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.

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.

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.

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.

Prótese protocolo com carga imediata em sítio infectado / Implant-supported fixed prosthesis with immediate loading technique in infected site: case report

Os implantes osseointegráveis possuem grande previsibilidade, porém, para se obter os índices de sucesso descritos na literatura para a técnica de carga imediata, vários fatores devem ser observados, como por exemplo, a qualidade óssea da área receptora, a estabilidade primária dos implantes e a presença de infecção no sítio cirúrgico. Atualmente, os pacientes têm buscado cada vez mais a recuperação da função mastigatória e estética, da forma mais eficaz, cômoda e em um menor tempo de tratamento. Isso pode se concretizar graças aos procedimentos de carga imediata. O objetivo desse trabalho foi abordar a técnica de carga imediata em sítio infectado, mediante relato de um caso clínico. Concluiu-se que esta técnica apresenta como principal vantagem a redução do tempo de tratamento. Todo planejamento deve ser previamente aprovado pelo paciente, apresentando-lhe os prós e contras da técnica, diminuindo assim a possibilidade de frustrações no caso de insucesso.

Osseointegrated implants have a large predictability; although, to get the rate of success described in the literature regarding immediate loading, many factors must be observed such as bone quality of the receptor area, implant primary stability, and the presence of infection in the surgical site. Currently, patients seek rehabilitation of chewing and esthetic function in a more effective way along with minimum treatment time. This can be achieved thanks to immediate loading technique. The aim of this work is to describe immediate loading technique in an infected site, using a clinical case. We conclude that this technique has the minor treatment time as its main advantage. However, treatment planning should be previously approved by the patient, presenting advantages and drawbacks, decreasing the rate of frustration in case of failure.

Doença pneumocócica invasiva em crianças e adolescentes soropositivos para HIV / Invasive pneumococcal disease in HIV seropositive children and adolescents

OBJETIVO: Doença Invasiva Pneumocócica (DPI) afeta crianças principalmente menores de 5 anos, idosos e grupos de risco, especialmente pessoas infectadas pelo vírus da Imunodeficiência Humana (HIV). O objetivo deste trabalho foi analisar as doenças pneumocócicas invasivas (DPI) em crianças e adolescentes infectados pelo vírus da imunodeficiência humana (HIV), de acordo com morbiletalidade, sorotipos, sensibilidade à penicilina e ceftriaxona e distribuição de Streptococcus pneumoniae (Sp) sensíveis e resistentes presentes na vacina antipneumocócica conjugada 7-valente, já licenciada. MÉTODOS: Foram identificados 19 casos de DPI entre pacientes HIV soropositivos com idade entre 1 mês e 20 anos hospitalizados de 1993 a 2000. Os dados foram registrados em fichas padronizadas, contendo informações sobre idade, diagnóstico clínico e evolução, sorotipos e perfil de sensibilidade para penicilina e ceftriaxona das cepas de Sp isoladas em cultura. Sp com concentração inibitória mínima < 0,1 mcg/mL foi considerado sensível à penicilina (SpSPn), e as demais cepas como não sensíveis (SpNSPn). RESULTADOS: Dos 19 casos de DPI em HIV soropositivos, 16 (84 por cento) tinham pneumonia e três (16 por cento), meningite; 13 (68 por cento) ocorreram em crianças menores de 2 anos e 16 (84 por cento) em menores de 5 anos. A letalidade foi de 10 por cento. Dos 13 casos em menores de 2 anos, sete (54 por cento) foram SpNSPn e 10 (77 por cento) foram causados por sorotipos contemplados na vacina antipneumocócica conjugada 7-valente. Foram isolados 10 sorotipos, sendo mais freqüentes o 14, 6B e 23F, todos sensíveis à ceftriaxona. Dos três casos de meningite, dois foram causados por SpNSPn. CONCLUSÃO: A maioria das DPI ocorreu em menores de 2 anos de idade; 77 por cento das cepas e 86 por cento dos sorotipos de SpNSPn estão contemplados pela vacina antipneumocócica conjugada 7-valente.

OBJECTIVE: Invasive pneumococcal disease (IPD) primarily affects children less than 5 years old, the elderly and certain at-risk groups; especially people infected by the human immunodeficiency virus (HIV). The objective of this study was to analyze invasive pneumococcal diseases (IPD) in children and adolescents infected by the human immunodeficiency virus (HIV), with relation to morbidity, the case fatality ratio, pneumococcus serotypes, susceptibility to penicillin and ceftriaxone and to the proportion of susceptible and resistant Streptococcus pneumoniae (Sp) included in the 7-valent pneumococcal conjugate vaccine that has already been licensed. METHODS: A total of 19 cases of IPD were identified among HIV seropositive patients aged from 1 month to 20 years and hospitalized between 1993 and 2000. Data were recorded on standardized charts containing information on age, clinical diagnosis and progression, serotypes and the susceptibility to penicillin and ceftriaxone of the Sp strains identified in cultures. When the minimum inhibitory concentration was < 0.1 mcg/mL, Sp were defined as susceptible to penicillin (SpSPn), and all other strains were defined as not susceptible (SpNSPn). RESULTS: Of the 19 HIV seropositive cases with IPD, 16 (84 percent) had pneumonia and three (16 percent), had meningitis; 13 (68 percent) cases were children less than 2 years old and 16 (84 percent) were less than 5 years old. The case fatality ratio was 10 percent. Seven (54 percent) of the 13 cases less than 2 years old were SpNSPn and 10 (77 percent) were caused by serotypes covered by the 7-valent pneumococcal conjugate vaccine. From the 10 isolated serotypes the most frequent were 14, 6B and 23F, all them susceptible to ceftriaxone. From the three patients with meningitis, two were caused by SpNSPn. CONCLUSION: In this study most of the IPD occurred in children less than 2 years old; 77 percent of the strains and 86 percent of the serotypes of SpNSPn...

Fatores de risco clássicos e não clássicos para infecções pneumocócicas / Risk factors classic and non classic for pneumococcal infections

O Streptococcus pneumoniae é um patógeno que coloniza o trato respiratório superior, podendo-se disseminar causando doença pneumocócica invasiva (DIP) ou não invasiva. Dos fatores de risco conhecidos para as doenças invasivas a idade é dos mais importantes. Crianças com menos de 24 meses de vida têm altíssima incidência de DIP. Dos outros fatores de risco clássicos e não clássicos os mais significantes podem ser resumidos em: freqüência a creches e instituições correlatas, imunossupressão, especialmente ocasionada pelo HIV e pela asplenia funcional ou anatômica, cardiovasculopatias e nefropatias crônicas, hepatopatias crônicas, doenças crônicas com acometimento pulmonar, incluindo asma, fumo e cigarro, diabetes mellitus, fístulas liquóricas, infecções virais respiratórias prévias, variações de raças e populacionais e gênero masculino. Vários desses fatores mencionados contribuem também para o estabelecimento do estado de portador em nasofaringe. É fundamental que crianças, principalmente com fatores de risco importantes, recebam vacina conjugada antipneumocócica já nos primeiros meses de vida, medida preventiva mais eficaz contra o patógeno.

Identification of males at increased risk for genital human papillomavirus (HPV) infection among patients referred for urological consultation.

In order to identify the probable risk groups, this study analysed the reasons for referral to a urological clinic and the indications for peniscopy among males with suspected genital HPV infection. The patients were derived from a series of 2370 consecutive males attending our urological clinic in Sao Paulo, of whom 652 males (27.5%) aroused a clinical suspicion of genital HPV infection. Patients were categorized into 7 groups, according to their indications for referral. Altogether, 459 (70.4%) were examined by peniscopy, and 403 (87.8%) were biopsied and tested for HPV by Hybrid Capture 2 (HCII) for both oncogenic and non-oncogenic HPV types. The most frequent indication for referral in HPV-positive males (both in peniscopy and by HCII) was other diseases (group 6), followed by those who had partners HPV positive (group 1). The highest HPV prevalence (42%) was found among those with visible lesions, followed by those (38%) with previous condyloma. Of the other STDs, urethritis was significantly associated with HPV detection. Importantly, HPV was detected in 27% of the males referred due to other conditions non-related to HPV. We conclude that the risk groups for genital HPV infections include males with visible lesions, previous history of condyloma, those with HPV-positive partners, recurrent balanitis, as well as those with other STDs. Urethritis is significantly associated with HPV. The most frequent indications for referral to urological clinic among HPV-positive males were other, non-HPV-related conditions. The HPV prevalence rate of 27% among these patients warrants them to be considered as another risk group for genital HPV infections.

Transtorno factício: um desafio para as diversas especialidades / Factitious disease: a challenge to several specialities

O transtorno factício caracteriza-se por produção de sintomas por parte do paciente de forma intencional, com o intuito de assumir o papel de paciente. É uma doença de gravesimplicações clínicas e que gera elevados gastos para o sistema de saúde público. O caso da paciente DLK, ilustrado nesse relato, exemplifica bem esse transtorno. A paciente iniciou suas queixas clínicas com alucinações visuais e um quadro delirante. Com o tempo surgiram queixasde dores abdominais, retenção urinária, fraqueza muscular e um quadro psicótico de difícil resolução...

In factitious disease there is an intentional production of symptoms by the patient, with the intent of being sick in front of others. It is a disease with serious clinical implications and these patients represent a enormous waste of money to public health. The case of DLKexemplifies it. The complaints of the patient began with delusions and visual hallucinations. She developed abdominal pains, urinary retention, weakness and psychotic symptoms of difficultmanagement. She had gone to several hospitals, submitted to many exams and a laparoscopy, all with frustrating results. There was a progressive and important improvement by theestablishment of a positive relation with the patient and reduction of her medications...