Toxicity and Anti-Inflammatory Effects of Agave sisalana Extract Derived from Agroindustrial Residue.

BACKGROUND: In several countries, the leaf juice of Agave sisalana (also known as sisal) is widely used topically, especially as an antiseptic, and orally for the treatment of different pathologies. However, in Brazil, which is the largest producer of Agave sisalana, its residue, which represents the majority of its weight, has been thrown away. For this reason, the determination of the pharmacological and toxicological potentials of sisal residue and its possible therapeutic use is seen as a way to contribute to the sustainable development and social promotion of the largest producer of sisal in Brazil, the interior of Bahia State, which is among the poorest areas in the country. Given the scarcity of available scientific studies on the pharmacological and toxicological properties of sisal residue juice, this study aimed to promote the acid hydrolysis of this juice to potentiate the anti-inflammatory effect already described in the literature. Furthermore, it aimed to evaluate the toxicological profile of the hydrolyzed extract (EAH) and to determine its acute toxicity, as well as its side effects on the reproductive aspects of rats. METHOD: The anti-inflammatory effect of EAH was evaluated in vitro using the induction of hemolysis by hypotonic solution and in vivo in rats using the carrageenan-induced paw edema test and the xylene-induced ear edema test. The acute toxicity, resulting from a single-dose administration, was investigated for some manifestation of toxic symptoms related to motor control and consciousness in rats. At a concentration of 100 mg/kg, by repeated doses, the reproductive toxicity effects of EAH in rats were assessed. RESULTS: In vitro anti-inflammatory activity was positive using the human red blood cell membrane stabilization method. In both in vivo tests used to assess the anti-inflammatory activity, EAH (at three doses) significantly inhibited edema when compared to the control group. At a dose of 50 mg/kg, EAH exhibited a greater effect than indomethacin, a nonsteroidal anti-inflammatory drug with known activity. In vivo toxicological studies have shown that EAH does not present toxic effects when administered orally in a single dose, up to 1000 mg/kg. Finally, EAH promoted a gonadotoxic effect and increased the embryonic mortality rate after implantation. CONCLUSIONS: It is suggested that the anti-edematogenic effect of the acid hydrolysis extract from sisal juice is due to the high concentration of steroidal sapogenins. Therefore, this extract can be considered a potential new anti-inflammatory or even an important sapogenin source for the development of steroidal glucocorticoids. However, further studies are needed to elucidate the chemical composition of sisal juice. Regarding toxicology studies, EAH did not show cytotoxic and clastogenic potentials, but it presented a powerful reproductive toxic effect in rats.

Medical applications of clustered regularly interspaced short palindromic repeats (CRISPR/Cas) tool: a comprehensive overview

Since the discovery of the double helix and the introduction of genetic engineering, the possibility to develop new strategies to manipulate the genome has fascinated scientists around the world. Currently scientists have the knowledge and ability to edit the genomes. Several methodologies of gene editing have been established, all of them working like "scissor", creating double strand breaks at specific spots. The introduction of a new technology, which was adapted from the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas bacterial immune system, has revolutionized the genetic therapy field, as it allows a much more precise editing of gene than the previously described tools and, therefore, to prevent and treat disease in humans. This review aims to revisit the genome editing history that led to the rediscovery of the CRISPR/Cas technology and to explore the technical aspects, applications and perspectives of this fascinating, powerful, precise, simpler and cheaper technology in different fields

Medical applications of clustered regularly interspaced short palindromic repeats (CRISPR/Cas) tool: a comprehensive overview

Since the discovery of the double helix and the introduction of genetic engineering, the possibility to develop new strategies to manipulate the genome has fascinated scientists around the world. Currently scientists have the knowledge and ability to edit the genomes. Several methodologies of gene editing have been established, all of them working like "scissor", creating double strand breaks at specific spots. The introduction of a new technology, which was adapted from the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas bacterial immune system, has revolutionized the genetic therapy field, as it allows a much more precise editing of gene than the previously described tools and, therefore, to prevent and treat disease in humans. This review aims to revisit the genome editing history that led to the rediscovery of the CRISPR/Cas technology and to explore the technical aspects, applications and perspectives of this fascinating, powerful, precise, simpler and cheaper technology in different fields

Toxicological, genotoxic and antioxidant potential of pyrostegia venusta / Potencial toxicológico, genotóxico e antioxidante de Pyrostegia venusta

Pyrostegia venusta is usually found in the secondary growth of the Atlantic forests, and in the Brazilian Savanna. Flowers and leaves of this plant are used in folk remedies for treating a wide variety of healthy conditions, this way is important evaluate its safety and antioxidant potential for this applications. For this, was made a ethanolic extract from its flowers and analyzed with toxicological,genotoxicity and antioxidant tests, the toxicological analysis was made by reproductive toxicity in rats and clatogenicity/aneugenicity in human lymphocytes. The genotoxicity was studied by micronucleus test mice bone marrow. The antimutagenic test in root cells of Allium cepa, the antioxidant assays used was DPPH, FRAP, Lipid Perxidation and REM, beyond of that the extract was analyzed in HPLC showing the profile of its compounds. The toxicological analysis showed that P. venusta has no negative significant effect on reproductive and cellular level. The micronucleus test in mouse bone marrow, the extract protected cells from cyclophosphamide, mutagenic compound, in a similar way. The A. cepa test showed that the extract reduced chromosomal disorders formations. The antioxidant activity of extract was significant, except in REM test. The phytochemical analysis showed the presence of flavonoids compounds. P. venusta extract does not present reproductive toxicity and genotoxic effects. However, the extract of this species showed antigenotoxic and antioxidant potential, possibly due to the different flavonoid compounds present in its extract.

Pyrostegia venusta é geralmente encontrada no crescimento secundário das florestas atlânticas e na savana brasileira. Flores e folhas desta planta são utilizadas em remédios populares para tratar uma grande variedade de doenças, desta forma é importante avaliar a segurança e o potencial antioxidante para estas aplicações. Para tanto, o extrato etanólico das flores foi avaliado com testes toxicológicos, genotóxicos e antioxidants. A análise toxicológica foi realizada por meio da toxicidade reprodutiva em ratos e a clatogenicidade/aneugenicidade em linfócitos humanos, a genotoxicidade foi estudada por teste de micronúcleo em medula óssea de camundongo. A antimutagenicidade em células da raiz de Allium cepa. Os ensaios antioxidantes utilizados foram DPPH, FRAP, TARBS e MRE. O extrato foi analisado em HPLC. A análise toxicológica reprodutiva mostrou que P. venusta não tem efeito negativo sobre o nível reprodutivo e cellular. No teste do micronúcleo o extrato protegeu as células da ciclofosfamida, um composto mutagênico. O teste de A. cepa mostrou que o extrato reduziu as formações dos distúrbios cromossômicos. A atividade antioxidante do extrato foi significativa, exceto no teste REM. A análise fitoquímica mostrou a presença de compostos flavonoídicos. O extrato de P. venusta não apresenta toxicidade reprodutiva e efeitos genotóxicos. No entanto, o extrato desta espécie apresentou potencial antigenotóxico e antioxidante, possivelmente devido aos diferentes compostos flavonoídicos presentes em seu extrato.

The human papillomavirus (HPV)-related cancer biology: an overview

Despite the novel diagnostic methods and therapies implemented in oncology, the number of patients that succumb by the cancer remains high globally. Currently studies point out that 20–25% of all human malignancies are related to micro-organism infections. Among these cancer-related pathogens, the human papillomavirus (HPV) has a prominent position, since the virus is responsible for about 30% of all infectious agent-related cancers. Thus, an amount of cancers could be avoided by means prophylactic and/or therapeutic measures. However, these measures required a holistic comprehension about HPV-related cancer biology. Based on this, this review aims to summarize the last evidences of HPV on cancer biology (from initiation to metastasis), focus on molecular and biochemical deregulations associated with viral infection, and discuss the viral etiology in different malignancies.

Analysis of antioxidant, cytotoxic and mutagenic potential of Agave sisalana Perrine extracts using Vero cells, human lymphocytes and mice polychromatic erythrocytes

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins.

The human papillomavirus (HPV)-related cancer biology: an overview

Despite the novel diagnostic methods and therapies implemented in oncology, the number of patients that succumb by the cancer remains high globally. Currently studies point out that 20–25% of all human malignancies are related to micro-organism infections. Among these cancer-related pathogens, the human papillomavirus (HPV) has a prominent position, since the virus is responsible for about 30% of all infectious agent-related cancers. Thus, an amount of cancers could be avoided by means prophylactic and/or therapeutic measures. However, these measures required a holistic comprehension about HPV-related cancer biology. Based on this, this review aims to summarize the last evidences of HPV on cancer biology (from initiation to metastasis), focus on molecular and biochemical deregulations associated with viral infection, and discuss the viral etiology in different malignancies.

Analysis of antioxidant, cytotoxic and mutagenic potential of Agave sisalana Perrine extracts using Vero cells, human lymphocytes and mice polychromatic erythrocytes

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins.

Análise do potencial mutagênico dos esteroides anabólicos androgênicos (EAA) e da l-carnitina mediante o teste do micronúcleo em eritrócitos policromáticos / Analysis of mutagenic potential of the anabolic androgenic steroids (AAS) and l-carnitine by the micronucleus test in polychromatic erythrocytes / Análisis del potencial mutagénico de los esteroides anabólicos androgénicos (EAA) y de la l-carnitina mediante la prueba del micronúcleo en eritrocitos policromáticos

INTRODUÇÃO: Os esteroides anabólicos androgênicos são usados por pessoas que desejam aumentar sua massa muscular para obter um melhor desempenho nos esportes ou melhorar a aparência física. Os EAA são derivados sintéticos da testosterona, capazes de promover a hipertrofia das fibras musculares, aumentando a síntese proteica intracelular. A L-carnitina é um suplemento alimentar empregado para aumentar a produção energética por meio da oxidação de ácidos graxos. Embora haja trabalhos mostrando as propriedades fisiológicas dessas drogas, há poucos estudos sobre o potencial mutagênico das mesmas. OBJETIVOS: Este trabalho avaliou a clastogenicidade e genotoxicidade do decanoato de nandrolona, decanoato de testosterona e da L-carnitina, em diferentes tratamentos, através do teste do micronúcleo em eritrócitos policromáticos de ratos Wistar. MÉTODOS: Os animais foram submetidos a diferentes concentrações e associações de EAA. O controle positivo recebeu ciclofosfamida 50 mg/kg através de injeção intraperitoneal e o controle negativo, 1 ml de soro fisiológico por gavagem. Os ratos foram sacrificados após 36 horas da última aplicação, tendo seus fêmures removidos e a medula óssea extraída. O material foi homogeneizado e centrifugado. O botão de células foi pipetado e transferido para as lâminas, que foram coradas com Giemsa. Foram contados 1.000 eritrócitos policromáticos por animal, observando a frequência de micronúcleos. RESULTADOS: Foi realizado o teste de Kruskal-Wallis, com nível de significância de 5%, que demostrou que o decanoato de nandrolona - três doses de 0,2 mg/kg e 0,6 mg/kg, oito doses de 7,5 mg/kg, L-carnitina - sete doses de 0,4 ml/250g e 1,5 ml/250g, decanoato de testosterona - 28 doses de 0,075 mg/kg, decanoato de nandrolona - oito doses de 7,5 mg/kg associado a L-carnitina 1 ml e decanoato de nandrolona - oito doses de 7,5 mg/kg associado à decanoato de testosterona - oito doses de 7,5 mg/kg apresentaram potencial mutagênico. CONCLUSÃO: Os tratamentos revelaram-se clastogênicos, não sendo indicado como recurso ergogênico.

INTRODUCTION: Anabolic androgenic steroids (AAS) are frequently used by people whose aim to increase muscle mass to obtain a better performance in sports or improve physical appearance. AAS are synthetic derivatives of testosterone, able to promote muscle fibers hypertrophy, increasing intracellular protein synthesis. L-carnitine is a food supplement used to increase energetic production by means of fat acids oxidation. Although there are several works about physiological properties of these drugs, there are few studies about their mutagenic potential. OBJECTIVES: This work evaluated the clastogenicity and genotoxicity of nandrolone decanoate, testosterone decanoate and L-carnitine, in different treatments through the micronucleus test in polychromatic erythrocytes of Wistar rats. METHODS: The animals were submitted to different concentrations and associations of AAS. The positive control received cyclophosphamide 50 mg/kg by intraperitoneal injection and negative control, one ml of saline solution by gavage. The rats were sacrificed after 36 hours of latest application, having the femurs removed and the bone marrow extracted. Material was homogenized and centrifuged. Button cell was pipetted and transferred to slides, which were stained by Giemsa. 1,000 polychromatic erythrocytes were counted per animal, noting the frequency of micronuclei. RESULTS: The Kruskal-Wallis test was performed, with a significance level of 5%, which demonstrated that nandrolone decanoate - three doses of 0,2 mg/kg and 0,6 mg/kg, eight doses of 7,5 mg/kg, L-carnitine - seven doses of 0,4 ml/250 g and 1,5 ml/250 g, testosterone decanoate - 28 doses of 0,075 mg/kg, nandrolone decanoate - eight doses of 7,5 mg/kg associated to L-carnitine and 1 mL and nandrolone decanoate - eight doses of 7,5 mg/kg associated to testosterone decanoate - eight doses of 7,5 mg/kg, showed mutagenic potential. CONCLUSION: The treatments proved to be clastogenic, not being indicated like ergogenic aid.

INTRODUCCIÓN: Los esteroides anabólicos androgénicos son usados por personas que quieren aumentar su masa muscular a fin de obtener un mejor desempeño en los deportes o para mejorar la apariencia física. Los EAA son derivados sintéticos de la testosterona, que pueden causar la hipertrofia de las fibras musculares, aumentando la síntesis proteica intracelular. La L-carnitina es un suplemento alimenticio utilizado para hacer crecer la producción energética por medio de la oxidación de ácidos grasos. Aunque hay trabajos que muestran las propiedades fisiológicas de esos medicamentos, hay pocos estudios sobre el potencial mutagénico de estos. OBJETIVOS: Este trabajo evaluó la clastogenicidad y la genotoxicidad del decanoato de nandrolona, decanoato de testosterona y de la L-carnitina, en diversos tratamientos, mediante la prueba del micronúcleo en eritrocitos policromáticos de ratones Wistar. MÉTODOS: Los animales fueron sometidos a diferentes concentraciones y asociaciones de EAA. El control positivo recibió ciclofosfamida 50 mg/kg a través de inyección intraperitoneal, y el control negativo 1 ml de suero fisiológico por gavaje. Los ratones fueron sacrificados después de 36 horas de la última aplicación, teniendo sus fémures removidos y la médula ósea extraída. El material fue homogeneizado y centrifugado. El botón de células fue tomado con pipetas y transferido para las láminas, a las cuales se les dio color con Giemsa. Fueron contados 1.000 eritrocitos policromáticos por animal, observándose la frecuencia de micronúcleos. RESULTADOS: Se realizó la prueba de Kruskal-Wallis, con nivel de significancia de 5%, la cual demostró que el decanoato de nandrolona - tres dosis de 0,2 mg/kg y 0,6 mg/kg, ocho dosis de 7,5 mg/kg, L-carnitina - siete dosis de 0,4 ml/250g y 1,5 ml/250g, decanoato de testosterona - 28 dosis de 0,075 mg/kg, decanoato de nandrolona - ocho dosis de 7,5 mg/kg asociado a L-carnitina 1 ml y decanoato de nandrolona - ocho dosis de 7,5 mg/kg vinculado al decanoato de testosterona - ocho dosis de 7,5 mg/kg, presentaron potencial mutagénico. CONCLUSIÓN: Los tratamientos se revelaron como siendo clastogénicos, no indicándose como recurso ergogénico.

Genetic variability of Herpailurus yagouaroundi, Puma concolor and Panthera onca (Mammalia, Felidae) studied using Felis catus microsatellites

We used four microsatellite loci (Fca08, Fca45, Fca77 and Fca96) from the domestic cat, Felis catus, to investigate genetic variability in specimens of Herpailurus yagouaroundi (jaguarundi, otter cat, eyra), Puma concolor (cougar, mountain lion, puma) and Panthera onca (jaguar) held in various Brazilian zoos. Samples of DNA from the cats were PCR amplified and then sequenced before being analyzed using the CERVUS program. Our results show a mean polymorphic information content (PIC) of 0.83 for H. yagouaroundi, 0.66 for P. concolor and 0.69 for P. onca and a mean of 10.3 alleles for the Fca08 locus, 5.3 for Fca 45, 9 for Fca 77 and 14 for Fca 96. These results indicate a relatively high level of genetic diversity for the specimens studied.