A non-conserved amino acid variant regulates differential signalling between human and mouse CD28

CD28 superagonistic antibodies (CD28SAb) can preferentially activate and expand immunosuppressive regulatory T cells (Treg) in mice. However, pre-clinical trials assessing CD28SAbs for the therapy of autoimmune diseases reveal severe systemic inflammatory response syndrome in humans, thereby implying the existence of distinct signalling abilities between human and mouse CD28. Here, we show that a single amino acid variant within the C-terminal proline-rich motif of human and mouse CD28 (P212 in human vs. A210 in mouse) regulates CD28-induced NF-κB activation and pro-inflammatory cytokine gene expression. Moreover, this Y209APP212 sequence in humans is crucial for the association of CD28 with the Nck adaptor protein for actin cytoskeleton reorganisation events necessary for CD28 autonomous signalling. This study thus unveils different outcomes between human and mouse CD28 signalling to underscore the importance of species difference when transferring results from preclinical models to the bedside

Stem-like and highly invasive prostate cancer cells expressing CD44v8-10 marker originate from CD44-negative cells

In human prostate cancer (PCa), the neuroendocrine cells, expressing the prostate cancer stem cell (CSC) marker CD44, may be resistant to androgen ablation and promote tumor recurrence. During the study of heterogeneity of the highly aggressive neuroendocrine PCa cell lines PC3 and DU-145, we isolated and expanded in vitro a minor subpopulation of very small cells lacking CD44 (CD44neg). Unexpectedly, these sorted CD44neg cells rapidly and spontaneously converted to a stable CD44high phenotype specifically expressing the CD44v8-10 isoform which the sorted CD44high subpopulation failed to express. Surprisingly and potentially interesting, in these cells expression of CD44v8-10 was found to be induced in stem cell medium. CD44 variant isoforms are known to be more expressed in CSC and metastatic cells than CD44 standard isoform. In agreement, functional analysis of the two sorted and cultured subpopulations has shown that the CD44v8-10pos PC3 cells, resulting from the conversion of the CD44neg subpopulation, were more invasive in vitro and had a higher clonogenic potential than the sorted CD44high cells, in that they produced mainly holoclones, known to be enriched in stem-like cells. Of interest, the CD44v8-10 is more expressed in human PCa biopsies than in normal gland. The discovery of CD44v8-10pos cells with stem-like and invasive features, derived from a minoritarian CD44neg cell population in PCa, alerts on the high plasticity of stem-like markers and urges for prudency on the approaches to targeting the putative CSC

Pain in Multiple System Atrophy a Systematic Review and Meta-Analysis

Background: Individuals with multiple system atrophy (MSA) often complain about pain, nonetheless this remains a poorly investigated non-motor feature of MSA. Objectives: Here, we aimed at assessing the prevalence, characteristics, and risk factors for pain in individuals with MSA. Methods: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes (PRISMA) guidelines, we systematically screened the PubMED, Cochrane, and Web of Science databases for papers published in English until September 30, 2022, combining the following keywords: “pain,” “multiple system atrophy,” “MSA,” “olivopontocerebellar atrophy,” “OPCA,” “striatonigral degeneration,” “SND,” “Shy Drager,” and “atypical parkinsonism.”. Results: The search identified 700 records. Sixteen studies provided information on pain prevalence in cohorts of MSA individuals and were included in a qualitative assessment based on the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) tool. Thirteen studies (11 cross-sectional, two longitudinal) scored ≥14 points on QUADAS assessment and were included in a quantitative analysis, pooling data from 1236 MSA individuals. The resulting pooled prevalence of pain in MSA was 67% (95% confidence intervals [CI] = 57%–75%), and significantly higher in individuals with MSA of parkinsonian rather than cerebellar type (76% [95% CI = 63%–87%] vs. 45% [95% CI = 33%–57%], P = 0.001). Pain assessment tools and collected information were highly heterogeneous across studies. Two studies reported pain treatment strategies and found that only every second person with MSA complaining about pain had received targeted treatment. Conclusions: We found that pain is a frequent, but still under-recognized and undertreated feature of MSA. Further research is needed to improve pain detection and treatment in MSA

po 393 notch3 and cxcr4 cross signalling sustains acute t cell leukaemia progression

Introduction Acute T-cell lymphoblastic leukaemia (T-ALL) is a childhood cancer, characterised by infiltration of immature T-cells in bone marrow. Notch hyperactivation is a major driver of T-ALL development where CXCL12/CXCR4 axis plays an important role in T-ALL maintenance. In thymus the lympho-stromal communication drives progressive maturation of T-cells. Notch receptors regulate T-cell fate choices, dominating early steps of thymocyte maturation. In T-cell differentiation, Notch3, in association with pre-TCR and chemochine receptor CXCR4, govern the transition from double negative (DN) to double positive (DP) thymocytes. Previously, our laboratory demonstrated the lymphomagenic potential of Notch3 by creating a transgenic mouse model (N3-ICtg), characterised by the constitutive activation of the intracellular domain (IC) of Notch3 receptor (N3-IC) in immature thymocytes. In order to investigate the oncogenic cross-talk between Notch3 and CXCR4 in T-ALL progression, we analysed DP T-cells in different lymphoid compartments of N3-ICtg mice. Material and methods Freshly isolated cells from thymus, blood and bone marrow of N3-ICtg and WT mice were analysed by flow cytometry in order to verify the presence of DP T-cells and their cell-surface expression of CXCR4 and Notch3 receptors. Experiments in TALL1, a human T-ALL leukemic CD3 + /CD4 + /CD8 + cell line characterised by the activation of Notch3 and high expression of CXCR4, were also performed. TALL1 cells were treated with γ-secretase inhibitor (GSI) or their gene expression of Notch3 was silenced and then analysed by flow cytometry, RT-PCR and western blot. Statistical interpretation of the results was performed. Results and discussions DP-gated thymocytes obtained by N3-ICtg mice have shown a high co-expression of Notch3 and CXCR4 and a high migratory ability induced by SDF-1. An anomalous percentage representation of these DP T-cells at different ages in circulating blood, spleen and bone morrow may suggest an interaction between CXCR4 and Notch3 in T-ALL cell propagation. Experiments in human TALL1 cell line with Notch3 targeted inhibition suggest a modulated expression of CXCR4 through a β-arrestin1-mediated mechanism. CXCR4-antagonists treatment will further elucidate the molecular crosstalk between the two receptors. Conclusion Notch3 abnormal pathway, through boosting the expression of CXCR4 on cell-surface, may play a role in DP T-cells egress from thymus, and define a possible mechanism of 'pre-leukemic-cells' dissemination

Evaluation of iron overload in nigrosome 1 via quantitative susceptibility mapping as a progression biomarker in prodromal stages of synucleinopathies

Idiopathic rapid eye movement (REM) sleep behavior disorder (iRBD) is a prodromal stage of α-synucleinopathies, such as Parkinson's disease (PD), which are characterized by the loss of dopaminergic neurons in substantia nigra, associated with abnormal iron load. The assessment of presymptomatic biomarkers predicting the onset of neurodegenerative disorders is critical for monitoring early signs, screening patients for neuroprotective clinical trials and understanding the causal relationship between iron accumulation processes and disease development. Here, we used Quantitative Susceptibility Mapping (QSM) and 7T MRI to quantify iron deposition in Nigrosome 1 (N1) in early PD (ePD) patients, iRBD patients and healthy controls and investigated group differences and correlation with disease progression. We evaluated the radiological appearance of N1 and analyzed its iron content in 35 ePD, 30 iRBD patients and 14 healthy controls via T2*-weighted sequences and susceptibility (χ) maps. N1 regions of interest (ROIs) were manually drawn on control subjects and warped onto a study-specific template to obtain probabilistic N1 ROIs. For each subject the N1 with the highest mean χ was considered for statistical analysis. The appearance of N1 was rated pathological in 45% of iRBD patients. ePD patients showed increased N1 χ compared to iRBD patients and HC but no correlation with disease duration, indicating that iron load remains stable during the early stages of disease progression. Although no difference was reported in iron content between iRBD and HC, N1 χ in the iRBD group increases as the disease evolves. QSM can reveal temporal changes in N1 iron content and its quantification may represent a valuable presymptomatic biomarker to assess neurodegeneration in the prodromal stages of PD

O FARMACEUTICO NO CONTEXTO DA ESTRATÉGIA EM SAÚDE DA FAMÍLIA, QUE REALIDADE É ESTA?

The present article reports the pharmacist’s experience on primary health attention, in the context of Brazilian family health strategy. It shows the path to his insertion in the health units, discusses where and how this pharmacist can act in this new proposal and the possible approaches on collective health field. It also brings a reflection about the difficulties during the period of Residence on health family units, the needs of professional education and the different meanings of the medicine to the patient, as well as the integrated work with the multiprofessional team, specially medicine and nutrition.Keywords: pharmacist; family health strategy; health unit.O presente artigo relata a experiência do profissional farmacêutico no âmbito da Atenção Primária, no contexto da Estratégia de Saúde da Família (ESF). Resgata seu histórico até a sua inserção em uma unidade de saúde, discute onde se insere o farmacêutico na proposta de transitoriedade do perfil da assistência e as possíveis abordagens dentro do campo da saúde coletiva. Aponta as dificuldades encontradas durante o período de residência em Unidades de Saúde da Família (USF), as necessidades na formação desse profissional para atuar na Atenção Primária à Saúde e, as diferentes configurações que o medicamento assume perante o indivíduo, bem como o trabalho integrado à equipe multiprofissional, em especial ao profissional médico e ao nutricionista

Ventricular septal defect in a child with Alport syndrome: a case report

<p>Abstract</p> <p>Background</p> <p>Alport syndrome (AS) is a rare inherited disorder characterized by an inflammation of the kidneys and damage to the glomerular capillaries, ultimately leading to renal failure at an early age. To date, rare reports of cardiac involvement in AS have been described, due in the majority of cases to the higher risk of heart conduction abnormalities in these patients, at times requiring implantation of a transcutaneous pacemaker. An increased risk of hypertension is likewise commonly featured.</p> <p>Case presentation</p> <p>We report the case of a 17-year-old female affected by a very severe early form of AS. A previously unreported association of the syndrome with congenital heart disease (CHD), (in this case membranous ventricular septal defect), is also reported. A possible pathophysiological mechanism underlying the concomitant manifestation of these two disorders is suggested. Complications implicated in surgical treatment of CHD are described. Clinical and therapeutic management of AS with cardiovascular involvement are discussed, and a short literature review performed.</p> <p>Conclusions</p> <p>This first report of a cardiovascular association highlights the possible involvement of collagen mutations in the two pathologies. Even when drug-resistance appears to be responsible for the failure to control secondary hypertension in AS, clonidine may represent a safe, effective option in the normalization of high blood pressure.</p