Genotoxicity, anti-melanoma and antioxidant activities of Hymenaea courbaril L. seed extract.

Hymenaea courbaril has been used to treat different diseases, although its properties are yet to be scientifically validated. The objective of this study was to determine the cytotoxicity, genotoxicity, antigenotoxicity and antioxidant potentials of hydroethanolic extract from H. courbaril seeds. Therefore, for the cytotoxicity test an anti-melanoma assay was performed in B16F10 strain cells. The genotoxicity and antigenotoxicity was evaluated in bone marrow cells (Permit number: 002/2010) of mice, the antioxidant activity was determined by the DPPH test and the total flavonoid content was also determined. The hydroethanolic extract showed antigenotoxic effect and antioxidant activity. It was verified that total flavonoid content was 442.25±18.03 mg RE/g dry extract. HPLC-PAD chromatogram revealed presence of flavones as majority compound in evaluated extract. The results allowed us to also infer that the hydroethanolic extract from seeds shows cytotoxic activity against B16F10 melanoma cells line and it has dose-and-time-dependency.

Antioxidant and photoprotective potential of Moringa oleifera Lam (Moringaceae) / Potencial antioxidante e fotoprotetor de Moringa oleifera Lam (Moringaceae)

Moringa oleifera Lam. (Moringaceae) is a plant with several biological activities and therapeutic properties. However, the complete knowledge about its pharmacological, biological and ecological effects, and about the active components present in each vegetable part are not still completely elucidated. This study aimed to evaluate the antioxidant and photoprotective activities of different extracts from leaves and flowers of M. oleifera. These activities were assessed through in vitro tests, DPPH radical scavenging method, iron ion chelating effect (FRAP), lipid peroxidation (TBARS), nitric oxide scavenging method and assessment of the activity against the lipid peroxidation through hemolytic method. The photoprotective activity was assessed through spectrophotometric analysis and through in vitro test with Labsphere. It was also determined the extract's phenolic content and total flavonoid through spectrophotometry and HPLC. The obtained results demonstrated that this species have components with antioxidant and photoprotective potential mainly in the extracts obtained from fresh leaves and flowers. Therefore, it was possible to verify that M. oleifera has potential to be used as source of antioxidant components with photoprotective activity mainly due to the presence of phenolic components and among these, the flavonoids.

Moringa oleifera Lam. (Moringaceae) é uma planta com várias atividades biológicas e propriedades terapêuticas. No entanto, o conhecimento completo sobre seus efeitos farmacológicos, biológicos e ecológicos, e sobre os componentes ativos presentes em cada parte vegetal não são ainda completamente elucidados. Este estudo teve como objetivo avaliar as atividades antioxidantes e fotoprotetoras de diferentes extratos de folhas e flores de M. oleifera. Estas atividades foram avaliadas através de testes in vitro, método de eliminação de radicais DPPH, efeito de quelação de íons de ferro (FRAP), peroxidação lipídica (TBARS), método de eliminação de óxido nítrico e avaliação da atividade contra a peroxidação lipídica através do método hemolítico. A atividade fotoprotetora foi avaliada através de análise espectrofotométrica e através de teste in vitro com Labsphere. Também foi determinado o conteúdo fenólico do extrato e o flavonoide total através de espectrofotometria e HPLC. Os resultados obtidos demonstraram que esta espécie possui componentes com potencial antioxidante e fotoprotetor principalmente nos extratos obtidos a partir de folhas frescas e flores. Por conseguinte, foi possível verificar que a M. oleifera tem potencial para ser utilizado como fonte de componentes antioxidantes com atividade fotoprotetora principalmente devido à presença de componentes fenólicos e entre estes, os flavonoides.

Genotoxicity and antioxidant activity of spices and herbs used in brazilian cuisine / Genotoxicidade e atividade antioxidante de especiarias e ervas utilizadas na culinária brasileira

Spices are natural plant products, have been used not only as flavoring and coloring agents, but also as food preservatives and folk medicines throughout the world for thousands of years. Many spices also have been recognized by having both digestive stimulant and carminative actions and also antimicrobial, anti-inflammatory, antimutagenic and anti-carcinogenic potential. Antioxidant and genotoxic potential of species commonly used in Brazil was evaluated. The antioxidant activity was evaluated using different methods, including DPPH radical scavenging activity, ferric reducing power (FRAP), iron ion chelating power, inhibition of lipid peroxidation (TBARS), NO radical scavenging, and oxidative hemolysis inhibition. Furthermore, the antigenotoxic activity was evaluated through mitotic index and chromosome aberration in Allium cepa roots. Quantification of total phenols and flavonoids carried out. The results with the Ocimum basilicum spices in the DPPH test showed activity (82.01%), FRAP (321.12 uM ET and iron chelating activity (94.18) and for the Cinnamomum zeylanicum spice in the TBARS test (18.52%) evaluated by different methods and mechanisms of inactivation of free radicals and according to the evaluation of genotoxicity by the Allium cepa test the spices do not present genotoxic effects.

As especiarias são produtos vegetais naturais, que foram utilizados não só como agentes aromatizantes e colorantes, mas também como conservantes de alimentos e medicamentos populares em todo o mundo há milhares de anos. Muitas especiarias também foram reconhecidas por ter estimulantes digestivos e ações carminativas e também potencial antimicrobiano, anti-inflamatório, antimutagênico e anticarcinogênico. O potencial antioxidante e genotóxico das espécies comumente utilizadas no Brasil foi avaliado. A atividade antioxidante foi avaliada utilizando diferentes métodos, incluindo a atividade de eliminação de radicais DPPH, poder de redução férrica (FRAP), poder quelante de íons de ferro, inibição da peroxidação lipídica (TBARS), eliminação de radicais NO e inibição da hemólise oxidativa. Além disso, a atividade antigenotóxica foi avaliada através do índice mitótico e aberração cromossômica nas raízes do Allium cepa. Quantificação de fenóis totais e flavonoides realizados. Os resultados mostraram que as especiarias (Cinnamomum zeylanicum, Origanum vulgare, Piper nigrum, Zingiber officinale e Ocimum basilicum) apresentaram atividade antioxidante avaliada por diferentes métodos e mecanismos de inativação de radicais livres e de acordo com a avaliação de genotoxicidade pelo teste Allium cepa as especiarias não apresentam efeitos genotóxicos.

Antioxidant effect of Morus nigra on Chagas disease progression.

Considering the widespread popular use of Morus nigra and the amount of scientific information on its antioxidant and anti-inflammatory activity, the effectiveness of this phytotherapeutic compound in the parasitemia progression during the acute phase of Chagas disease and its role in the development of the inflammatory process as well as its effects on the oxidative damage in the chronic phase of infection were evaluated. Thus, 96 male Swiss mice were randomly divided into eight groups, four groups were uninfected controls, and four groups were intraperitoneally infected with 5.0 x 104 blood trypomastigotes forms of T. cruzi QM2 strain. Four batches composed of one uninfected and one infected group were respectively treated with 70% alcohol solution and 25 µL, 50 µL and 75 µL of the phytotherapeutic compound. Levels of antioxidant elements (TBARS, FRAP, GSH and Sulfhydryl groups) were measured in plasma samples. The phytotherapeutic compound's antioxidant activity was measured by polyphenol and total flavonoid quantification, DPPH, NO, and FRAP method. Our results showed that the vehicle influenced some of the results that may have physiological relevance in Chagas disease. However, an important action of M. nigra tincture was observed in the progression of Chagas disease, since our results demonstrated a reduction in parasitemia of treated groups when compared to controls, especially in the group receiving 25 µL. However, in the chronic phase, the 50-µL dosage presented a better activity on some antioxidant defenses and minimized the tissue inflammatory process. Results indicated an important action of M. nigra tincture on the Chagas disease progression.

Serologic and Molecular Evidence of Vaccinia Virus Circulation among Small Mammals from Different Biomes, Brazil.

Vaccinia virus (VACV) is a zoonotic agent that causes a disease called bovine vaccinia, which is detected mainly in milking cattle and humans in close contact with these animals. Even though many aspects of VACV infection have been described, much is still unknown about its circulation in the environment and its natural hosts/reservoirs. To investigate the presence of Orthopoxvirus antibodies or VACV DNA, we captured small rodents and marsupials in 3 areas of Minas Gerais state, Brazil, and tested their samples in a laboratory. A total of 336 animals were tested; positivity ranged from 18.1% to 25.5% in the 3 studied regions located in different biomes, including the Atlantic Forest and the Cerrado. Analysis of nucleotide sequences indicated co-circulation of VACV groups I and II. Our findings reinforce the possible role played by rodents and marsupials in VACV maintenance and its transmission chain.

Detection of Vaccinia Virus in Urban Domestic Cats, Brazil.

We investigated possible vaccinia virus (VACV) in urban house cats in Brazil. Serum samples from 6 cats were positive for VACV by PCR, indicating likely VACV circulation among house cats in urban areas of Brazil. This finding highlights the importance of epidemiologic surveillance to avoid outbreaks among urban human populations.

Detection of Vaccinia Virus in Dairy Cattle Serum Samples from 2009, Uruguay.

We detected orthopoxvirus in 28 of 125 serum samples collected during 2009 from cattle in Uruguay. Two samples were PCR-positive for vaccinia virus and had sequences similar to those for vaccinia virus associated with outbreaks in Brazil. Autochthonous circulation of vaccinia virus in Uruguay and other South American countries cannot be ruled out.

Seroprevalence of Orthopoxvirus in rural Brazil: insights into anti-OPV immunity status and its implications for emergent zoonotic OPV.

BACKGROUND: Bovine vaccinia (BV) is a zoonosis caused by Vaccinia virus, a virus from Orthopoxvirus genus (OPV) that affects mainly cattle herds and humans in rural areas in Brazil. Because most studies have focused on outbreaks situations, data on BV epidemiology is limited. A cross sectional study in Brazilian rural areas during 2012-2013 was conducted to determine the neutralizing antibodies seroprevalence and risk factors for BV. METHODS: A structured questionnaire was applied to elicit demographics data and farming practices considered risk factors for BV exposure. Neutralizing anti-OPV antibodies were investigated using plaque reduction neutralization test. The neutralizing antibodies prevalence rates were calculated and the risk factor analysis was performed using multivariate logistic regression. RESULTS: Two hundred and forty participants were enrolled in this study with a prevalence of neutralizing antibodies of 30.8 % (95 % confidence interval [CI], 25.3-36.9). In multivariate analysis, age > 35 years (Odds Ratio [OR] = 18.2; CI 95 % = 7.7 - 43.2) and previous outbreak in property (OR = 3.9; C I95 % = 1.2 - 12.6) were independently associated with anti-OPV neutralizing antibodies. CONCLUSIONS: In this study, anti-OPV protective immunity (neutralizing antibody titers) was assessed in an endemic BV Brazilian rural area. Our findings indicate that epidemiological surveillance is required and should be applied by public health authorities to create interventions and/or prevention strategies to avoid viral spread causing future outbreaks among individuals who are under risk of infection.

The detection of Vaccinia virus confirms the high circulation of Orthopoxvirus in buffaloes living in geographical isolation, Marajó Island, Brazilian Amazon.

In Brazil, serologic evidence of Orthopoxvirus (OPV) circulation showed positivity around 20% in cattle, humans, monkeys and rodents. Although OPV seropositivity has been described in buffalo herds in southeastern Brazil, no Vaccinia virus (VACV) (member of genus OPV) outbreaks in buffalo herds have been described in this country. This study aimed to investigate the detection of anti-OPV antibodies and to study the OPV genome in Brazilian buffalo herds. Our results demonstrated a high OPV seropositivity in buffalo herds on Marajó Island and molecular data confirmed the circulation of VACV. The geographical isolation conditionmight be a sine qua non condition to explain our results.

Infection of the central nervous system with dengue virus 3 genotype I causing neurological manifestations in Brazil.

A case of dengue virus 3 (DENV-3) genotype I infection with neurological manifestations occurred in Belo Horizonte, Minas Gerais in October 2012. The serotype was detected by PCR, and the genotype was assessed by sequencing and phylogenetic analysis of the C-prM region. The virus causing neurological manifestations clustered with other sequences of DENV-3 genotype I. Because neurological manifestations of DENV are possibly misdiagnosed in Brazil, this study serves as an alert of the importance of DENV diagnoses in CNS infections.

Infection of the central nervous system with dengue virus 3 genotype I causing neurological manifestations in Brazil

Abstract: A case of dengue virus 3 (DENV-3) genotype I infection with neurological manifestations occurred in Belo Horizonte, Minas Gerais in October 2012. The serotype was detected by PCR, and the genotype was assessed by sequencing and phylogenetic analysis of the C-prM region. The virus causing neurological manifestations clustered with other sequences of DENV-3 genotype I. Because neurological manifestations of DENV are possibly misdiagnosed in Brazil, this study serves as an alert of the importance of DENV diagnoses in CNS infections.

Evaluating anti-Orthopoxvirus antibodies in individuals from Brazilian rural areas prior to the bovine vaccinia era.

Vaccinia virus naturally circulates in Brazil and is the causative agent of a zoonotic disease known as bovine vaccinia (BV). We retrospectively evaluated two populations from the Amazon and Southeast Regions. BV outbreaks had not been reported in these regions before sample collection. Neutralising antibodies were found in 13 individuals (n = 132) with titres ranging from 100 ≥ 6,400 neutralising units/mL. Univariate analysis identified age and vaccination as statistically significant risk factors in individuals from the Southeast Region. The absence of detectable antibodies in vaccinated individuals raises questions about the protection of smallpox vaccine years after vaccination and reinforces the need for surveillance of Orthopoxvirus in Brazilian populations without evidence of previous outbreaks.

RAP1 GTPase overexpression is associated with cervical intraepithelial neoplasia.

RAP1 (RAS proximate 1), a small GTP-binding protein of the RAS superfamily, is a putative oncogene that is highly expressed in several malignant cell lines and types of cancers, including some types of squamous cell carcinoma. However, the participation of RAP1 in cervical carcinogenesis is unknown. We conducted a cross-sectional study of paraffin-embedded cervical biopsies to determine the association of RAP1 with cervical intraepithelial neoplasia (CIN). Standard and quantitative immunohistochemistry assessment of RAP1 expression in fixed tissue was performed on 183 paraffin-embedded cervical biopsies that were classified as normal or non-dysplastic mucosa (NDM) (n = 33); CIN grade 1 (n = 84) and CIN grade 2/3 (n = 66). A gradual increase in RAP1 expression in NDM < CIN 1 < CIN 2/3 (p<0.001) specimens was observed and was in agreement with the histopathologic diagnosis. A progressive increase in the RAP1 expression levels increased the risk of CIN 1 [odds ratio (OR) = 3.50; 95% confidence interval (CI) 1.30-10.64] 3.5 fold and the risk of CIN 2/3 (OR = 19.86, 95% CI 6.40-70.79) nearly 20 fold when compared to NDM. In addition, stereotype ordinal regression analysis showed that this progressive increase in RAP1 expression more strongly impacted CIN 2/3 than CIN 1. Our findings suggest that RAP1 may be a useful biomarker for the diagnosis of CIN.

Outbreak of severe zoonotic vaccinia virus infection, Southeastern Brazil.

In 2010, a vaccinia virus isolate caused an atypically severe outbreak that affected humans and cattle in Brazil. Of 26 rural workers affected, 12 were hospitalized. Our data raise questions about the risk factors related to the increasing number and severity of vaccinia virus infections.

From lesions to viral clones: biological and molecular diversity amongst autochthonous Brazilian vaccinia virus.

Vaccinia virus (VACV) has had an important role for humanity because of its use during the smallpox eradication campaign. VACV is the etiologic agent of the bovine vaccinia (BV), an emerging zoonosis that has been associated with economic, social, veterinary and public health problems, mainly in Brazil and India. Despite the current and historical VACV importance, there is little information about its circulation, prevalence, origins and maintenance in the environment, natural reservoirs and diversity. Brazilian VACV (VACV-BR) are grouped into at least two groups based on genetic and biological diversity: group 1 (G1) and group 2 (G2). In this study, we went to the field and investigated VACV clonal diversity directly from exanthemous lesions, during BV outbreaks. Our results demonstrate that the G1 VACV-BR were more frequently isolated. Furthermore, we were able to co-detect the two variants (G1 and G2) in the same sample. Molecular and biological analysis corroborated previous reports and confirmed the co-circulation of two VACV-BR lineages. The detected G2 clones presented exclusive genetic and biological markers, distinct to reference isolates, including VACV-Western Reserve. Two clones presented a mosaic profile, with both G1 and G2 features based on the molecular analysis of A56R, A26L and C23L genes. Indeed, some SNPs and INDELs in A56R nucleotide sequences were observed among clones of the same virus population, maybe as a result of an increased mutation rate in a mixed population. These results provide information about the diversity profile in VACV populations, highlighting its importance to VACV evolution and maintenance in the environment.

Caraparu virus induces damage and alterations in antioxidant defenses in the liver of BALB/c mice after subcutaneous infection.

Oxidative stress is a disturbance in the oxidant-antioxidant balance leading to potential cellular damage. Most cells can tolerate a mild degree of oxidative stress because they have a system that counteracts oxidation that includes antioxidant molecules such as glutathione (GSH) and superoxide dismutase (SOD). Disruption of the host antioxidant status has been recognized as an important contributor to the pathogenesis of many viruses. Caraparu virus (CARV) is a member of group C of the Bunyaviridae family of viruses. In South American countries, group C bunyaviruses are among the common agents of human febrile illness and have caused multiple notable outbreaks of human disease in recent decades; nevertheless, little is known about the pathogenic characteristics of these viruses. The purpose of this study was to examine the hepatic pathogenesis of CARV in mice and the involvement of oxidative stress and antioxidant defenses on this pathology. Following subcutaneous infection of BALB/c mice, CARV was detected in the liver, and histopathology revealed acute hepatitis. Increased serum levels of aspartate and alanine aminotransferases (AST/ALT) and greater hepatic expression of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) were found in infected animals. CARV infection did not alter the biomarkers of oxidative stress but caused an increase in GSH content and altered the expression and activity of SOD. This is the first report of an alteration of oxidative homeostasis upon CARV infection, which may, in part, explain the hepatic pathogenesis of this virus, as well as the pathogenesis of other Bunyaviridae members.

Differential upregulation of human 2'5'OAS genes on systemic sclerosis: Detection of increased basal levels of OASL and OAS2 genes through a qPCR based assay.

2'5'OAS are template-independent RNA polymerases with antiviral activity and important to homeostasis maintenance. Here we have developed quantitative PCR (qPCR) reactions for the detection of each individual 2'5'OAS human gene and used them to evaluate these gene levels in systemic sclerosis patients cells. The method was efficient for quantification of 2'5'OAS genes on human cells after interferon (IFN) treatment, and revealed that primary cells from patients with systemic sclerosis have increased basal levels of OASL and OAS2 genes. When treated, patients cells are able to induce all four 2'5'OAS genes. Our hypothesis is that abnormally circulating type I IFNs on the disease could be establishing a desensitized state on patients cells, making them refractory to subsequent IFN doses, and that OASL and OAS2 genes upregulation may be due to an IFN-independent stimulus. Further characterizing the biological activities of these genes, their induction pathways and their regulatory functions can lead to better understanding of systemic sclerosis molecular mechanisms and of their biological activities.