Serum from patients with asthma potentiates macrophage phagocytosis and human mesenchymal stromal cell therapy in experimental allergic asthma.

BACKGROUND/AIMS: Although several studies have demonstrated that mesenchymal stromal cells (MSCs) exhibit beneficial immunomodulatory properties in preclinical models of allergic asthma, effects on airway remodeling have been controversial. Recent evidence has shown that MSCs modify their in vivo immunomodulatory actions depending on the specific inflammatory environment encountered. Accordingly, we assessed whether the therapeutic properties of human mesenchymal stromal cells (hMSCs) could be potentiated by conditioning these cells with serum (hMSC-serum) obtained from patients with asthma and then transplanted in an experimental model of house dust mite (HDM)-induced allergic asthma. METHODS: hMSC and hMSC-serum were administered intratracheally 24 h after the final HDM challenge. hMSC viability and inflammatory mediator production, lung mechanics and histology, bronchoalveolar lavage fluid (BALF) cellularity and biomarker levels, mitochondrial structure and function as well as macrophage polarization and phagocytic capacity were assessed. RESULTS: Serum preconditioning led to: (i) increased hMSC apoptosis and expression of transforming growth factor-ß, interleukin (IL)-10, tumor necrosis factor-α-stimulated gene 6 protein and indoleamine 2,3-dioxygenase-1; (ii) fission and reduction of the intrinsic respiratory capacity of mitochondria; and (iii) polarization of macrophages to M2 phenotype, which may be associated with a greater percentage of hMSCs phagocytosed by macrophages. Compared with mice receiving hMSCs, administration of hMSC-serum led to further reduction of collagen fiber content, eotaxin levels, total and differential cellularity and increased IL-10 levels in BALF, improving lung mechanics. hMSC-serum promoted greater M2 macrophage polarization as well as macrophage phagocytosis, mainly of apoptotic hMSCs. CONCLUSIONS: Serum from patients with asthma led to a greater percentage of hMSCs phagocytosed by macrophages and triggered immunomodulatory responses, resulting in further reductions in both inflammation and remodeling compared with non-preconditioned hMSCs.

Proteomics profile of mesenchymal stromal cells and extracellular vesicles in normoxic and hypoxic conditions.

BACKGROUND AIMS: Although bone marrow-derived mesenchymal stromal cells (MSCs) have demonstrated success in pre-clinical studies, they have shown only mild therapeutic effects in clinical trials. Hypoxia pre-conditioning may optimize the performance of bone marrow-derived MSCs because it better reflects the physiological conditions of their origin. It is not known whether changes in the protein profile caused by hypoxia in MSCs can be extended to the extracellular vesicles (EVs) released from them. The aim of this study was to evaluate the proteomics profile of MSCs and their EVs under normoxic and hypoxic conditions. METHODS: Bone marrow-derived MSCs were isolated from six healthy male Wistar rats. After achieving 80% confluence, MSCs were subjected to normoxia (MSC-Norm) (21% oxygen, 5% carbon dioxide, 74% nitrogen) or hypoxia (MSC-Hyp) (1% oxygen, 5% carbon dioxide, 94% nitrogen) for 48 h. Cell viability and oxygen consumption rate were assessed. EVs were extracted from MSCs for each condition (EV-Norm and EV-Hyp) by ultracentrifugation. Total proteins were isolated from MSCs and EVs and prepared for mass spectrometry. EVs were characterized by nanoparticle tracking analysis. Proteomics data were analyzed by PatternLab 4.0, Search Tool for the Retrieval of Interacting Genes/Proteins, Gene Ontology, MetaboAnalyst and Reactome software. RESULTS: Cell viability was higher in MSC-Hyp than MSC-Norm (P = 0.007). Basal respiration (P = 0.001), proton leak (P = 0.004) and maximal respiration (P = 0.014) were lower in MSC-Hyp than MSC-Norm, and no changes in adenosine triphosphate-linked and residual respiration were observed. The authors detected 2177 proteins in MSC-Hyp and MSC-Norm, of which 147 were identified in only MSC-Hyp and 512 were identified in only MSC-Norm. Furthermore, 718 proteins were identified in EV-Hyp and EV-Norm, of which 293 were detected in only EV-Hyp and 30 were detected in only EV-Norm. Both MSC-Hyp and EV-Hyp showed enrichment of pathways and biological processes related to glycolysis, the immune system and extracellular matrix organization. CONCLUSIONS: MSCs subjected to hypoxia showed changes in their survival and metabolic activity. In addition, MSCs under hypoxia released more EVs, and their content was related to expression of regulatory proteins of the immune system and extracellular matrix organization. Because of the upregulation of proteins involved in glycolysis, gluconeogenesis and glucose uptake during hypoxia, production of reactive oxygen species and expression of immunosuppressive properties may be affected.

Infection of Endothelial Cells by Dengue Virus Induces ROS Production by Different Sources Affecting Virus Replication, Cellular Activation, Death and Vascular Permeability.

Exacerbated inflammatory response and altered vascular function are hallmarks of dengue disease. Reactive oxygen species (ROS) production has been associated to endothelial barrier disturbance and microvascular alteration in distinct pathological conditions. Increased ROS has been reported in in vitro models of dengue virus (DENV) infection, but its impact for endothelial cell physiology had not been fully investigated. Our group had previously demonstrated that infection of human brain microvascular endothelial cells (HBMEC) with DENV results in the activation of RNA sensors and production of proinflammatory cytokines, which culminate in cell death and endothelial permeability. Here, we evaluated the role of mitochondrial function and NADPH oxidase (NOX) activation for ROS generation in HBMEC infected by DENV and investigated whether altered cellular physiology could be a consequence of virus-induced oxidative stress. DENV-infected HBMECs showed a decrease in the maximal respiratory capacity and altered membrane potential, indicating functional mitochondrial alteration, what might be related to mtROS production. Indeed, mtROS was detected at later time points after infection. Specific inhibition of mtROS diminished virus replication, cell death, and endothelial permeability, but did not affect cytokine production. On the other hand, inhibition of NOX-associated ROS production decreased virus replication and cell death, as well as the secretion of inflammatory cytokines, including IL-6, IL-8, and CCL5. These results demonstrated that DENV replication in endothelial cells induces ROS production by different pathways, which impacts biological functions that might be relevant for dengue pathogenesis. Those data also indicate oxidative stress events as relevant therapeutical targets to avoid vascular permeability, inflammation, and neuroinvasion during DENV infection.

Mitochondria-Rich Fraction Isolated From Mesenchymal Stromal Cells Reduces Lung and Distal Organ Injury in Experimental Sepsis.

OBJECTIVES: To ascertain whether systemic administration of mitochondria-rich fraction isolated from mesenchymal stromal cells would reduce lung, kidney, and liver injury in experimental sepsis. DESIGN: Animal study. SETTING: Laboratory investigation. SUBJECTS: Sixty C57BL/6 male mice. INTERVENTIONS: Sepsis was induced by cecal ligation and puncture; sham-operated animals were used as control. At 24 hours after surgery, cecal ligation and puncture and Sham animals were further randomized to receive saline or mitochondria-rich fraction isolated from mesenchymal stromal cells (3 × 106) IV. At 48 hours, survival, peritoneal bacterial load, lung, kidney, and liver injury were analyzed. Furthermore, the effects of mitochondria on oxygen consumption rate and reactive oxygen species production of lung epithelial and endothelial cells were evaluated in vitro. MEASUREMENTS AND MAIN RESULTS: In vitro exposure of lung epithelial and endothelial cells from cecal ligation and puncture animals to mitochondria-rich fraction isolated from mesenchymal stromal cells restored oxygen consumption rate and reduced total reactive oxygen species production. Infusion of exogenous mitochondria-rich fraction from mesenchymal stromal cells (mitotherapy) reduced peritoneal bacterial load, improved lung mechanics and histology, and decreased the expression of interleukin-1ß, keratinocyte chemoattractant, indoleamine 2,3-dioxygenase-2, and programmed cell death protein 1 in lung tissue, while increasing keratinocyte growth factor expression and survival rate in cecal ligation and puncture-induced sepsis. Mitotherapy also reduced kidney and liver injury, plasma creatinine levels, and messenger RNA expressions of interleukin-18 in kidney, interleukin-6, indoleamine 2,3-dioxygenase-2, and programmed cell death protein 1 in liver, while increasing nuclear factor erythroid 2-related factor-2 and superoxide dismutase-2 in kidney and interleukin-10 in liver. CONCLUSIONS: Mitotherapy decreased lung, liver, and kidney injury and increased survival rate in cecal ligation and puncture-induced sepsis.

Human mesenchymal stem cell therapy promotes retinal ganglion cell survival and target reconnection after optic nerve crush in adult rats.

BACKGROUND: Optic-nerve injury results in impaired transmission of visual signals to central targets and leads to the death of retinal ganglion cells (RGCs) and irreversible vision loss. Therapies with mesenchymal stem cells (MSCs) from different sources have been used experimentally to increase survival and regeneration of RGCs. METHODS: We investigated the efficacy of human umbilical Wharton's jelly-derived MSCs (hWJ-MSCs) and their extracellular vesicles (EVs) in a rat model of optic nerve crush. RESULTS: hWJ-MSCs had a sustained neuroprotective effect on RGCs for 14, 60, and 120 days after optic nerve crush. The same effect was obtained using serum-deprived hWJ-MSCs, whereas transplantation of EVs obtained from those cells was ineffective. Treatment with hWJ-MSCs also promoted axonal regeneration along the optic nerve and reinnervation of visual targets 120 days after crush. CONCLUSIONS: The observations showed that this treatment with human-derived MSCs promoted sustained neuroprotection and regeneration of RGCs after optic nerve injury. These findings highlight the possibility to use cell therapy to preserve neurons and to promote axon regeneration, using a reliable source of human MSCs.

Imunoterapia com células CAR-T como nova perspectiva de tratamento da leucemia linfoblástica aguda recidivada/refratária / Immunotherapy with CAR-T cells as a new treatment perspective for relapsed/refractory acute lymphoblastic leukemia

Introdução: O tratamento da leucemia linfoblástica aguda (LLA) atualmente baseia-se em quimioterapia e/ou transplante de células tronco hematopoiéticas; entretanto, uma nova terapia vem se tornando promissora: a imunoterapia com células T modificadas geneticamente que expressam um receptor de antígeno quimérico (CAR-T) visando antígenos específicos presente em blastos de LLA, gerando resultados promissores em crianças e adultos com doença recidivada e refratária (r/r). Objetivo: Discorrer sobre a LLA e descrever a imunoterapia com CAR-T, como inovação terapêutica no tratamento da LLA de linhagem B. Método: Foi realizada uma revisão bibliográfica por meio de publicações indexadas nas bases de dados Scielo e Pubmed, utilizando os descritores: leucemia linfoblástica aguda de células B; células CAR-T; receptores de antígeno quimérico, recidivados/refratários; imunoterapia. Resultados: As altas taxas de remissão completa (42% até 100%) e parcial (28,5%) da LLA (r/r) tratadas com CAR-T, possibilitam um aumento considerável da sobrevida geral comparado a outros tratamentos convencionais. Efeitos desfavoráveis, tais como síndrome da liberação de citocinas (CRS) (0 até 90%) e neurotoxicidade (NT) (0 até 29%) podem ser vistos, sendo manejáveis, não prejudicando o desfecho do tratamento. Conclusão: A LLA é uma doença grave, de difícil tratamento e prognóstico reservado. A imunoterapia vêm se mostrando promissora à essa enfermidade, principalmente em casos de doença r/r se mostrado uma ferramenta poderosa que permite o foco específico de células malignas por meio de engenharia de células T

Introduction: The treatment of acute lymphoblastic leukemia (ALL) is currently based on chemotherapy and/or hematopoietic stem cell transplantation; however, a new therapy is becoming promising: immunotherapy with genetically modified T cells that express a chimeric antigen receptor (CAR-T) targeting specific antigens present on ALL blasts, reaching promising results in children and adults with relapsed and refractory disease (r/r). Objective: To discuss ALL and describe immunotherapy with CAR-T as a therapeutic innovation in the treatment of B-lineage ALL. Method: A literature review was carried out through publications indexed in the Scielo and Pubmed databases, using the following descriptors: B-cell acute lymphoblastic leukemia; CAR-T cells; chimeric antigen receptors, relapsed/refractory; immunotherapy. Results: The high rates of complete (42% to 100%) and partial remission (28.5%) of ALL (r/r) treated with CAR-T allows a considerable increase in overall survival compared to other conventional treatments. Unfavorable effects such as cytokine release syndrome (CRS) (0 to 90%) and neurotoxicity (NT) (0 to 29%) can be seen, being manageable, not impairing the treatment outcome. Conclusion: ALL is a serious disease, with a difficult treatment and poor prognosis. Immunotherapy has shown benefits for this disease, especially in cases of r/r ALL, showing itself to be a powerful tool that allows the specific focus of malignant cells through T cell engineering.

Human Wharton's jelly mesenchymal stem cells protect neural cells from oxidative stress through paracrine mechanisms.

AIM: Mesenchymal stem cells (MSCs) have neuroprotective and immunomodulatory properties, which are partly mediated by extracellular vesicles (EVs) secretion. We aimed to evaluate the effects of human Wharton's jelly-derived MSCs (WJ-MSCs) and their EVs on rat hippocampal cultures subjected to hydrogen peroxide (H2O2). MATERIALS & METHODS: Hippocampal dissociated cultures were either co-cultured with WJ-MSCs or treated with their EVs prior to H2O2 exposure and reactive oxygen species levels and cell viability were evaluated. RESULTS: Coculture with WJ-MSCs or pre-incubation with EVs prior to the insult reduced reactive oxygen species after H2O2 exposure. Cell viability was improved only when coculture was maintained following the insult, while EVs did not significantly improve cell viability. CONCLUSION: WJ-MSCs have potential antioxidant and neuroprotective effects on hippocampal cultures which might be partially mediated by EVs.

Association between magnesium, selenium and zinc consumption and lipid profile of brazilian adolescents / Asociación entre el consumo de magnesio, selenio y zinc y el perfil lipídico de los adolescentes brasileños

ABSTRACT The study aims to evaluate the association between inadequate consumption of antioxidant minerals and plasma lipoprotein concentrations in adolescents. We conducted a cross-sectional study that evaluated sociodemographic and anthropometric data, information on intake of magnesium, selenium and zinc and lipid profile. Student's t-test was used to compare means between the groups and logistic regression to verify the strength of the association between the independent variables and lipid profile. Inadequate zinc consumption was associated with a higher chance of low HDL-c levels and lower chance of hypertriglyceridemia and high LDL-c levels. Inadequate selenium intake was associated with a lower chance of high total cholesterol and of high triglyceride concentrations and a higher chance of low HDL-c levels. Inadequate magnesium consumption was associated with a higher chance of high cholesterol and triglyceride levels, a lower chance of high LDL-c levels and with a higher chance of low HDL-c. We observed an association between inadequate consumption of magnesium, zinc and selenium and changes in the lipid profile of adolescents.

RESUMEN El estudio tiene como objetivo evaluar la asociación entre el consumo inadecuado de minerales antioxidantes y las concentraciones plasmáticas de lipoproteínas en adolescentes. Estudio transversal que evaluó datos sociodemográficos y antropométricos, información sobre ingesta de magnesiom selenio y zinc y perfil lipídico. Se utilizó la prueba t de Student para comparar medias entre los grupos y regresión logística para verificar la fuerza de la asociación entre las variables independientes y el perfil lipídico. El consumo inadecuado de zinc se asoció con una mayor probabilidad de niveles bajos de HDL-c y una menor probabilidad de hipertrigliceridemia y niveles altos de LDL-c. La ingesta inadecuada de selenio se asoció con una menor probabilidad de colesterol total alto y de altas concentraciones de triglicéridos y una mayor probabilidad de niveles bajos de HDL-c. El consumo inadecuado de magnesio se asoció con una mayor probabilidad de niveles altos de colesterol y triglicéridos, una menor probabilidad de niveles altos de LDL-c y una mayor probabilidad de niveles bajos de HDL-c. El estudio muestra una asociación entre el consumo inadecuado de magnesio, zinc y selenio y los cambios en el perfil lipídico de los adolescentes.

Effects of a combinatorial treatment with gene and cell therapy on retinal ganglion cell survival and axonal outgrowth after optic nerve injury.

After an injury, axons in the central nervous system do not regenerate over large distances and permanently lose their connections to the brain. Two promising approaches to correct this condition are cell and gene therapies. In the present work, we evaluated the neuroprotective and neuroregenerative potential of pigment epithelium-derived factor (PEDF) gene therapy alone and combined with human mesenchymal stem cell (hMSC) therapy after optic nerve injury by analysis of retinal ganglion cell survival and axonal outgrowth. Overexpression of PEDF by intravitreal delivery of AAV2 vector significantly increased Tuj1-positive cells survival and modulated FGF-2, IL-1ß, Iba-1, and GFAP immunostaining in the ganglion cell layer (GCL) at 4 weeks after optic nerve crush, although it could not promote axonal outgrowth. The combination of AAV2.PEDF and hMSC therapy showed a higher number of Tuj1-positive cells and a pronounced axonal outgrowth than unimodal therapy after optic nerve crush. In summary, our results highlight a synergistic effect of combined gene and cell therapy relevant for future therapeutic interventions regarding optic nerve injury.

Consulta de enfermagem em puericultura à criança haitiana: dificuldades e possibilidades / Consulta de enfermería en puericultura al niño haitiano: dificultades y posibilidades / Nursing consultation in childcare to haitian children: difficulties and possibilities

Objetivo: identificar as dificuldades vivenciadas pelos enfermeiros na Consulta de Enfermagem em puericultura à criança haitiana na Atenção Primária à Saúde e as possibilidades para superar esses desafios Método: estudo exploratório, descritivo com abordagem qualitativa, realizado com dez enfermeiros por meio de entrevista realizada em julho de 2018. Submeteram-se os depoimentos à Análise de Conteúdo. Resultados: a análise dos dados deu origem a duas categorias: Dificuldades na intercomunicação entre enfermeiros e famílias de crianças haitianas e Dificuldades culturais relacionadas ao cuidado da criança haitiana. Conclusão: os resultados revelam dificuldades na realização da Consulta de Enfermagem às crianças, especialmente relacionadas à intercomunicação, pela língua falada pelos haitianos, e pelas questões culturais que permeiam e embasam o cuidado à criança pelas famílias. Para superar as dificuldades os enfermeiros indicam o uso da comunicação não verbal, visita domiciliar para conhecer o contexto e melhorar a confiança e o vínculo com as famílias.

Objetivo: identificar las dificultades vividas por los enfermeros en la Consulta de Enfermería en puericultura al niño haitiano en la Atención Primaria a la Salud y las posibilidades para superar esos desafíos Método: estudio exploratorio, descriptivo con enfoque cualitativo, realizado con diez enfermeros por medio de una entrevista realizada en julio de 2018. Las declaraciones se sometieron al Análisis de Contenido. Resultados: dos categorias surgieron del análisis de los datos: Dificultades en la intercomunicación entre enfermeros y familias de niños haitianos y Dificultades culturales relacionadas al cuidado del niño haitiano. Conclusión: Los resultados muestran dificultades en la realización de la Consulta de Enfermería a los niños, especialmente relacionadas a la intercomunicación, por edioma hablado por los haitianos, y por las cuestiones culturales permeando y basando el cuidado al niño por las familias. Para superar las dificultades, los enfermeros indican el uso de la comunicación no verbal, visita domiciliaria para conocer el contexto y mejorar la confianza y el vínculo con las familias.

Objective: To identify the difficulties lived by the nurses during the Nursing Consultation in childcare to Haitian children in the Primary Health Care and the possibilities to overcome these challenges. Method: this is an exploratory, descriptive study with a qualitative approach, conducted with ten nurses through an interview held in July 2018. The testimonies were submitted to Content Analysis. Results: From the data analysis, two categories emerged: Difficulties in the intercommunication between nurses and families of Haitian children and Cultural difficulties for the care of Haitian children. Conclusion: the results reveal difficulties in carrying out the Nursing Consultation for children, especially related to intercommunication, due to the language spoken by Haitians, and due to the cultural issues that permeate and support the care of children by families. To overcome the difficulties, nurses indicate the use of non-verbal communication, home visits to get to know the context and improve trust and bond with the families.

Extracellular vesicles derived from human Wharton's jelly mesenchymal stem cells protect hippocampal neurons from oxidative stress and synapse damage induced by amyloid-ß oligomers.

BACKGROUND: Mesenchymal stem cells (MSCs) have been explored as promising tools for treatment of several neurological and neurodegenerative diseases. MSCs release abundant extracellular vesicles (EVs) containing a variety of biomolecules, including mRNAs, miRNAs, and proteins. We hypothesized that EVs derived from human Wharton's jelly would act as mediators of the communication between hMSCs and neurons and could protect hippocampal neurons from damage induced by Alzheimer's disease-linked amyloid beta oligomers (AßOs). METHODS: We isolated and characterized EVs released by human Wharton's jelly mesenchymal stem cells (hMSC-EVs). The neuroprotective action of hMSC-EVs was investigated in primary hippocampal cultures exposed to AßOs. RESULTS: hMSC-EVs were internalized by hippocampal cells in culture, and this was enhanced in the presence of AßOs in the medium. hMSC-EVs protected hippocampal neurons from oxidative stress and synapse damage induced by AßOs. Neuroprotection by hMSC-EVs was mediated by catalase and was abolished in the presence of the catalase inhibitor, aminotriazole. CONCLUSIONS: hMSC-EVs protected hippocampal neurons from damage induced by AßOs, and this was related to the transfer of enzymatically active catalase contained in EVs. Results suggest that hMSC-EVs should be further explored as a cell-free therapeutic approach to prevent neuronal damage in Alzheimer's disease.

Leporacarus gibbus (Acari: listrophoridae) in a naturally-infested domestic rabbit: first report of its occurrence in the state of Espírito Santo, Brazil / Leporacarus gibbus (Acari: listrophoridae) em um coelho doméstico naturalmente infestado: primeiro relato de sua ocorrência no estado do Espírito Santo, Brasil

This study aimed to report the first occurrence of Leporacarus gibbus in a domestic rabbit (Oryctolagus cuniculus) from the state of Espírito Santo, Brazil. The two-year-old male rabbit came from a rural area and was admitted to the Veterinary Hospital Professor Ricardo Alexandre Hippler with intense pruritus throughout the body, severe hair loss, crustal lesions on the outer surface of the ears, and desquamation mainly in the dorsal region. A skin scraping and trichogram were performed. In the microscopic evaluation of the hairs obtained from the dorsal region, mites with morphology compatible with L. gibbus were observed. Cheyletiella parasitovorax and Psoropotes cuniculi were also detected in the same region. Treatment with ivermectin (0.4 mg/kg) was prescribed with three applications every 14 days, over a period of six weeks. This report presents the first description of the occurrence of L. gibbus in the state of Espírito Santo concomitant with poly-infestation of C. parasitovorax and P. cuniculi, and it is hoped that this will provide a research tool for future work in the region.(AU)

Este trabalho teve como objetivo relatar a primeira ocorrência de Leporacarus gibbus em um coelho doméstico (Oryctolagus cuniculus) do estado do Espírito Santo, Brasil. O coelho macho de dois anos de idade veio de uma área rural e foi internado no Hospital Veterinário Professor Ricardo Alexandre Hippler com intenso prurido em todo o corpo, severa queda de cabelo, lesões crustais na superfície externa das orelhas e descamação principalmente na região dorsal. A raspagem da pele e o tricograma foram realizados. Na avaliação microscópica dos pêlos obtidos na região dorsal, observaram-se ácaros com morfologia compatível com L. gibbus. Cheyletiella parasitovorax e Psoropotes cuniculi também foram detectados na mesma região. O tratamento com ivermectina (0,4 mg / kg) foi prescrito com três aplicações a cada 14 dias, durante um período de seis semanas. Este relato apresenta a primeira descrição da ocorrência de L. gibbus no estado do Espírito Santo concomitante à poliinfestação de C. parasitovorax e P. cuniculi, e espera-se que esta seja uma ferramenta de pesquisa para futuros trabalhos na região.(AU)

Mesenchymal stem cells and cell-derived extracellular vesicles protect hippocampal neurons from oxidative stress and synapse damage induced by amyloid-ß oligomers.

Alzheimer's disease (AD) is a disabling and highly prevalent neurodegenerative condition, for which there are no effective therapies. Soluble oligomers of the amyloid-ß peptide (AßOs) are thought to be proximal neurotoxins involved in early neuronal oxidative stress and synapse damage, ultimately leading to neurodegeneration and memory impairment in AD. The aim of the current study was to evaluate the neuroprotective potential of mesenchymal stem cells (MSCs) against the deleterious impact of AßOs on hippocampal neurons. To this end, we established transwell cocultures of rat hippocampal neurons and MSCs. We show that MSCs and MSC-derived extracellular vesicles protect neurons against AßO-induced oxidative stress and synapse damage, revealed by loss of pre- and postsynaptic markers. Protection by MSCs entails three complementary mechanisms: 1) internalization and degradation of AßOs; 2) release of extracellular vesicles containing active catalase; and 3) selective secretion of interleukin-6, interleukin-10, and vascular endothelial growth factor to the medium. Results support the notion that MSCs may represent a promising alternative for cell-based therapies in AD.

A panel of clinical and neuropathological features of cerebrovascular disease through the novel neuroimaging methods / Um painel de características clínicas e neuropatológicas da doença cerebrovascular através dos novos métodos de neuroimagem

ABSTRACT. The last decade has witnessed substantial progress in acquiring diagnostic biomarkers for the diagnostic workup of cerebrovascular disease (CVD). Advanced neuroimaging methods not only provide a strategic contribution for the differential diagnosis of vascular dementia (VaD) and vascular cognitive impairment (VCI), but also help elucidate the pathophysiological mechanisms ultimately leading to small vessel disease (SVD) throughout its course. Objective: In this review, the novel imaging methods, both structural and metabolic, were summarized and their impact on the diagnostic workup of age-related CVD was analysed. Methods: An electronic search between January 2010 and 2017 was carried out on PubMed/MEDLINE, Institute for Scientific Information Web of Knowledge and EMBASE. Results: The use of full functional multimodality in simultaneous Magnetic Resonance (MR)/Positron emission tomography (PET) may potentially improve the clinical characterization of VCI-VaD; for structural imaging, MRI at 3.0 T enables higher-resolution scanning with greater imaging matrices, thinner slices and more detail on the anatomical structure of vascular lesions. Conclusion: Although the importance of most of these techniques in the clinical setting has yet to be recognized, there is great expectancy in achieving earlier and more refined therapeutic interventions for the effective management of VCI-VaD.

RESUMO. A última década vem testemunhando avanços substanciais na aquisição de marcadores biológicos para o diagnóstico da doença cerebrovascular (DCV). Os métodos de neuroimagem avançados não apenas fornecem uma contribuição estratégica para o diagnóstico diferencial do comprometimento cognitivo vascular (VCI) e demência vascular (VaD), mas contribuem substancialmente na elucidação dos mecanismos fisiopatológicos relacionados à doença de vasos pequenos (SVD) e sua progressão clínica. Objetivo: Nesta revisão, métodos de imagem estruturais e metabólicos foram descritos e sua importância diagnóstica analisada, particularmente na investigação da CVD relacionada ao envelhecimento. Métodos: uma pesquisa eletrônica de janeiro de 2010 a 2017 foi realizada através do PubMed/MEDLINE, do Instituto de Informação Científica Web of Knowledge e da EMBASE. Resultados: O emprego de estudos de multimodalidade plenamente funcional com Ressonância Magnética (MR)/ Tomografia por Emissão de Pósitrons (PET) representa uma janela para a caracterização clínica mais detalhada da VCI-VaD; com relação à neuroimagem estrutural, a ressonância magnética em 3,0 T vem permitindo varreduras com maior resolução e matrizes de imagem mais elevadas, cortes mais delgados e maior detalhamento anatômico das lesões vasculares. Conclusão: Embora a importância da maior parte dessas técnicas no cenário clínico aguarde reconhecimento, há uma grande expectativa de que o seu uso favoreça intervenções terapêuticas progressivamente mais precoces e refinadas para o gerenciamento efetivo do VCI-VaD.

Neuroimaging Findings Related to Behavioral Disturbances in Alzheimer's Disease: A Systematic Review.

BACKGROUND: Behavioral and psychological symptoms of dementia (BPSD) associated with Alzheimer's Disease (AD) have been linked to structural and functional alterations in fronto-temporal circuits and cortical abnormalities. However, little is known on how specific volumetric and functional brain changes may be associated with the frequency, severity and pattern of BPSD. METHODS: A systematic review of the literature regarding neuroimaging and BPSD changes in AD was performed through Pubmed/Medline, ISI, and EMBASE electronic databases from January 2000 to May 2015. Eligible references (n=40) included clinical studies in which structural or functional neuroimaging assessment was performed in AD subjects presenting BPSD features. RESULTS: BPSD symptoms, particularly apathy and psychosis have been associated in most of studies with either volume reductions or decreased metabolism in the prefrontal cortex (orbital and dorsolateral portions), anterior cingulate, insula and temporal lobes (middle portion). WM lacunes associated with AD progression have been associated with depressive symptoms. CONCLUSION: The sum of evidence highlights the importance of BPSD-related imaging findings for the understanding of the non-cognitive symptom spectrum in AD. Results suggest that structural and functional changes in fronto-limbic areas may lead to emotional deregulation and symptom unawareness. As these findings may be present early on the AD clinical course, they may have a relevance for the development of imaging markers that could be used in diagnosis, disease monitoring and prediction of therapeutic response.

A panel of clinical and neuropathological features of cerebrovascular disease through the novel neuroimaging methods.

The last decade has witnessed substantial progress in acquiring diagnostic biomarkers for the diagnostic workup of cerebrovascular disease (CVD). Advanced neuroimaging methods not only provide a strategic contribution for the differential diagnosis of vascular dementia (VaD) and vascular cognitive impairment (VCI), but also help elucidate the pathophysiological mechanisms ultimately leading to small vessel disease (SVD) throughout its course. OBJECTIVE: In this review, the novel imaging methods, both structural and metabolic, were summarized and their impact on the diagnostic workup of age-related CVD was analysed. Methods: An electronic search between January 2010 and 2017 was carried out on PubMed/MEDLINE, Institute for Scientific Information Web of Knowledge and EMBASE. RESULTS: The use of full functional multimodality in simultaneous Magnetic Resonance (MR)/Positron emission tomography (PET) may potentially improve the clinical characterization of VCI-VaD; for structural imaging, MRI at 3.0 T enables higher-resolution scanning with greater imaging matrices, thinner slices and more detail on the anatomical structure of vascular lesions. CONCLUSION: Although the importance of most of these techniques in the clinical setting has yet to be recognized, there is great expectancy in achieving earlier and more refined therapeutic interventions for the effective management of VCI-VaD.

A última década vem testemunhando avanços substanciais na aquisição de marcadores biológicos para o diagnóstico da doença cerebrovascular (DCV). Os métodos de neuroimagem avançados não apenas fornecem uma contribuição estratégica para o diagnóstico diferencial do comprometimento cognitivo vascular (VCI) e demência vascular (VaD), mas contribuem substancialmente na elucidação dos mecanismos fisiopatológicos relacionados à doença de vasos pequenos (SVD) e sua progressão clínica. OBJETIVO: Nesta revisão, métodos de imagem estruturais e metabólicos foram descritos e sua importância diagnóstica analisada, particularmente na investigação da CVD relacionada ao envelhecimento. Métodos: uma pesquisa eletrônica de janeiro de 2010 a 2017 foi realizada através do PubMed/MEDLINE, do Instituto de Informação Científica Web of Knowledge e da EMBASE. RESULTADOS: O emprego de estudos de multimodalidade plenamente funcional com Ressonância Magnética (MR)/ Tomografia por Emissão de Pósitrons (PET) representa uma janela para a caracterização clínica mais detalhada da VCI-VaD; com relação à neuroimagem estrutural, a ressonância magnética em 3,0 T vem permitindo varreduras com maior resolução e matrizes de imagem mais elevadas, cortes mais delgados e maior detalhamento anatômico das lesões vasculares. CONCLUSÃO: Embora a importância da maior parte dessas técnicas no cenário clínico aguarde reconhecimento, há uma grande expectativa de que o seu uso favoreça intervenções terapêuticas progressivamente mais precoces e refinadas para o gerenciamento efetivo do VCI-VaD.

White matter abnormalities in the fornix are linked to cognitive performance in SZ but not in BD disorder: An exploratory analysis with DTI deterministic tractography.

BACKGROUND: In psychosis, white matter (WM) microstructural changes have been detected previously; however, direct comparisons of findings between bipolar (BD) and schizophrenia (SZ) patients are scarce. In this study, we employed deterministic tractography to reconstruct WM tracts in BD and SZ patients. METHODS: Diffusion tensor imaging (DTI) data was carried out with n=32 euthymic BD type I patients, n=26 SZ patients and 30 matched healthy controls. Deterministic tractography using multiple indices of diffusion (fractional anisotropy (FA), tract volume (Vol), tract length (Le) and number of tracts (NofT)) were obtained from the fornix, the cingulum, the anterior thalamic radiation, and the corpus callosum bilaterally. RESULTS: We showed widespread WM microstructural changes in SZ, and changes in the corpus callosum, the left cingulum and the fornix in BD. Fornix fiber tracking scores were associated with cognitive performance in SZ, and with age and age at disease onset in the BD patient group. LIMITATIONS: Although the influence of psychopharmacological drugs as biasing variables on morphological alterations has been discussed for SZ and BD, we did not observe a clear influence of drug exposure on our findings. CONCLUSIONS: These results confirm the assumption that SZ patients have more severe WM changes than BD patients. The findings also suggest a major role of WM changes in the fornix as important fronto-limbic connections in the etiology of cognitive symptoms in SZ, but not in BD.

Structural neuroimaging findings in major depressive disorder throughout aging: a critical systematic review of prospective studies.

Clinical manifestations of major depressive disorder (MDD) have been linked to structural and functional alterations in fronto-limbic circuits and white matter microstructural abnormalities. However, little is known about how brain pathological changes in volume and microstructure are related to illness progression throughout aging, including course deterioration and treatment response. A comprehensive review of the literature regarding midlife- and late-onset MDD was performed through PubMed/Medline, ISI, and EMBASE electronic databases from January 2000 to May 2014. Eligible references included prospective studies in which structural neuroimaging assessments were performed in MDD samples. The course of MDD may be associated with brain aging modifications, including hippocampal, amigdalar and frontal volume reductions. White matter changes associated with MDD progression have been reported in the corpus callosum, frontal and temporal regions and may be associated with poorer response to treatment. The data suggest that both cortical and subcortical alterations may interact along the progression of MDD. Further knowledge brought by neuroimaging studies, through the integration of multimodal techniques, may help to improve the accuracy of diagnosis, disease monitoring and treatment response in MDD.

Genotype and expression of the enhanced gree fluorescent protein in LEW-Tg (EGFP) F455.5/Rrrc rats / Genótipo e expressão da proteína fluorescente verde melhorada em ratos da linhagem LEW-Tg (EGFP) F455.5/Rrrc

The Green fluorescent protein (GFP) was first described after being extracted from Aequorea victoria in 1987; Since then, GFP and its derivatives have been widely used in several experiments as cell and protein marker. In the present study it was verified the genotype of the offspring from crosses between heterozygote Lewis LEW-Tg (EGFP) F455.5/Rrrc rats and analyzed the expression of the enhanced green fluorescent protein (EGFP) in different cell types and genotypes. The genotype of the offspring was assessed by PCR and analysis of EGFP expression in different cells and genotypes, including mesenchymal stem cells (MSC) derived from adipose tissue and calvarial osteoblast cells. Expression of EGFP was verified by flow cytometry, fluorescence microscopy, and immunostaining. Through these methods, it was identified the genotypes of the offspring and determined the levels of expression of EGFP in two cell types. A difference in expression between the (EGFP +/+) and (EGFP +/-) genotypes was also observed in addition to the presence of autofluorescence. Further studies on the natural fluorescence of cells with the (EGFP +/-) genotype and that induced by presence of the EGFP are necessary.

A proteína fluorescente verde (GFP) foi descrita pela primeira vez após ter sido extraída de Aequorea victoria em 1987. Desde então, a GFP e seus derivados têm sido amplamente utilizados em várias experiências como marcador celular e de proteínas. O objetivo do presente estudo foi o de verificar o genótipo dos descendentes de cruzamentos entre ratos Lewis LEW-Tg (EGFP) F455.5/Rrrc heterozigotos e de analisar a expressão da proteína fluorescente verde melhorada (EGFP) em diferentes tipos celulares e genótipos. O genótipo da descendência foi avaliado por PCR e pela análise da expressão da EGFP em diferentes células e genótipos, incluindo-se as células-tronco mesenquimais (MSC) derivadas de tecido adiposo e de osteoblastos de calvária. A expressão da EGFP foi verificada por citometria de fluxo, microscopia de fluorescência e imunocoloração. Foram, identificados os genótipos da descendência e determinados os níveis de expressão de EGFP em dois tipos de células. Foi também constatada uma diferença de expressão entre os genótipos (EGFP +/+) e (EGFP +/-) além da presença de autofluorescência. Mais estudos são necessários para esclarecer a fluorescência natural de células com o genótipo (EGFP +/-) e aquela induzida pela presença da EGFP.

Effect of a three-dimensional chitosan porous scaffold on the differentiation of mesenchymal stem cells into chondrocytes.

Cartilage tissue has a poor capacity for self-repair, especially in the case of severe cartilage damage due to trauma or age-related degeneration. Cell-based tissue engineering using scaffolds has provided an option for the repair of cartilage tissue. The present work demonstrates that a three-dimensional (3D) chitosan scaffold increases the efficiency of the adhesion and differentiation of mesenchymal stem cells (MSCs) after the addition of a chondrogenic medium. These culture conditions promoted MSC differentiation into chondrocytes during the first 9 weeks of monolayer or 3D culture in a scaffold composed of chitosan or chitosan/gelatin. The results demonstrated that a chitosan scaffold caused a reduction in alkaline phosphatase production and an increase in the collagen concentration indicating phenotypic changes in the cells. In support of these results, the production of collagen type II by the MSCs cultured in the chitosan scaffold increased after 3 weeks of culture, indicating the beginning of differentiation. However, the addition of gelatin to the chitosan scaffold did not improve on the results obtained with chitosan alone. These results suggest that this 3D chitosan scaffold is a promising candidate for biomaterial implants designed to promote MSC colonization and has applications in regenerative medicine.