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Introduction: Approximately 90% of reported and identified cases of snakebites in Brazil are caused by species belonging to the Bothrops genus. These snakes have clinical relevance due to their venom composition, which contains substances capable of triggering local and systemic effects, leading to morbidities and/or mortality. Objective: The objective of this study was to evaluate the toxic and toxinological effects of Bothrops jararacussu snake venom on zebrafish embryos and larvae. Methods: The stability of B. jararacussu snake venom under the conditions used in the toxicity experiments in zebrafish embryos and larvae was evaluated on citrated human plasma. Zebrafish embryos and/or larvae mortality, morphological alterations, spontaneous tail movements and heartbeat caused by the venom were quantified within 96 hours. Toxicity parameters and activity of enzyme-related toxicity biomarkers were evaluated in zebrafish after 96 hours of semi-static exposure to the venom. Results: The results indicated that the venom causes toxicity in zebrafish embryos and larvae, inducing embryonic mortality, alteration in the number of spontaneous tail movements and activity of biomarker enzymes. The results suggested that the toxic effects caused by the venom in the early stages of zebrafish development are mediated, in part, by neurotoxic action, induction of oxidative and metabolic stress caused by low molecular weight components, and proteins present in this venom. Conclusion: Toxinological evaluations using the zebrafish as a model are scarce; however, this study presented promising results that encourage the development of future research in toxinology using this animal as a model organism.

Graphical abstract, Highlights • 25H-NBOMe and 25H-NBOH recreational drugs induces abnormal formation in zebrafish embryos. • Biophysical and theoretical studies indicate that these drugs have affinity for the DNA major groove. • The toxicity observed in the zebrafish embryos and DNA interaction may be correlated., Toxicological effects of 25H-NBOMe and 25H-NBOH recreational drugs on zebrafish embryos and larvae at the end of 96 h exposure period were demonstrated. 25H-NBOH and 25H-NBOMe caused high embryo mortality at 80 and 100 µg mL−1, respectively. According to the decrease in the concentration tested, lethality decreased while non-lethal effects were predominant up to 10 and 50 µg mL−1 of 25H-NBOH and 25H-NBOMe, respectively, including spine malformation, egg hatching delay, body malformation, otolith malformation, pericardial edema, and blood clotting. We can disclose that these drugs have an affinity for DNA in vitro using biophysical spectroscopic assays and molecular modeling methods. The experiments demonstrated that 25H-NBOH and 25H-NBOMe bind to the unclassical major groove of ctDNA with a binding constant of 27.00 × 104 M−1 and 5.27 × 104 M−1, respectively. Furthermore, these interactions lead to conformational changes in the DNA structure. Therefore, the results observed in the zebrafish embryos and DNA may be correlated.

This study evaluated the in vitro antimicrobial and immunomodulatory action of crude extracts from Anacardium occidentale L. (cashew tree) leaves and bark, and to determine their toxicity to peripheral-blood mononuclear cells (PBMCs) and to zebrafish embryos and larvae. Chemical analysis of extracts was performed by proton nuclear magnetic resonance (1H-NMR). The antibacterial activity was evaluated against selected bacteria strains by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Cytotoxicity of the extracts was assessed using resazurin method, while the effect on production of ROS by PMN leukocytes was measured by luminol. Embryotoxicity to zebrafish was assessed using the fish embryo acute toxicity test (FET) and quantification of toxicity marker enzymes (AChE, LDH, and GST). 1H-NMR results showed anacardic acid as the main component of the extracts. All bacterial species tested were sensitive to the extracts, with MICs ranging from 312.5 to 10,000 μg/mL. Streptococcus mutans and Escherichia coli were the most susceptible species. The extracts promoted cell viability above 75% at concentrations from 1.25 to 80 μg/mL. Both extracts reduced zymosan-induced ROS (p < 0.05) at concentrations of 1, 8, and 80 μg/mL compared to the control. In vivo, there were embryotoxic effects in zebrafish embryos exposed to both extracts through the presence of lethal and sublethal endpoints. The samples also acted by inhibiting the activities of biomarker enzymes. The A. occidentale L. bark and leaf extracts showed antimicrobial potential and modulated ROS production in vitro, but these also showed embryotoxic effects to zebrafish.

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The Triplaris gardneriana Wedd. seeds extract has great therapeutic potential due to numerous biological activities such as antioxidant, antibacterial and anti-inflammatory, which are associated wi...

BACKGROUND/AIM Studies with acridine compounds have reported anticancer effects. Herein, we evaluated the toxicity and antitumor effect of the (E)-1'-((4-chlorobenzylidene)amino)-5'-oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-06), a promising anticancer spiro-acridine compound. MATERIALS AND METHODS The toxicity of AMTAC-06 was evaluated on zebrafish and mice. Antitumor activity was assessed in Ehrlich ascites carcinoma model. Effects on angiogenesis, cytokine levels and cell cycle were also investigated. RESULTS AMTAC-06 did not induce toxicity on zebrafish and mice (LD50 approximately 5000 mg/kg, intraperitoneally). No genotoxicity was observed on micronucleus assay. AMTAC-06 significantly reduced the total viable Ehrlich tumor cells and increased sub-G1 peak, suggesting apoptosis was triggered. Moreover, the compound significantly decreased the density of peritumoral microvessels, indicating an anti-angiogenic action, possibly dependent on the cytokine modulation (TNF-α, IL-1β and IFN-γ). No significant toxicological effects were recorded for AMTAC-06 on tumor transplanted animals. CONCLUSION AMTAC-06 has low toxicity and a significant antitumor activity.

The use of zebrafish (Danio rerio) has arisen as a promising biological platform for toxicity testing of pesticides such as herbicides, insecticides, and fungicides. Therefore, it is relevant to assess the use of zebrafish in models of exposure to investigate the diversity of pesticide-associated toxicity endpoints which have been reported. Thus, this review aimed to assess the recent literature on the use of zebrafish in pesticide toxicity studies to capture data on the types of pesticide used, classes of pesticides, and zebrafish life stages associated with toxicity endpoints and phenotypic observations. A total of 352 articles published between September 2012 and May 2019 were curated. The results show an increased trend in the use of zebrafish for testing the toxicity of pesticides, with a great diversity of pesticides (203) and chemical classes (58) with different applications (41) being used. Furthermore, experimental outcomes could be clustered in 13 toxicity endpoints, mainly developmental toxicity, oxidative stress, and neurotoxicity. Organophosphorus, pyrethroid, azole, and triazine were the most studied classes of pesticides and associated with various toxicity endpoints. Studies frequently opted for early life stages (embryos and larvae). Although there is an evident lack of standardization of nomenclatures and phenotypic alterations, the information gathered here highlights associations between (classes of) pesticides and endpoints, which can be used to relate mechanisms of action specific to certain classes of chemicals.

The zebrafish (Danio rerio) is a small teleost fish that is becoming increasingly popular in laboratories worldwide and several attributes have also placed the zebrafish under the spotlight of (eco)toxicological studies. Since the 1990s, international organizations such as ISO and OECD have published guidelines for the use of zebrafish in ecotoxicological assessment of environmental toxicants such as the Fish Embryo Acute Toxicity (FET) test, OECD n degrees 236 guideline. This protocol uses 3,4-dichloroaniline (DCA), an aniline pesticide whose toxicity to fish species at early life stages is well known, as a positive control. Despite its use, little is known about its molecular mechanisms, especially in the context of the FET test. Therefore, this study aimed to investigate such changes in zebrafish larvae exposed to DCA (4 mg/L) for 96 hours using gel-free proteomics. Twenty-four proteins detected in both groups were identified as significantly affected by DCA exposure, and, when considering group-specific entities, 48 proteins were exclusive to DCA (group-specific proteins) while 248 were only detected in the control group. Proteins modulated by DCA treatment were found to be involved in metabolic processes, especially lipids and hormone metabolism (eg, Apoa1 and Apoa1b and vitelogenins), as well as proteins important for developmental processes and organogenesis (eg, Myhc4, Acta2, Sncb, and Marcksb). The results presented here may therefore provide a better understanding of the relationships between molecular changes and phenotype in zebrafish larvae treated with DCA, the reference compound of the FET test.

The use of unproven COVID‐19 drugs has seen a massive increase in recent months in Brazil. Chloroquine (CQ), hydroxychloroquine (HCQ) and ivermectin (IVM) are among the most politically promoted and sold drugs. As a result, an increased demand for these pharmaceuticals has been observed across the country with sales increase by more than 50% in comparison to 2019. In a country where only 46% of domestic wastewater is treated, what can we expect from the effects of these drugs and their metabolites on aquatic wildlife? The environmental fate and ecotoxicity data for IVM indicate varying degrees of toxicity to aquatic invertebrates and the potential to accumulate in several aquatic organisms. The CQ compound was classified as harmful to aquatic organisms and may cause long‐term adverse effects in the aquatic environment. As for HCQ, it has been tagged as posing insignificant environmental risks in the pre‐pandemic scenario despite its environmental persistence. Fragile Brazilian biomes, such as the Amazon (North region), Caatinga and coastal region of the Northeast and the Atlantic Rainforest (Southeast region) are of particular concern given the high incidence of COVID‐19 cases in these areas. The large‐scale use of unproven drugs in response to the pandemic may add undue weight to the increased volume of pharmaceuticals entering the environment, leaving the local biota vulnerable to the harmful effects of these contaminants. A nationwide coordinated response is urgently needed to monitor the levels and investigate the impacts of these emerging contaminants. This article is protected by copyright. All rights reserved.

A tutoria faz parte dos programas acadêmicos oferecidos pela Universidade Federal da Paraíba (UFPB), com objetivo de auxiliar os discentes a partir da implementação de estratégias que facilitem o percurso dos estudantes pelas disciplinas com maiores índices de reprovação e evasão. Este trabalho teve como objetivo avaliar o papel do tutor como agente no processo de ensino e aprendizagem na disciplina de Bioquímica Metabólica, na UFPB. Para tanto, os tutores dessa disciplina aplicaram quatro estudos dirigidos e um jogo didático nas turmas dos períodos letivos 2016.2 (Grupo 1/n= 27) e 2017.1 (Grupo 2/n= 23). O desempenho dos discentes que usufruíram dos recursos didáticos foi satisfatório e significativamente maior do que o grupo que não participou. A partir da avaliação da tutoria pelos discentes por meio de um questionário online, respondeu-se unanimemente que a disciplina de Bioquímica Metabólica precisa ofertar tutores, declarando que os mesmos são facilitadores importantes no processo de ensino e aprendizagem.

A tutoria faz parte dos programas acadêmicos oferecidos pela Universidade Federal da Paraíba (UFPB), com objetivo de auxiliar os discentes a partir da implementação de estratégias que facilitem o percurso dos estudantes pelas disciplinas com maiores índices de reprovação e evasão. Este trabalho teve como objetivo avaliar o papel do tutor como agente no processo de ensino e aprendizagem na disciplina de Bioquímica Metabólica, na UFPB. Para tanto, os tutores dessa disciplina aplicaram quatro estudos dirigidos e um jogo didático nas turmas dos períodos letivos 2016.2 (Grupo 1/n= 27) e 2017.1 (Grupo 2/n= 23). O desempenho dos discentes que usufruíram dos recursos didáticos foi satisfatório e significativamente maior do que o grupo que não participou. A partir da avaliação da tutoria pelos discentes por meio de um questionário online, respondeu-se unanimemente que a disciplina de Bioquímica Metabólica precisa ofertar tutores, declarando que os mesmos são facilitadores importantes no processo de ensino e aprendizagem. The tutoring is part of the academic programs offered by the Federal University of Paraíba (UFPB), aiming to help students from the implementation of strategies that facilitate the students' journey through the disciplines with the highest rates of retention and dropout. This work aimed to evaluate the role of the tutor as an agent in the teaching and learning process in the discipline of Metabolic Biochemistry, at UFPB. To that end, the tutors of this discipline applied four directed studies and a didactic game in the classes of 2016.2 (Group 1/n= 27) and 2017.1 (Group 2/n= 23) periods. The performance of the students who were benefited from the didactic resources was satisfactory and significantly higher than the group that did not participate. From the evaluation of the tutoring by the students through an online questionnaire, it was unanimous that the discipline of Metabolic Biochemistry must provide tutors, stating that they are important supporters in the teaching and learning process.

The tutoring is part of the academic programs offered by the Federal University of Paraíba (UFPB), aiming to help students from the implementation of strategies that facilitate the students' journey through the disciplines with the highest rates of retention and dropout. This work aimed to evaluate the role of the tutor as an agent in the teaching and learning process in the discipline of Metabolic Biochemistry, at UFPB. To that end, the tutors of this discipline applied four directed studies and a didactic game in the classes of 2016.2 (Group 1/n= 27) and 2017.1 (Group 2/n= 23) periods. The performance of the students who were benefited from the didactic resources was satisfactory and significantly higher than the group that did not participate. From the evaluation of the tutoring by the students through an online questionnaire, it was unanimous that the discipline of Metabolic Biochemistry must provide tutors, stating that they are important supporters in the teaching and learning process.

Cry1C, Cry1F and Cry1Ab are insecticidal proteins from Bacillus thuringiensis (Bt) which are expressed in transgenic crops. Given the entry of these proteins into aquatic environments, it is relevant to evaluate their impacts on aquatic organisms. In this work, we sought to evaluate the effects of Cry1C, Cry1F and Cry1Ab on zebrafish embryos and larvae of a predicted worst-case scenario concentration of these proteins (set to 1.1 mg/L). For that, we coupled a traditional toxicity approach (the zebrafish embryotoxicity test and dosage of enzymatic biomarkers) to gel free proteomics analysis. At the concentration tested, these proteins did not cause adverse effects in the zebrafish early life stages, either by verifying phenotypic endpoints of toxicity or alterations in representative enzymatic biomarkers (catalase, glutathione-S-tranferase and lactate-dehydrogenase). At the molecular level, the Cry proteins tested lead to very small changes in the proteome of zebrafish larvae. In a global way, these proteins upregulated the expression of vitellogenins. Besides that, Cry1C e Cry1F deregulated heterogeneous nuclear ribonucleoproteins (Hnrnpa0l and Hnrnpaba, respectively), implicated in mRNA processing and gene regulation. Overall, these data lead to the conclusion that Cry1C, Cry1F and Cry1Ab proteins, even at a very high concentration, have limited effects in the early stages of zebrafish life. (C) 2020 Elsevier Ltd. All rights reserved.

The increasing use and disposal of plastics has become a persistent problem in the marine environment, calling for studies that refer to realistic scenarios to understand their effects on biota. Particularly, the understanding about the effects of co-exposure with nanoplastic particles and metals on aquatic organisms is still limited. The present work aimed to investigate the acute toxicity of amino-functionalized polystyrene nanoparticles (PS-NH2; 50 nm) as proxy for nanoplastics on brine shrimp Artemia franciscana larvae under different culture conditions and at different stages of development, as well as the combined effect with two reference toxicants - potassium dichromate (K2Cr2O7) and copper sulfate (CuSO4). Nauplii (instar II or III larval stages) were exposed to different concentrations of PS-NH2 (0.005 to 5 μg mL-1) for up to 48 h, with or without agitation in order to mimic a more realistic environmental scenario. Larval mobility and PS-NH2 accumulation were monitored under microscopy. PS-NH2 alone showed toxicity only at the highest concentration tested (5 μg mL-1) regardless the incubation method used (61.2 + 3.1% and 65.0 + 4.5% with and without agitation, respectively). Moreover, instar III stage was the most sensitive to PS-NH2 exposure (38.2% immobility in 24 h of exposure; 5 μg mL-1). Evidence of PS-NH2 retention in the gastrointestinal tract in a concentration- and time-dependent manner was also obtained. Mixtures of PS-NH2 (0.005 and 5 μg mL-1) with different concentrations of K2Cr2O7 increased the immobilization rate of the larvae after 48 h of exposure, when compared to the K2Cr2O7 alone. Similar results were observed for CuSO4 in the co-exposure conditions at different concentrations. However, exposing nauplii to a mixture of PS-NH2 (0.005 μg mL-1) and CuSO4 decreased immobilization rate, in comparison to the group exposed to CuSO4 alone. The present work highlights the potential risk posed by nanoplastics to zooplanktonic species through their interaction with other toxicants.

Mimosa caesalpiniifolia (Fabaceae) is used by Brazilian people to treat hypertension, bronchitis, and skin infections. Herein, we evaluated the antiproliferative action of the dichloromethane fraction from M. caesalpiniifolia (DFMC) stem bark on murine tumor cells and the in vivo toxicogenetic profile. Initially, the cytotoxic activity of DFMC on primary cultures of Sarcoma 180 (S180) cells by Alamar Blue, trypan, and cytokinesis block micronucleus (CBMN) assays was assessed after 72 h of exposure, followed by the treatment of S180-bearing Swiss mice for 7 days, physiological investigations, and DNA/chromosomal damage. DFMC and betulinic acid revealed similar in vitro antiproliferative action on S180 cells and induced a reduction in viable cells, induced a reduction in viable cells and caused the emergence of bridges, buds, and morphological features of apoptosis and necrosis. S180-transplanted mice treated with DFMC (50 and 100 mg/kg/day), a betulinic acid-rich dichloromethane, showed for the first time in vivo tumor growth reduction (64.8 and 80.0%) and poorer peri- and intratumor quantities of vessels. Such antiproliferative action was associated with detectible side effects (loss of weight, reduction of spleen, lymphocytopenia, and neutrophilia and increasing of GOT and micronucleus in bone marrow), but preclinical general anticancer properties of the DFMC were not threatened by toxicological effects, and these biomedical discoveries validate the ethnopharmacological reputation of Mimosa species as emerging phytotherapy sources of lead molecules.

Objective. We aimed to define the safety and toxicity of both isolated and embedded cinnamaldehyde using a pharmaceutical formulation for the treatment of oral fungal infections in an in vivo study. Materials and Methods. Acute toxicity was assessed in studies with Galleria mellonella larvae and Danio rerio embryos (zebrafish), and genotoxicity was assessed in a mouse model. The pharmaceutical formulation (orabase ointment) containing cinnamaldehyde was evaluated for verification of both in vitro antifungal activity and toxicity in keratinized oral rat mucosa. Results. In Galleria mellonella larvae, cinnamaldehyde was not toxic up to the highest dose tested (20 mg/kg) and presented no genotoxicity up to the dose of 4 mg/kg in the model using mice. However, it was found to be toxic in zebrafish embryos up to a concentration of 0.035 μg/mL; LC50 0.311; EC50 0.097 (egg hatching delay); and 0.105 (Pericardial edema). In the orabase antifungal susceptibility test, cinnamaldehyde exhibited activity in concentrations greater than 200 μg/mL. As for safety in the animal model with rats, the orabase ointment proved to be safe for use on keratinized mucosa up to the maximum concentration tested (700 μg/mL). Conclusions. At the concentrations tested, cinnamaldehyde was not toxic in vertebrate and invertebrate animal models and did not exhibit genotoxic activity. In addition, when used in the form of an ointment in orabase, having already recognized antifungal activity, it was shown to be safe up to the highest concentration tested.

Moxidectin is an antiparasitic drug belonging to the class of the macrocyclic lactones, subgroup mylbemicins. It is used worldwide in veterinary practice, but little is known about its potential environmental risks. Thus, we used the zebrafish embryo as a model system to study the potential effects of moxidectin on aquatic non-target organisms. The analyses were performed in two experimental sets: (1) acute toxicity and apical endpoints were characterized, with biomarker assays providing information on the activity levels of catalase (CAT), glutathione S-transferase (GST), lactate dehydrogenase (LDH), and acetylcholinesterase (AChE); and (2) internal concentration and spatial distribution of moxidectin were determined using ultraperformance liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-QToF-MS) and matrix-assisted laser desorption/ionization-MS imaging (MALDI-MSi). The acute toxicity to zebrafish embryos (96 hpf) appeared mainly as a decrease in hatching rates (EC50 = 20.75 μg/L). It also altered the enzymatic activity of biomarker enzymes related to xenobiotic processing, anaerobic metabolism, and oxidative stress (GST, LDH, and CAT, respectively) and strongly accumulated in the embryos, as internal concentrations were 4 orders of magnitude higher than those detected in exposure solutions. MALDI-MSi revealed accumulations of the drug mainly in the head and eyes of the embryos (72 and 96 hpf). Thus, our results show that exposure to moxidectin decreases hatching success by 96 h and alters biochemical parameters in the early life stages of zebrafish while accumulating in the head and eye regions of the animals, demonstrating the need to prioritize this compound for environmental studies.

Triplaris gardneriana Wedd. is a tree used in folk medicine to treat venereal diseases and inflammation as well as a source of biological compounds with antioxidant capacity. In order to assess the safety of these bioactive compounds, the present study aimed to determine the toxicity of an ethanolic extract of T. gardneriana, (EETg). Toxicological tests included hemolytic activity, toxicity toward the brine shrimp Artemia, cytotoxicity against breast cancer cells (MCF7) and acute oral toxicity in rodents. In addition, toxicogenomics techniques were used to determine genome expression in MCF7 cells exposed to EETg. The results showed that the extract exhibits approximately 60% of hemolytic activity at the highest tested concentration (64 mu g/ml) and toxicity against nauplii of Artemia sp. (LC50 of 67.85 mu g/ml). Further, EETg appears to be cytotoxic to MCF7 (cell viability reduced to 40% at 250 mu g/ml after 24 hr). Genomic data demonstrated differential expression of 14 genes. Data analysis indicated possible altered pathways (e.g., xenobiotic metabolism), possible adverse health risks (e.g., hepatotoxicity), and drugs with similar gene expression profile (e.g., antimicrobials). The investigation provides important information on potentially adverse aspects of EETg, which need to be considered prior to the therapeutic utilization of this plant.

Jaburetox (JBTX) is an insecticidal and antifungal peptide derived from jack bean (Canavalia ensiformis) urease that has been considered a candidate for developing genetically modified crops. This study aimed to perform the risk assessment of the peptide JBTX following the general recommendations of the two-tiered, weight-of-evidence approach proposed by International Life Sciences Institute. The urease of C. ensiformis (JBU) and its isoform JBURE IIb (the JBTX parental protein) were assessed. The history of safe use revealed no hazard reports for the studied proteins. The available information shows that JBTX possesses selective activity against insects and fungi. JBTX and JBU primary amino acids sequences showed no relevant similarity to toxic, antinutritional or allergenic proteins. Additionally, JBTX and JBU were susceptible to in vitro digestibility, and JBU was also susceptible to heat treatment. The results did not identify potential risks of adverse effects and reactions associated to JBTX. However, further allergen (e.g. serum IgE binding test) and toxicity (e.g. rodent toxicity tests) experimentation can be done to gather additional safety information on JBTX, and to meet regulatory inquiries for commercial approval of transgenic cultivars expressing this peptide.

Soybean toxin (SBTX) is a protein isolated from soybean seeds and composed of two polypeptide subunits (17 and 27 kDa). SBTX has in vitro activity against phytopathogenic fungi such as Cercospora sojina, Aspergillus niger, and Penicillium herguei, and yeasts like Candida albicans, C. parapsilosis, Kluyveromyces marxiannus, and Pichia membranifaciens. The present study aimed to analyze in vitro whether SBTX causes any side effects on non-target bacterial and mammalian cells that could impede its potential use as a novel antifungal agent. SBTX at 100 mu g/mL and 200 mu g/mL did not hinder the growth of the bacteria Salmonella enterica (subspecies enterica serovar choleraesuis), Bacillus subtilis (subspecies spizizenii) and Staphylococcus aureus. Moreover, SBTX at concentrations up to 500 mu g/mL did not significantly affect the viability of erythrocytes, neutrophils, and human intestinal Caco-2 cells. To study whether SBTX could induce relevant alterations in gene expression, in vitro DNA microarray experiments were conducted in which differentiated Caco-2 cells were exposed for 24 h to 100 mu g/mL or 200 mu g/mL SBTX. SBTX up-regulated genes involved in cell cycle and immune response pathways, but downregulated genes that play a role in cholesterol biosynthesis and platelet degranulation pathways. Thus, although SBTX did not affect bacteria, nor induced cytotoxity in mammalian cells, it affected some biological pathways in the human Caco-2 cell line that warrants further investigation.

2,4-Dichlorophenoxyacetic acid (2,4-D) herbicide is the main ingredient in over 1500 commercially available products such as Weedestroy® AM40 and DMA® 4 IVM. Although the liver has been identified as one of the organs that are affected by this herbicide, reports on its hepatotoxic effects available in the literature are restricted to rats. Thus, there is a gap in information on other organisms that may be vulnerable to 2,4-D exposure, such as fish. Therefore, the present work aimed to assess the hepatotoxic potential of 2,4-D in fish using zebrafish (Danio rerio) larvae as a model system. For this purpose, its acute toxicity to zebrafish embryos was assessed, as well as its sublethal effects (

The antitumor effects of thiophene and acridine compounds have been described; however, the clinical usefulness of these compounds is limited due to the risk of high toxicity and drug resistance. The strategy of molecular hybridization presents the opportunity to develop new drugs which may display better target affinity and less serious side effects. Herein, 2-((6-Chloro-2-methoxy-acridin-9-yl)amino)-5,6,7,8-tetrahydro-4H-cyclohepta[b]-thiophene-3-carbonitrile (ACS03), a hybrid thiophene–acridine compound with antileishmanial activity, was tested for toxicity and antitumor activity. The toxicity was evaluated in vitro (on HaCat and peripheral blood mononuclear cells) and in vivo (zebrafish embryos and acute toxicity in mice). Antitumor activity was also assessed in vitro in HCT-116 (human colon carcinoma cell line), K562 (chronic myeloid leukemic cell line), HL-60 (human promyelocytic leukemia cell line), HeLa (human cervical cancer cell line), and MCF-7 (breast cancer cell line) and in vivo (Ehrlich ascites carcinoma model). ACS03 exhibited selectivity toward HCT-116 cells (Half maximal inhibitory concentration, IC50 = 23.11 ± 1.03 µM). In zebrafish embryos, ACS03 induced an increase in lactate dehydrogenase, glutathione S-transferase, and acetylcholinesterase activities. The LD50 (lethal dose 50%) value in mice was estimated to be higher than 5000 mg/kg (intraperitoneally). In vivo, ACS03 (12.5 mg/kg) induced a significant reduction in tumor volume and cell viability. In vivo antitumor activity was associated with the nitric oxide cytotoxic effect. In conclusion, significant antitumor activity and weak toxicity were recorded for this hybrid compound, characterizing it as a potential anticancer compound.

Structural diversity characterizes natural products as prototypes for design of lead compounds. The aim of this study was to synthetize, and to evaluate the toxicity and antitumor action of a new piperine analogue, the butyl 4-(4-nitrobenzoate)-piperinoate (DE-07). Toxicity was evaluated against zebrafish, and in mice (acute and micronucleus assays). To evaluate the DE-07 antitumor activity Ehrlich ascites carcinoma model was used in mice. Angiogenesis, Reactive Oxygen Species (ROS) production and cytokines levels were investigated. Ninety-six hours exposure to DE-07 did not cause morphological or developmental changes in zebrafish embryos and larvae, with estimated LC50 (lethal concentration 50%) higher than 100 μg/mL. On the acute toxicity assay in mice, LD50 (lethal dose 50%) was estimated at around 1000 mg/kg, intraperitoneally (i.p.). DE-07 (300 mg/kg, i.p.) did not induce increase in the number of micronucleated erythrocytes in mice, suggesting no genotoxicity. On Ehrlich tumor model, DE-07 (12.5, 25 or 50 mg/kg, i.p.) induced a significant decrease on cell viability. In addition, there was an increase on ROS production and a decrease in peritumoral microvessels density. Moreover, DE-07 induced an increase of cytokines levels involved in oxidative stress and antiangiogenic effect (IL-1β, TNF-α and IL-4). No significant clinical toxicological effects were recorded in Ehrlich tumor transplanted animals. These data provide evidence that DE-07 presents low toxicity, and antitumor effect via oxidative and antiangiogenic actions by inducing modulation of inflammatory response in the tumor microenvironment.

Licania tomentosa is a Brazilian plant species that produces edible fruits, yet there is little information available concerning their nutritional and/or bioactive composition. This study aimed to evaluate the nutritional and polyphenol composition of L. tomentosa fruits (pulp and seeds) and measure antioxidant activity in ethanolic extracts.The pulp and seeds were excellent sources of fiber (25.62%–41.70%) as well as minerals and vitamins. L. tomentosa contained no lectins or protease inhibitors (chymotrysin and trypsin) and 12 polyphenol compounds were identified in the seed extracts with a predominance of flavonoids. The seeds also presented antioxidant activities using the DPPH (SC5010.30–15.87 μg/mL), TBARS (IC50 18.46–20.84 μg/mL), and FRAP (RC50 0.203–0.309 μg/mL) assays. Due to its nutrient and antioxidant content, L. tomentosa may be used for food applications.

Cysteine peptidases (EC 3.4.22) are the most abundant enzymes in latex fluids. However, their physiological functions are still poorly understood, mainly related to defense against phytopathogens. The present study reports the cDNA cloning and sequencing of five undescribed cysteine peptidases from Calotropis procera (Aiton) Dryand (Apocynaceae) as well as some in silico analyses. Of these, three cysteine peptidases (CpCP1, CpCP2, and CpCP3) were purified. Their enzymatic kinetics were determined and they were assayed for their efficacy in inhibiting the hyphal growth of phytopathogenic fungi. The mechanism of action was investigated by fluorescence and atomic force microscopy as well as by induction of reactive oxygen species (ROS). The deduced amino acid sequences showed similar biochemical characteristics and high sequence homology with several other papain-like cysteine peptidases. Three-dimensional models showed two typical cysteine peptidase domains (L and R domains), forming a “V-shaped” active site containing the catalytic triad (Cys, His, and Asn). Proteolysis of CpCP1 was higher at pH 7.0, whereas for CpCP2 and CpCP3 it was higher at 7.5. All peptidases exhibited optimum activity at 35 °C and followed Michaelis-Menten kinetics. However, the major difference among them was that CpCP1 exhibited highest Vmax, Km, Kcat and catalytic efficiency. All peptidases were deleterious to the two fungi tested, with IC50 of around 50 μg/mL. The peptidases promoted membrane permeabilization, morphological changes with leakage of cellular content, and induction of ROS in F. oxysporum spores. These results corroborate the hypothesis that latex cysteine peptidases play a role in defense against fungi.

The biochemical and nutritional attributes of two soybean (Glycine max (L.) Merr.) cultivars, one susceptible (Seridó) and the other resistant (Seridó-RCH) to stem canker, were examined to assess whether the resistance to pathogens was related to levels of antinutritional and/or defense proteins in the plant and subsequently affected the nutritional quality. Lectin, urease, trypsin inhibitor, peroxidase and chitinase activities were higher in the resistant cultivar. Growing rats were fed with isocaloric and isoproteic diets prepared with defatted raw soybean meals. Those on the Seridó-RCH diet showed the worst performance in terms of protein quality indicators. Based on regression analysis, lectin, trypsin inhibitor, peroxidase and chitinase appear to be involved in the resistance trait but also in the poorer nutritional quality of Seridó-RCH. Thus, the development of cultivars for disease resistance may lead to higher concentrations of antinutritional compounds, affecting the quality of soybean seeds. Further research that includes the assessment of more cultivars/genotypes is needed.

Moringa oleifera has been used in traditional medicine to treat diabetes. However, few studies have been conducted to relate its antidiabetic properties to proteins. In this study, a leaf protein isolate was obtained from M. oleifera leaves, named Mo-LPI, and the hypoglycemic and antioxidant effects on alloxan-induced diabetic mice were assessed. Mo-LPI was obtained by aqueous extraction, ammonium sulphate precipitation and dialysis. The electrophoresis profile and proteolytic hydrolysis confirmed its protein nature. Mo-LPI showed hemagglutinating activity, cross-reaction with anti-insulin antibodies and precipitation after zinc addition. Single-dose intraperitoneal (i.p.) administration of Mo-LPI (500 mg/kg·bw) reduced the blood glucose level (reductions of 34.3%, 60.9% and 66.4% after 1, 3 and 5 h, respectively). The effect of Mo-LPI was also evidenced in the repeated dose test with a 56.2% reduction in the blood glucose level on the 7th day after i.p. administration. Mo-LPI did not stimulate insulin secretion in diabetic mice. Mo-LPI was also effective in reducing the oxidative stress in diabetic mice by a decrease in malondialdehyde level and increase in catalase activity. Mo-LPI (2500 mg/kg·bw) did not cause acute toxicity to mice. Mo-LPI is a promising alternative or complementary agent to treat diabetes.

This work aimed to add value to an underexploited plant species from Brazil, Triplaris gardneriana. To that, the phenolic compounds profile of its seed ethanolic extract and fractions was examined by HPLC and the antioxidant capacity assessed using chemical assays as well as in vitro cell imaging. Twelve compounds were quantified and classified as either phenolic acids or flavonoids. The fractionation process did not generate fractions with different compositions except for chloroformic fraction, which showed only 6 out of 12 standard compounds used. DPPH assay revealed samples with a concentration-dependent radical scavenging activity, being methanolic fraction the one with the largest activity (SC50 11.45 ± 0.02 μg/mL). Lipid peroxidation assessment, in the presence and absence of stress inducer, showed that particularly the ethanol extract (IC50 26.75 ± 0.08 μg/mL) and the ethyl acetate fraction (IC50 6.14 ± 0.03 μg/mL) could inhibit lipid peroxidation. The ethyl acetate fraction performed best in chelating iron (48% complexation at 1000 μg/mL). Cell imaging experiments showed that the ethanolic extract could protect cells against oxidative stress as well as restore the oxidative balance upon stress induction. In conclusion, T. gardneriana seeds showed a promising phenolic compounds profile and antioxidant activity that may be further exploited.

This study assessed new insecticidal activities of essential oils from Lippia sidoides and Croton species (Croton zehntneri, Croton nepetaefolius, Croton argyrophylloides, and Croton sonderianus) against Aedes aegypti mosquito. In addition, the acute toxicity upon mice was determined. All essential oils showed inhibition of egg hatching, with IC50 values ranging from 66.4 to 143.2 μg mL(-1), larvicidal activity with LC50 ranging from 25.5 to 94.6 μg mL(-1), and pupicidal action with PC50 ranging from 276.8 to over 500 μg mL(-1). Only L. sidoides, C. zehntneri, and C. argyrophylloides essential oils were able to inhibit the oviposition of female gravid mosquitoes with OD50 values of 35.3, 45.3, and 45.8 μg mL(-1), respectively. Oral acute toxicity in mice showed that C. sonderianus and C. argyrophylloides oils are nontoxic (LD50 6,000 mg.kg(-1)) while C. nepetaefolius, C. zehntneri, and L. sidoides oils are moderately toxic (LD50 3,840; 3,464, and 2,624 mg.kg(-1), respectively). The results indicate that these oils are promising sources of bioactive compounds, showing low or no toxicity to mammals.

Among theBauhiniaspecies,B. cheilanthastands out for its seed protein content. However, there is no record of its nutritional value, being used in a nonsustainable way in the folk medicine and for large-scale extraction of timber. The aim of this study was to investigate the food potential ofB. cheilanthaseeds with emphasis on its protein quality to provide support for flora conservation and use as raw material or as prototype for the development of bioproducts with high added socioeconomic value.B. cheilanthaseeds show high protein content (35.9%), reasonable essential amino acids profile, low levels of antinutritional compounds, and nutritional parameters comparable to those of legumes widely used such as soybean and cowpea. The heat treatment of the seeds as well as the protein extraction process (to obtain the protein concentrate) increased the acceptance of diets by about 100% when compared to that of rawBcdiet. These wild legume seeds can be promising alternative source of food to overcome the malnutrition problem faced by low income people adding socioeconomic value to the species.

Endophytic microorganisms represent promising alternatives for obtaining new drugs of biotechnological importance. In this study, the endophytic species Acremonium cavaraeanum (A1a) isolated from Cocos nucifera was cultivated for the production of secondary metabolites, and its extracts and fractions were evaluated by the dilution method (MIC). The EtOAc extracts and MeOH fractions were tested against Gram-positive and -negative bacteria, and had an MIC of 125 µg/mL when evaluated in the EtOAc extract (EBI). The EtOAc extract (EBII) had an MIC of 62.25 µg/mL for Staphylococcus aureus and an MIC between 125 and 250 µg/mL for Gram-negative bacteria. The methanolic fractions showed activity with MIC between 125 and 250 µg/mL for all bacteria tested. The IGS region of the rDNA repeat unit of genomic DNA was analyzed by PCR/RFLPs, including endonucleases PstI, BamHII, HinfI, and EcoRI. The physical maps showed different restriction sites for the 6 Acremonium sp isolates, and revealed 5 RFLP patterns. The results showed that isolates of the same Acremonium species exhibited variation in this specific region. The sequences of ITS1-5.8S-ITS2 regions were aligned by Clustal W using the neighbor joining method, which grouped the isolates into 5 distinct clusters. This study aimed to evaluate the genetic diversity of A. cavaraeanum crops exhibiting antibacterial activity. The results of this study indicate that different fungal genetic isolates have biotechnological potential for the production of active bio-compounds against several human pathogens.

The most studied bioactive potential of phenolic compounds corresponds to antioxidant activity, which in turn, is associated with a reduction in the incidence of various human diseases. However, the total quantity of these bioactive substances in foods and medicinal preparations does not reflect the amount absorbed and metabolized by the body. The present study aimed to investigate the bioaccessibility of Triplaris gardneriana seeds ethanolic extract (EETg) by determination of phenolic composition and antioxidant activities before and after in vitro digestion as well as to estimate its bioavailability by chemical analysis of plasma and urine in animal models after oral administration. The bioaccessibility indexes of phenolic compounds in EETg were 48.65 and 69.28% in the presence and absence of enzymes, respectively. Among the identified phenolics classes, flavonoids, represented by galloylated procyanidins type B, proved to be more bioaccessible, 81.48 and 96.29% in the post-intestinal phase with and without enzymes, respectively. The oral administration in Wistar rats resulted in a significant decrease in plasma of the total antioxidant capacity, TAC, by FRAP assay 4h after beginning the experiment. For urine samples, an increase in TAC by DPPH and FRAP was observed from 1 and 4h after administration, respectively. UPLC-QTOF analysis of urine detected 2 metabolites originated from the degradation of phenolic compounds, i.e. hippuric acid and phenylacetil glycine. These results suggest that phenolic compounds in T. gardneriana are unstable under gastrointestinal conditions, being flavonoids the components with higher bioaccessibility; besides that, they showed limited bioavailability due to their rapid biotransformation and urinary elimination.

Studies have shown the benefit of antioxidants in the prevention or treatment of human diseases and promoted a growing interest in new sources of plant antioxidants for pharmacological use. This study aimed to add value to two underexploited wild plant species (Licania rigida) and L. tomentosa) from Brazilian flora. Thus, the phenolic compounds profile of their seed ethanol extract and derived fractions were elucidated by HPLC, the antioxidant capacity was assessed by in vitro chemical tests and the cytotoxicity determined using the human carcinoma cell lines MCF-7 and Caco-2. Eleven phenolic compounds were identified in the extracts of each species. The extracts and fractions showed excellent antioxidant activity in the DPPH assay (SC50, ranging from 9.15 to 248.8 μg/mL). The aqueous fraction of L. rigida seeds was most effective in preventing lipid peroxidation under basal conditions (IC50 60.80 μg/mL) whereas, in the presence of stress inducer, the methanolic fraction of L. tomentosa performed best (IC50 8.55 μg/mL). None of the samples showed iron chelating capacity. Ethanolic seed extracts of both species did not reveal any cytotoxicity against MCF-7 and Caco-2 cells. Both plant species showed a promising phenolic profile with potent antioxidant capacity and deserve attention to be sustainably explored.

BACKGROUND: Myracrodruon urundeuva Fr. Allemao is a common tree in the Caatinga that has been widely used for various medical purposes. Previous studies showed that the ethanol seed extract of M. urundeuva has potent activity against the larval stage of the dengue vector Aedes aegypti. Given this potential insecticidal activity, bioguided separation steps were performed in order to isolate the active compound(s). RESULTS: The isolation process resulted in only one active chemical compound, identified by infrared spectroscopy and mass spectrometry as m-pentadecadienyl-phenol. This compound presented potent larvicidal and pupicidal activity (LC50 10.16 and 99.06 µg mL−1 respectively) and great egg hatching inhibitory activity (IC50 49.79 µg mL−1). The mode of action was investigated through observations of behavioural and morphological changes performed in third-instar larvae treated with m-pentadecadienyl-phenol solution after 1, 6, 12, 16 and 20 h of exposure. Some changes were observed as flooding of the tracheal system, alterations in siphonal valves and anal gills and lethargy, probably caused by the strong anticholinesterasic activity reported previously. CONCLUSION: The compound isolated from M. urundeuva seeds, m-pentadecadienyl-phenol, showed potent activity against immature stages of dengue vector, Ae. aegypti, being considered the main larvicidal principle. Copyright © 2012 Society of Chemical Industry

A B S T R A C T This study aimed to establish a nutritional ranking of 30 genotypes of cowpea (Vigna unguiculata L. Walp). The results showed the proximate composition varies among genotypes in protein (20–30%) and dietary fiber contents (20–35%), in protease inhibitors (2–4 UI/mg protein), lectin (40,000–640,000 UH/kg meal) and essential amino acid levels, in vitro protein digestibility (30–40%) and in the apparent concentration of expressed proteins. The antioxidant capacity varied from EC50 of 9.54–38.7 mg seed extract/mL DPPH and the highest values detected for alpha- and delta-tocopherol were 0.38 mg/g and 1.88 mg/g, respectively. The analysis of a nutritional quality index based on the weighted average content of total protein, dietary fiber, iron, zinc, protease inhibitors and lectins allowed ranking genotypes. In descending order the 5 best genotypes were: BRS-Cauame ´ > BRS-Tumucumaque > Canapuzinho > BRS-Potengi > BRS-Urubuquara.

Seed ethanolic extracts of 21 Brazilian plants were evaluated for ovicidal, larvicidal, and pupicidal activities against insecticide-susceptible (SS) and field-collected (FC) strains of Aedes aegypti (L.) (Diptera: Culicidae), as well as for their effects on nontarget organisms. Myracrodruon urundeuva Fr. Allemao extract was highly toxic to both mosquito strains. Schinopsis brasiliensis Engler extract showed low toxicity and was 38-68 times less toxic to Ae. aegypti larvae than was M. urundeuva extract. The pupicidal activity (LC50) of 14 plant seed extracts ranged between 9 and 433/g/ml, and toxicities were comparable to both mosquito strains. Piptadenia moniliformis Benth. and Luetzelburgia auriculata (Allemao) Ducke extracts showed the highest activities against pupae of FC and SS strains. None of the extracts showed 100% ovicidal activity. In addition, the active extracts did not show high acute toxicity to mice (LD50 > 1.5 g/kg), except that of Enterolobium contortisiliquum (Vell.) Morong. Most of the active extracts exhibited low toxicity against brine shrimp (Artemia sp.) nauplii. The extracts of M. urundeuva, P. moniliformis, and L. auriculata are promising sources of recognized classes of insecticidal compounds with good selectivity against immature stages of Ae. aegypti.

The seeds of 14 wild legume species from Brazil were analyzed to assess their potential as food. The seeds showed high levels of crude protein (10.9–50.0%), dietary fiber (0.8–52.3%) and energy (905–1804 kJ 100 g−1), and the amino acid composition (mg g−1 N) was comparable to that of soybeans, but with higher amounts of lysine (456–10,884) and histidine (199–918). The seeds showed lectin activity (20–2560 and 60–2560 HU g−1 flour for untreated or treated erythrocytes, respectively) as well as trypsin inhibitory activity (4.06–27.35 μg TI mg−1 flour), urease (225–23,895 U kg−1 flour) and toxic activities (LD50 0.22–0.12 g kg−1 body weight). Most of the minerals, as with edible legume seeds, were present in high levels when considering reference daily intake (RDI) and the nutrient content claims of the United States Food and Drug Administration (FDA). Thus, these wild legume seeds can be promising alternatives sources of food to overcome malnutrition problems.

This study assessed the toxicity of seed water extracts of 15 leguminous species upon Aedes aegypti larvae. A partial chemical and biochemical characterization of water extracts, as well as the assessment of their acute toxicity in mice, were performed. The extracts of Amburana cearensis, Anadenanthera macrocarpa, Dioclea megacarpa, Enterolobium contortisiliquum and Piptadenia moniliformis caused 100% of mortalit y after 1 to 3 h of exposure. They showed LC50 and LC90 values ranging from 0.43 ± 0.01 to 9.06 ± 0.12 mg/mL and from 0.71 ± 0.02 to 13.03 ± 0.15 mg/mL, respectively. Among the secondary metabolite constituents, the seed water extracts showed tannins, phenols, flavones, favonols, xanthones, saponins and alkaloids. The extracts also showed high soluble proteins content (0.98 to 7.71 mg/mL), lectin (32 to 256 HU/mL) and trypsin inhibitory activity (3.64 = 0.43 to 26.19 = 0.05 gIT/kg of flour) The electrophoretic profiles showed a great diversity of protein bands, many of which already described as insecticide proteins. The extracts showed low toxicity to mice (LD50 > 0.15 = 0.01 g/kg body weight), but despite these promising results, further studies are necessary to understand the toxicity of these extracts and their constituentsfrom primary and secondary metabolism upon Ae. aegypti.Este trabalho objetivou avaliar a toxicidade dos extratos aquosos de sementes de 15 espécies de leguminosas contra larvas de Aedes aegypti. Foi realizada uma caracterização química e bioquímica parcial dos extratos aquosos e a avaliação da toxicidade aguda em camundongos. Os extratos de Amburana cearensis, Anadenanthera macrocarpa, Dioclea megacarpa, Enterolobium contortisiliquum e Piptadenia moniliformis causaram 100% de mortalidade depois de 1 a 3 h de exposição e mostraram valores de CL50 e CL90 entre 0,43 = 0,01 e 9,06 ± 0,12 e entre 0,71 = 0,02 e 13,03 = 0,15 mg/mL, respectivamente. Dentre os constituintes do metabolismo secundário, os extratos das sementes apresentaram taninos, fenóis, flavonas, flavonóis, xantonas, saponinas e alcalóides. Os extratos apresentaram alto teor de proteínas solúveis (0,98 to 7,71 mg/mL), lectina (32 to 256 UH/mL) e inibidor de tripsina (3,64 ± 0,43 to 26,19 = 0,05 gIT/kg de farinha). O perfil eletroforéticomostrou uma grande diversidade de proteínas, muitas dasquais já descritas como inseticidas. Os extratos mostraram baixa toxicidade ao camundongo (DL50 > 0,15 = 0,01 g/kg peso corporal), porém apesar desses resultados promissores, estudos posteriores são necessários para compreender a toxicidade desses extratos e de seus constituintes do metabolismo primário e secundário sobre Ae. aegypti.

Aedes aegypti is the major vector of 1 of the most concerning arboviruses of the world, the dengue fever. The only effective way of reducing the incidence of dengue fever is to control the vector mosquito, mainly by application of insecticides to its breeding places. This study was aimed at assessing the insecticidal activity of sodium anacardate, isolated from Brazilian cashew nut shell liquid (CNSL), against the eggs, 3rd instars or pupae of Ae. aegypti. In addition, the acute toxicity of sodium anacardate to mice was also investigated. Sodium anacardate showed toxicity against Ae. aegypti eggs (median effective concentration [EC50] = 162.93 +/- 29.93 microg/ml), larvae (median lethal concentration [LC50] = 55.47 +/- 3.0 microg/ml) and pupae (LC50 = 369.78 - 52.30 microg/ml). On the other hand, even at high dose (0.3 g/kg body weight), this compound did not cause any adverse effects on mice, suggesting that this compound is safe to mammals. Therefore, sodium anacardate may be a viable low-cost alternative to help combat Ae. aegypti.

This study assesses the nutritive and non-nutritive attributes of washed-up seaweeds from the Brazilian coast. It covers a broad diversity of species (24 red, nine green and four brown) with reasonable levels of proteins (10–14.8%), high ash contents (13–25%), low lipids (below 1%) and high carbohydrate contents (60%). Toxic and/or antinutritional factors were detected, such as low levels of lectins (32 and 64 HU/g of meal for chicken and rabbit trypsin-treated erythrocytes, respectively), tannins (59 mg/100 g), phytic acid (0.45%), high levels of trypsin inhibitors (99.0% inhibition) and -amylase inhibitors (70.5%). The 0/80% fraction showed moderate toxicity to mice (LD 50 of 63.8 mg kg −1 ). The presence of heavy metals such as cadmium (0.29 mg/100 g), chromium (0.23 mg/100 g), nickel (0.26 mg/100 g) and vanadium (3.56 mg/100 g) was also detected. Despite moderate toxicity and antinutritional limitations, washed-up seaweeds represent a potential food alternative for humans after appropriate processing and environmental remediation to guarantee food safety.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:49:34Z No. of bitstreams: 1 S0001-37652009000200007.pdf: 536011 bytes, checksum: 846642c2588a4bd24f90ee703c7ddc86 (MD5) Made available in DSpace on 2013-08-22T18:49:35Z (GMT). No. of bitstreams: 1 S0001-37652009000200007.pdf: 536011 bytes, checksum: 846642c2588a4bd24f90ee703c7ddc86 (MD5) Previous issue date: 2009-06-01 Made available in DSpace on 2013-09-30T19:39:57Z (GMT). No. of bitstreams: 2 S0001-37652009000200007.pdf: 536011 bytes, checksum: 846642c2588a4bd24f90ee703c7ddc86 (MD5) S0001-37652009000200007.pdf.txt: 42936 bytes, checksum: 25b8581e85c28f3681a9fb14d716430b (MD5) Previous issue date: 2009-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:14:42Z No. of bitstreams: 2 S0001-37652009000200007.pdf: 536011 bytes, checksum: 846642c2588a4bd24f90ee703c7ddc86 (MD5) S0001-37652009000200007.pdf.txt: 42936 bytes, checksum: 25b8581e85c28f3681a9fb14d716430b (MD5) Made available in DSpace on 2014-05-20T13:14:42Z (GMT). No. of bitstreams: 2 S0001-37652009000200007.pdf: 536011 bytes, checksum: 846642c2588a4bd24f90ee703c7ddc86 (MD5) S0001-37652009000200007.pdf.txt: 42936 bytes, checksum: 25b8581e85c28f3681a9fb14d716430b (MD5) Previous issue date: 2009-06-01 Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Neste trabalho, o extrato aquoso das sementes de Moringaoleifera (EASMO) foi avaliado quanto aos seus efeitos biológicos sobre ovos e larvas de Aedes aegypti no 3ºestágio de desenvolvimento e sua toxicidade sobre animais de laboratório(Daphnia magna, camundongos e ratos). O EASMO bruto revelou uma CL50 de 1.260 µg/mL, causando 99, 2 ± 2, 9% de mortalidade em 24 h na concentração de 5.200 µg/mL, embora o mesmo não tenha sido capaz de impedir a eclosão dos ovos. A atividade larvicida extinguiu-se após aquecimento do extrato a 80ºC/10 min. Diálises sucessivas do EASMO bruto resultaram em duas frações solúveis em água (Fração dializável, FD; Fração nãodializável, FND), dentre as quais apenas a FD mostrou ação larvicida. Testes de toxicidade aguda realizadosem dáfnias (CE50 de 188, 7 µg/mL) e camundongos (DL50 de446,5 mg/kg de peso corpóreo) evidenciaram baixa toxicidade. Apesar da hipertrofia tímica, o EASMO mostrou ser atóxicoapós tratamento subagudo via oral em ratos. Conclui-se, portanto, que o EASMO apresenta substâncias com capacida de larvicida contra Ae. aegypti, as quais possuem massa molecular aparente menor que 12 kDa e potencial tóxico moderado. In this work, biological effects of the water extract of Moringa oleifera seeds (WEMOS) were assessed on eggs and 3rd instar larvae of Aedes aegypti and on its toxicity upon laboratory animals (Daphnia magna, mice and rats). Crude WEMOS showed a LC50 value of 1260µg/mL, causing 99.2 ± 2.9% larvae mortality within 24 h at 5200µg/mL, though this larvicidal activity has been lost completely at 80ºC/10 min. WEMOS did not demonstrate capacity to prevent egg hatching. After extensive dialyses of the crude WEMOS into watersoluble dialyzable (DF) and nondyalizable (NDF) fractions, only DF maintained its efficacy to kill larvae. Acute toxicity evaluations on daphnids (EC50 of 188.7µg/mL) and mice (LD50 of 446.5 mg/kg body weight) pointed out to low toxicity. Despite the thymus hypertrophy, WEMOS revealed to be harmless in orally and subacutelytreated rats. In conclusion, WEMOS has thermostable bioactive compounds against Ae. aegypti larvae with apparent molecular mass lower than 12 kDa and moderately toxic potential. Universidade Federal do Ceará (UFC) Departamento de Biologia Universidade Federal do Ceará (UFC) Departamento de Análises Clínicas e Toxicológicas Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Defesa Fitossanitária Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Defesa Fitossanitária

This study assessed whether chemical analyses are sufficient to guarantee the safety of heat processing of soybeans (SB) for human/animal consumption. The effects of extrusion and dry-toasting were analyzed upon seed composition and performance of broiler chicks. None of these induced appreciable changes in protein content and amino acid composition. Conversely, toasting reduced all antinutritional proteins by over 85%. Despite that, the animals fed on toasted SB demonstrated a low performance (feed efficiency 57.8 g/100 g). Extrusion gave place to higher contents of antinutrients, particularly of trypsin inhibitors (27.53 g/kg flour), but animal performance was significantly (p < 0.05) better (feed efficiency 63.2 g/100 g). Upon the basis of chemical analyses, dry-toasting represents the treatment of choice. However, considering the results of the feeding trials, extrusion appears to be the safest method. In conclusion, in order to evaluate the reliability of any processing method intended to improve nutritional value, the combination of chemical and animal studies is necessary.

This work aims to review the nutritional properties of the Moringa oleifera tree, emphasizing its main constituents and nutritional applications for humans and animals. Moringa oleifera (Moringaceae) is a cosmopolitan tree that grows in many tropical countries showing uncountable folk uses due to its various nutritional and pharmacological applications. The young leaves, flowers and pods are common vegetables in the Asian diet. All parts of this plant are renewable sources of tocopherols (γ and α), phenolic compounds, β-carotene, vitamin C and total proteins, including the essential sulfur amino acids, methionine and cysteine. The seed protein and fat contents are higher than those reported for important grain legumes and soybean varieties, respectively. Unsaturated fatty acids, especially oleic acid, carbohydrates and minerals are present in the seed in reasonable amounts. In general, there are low concentrations of antinutritional factors in the plant, although the seeds possess glucosinolates (65.5µmol/g dry matter), phytates (41g/kg) and hemagglutination activity while the leaves have appreciable amounts of saponins (80g/kg), besides low quantity of phytates (21g/kg) and tannins (12g/kg). Taking into consideration the excellent nutritional properties, the low toxicity of the seeds and the excellent ability of the plant to adapt to poor soils and dry climates, Moringa oleifera can be an alternative to some leguminous seeds as a source of high-quality protein, oil and antioxidant compounds and a way to treat water in rural areas where appropriate water resources are not available.

This work aims to review the nutritional properties of the Moringa oleifera tree, emphasizing its main constituents and nutritional applications for humans and animals. Moringa oleifera (Moringaceae) is a cosmopolitan tree that grows in many tropical countries showing uncountable folk uses due to its various nutritional and pharmacological applications. The young leaves, flowers and pods are common vegetables in the Asian diet. All parts of this plant are renewable sources of tocopherols (γ and α), phenolic compounds, β-carotene, vitamin C and total proteins, including the essential sulfur amino acids, methionine and cysteine. The seed protein and fat contents are higher than those reported for important grain legumes and soybean varieties, respectively. Unsaturated fatty acids, especially oleic acid, carbohydrates and minerals are present in the seed in reasonable amounts. In general, there are low concentrations of antinutritional factors in the plant, although the seeds possess glucosinolates (65.5µmol/g dry matter), phytates (41g/kg) and hemagglutination activity while the leaves have appreciable amounts of saponins (80g/kg), besides low quantity of phytates (21g/kg) and tannins (12g/kg). Taking into consideration the excellent nutritional properties, the low toxicity of the seeds and the excellent ability of the plant to adapt to poor soils and dry climates, Moringa oleifera can be an alternative to some leguminous seeds as a source of high-quality protein, oil and antioxidant compounds and a way to treat water in rural areas where appropriate water resources are not available.O objetivo deste trabalho é fazer uma revisão sobre as propriedades nutricionais da planta Moringa oleifera, enfatizando seus principais constituintes e suas aplicações nutricionais para o homem e os animais. Moringa oleifera é uma planta que cresce em muitos países tropicais, possuindo inúmeros usos populares devido às suas aplicações nutricionais e farmacológicas. Na Ásia, suas folhas, flores e vagens são geralmente consumidas como vegetais. Todas as suas partes são fontes renováveis de compostos fenólicos, tocoferóis (γ e α), β-caroteno, vitamina C e proteínas totais, inclusive os aminoácidos essenciais sulfurados metionina e cisteína. Os conteúdos de proteínas e óleo nas sementes de Moringa oleifera são mais elevados que aqueles encontrados em legumes e em algumas variedades de soja, respectivamente. Ácidos graxos insaturados, principalmente o ácido oléico, carboidratos e minerais estão presentes nas sementes em quantidades razoáveis. No geral, a planta possui baixas concentrações de fatores antinutricionais, embora as sementes possuam glucosinolatos (65,5µmol/g), fitatos (41g/kg) e atividade hemaglutinante, enquanto as folhas têm apreciáveis quantidades de saponinas (80g/kg), além de fitatos (21g/kg) e taninos (12g/kg). Levando em consideração as excelentes propriedades nutricionais, a baixa toxicidade das sementes e a excelente habilidade da planta de se adaptar a solos pobres e a climas áridos, a Moringa oleifera pode ser uma alternativa ao consumo de sementes leguminosas, como fonte de proteínas de alta qualidade, de óleo e de compostos antioxidantes. Pode ser usada, ainda, como uma maneira de tratar água em áreas rurais onde recursos hídricos adequados não estão disponíveis.

Mo-CBP3 is an antifungal protein produced by Moringa oleifera which has been investigated as potential candidate for developing transgenic crops. Before the use of novel proteins, food safety tests must be conducted. This work represents an early food safety assessment of Mo-CBP3, using the two-tiered approach proposed by ILSI. The history of safe use, mode of action and results for amino acid sequence homology using the full-length and short contiguous amino acids sequences indicate low risk associated to this protein. Mo-CBP3 isoforms presented a reasonable number of alignments (>35% identity) with allergens in a window of 80 amino acids. This protein was resistant to pepsin degradation up to 2 h, but it was susceptible to digestion using pancreatin. Many positive attributes were presented for Mo-CBP3. However, this protein showed high sequence homology with allergens and resistance to pepsin digestion that indicates that further hypothesis-based testing on its potential allergenicity must be done. Additionally, animal toxicity evaluations (e.g. acute and repeated dose oral exposure assays) must be performed to meet the mandatory requirements of several regulatory agencies. Finally, the approach adopted here exemplified the importance of performing an early risk assessment of candidate proteins for use in plant transformation programs.

Insecticidal Cry proteins from Bacillus thuringiensis (Bt) have been exploited in the development of genetically modified (GM) crops for pest control. However, several pests are still difficult to control such as the coleopteran boll weevil Anthonomus grandis. By applying in vitro molecular evolution to the cry8Ka1 gene sequence, variants were generated with improved activity against A. grandis. Among them, Cry8Ka5 mutant protein showed coleoptericidal activity 3-fold higher (LC50 2.83 μg/mL) than that of the original protein (Cry8Ka1). Cry8Ka5 has been used in breeding programs in order to obtain coleopteran-resistant cotton plants. Nevertheless, there is some concern in relation to the food safety of transgenic crops, especially to the heterologously expressed proteins. In this context, our research group has performed risk assessment studies on Cry8Ka5, using the tests recommended by Codex as well as tests that we proposed as alternative and/or complementary approaches. Our results on the risk analysis of Cry8Ka5 taken together with those of other Cry proteins, point out that there is a high degree of certainty on their food safety. It is reasonable to emphasize that most safety studies on Cry proteins have essentially used the Codex approach. However, other methodologies would potentially provide additional information such as studies on the effects of Cry proteins and derived peptides on the indigenous gastrointestinal microbiota and on intestinal epithelial cells of humans. Additionally, emerging technologies such as toxicogenomics potentially will offer sensitive alternatives for some current approaches or methods.