Dengue virus 3 genotype I (GI) lineage 1 (L1) isolates elicit differential cytopathic effect with syncytium formation in human glioblastoma cells (U251).

BACKGROUND: Dengue virus (DENV) is a Flaviviridae member classified into four antigenically distinct serotypes (DENV 1, 2, 3, and 4) and further subdivided genotypes. DENV3 is subdivided into four or five genotypes, depending on the classification adopted. Despite their high genetic proximity, as revealed by phylogenetic complete polyprotein analysis, DENV3 MG-20 and DENV3 PV_BR showed different neurovirulence in mice models. Our group identified six amino acid mutations in protein E, including the E62K and E123Q, which may affect interactions of hydrophobic clusters on domain II, thus leading to the observed differences in the studied viruses. METHODS: Human glioblastoma cells (U251) derived from a malignant glioblastoma tumor by explant technique were infected by the DENV3 GIL1 isolates DENV3 MG-20 and DENV3 PV_BR and analyzed by plaque assays and titration, optical, immunofluorescence, and transmission electronic microscopy. RESULTS: The two isolates showed different cytopathic effects (CPE) and fusogenic patterns, further confirmed by indirect immunofluorescence. Transmission electron microscopy revealed intense cytopathic effects in DENV3 MG-20 infected U251 cells, displaying endoplasmic reticulum hypertrophy and turgid vesicles with proteins and multiple viruses, distinct from DENV3 PV_BR infected cells. It is hypothesized that the different amino acids in the DENV3 MG-20 isolate are related to an increased membrane fusion ability in viral infection, thus facilitating immune system evasion and increased chances of central nervous system cell infection. CONCLUSION: These results emphasize the biological differences between the isolates, which could be a critical factor in host-virus interaction and severe dengue development. Our study presents comparative results of highly similar isolates with the potential to generate more subsidies for a deeper understanding of the DENV pathogenesis. The neurotropism of the isolate DENV3 MG-20 (belonging to the DENV3 GI L1 genotype) showing infection of nervous system cells (U251) could contribute to understanding neurological dengue disease.

Dengue virus 3 genotype I shows natural changes in heparan sulphate binding sites, cell interactions, and neurovirulence in a mouse model.

Dengue virus (DENV) is the most prevalent pathogen of the Flaviviridae family. Due to the considerable increase in DENV incidence and spread, symptoms such as CNS involvement have increased. Heparan sulphate (HS) was the first molecule identified as an adhesion factor for DENV in mammalian cells. Viral phenotypes with different HS interactions are associated with various clinical symptoms, including neurological alterations. Here, using in silico analyses, in vitro studies, and the in vivo mouse model, we characterized two natural circulating DENV3 genotype I (GI) lineage 1 (L1) in Brazil-DENV3 MG-20 (from Minas Gerais) and DENV3 PV_BR (from Rondônia) that present divergent neurovirulent profiles and sensitivity to sulphated molecules. We identified substitutions at the viral envelope (E) in positions 62 and 123 as likely responsible for the differences in neurovirulence. The E62K and E123Q substitutions in DENV3 MG-20 and DENV3 PV_BR, respectively, greatly influenced in silico electrostatic density and heparin docking results. In vivo, mice inoculated with DENV3 MG-20 died, but not those infected with DENV3 PV_BR. The clinical symptoms, such as paralysis of the lower limbs and meningoencephalitis, and histopathology, also differed between the inoculated groups. In vitro heparin and heparinases assays further demonstrated the biological impact of these substitutions. Other characteristics that have been previously associated with alterations in cell tropism and neurovirulence, such as changes in the size of lysis plaques and differences in cytopathic effects in glioblastoma cells, were also observed.

Resting Tremor after Mild Head Injury: Case Report

Over the past few decades, it has been recognized that traumatic brain injury (TBI) may result in various movement disorders. However, moderate or mild TBI only rarely causes persistent post-traumatic movement disorders. In the present report, we describe a case of secondary tremor due to amild head injury with a transitory loss of consciousness. A 26- year-old man developed an isolated rest tremor of the hands and legs without other neurologic signs. The interval between the head trauma and the onset of the symptomswas 4 months. Neuroimaging studies reveled gliosis in the lentiform nucleus. Haloperidol administration resulted in tremor reduction. A rest tremor, similar to essential tremor, can be a rare complication of head trauma. Haloperidolmay be an effective and safe treatment modality for post-traumatic tremor. Further studies are needed to clarify the optimal drug for the treatment of post-traumatic tremor.

As dimensões da satisfação dos usuários do Programa Saúde da Família: confiabilidade e empatia / The Dimensions of user satisfaction of the Family Health Program: trust and empathy / Las dimensiones de la satisfacción de los usuarios del Programa Salud de la Familia: confiabilidad y empatía

OBJETIVO: Descrever a percepção dos usuários das equipes do Programa Saúde da Família (PSF) da zona urbana de Montes Claros, Minas Gerais, sobre as dimensões da satisfação relacionadas à Confiabilidade e à Empatia; analisar o perfil sociodemográfico da população entrevistada frente à Confiabilidade e à Empatia. MÉTODOS: Pesquisa quantitativa; descritiva por meio do SERVQUAL. Foram entrevistados 319 usuários e a análise foi realizada por meio da estatística descritiva e as associações estatisticamente significantes. RESULTADOS: A maioria dos entrevistados é do sexo feminino (80,6 por cento), encontra-se na faixa etária de 18 a 30 anos (38,2 por cento), possui ensino médio completo (32,3 por cento), renda familiar de 1 a 3 salários mínimos (76,8 por cento), reside nos bairros há mais de 10 anos (52,6 por cento) e tem boa percepção dos serviços prestados quanto à Confiabilidade e à Empatia. CONCLUSÃO: Foi possível descrever os aspectos que envolvem a Confiabilidade e a Empatia dos usuários do PSF, que merecem ser considerados na reestruturação dos processos de atenção à saúde da população.

OBJECTIVE: Describe users' perceptions of teams from the Family Health Program (PSF) of the urban area of Montes Claros, Minas Gerais, regarding the dimensions of satisfaction related to trust and empathy; analyze the demographic profile of the population interviewed in relationship to their perceptions of trust and empathy. METHODS: Quantitative descriptive survey; using SERVQUAL. 319 users were interviewed and the analysis was performed using descriptive statistics to identify statistically significant associations. RESULTS: Most respondents were: female (80.6 percent), ages 18 to 30 years (38.2 percent), had completed high school (32.3 percent), had a family income of 1 to 3 times the minimum wage (76.8 percent), lived in the district for over 10 years (52.6 percent), and had good understanding of services as related to trust and empathy. CONCLUSION: We describe the aspects involving trust and empathy for users of the PSF, which must be considered when restructuring the processes of health care for the population.

OBJETIVO: Describir la percepción de los usuarios de los equipos del Programa Salud de la Familia (PSF) de la zona urbana de Montes Claros, Minas Gerais, sobre las dimensiones de la satisfacción relacionadas a la Confiabilidad y la Empatia; analizar el perfil sociodemográfico de la población entrevistada frente a la Confiabilidad y a la Empatía. MÉTODOS: Investigación cuantitativa; descriptiva por medio del SERVQUAL. Fueron entrevistados 319 usuarios y el análisis fue realizado por meio da estadística descriptiva y las asociaciones estadísticamente significativas. RESULTADOS: La mayoría de los entrevistados es del sexo feminino (80,6 por ciento), se encuentra en el grupo etáreo de 18 a 30 años (38,2 por ciento), posee secundaria completa (32,3 por ciento), ingreso familiar de 1 a 3 salarios mínimos (76,8 por ciento), reside en los barrios hace más de 10 años (52,6 por ciento) y tiene buena percepción de los servicios prestados en cuanto a la Confiabilidad y a la Empatía. CONCLUSIÓN: Fue posible describir los aspectos que involucran a la Confiabilidad y la Empatía de los usuarios del PSF, que merecem ser considerados en la reestructuración de los procesos de atención a la salud de la población.