Testicular damage without clinical manifestations in BALB/c mice experimentally infected with Zika virus.

In 2015-2016, the Zika virus (ZIKV) caused a major epidemic in the Americas, increasing cases of microcephaly and Guillain-Barré syndrome. During this period, the discovery of ZIKV sexual transmission intensified studies on the impact of this virus on the reproductive organs. For this study, 2-month-old male BALB/c mice were infected with 1.26 x 106 PFU/mL of ZIKV in solution via the intravenous route. After three, seven, and fourteen days post-infection (DPI), blood and testicle samples were obtained to detect ZIKV RNA. The authors observed that the infected animals had slower weight gain than the control group. Viremia occurred only at 3DPI, and the ZIKV RNA was detected in one testis sample at 7DPI. The histopathological analysis of this organ revealed intense disorganization of the seminiferous tubules' structure, inflammatory infiltrate, necrosis, hemorrhage, fluid accumulation, congestion of blood vessels, and reduced sperm count. Ultrastructural analysis showed nuclear changes in tubule cells, activation of interstitial cells, and morphological changes in spermatozoa, in addition to fragmentation and decreased electron density of the genetic material of these cells. Thus, despite causing predominantly asymptomatic infections, ZIKV can cause significant subclinical and transient damage, including to male reproductive organs.

Influence of a Physiologically Formed Blood Clot on Pre-Osteoblastic Cells Grown on a BMP-7-Coated Nanoporous Titanium Surface.

Titanium (Ti) nanotopography modulates the osteogenic response to exogenous bone morphogenetic protein 7 (BMP-7) in vitro, supporting enhanced alkaline phosphatase mRNA expression and activity, as well as higher osteopontin (OPN) mRNA and protein levels. As the biological effects of OPN protein are modulated by its proteolytic cleavage by serum proteases, this in vitro study evaluated the effects on osteogenic cells in the presence of a physiological blood clot previously formed on a BMP-7-coated nanostructured Ti surface obtained by chemical etching (Nano-Ti). Pre-osteoblastic MC3T3-E1 cells were cultured during 5 days on recombinant mouse (rm) BMP-7-coated Nano-Ti after it was implanted in adult female C57BI/6 mouse dorsal dermal tissue for 18 h. Nano-Ti without blood clot or with blood clot at time 0 were used as the controls. The presence of blood clots tended to inhibit the expression of key osteoblast markers, except for Opn, and rmBMP-7 functionalization resulted in a tendency towards relatively greater osteoblastic differentiation, which was corroborated by runt-related transcription factor 2 (RUNX2) amounts. Undetectable levels of OPN and phosphorylated suppressor of mothers against decapentaplegic (SMAD) 1/5/9 were noted in these groups, and the cleaved form of OPN was only detected in the blood clot immediately prior to cell plating. In conclusion, the strategy to mimic in vitro the initial interfacial in vivo events by forming a blood clot on a Ti nanoporous surface resulted in the inhibition of pre-osteoblastic differentiation, which was minimally reverted with an rmBMP-7 coating.

Detection and Molecular Characterization of Enteric Viruses in Bivalve Mollusks Collected in Arraial do Cabo, Rio de Janeiro, Brazil.

Viral bivalve contamination is a recognized food safety hazard. Therefore, this study investigated the detection rates, seasonality, quantification, and genetic diversity of enteric viruses in bivalve samples (mussels and oysters). We collected 97 shellfish samples between March 2018 and February 2020. The screening of samples by qPCR or RT-qPCR revealed the detection of norovirus (42.3%), rotavirus A (RVA; 16.5%), human adenovirus (HAdV; 24.7%), and human bocavirus (HBoV; 13.4%). There was no detection of hepatitis A virus. In total, 58.8% of shellfish samples tested positive for one or more viruses, with 42.1% of positive samples contaminated with two or more viruses. Norovirus showed the highest median viral load (3.3 × 106 GC/g), followed by HAdV (median of 3.5 × 104 GC/g), RVA (median of 1.5 × 103 GC/g), and HBoV (median of 1.3 × 103 GC/g). Phylogenetic analysis revealed that norovirus strains belonged to genotype GII.12[P16], RVA to genotype I2, HAdV to types -C2, -C5, and -F40, and HBoV to genotypes -1 and -2. Our results demonstrate the viral contamination of bivalves, emphasizing the need for virological monitoring programs to ensure the quality and safety of shellfish for human consumption and as a valuable surveillance tool to monitor emerging viruses and novel variants.

Effects of rmBMP-7 on Osteoblastic Cells Grown on a Nanostructured Titanium Surface.

This study evaluates the effects of the availability of exogenous BMP-7 on osteoblastic cells' differentiation on a nanotextured Ti surface obtained by chemical etching (Nano-Ti). The MC3T3-E1 and UMR-106 osteoblastic cell lines were cultured for 5 and 7 days, respectively, on a Nano-Ti surface and on a control surface (Control-Ti) in an osteogenic medium supplemented with either 40 or 200 ng/mL recombinant mouse (rm) BMP-7. The results showed that MC3T3-E1 cells exhibited distinct responsiveness when exposed to each of the two rmBMP-7 concentrations, irrespective of the surface. Even with 40 ng/mL rmBMP-7, important osteogenic effects were noticed for Control-Ti in terms of cell proliferation potential; Runx2, Osx, Alp, Bsp, Opn, and Smad1 mRNA expression; and in situ ALP activity. For Nano-Ti, the effects were limited to higher Alp, Bsp, and Opn mRNA expression and in situ ALP activity. On both surfaces, the osteogenic potential of UMR-106 cultures remained unaltered with 40 ng/mL rmBMP-7, but it was significantly reduced when the cultures were exposed to the 200 ng/mL concentration. The availability of rmBMP-7 to pre-osteoblastic cells at the concentrations used alters the expression profile of osteoblast markers, indicative of the acquisition of a more advanced stage of osteoblastic differentiation. This occurs less pronouncedly on the nanotextured Ti and without reflecting in higher mineralized matrix production by differentiated osteoblasts on both surfaces.

Morphological Aspects and Viremia Analysis of BALB/c Murine Model Experimentally Infected with Dengue Virus Serotype 4.

Ever since its brief introduction in the Brazilian territory in 1981, dengue virus serotype 4 (DENV-4) remained absent from the national epidemiological scenario for almost 25 years. The emergence of DENV-4 in 2010 resulted in epidemics in most Brazilian states. DENV-4, however, remains one of the least studied among the four DENV serotypes. Despite being known as a mild serotype, DENV-4 is associated with severe cases and deaths and deserves to be investigated; however, the lack of suitable experimental animal models is a limiting factor for pathogenesis studies. Here, we aimed to investigate the susceptibility and potential tropism of DENV-4 for liver, lung and heart of an immunocompetent mice model, and to evaluate and investigate the resulting morphological and ultrastructural alterations upon viral infection. BALB/c mice were inoculated intravenously with non-neuroadapted doses of DENV-4 isolated from a human case. The histopathological analysis of liver revealed typical alterations of DENV, such as microsteatosis, edema and vascular congestion, while in lung, widespread areas of hemorrhage and interstitial pneumonia were observed. While milder alterations were present in heart, characterized by limited hemorrhage and discrete presence of inflammatory infiltrate, the disorganization of the structure of the intercalated disc is of particular interest. DENV-4 RNA was detected in liver, lung, heart and serum of BALB/c mice through qRT-PCR, while the NS3 viral protein was observed in all of the aforementioned organs through immunohistochemistry. These findings indicate the susceptibility of the model to the serotype and further reinforce the usefulness of BALB/c mice in studying the many alterations caused by DENV.

Comparative analysis of liver involvement caused by two DENV-2 lineages using an immunocompetent murine model.

Dengue (DEN) is the most prevalent arbovirus among humans, and four billion people live at risk of infection. The clinical manifestations of DEN are variable, and the disease may present subclinically or asymptomatically. A quarter of patients develop classical dengue (CD) or severe dengue (SD), which is potentially lethal and involves vascular permeability changes, severe hemorrhage and organ damage. The involvement of the liver is a fairly common feature in DEN, and alterations range from asymptomatic elevation of transaminases to acute liver failure. Since its introduction in Brazil in 1990, two strains of Dengue virus (DENV) serotype 2 (DENV-2) have been detected: Lineage I, which is responsible for an outbreak in 1991, and Lineage II, which caused an epidemic greater than the previous one and had a different epidemiological profile. To date, studies on different strains of the same serotype/genotype and their association with disease severity are scarce. In addition, one of the greatest challenges regarding the study of DEN pathogenesis and the development of drug and vaccine therapies is the absence of an animal model that reproduces the disease as it occurs in humans. The main goals of this study were to assess BALB/c mouse susceptibility experimentally infected by two distinct DENV-2 strains and characterize possible differences in the clinical signs and alterations induced in the liver resulting from those infections. Mice infected by the two DENV-2 lineages gained less weight than uninfected mice; however, their livers were slightly heavier. Increased AST and AST levels were observed in infected mice, and the number of platelets increased in the first 72 h of infection and subsequently decreased. Mice infected with both lineages presented leukocytosis but at different times of infection. The histopathological changes induced by both lineages were similar and comparable to the changes observed in DEN fatal cases. The viral genome was detected in two liver samples. The results demonstrate the susceptibility of BALB/c mice to both DENV-2 lineages and suggest that the changes induced by those strains are similar, although for some parameters, they are manifested at different times of infection.

Distribución espacial del seguimiento de cáncer de mama en un área rural de la estrategia de salud familiar / Spatial distribution of breast cancer tracking in a rural area of the family health strategy / Distribuição espacial do rastreamento do câncer de mama em uma área rural da estratégia de saúde da familia

OBJETIVO: Demostrar mediante georreferenciación micro áreas con debilidades en el cribado del cáncer de mama en el área de Family Strategy Caxirimbu, área rural de Caxias, Maranhão, Brasil. MÉTODOS: Investigación epidemiológica seccional. Se entrevistó a 211 mujeres, con edades entre 40 y 69 años, entre el 1 de abril y el 1 de septiembre de 2015. La técnica de georreferenciación se utilizó con la producción de coordenadas geográficas utilizando equipos GPS y la producción de mapas de distribución espacial. RESULTADOS: En la distribución espacial, se encontró que el cribado del cáncer de mama en el área de Caxirimbu tiene un mayor alcance en las micro áreas en los márgenes de MA 034 y en las micro áreas cercanas a la unidad de salud. En esta cobertura, se encontró que de las 211 mujeres entrevistadas, 133 (63.0%) ya se habían sometido a una mamografía, estando cerca del parámetro indicado por el Ministerio de Salud, que es al menos el 70.0%. 42.0% (n = 56) ya se había sometido a una mamografía; 36.0% (n = 48), dos o tres mamografías, y el 22.0% (n = 29) informó haber realizado más de cuatro mamografías. El mapa mostró que 11 lugares de este examen son inaccesibles para las mujeres. En la distribución espacial, hubo una mayor distribución para la mamografía irregular, convergiendo con su frecuencia del 80.0%. CONCLUSIÓN: La distribución espacial de los intervalos practicados por las mujeres para los exámenes de mamografía demostró la mayor parte de la ejecución y la ocurrencia de intervalos inadecuados, identificando la necesidad de implementar una detección organizada

OBJETIVO: Demonstrar por meio do georreferenciamento microáreas com fragilidades no rastreamento do câncer de mama em na área da estratégia de Família do Caxirimbu, zona rural de Caxias - Maranhão. MÉTODOS: Pesquisa epidemiológica do tipo seccional. Foram pesquisadas 211 mulheres de 40 a 69 anos entre 01 de abril a 01 de setembro de 2015. Utilizou-se a técnica do georreferenciamento com produção de coordenadas geográficas por meio de equipamento GPS e produção de mapas de distribuição espacial. RESULTADOS: Na distribuição espacial verificou-se que o rastreamento do câncer de mama na área de abrangência do Caxirimbu tem maior alcance nas microáreas as margens da MA 034 e em microáreas próxima a unidade de saúde. Nesta cobertura, verificou-se que das 211 mulheres entrevistadas, 133 (63,0%) já haviam realizado a mamografia ficando próximo do parâmetro indicado pelo ministério da saúde que é de no mínimo 70,0%. 42,0% (n=56) já haviam realizado 1 mamografia; 36,0% (n=48), 2 a 3 mamografias e; 22,0% (n=29) informou ter realizado mais de 4 mamografias. A execução da mamografia e sua regularidade teve uma frequência de 26,0% e 20,0%, respectivamente. O mapa mostrou que 11 localidades esse rastreamento está inacessível para as mulheres. Verificou-se na distribuição espacial maior distribuição para execução irregular da mamografia convergindo com sua frequência de 80,0%. CONCLUSÃO: A distribuição espacial dos intervalos praticados pelas mulheres para a realização dos exames de mamografia demonstrou que a maioria da execução e ocorrência de intervalos inadequados, identificando a necessidade de implementação de um rastreamento organizado

OBJECTIVE: To demonstrate by means of georeferencing micro-areas with weaknesses in screening of breast cancer in the area of ​​the Caxirimbu Family Strategy, rural zone of Caxias, Maranhão state, Brazil. METHODS: Sectional epidemiological research. A total of 211 women aged 40 to 69 years were surveyed between April 1 and September 1, 2015. The georeferencing technique was used with the production of geographical coordinates using GPS equipment and the production of spatial distribution maps. RESULTS: In the spatial distribution, it was found that breast cancer screening in the area covered by Caxirimbu has greater reach in the micro areas on the margins of MA 034 and in micro areas close to the health unit. In this coverage, it was found that of the 211 women interviewed, 133 (63.0%) had already undergone mammography, being close to the parameter indicated by the Ministry of Health, which is at least 70.0%. 42.0% (n= 56) had already undergone one mammogram; 36.0% (n= 48), two to three mammograms and; 22.0% (n= 29) reported having performed more than four mammograms. The map showed that 11 locations that tracking is inaccessible to women. In the spatial distribution, there was a greater distribution for irregular mammography performance, converging with its frequency of 80.0%. CONCLUSION: The spatial distribution of the intervals practiced by women for mammography exams demonstrated that most of the execution and occurrence of inadequate intervals, identifying the need to implement an organized screening

First detection of dengue virus in the saliva of immunocompetent murine model.

The lack of an experimental animal model for the study of dengue pathogenesis is a limiting factor for the development of vaccines and drugs. In previous studies, our group demonstrated the susceptibility of BALB/c mice to infection by dengue virus (DENV) 1 and 2, and the virus was successfully isolated in several organs. In this study, BALB/c mice were experimentally infected intravenously with DENV-4, and samples of their saliva were collected. Viral RNA extracted from the saliva samples was subjected to qRT-PCR, with a detection limit of 0.002 PFU/mL. The presence of DENV-4 viral RNA was detected in the saliva of two mice, presenting viral titers of 109 RNA/mL. The detection of DENV RNA via saliva sampling is not a common practice in dengue diagnosis, due to the lower detection rates in human patients. However, the results observed in this study seem to indicate that, as in humans, detection rates of DENV RNA in mouse saliva are also low, correlating the infection in both cases. This study reports the first DENV detection in the saliva of BALB/c immunocompetent mice experimentally infected with non-neuroadapted DENV-4.

Effects of botulinum toxin type A for spastic foot in post-stroke patients enrolled in a rehabilitation program.

UNLABELLED: The objective of this study was to evaluate the effects of botulinum toxin type A (BTX-A) on spastic foot in stroke patients in a rehabilitation program. METHOD: Hemiparetic stroke patients (n=21) enrolled in a rehabilitation program were divided into two groups. The first group (n=11) received a total of 300 UI BTX-A, and the second group (n=10) received 100 UI BTX-A. All patients were assessed at baseline and 2, 4, 8 and 12 weeks after injection for Modified Ashworth Score, time walking 10 meters, and the Functional Independence Measure (mFIM) motor score. RESULTS: The higher-dose group exhibited a significant improvement in spasticity, and both groups showed an improvement in time walking 10 meters and mFIM, with no significant differences between them. CONCLUSIONS: Our findings suggest that gains in gait velocity and functional independence were not correlated to BTX-A dose.

Effects of botulinum toxin type A for spastic foot in post-stroke patients enrolled in a rehabilitation program / Efeitos da toxina botulinica tipo A no pe espastico em pacientes pos-acidente vascular encefalico inseridos em programa de reabilitacao

The objective of this study was to evaluate the effects of botulinum toxin type A (BTX-A) on spastic foot in stroke patients in a rehabilitation program. Method: Hemiparetic stroke patients (n=21) enrolled in a rehabilitation program were divided into two groups. The first group (n=11) received a total of 300UI BTX-A, and the second group (n=10) received 100 UI BTX-A. All patients were assessed at baseline and 2, 4, 8 and 12 weeks after injection for Modified Ashworth Score, time walking 10 meters, and the Functional Independence Measure (mFIM) motor score. Results: The higher-dose group exhibited a significant improvement in spasticity, and both groups showed an improvement in time walking 10 meters and mFIM, with no significant differences between them. Conclusions: Our findings suggest that gains in gait velocity and functional independence were not correlated to BTX-A dose. .

O objetivo deste estudo foi avaliar os efeitos da toxina botulínica tipo A (TXB-A) sobre a espasticidade de membro inferior em pacientes pós-AVE em reabilitação. Método: 21 pacientes hemiparéticos foram divididos em dois grupos que receberam doses de TXB-A de 300UI (Grupo 1) e 100UI (Grupo 2) e foram avaliados antes da injeção e 2, 4, 8 e 12 semanas após, quanto à escala de Ashworth modificada, tempo para andar 10 metros e escore motor da Medida de Independência Funcional (MIFm). Resultados: O grupo que utilizou dose mais alta teve melhora significativa da espasticidade. Ambos os grupos tiveram melhora do tempo para andar 10 metros e da MIFm sem diferença significativa entre eles. Conclusões: A melhora da velocidade de marcha e da independência funcional não foram correlacionadas com a dose de TXB-A na amostra analisada. .