Papers by José Antonio Diniz
Brazilian Journal of Health Review
Introdução: Os arbovírus neurotrópicos representam um importante problema de saúde em escala glo... more Introdução: Os arbovírus neurotrópicos representam um importante problema de saúde em escala global, sendo necessário a utilização de medidas de prevenção, controle e diagnóstico. Objetivo: Avaliar a efetividade da infecção de dois arbovírus neurotrópicos, vírus do Nilo Ocidental (WNV) e vírus da Encefalite de Saint Louis (SLEV), em células de neuroblastoma humano (IMR-32). Metodologia: As suspensões-estoques dos vírus foram tituladas por ensaio de placa com incubação de 120 h para WNV e 168 h para SLEV. Os isolados foram inoculados em garrafas para cultura de células que apresentavam 80% de confluência da monocamada celular e observadas por microscopia de contraste de fase por 72 h pós infecção a fim de verificar alterações morfológicas sugestivas de efeito citopático (ECP). Em seguida, as culturas foram submetidas ao ensaio de Imunofluorescência Indireta (IFI) para detecção de antígenos virais. Resultados: O título infeccioso do SLEV apresentou-se mais baixo em relação ao WNV. E...
Frontiers in Cellular and Infection Microbiology, 2022
Ocular infection with Toxoplasma gondii causes toxoplasmosis in mice. However, following ocular i... more Ocular infection with Toxoplasma gondii causes toxoplasmosis in mice. However, following ocular infection with tachyzoites, the cause of the accompanying progressive changes in hippocampal-dependent tasks, and their relationship with the morphology and number of microglia, is less well understood. Here, in 6-month-old, female BALB/c mice, 5 μl of a suspension containing 48.5 × 106 tachyzoites/ml was introduced into the conjunctival sac; control received an equal volume of saline. Before and after instillation, all mice were subject to an olfactory discrimination (OD) test, using predator (cat) feces, and to an open-field (OF) task. After the behavioral tests, the animals were culled at either 22 or 44 days post-instillation (dpi), and the brains and retinas were dissected and processed for immunohistochemistry. The total number of Iba-1-immunolabeled microglia in the molecular layer of the dentate gyrus was estimated, and three-dimensional reconstructions of the cells were evaluated...
All neurological diseases have their pathological progress influenced by microglial cells, which ... more All neurological diseases have their pathological progress influenced by microglial cells, which react to insults with multiple morphofunctional phenotypes. However, the complete morphological spectrum of the reactive microglia, revealed by the three-dimensional microscopic reconstruction, has not been explored in detail in virus limbic encephalitis. Here, we used anatomical series of brain sections from a previous study using Piry arbovirus encephalitis, expanding the original investigation to CA1/CA2. The morphological response of the homeostatic and reactive microglia was investigated 8 days after infection. The hierarchical cluster and linear discriminant function analysis of multimodal morphometric features allowed to distinguish between the microglial morphology of infected individuals and controls. In order to represent more broadly the morphological spectrum of the microglia of each cluster, representative cells of the homeostatic and reactive microglia were chosen, using th...
Frontiers in Neuroanatomy, 2016
Revista Pan-Amazônica de Saúde, 2015
Santos DVV, et al. Detecção de atividade de fosfatase ácida nos hemócitos de Biomphalaria glabrat... more Santos DVV, et al. Detecção de atividade de fosfatase ácida nos hemócitos de Biomphalaria glabrata 49 CONCLUSÃO Com este trabalho demonstra-se a presença de grandes lisossomos em hemócitos granulócitos de caramujo da espécie B. glabrata, pela identificação de compartimentos ácidos contendo a enzima fosfatase ácida. Estas organelas apresentaram grânulos envoltos por membrana com formas e disposições variadas por todo o citoplasma celular. Detection of acid phosphatase activity in hemocytes of Biomphalaria glabrata
Revista Pan-Amazônica de Saúde, 2011
In this study, we analyzed a fecal sample of a female infant with paralysis and other clinical sy... more In this study, we analyzed a fecal sample of a female infant with paralysis and other clinical symptoms that resembled poliomyelitis. Negative staining electron microscopy showed viral particles with a diameter of approximately 120 nm and displaying a crown-like appearance with surface projections. Ultrathin sections showed particles budding from the membranes of the Golgi apparatus. Based on these results, we propose the association of this virus with the neurological disorder and tentatively assign it to the Coronaviridae family. Further studies are required on this proposed relationship.
PLoS ONE, 2011
An enriched environment has previously been described as enhancing natural killer cell activity o... more An enriched environment has previously been described as enhancing natural killer cell activity of recognizing and killing virally infected cells. However, the effects of environmental enrichment on behavioral changes in relation to virus clearance and the neuropathology of encephalitis have not been studied in detail. We tested the hypothesis that environmental enrichment leads to less CNS neuroinvasion and/or more rapid viral clearance in association with T cells without neuronal damage. Stereology-based estimates of activated microglia perineuronal nets and neurons in CA3 were correlated with behavioral changes in the Piry rhabdovirus model of encephalitis in the albino Swiss mouse. Two-month-old female mice maintained in impoverished (IE) or enriched environments (EE) for 3 months were behaviorally tested. After the tests, an equal volume of Piry virus (IEPy, EEPy)-infected or normal brain homogenates were nasally instilled. Eight days postinstillation (dpi), when behavioral changes became apparent, brains were fixed and processed to detect viral antigens, activated microglia, perineuronal nets, and T lymphocytes by immuno-or histochemical reactions. At 20 or 40 dpi, the remaining animals were behaviorally tested and processed for the same markers. In IEPy mice, burrowing activity decreased and recovered earlier (8-10 dpi) than open field (20-40 dpi) but remained unaltered in the EEPy group. EEPy mice presented higher T-cell infiltration, less CNS cell infection by the virus and/or faster virus clearance, less microgliosis, and less damage to the extracellular matrix than IEPy. In both EEPy and IEPy animals, CA3 neuronal number remained unaltered. The results suggest that an enriched environment promotes a more effective immune response to clear CNS virus and not at the cost of CNS damage.
Memórias do Instituto Oswaldo Cruz, 2002
The Mojuí dos Campos virus (MDCV) was isolated from the blood of an unidentified bat (Chiroptera)... more The Mojuí dos Campos virus (MDCV) was isolated from the blood of an unidentified bat (Chiroptera) captured in Mojuí dos Campos, Santarém, State of Pará, Brazil, in 1975 and considerated to be antigenically different from other 102 arboviruses belonging to several antigenic groups isolated in the Amazon region or another region by complement fixation tests. The objective of this work was to develop a morphologic, an antigenic and physicochemical characterization of this virus. MDCV produces cytopathic effect in Vero cells, 24 h post-infection (p.i), and the degree of cellular destruction increases after a few hours. Negative staining electron microscopy of the supernatant of Vero cell cultures showed the presence of coated viral particles with a diameter of around 98 nm. Ultrathin sections of Vero cells, and brain and liver of newborn mice infected with MDCV showed an assembly of the viral particles into the Golgi vesicles. The synthesis kinetics of the proteins for MDCV were similar to that observed for other bunyaviruses, and viral proteins could be detected as early as 6 h p.i. Our results reinforce the original studies which had classified MDCV in the family Bunyaviridae, genus Bunyavirus as an ungrouped virus, and it may represent the prototype of a new serogroup.
The American Journal of Tropical Medicine and Hygiene, 2005
This paper reports the results of serologic, structural, biochemical, and genetic studies indicat... more This paper reports the results of serologic, structural, biochemical, and genetic studies indicating that Araguari virus, a previously unassigned viral agent, is a member of the family Orthomyxoviridae and genus Thogotovirus. Araguari virus has six RNA fragments; biologically, it shares several properties with other viruses in the family Orthomyxoviridae. Nucleotide sequencing of the RNA segments 4 (glycoprotein) and 5 (nucleoprotein) of Araguari virus aligned with the orthomyxoviruses, showing the closest relationship with Thogoto virus (sequence similarity ס 61.9% and 69.1%, respectively, for glycoprotein and nucleoprotein), but also sharing a more distant similarity with Dhori and Influenza C viruses, especially for the glycoprotein gene. Based on these results, we propose that Araguari virus should be assigned as a new member of the family Orthomyxoviridae and genus Thogotovirus.
The American Journal of Tropical Medicine and Hygiene, 2006
Primary cultures of embryonic murine neurons and newborn mouse astrocytes were inoculated with We... more Primary cultures of embryonic murine neurons and newborn mouse astrocytes were inoculated with West Nile virus (WNV) strain NY385-99 to compare the pathogenesis of WNV infection in these types of CNS cells. Two different outcomes were observed. WNV infection in the neurons was rapidly progressive and destructive; within 5 days, all of the neurons were destroyed through apoptosis. WNV infection in the astrocytes evolved more slowly and did not seem to be highly lethal to the cells. The infected astrocytes continued to produce infectious virus (10 4.6 -10 6.5 PFU/mL) for 114 days, in a permissive, persistent infection. During this period, WNV antigen could be shown in the cytoplasm of the infected astrocytes by immunocytochemical assay, transmission electron microscopy of ultrathin sections, and in the cell culture medium by complement fixation test. Our results with this in vitro experimental murine cell model indicate that astrocytes can develop chronic or persistent infection with WNV, suggesting that these cells may play a role in the maintenance of WNV in the CNS.
European Journal of Inflammation, 2013
We previously demonstrated in young mice that in comparison with animals raised in an impoverishe... more We previously demonstrated in young mice that in comparison with animals raised in an impoverished environment (IE), animals from an enriched environment (EE) show more severe dengue disease, associated with an increased expansion of memory T target cells. Because active older adults show less functional decline in T-cell adaptive immunity, we hypothesized that aged mice from EE would show higher mortality and T-lymphocyte expansion than mice from IE. To test this hypothesis, we administered serial i.p. injections of anti-DENV2 hyperimmune serum, followed 24 h later by DENV3 (genotype III)-infected brain homogenate. Control mice received equal volumes of serum but received uninfected brain homogenate. The presence of virus or viral antigens was indirectly detected by real-time quantitative RT-PCR and immunohistochemistry. Compared to infected IE animals, EE mice, independent of age, showed higher mortality and more intense clinical signs. Compared to young mice, the higher mortality...
Parasitology Research, 2002
Genome Announcements, 2014
We report here the first complete open reading frame (ORF) genome sequence of Xiburema virus (XIB... more We report here the first complete open reading frame (ORF) genome sequence of Xiburema virus (XIBV), that of strain BE AR362159, isolated from mosquitoes ( Sabethes intermedius ) in Sena Madureira, Acre state, northern Brazil. All genes showed similarities with those belonging to members of the family Rhabdoviridae .
Genome Announcements, 2014
We report here the first nearly complete genome sequence related to curionopolis virus (CURV), th... more We report here the first nearly complete genome sequence related to curionopolis virus (CURV), that of strain AR440009, isolated from a pool of Culicoides sp. midges in Serra Norte, Pará State, northern Brazil. All genes showed similarities to those belonging to members of the family Rhabdoviridae .
Acta Cirurgica Brasileira, 2000
Em virtude da extensa utilização empírica do óleo de copaíba em tratamentos ginecológicos, este t... more Em virtude da extensa utilização empírica do óleo de copaíba em tratamentos ginecológicos, este trabalho visa realizar um estudo ultraestrutural do colo uterino de ratas após aplicação deste óleo. Foram utilizadas 12 ratas adultas distribuídas em 4 grupos: Água, Milho, Padrão e Copaíba. Os animais foram submetidos a ooforectomia bilateral e após 21 dias foi iniciada a aplicação das substâncias. A eutanásia foi realizada em datas pré-detreminadas (7º, 14º e 21º dias). Os grupos Padrão, Água e Milho apresentaram epitélio com duas a três fileiras de células poliédricas e núcleos de diferentes tamanhos e formas. Já no grupo copaíba, o epitélio era constituído de várias fileiras de células (8 a 10 fileiras). Concluiu-se que o óleo de copaíba promoveu aumento de fileira de células da camada epitelial com queratinização no colo uterino de ratas ooforectomizadas.
Because an enriched environment (EE) enhances T-cell activity and T-lymphocytes contribute to imm... more Because an enriched environment (EE) enhances T-cell activity and T-lymphocytes contribute to immunopatho genesis during heterologous dengue virus (DENV) infections, we hypothesised that an EE increases dengue severity. To compare single serotype (SS) and antibody-enhanced disease (AED) infections regimens, serial intraperitoneal were performed with DENV3 (genotype III) infected brain homogenate or anti-DENV2 hyperimmune serum followed 24 h later by DENV3 (genotype III) infected brain homogenate. Compared AED for which significant differences were detected between the EE and impoverished environmental (IE) groups (Kaplan-Meyer log-rank test, p = 0.0025), no significant differences were detected between the SS experimental groups (Kaplan-Meyer log-rank test, p = 0.089). Survival curves from EE and IE animals infected with the AED regimen were extended after corticoid injection and this effect was greater in the EE than in the IE group (Kaplan-Meyer log-rank test, p = 0.0162). Under t...
Revista do Instituto de Medicina Tropical de São Paulo, 2004
We report the isolation of Fonsecaea pedrosoi from thorns of the plant Mimosa pudica L. at the pl... more We report the isolation of Fonsecaea pedrosoi from thorns of the plant Mimosa pudica L. at the place of infection identified by one of our patients. Clinical diagnosis of chromoblastomycosis was established by direct microscopic examination and cultures from the patient's lesion. The same species was isolated from the patient and from the plant. Scanning electron microscopy of the surface of the thorns showed the characteristic conidial arrangement of F. pedrosoi. These data indicate that M. pudica could be a natural source of infection for the fungus F. pedrosoi.
Tissue and Cell, 2005
The fine structure and differential cell count of blood and coelomic exudate leukocytes were stud... more The fine structure and differential cell count of blood and coelomic exudate leukocytes were studied with the aim to identify granulocytes from Ameiva ameiva, a lizard distributed in the tropical regions of the Americas. Blood leukocytes were separated with a Percoll cushion and coelomic exudate cells were obtained 24 h after intracoelomic thioglycollate injection. In the blood, erythrocytes, monocytes, thrombocytes, lymphocytes, plasma cells and four types of granulocytes were identified based on their morphology and cytochemistry. Types I and III granulocytes had round intracytoplasmic granules with the same basic morphology; however, type III granulocyte had a bilobued nucleus and higher amounts of heterochromatin suggesting an advance stage of maturation. Type II granulocytes had fusiformic granules and more mitochondria. Type IV granulocytes were classified as the basophil mammalian counterpart based on their morphology and relative number. Macrophages and granulocytes type III were found in the normal coelomic cavity. However, after the thioglycollate injection the number of type III granulocyte increased. Granulocytes found in the coelomic cavity were related to type III blood granulocyte based on the morphology and cytochemical localization of alkaline phosphatase and basic proteins in their intracytoplasmic granules. Differential blood leukocyte counts showed a predominance of type III granulocyte followed by lymphocyte, type I granulocyte, type II granulocyte, monocyte and type IV granulocyte. Taken together, these results indicate that types I and III granulocytes correspond to the mammalian neutrophils/heterophils and type II to the eosinophil granulocytes.
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Papers by José Antonio Diniz