Própolis vermelha e ação antifúngica: potencialidades e desafios / Red própolis and antifungal activity: potentialities and challenges / Propóleo rojo y acción antifúngica: potencialidades y desafíos

Introdução: a própolis é uma composição resinosa produzida por abelhas e utilizada em suas colmeias contra microrganismos. Existem diversos tipos desse composto, sendo o de coloração vermelha o último espécime relatado na literatura. Assim, dentre suas aplicabilidades, a atividade antifúngica da própolis vermelha tem sido explorada com vistas a ampliar sua ação terapêutica. Objetivo: explorar estudos acerca da ação antifúngica da própolis vermelha, identificando suas potencialidades e desafios. Metodologia: foi realizada uma revisão integrativa nas bases de dados bibliográficos MEDLINE (via PubMed), SciELO e Google Acadêmico, complementada por uma diligência nas bases de ensaios clínicos ReBEC e Clinical Trials. Em seguida todos os estudos selecionados foram explorados para obtenção do cenário atual sobre o tema. Resultados: foram incluídos 08 estudos, sendo 01 deles um ensaio clínico. Os estudos comprovam a ação antifúngica da própolis vermelha, principalmente contra Candida spp. e Paracoccidioides brasiliensis, e evidenciam a maior potência fungicida deste composto em detrimento de outros tipos de própolis. Conclusão: a ação antifúngica da própolis vermelha mostra-se uma potencialidade em diversos estudos. Entretanto, o volume de pesquisas científicas relativas a esse tema é insuficiente e a complexidade desse composto configura-se como um desafio à sua aplicabilidade.

Introduction: propolis is a resinous composition produced by compounds and used in their hives against microorganisms. There are several types of this compound, the red one is the last specimen reported in the literature. Thus, among its applicability, the antifungal activity of red propolis has been explored as a path to expand its therapeutic action. Objective: to explore studies about the antifungal action of red propolis, identifying its potentialities and challenges. Methodology: Na integrative review was carried out in the bibliographic databases MEDLINE (via PubMed), SciELO and Google Scholar, complemented by a diligence in ReBEC and Clinical Trials databases. Then, all selected studies were explorers to obtain the current scenario on the subject. Results: 08 studies were included, which 01 of them was a clinical trial. Studies prove the antifungal action of red propolis, mainly against Candida spp. and Paracoccidioides brasiliensis, and show the greater fungicidal power of this compound compared to other types of propolis. Conclusion: the antifungal action of red propolis shows potential in several studies. However, the volume of scientific research on this theme is insufficient and the complexity of this compound represents a challenge to its applicability.

Introducción: el propóleo es una composición resinosa producida por las abejas y utilizada en sus colmenas contra los microorganismos. Existen varios tipos de este compuesto, siendo el rojo el último ejemplar reportado en la literatura. Así, entre sus posibilidades de aplicación, se ha explorado la actividad antifúngica del propóleo rojo con vistas a ampliar su acción terapéutica. Objetivo: explorar estudios sobre la acción antifúngica del propóleo rojo, identificando sus potencialidades y desafíos. Metodología: Se realizó una revisión en las bases de datos bibliográficas MEDLINE (vía PubMed), SciELO y Google Scholar, complementada con una diligencia en las bases de datos de ensayos clínicos ReBEC y Clinical Trials. Luego se exploraron todos los estudios seleccionados para obtener el escenario actual sobre el tema. Resultados: Se incluyeron 08 estudios, 01 de los cuales fue un ensayo clínico. Los estudios demuestran la acción antifúngica del propóleo rojo, principalmente contra Candida spp. y Paracoccidioides brasiliensis, y muestran el mayor poder fungicida de este compuesto en detrimento de otros tipos de propóleos. Conclusión: la acción antifúngica del propóleo rojo muestra potencial en varios estudios. Sin embargo, el volumen de investigación científica sobre este tema es insuficiente y la complejidad de este compuesto representa un desafío para su aplicabilidad.

Biomimetics and the restoration of the endodontically treated tooth / Biomimética e a restauração do dente tratado endodonticamente

Although much progress has been obtained in terms of the Endodontic treatment, the literature shows that true success can be only achieved with adequate coronal seal to avoid bacterial contamination, and protect the tooth structure from fracture. There are many options available to the clinician to restore the endodontically treated tooth; however, there is not much evidence available on what alternative is better than another. This review will critically present the current knowledge on restorative choices, including posts and endocrowns, showing advantages and disadvantages of different treatment forms. With this knowledge, we will also introduce the concept of biomimetics to endodontically treated teeth, and how the nature of their remaining tooth structure can benefit from this approach. This concept entails the use of mechanisms and biologically produced materials to restore a tooth in a way that would mimic its natural structure, with the purpose of achieving better long-term prognosis (AU)

Embora tenha se obtido progresso em relação ao tratamento endodôntico, a literatura mostra que o sucesso real só pode ser atingido com o selamento coronal adequado, para evitar-se a contaminação bacteriana e proteger-se a estrutura dental de fraturas. Há muitas opções disponíveis para o clínico para a restauração do dente tratado endodonticamente; entretanto, não há muita evidência disponível sobre qual alternativa é melhor que a outra. Esta revisão apresentará criticamente o conhecimento atual sobre opções restauradoras, incluindo retentores intraradiculares e endocrowns, mostrando vantagens e desvantagens das diferentes formas de tratamento. Com esse conhecimento, também introduziremos o conceito de biomimética, uma vez que dentes tratados endodonticamente, devido a natureza de sua estrutura dental remanescente, podem se beneficiar desta abordagem. Esse conceito envolve o uso de mecanismos e materiais produzidos biologicamente para restaurar um dente de forma a imitar a estrutura natural, com o objetivo de alcançar melhor prognóstico de longo-prazo.(AU)

Mutagenic Evaluation of Fluoxetine and Fluoxetine-Galactomannan Complex Through the Analysis of Chromosomal Aberrations in Human Peripheral Leukocytes and Salmonella typhimurium/Microssome Assay / Avaliação Mutagênica de Fluoxetina e Complexo Fluoxetina-Galactomanana Através da Análise de Aberrações Cromossômicas em Leucócitos Periféricos Humanos e em Salmonella typhimurium / Ensaio Microssômico

Objectives: The purpose of this study was to evaluate the mutagenic potential of fluoxetine and fluoxetine-galactomannan. Methods: Chromosomal aberration test and Salmonella typhimurium/microsome mutagenicity assay. Results: The results showed that fluoxetine (250 µg/mL) can cause chromosomal breaks of treated leukocytes and increase the frequency of reversion of the tester strains of S. typhimurium / microsome assay only at the highest concentration (5 mg/mL), while fluoxetine encapsulated in galactomannan did not cause these changes (leukocytes and S. typhimuriums strains). Conclusion: In summary, fluoxetine showed a mutagenic effect detectable only at high concentrations in both eukaryotic and prokaryotic models. Furthermore, the fluoxetine/galactomannan complex, in this first moment, prevented the mutagenicity attributed to fluoxetine, emphasizing that the present encapsulation process can be an alternative in preventing these effects in vitro.

Objetivos: avaliar o potencial mutagênico da fluoxetina e da fluoxetina-galactomanana. Métodos: Teste de aberração cromossômica e ensaio de mutagenicidade de Salmonella typhimurium /microssoma. Resultados: a fluoxetina (250 µg/mL) pode causar quebras cromossômicas de leucócitos tratados e aumentar a frequência de reversão das cepas testadoras de S. typhimurium /microssoma apenas na concentração mais alta (5 mg/mL), enquanto a fluoxetina encapsulada em galactomanano não causou essas alterações (leucócitos e cepas de S. typhimurium). Conclusão: a fluoxetina mostrou um efeito mutagênico detectável apenas em altas concentrações em modelos eucarióticos e procarióticos. Além disso, o complexo fluoxetina/galactomanan, neste primeiro momento, evitou a mutagenicidade atribuída à fluoxetina, ressaltando que o presente processo de encapsulamento pode ser uma alternativa na prevenção desses efeitos in vitro.

Evaluation of interactions of silibinin with the proteins ALS3 and SAP5 against Candida albicans / Avaliação das interações da silibinina com as proteínas ALS3 e SAP5 contra Candida albicans

Objective: to evaluate the molecular interaction of silibinin with the targets ALS3 and SAP5. Methodology: Molecular docking protocols were conducted to analyze the binding interaction of silibinin with ALS3 and SAP5. Results: Eleven interactions of ALS3 with silibinin and four with fluconazole were found, while six interactions were observed of SAP5 with silibinin and four with fluconazole. Conclusion: Molecular docking between silibinin and ALS3 identified important interactions, but no significant interactions were observed with SAP5, even though silibinin can exhibit affinity and interactions with other SAP5 sites.

Objetivo: Avaliar a interação molecular da silibinina com os alvos ALS3 e SAP5. Metodologia: Protocolos de docking molecular foram conduzidos para analisar a interação de ligação da silibinina com ALS3 e SAP5. Resultados: Foram encontradas onze interações de ALS3 com silibinina e quatro com fluconazol, enquanto seis interações foram observadas de SAP5 com silibinina e quatro com fluconazol. Conclusão: Docking molecular entre silibinina e ALS3 identificou interações importantes, mas não foram observadas interações significativas com SAP5, embora a silibinina possa apresentar afinidade e interações com outros sítios SAP5.

Study of the interactions of di- and tri-terpenes from Stillingia loranthacea with the enzyme NSP16-NSP10 of SARS-CoV-2 / Estudo das interações de di- e tri-terpenos de Stillingia loranthacea com a enzima NSP16-NSP10 de SARS-CoV-2

Objective: This study aimed to evaluate the interactions of di- and tri-terpenes from Stillingia loranthacea with the enzyme NSP16-NSP10 of SARS-CoV-2, important for viral replication. Methods: The molecular docking technique was used to evaluate this interaction. Results: The analysis showed that the evaluated compounds obtained RMSD values of 0.888 to 1.944 Å and free energy of -6.1 to -9.4 kcal/mol, with the observation of hydrogen bonds, salt bridges, and pi-sulfur, pi-alkyl, and hydrophobic interactions. Conclusion: Thus, the results obtained show the potential of the compounds analyzed against the selected target. Since computer simulations are only an initial step in projects for the development of antiviral drugs, this study provides important data for future research.

Objetivo: avaliar as interações de di- e tri-terpenos de Stillingia loranthacea com a enzima NSP16-NSP10 de SARS-CoV-2, importante para a replicação viral. Métodos: A técnica de docking molecular foi utilizada para avaliar essa interação. Resultados: A análise mostrou que os compostos avaliados obtiveram valores de RMSD de 0,888 a 1,944 Å e energia livre de -6,1 a -9,4 kcal/mol, observando-se ligações de hidrogênio, pontes salinas e pi-enxofre, pi-alquil, e interações hidrofóbicas. Conclusão: Assim, os resultados obtidos mostram o potencial dos compostos analisados frente ao alvo selecionado. Como as simulações computacionais são apenas um passo inicial nos projetos de desenvolvimento de medicamentos antivirais, este estudo fornece dados importantes para pesquisas futuras.

Virtual screening based on molecular docking of lysosomotropic compounds as therapeutic agents for COVID-19 / Triagem virtual baseada no encaixe molecular de compostos lisossomotrópicos como agentes terapêuticos para COVID-19

Objective: Analyze lysosomotropic agents and their action on COVID-19 targets using the molecular docking technique. Methods: Molecular docking analyses of these lysosomotropic agents were performed, namely of fluoxetine, imipramine, chloroquine, verapamil, tamoxifen, amitriptyline, and chlorpromazine against important targets for the pathogenesis of SARS-CoV-2. Results: The results revealed that the inhibitors bind to distinct regions of Mpro COVID-19, with variations in RMSD values from 1.325 to 1.962 Å and binding free energy of -5.2 to -4.3 kcal/mol. Furthermore, the analysis of the second target showed that all inhibitors bonded at the same site as the enzyme, and the interaction resulted in an RMSD variation of 0.735 to 1.562 Å and binding free energy ranging from -6.0 to -8.7 kcal/mol. Conclusion: Therefore, this study allows proposing the use of these lysosomotropic compounds. However, these computer simulations are just an initial step toward conceiving new projects for the development of antiviral molecules.

Objetivo: aAnalisar agentes lisossomotrópicos e sua ação em alvos de COVID-19 usando a técnica de docking molecular. Métodos: Foram realizadas análises de docagem molecular destes agentes lisossomotrópicos, nomeadamente de fluoxetina, imipramina, cloroquina, verapamil, tamoxifeno, amitriptilina e clorpromazina contra alvos importantes para a patogenia do SARS-CoV-2. Resultados: Os resultados revelaram que os inibidores se ligam a regiões distintas do Mpro COVID-19, com variações nos valores de RMSD de 1.325 a 1.962 Å e energia livre de ligação de -5,2 a -4,3 kcal/mol. Além disso, a análise do segundo alvo mostrou que todos os inibidores se ligaram no mesmo sítio da enzima, e a interação resultante em uma variação de RMSD de 0,735 a 1.562 Å e energia livre de ligação variando de -6,0 a -8,7 kcal/mol. Conclusão: Portanto, este estudo permite propor o uso desses compostos lisossomotrópicos. No entanto, essas simulações em computador são apenas um passo inicial para a concepção de novos projetos para o desenvolvimento de moléculas antivirais.

Gum arabic and red propolis protecteting colorectal preneoplastic lesions in a rat model of azoxymethane

Abstract Purpose: To evaluate red propolis, gum arabic and L-lysine activity on colorectal preneoplastic lesions induced by azoxymethane (AOM). Methods: The study featured 4 control groups (I-IV) and 4 experimental groups (V-VIII), totaling 48 rats. Once a week for 2 weeks, animals on control groups received saline, while animals in experimental groups received azoxymethane (15 mg/kg i.p.). The follow up along 16 weeks included daily oral gavage to administer water (I and V), L-lysine (150 mg/kg)(II and VI), própolis (100mg/5ml/kg)(III and VII), or gum arabic (5ml/kg)(IV and VIII). Was performed surgery on the animals in the end of this time in order to collect blood for biological assays (TBARS, GSH), followed by their sacrifice to tissue extract. Results: Oxidative stress (TBARS) and the number of aberrant crypt foci (ACF) in distal colon were lower using própolis (p<0.01 for both parameters). Gum arabic reduced preneoplastic lesions (ACF ≤ 4 crypts) on distal colon and on the entire colon (p<0.05). Conclusions: Red propolis reduced AOM-induced oxidative stress (TBARS) and total number of ACF in the distal colon. L-lysine neither protected against nor enhanced AOM-induced ACF. Gum arabic reduced the number of ACF.

Effects of mineral trioxide aggregate, BiodentineTM and calcium hydroxide on viability, proliferation, migration and differentiation of stem cells from human exfoliated deciduous teeth

Abstract Objective: The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. Material and Methods: SHED were cultured for 1 - 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 μm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. Results: MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Conclusion: Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population.

EPIDEMIOLOGIA DOS ACIDENTES OFÍDICOS NOTIFICADOS EM UM CENTRO DE ASSISTÊNCIA TOXICOLÓGICA DE 2011 A 2015 / EPIDEMIOLOGY OF OPHIDIAN ACCIDENTS NOTIFIED TO A TOXICOLOGIC ASSISTANCE CENTER FROM 2011 TO 2015 / PIDEMIOLOGÍA DE LOS ACCIDENTES OFÍDICOS NOTIFICADOS EN UN CENTRO DE ASISTENCIA TOXICOLÓGICA DE 2011 HASTA 2015

O Brasil é um dos países sul-americanos com maior prevalência de acidentes ofídicos por ano, entretanto esses acontecimentos são considerados negligenciados por países tropicais e subtropicais em desenvolvimento. O objetivo deste artigo foi traçar o perfil epidemiológico dos acidentes ofídicos notificados no período 2011-2015, em um Centro de Assistência Toxicológica. Para tanto, fez-se uma pesquisa transversal de abordagem quantitativa realizada com dados epidemiológicos registrados e documentados nas fichas de notificação e de atendimento de acidentes ofídicos. Foram coletadas, para análise, nove variáveis, e os dados encontrados foram analisados por meio de estatística descritiva. Assim, os resultados indicaram a notificação de 183 casos, sendo 19,13% relativos aos acidentes botrópicos. O sexo masculino foi o mais acometido, com 67,76%, e predominou a faixa etária de 10 a 49 anos (73,22%), tendo os membros inferiores como a região anatômica mais afetada (74,32%). A maioria das ocorrências foi na zona rural (54,10%), principalmente agricultores (26,23%), na ocasião em que exerciam sua ocupação (13,66%). Concluiu-se que é necessário o desenvolvimento de pesquisas que contribuam para o (re)conhecimento do perfil epidemiológico desses acidentes como forma de favorecer o desenvolvimento de estratégias preventivas, bem como reforçar a importância da identificação das espécies responsáveis pelos acidentes para que haja um tratamento efetivo e menor risco de ocorrência de óbitos e sequelas nos indivíduos acometidos.

Brazil is one of the South American countries with bigger prevalence of ophidian accidents by year, however these events are considered neglected by tropical and subtropical countries in development process. The objective of this article was to trace the epidemiological profile of ophidian accidents notified from 2011 to 2015 in a Toxicology Assistance Center. For this purpose, a cross-sectional research with quantitative approach was required, carried out with epidemiological data registered and documented to the notification and attention files of snakebite. Nine variables were collected for the research, and data found were analyzed by descriptive statistics. Thus, results indicate the notification of 183 events, 19.13% comprising Bothrops accidents. Male gender was the most affected, with 67,76%, and the age group from 10 to 49 was predominant (73,22%), lower limbs were the most affected body part (74,32%). The majority of the occurrences happened in rural area (54,10), mainly farmers (26,23%), as they were working (13,66%). In conclusion, it's necessary to develop researches that contribute to the (re)cognition of the epidemiological profile of such accidents as a way of favoring the development of preventive strategies, as well as to reinforce the importance of identifying the species responsible for the accidents in order to provide effective treatment and lower the risk of death occurrences and sequels to the individuals affected.

El Brasil es uno de los países sudamericanos con mayor prevalencia de accidentes ofídicos al año, sin embargo estos acontecimientos son considerados descuidados por países tropicales y subtropicales en desarrollo. El objetivo de este artículo fue trazar el perfil epidemiológico de los accidentes ofídicos notificados en el período de 2011 hasta 2015, en un Centro de Asistencia Toxicológica. Para ello, se ha hecho una investigación transversal de abordaje cuantitativo, realizado utilizando datos epidemiológicos registrados y documentados en las fichas de notificación y de atención de accidentes ofídicos. Fueron recogidas para el análisis nueve variables, y los datos encontrados fueron analizados por medio de la estadística descriptiva. Así, los resultados indicaron la notificación de 183 casos, siendo que el 19,13% comprende los accidentes botrópicos. El sexo masculino fue el más acometido, con el 67,76%, y ha predominado el grupo de edad de 10 hasta 49 años (73,22%), teniendo los miembros inferiores como la región anatómica más afectada (74,32%). La mayoría de las ocurrencias fue en la zona rural (54,10%), principalmente agricultores (26,23%), en la ocasión en que ejercían su ocupación (13,66%). Se concluyó que es necesario el desarrollo de investigaciones que contribuyan al (re)conocimiento del perfil epidemiológico de esos accidentes como forma de favorecer el desarrollo de estrategias preventivas, así como reforzar la importancia de la identificación de las especies responsables por los accidentes, para que haya un tratamiento efectivo y un menor riesgo de ocurrencia de muertes y secuelas en los individuos acometidos.

Association of oxidative stress and DNA damage with grafting time in patients with multiple myeloma and lymphoma submitted to autologous hematopoietic stem cell transplantation / Associação do estresse oxidativo e dano ao DNA com o tempo de enxertia em pacientes com mieloma múltiplo e linfomas submetidos a transplante autólogo de células-tronco hematopoéticas

ABSTRACT The aim of the study was to investigate the association between oxidative stress and DNA damage with grafting time in patients submitted to autologous hematopoietic stem-cell transplantation (HSCT). The study included 37 patients submitted to autologous HSCT diagnosed with Multiple Myeloma (MM) and lymphoma (Hodgkin’s and non-Hodgkin’s). Biomarkers of oxidative stress and DNA damage index (DI) were performed at baseline (pre-CR) of the disease and during the conditioning regimen (CR), one day after the HSCT, ten days after HSCT and twenty days after HSCT, as well as in the control group consisting of 30 healthy individuals. The outcomes showed that both groups of patients had an hyperoxidative state with high DI when compared to baseline and to the control group and that the CR exacerbated this condition. However, after the follow-up period of the study, this picture was re-established to the baseline levels of each pathology. The study patients with MM showed a mean grafting time of 10.75 days (8 to 13 days), with 10.15 days (8 to 15 days) for the lymphoma patients. In patients with MM, there was a negative correlation between the grafting time and the basal levels of GPx (r = -0.54; p = 0.034), indicating that lower levels of this important enzyme are associated with a longer grafting time. For the DI, the correlation was a positive one (r = 0.529; p = 0.030). In the group with lymphoma, it was observed that the basal levels of NOx were positively correlated with grafting time (r = 0.4664, p = 0.032). The data indicate the potential of these biomarkers as predictors of toxicity and grafting time in patients with MM and Lymphomas submitted to autologous HSCT.

RESUMO O objetivo do estudo foi investigar a associação entre estresse oxidativo e dano ao DNA com o tempo de enxertia em pacientes submetidos ao transplante de células-tronco hematopoéticas autólogo (TCTH). Participaram do estudo 37 pacientes submetidos ao TCTH autólogo com diagnóstico de mieloma múltiplo (MM) e Linfomas (Hodgkin e não Hodgkin). Biomarcadores de estresse oxidativo e índice de dano ao DNA (ID) foram determinados no estado basal (Pré-RC) das doenças e durante o regime de condicionamento (RC), um dia após o TCTH, dez dias após o TCTH e vinte dias após o TCTH e no grupo controle composto por 30 individuos saudáveis. Os resultados demonstraram que os dois grupos de pacientes apresentaram um estado hiperoxidativo com elevado ID quando comparados ao estado basal e ao grupo controle e que o RC exacerbou essa condição. No entanto, após o tempo de acompanhamento do estudo, esse quadro foi reestabelecido ao estado basal de cada patologia. Os pacientes do estudo com MM apresentaram uma média do tempo de enxertia de 10,75 dias (8 a 13 dias), e de 10,15 dias (8 a 15 dias) para o grupo Linfoma. Nos pacientes com MM houve uma correlação negativa entre o tempo de enxertia e os níveis basais de GPx (r=-0,54; p=0,034), indicando que níveis mais baixos de GPx estão relacionados a um maior tempo de enxertia, e para o ID, a correlação foi positiva (r=0,529; p=0,030). No grupo com Linfoma, observou-se que os níveis basais de NOx correlacionaram-se positivamente com o tempo de enxertia (r= 0,4664; p=0,032). Os dados apontam para o potencial desses biomarcadores como preditores da toxicidade e do tempo de enxertia em pacientes com MM e Linfomas submetidos ao TCTH autólogo

Potential chemoprotective effects of green propolis, L-lysine and celecoxib on bone marrow cells and peripheral blood lymphocytes of Wistar rats subjected to bladder chemical carcinogenesis

PURPOSE: To evaluate the genotoxicity of propolis and L-lysine, as well as their effects on the possible cellular damage in erythroblasts (bone marrow) and leukocytes (peripheral blood) caused by the carcinogen BBN (n - butyl - n {4 - hydroxybutyl} nitrosamine) in rats subjected to bladder carcinogenesis and treated with green propolis and L-lysine. METHODS: One hundred and twenty five rats were distributed into the following groups: I, IIA, IIB, III, K, L M N, X, XI, XII and XIII. Groups I to X received BBN in drinking water for 14 weeks (wks). Group I was treated with intragastric (ig) propolis at 150 mg/kg body weight, for 44 wks, beginning 30 days before start of BBN. Groups IIA and III were treated with propolis (150 mg/kg), for 40 wks, subcutaneous (sc) and ig, respectively, beginning simultaneously with BBN. On the 32nd wk, the animals of groups L, M and N were treated ig with L-lysine (300 mg/kg), celecoxib (30 mg/kg) and propolis (300 mg/kg), respectively, up to the 40th wk. The groups that received only BBN (IIB and K) were treated with water, sc and orally, respectively, for 40 wks. Groups XI, XII and XIII received respectively propolis (150 mg/kg), L-lysine (150 mg/kg) and water ig for 40 wks. After 40 wks, the surviving animals were anesthetized and subjected to femoral bone marrow aspiration and blood collection from the aorta, for CA and MNT, respectively, for investigation of genotoxicity. RESULTS: Groups IIB and K, which received only BBN and water, showed the greatest DNA damage in peripheral leukocytes (CA) and largest number of micronuclei in bone marrow erythrocytes (MNT) in relation to all other groups that received BBN and lysine and/or propolis (p<0.001). CONCLUSIONS: Both propolis and L-lysine are effective in protecting against genotoxicity, as well not being genotoxic themselves toward the cells evaluated, at the doses and times administered and according to the two tests utilized. .

Cytotoxicity and DNA damage in the neutrophils of patients with sickle cell anaemia treated with hydroxyurea

Hydroxyurea (HU) is the most important advance in the treatment of sickle cell anaemia (SCA) for preventing complications and improving quality of life for patients. However, some aspects of treatment with HU remain unclear, including their effect on and potential toxicity to other blood cells such as neutrophils. This study used the measurement of Lactate Dehydrogenase (LDH) and Methyl ThiazolTetrazolium (MTT) and the comet assay to investigate the cytotoxicity and damage index (DI) of the DNA in the neutrophils of patients with SCA using HU.In the LDH and MTT assays, a cytoprotective effect was observed in the group of patients treated, as well as an absence of toxicity. When compared to patients without the treatment, the SS group (n=20, 13 women and 07 men, aged 18-69 years), and the group of healthy individuals (AA) used as a control group (n=52, 28 women and 24 men, aged 19-60 years), The SSHU group (n=21, 11 women and 10 men, aged 19-63 years) showed a significant reduction (p<0.001) in LDH activity and an increase in the percentage of viable cells by the MTT (p<0.001). However, the SSHU group presented significantly higher DI values (49.57±6.0 U/A) when compared to the AA group (7.43 ± 0,94U/A) and the SS group (22.73 ±5.58 U/A) (p<0.0001), especially when treated for longer periods (>20 months), demonstrating that despite the cytoprotective effects in terms of cell viability, the use of HU can induce DNA damage in neutrophils.

A hidroxiuréia (HU) constitui o avanço mais importante no tratamento da anemia falciforme (AF) por prevenir complicações e aumentar a qualidade de vida dos pacientes. Entretanto, alguns aspectos do tratamento com HU permanecem obscuros, incluindo a sua ação e potencial toxicidade em outras células sanguíneas, tais como neutrófilos. Este estudo utilizou a mensuração da lactato desidrogenase (LDH) e do metil tiazoltetrazólio (MTT) e o ensaio do cometa para investigar a citotoxicidade e índice de dano (ID) ao DNA em neutrófilos de pacientes com AF em uso do medicamento. Nos ensaios de LDH e MTT, observou-se além de ausência de toxicidade, uma ação citoprotetora no grupo de pacientes tratados, Grupo SSHU (n=21, 11 mulheres e 10 homens, com idades entre 19-63 anos), quando comparados aos pacientes sem tratamento, Grupo SS (n=20, 13 mulheres e 07 homens, 18-69 anos), e grupo de indivíduos saudáveis (AA) usado como controle (n=52, 28 mulheres e 24 homens, 19-60 anos), com redução significativa (p<0,001) na atividade de LDH e aumento no percentual de células viáveis pelo MTT (p<0,001). Entretanto, o grupo SSHU apresentou valores de ID significativamente elevados (49,57±6,0 U/A), quando comparados ao grupo AA (7,43 ± 0,94U/A) e grupo SS (22,73 ±5,58 U/A) (p<0,0001), especialmente quando tratados por períodos mais longos (>20 meses), demonstrando que apesar dos efeitos citoprotetores quanto à viabilidade celular, o uso da HU pode induzir lesão ao DNA de neutrófilos.

Bioactive extracts and chemical constituents of two endophytic strains of Fusarium oxysporum

Ethyl acetate extracts of cultures grown in liquid Czapek and on solid rice media of the fungal endophyte Fusarium oxysporum SS46 isolated from the medicinal plant Smallanthus sonchifolius (Poepp.) H. Rob., Asteraceae, exhibited considerable cytotoxic activity when tested in vitro against human cancer cells. Chromatographic separation yielded anhydrofusarubin (1) and beauvericin (2) that were identified based on their ¹H and 13C NMR data. Compounds 1 and 2 showed the strongest cytotoxic activity against different cancer cell lines. Compound 2 also showed promising activity against Leishmania braziliensis. Hexanic extract of F. oxysporum SS50 grown on solid rice media also afforded a mixture of compounds that displayed cytotoxic activity against different cancer cell lines. Chemical analysis of the mixture of compounds, investigated by gas chromatography-mass spectrometry (GC-MS), showed that there was a predominance of methyl esters of fatty acids and alkanes.

Evaluating methods for the isolation of marine-derived fungal strains and production of bioactive secondary metabolites

In the present investigation we evaluate methods for the isolation and growth of marine-derived fungal strains in artificial media for the production of secondary metabolites. Inoculation of marine macroorganisms fragments in Petri dishes proved to be the most convenient procedure for the isolation of the largest number of strains. Among the growth media used, 3% malt extract showed the best result for strains isolation and growth, and yielded the largest number of strains from marine macroorganisms. The percentage of strains isolated using each of the growth media which yielded cytotoxic and/or antibiotic extracts was in the range of 23-35%, regardless of the growth media used. Further investigation of extracts obtained from different marine-derived fungal strains yielded several bioactive secondary metabolites, among which (E)-4-methoxy-5-(3-methoxybut-1-enyl)-6-methyl-2H-pyran-2-one is a new metabolite isolated from the Penicillium paxilli strain Ma(G)K.

Células-tronco na saúde e na doença / Stem cells in health and disease

Células-tronco foram recentemente caracterizadas como componentes fundamentais parao desenvolvimento e função de órgãos e tecidos do corpo humano. O descontrole nos processos de diferenciação e sobrevivência das células-tronco tem sido caracterizado como fator etiológico importante em alterações de desenvolvimento do complexo crânio-facial. O conhecimento gerado através de estudos feitos nesta área levantaram a possibilidade de se utilizar células-tronco terapeuticamente para regeneração de tecidos da cavidade oral. Por outro lado, células-tronco também foram caracterizadas em patologias importantes como o câncer. Em câncer, células-tronco participam dos processos de progressão tumoral, bem como na resistência a terapias uma vez que estas células são resistentes a quimioterapia convencional. Tanto em regeneração de tecidos quanto em câncer, é fundamental que se tenha uma compreensão profunda dos mecanismos que governam o comportamento das células-tronco para que se possa desenvolver terapias que sejam seguras e eficazes para os pacientes. O objetivo principal desta revisão é discutir o papel das células tronco em saúde e na doença, usando a regeneração de tecido pulpar e o câncer oral como exemplos de relevância no contexto de Odontologia.

Stem cells were recently characterized as critical components of the development and function of most tissues and organs in humans. Indeed, deregulated survival and differentiation of stem cells have been characterized as important etiological factors of developmental anomalies in the craniofacial region. This knowledge raised the possibility that stem cells could be useful for the therapeutic regeneration of oral tissues. On the other hand, stem cells appear to have an important role in tumor progression, as well as in the resistance of cancer to therapy. This might be due to the fact that stem cells are resilient, long-lived, and tend to survive to conventional chemotherapeutic drugs. Both, in tissue regeneration and in cancer, it is critical to have a profound understanding of the mechanisms that regulate the behavior of stem cells to develop therapies that are safe and effective for patients. The main objective of this review is to discuss the role of stem cells in health and disease, using dental pulp tissue regeneration and oral cancer as examples of relevance to Dentistry.

Pulp capping materials exert an effect on the secretion of IL-1β and IL-8 by migrating human neutrophils

Pulp repair is a complex process whose mechanisms are not yet fully understood. The first immune cells to reach the damaged pulp are neutrophils that play an important role in releasing cytokines and in phagocytosis. The objective of this study was to analyze the effect of different pulp-capping materials on the secretion of interleukin-1 beta (IL-1β) and interleukin-8 (IL-8) by migrating human neutrophils. Neutrophils were obtained from the blood of three healthy donors. The experimental groups were calcium hydroxide [Ca(OH)2], an adhesive system (Single Bond), and mineral trioxide aggregate (MTA). Untreated cells were used as control. Transwell chambers were used in performing the assays to mimic an in vivo situation of neutrophil chemotaxis. The pulp-capping materials were placed in the lower chamber and the human neutrophils, in the upper chamber. The cells were counted and the culture medium was assayed using ELISA kits for detecting and quantifying IL-1β and IL8. The data were compared by ANOVA followed by Tukey's test (p < 0.05). The secretion of IL-8 was significantly higher in all groups in comparison to the control group (p < 0.05). The adhesive system group showed higher IL-8 than the MTA group (p < 0.05). The secretion of IL-1β was significantly greater only in the MTA group (p < 0.001). It was concluded that only MTA is able to improve the secretion of IL-1β, and all materials tested increased IL-8 secretion. These results combined with all the other biological advantages of MTA indicate that it could be considered the material of choice for dental pulp capping.

Dental pulp tissue engineering

Dental pulp is a highly specialized mesenchymal tissue that has a limited regeneration capacity due to anatomical arrangement and post-mitotic nature of odontoblastic cells. Entire pulp amputation followed by pulp space disinfection and filling with an artificial material cause loss of a significant amount of dentin leaving as life-lasting sequelae a non-vital and weakened tooth. However, regenerative endodontics is an emerging field of modern tissue engineering that has demonstrated promising results using stem cells associated with scaffolds and responsive molecules. Thereby, this article reviews the most recent endeavors to regenerate pulp tissue based on tissue engineering principles and provides insightful information to readers about the different aspects involved in tissue engineering. Here, we speculate that the search for the ideal combination of cells, scaffolds, and morphogenic factors for dental pulp tissue engineering may be extended over future years and result in significant advances in other areas of dental and craniofacial research. The findings collected in this literature review show that we are now at a stage in which engineering a complex tissue, such as the dental pulp, is no longer an unachievable goal and the next decade will certainly be an exciting time for dental and craniofacial research.

A polpa dental é um tecido conjuntivo altamente especializado que possui uma restrita capacidade de regeneração, devido à sua disposição anatômica e à natureza pós-mitótica das células odontoblásticas. A remoção total da polpa, seguida da desinfecção do canal radicular e seu preenchimento com material artificial proporciona a perda de uma significante quantidade de dentina deixando como sequela um dente não vital e enfraquecido. Entretanto, a endodontia regenerativa é um campo emergente da engenharia tecidual, que demonstrou resultados promissores utilizando células-tronco associadas à scaffolds e moléculas bioativas. Desta forma, esse artigo revisa os recentes avanços obtidos na regeneração do tecido pulpar baseado nos princípios da engenharia tecidual e fornece aos leitores informações compreensivas sobre os diferentes aspectos envolvidos na engenharia tecidual. Assim, nós especulamos que a combinação ideal de células, scaffolds e moléculas bioativas pode resultar em significantes avanços em outras áreas da pesquisa odontológica. Os dados levantados em nossa revisão demonstraram que estamos em um estágio no qual, o desenvolvimento de tecidos complexos, tais como a polpa dental, não é mais inatingível e que a próxima década será um período extremamente interessante para a pesquisa odontológica.

Cytotoxicity of substances leached from a conventional and a self-etching adhesive system on human pulp fibroblasts / Citotoxicidade de substâncias liberadas de sistemas adesivos convencional e auto-condicionante sobre fibroblastos de polpa humana

The use of adhesives for direct dental pulp capping is not advisable, due to its harmful effects to the tissue. However, new adhesive systems are often released, and self-etching systems seem to be less toxic than conventional ones. The purpose of this study was to compare the in vitro cytotoxicity of substances leached from calcium hydroxide and two adhesive systems on human dental pulp fibroblasts. Cell culture media conditioned by Calcium Hydroxide (CH), Single Bond (SB), Clearfill Protect Bond primer (CP) or Clearfill Protect Bond resin (CB) were applied to human pulp fibroblasts. Fresh cell culture medium was used in the Control group. The number of viable cells was obtained through the MTT reduction assay. Data were compared by ANOVA and Tukey’s test (p≤0.05). The mean number of viable cells was 3.9x103(±0.75) for the control group, which was similar to those found in the CH group (4.31x103±0.87). Statistical differences were found among the groups (p<0.001), with the cell viability decreasing significantly with SB (0.09x103±0.06) and CP (0.28x103±0.08) when compared to CH and control groups. CB (2.37x103±0.72) was significantly less cytotoxic than CP and SB, but more cytotoxic than CH. It was concluded that Single Bond and Clearfill Protect Bond primer release substances that decrease cell viability of human dental pulp cells in culture. According to this study the use of bonding systems for direct pulp capping is not recommended, since they are cytotoxic

O uso de adesivos dentinários sobre a polpa não é recomendável, devido aos efeitos deletérios provocados sobre o tecido. Entretanto, novos sistemas adesivos são frequentemente lançados, e sistemas autocondicionantes parecem ser menos tóxicos que os sistemas convencionais. O objetivo deste estudo foi comparar in vitro, a citotoxicidade de substâncias liberadas pelo hidróxido de cálcio e dois sistemas adesivos utilizando fibroblastos de polpa humana. Meios de cultura celular condicionados por Hidróxido de cálcio (CH), Single Bond (SB), Clearfill Protect Bond primer (CP) ou Clearfill Protect Bond resin (CB) foram testados em fibroblastos de polpa humana. Meio fresco de cultivo celular foi utilizado no grupo centrole. O número de células viáveis foi obtido através do teste da redução do MTT. Os dados foram comparados por ANOVA e teste de Tukey (p≤0,05). O número médio de células viáveis foi de 3,9x103(±0,75) para o grupo controle, que foi similar ao encontrado para o grupo CH (4,31x103±0,87). Houve diferenças estatísticas entre os grupos (p<0.001), sendo que a viabilidade celular decresceu significantemente com SB (0,09x103±0,06) e CP (0,28x103±0,08), quando comparados ao CH e controle. CB (2,37x103±0,72) foi menos citotóxico que CP e SB, mas mais citotóxico que CH. Conclui-se que o Single Bond e o Clearfill Protect Bond primer liberam substâncias que diminuem a viabilidade celular de fibroblastos humanos em cultura. De acordo com este estudo, o uso de sistemas adesivos para capeamento direto não é recomendado, uma vez que são citotóxicos.

Interleukin-1 beta and interleukin-8 in healthy and inflamed dental pulps

After aggression to the dental pulp, some cells produce cytokines in order to start and control the inflammatory process. Among these cytokines, interleukin-1 beta (IL-1β) and interleukin-8 (IL-8) emerge as important ones. OBJECTIVE: The purpose of this study was to analyze the location, distribution and concentration of these cytokines in healthy and inflamed dental pulps. MATERIAL AND METHODS: Twenty pulps, obtained from healthy third molars (n=10) and from pulpectomies (n=10) were used for the study, with half of each group used for immunohistochemistry and half for protein extraction and ELISA assays. Fibroblasts obtained from healthy dental pulps, stimulated or not by Escherichia coli lipopolysaccharide (LPS), in order to simulate aggression on the cell cultures, were also used and analyzed by ELISA for IL-1β and IL-8 as complementary information. Data obtained from immunohistochemistry were qualitatively analyzed. Data obtained from ELISA assays (tissue and cells) were statistically treated by the t-test (p<0.05). RESULTS: Immunohistochemically, it was observed that inflamed pulps were strongly stained for both cytokines in inflammatory cells, while healthy pulps were not immunolabeled. ELISA from tissues quantitatively confirmed the higher presence of both cytokines. Additionally, cultured pulp fibroblasts stimulated by LPS also produce more cytokines than the control cells. CONCLUSIONS: It may be concluded that inflamed pulps present higher amounts of IL-1β and IL-8 than healthy pulps and that pulp fibroblasts stimulated by bacterial LPS produce higher levels of IL-1β and IL-8 than the control group.

Biological activity of neosergeolide and isobrucein B (and two semi-synthetic derivatives) isolated from the Amazonian medicinal plant Picrolemma sprucei (Simaroubaceae)

In the present study, in vitro techniques were used to investigate a range of biological activities of known natural quassinoids isobrucein B (1) and neosergeolide (2), known semi-synthetic derivative 1,12-diacetylisobrucein B (3), and a new semi-synthetic derivative, 12-acetylneosergeolide (4). These compounds were evaluated for general toxicity toward the brine shrimp species Artemia franciscana, cytotoxicity toward human tumour cells, larvicidal activity toward the dengue fever mosquito vector Aedes aegypti, haemolytic activity in mouse erythrocytes and antimalarial activity against the human malaria parasite Plasmodium falciparum. Compounds 1 and 2 exhibited the greatest cytotoxicity against all the tumor cells tested (IC50 = 5-27 µg/L) and against multidrug-resistant P. falciparum K1 strain (IC50 = 1.0-4.0 g/L) and 3 was only cytotoxic toward the leukaemia HL-60 strain (IC50 = 11.8 µg/L). Quassinoids 1 and 2 (LC50 = 3.2-4.4 mg/L) displayed greater lethality than derivative 4 (LC50 = 75.0 mg/L) toward A. aegypti larvae, while derivative 3 was inactive. These results suggest a novel application for these natural quassinoids as larvicides. The toxicity toward A. franciscana could be correlated with the activity in several biological models, a finding that is in agreement with the literature. Importantly, none of the studied compounds exhibited in vitro haemolytic activity, suggesting specificity of the observed cytotoxic effects. This study reveals the biological potential of quassinoids 1 and 2 and to a lesser extent their semi-synthetic derivatives for their in vitro antimalarial and cytotoxic activities.