inpatient therapy, so called aftercare interventions, which are tailored to patients’ needs in this treatment period. This review looks at studies which have investigated the efficacy of aftercare interventions for patients with AN directly after inpatient treatment. We included any studies which compared a novel aftercare intervention to a control treatment and where patients were randomly assigned to either of these treatments, as this procedure is considered to reduce bias. We found seven studies that investigated different aftercare intervention approaches, including medication, guided self-help and psychotherapy, and three ongoing studies. Based on the very limited evidence so far, no clear recommendations can be made favoring a specific approach for post-inpatient aftercare in adult patients with AN. The review shows that it should be a priority to increase uptake of aftercare interventions and to reduce dropout rates. This could be achieved by a better tailoring of interventions to patient needs and the integration of patient perspectives in intervention design. More studies are needed to find interventions which allow patients with AN to maintain treatment gains after intensive inpatient treatment.

Abstract Mucopolysaccharidoses (MPS) are rare inborn errors of metabolism, leading to the accumulation of glycosaminoglycans (GAG) in distinct tissues. We investigated oropharyngeal dysphagia using the videofluoroscopic swallowing study (VFSS) in patients with different MPS types. Since there is a lack of studies systematically evaluating this disorder in this population, the use of a standard technique should contribute to better evaluate individuals with MPS. A cross-sectional and observational study enrolling patients followed by an outpatient service for lysosomal diseases at the Genetics Service of the Hospital de Clínicas de Porto Alegre (SGM/HCPA) was conducted. Patients underwent semi-structured interviews, clinical evaluation and VFSS. Nineteen patients were evaluated, including patients with MPS types I (16%), II (42%), IIIb (10%) and IVa (32%). Nearly all patients (95%) presented with oropharyngeal dysphagia in the VFSS. The most frequent findings were impaired chewing during oral phase (94%) and reduced laryngeal elevation in the pharyngeal phase (72%). Oropharyngeal dysphagia constituted a prevalent symptom in the studied cohort regardless of MPS type. Our data reinforces the notion that this disorder should be objectively assessed since it can significantly compromise the nutrition and the hydration of these patients as well as lead to tracheobronchial aspiration, thus resulting in aspiration pneumonia and even death eventually.

O ensino de Física Médica evolui a cada ano no Brasil. A necessidade de métodos didáticos e interativos cresce de maneira proporcional visando uma maior motivação e melhor aprendizagem por parte dos alunos. Nos cursos de graduação em Física Médica no Brasil, a disciplina de Física das Radiações é fundamental para a formação profissional do estudante. O objetivo deste trabalho é utilizar o código de Monte Carlo TOPAS para simular experimentos que possibilitem a discussão de conteúdos abordados na disciplina de Física das Radiações. A ferramenta poderá ser utilizada por docentes ao ministrar essa disciplina e por alunos que queiram aprofundar seus conhecimentos e serem introduzidos ao método de Monte Carlo. Inicialmente, foi realizada a validação do TOPAS para os espectros de fótons utilizados através da comparação com dados da literatura. Com isso, foi possível elaborar os arquivos de entrada e realizar a análise de dados dos arquivos de saída para diferentes configurações de irradiação abordadas academicamente de modo a evidenciar a influência de fatores como: estatística das curvas pelo número de histórias; tamanho do campo de irradiação; meios homogêneos e heterogêneos; feixes incidentes de diferentes naturezas. Os resultados obtidos mostram que as simulações elaboradas possibilitam uma análise correta e dinâmica dos processos de interação da radiação com a matéria abordados em Física das Radiações.

During development, the interconnected generation of various neural cell types within the cerebellar primordium is essential. Over embryonic (E) days E9−E13, Purkinje cells (PCs), and cerebellar nuclei (CN) neurons are among the created primordial neurons. The molecular and cellular mechanisms fundamental for the early cerebellar neurogenesis, migration/differentiation, and connectivity are not clear yet. Autophagy has a vital role in controlling cellular phenotypes, such as epithelial-to-mesenchymal transition (EMT) and endothelial to mesenchymal transition (EndMT). Transforming growth factor-beta 1 (TGF-β1) is the main player in pre-and postnatal development and controlling cellular morphological type via various mechanisms, such as autophagy. Thus, we hypothesized that TGF-β1 may regulate early cerebellar development by modifying the levels of cell adhesion molecules (CAMs) and consequently autophagy pathway in the mouse cerebellar primordium. We demonstrated the stimulation of the canonical TGF-β1 signaling pathway at the point that concurs with the generation of the nuclear transitory zone and PC plate in mice. Furthermore, our data show that the stimulated TGF-β1 signaling pathway progressively and chronologically could upregulate the expression of β-catenin (CTNNB1) and N-cadherin (CDH2) with the most expression at E11 and E12, leading to upregulation of chromodomain helicase DNA binding protein 8 (CDH8) and neural cell adhesion molecule 1 (NCAM1) expression, at E12 and E13. Finally, we demonstrated that the stimulated TGF-β signaling pathway may impede the autophagic flux at E11/E12. Nevertheless, basal autophagy flux happens at earlier developmental phases from E9−E10. Our study determined potential role of the TGF-β signaling and its regulatory impacts on autophagic flux during cerebellar development and cadherin expression, which can facilitate the proliferation, migration/differentiation, and placement of PCs and the CN neurons in their designated areas.

Immersive virtual reality technology (VR) still waits for its wide dissemination in research and psychotherapy of eating and weight disorders. Given the comparably high efforts in producing a VR setup, we outline that the technology’s breakthrough needs tailored exploitation of specific features of VR and user-centered design of setups. In this paper, we introduce VR hardware and review the specific properties of immersive VR versus real-world setups providing examples how they improved existing setups. We then summarize current approaches to make VR a tool for psychotherapy of eating and weight disorders and introduce user-centered design of VR environments as a solution to support their further development. Overall, we argue that exploitation of the specific properties of VR can substantially improve existing approaches for research and therapy of eating and weight disorders. To produce more than pilot setups, iterative development of VR setups within a user-centered design approach is needed.

A ketodiacid, 4,4′-dicarboxylate-dicumyl ketone (3), has been intercalated into a Zn, Al layered double hydroxide (LDH) by a coprecipitation synthesis strategy. The structure and chemical composition of the resultant hybrid material (LDH-KDA3) were characterized by powder X-ray diffraction (PXRD), FT-IR, FT-Raman and solid-state 13C{1H} NMR spectroscopies, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), thermogravimetric analysis (TGA), and elemental analysis (CHN). PXRD showed that the dicarboxylate guest molecules assembled into a monolayer to give a basal spacing of 18.0 Å. TGA revealed that the organic guest starts to decompose at a significantly higher temperature (ca. 330°C) than that determined for the free ketodiacid (ca. 230°C). Photochemical experiments were performed to probe the photoreactivity of the ketoacid in the crystalline state, in solution, and as a guest embedded within the photochemically-inert LDH host. Irradiation of the bulk crystalline ketoacid results in photodecarbonylation and the exclusive formation of the radical-radical combination product. Solution studies employing the standard myoglobin (Mb) assay for quantification of released CO showed that the ketoacid behaved as a photoactivatable CO-releasing molecule for transfer of CO to heme proteins, although the photoreactivity was low. No photoinduced release of CO was found for the LDH system, indicating that molecular confinement enhanced the photo-stability of the hexasubstituted ketone. To better understand the behavior of 3 under irradiation, a more comprehensive study, involving excitation of this compound in DMSO-d6 followed by 1H NMR, UV-Vis and fluorescence spectroscopy, was undertaken and further rationalized with the help of time-dependent density functional theory (TDDFT) electronic quantum calculations. The photophysical study showed the formation of a less emissive compound (or compounds). New signals in the 1H NMR spectra were attributed to photoproducts obtained via Norrish type I α-cleavage decarbonylation and Norrish type II (followed by CH3 migration) pathways. TDDFT calculations predicted that the formation of a keto-enol system (via a CH3 migration step in the type II pathway) was highly favorable and consistent with the observed spectral data.

(1) Background: A reduction in the diffusion capacity of the lung for carbon monoxide is a prevalent longer-term consequence of COVID-19 infection. In patients who have zero or minimal residual radiological abnormalities in the lungs, it has been debated whether the cause was mainly due to a reduced alveolar volume or involved diffuse interstitial or vascular abnormalities. (2) Methods: We performed a cross-sectional study of 45 patients with either zero or minimal residual lesions in the lungs (total volume < 7 cc) at two months to one year post COVID-19 infection. There was considerable variability in the diffusion capacity of the lung for carbon monoxide, with 27% of the patients at less than 80% of the predicted reference. We investigated a set of independent variables that may affect the diffusion capacity of the lung, including demographic, pulmonary physiology and CT (computed tomography)-derived variables of vascular volume, parenchymal density and residual lesion volume. (3) Results: The leading three variables that contributed to the variability in the diffusion capacity of the lung for carbon monoxide were the alveolar volume, determined via pulmonary function tests, the blood vessel volume fraction, determined via CT, and the parenchymal radiodensity, also determined via CT. These factors explained 49% of the variance of the diffusion capacity, with p values of 0.031, 0.005 and 0.018, respectively, after adjusting for confounders. A multiple-regression model combining these three variables fit the measured values of the diffusion capacity, with R = 0.70 and p < 0.001. (4) Conclusions: The results are consistent with the notion that in some post-COVID-19 patients, after their pulmonary lesions resolve, diffuse changes in the vascular and parenchymal structures, in addition to a low alveolar volume, could be contributors to a lingering low diffusion capacity.

Three new species of Urocleidoides Mizelle & Price, 1964 are described from the gills of characiform fishes in the Tocantins River and its tributaries. Urocleidoides boulengerellae sp. nov. is described from Boulengerella cuvieri (Spix & Agassiz, 1829) and differs from all its congeners by the dorsal bar with a long posteromedial projection; male copulatory organ with 2–3 counterclockwise rings and a base with a flange; an accessory piece comprising a robust Y-shaped unit and a sheath-like unit; and a highly sclerotized vaginal canal. Urocleidoides paratriangulus sp. nov., described from Psectrogaster amazonica Eigenmann & Eigenmann, 1889, Cyphocharax gouldingi Vari, 1992, Caenotropus labyrinthicus (Kner, 1858) and Mylesinus paucisquamatus Jégu & Santos, 1988, is most similar to Urocleidoides triangulus (Suriano, 1981) Rossin & Timi, 2016 based on the shape of the anchors and bars but differs from U. triangulus in the morphology of the projection of the dorsal bar, the number of rings of male copulatory organ, and by the smaller size of members of hook pairs 1 and 5 compared with those of the remaining pairs. Urocleidoides tocantinensis sp. nov. is easily distinguished from all other species of the genus by the morphology of the vagina, which present a vaginal vestibule with a membranous cap. Urocleidoides triangulus is reported from its type host in the Guandu River, state of Rio de Janeiro. The present study increases the number of Urocleidoides species to 37 recognized species that fit all the generic characters.

Abstract Circulating cell-free DNA (cfDNA) has been investigated as a screening tool for many diseases. To avoid expensive and time-consuming DNA isolation, direct quantification PCR assays can be established. However, rigorous validation is required to provide reliable data in the clinical and non-clinical context. Considering the International Organization for Standardization, as well as bioanalytical method validation guidelines, we provide a comprehensive procedure to validate assays for cfDNA quantification from blood plasma without DNA isolation. A 90 and 222 bp assay was validated to study the kinetics of cfDNA after exercise in patients with systemic lupus erythematosus (SLE). The assays showed ultra-low limit of quantification (LOQ) with 0.47 and 0.69 ng/ml, repeatability ≤ 11.6% (95% CI 8.1–20.3), and intermediate precision ≤ 12.1% (95% CI 9.2–17.7). Incurred sample reanalysis confirmed the precision of the procedure. The additional consideration of pre-analytical factors shows that centrifugation speed and temperature do not change cfDNA concentrations. In SLE patients cfDNA increases ~ twofold after a walking exercise, normalizing after 60 min of rest. The established assays allow reliable and cost-efficient quantification of cfDNA in minute amounts of plasma in the clinical setting. Additionally, the assay can be used as a tool to determine the impact of pre-analytical factors and validate cfDNA quantity and quality of isolated samples.

Selective serotonin re-uptake inhibitors (SSRIs) are one of the most commonly prescribed classes of antidepressants used for the treatment of moderate to severe depressive disorder, personality disorders and various phobias. This class of antidepressants was created with improved margins of safety. However, genetic polymorphism may be responsible for the high variability in patients’ responses to treatment, ranging from failure to delayed therapeutic responses to severe adverse effects of treatment. It is crucial that the appropriate amount of SSRI drugs is administered to ensure the optimum therapeutic efficacy and intervention to minimise severe and toxic effects in patients, which may be the result of accidental and deliberate cases of poisoning. Determining SSRI concentration in human fluids and the environment with high sensitivity, specificity and reproducibility, and at a low cost and real-time monitoring, is imperative. Electrochemical sensors with advanced functional materials have drawn the attention of researchers as a result of these advantages over conventional techniques. This review article aims to present functional materials such as polymers, carbon nanomaterials, metal nanomaterials as well as composites for surface modification of electrodes for sensitive detection and quantification of SSRIs, including fluoxetine, citalopram, paroxetine, fluvoxamine and sertraline. Sensor fabrication, sensor/analyte interactions, design rationale and properties of functional material and the electrocatalytic effect of the modified electrode on SSRI detection are discussed.

Abstract Background The SARS-Coronavirus-2 (SARS-CoV-2) invades the respiratory system, causing acute and sometimes severe pulmonary symptoms, but turned out to also act multisystematically with substantial impact on the brain. A growing number of studies suggests a diverse spectrum of neurological manifestations. To investigate the spectrum of symptoms, we here describe the neurological manifestations and complications of patients with proven SARS-CoV-2 infection who have been hospitalized at the RWTH University Hospital Aachen, Germany. Methods Between March and September 2020, we evaluated common symptoms, clinical characteristics, laboratory (including cerebrospinal fluid (CSF) analysis), radiological, and electroencephalography (EEG) data from 53 patients admitted with a positive SARS-CoV-2 polymerase chain reaction (PCR). We used the Montreal Cognitive Assessment Test (MoCA) to screen for cognitive impairment, when feasible. We compared critically ill and non-critically ill patients categorized according to the presence of Acute Respiratory Distress Syndrome (ARDS). Results Major clinical neurological features of hospitalized COVID-19 patients were coordination deficits (74%), cognitive impairment (61.5%), paresis (47%), abnormal reflex status (45%), sensory abnormalities (45%), general muscle weakness and pain (32%), hyposmia (26%), and headache (21%). Patients with ARDS were more severely affected than non-ADRS patients. 29.6% of patients with ARDS presented with subarachnoid bleedings, and 11.1% showed ischemic stroke associated with SARS-CoV-2 infection. Cognitive deficits mainly affected executive functions, attention, language, and delayed memory recall. We obtained cerebrospinal fluid (CSF) by lumbar puncture in nine of the 53 patients, none of which had a positive SARS-CoV-2 PCR. Conclusions In line with previous findings, our results provide evidence for a range of SARS-CoV-2-associated neurological manifestations. 26% of patients reported hyposmia, emphasizing the neuro-invasive potential of SARS-CoV-2, which can enter the olfactory bulb. It can therefore be speculated that neurological manifestations may be caused by direct invasion of the virus in the CNS; however, PCR did not reveal positive intrathecal SARS-CoV-2. Therefore, we hypothesize it is more likely that the para-infectious severe pro-inflammatory impact of COVID-19 is responsible for the neurological deficits including cognitive impairment. Future studies with comprehensive longitudinal assessment of neurological deficits are required to determine potential long-term complications of COVID-19.

Resumo Fundamento Ainda não está estabelecido se a captação de fluorodesoxiglicose no miocárdio ocorre exclusivamente por características fisiológicas ou se representa um desarranjo metabólico causado pela quimioterapia. Objetivo Investigar os efeitos da quimioterapia no coração dos pacientes com linfoma por tomografia por emissão de pósitrons associada a tomografia computadorizada (PET/CT) com 2-[18F]-fluoro-2-desoxi-D-glicose (18F-FDG PET/CT) antes, durante e/ou após a quimioterapia. Métodos Setenta pacientes com linfoma submetidos a 18F-FDG PET/CT foram retrospectivamente analisados. O nível de significância foi de 5%. A captação de 18F-FDG foi avaliada por três medidas: captação máxima no ventrículo esquerdo ( standardized uptake value , SUV max), razão SUV cardíaco / aorta e SUV cardíaco / SUV no fígado. Também foram comparados peso corporal, glicemia de jejum, tempo pós-injeção e dose administrada de 18F-FDG entre os exames. Resultados A idade média foi de 50,4 ± 20,1 anos e 50% dos pacientes eram mulheres. A análise foi realizada em dois grupos – PET/CT basal vs. intermediário e PET/CT basal vs pós-terapia. Não houve diferença significativa entre as variáveis clínicas e do protocolo dos exames entre os diferentes momentos avaliados. Nós observamos um aumento na SUV máxima no ventrículo esquerdo de 3,5±1,9 (basal) para 5,6±4,0 (intermediário), p=0,01, e de 4,0±2,2 (basal) para 6,1±4,2 (pós-terapia), p

Abstract Background Bacterial meningitis is still a cause of severe neurological disability. The brain is protected from penetrating pathogens by the blood-brain barrier and the innate immune system. The invading pathogens are recognized by pattern recognition receptors including the G-protein-coupled formyl peptide receptors (FPRs), which are expressed by immune cells of the central nervous system. FPRs show a broad spectrum of ligands, including pro- and anti-inflammatory ones. Here, we investigated the effects of the annexin A1 mimetic peptide Ac2-26 in a mouse model of pneumococcal meningitis. Methods Wildtype (WT) and Fpr1- and Fpr2-deficient mice were intrathecally infected with Streptococcus pneumoniae D39 (type 2). Subsequently, the different mice groups were treated by intraperitoneal injections of Ac2-26 (1 mg/kg body weight) 2, 8, and 24 h post-infection. The extent of inflammation was analyzed in various brain regions by means of immunohistochemistry and real-time reverse transcription polymerase chain reaction (RT-PCR) 30 h post-infection. Results Ac2-26-treated WT mice showed less severe neutrophil infiltration, paralleled by a reduced induction of pro-inflammatory glial cell responses in the hippocampal formation and cortex. While meningitis was ameliorated in Ac2-26-treated Fpr1-deficient mice, this protective effect was not observed in Fpr2-deficient mice. Irrespective of Ac2-26 treatment, inflammation was more severe in Fpr2-deficient compared to Fpr1-deficient mice. Conclusions In summary, this study demonstrates anti-inflammatory properties of Ac2-26 in a model of bacterial meningitis, which are mediated via FPR2, but not FPR1. Ac2-26 and other FPR2 modulators might be promising targets for the development of novel therapies for Streptococcus pneumoniae-induced meningitis.

ABSTRACT: We analyzed 77 Salmonella spp. strains, from which 20 were isolated from broilers (cloacal swabs) and 57 from chickens from slaughterhouses under federal inspection. The following serotypes were identified: Salmonella Saint Paul (29), Salmonella Heidelberg (27), Salmonella Anatum (9), Salmonella Cerro (5), Salmonella Senftenberg (5), Salmonella enterica (O: 4,5) (1) and Salmonella enterica (O: 9.12) (1). Fifteen strains (19.5%) were resistant to enrofloxacin, six (7.8%) to ciprofloxacin, and 26 (33.8%) to nalidixic acid in the Disk Diffusion Test. The fifteen enrofloxacin resistant strains were selected for the PCR to detect the genes gyrA, gyrB, parC, and parE, and genetic sequencing to identify mutations in these genes. Five strains (33.3%) had point mutations in the gyrA gene, and one (6.7%) presented a point mutation in the parC gene. None of the 15 strains had mutations in the gyrB and parE genes, and none had more than one mutation in the gyrA gene or the other genes. The presence of point mutations in the strains studied corroborates with the phenotypic resistance observed to nalidixic acid. However, it did not explain the resistance to fluoroquinolones found in the 15 strains. Other mechanisms may be related to the fluoroquinolones resistance, highlighting the need for additional mutation screening.

Abstract Background An important hallmark of Alzheimer’s disease (AD) is the increase of Aβ1-42 burden and its accumulation to senile plaques, leading the reactive gliosis and neurodegeneration. The modulation of glia cell function represents an attractive therapeutic strategy, but is currently limited by an incomplete understanding of its relevance for AD. The chemotactic G-protein coupled formyl peptide receptor (FPR), which is known to modulate Aβ1-42 uptake and signal transduction, might be one candidate molecule regulating glia function in AD. Here, we investigate whether the modulation of FPR exerts beneficial effects in an AD preclinical model. Methods To address this question, APP/PS1 double-transgenic AD mice were treated for 20 weeks with either the pro-inflammatory FPR agonist fMLF, the FPR1/2 antagonist Boc2 or the anti-inflammatory FPR2 agonist Ac2-26. Spatial learning and memory were evaluated using a Morris water maze test. Immunohistological staining, gene expression studies, and flow cytometry analyses were performed to study neuronal loss, gliosis, and Aß-load in the hippocampus and cortex, respectively. Results FPR antagonism by Boc2-treatment significantly improved spatial memory performance, reduced neuronal pathology, induced the expression of homeostatic growth factors, and ameliorated microglia, but not astrocyte, reactivity. Furthermore, the elevated levels of amyloid plaques in the hippocampus were reduced by Boc2-treatment, presumably by an induction of amyloid degradation. Conclusions We suggest that the modulation of FPR signaling cascades might be considered as a promising therapeutic approach for alleviating the cognitive deficits associated with early AD. Additional studies are now needed to address the downstream effectors as well as the safety profile of Boc2.

Abstract Objectives Recently, defects in the protein kinase mTOR (mammalian target of rapamycin) and its associated pathway have been correlated with hemimegalencephaly (HME). mTOR acts as a central regulator of important physiological cellular functions such as growth and proliferation, metabolism, autophagy, death, and survival. This study was aimed at identifying specific variants in mTOR signaling pathway genes in patients diagnosed with HME. Methods Using amplicon and whole exome sequencing (WES) of resected brain and paired blood samples from five HME patients, we were able to identify pathogenic mosaic variants in the mTOR pathway genes MTOR, PIK3CA, and DEPDC5. Results These results strengthen the hypothesis that somatic variants in PI3K‐Akt‐mTOR pathway genes contribute to HME. We also describe one patient presenting with a pathogenic variant on DEPDC5 gene, which reinforces the role of DEPDC5 on cortical structural changes due to mTORC1 hyperactivation. These findings also provide insights into when in brain development these variants occurred. An early developmental variant is expected to affect a larger number of cells and to result in a larger malformation, whereas the same variant occurring later in development would cause a minor malformation. Significance In the future, numerous somatic variants in known or new genes will undoubtedly be revealed in resected brain samples, making it possible to draw correlations between genotypes and phenotypes and allow for a genetic clinical diagnosis that may help to predict a given patient's outcome.

Coastal systems such as estuaries are threatened by multiple anthropogenic stressors worldwide. However, how these stressors and estuarine hydrology shape benthic bacterial communities and their functions remains poorly known. Here, we surveyed sediment bacterial communities in poorly flushed embayments and well flushed channels in Sydney Harbour, Australia, using 16S rRNA gene sequencing. Sediment samples were collected monthly during the Austral summer-autumn 2014 at increasing distance from a large storm drain in each channel and embayment. Bacterial communities differed significantly between sites that varied in proximity to storm drains, with a gradient of change apparent for sites within embayments. We explored this pattern for embayment sites with analysis of RNA-Seq gene expression patterns and found higher expression of multiple genes involved in bacterial stress response far from storm drains, suggesting that bacterial communities close to storm drains may be more tolerant of localised anthropogenic stressors. Several bacterial groups also differed close to and far from storm drains, suggesting their potential utility as bioindicators to monitor contaminants in estuarine sediments. Overall, our study provides useful insights into changes in the composition and functioning of benthic bacterial communities as a result of multiple anthropogenic stressors in differing hydrological conditions.

Lymphocytes are the central actors in adaptive immune responses. When challenged with antigen, a small number of B and T cells have a cognate receptor capable of recognising and responding to the insult. These cells proliferate, building an exponentially growing, differentiating clone army to fight off the threat, before ceasing to divide and dying over a period of weeks, leaving in their wake memory cells that are primed to rapidly respond to any repeated infection. Due to the non-linearity of lymphocyte population dynamics, mathematical models are needed to interrogate data from experimental studies. Due to lack of evidence to the contrary and appealing to arguments based on Occam’s Razor, in these models newly born progeny are typically assumed to behave independently of their predecessors. Recent experimental studies, however, challenge that assumption, making clear that there is substantial inheritance of timed fate changes from each cell by its offspring, calling for a revision to the existing mathematical modelling paradigms used for information extraction. By assessing long-term live-cell imaging of stimulated murine B and T cells in vitro, we distilled the key phenomena of these within-family inheritances and used them to develop a new mathematical model, Cyton2, that encapsulates them. We establish the model’s consistency with these newly observed fine-grained features. Two natural concerns for any model that includes familial correlations would be that it is overparameterised or computationally inefficient in data fitting, but neither is the case for Cyton2. We demonstrate Cyton2’s utility by challenging it with high-throughput flow cytometry data, which confirms the robustness of its parameter estimation as well as its ability to extract biological meaning from complex mixed stimulation experiments. Cyton2, therefore, offers an alternate mathematical model, one that is, more aligned to experimental observation, for drawing inferences on lymphocyte population dynamics.

Progressive multifocal leukoencephalopathy (PML) is a subacute brain infection by the opportunistic John Cunningham (JC) virus. Herein, we describe seven patients with PML, lymphopenia, and sarcoidosis, in three of whom PML was the first manifestation of sarcoidosis. At onset, the clinical picture comprised rapidly progressive spastic hemi- or limb pareses as well as disturbances of vision, speech, and orientation. Cerebral magnetic resonance imaging showed T2-hyperintense, confluent, mainly supratentorial lesions. Four patients developed punctate contrast enhancement as a radiological sign of an immune reconstitution inflammatory syndrome (IRIS), three of them having a fatal course. In the cerebrospinal fluid, the initial JC virus load (8–25,787 copies/ml) did not correlate with interindividual severity; however, virus load corresponded to clinical dynamics. Brain biopsies ( n = 2), performed 2 months after symptom onset, showed spotted demyelination and microglial activation. All patients had lymphopenia in the range of 270–1150/µl. To control JC virus, three patients received a combination of mirtazapine and mefloquine, another two patients additionally took cidofovir. One patient was treated with cidofovir only, and one patient had a combined regimen with mirtazapine, mefloquine, cidofovir, intravenous interleukin 2, and JC capsid vaccination. To treat sarcoidosis, the four previously untreated patients received prednisolone. Three patients had taken immunosuppressants prior to PML onset, which were subsequently stopped as a potential accelerator of opportunistic infections. After 6–54 months of follow up, three patients reached an incomplete recovery, one patient progressed, but survived so far, and two patients died. One further patient was additionally diagnosed with lung cancer, which he died from after 24 months. We conclude that the combination of PML and sarcoidosis is a diagnostic and therapeutic challenge. PML can occur as the first sign of sarcoidosis without preceding immunosuppressive treatment. The development of IRIS might be an indicator of poor outcome.

Objective: To evaluate the impact of multidimensional interventions on quality of life (QoL) and depressive symptoms in Brazilian older adults living in the community. Methods: Longitudinal, quasi-experimental study of older adults receiving conventional primary health care (PHC). The interventions were designed in response to a first round of data collection and validated through pilot testing in groups of older adults from another community. The validated interventions were then applied to an intervention group (IG). To measure their effect, we used the Medical Outcomes Short-Form Health Survey (SF-36) quality of life scale and the Geriatric Depression Scale (GDS-30). Results: The sample comprised 118 participants. IG participants exhibited significant improvement in several QoL domains (SF-36): mental health (p = 0.010), general health perceptions (p = 0.016), and physical functioning (p = 0.045). No such improvement occurred in controls (p > 0.050). The prevalence of depression (GDS-30) fell from 36.7 to 23.3% in the IG, despite no significant difference (p = 0.272). Controls also reported a reduction in depressive symptoms, but only from 44.8 to 41.4% (p = 0.112). Conclusions: This multidimensional intervention was associated with significant improvement in mental health, general health perceptions, and physical functioning in a sample of Brazilian older adults. Clinical trial registration: RBR-92dbtx.

Abstract Introduction While early diagnostic decision support systems were built around knowledge bases, more recent systems employ machine learning to consume large amounts of health data. We argue curated knowledge bases will remain an important component of future diagnostic decision support systems by providing ground truth and facilitating explainable human-computer interaction, but that prototype development is hampered by the lack of freely available computable knowledge bases. Methods We constructed an open access knowledge base and evaluated its potential in the context of a prototype decision support system. We developed a modified set-covering algorithm to benchmark the performance of our knowledge base compared to existing platforms. Testing was based on case reports from selected literature and medical student preparatory material. Results The knowledge base contains over 2000 ICD-10 coded diseases and 450 RX-Norm coded medications, with over 8000 unique observations encoded as SNOMED or LOINC semantic terms. Using 117 medical cases, we found the accuracy of the knowledge base and test algorithm to be comparable to established diagnostic tools such as Isabel and DXplain. Our prototype, as well as DXplain, showed the correct answer as “best suggestion” in 33% of the cases. While we identified shortcomings during development and evaluation, we found the knowledge base to be a promising platform for decision support systems. Conclusion We built and successfully evaluated an open access knowledge base to facilitate the development of new medical diagnostic assistants. This knowledge base can be expanded and curated by users and serve as a starting point to facilitate new technology development and system improvement in many contexts.

Objective: To validate the Pediatric Obstructive Sleep Apnea Screening tool for use in Brazil. Materials and methods: The Brazilian version of this questionnaire, originally validated and tested in the United States, was developed as follows: (a) translation; (b) back‐translation; (c) completion of the final version; (d) pre‐testing. The questionnaire was applied prior to polysomnography to children aged 3–9 years from October 2015 to October 2016, and its psychometric properties (i.e., validity and reliability) were evaluated. The accuracy was assessed from comparisons between polysomnographic results and corresponding questionnaire scores. Results: Sixty patients were enrolled, and based on polysomnographic findings, 48% patients had normal apnea‐hypopnea index, while the remaining 52% met the criteria for obstructive sleep apnea. Minimum O2 saturation level was obstructive sleep apnea children (p = 0.021). Satisfactory concordance was found between individual apnea-hypopnea index and questionnaire scores. Bland–Altman plot‐derived bias was 0.1 for the difference between measures, with 5.34 (95% CI: 4.14–6.55) and −5.19 (95%CI: −6.39 to −3.98) for the upper and lower agreement range. Internal consistency derived from Cronbach's alpha was 0.84 (95%CI: 0.78–0.90). Conclusion: The questionnaire was translated to and validated into Brazilian‐Portuguese version, and showed good reliability and concordance with apnea‐hypopnea index. This questionnaire offers a reliable screening option for sleep‐disordered breathing in children. Resumo: Objetivo: Validar o questionário Pediatric Obstructive Sleep Apnea Screening Tool para o seu uso no Brasil. Materiais e métodos: A versão brasileira desse questionário, originalmente validado e testado nos Estados Unidos, foi desenvolvida a partir das seguintes etapas: a) tradução; b) retrotradução; c) conclusão da versão final; d) pré‐teste. O questionário foi aplicado previamente ao início da polissonografia em crianças de 3 a 9 anos incluídas no estudo de outubro de 2015 a outubro de 2016. As propriedades psicométricas avaliadas foram validade e confiabilidade. A acurácia foi avaliada pela comparação entre os resultados da polissonografia com o escore do questionário. Resultados: Foram incluídos no estudo 60 pacientes. Conforme a polissonografia, 48% dos pacientes apresentaram índices de apneia e hipopneia normais e 51% apresentaram resultados alterados. A SpO2 mínima foi significativamente menor (p = 0,021) nas crianças com diagnóstico de síndrome de apneia obstrutiva do sono. O índice de apneia e hipopneia apresentou concordância satisfatória com os resultados do questionário. O viés médio de Bland‐Altman foi de 0,1 para a diferença entre as medidas, com um limite superior de 5,34 (IC95%4,14 a 6,55) e um limite inferior de ‐5,19 (IC95%‐6,39 a ‐3,98). A consistência interna do questionário avaliada pelo α de Cronbach foi de 0,84 (IC95%0,78 a 0,90). Conclusão: O questionário foi traduzido e validado adequadamente para a versão em português brasileiro, apresentando boa confiabilidade e concordância com o índice de apneia e hipopneia. Esse questionário oferece uma opção confiável de triagem de distúrbios respiratórios do sono em crianças. Keywords: Questionnaire, Sleep‐disordered breathing, Pediatrics, Validation, Palavras‐chave: Questionário, Distúrbios respiratórios do sono, Pediatria, Validação

Abstract Background Community-based GeneXpert MTB/RIF testing may increase detection of prevalent TB in the community and improve rates of TB treatment completion. Methods We conducted a pilot randomized trial to evaluate the impact of GeneXpert screening on a mobile HIV testing unit. Adults (≥18y) underwent rapid HIV testing and TB symptom screening and were randomized to usual mobile unit care (providing sputum on the mobile unit sent out for GeneXpert testing) or the “Test & Treat TB” intervention with immediate GeneXpert testing. Symptomatic participants in usual care produced sputum that was sent for hospital-based GeneXpert testing; participants were contacted ~ 7 days later with results. In the “Test & Treat TB” intervention, HIV-infected or HIV-uninfected/TB symptomatic participants underwent GeneXpert testing on the mobile unit. GeneXpert+ participants received expedited TB treatment initiation, monthly SMS reminders and non-cash incentives. We assessed 6-month TB treatment outcomes. Results 4815 were eligible and enrolled; median age was 27 years (IQR 22 to 35). TB symptoms included cough (5%), weight loss (4%), night sweats (4%), and fever (3%). 42% of eligible participants produced sputum (intervention: 56%; usual care: 26%). Seven participants tested GeneXpert+, six in the intervention (3%, 95% CI 1%, 5%) and one in usual care (1%, 95% CI 0%, 6%). 5 of 6 intervention participants completed TB treatment; the GeneXpert+ participant in usual care did not. Conclusion GeneXpert MTB/RIF screening on a mobile HIV testing unit is feasible. Yield for GeneXpert+ TB was low, however, the “Test & Treat TB” strategy led to high rates of TB treatment completion. Trial registration This study was registered on November 21, 2014 at ClinicalTrials.gov (NCT02298309).

Objectives: To evaluate the occurrence of drug-related problems (DRP) associates to the use of oral drugs in hospitalized pediatric patients. Method: A cross-sectional study, with data collection performed on the prescriptions of patients using oral medications admitted to the pediatric ward at a reference institute in maternal and child care in Rio de Janeiro. Data collection was carried out from September/2020 to November/2020. Convenience sample was used. Patients aged 0 to 18 years, with oral medications prescribed during the hospitalization period and with a hospital stay of more than 48 hours, were included. DRP were collected and classified based on the pharmacotherapy clinical review guide. Data related to drug interactions were collected in order to identify a possible association with the occurrence of DRP. Pharmaceutical interventions were performed when necessary. Results: Thirty-eight patients were evaluated in this study, most of them male (57.9%) and aged between 0 and 4 years (65.8%). Sixteen patients (42.1%) had at least one DRP. We found 50 DRP, representing a rate of 131.6 DRP per 100 patients. Among the oral medications used, 42.4% were related to DRP. Drugs with action on the nervous system were related to most DRP (37%), with emphasis on the class of antiepileptics. Conclusion: The number of medications used during hospitalization and the number of drug interactions showed statistically significant diferences in patients who had DRP. The review of oral pharmacotherapy identified drug classes and profiles of patients with greater susceptibility to the occurrence of DRP and demonstrated its importance in promoting safety in the use of drugs in the pediatric population.

Peptidoglycan is the major structural component of the Staphylococcus aureus cell wall, in which it maintains cellular integrity, is the interface with the host, and its synthesis is targeted by some of the most crucial antibiotics developed. Despite this importance, and the wealth of data from in vitro studies, we do not understand the structure and dynamics of peptidoglycan during infection. In this study we have developed methods to harvest bacteria from an active infection in order to purify cell walls for biochemical analysis ex vivo. Isolated ex vivo bacterial cells are smaller than those actively growing in vitro, with thickened cell walls and reduced peptidoglycan crosslinking, similar to that of stationary phase cells. These features suggested a role for specific peptidoglycan homeostatic mechanisms in disease. As S. aureus missing penicillin binding protein 4 (PBP4) has reduced peptidoglycan crosslinking in vitro its role during infection was established. Loss of PBP4 resulted in an increased recovery of S. aureus from the livers of infected mice, which coincided with enhanced fitness within murine and human macrophages. Thicker cell walls correlate with reduced activity of peptidoglycan hydrolases. S. aureus has a family of 4 putative glucosaminidases, that are collectively crucial for growth. Loss of the major enzyme SagB, led to attenuation during murine infection and reduced survival in human macrophages. However, loss of the other three enzymes Atl, SagA and ScaH resulted in clustering dependent attenuation, in a zebrafish embryo, but not a murine, model of infection. A combination of pbp4 and sagB deficiencies resulted in a restoration of parental virulence. Our results, demonstrate the importance of appropriate cell wall structure and dynamics during pathogenesis, providing new insight to the mechanisms of disease.

Recent studies have suggested that people's intent and ability to act also can influence their perception of their bodies' peripersonal space. Vice versa one could assume that the inability to reach toward and grasp an object might have an impact on the subject's perception of reaching distance. Here we tested this prediction by investigating body size and action capability perception of neurological patients suffering from arm paresis after stroke, comparing 32 right-brain-damaged patients (13 with left-sided arm paresis without additional spatial neglect, 10 with left-sided arm paresis and additional spatial neglect, 9 patients had neither arm paresis nor neglect) and 27 healthy controls. Nineteen of the group of right hemisphere stroke patients could be re-examined about five months after initial injury. Arm length was estimated in three different methodological approaches: explicit visual, explicit tactile/proprioceptive, and implicit reaching. Results fulfilled the working hypothesis. Patients with an arm paresis indeed perceived their bodies differently. We found a transient overestimation of the length of the contralesional, paretic arm after stroke. Body size and action capability perception for the extremities thus indeed seem to be tightly linked in humans.

State and local health departments have been tasked with promoting the use of face coverings to decrease the spread of COVID-19 in their respective communities. However, little is known about motivations and barriers to wearing face coverings in the context of COVID-19 prevention, particularly among communities of color who are at an increased risk of serious illness from the disease. The purpose of this study was to identify common motivations and barriers to face covering use, as well as explore perceptions of messages encouraging the use of face coverings among a racially and ethnically diverse sample. A survey was distributed electronically to North Carolina (NC) residents through NC Department of Health and Human Services listservs in July 2020. Participants self-categorized as Latino/a (33.5%), Black (39.1%), or white or another race (27.5%). The most commonly endorsed motivations for wearing face coverings were to avoid spreading COVID-19 (77%), as well as to protect people who are vulnerable (76%) and one's community (72%). Being uncomfortable (40%) was the most commonly endorsed barrier. Messages that included a clear request (ex. please wear a face covering) and a direct benefit (ex. keep community safe) were more commonly endorsed than those that did not. Commonly endorsed motivations, behaviors, and messages differed by race and ethnicity. Increased attention to message content, message structure, and access to information and resources may aid local officials in increasing consistent use of face coverings.

Cervical cancer, which main etiologic factor is Human Papillomavirus (HPV) infection, continues to be a burden for public health systems in developing countries. Our laboratory has been working with the hypothesis that signals generated in the tumor microenvironment can modulate local and systemic immune responses. In this context, it would be reasonable to think that tumors create pro-tumoral bias in immune cells, even before they are recruited to the tumor microenvironment. To understand if and how signaling started in the tumor microenvironment can influence cells within the tumor and systemically, we investigated the expression of key proteins in signaling pathways important for cell proliferation, viability, immune responses and tolerance. Besides, we used detection of specific phosphorylated residues, which are indicative of activation for Akt, CREB, p65 NFκB, and STAT3. Our findings included the observation of a significant STAT3 expression increase and p65 NFκB decrease in circulating leukocytes in correlation with lesion grade. In light of those observations, we started investigating the result of the inhibition of STAT3 in a tumor experimental model. STAT3 inhibition impaired tumor growth, increased anti-tumor T cell responses and decreased the accumulation of myeloid cells in the spleen. The concomitant inhibition of NFκB partially reversed these effects. This study indicates that STAT3 and NFκB are involved in immunomodulatory tumor effects and STAT3 inhibition could be considered as therapy for patients with cervical cancer.

Abstract Insulin resistance is a metabolic disorder affecting multiple tissues and is a precursor event to type 2 diabetes (T2D). As T2D affects over 425 million people globally, there is an imperative need for research into insulin resistance to better understand the underlying mechanisms. The proposed mechanisms involved in insulin resistance include both whole body aspects, such as inflammation and metabolic inflexibility; as well as cellular phenomena, such as lipotoxicity, ER stress, and mitochondrial dysfunction. Despite numerous studies emphasizing the role of lipotoxicity in the pathogenesis of insulin resistance, an understanding of the interplay between tissues and these proposed mechanisms is still emerging. Furthermore, the tissue‐specific and unique responses each of the three major insulin target tissues and how each interconnect to regulate the whole body insulin response has become a new priority in metabolic research. With an emphasis on skeletal muscle, this mini‐review highlights key similarities and differences in insulin signaling and resistance between different target‐tissues, and presents the latest findings related to how these tissues communicate to control whole body metabolism.

The home learning environment plays an important role for children’s early competencies development. In particular, the early home literacy environment (HLE) that consists of all literacy resources and interactions in a family that support children’s linguistic and literacy learning is closely associated with children’s language comprehension and production. A key aspect of the HLE is shared reading that should start early in children’s life and should be part of a regular routine in the family. However, parental attitudes toward (shared) reading have hardly been analyzed.In this longitudinal study, we analyzed the associations between parental attitudes toward shared reading and children’s linguistic competencies and whether these associations may be mediated by the HLE. Further, we were interested in changes of parental attitudes over time and their association with child and family background characteristics. The sample consisted of N = 133 children with an average age of about 3 years at t1. Children were tested two more times with a 6-month period in-between each assessment. Parental attitudes toward shared reading, socioeconomic status (SES), and the HLE were assessed via parental survey. Children’s sentence comprehension, productive language, and grammar were measured with a standardized test battery. Children whose parents had a more positive attitude toward shared reading not only lived in a greater quality HLE but also performed better in the linguistic tests. In a structural equation model, an indirect effect was found showing that the HLE mediated the effect of parental attitudes on children’s linguistic competencies. Further, parental attitudes toward shared reading did not change significantly across t1 to t3, and a lower score in the SES scale was associated with a less positive attitude toward shared reading. Consequently, parental attitudes toward shared reading seem to be an important basis for individual differences in the quality of the HLE and also for children’s linguistic competencies. As these attitudes vary in the context of different family SES backgrounds, they may be a good target for interventions to support the quality of the HLE and young children’s linguistic learning.

Background: DuoBody®-CD3xCD20 (GEN3013) is a full-length human IgG1 bispecific antibody (bsAb) recognizing CD3 and CD20, generated by controlled Fab-arm exchange. Its Fc domain was silenced by introduction of mutations L234F L235E D265A. Methods: T-cell activation and T-cell-mediated cytotoxicity were measured by flow cytometry following co-culture with tumour cells. Anti-tumour activity of DuoBody-CD3xCD20 was assessed in humanized mouse models in vivo. Non-clinical safety studies were performed in cynomolgus monkeys. Findings: DuoBody-CD3xCD20 induced highly potent T-cell activation and T-cell-mediated cytotoxicity towards malignant B cells in vitro. Comparison of DuoBody-CD3xCD20 to CD3 bsAb targeting alternative B-cell antigens, or to CD3xCD20 bsAb generated using alternative CD20 Ab, emphasized its exceptional potency. In vitro comparison with other CD3xCD20 bsAb in clinical development showed that DuoBody-CD3xCD20 was significantly more potent than three other bsAb with single CD3 and CD20 binding regions and equally potent as a bsAb with a single CD3 and two CD20 binding regions. DuoBody-CD3xCD20 showed promising anti-tumour activity in vivo, also in the presence of excess levels of a CD20 Ab that competes for binding. In cynomolgus monkeys, DuoBody-CD3xCD20 demonstrated profound and long-lasting B-cell depletion from peripheral blood and lymphoid organs, which was comparable after subcutaneous and intravenous administration. Peak plasma levels of DuoBody-CD3xCD20 were lower and delayed after subcutaneous administration, which was associated with a reduction in plasma cytokine levels compared to intravenous administration, while bioavailability was comparable. Interpretation: Based on these preclinical studies, a clinical trial was initiated to assess the clinical safety of subcutaneous DuoBody-CD3xCD20 in patients with B-cell malignancies. Funding: Genmab Keywords: Bispecific antibody, CD3, CD20, T cell redirection, B cell malignancy, Subcutaneous administration

Rates of teenage pregnancy remain high in sub-Saharan Africa. The KEMRI Health and Demographic Surveillance System provided the sampling frame for a survey. Analysis focused on 1,952 girls aged 13–19 years. Over a third (37.2%; n = 727) were sexually active and 23.3% (n = 454) had ever been pregnant. Adjusted odds of reporting a history of pregnancy were greater for older compared to younger teens, teens who were ever married or cohabiting compared to those who were single, teens with a primary education or less compared to those with a higher level of education, and teens who experienced partner violence in the last 12 months. Three-quarters of teens pregnant in the last 12 months did not want to get pregnant (n = 190); only 64.2% (n = 122) answered yes to using any family planning method. Teen pregnancy and its consequences are serious public health issues. Higher education levels are a crucial component to address the problem.