Pedunculagin and tellimagrandin-I stimulate inflammation and angiogenesis and upregulate vascular endothelial growth factor and tumor necrosis factor-alpha in vivo.

Pedunculagin (PD) and tellimagrandin-I (TL), isolated from Myrciaria cauliflora seeds and Eucaliptus microcorys leaves, respectively, have attracted great attention owing to their relevant biological activities, such as antitumor, antioxidant, and hepatoprotective activities. This study investigated the angiogenic potential of PD and TL using a chick embryo chorioallantoic membrane (CAM) assay. Using the CAM assay, our results showed that both PD and TL promoted a significant increase in the number and caliber of blood vessels, the thickness of the CAM, and the presence of fibroblasts and inflammatory cells. Moreover, an increase of tumor necrosis factor-α and vascular endothelial growth factor was observed in the CAM treated with PD and TL, indicating the induction of angiogenic factors. Thus, the remarkable profile of PD and TL in inducing angiogenesis opens up new perspectives for their potential utilization in different therapeutic approaches involving neovascularization.

The presence of antibiotics and multidrug-resistant Staphylococcus aureus reservoir in a low-order stream spring in central Brazil.

The disposal of industrial effluents strongly influences low-order streams, which makes them fragile ecosystems that can be impacted by contamination. In central Brazil, the Extrema River spring targets the dumping of pharmaceutical products from the surrounding industries. So, this work aimed to investigate the presence of antibiotics in Extrema River spring samples and the isolation of Staphylococcus aureus, a potential multidrug-resistant bacteria, verifying the antimicrobial resistance profile of these isolates. Three campaigns were carried out in different locals (P1-P3) between October and December 2021, in the dry and rainy seasons. The high-performance liquid chromatography-tandem mass spectrometry (LCMS) approach indicated the presence of sulfamethoxazole (≥ 1 ng/L), metronidazole (< 0.5 ng/L), and chloramphenicol (< 5 ng/L) in the water samples in November (rainy season). S. aureus was isolated in P1 (n = 128), P2 (n = 168), and P3 (n = 36), with greater resistance to trimethoprim-sulfamethoxazole (90%), clindamycin (70%), and gentamicin (60%). The presence of antibiotics in the Extrema River spring may cause S. aureus antibiotic resistance development. The presence of antibiotics and the high percentage of isolated multidrug-resistant S. aureus in the Extrema River spring cause concern and indicate the clandestine dumping of effluents from nearby pharmaceutical industries. Since preserving the springs of low-order streams is important for the environment and public health, we encourage monitoring the wastewater from Extrema River's nearby pharmaceutical industries and preserving the spring of this river.

Genotoxicity and maternal-fetal safety of the dried extract of leaves of Azadirachta indica A. Juss (Meliaceae) in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Azadirachta indica A. Juss (Meliaceae), popularly known as "neem", is used for the treatment of rheumatism, cancer, ulcers, diabetes, respiratory problems, among others. This species is present on six continents and contains more than 400 bioactive compounds. Practically all parts of the plant are used in the treatment of diseases. Although it is widely used, no study has evaluated the safety of this species throughout the gestational period in Wistar rats. AIM OF THE STUDY: To evaluate the genotoxicity and the effect of treatment with dried extract of leaves of Azadirachta indica on maternal toxicity and fetal development. MATERIALS AND METHODS: The dried extract of leaves of A. indica was obtained by spray drying after percolation of the plant material in 30% ethanol (w/w). The total flavonoids and rutin contents of the extract were determined by spectrophotometric method and HPLC-DAD, respectively. Pregnant Wistar rats (n = 40) were divided into four groups (n = 10/group): one control and three groups treated with dried extract of leaves of A. indica at doses of 300, 600 or 1200 mg/kg. Treatments were carried out from gestational day (GD) 0-20. During gestation, clinical signs of toxicity, weight gain, feed and water consumption of the dams were evaluated. On GD 21, rats were euthanized and cardiac blood was collected. Liver, kidneys, lung, heart, uterus, ovaries and bone marrow were collected. Reproductive performance parameters, histopathological analysis, biochemistry and genotoxicity were evaluated. Fetuses were evaluated for external morphology, skeletal and visceral changes. RESULTS: The total flavonoid content of the extract ranged from 2.64 to 3.01%, and the rutin content was 1.07%. There was no change in body mass gain, food and water consumption between the evaluated groups. There was also no difference between the groups in terms of biochemical parameters, reproductive performance, histopathological analysis of the mother's organs and genotoxicity. Supernumerary ossification sites of the sternum were observed, and other skeletal and visceral alterations were not significant. CONCLUSIONS: The treatment did not induce maternal toxicity, it was neither embryotoxic nor fetotoxic. The extract was not potentially genotoxic, and at a dose of 1200 mg/kg, it caused changes in the ossification of the sternum.

Effects of liquid extract from Plinia cauliflora fruits residues on Chinese hamsters biochemical parameters / Efeitos do extrato líquido de resíduos de frutos de Plinia cauliflora sobre os parâmetros bioquímicos de hamsters chineses

Plinia cauliflora (Mart.) Kausel, popularly known as jabuticaba, is rich in polyphenols. Phenolic compounds exhibit several biological properties, which reflect on biomarkers such as biochemical parameters. In the present study, we evaluated the plasmatic levels of glucose, total cholesterol, HDL-cholesterol, triglycerides, and uric acid of Chinese hamsters fed for 45 days with a regular diet or cholesterol-enriched diet supplemented with a liquid extract obtained from P. cauliflora fruits residues standardized in ellagic acid and total phenolic compounds. The results showed that the concentrated extract obtained from jabuticaba residues increased the glycemia of animals fed with a regular diet and reduced the plasmatic uric acid levels of animals fed with a cholesterol-enriched diet. Since hyperuricemia is considered to be a significant risk factor of metabolic disorders and the principal pathological basis of gout, the liquid extract from P. cauliflora fruits residues would be a promising candidate as a novel hypouricaemic agent for further investigation.

Plinia cauliflora (Mart.) Kausel, popularmente conhecida como jabuticaba, é rica em polifenois. Os compostos fenólicos apresentam diversas propriedades biológicas, que refletem em biomarcadores, como os parâmetros bioquímicos. No presente estudo, avaliamos os níveis plasmáticos de glicose, colesterol total, HDL-colesterol, triglicerídeos e ácido úrico em hamsters chineses alimentados por 45 dias com dieta regular ou dieta enriquecida com colesterol suplementada com extrato líquido obtido de resíduos de frutos de P. cauliflora padronizado em ácido elágico e compostos fenólicos totais. Os resultados mostraram que o extrato concentrado obtido dos resíduos de jabuticaba aumentou a glicemia dos animais alimentados com dieta regular e reduziu os níveis plasmáticos de ácido úrico dos animais alimentados com dieta rica em colesterol. Uma vez que a hiperuricemia é considerada um fator de risco significativo de distúrbios metabólicos e a principal base patológica da gota, o extrato líquido dos resíduos de frutas de P. cauliflora seria um candidato promissor como um novo agente hipouricêmico para investigação posterior.

Effects of liquid extract from Plinia cauliflora fruits residues on Chinese hamsters biochemical parameters

Abstract Plinia cauliflora (Mart.) Kausel, popularly known as jabuticaba, is rich in polyphenols. Phenolic compounds exhibit several biological properties, which reflect on biomarkers such as biochemical parameters. In the present study, we evaluated the plasmatic levels of glucose, total cholesterol, HDL-cholesterol, triglycerides, and uric acid of Chinese hamsters fed for 45 days with a regular diet or cholesterol-enriched diet supplemented with a liquid extract obtained from P. cauliflora fruits residues standardized in ellagic acid and total phenolic compounds. The results showed that the concentrated extract obtained from jabuticaba residues increased the glycemia of animals fed with a regular diet and reduced the plasmatic uric acid levels of animals fed with a cholesterol-enriched diet. Since hyperuricemia is considered to be a significant risk factor of metabolic disorders and the principal pathological basis of gout, the liquid extract from P. cauliflora fruits residues would be a promising candidate as a novel hypouricaemic agent for further investigation.

Resumo Plinia cauliflora (Mart.) Kausel, popularmente conhecida como jabuticaba, é rica em polifenois. Os compostos fenólicos apresentam diversas propriedades biológicas, que refletem em biomarcadores, como os parâmetros bioquímicos. No presente estudo, avaliamos os níveis plasmáticos de glicose, colesterol total, HDL-colesterol, triglicerídeos e ácido úrico em hamsters chineses alimentados por 45 dias com dieta regular ou dieta enriquecida com colesterol suplementada com extrato líquido obtido de resíduos de frutos de P. cauliflora padronizado em ácido elágico e compostos fenólicos totais. Os resultados mostraram que o extrato concentrado obtido dos resíduos de jabuticaba aumentou a glicemia dos animais alimentados com dieta regular e reduziu os níveis plasmáticos de ácido úrico dos animais alimentados com dieta rica em colesterol. Uma vez que a hiperuricemia é considerada um fator de risco significativo de distúrbios metabólicos e a principal base patológica da gota, o extrato líquido dos resíduos de frutas de P. cauliflora seria um candidato promissor como um novo agente hipouricêmico para investigação posterior.

What can the Allium cepa test say about pesticide safety? A review.

The increasing use of pesticides has caused global concerns about the toxic effects and adverse consequences of pesticides on humans and the environment. Among the ways to understand the impact of pesticides, the Allium cepa bioassay stands out. This test is suitable to evaluate different toxic, cytotoxic, genotoxic, and mutagenic outcomes. In this context, the present review aimed to summarize the history of using the A. cepa bioassay to investigate pesticide damages. Data on the experimental conditions were also discussed. The reviewed studies showed the toxicity profile of 113 active ingredients primarily tested in the laboratory, using water for exposure. The most used biomarkers were the mitotic index, chromosomal aberrations, and nuclear abnormalities. All active ingredients caused some toxicity levels in A. cepa, showing the efficiency and sensibility of this bioindicator and the adverse effect of pesticides on humans and the environment. Furthermore, it was evident that pesticides have great potential to damage the mitotic spindle and DNA because almost all active ingredients tested induced chromosomal aberrations and nuclear abnormalities. The current review showed that the A. cepa bioassay is an effective and appropriate model to evaluate pesticide toxicity, and it might indicate research gaps and recommendations for further studies.

Toxic Potential of Cerrado Plants on Different Organisms.

Cerrado has many compounds that have been used as biopesticides, herbicides, medicines, and others due to their highly toxic potential. Thus, this review aims to present information about the toxicity of Cerrado plants. For this purpose, a review was performed using PubMed, Science Direct, and Web Of Science databases. After applying exclusion criteria, 187 articles published in the last 20 years were selected and analyzed. Detailed information about the extract preparation, part of the plant used, dose/concentration tested, model system, and employed assay was provided for different toxic activities described in the literature, namely cytotoxic, genotoxic, mutagenic, antibacterial, antifungal, antiviral, insecticidal, antiparasitic, and molluscicidal activities. In addition, the steps to execute research on plant toxicity and the more common methods employed were discussed. This review synthesized and organized the available research on the toxic effects of Cerrado plants, which could contribute to the future design of new environmentally safe products.

Phytochemical Composition and Protective Effect of Vernonanthura polyanthes Leaf against In Vivo Doxorubicin-Mediated Toxicity.

Vernonanthura polyanthes (Spreng.) A.J. Vega & Dematt. (syn.: Vernonia polyanthes Less) is popularly known as "assa-peixe" and its leaves are used in folk medicine mainly to treat respiratory diseases. In this study, we evaluated the cytogenotoxic and anticytogenotoxic potential of the V. polyanthes leaf aqueous extract (VpLAE) and its n-butanol fraction (n-BF) in the presence or absence of doxorubicin (DXR) (pre-, co-, and post-treatments) on a murine model for 24 h or 120 h. The micronucleus test (MN) and the comet assay were used to assess the cytogenotoxic and anticytogenotoxic potential of VpLAE and n-BF (250, 500, and 1000 mg/kg) administered via gavage to Swiss Webster mice. The chemical profiles of VpLAE and n-BF were assessed by liquid chromatography coupled to mass spectrometry, and their metabolites were putatively identified. Lastly, the possible biological activities related to the (anti) cytogenotoxicity of the compounds were predicted using the PASS online webserver. The in vivo results showed that different doses of VpLAE and n-BF did not present cytotoxic activity; however, the MN test revealed a slight mutagenic activity for the 24 h treatments. Moderate genotoxic effects were demonstrated for all treatments in the comet assay. Regarding anticytotoxicity and antimutagenicity, VpLAE and n-BF presented a high cytoprotective potential against DXR toxic effects. In the co-treatment, VpLAE reduced the DXR genotoxicity by ~27%, and n-BF did not demonstrate antigenotoxic potential. In contrast, an antigenotoxic effect was observed for both VpLAE and n-BF in the pre- and post-treatments, reducing DXR genotoxicity by ~41% and ~47%, respectively. Chemical analysis of VpLAE and n-BF showed the presence of eight phenolic compounds, including seven chlorogenic acids and a flavonoid. The PASS online tool predicted antimutagenic, anticancer, antineoplastic, chemoprotective, antioxidant, and radical scavenging activities for all constituents identified in VpLAE and n-BF. V. polyanthes leaves presented a protective effect against DXR cytogenotoxicity. In general, VpLAE and n-BF showed a greater antigenotoxic potential in the pre- and post-treatments. The metabolites putatively identified in VpLAE and n-BF exhibited antioxidant and chemoprotective potential according to computational prediction analysis. Altogether, our results highlight the potential application of V. polyanthes to protect against toxic manifestations induced by DXR.

Micronucleus test and nuclear abnormality assay in zebrafish (Danio rerio): Past, present, and future trends.

Nuclear abnormality (NA) assay in fish has been widely applied for toxicity risk assessment under field and laboratory conditions. The zebrafish (Danio rerio) has become a suitable model system for assessing the NA induced by pollutants. Thus, the current study aimed to summarize and discuss the literature concerning micronucleus (MN) and other NA in zebrafish and its applications in toxicity screening and environmental risk assessment. The data concerning the publication year, pollutant type, experimental design, and type of NA induced by pollutants were summarized. Also, molecular mechanisms that cause NA in zebrafish were discussed. Revised data showed that the MN test in zebrafish has been applied since 1996. The MN was the most frequently NA, but 15 other nuclear alterations were reported in zebrafish, such as notched nuclei, blebbed nuclei, binucleated cell, buds, lobed nuclei, bridges, and kidney-shaped. Several pollutants can induce NA in zebrafish, mainly effluents (mixture of pollutants), agrochemicals, and microplastics. The pollutant-induced NA in zebrafish depends on experimental design (i.e., exposure time, concentration, and exposure condition), developmental stages, cell/tissue type, and the type of pollutant. Besides, research gaps and recommendations for future studies are indicated. Overall, the current study showed that zebrafish is a suitable model to assess pollutant-induced mutagenicity.

Effects of liquid extract from Plinia cauliflora fruits residues on Chinese hamsters biochemical parameters.

Plinia cauliflora (Mart.) Kausel, popularly known as jabuticaba, is rich in polyphenols. Phenolic compounds exhibit several biological properties, which reflect on biomarkers such as biochemical parameters. In the present study, we evaluated the plasmatic levels of glucose, total cholesterol, HDL-cholesterol, triglycerides, and uric acid of Chinese hamsters fed for 45 days with a regular diet or cholesterol-enriched diet supplemented with a liquid extract obtained from P. cauliflora fruits residues standardized in ellagic acid and total phenolic compounds. The results showed that the concentrated extract obtained from jabuticaba residues increased the glycemia of animals fed with a regular diet and reduced the plasmatic uric acid levels of animals fed with a cholesterol-enriched diet. Since hyperuricemia is considered to be a significant risk factor of metabolic disorders and the principal pathological basis of gout, the liquid extract from P. cauliflora fruits residues would be a promising candidate as a novel hypouricaemic agent for further investigation.

Acute exposure to environmentally relevant concentrations of benzophenone-3 induced genotoxicity in Poecilia reticulata.

The organic UV filter benzophenone-3 (BP-3), widely used in the commercial formulations of sunscreens and personal care products, is considered an emerging pollutant and has been associated with several human and environmental health concerns. However, knowledge about their mode of action and ecotoxicity on aquatic biota is scarce. In this scenario, the objective of this work was to evaluate the genotoxic, mutagenic, and erythrotoxicity effects of BP-3 in the guppy Poecilia reticulata after acute exposure. Adult females of P. reticulata were exposed to three non-lethal and environmentally relevant concentrations of BP-3 (10, 100, and 1000 ng L-1) during 96 h of exposure, and the somatic parameter [Fulton condition factor (K)], genotoxicity (comet assay), mutagenicity [micronucleus (MN) and erythrocyte nuclear abnormalities (ENA) tests] and erythrotoxicity parameters (such as total cell area and nucleus-cytoplasmic ratio) were analyzed. Results showed that the general physiological condition (K value) of fish was not affected by acute exposure to BP-3. However, BP-3 induced DNA damage at 100 and 1000 ng L-1 and increased the frequency of total ENA at 1000 ng L-1, specially lobed nucleus, when compared to control group, indicating its genotoxic and mutagenic effects. Furthermore, the BP-3 did not induce significant changes in the total cell area and nucleus-cytoplasmic ratio. In summary, results showed that the BP-3 at environmentally relevant concentration was genotoxic to freshwater fish P. reticulata, confirming its environmental risk.

The chick embryo chorioallantoic membrane assay as a model for the study of angiogenesis / Ensaio da membrana corioalantóide de embrião de galinha como modelo para o estudo da angiogênese

Angiogenesis is a fundamental physiological process with strong implications in tissue homeostasis. Animal models helping to identify how angiogenesis is regulated are fundamental to answer many biological questions. Chick embryo chorioallantoic membrane (CAM) assay is one of the most employed methods to study angiogenesis. In this study we applied a scientometric approach to evaluate the employment of CAM assay in published articles. Temporal trends indicated that CAM assay was the preferred method to investigate angiogenesis over time. The publications had a significant number of citations and the impact factor of journals publishing articles is relevant for the scientific community. A total of 52 different research areas have articles published using this particular technique. Oncology is the research field in which CAM assay was mostly used. Accordingly, tumor-derived cell lines were the most frequent sample tested on CAM. We also identified that 73,6% of articles published used only CAM assay to answer questions concerning angiogenesis. We concluded that although the CAM assay is a classical approach, that does not need so much infrastructure and financial support to be performed, it is a well-accepted technique by the scientific community. In addition, this methodology has gain attention in scientific community because no pain is experienced by the chick and they are minor ethical concerns to employ this method. Moreover, this data can help researchers who are unfamiliar with the CAM assay to identify if this particular method is suitable for their research.

A angiogênese é um processo fisiológico fundamental com fortes implicações na homeostase tecidual. Modelos animais que ajudam a entender como a angiogênese é regulada, são fundamentais para responder a muitas questões biológicas. O ensaio de membrana corioalantóide de embrião de galinha (CAM) é um dos métodos mais empregados para estudar a angiogênese. Neste estudo foi aplicada uma abordagem cientométrica para avaliar o emprego do ensaio CAM em artigos científicos já publicados. Tendências temporais indicaram que o ensaio CAM foi o método mais usado para investigar a angiogênese ao longo do tempo. Os artigos científicos que usaram a metodologia CAM foram publicados em periódicos com significativos números de citações e fator de impacto. No total 52 diferentes áreas de conhecimento usaram a técnica CAM, sendo a oncologia o campo o qual produziu maior número de artigos usando essa metodologia. Consequentemente o material biológico mais testado foi as linhagens celulares tumorais. Também foi identificado que 73,6% dos artigos publicados utilizaram apenas o teste CAM para responder questões relacionadas à angiogênese. Pode se concluir que embora o ensaio CAM seja uma abordagem clássica, que não necessita de muita infraestrutura e apoio financeiro para ser realizado, é uma técnica bem aceita pela comunidade científica. Além disso, esta metodologia tem ganhado atenção na comunidade científica porque os animais testados não sofrem dor e por essa razão esse modelo experimental exige mínimas preocupações éticas. Além disso, esses dados podem ajudar os pesquisadores que não estão familiarizados com o ensaio CAM a identificar se esse método específico é adequado para sua pesquisa.

Antiangiogenic potential of Jatropha curcas latex in the chick chorioallantoic membrane model / Potencial antiangiogênico do látex de Jatropha curcas em modelo de membrana corioalantoica de embrião de galinha

AIMS: To perform a physicochemical and phytochemical characterization of Jatropha curcas latex and to investigate its antiangiogenic potential. METHODS: We performed an initial physicochemical characterization of J. curcas latex using thermal gravimetric analyses and Fourier Transform Infrared spectroscopy. After that, phenols, tannins and flavonoids were quantified. Finally, the potential of J. curcas latex to inhibit angiogenesis was evaluated using the chick chorioallantoic membrane model. Five groups of 20 fertilized chicken eggs each had the chorioallantoic membrane exposed to the following solutions: (1) water, negative control; (2) dexamethasone, angiogenesis inhibitor; (3) Regederm®, positive control; (4) 25% J. curcas latex diluted in water; (5) 50% J. curcas latex diluted in water; and (6) J. curcas crude latex. Analysis of the newly-formed vascular net was made through captured images and quantification of the number of pixels. Histological analyses were performed to evaluate the inflammation, neovascularization, and hyperemia parameters. The results were statically analyzed with a significance level set at p<0.05. RESULTS: Physicochemical characterization showed that J. curcas latex presented a low amount of cis-1.4-polyisoprene, which reduced its elasticity and thermal stability. Phytochemical analyses of J. curcas latex identified a substantial amount of phenols, tannins, and flavonoids (51.9%, 11.8%, and 0.07% respectively). Using a chick chorioallantoic membrane assay, we demonstrated the antiangiogenic potential of J. curcas latex. The latex induced a decrease in the vascularization of the membranes when compared with neutral and positive controls (water and Regederm®). However, when compared with the negative control (dexamethasone), higher J. curcas latex concentrations showed no significant differences. CONCLUSIONS: J. curcas latex showed low thermal stability, and consisted of phenols, tannins, and flavonoids, but little or no rubber. Moreover, this latex demonstrated a significant antiangiogenic activity on a chick chorioallantoic membrane model. The combination of antimutagenic, cytotoxic, antioxidant and antiangiogenic properties makes J. curcas latex a potential target for the development of new drugs.

OBJETIVOS: Realizar uma caracterização físico-química e fitoquímica do látex de Jatropha curcas e investigar o seu potencial antiangiogênico. MÉTODOS: foi realizada uma caracterização físico-química inicial do látex de J. curcas utilizando as análises termogravimétricas e a espectroscopia com a Transformada de Fourier. Depois disso, fenóis, taninos e flavonoides foram quantificados. Finalmente, o potencial do látex de J. curcas em inibir a angiogênese foi avaliado através do uso de modelo de membrana corioalantoica de embrião de galinha. Cinco grupos, cada um com 20 ovos de galinha fertilizados, tiveram a membrana corioalantoica exposta às seguintes soluções: (1) água, controle negativo; (2) dexametasona, inibidor da angiogênese; (3) Regederm®, controle positivo; (4) 25% de látex de J. curcas diluído em água; (5) 50% de látex de J. curcas diluído em água; e (6) látex bruto de J. curcas. A análise da rede vascular recém-formada foi feita por meio de imagens capturadas e quantificação do número de pixels. Análises histológicas foram realizadas para avaliar os parâmetros de inflamação, neovascularização e hiperemia. Os resultados foram analisados estaticamente com nível de significância estabelecido em p<0,05. RESULTADOS: A caracterização físico-química mostrou que o látex de J. curcas apresenta uma baixa quantidade de cis-1,4-poliisopreno, o que reduz sua elasticidade e estabilidade térmica. Análises fitoquímicas do látex de J. curcas identificaram uma quantidade significativa de fenóis, taninos e flavonoides (51,9%, 11,8% e 0,07% respectivamente). Usando o modelo de membrana corioalantoica de ovo de galinha embrionado, demonstrou-se o potencial antiangiogênico do látex de J. curcas. O látex induziu a diminuição da vascularização das membranas, em comparação aos grupos controle neutro e positivo (água e Regederm®). CONCLUSÕES: O látex de J. curcas apresentou baixa estabilidade térmica, ausência ou pouca quantidade de borracha e presença de fenóis, taninos e flavonoides em sua composição. Além disso, apresentou alta atividade antiangiogênica no modelo de membrana corioalantoica de embrião de galinha. A combinação de propriedades antimutagênicas, citotóxicas, anti-inflamatórias, antioxidantes e antiangiogênicas faz com que o látex de J. curcas seja um alvo potencial para o desenvolvimento de novos medicamentos.

Toxicity of Jatropha curcas L. latex in Allium cepa test / Toxicidade do látex de Jatropha curcas L. no modelo de Allium cepa

The latex obtained from Jatropha curcas (physic nut) is used in traditional medicine to treat a variety of disturbs, including burns, hemorrhoids, ringworm and ulcers. Phytochemical analyses have shown that J. curcas latex contains natural compounds with therapeutic potential. In this study, the toxicity, cytotoxicity and genotoxicity effects of J. curcas latex on the root cells of Allium cepa were examined. Onion seeds and bulbs were exposed to seven different concentrations of latex and then the roots were submitted to macro and microscopic analyses. Water and sodium azide were used as negative and positive controls, respectively. The analysis of root growth showed that J. curcas crude latex or 50% diluted is highly toxic. Cytogenetic results showed that the mitotic index of the onion roots submitted to latex treatment decreased significantly compared to the negative control, which suggests that the latex is cytotoxic. High incidence of chromosome aberrations in the cells treated with J. curcas latex was observed too, indicating that the latex also presents genotoxic effect. The analyses presented in this report suggest the toxic, cytotoxic and genotoxic effects of J. curcas latex. Then, the indiscriminate use of J. curcas latex in folk medicine could bring risk to human health.

O látex obtido de Jatropha curcas (pinhão manso) é usado na medicina tradicional para tratamento de diversos distúrbios, como queimaduras, hemorroida, micose e úlcera. Análises fitoquímicas apontaram que o látex de J. curcas contém compostos naturais com potencial terapêutico. Este estudo avaliou a toxicidade, citotoxicidade e genotoxicidade do látex de J. curcas em células da raiz de Allium cepa. Sementes e bulbos de cebola foram expostos á sete diferentes concentrações de látex e, então, as raízes foram submetidas a análises macro e microscópica. Água e azida sódica foram utilizadas como controle negativo e positivo, respectivamente. A análise do comprimento das raízes mostrou que o látex de J. curcas puro e diluído a 50% é altamente tóxico. O índice mitótico das raízes de cebola submetidas ao tratamento com o látex diminuiu significativamente comparado com o controle negativo, o que sugere que o látex é citotóxico. Uma alta incidência de aberrações cromossômicas em células tratadas com o látex de J. curcas também foi observada, indicando que o látex apresenta efeito genotóxico. Essa análise sugere que o látex de J. curcas possui efeitos tóxico, citotóxico e genotóxico, sendo que o uso indiscriminado do látex de J. curcas na medicina popular pode trazer risco à saúde humana.

Bioactive Compounds Found in Brazilian Cerrado Fruits.

Functional foods include any natural product that presents health-promoting effects, thereby reducing the risk of chronic diseases. Cerrado fruits are considered a source of bioactive substances, mainly phenolic compounds, making them important functional foods. Despite this, the losses of natural vegetation in the Cerrado are progressive. Hence, the knowledge propagation about the importance of the species found in Cerrado could contribute to the preservation of this biome. This review provides information about Cerrado fruits and highlights the structures and pharmacologic potential of functional compounds found in these fruits. Compounds detected in Caryocar brasiliense Camb. (pequi), Dipteryx alata Vog. (baru), Eugenia dysenterica DC. (cagaita), Eugenia uniflora L. (pitanga), Genipa americana L. (jenipapo), Hancornia speciosa Gomes (mangaba), Mauritia flexuosa L.f. (buriti), Myrciaria cauliflora (DC) Berg (jabuticaba), Psidium guajava L. (goiaba), Psidium spp. (araçá), Solanum lycocarpum St. Hill (lobeira), Spondias mombin L. (cajá), Annona crassiflora Mart. (araticum), among others are reported here.

Hydroxamate production as a high affinity iron acquisition mechanism in Paracoccidioides spp.

Iron is a micronutrient required by almost all living organisms, including fungi. Although this metal is abundant, its bioavailability is low either in aerobic environments or within mammalian hosts. As a consequence, pathogenic microorganisms evolved high affinity iron acquisition mechanisms which include the production and uptake of siderophores. Here we investigated the utilization of these molecules by species of the Paracoccidioides genus, the causative agents of a systemic mycosis. It was demonstrated that iron starvation induces the expression of Paracoccidioides ortholog genes for siderophore biosynthesis and transport. Reversed-phase HPLC analysis revealed that the fungus produces and secretes coprogen B, which generates dimerumic acid as a breakdown product. Ferricrocin and ferrichrome C were detected in Paracoccidioides as the intracellular produced siderophores. Moreover, the fungus is also able to grow in presence of siderophores as the only iron sources, demonstrating that beyond producing, Paracoccidioides is also able to utilize siderophores for growth, including the xenosiderophore ferrioxamine. Exposure to exogenous ferrioxamine and dimerumic acid increased fungus survival during co-cultivation with macrophages indicating that these molecules play a role during host-pathogen interaction. Furthermore, cross-feeding experiments revealed that Paracoccidioides siderophores promotes growth of Aspergillus nidulans strain unable to produce these iron chelators. Together, these data denote that synthesis and utilization of siderophores is a mechanism used by Paracoccidioides to surpass iron limitation. As iron paucity is found within the host, siderophore production may be related to fungus pathogenicity.

Hemoglobin uptake by Paracoccidioides spp. is receptor-mediated.

Iron is essential for the proliferation of fungal pathogens during infection. The availability of iron is limited due to its association with host proteins. Fungal pathogens have evolved different mechanisms to acquire iron from host; however, little is known regarding how Paracoccidioides species incorporate and metabolize this ion. In this work, host iron sources that are used by Paracoccidioides spp. were investigated. Robust fungal growth in the presence of the iron-containing molecules hemin and hemoglobin was observed. Paracoccidioides spp. present hemolytic activity and have the ability to internalize a protoporphyrin ring. Using real-time PCR and nanoUPLC-MSE proteomic approaches, fungal growth in the presence of hemoglobin was shown to result in the positive regulation of transcripts that encode putative hemoglobin receptors, in addition to the induction of proteins that are required for amino acid metabolism and vacuolar protein degradation. In fact, one hemoglobin receptor ortholog, Rbt5, was identified as a surface GPI-anchored protein that recognized hemin, protoporphyrin and hemoglobin in vitro. Antisense RNA technology and Agrobacterium tumefaciens-mediated transformation were used to generate mitotically stable Pbrbt5 mutants. The knockdown strain had a lower survival inside macrophages and in mouse spleen when compared with the parental strain, which suggested that Rbt5 could act as a virulence factor. In summary, our data indicate that Paracoccidioides spp. can use hemoglobin as an iron source most likely through receptor-mediated pathways that might be relevant for pathogenic mechanisms.

Characterization of the Paracoccidioides beta-1,3-glucanosyltransferase family.

The cell wall of pathogenic microbes acts as an initial barrier that is in contact with hostile environments. Several proteins are associated to the cell wall, including the glucanosyltransferases, which are attached through glycosylphosphatidylinositol anchors to the wall. Here, we characterized the Paracoccidioides beta-1,3-glucanosyltranferase ( Gel ) family of proteins that showed significant homology to proteins belonging to the GH72 family. Immunoassays demonstrated Gel1p associated with the cell wall and with the nucleus. For Gel2p, immune labeling was associated with the cell wall and cytoplasm. Genetic complementation studies in Saccharomyces cerevisiae demonstrated that Gel2p is able to participate in the maintenance of fungal cell wall integrity, as it was able to restore the lack of Gas1p activity in a gas1Δ mutant; Gel1p was not able to do the same. On the other hand, Gel1p restores telomeric silencing in a gas1Δ mutant, providing strong support that Gel1p can be involved in transcriptional silencing in Paracoccidioides. Use of the in vivo yeast two-hybrid system revealed proteins that interact with Paracoccidioides Gel proteins, supporting new insights into the function of Gel family members and suggesting that they could play other roles than those established at the fungal cell wall.