Microemulsion based on methyl 3,4,5-trihydroxybenzoate for the topical treatment of cutaneous leishmaniasis: an in vivo assay

ABSTRACT Cutaneous leishmaniasis, caused by protozoa of the genus Leishmania, presents diverse clinical manifestations, and current therapeutic options have limitations, including long treatment periods, potential hospitalization, and excessive pain during treatment. Methyl gallate, a phenolic compound found in plants such as Libidibia ferrea, presents promising antileishmanial activity. Combining this compound with existing leishmaniasis medications could lead to reduced dosages and the minimization of side effects. This study aimed to assess the efficacy of a microemulsion containing methyl gallate, either on its own or in combination with Glucantime®, for the experimental treatment of cutaneous leishmaniasis in a 30-day in vivo assay using golden hamsters infected with Leishmania (Leishmania) amazonensis. The control groups included an untreated positive control and an uninfected, untreated negative control. After treatment, we evaluated clinical, parasitological, and biochemical parameters. While none of the treatments achieved clinical or parasitological cure, notable improvements were observed in the combined group, with significant reductions in snout skin lesions and parasite load when compared to the control. Biochemical parameters such as creatinine, CK-MB, GOT, and GPT remained unchanged, but urea and CPK levels significantly increased in all the experimental groups relative to the control. In conclusion, the integration of a topical methyl gallate microemulsion with intralesional Glucantime® showed potential as an effective treatment for cutaneous leishmaniasis. Further investigations into optimal dosages and therapeutic schemes are warranted in order to enhance treatment outcomes.

RESUMO A leishmaniose cutânea, causada por protozoários do gênero Leishmania, apresenta diversas manifestações clínicas e as opções terapêuticas atuais têm limitações, incluindo longos períodos de tratamento, possível hospitalização e dor excessiva durante o tratamento. O galato de metila, um composto fenólico encontrado em plantas como Libidibia ferrea, mostra atividade antileishmania promissora. A combinação deste composto com os medicamentos existentes para leishmaniose pode levar a dosagens reduzidas e à minimização dos efeitos colaterais. Este estudo teve como objetivo avaliar a eficácia de uma microemulsão contendo galato de metila, isoladamente ou em combinação com Glucantime®, no tratamento experimental da leishmaniose cutânea em um ensaio in vivo de 30 dias em hamsters sírios infectados com Leishmania (Leishmania) amazonensis. Os grupos de controle incluíram um controle positivo não tratado e um controle negativo não infectado e não tratado. Após o tratamento, avaliamos parâmetros clínicos, parasitológicos e bioquímicos. Embora nenhum dos tratamentos tenha alcançado cura clínica ou parasitológica, melhorias notáveis ​​foram observadas no grupo combinado, com redução significativa nas lesões cutâneas do focinho e na carga parasitária em comparação com o controle. Parâmetros bioquímicos como creatinina, CK-MB, TGO e TGP permaneceram inalterados, os níveis de ureia e CPK aumentaram significativamente em todos os grupos experimentais em relação ao controle. Em conclusão, a integração da microemulsão tópica de galato de metila com Glucantime® intralesional mostrou potencial como um tratamento eficaz para a leishmaniose cutânea. Mais investigações sobre dosagens ideais e esquemas terapêuticos são necessárias para melhorar os resultados do tratamento.

Action of enfuvirtide on the pregnancy of albino rats ( Rattus norvegicus albinus, Rodentia, Mammalia ): biological assay and functional and histological analyses of exposed maternal-fetal organs

ABSTRACT Objective To assess the effects of enfuvirtide on pregnancy in albino rats and their fetuses. Methods Forty pregnant EPM 1 Wistar rats were randomly allocated into four groups: control (E) (distilled water twice/day), G1 (4mg/kg/day enfuvirtide), G2 (12mg/kg/day enfuvirtide), and G3 (36mg/kg/day enfuvirtide) groups. On the 20th day of gestation, the rats were anesthetized and subjected to cesarean section. Their blood was collected for laboratory analysis, and they were sacrificed. The offspring's fragments of their kidneys, liver, and placentas and the maternal rats' fragments of their lungs, kidneys, and liver were separated in the immediate postpartum period for light microscopy analysis. Results No maternal deaths occurred. In the second week at the end of pregnancy, the mean weight of the G3 Group was significantly lower than that of the G2 Group (p=0.029 and p=0.028, respectively). Analyzing blood laboratory parameters, the G1 Group had the lowest mean amylase level, and the G2 Group had the lowest mean hemoglobin level and the highest mean platelet count. In the morphological analysis, there were no changes in organs, such as the kidneys and liver, in both the maternal rats and offspring. Three maternal rats in the G3 Group had pulmonary inflammation in the lungs. Conclusion Enfuvirtide has no significant adverse effects on pregnancy, conceptual products, or functional alterations in maternal rats.

Atividade fotoprotetora e antioxidante de compostos fenólicos: uma revisão sistemática de testes in vitro / Photoprotective and antioxidant activity of phenolic compounds: a systematic review of in vitro tests / Actividad fotoprotectora y antioxidante de compuestos fenólicos: una revisión sistemática de ensayos in vitro

RESUMO Introdução: Os compostos fenólicos, devido a sua estrutura química, possuem a capacidade de absorver a energia ultravioleta e reduzir a formação de radicais livres. Objetivo: Avaliar a atividade fotoprotetora e antioxidante de compostos fenólicos a partir da observação de resultados in vitro e verificar a importância do uso de modelos biológicos nessa perspectiva. Metodologia: Foi realizada uma pesquisa de artigos publicados, na base de dados Pubmed, entre 2010 e 2020, que atendessem aos objetivos deste trabalho, 44 artigos foram selecionados. Resultados: Os métodos instrumentais utilizados para avaliação da atividade fotoprotetora apresentaram boa correlação in vivo e mostram-se rápidos e eficazes na determinação do fator de proteção solar. Além desses, têm-se aplicado métodos biológicos para a avaliação de aspectos que não são mensurados por métodos físico-químicos, relacionado aos danos ao DNA, decorrentes da exposição solar. Para a avaliação da atividade antioxidante, o método do radical DPPH foi empregado em 92,6 % dos estudos analisados e foi observado que os antioxidantes podem incrementar a proteção solar e, ainda, auxiliar na estabilidade de filtros solares sintéticos. Conclusão: Os compostos fenólicos, especialmente aqueles com propriedades antioxidantes, podem ser utilizados como agentes fotoprotetores em formulações tópicas para reduzir os danos à pele induzidos pela radiação UV.

SUMMARY Introduction: Phenolic compounds, due to their chemical structure, can absorb ultraviolet energy and reduce the formation of free radicals. Aim: To evaluate the photoprotective and antioxidant activity of phenolic compounds from the observation of in vitro results and to verify the importance of the use of biological models in this perspective. Methodology: A search for articles published in the Pubmed database was carried out between 2010 and 2020, which met the objectives of this work, 44 articles were selected. Results: According to the literature, the instrumental methods used to assess independent photoprotective activity, good correlation in vivo, and demonstrating rapid and effective determination of the sun protection factor. In addition to these, biological methods have been provided for the evaluation of aspects not measured by physical-chemical methods, related to DNA damage, resulting from sun exposure. For the evaluation of antioxidant activity, the DPPH radical method was registered in 92.6 % of published studies and it was observed that antioxidants can increase sun protection and also help in the stability of synthetic sunscreens. Conclusion: Phenolic compounds, especially with antioxidant properties, can be used as photoprotective agents in topical formulations to reduce skin damage induced by UV radiation.

RESUMEN Introducción: Los compuestos fenólicos, por su estructura química, tienen la capacidad de absorber la energía ultravioleta y reducir la formación de radicales libres. Objetivo: Evaluar la actividad fotoprotectora y antioxidante de compuestos fenólicos a partir de la observación de resultados in vitro y comprobar la importancia del uso de modelos biológicos en esta perspectiva. Metodología: Se realizó una búsqueda de artículos publicados en la base de datos Pubmed entre 2010 y 2020, que cumplieron con los objetivos de este trabajo, se seleccionaron 44 artículos. Resultados: Los métodos instrumentales utilizados para evaluar la actividad fotoprotectora mostraron una buena correlación in vivo y demostraron ser rápidos y eficientes en la determinación del factor de protección solar. Además de estos, se aplicaron métodos biológicos para evaluar aspectos no medidos por métodos físico-químicos, relacionados con el daño en el ADN por exposición solar. Para la evaluación de la actividad antioxidante se utilizó el método radical DPPH en el 92,6% de los estudios analizados y se observó que los antioxidantes pueden aumentar la protección solar y también ayudar en la estabilidad de los protectores solares sintéticos. Conclusión: Los compuestos fenólicos, especialmente aquellos con propiedades antioxidantes, pueden utilizarse como agentes fotoprotectores en formulaciones tópicas para reducir el daño cutáneo inducido por la radiación UV.

Cost-Effectiveness Analysis of the Implementation of Cryptococcal Antigen Lateral Flow Assay for the Diagnosis of Cryptococcal Meningitis in Symptomatic People Living With Human Immunodeficiency Virus in Brazil.

OBJECTIVES: Cryptococcal meningitis constitutes a significant source of mortality in the developing world. Annually, approximately 625 000 deaths occur worldwide among patients with human immunodeficiency virus (HIV) infection. This study aims to assess the cost-effectiveness of implementing cryptococcal antigen lateral flow assay (CRAG-LFA) screening in Brazil compared with the current practice. METHODS: An economic evaluation using a Monte Carlo microsimulation was conducted, considering the perspective of the Brazilian Public Health System, to calculate the cost-effectiveness of 4 diagnosis tests: (1) CRAG-LFA, (2) the cryptococcal antigen latex agglutination (CRAG-LA) test, (3) India ink, and (4) nontracking as a baseline. The time horizon comprised 1 year for the intervention and 5 years for the budgetary impact analysis. Two primary effectiveness outcomes were considered: years of life and quality-adjusted life-years. RESULTS: CRAG-LFA has extended dominance vis à vis CRAG-LA and India ink. CRAG-LFA would cost $418.46 more than CRAG-LA for the treatment of each symptomatic patient living with HIV, with an incremental cost effectiveness ratio of $2478.75/quality-adjusted life year. The budgetary impact analysis estimated that the incorporation of CRAG-LFA would have an additional cost of $1 959 236.50 in 5 years. CONCLUSIONS: These findings suggest that, for patients living with HIV in the Brazilian Public Health System, the adoption of CRAG-LFA screening is cost-effective compared with the use of CRAG-LA and India ink. It represents an opportunity to prevent cryptococcal meningitis and its mortality in Brazil.

Biological and physico-mechanical properties of poly (methyl methacrylate) enriched with graphene oxide as a potential biomaterial / Propiedades físicas-mecánicas-biológicas del poli (metilmetacrilato) enriquecido con óxido de grafeno como potencial biomaterial

Objective: To determine the cytotoxicity and effects of graphene oxide (GO) on cellular proliferation of gingival-fibroblasts, pulp-dental cells and human osteoblasts in culture, and to determine the physical, mechanical and biological properties of poly (methyl methacrylate) (PMMA) enriched with GO. Material and Methods: The GO was characterized with SEM. Cytotoxicity and cell proliferation were determined by the MTT bioassay. The physical mechanical tests (flexural strength and elastic modulus) were carried out with a universal testing machine. Sorption and solubility were determined by weighing before and after drying and immersion in water. Porosity was evaluated by visual inspection. Data were analyzed with Student's t-test and Tukey's posthoc ANOVA. Results: The GO has a heterogeneous morphology and a particle size of 66.67±64.76 µm. GO has a slight to no-cytotoxicity (>50-75% viability) at 1-30 days, and at 24 hours incubation of PMMA with GO significantly stimulates osteoblasts (45±8%, p<0.01). The physical and mechanical properties of PMMA with GO increase considerably without altering sorption, solubility and porosity. Conclusion: GO alone or with PMMA has an acceptable biocompatibility, could contribute to cell proliferation, cell regeneration and improve the physical mechanical properties of PMMA.

Objective: To determine the cytotoxicity and effects of graphene oxide (GO) on cellular proliferation of gingival-fibroblasts, pulpdental cells and human osteoblasts in culture, and to determine the physical, mechanical and biological properties of poly (methyl methacrylate) (PMMA) enriched with GO. Material and Methods: T he G O w as c haracterized with SEM. Cytotoxicity and cell proliferation were determined by the MTT bioassay. The physical-mechanical tests (flexural strength and elastic modulus) were carried out with a universal testing machine. Sorption and solubility were determined by weighing before and after drying and immersion in water. Porosity was evaluated by visual inspection. Data were analyzed with Student's t-test and Tukey's post-hoc ANOVA. Results: The GO has a heterogeneous morphology and a particle size of 66.67±64.76 ?m. GO has a slight to no-cytotoxicity (>50-75% viability) at 1-30 days, and at 24 hours incubation of PMMA with GO significantly stimulates osteoblasts (45±8%, p<0.01). The physical and mechanical properties of PMMA with GO increase considerably without altering sorption, solubility and porosity. Conclusion: GO alone or with PMMA has an acceptable biocompatibility, could contribute to cell proliferation, cell regeneration and improve the physical-mechanical properties of PMMA.

Biological and mechanical characterization of commercial and experimental periodontal surgical dressings

Abstract The objective of this study was to evaluate the biocompatibility and mechanical properties of two commercially available and one experimental periodontal dressing materials. The cytotoxicity of Periobond ® , Barricaid ® and one experimental periodontal dressing based on Exothane ® 8 monomer was tested on 3T3/NIH mouse fibroblast. Genotoxicity was assessed by micronuclei formation, and cell alterations were analyzed using light microscopy. Both biological assays were performed using the eluate obtained from specimens after 24, 72, or 168 hours of incubation. Mechanical characterization was assessed through the ultimate tensile strength and the water sorption and solubility tests. The significance level of α = 0.05 was used for all statistical analyses. All the materials promoted a cell viability lower than 60% in all evaluated times. In general, the cell viability was significantly reduced after 72 and 168h of specimens' incubation. Considering the factor material, there were not statistical differences in the cell viability (p = 0.156). The genotoxicity was not statistically significant among the groups in the different periods of time (p > 0.05). Differences in the ultimate tensile strength values were not statistically significant different among the groups (p = 0.125). Periobond ® showed the higher water sorption values (p < 0.001). Regarding solubility, there were no statistical differences between the groups (p = 0.098). All the periodontal dressing materials evaluated in this study exerted a cytotoxic effect against mouse fibroblasts, and their toxicity became more evident over time. Among the materials evaluated, the experimental light-cure type has shown overall similar properties to the commercial references.

A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells.

INTRODUCTION: The quantitation of glucose consumption in animal cell cultures is mainly based on the use of radiolabeled or fluorescent analogues, resulting in expensive and tedious procedures, requiring special equipment and, sometimes, with potential health and environmental risks. OBJECTIVES: The objective of this work was to evaluate the application of a blood plasma colorimetric assay to quantify glucose consumption in in vitro cultures of adipose cells. METHODS: We worked with 3T3-L1 adipose cells differentiated by 7-8 days, which were exposed to different initial glucose concentrations (5.5, 2.8 and 1.4 mM) for variable times, either in the absence or the presence of 100 nM insulin. Using a commercial colorimetric glucose assay, extracellular glucose was determined, and glucose uptake was calculated as the difference between the initial and final glucose concentration. RESULTS: The colorimetric assay allowed us to quantify glucose uptake in our cell model, observing a linear response over time (r 2 ≥0.9303) to the different glucose concentrations, both in the basal and insulin-induced condition. The insulin-stimulated glucose consumption was higher than basal consumption at all glucose concentrations evaluated, but significant differences were observed at 120-, 360- and 480-min in glucose 5.5 mM (p ≤ 0.01, n = 5), and 240 min in glucose 1.4 mM (p ≤ 0.01, n = 5). A V max of 4.1 and 5.9 nmol/ml/min (basal and insulin-induced, respectively) and a K m of 1.1 mM (same in basal vs insulin-stimulated) were calculated. The bioassay was also useful in a pharmacological context: in glucose 1.4 mM, glucose consumption showed an effect that depended on insulin concentration, with a calculated EC50 of 18.4 ± 1.1 nM. CONCLUSIONS: A simple and low-cost bioassay is proposed to quantify glucose consumption in 3T3-L1 adipose cells.

Determination of minimum bactericidal concentration, in single or combination drugs, against Mycobacterium tuberculosis.

Aim: To evaluate an assay to detect minimum bactericidal concentration (MBC) in Mycobacterium tuberculosis, using as single model rifampicin, isoniazid, levofloxacin (LVX) and linezolid (LNZ) and in combination. Material & methods: MBCs were carried out directly from resazurin microtiter assay plate and 3D checkerboard in M. tuberculosis H37Rv and five resistant clinical isolates. Results: The proposed MBC assay showed similar values to those determined by MGIT™, used as control. LVX and LNZ's MBC values were close to their MIC values. LNZ or LVX combined with isoniazid and rifampicin showed MBC value reduced in 63.7% of the assays. Conclusion: The proposed assay to determine MBCs of drugs can be applied to the study of new compounds with anti-M. tuberculosis activity to detect their bactericidal effect and also in laboratory routine for clinical dose adjustment of drugs according to the patient's profile.

TOXIC ACTIVITY OF PYRETHROIDS IN Lutzomyia longipalpis (DIPTERA: PSYCHODIDAE) FROM MAGDALENA RIVER BASIN, COLOMBIA / Actividad tóxica de piretroides en Lutzomyia longipalpis (Diptera: Psychodidae) procedente del Valle del Magdalena Medio, Colombia

ABSTRACT The study aimed to determine the toxicity of lambda-cyhalothrin, alpha-cypermethrin, and deltamethrin in L. longipalpis, through concentration-mortality bioassays. The test here was performed following WHO guidelines, but instead of using exposure WHO recipients and impregnated papers, 250 ml Wheaton glass bottles treated with 1 ml of insecticide solution were used. Batches of ten females of L. longipalpis were exposed to five concentrations of each pyrethroid that caused between 5 and 100 % mortality in this species. After 1 h of exposure, the females were transferred to observation recipients, and mortality was recorded 24 h later. The lethal concentrations (g/ml) that killed 50 and 95 % (LC50 and LC95) of the exposed L. longipalpis females were 0.05 and 0.86 for lambda-cyhalothrin, 0.24 and 3.62 for alpha-cypermethrin and 0.53 and 4.72 for deltamethrin. Based on the LC50 obtained, lambda-cyhalothrin is the most toxic pyrethroid for L. longipalpis, followed by alpha-cypermethrin and deltamethrin. It is expected that these data may be useful in studies on the effects of sub-lethal concentrations of the three pyrethroids on the behavior of L. longipalpis and studies on the vector susceptibility to these pyrethroids.

RESUMEN El objetivo del estudio fue determinar la toxicidad de los piretroides lambdacialotrina, deltametrina y alfacipermetrina en L. longipalpis, a través de ensayos concentración-mortalidad. Los ensayos se hicieron siguiendo los lineamientos de la OMS, pero en lugar de los recipientes de exposición y de los papeles impregnados de la OMS, se utilizaron botellas de vidrio Wheaton de 250 ml tratadas con 1 ml de solución de insecticida en alcohol absoluto. Grupos de 10 hembras de L. longipalpis sin alimentación sanguínea fueron expuestos a cinco concentraciones de cada piretroide, que causaron entre el 5 y 100 % de mortalidad. Pasada una hora de exposición, las hembras se trasladaron a los recipientes de observación y la mortalidad se registró 24 h después. Las concentraciones (g/ml) que mataron el 50 y el 95 % (CL50 y CL95) de las hembras expuestas de L. longipalpis fueron de 0,05 y 0,86 para la lambdacialotrina, 0,24 y 3,62 para la alfacipermetrina y 0,53 y 4,72 para la deltametrina. Basados en las CL50 obtenidas, la lambdacialotrina fue el piretroide con mayor toxicidad para L. longipalpis, seguido por la alfacipermetrina y la deltametrina. Se espera que estos datos puedan ser útiles en estudios de los efectos de concentraciones sub-letales de los tres piretroides en el comportamiento de L. longipalpis y en estudios de la susceptibilidad del vector a los mismos.

Chromosomal instability and cytotoxicity induced by ribavirin: comparative analysis in cell lines with different drug-metabolizing profiles.

Ribavirin is an important component of the treatment for hepatitis C virus (HCV) infection and, in combination with the new direct-acting antiviral (DAA) agents, comprises the major current therapeutic regimens. This study evaluated the cytotoxicity and chromosomal instability induced by ribavirin using the in vitro cytokinesis-block micronucleus cytome (CBMN-Cyt) assay in two cell lines with different expression levels of drug-metabolizing enzymes: human hepatocellular carcinoma cells (HepG2) and Chinese hamster ovary (CHO-K1) cells. HepG2 cells were treated with nine concentrations (from 15.3 µg/ml to 3.9 mg/ml) and CHO-K1 cells were exposed to eight concentrations (from 15.3 µg/ml to 1.9 mg/ml) of ribavirin for 24 h. Ribavirin inhibited cell proliferation in both cell lines, but at different concentrations: 3.9 mg/ml in HepG2 and 244.2 µg/ml in CHO-K1 cells. No significant differences were observed regarding aspects of cell death in HepG2 and CHO-K1 cells, reflecting the absence of cytotoxic effects associated to ribavirin. Ribavirin did not increase the frequency of nucleoplasmic bridges (NPBs) and nuclear bud (NBUD). However, when compared to the negative control, a significant increase in micronuclei (MNi) frequency was observed in both cell lines. However, chromosomal instability was induced by higher concentrations of ribavirin in HepG2 cells (from 61.1 to 976.8 µg/ml), compared with CHO-K1 cells (15.3 and 30.5 µg/ml). These results demonstrate the potential of ribavirin to promote chromosomal instability, and suggest that cells with different expressions of drug-metabolizing enzymes show different susceptibility to ribavirin effects.

Evaluation of Selective Physicochemical and Biological Properties of Different Root Canal Sealers.

Introduction: This in vitro study aimed to evaluate the chemical composition, water solubility, radiopacity, pH, electrical conductivity and cytotoxicity of four different root canal sealers. Methods and Materials: Four materials were tested including an epoxy resin-based sealer (AH-Plus), a calcium silicate-based sealer (MTA Fillapex), a calcium hydroxide-based sealer (Sealapex) and a zinc-oxide-eugenol-based sealer (Endofill). The materials were submitted to energy-dispersive x-ray microanalysis for elemental chemical composition. Solubility and radiopacity were evaluated according to ANSI/ADA. The pH and electrical conductivity were measured at different periods of time. L929 immortalized mouse fibroblast line were used for cytotoxicity evaluation. Statistical analyses were carried out using the ANOVA and Tukey's test. Results: The main elements were found to be silicon and calcium in MTA Fillapex, calcium and bismuth in Sealapex, zirconium and tungsten in AH-Plus and zinc and bismuth in Endofill. Sealapex had the highest value for solubility (P<0.05), AH-Plus showed the highest radiopacity value (P<0.05) while MTA Fillapex had the highest pH and electrical conductivity values (P<0.05). AH-Plus showed the highest rate of cell viability (P<0.05). Conclusion: Based on the results of this in vitro study, it was possible to conclude that Endofill and Sealpex did not meet the requirements for water solubility. The tested sealers were alkaline and showed radiopacity in accordance with ANSI/ADA standards. AH-Plus showed to be less cytotoxic than other tested root canal sealers.

O contexto atual da pesquisa clínica nas indústrias e faculdades de Medicina Veterinária do Estado de São Paulo, Brasil / The current context of clinical research in industries and schools of Veterinary Medicine in São Paulo State, Brazil

O mercado farmacêutico veterinário do Brasil é considerado um dos maiores do mundo e está em expansão. A pesquisa clínica é uma etapa importante para o desenvolvimento de produtos veterinários, no entanto, informações sobre essas atividades nas indústrias, fazendas experimentais terceirizadas e faculdades de Medicina Veterinária são escassas. Neste contexto, são descritas as atividades de pesquisa clínica nessas instituições do Estado de São Paulo. Para tanto, foram enviados questionários a representantes das indústrias farmacêuticas veterinárias e das faculdades de Medicina Veterinária do Estado. Os resultados mostraram que a maioria das indústrias tem seus escritórios na cidade de São Paulo, que em 2016 a maioria delas patrocinou entre 11 e 20 estudos, muitos deles em parceria com as faculdades. Todas as indústrias participantes já contrataram serviços de Contract Research Organization (CRO), embora relatem insatisfações com os mesmos. Por outro lado, representantes das faculdades de Medicina Veterinária declararam, em sua maioria, que conduziram dez desses estudos no ano de 2016, muitos deles com patrocínio das indústrias. Já estas se ressentem da falta de agilidade e compromisso das faculdades. A aproximação entre as indústrias de produtos veterinários e as faculdades de Medicina Veterinária foi vista pelos representantes tanto das faculdades quanto das indústrias como benéfica.

The Brazilian veterinary pharmaceutical market is considered one of the largest in the world and is in expansion. Clinical research is a crucial step in the development of veterinary products, however, information on these activities in industries, experimental farms and veterinary undergraduate courses are scarce. Given this context, this study intended to describe clinical research activities in these institutions in the state of São Paulo. For such, we sent a questionnaire to the representatives of pharmaceutical industries and institutions of veterinary medicine of the state. The results showed that the offices of most industries are located in the city of São Paulo. In 2016, most of these companies sponsored between 11 and 20 studies, many of them in partnership with departments of veterinary medicine. All industry participants contracted Contract Research Organization (CRO) services, despite reporting dissatisfaction with these services. On the other hand, most representatives from veterinary medicine courses reported that they performed 10 studies in 2016, many of them sponsored by industries. The industries resent the lack of agility and commitment of the institutions. The approximation between the veterinary industry and veterinary medicine courses was seen by representatives of both as beneficial.

O contexto atual da pesquisa clínica nas indústrias e faculdades de Medicina Veterinária do Estado de São Paulo, Brasil / The current context of clinical research in industries and schools of Veterinary Medicine in São Paulo State, Brazil

O mercado farmacêutico veterinário do Brasil é considerado um dos maiores do mundo e está em expansão. A pesquisa clínica é uma etapa importante para o desenvolvimento de produtos veterinários, no entanto, informações sobre essas atividades nas indústrias, fazendas experimentais terceirizadas e faculdades de Medicina Veterinária são escassas. Neste contexto, são descritas as atividades de pesquisa clínica nessas instituições do Estado de São Paulo. Para tanto, foram enviados questionários a representantes das indústrias farmacêuticas veterinárias e das faculdades de Medicina Veterinária do Estado. Os resultados mostraram que a maioria das indústrias tem seus escritórios na cidade de São Paulo, que em 2016 a maioria delas patrocinou entre 11 e 20 estudos, muitos deles em parceria com as faculdades. Todas as indústrias participantes já contrataram serviços de Contract Research Organization (CRO), embora relatem insatisfações com os mesmos. Por outro lado, representantes das faculdades de Medicina Veterinária declararam, em sua maioria, que conduziram dez desses estudos no ano de 2016, muitos deles com patrocínio das indústrias. Já estas se ressentem da falta de agilidade e compromisso das faculdades. A aproximação entre as indústrias de produtos veterinários e as faculdades de Medicina Veterinária foi vista pelos representantes tanto das faculdades quanto das indústrias como benéfica.(AU)

The Brazilian veterinary pharmaceutical market is considered one of the largest in the world and is in expansion. Clinical research is a crucial step in the development of veterinary products, however, information on these activities in industries, experimental farms and veterinary undergraduate courses are scarce. Given this context, this study intended to describe clinical research activities in these institutions in the state of São Paulo. For such, we sent a questionnaire to the representatives of pharmaceutical industries and institutions of veterinary medicine of the state. The results showed that the offices of most industries are located in the city of São Paulo. In 2016, most of these companies sponsored between 11 and 20 studies, many of them in partnership with departments of veterinary medicine. All industry participants contracted Contract Research Organization (CRO) services, despite reporting dissatisfaction with these services. On the other hand, most representatives from veterinary medicine courses reported that they performed 10 studies in 2016, many of them sponsored by industries. The industries resent the lack of agility and commitment of the institutions. The approximation between the veterinary industry and veterinary medicine courses was seen by representatives of both as beneficial.(AU)

Biología de Eratyrus mucronatus (Hemiptera, Reduviidae, Triatominae) en condiciones de laboratorio / Biology of Eratyrus mucronatus (Hemiptera, Reduviidae, Triatominae) in laboratory conditions

Eratyrus mucronatus, es un vector secundario de la enfermedad de Chagas con reportes de domiciliación en Colombia y Venezuela; los pocos estudios realizados muestran que en condiciones de laboratorio tiene bajos porcentajes de eclosión y altas tasas de mortalidad en su ciclo de vida. El fin de este trabajo fue establecer estadísticos vitales de esta especie para su cría en laboratorio. Se utilizó una cepa de laboratorio de E. mucronatus y se realizaron bioensayos para: estimar tiempos de desarrollo ninfal, comparar la eclosión en diferentes condiciones ambientales y comparar la fecundidad usando dos fuentes de alimentación y varios tipos de soporte. El tiempo promedio de desarrollo desde huevo hasta adulto fue de 127,6 días, con una tasa de supervivencia de 67,4% y probabilidades de morir en los estadios huevo de 0,2 y ninfa I de 0,16. Sesenta hembras semanalmente ovipusieron 744,5±120,77, con tasas de oviposición diaria por hembra de 1,19-2,69. Las tasas de eclosión más altas (80%) se obtuvieron en condiciones de temperatura de 24-25°C y humedad relativa de 65%. La fecundidad usando dos fuentes de alimentación y varios tipos de soporte, no mostró diferencias estadísticamente significativas. Se aportan datos para conocer estadísticos vitales como la duración del ciclo de vida, fecundidad y fertilidad de E. mucronatus, una especie de creciente importancia epidemiológica.

Eratyrus mucronatus is a secondary vector of Chagas disease with domicilary reports in Colombia and Venezuela. There are few studies in this specie and show under laboratory conditions low hatching rates and high mortality rates. The purpose of this study was to establish conditions that will improve the fertility of this species for laboratory rearing. A laboratory colony of E. mucronatus was used and bioassays were conducted to: estimate nymphal development times, compare hatching in different environmental conditions and compare fertility using two food sources and various types of support. The average development time from egg to adult was 127.6 days, with a survival rate of 67.4 %; egg mortality was 20% and that of nymph I 16%. 60 females produced 744.5 ± 120.77 eggs weekly, and the daily oviposition rate per female was 1.19 to 2.69. The best hatching rates (80 %) were obtained in stable conditions of temperature and relative humidity (65%). The fecundity differences using two food sources and various types of support, showed no statistical significance. The present study reports data useful for knowing vital statistics such as the cycle of life, fecundity, fertility and population dynamics of Triatominae species, especially those anthropophilic or suspected of invading the home environment, it is important to assess their potential for colonization capacity as well as in studies where the use of a large number of nymphs is required.

Cría masiva y uso de ninfas de Rhodnius pallescens, Barber, 1932 (Hemiptera, Reduviidae, Triatominae) para ensayos biológicoso / Mass breeding and use of nymph of Rhodnius pallescens, Barber, 1932 (Hemiptera, Reduviidae, Triatominae) for biological test.

Introducción: Rhodnius pallescens es una especie silvestre que hace intrusión a las viviendas en zonas en las cuales se han presentado brotes de Chagas agudo en Colombia; y para el estudio de sus características biológicas y el monitoreo de la susceptibilidad o resistencia de poblaciones de campo a insecticidas, se requiere del uso de una gran cantidad de insectos mantenidos en el laboratorio. Objetivo: Establecer condiciones de cría de ninfas de R. pallescens que permitan su mejor aprovechamiento en ensayos biológicos. Metodología: Se utilizó una cepa de laboratorio de R. pallescens proveniente de San Martin (Cesar, Colombia) y se realizaron bioensayos para: estimar el tiempo de alimentación, establecer condiciones de cría, estimar tiempos de desarrollo ninfal y comparar la fecundidad usando dos fuentes de alimentación y varios tipos de soporte. Resultados: 60 minutos de ofrecimiento de alimento permite la alimentación de 95% de las ninfas. El promedio de oviposición diario/hembra fue de 2,7 huevos y no varío significativamente con el consumo de sangre de gallina o ratón. La duración promedio del ciclo de vida desde huevo hasta el estadio ninfa-V fue de 128,6 días. El uso de cartulina negra y plumas dentro de los frascos de cría mejora la oviposición. Ninfas-V alimentadas desde ninfa-I y pesadas a los 5 o 6 días permite un aprovechamiento del 89% de las ninfas. Conclusiones: Los resultados de este trabajo brindan conocimiento para la cría masiva y el uso de ninfas de Rhodnius pallescens en ensayos biológicos.

Introduction: Rhodnius pallescens is a wild species which makes intrusion into dwellings in areas where there have been acute Chagas disease outbreaks in Colombia. Biologic research on their characteristics, and resistance to insecticides, requires the use of a large insect colony in the laboratory. Objective: To establish optimal breeding conditions of R. pallescens nymphs allowing better use in biological assays. Methodology: A laboratory strain of R. pallescens from San Martin (Cesar, Colombia) was used. Feeding time, breeding conditions, nymphal development times were assessed and fertility using two sources of feeding and different kinds of substrate was compared. Results: Providing food during 60 minutes allowed 95% of nymphs to be fed. The average daily oviposition per female was 2.7 eggs and did not vary significantly with the kind of blood used. The average duration of the life cycle from egg to nymph-V was 128.6 days. Using black cardboard and feathers in breeding jars increased oviposition. Nymphs-V (fed from nymph-I every 15 days) and weigthed at 5 or 6 days allowed the use of 89% of the nymphs. Conclusions: The results of this study allow offering recommendations for mass breeding and use of nymphs of R. pallescens in biological assays.

Evaluation of the extract of Zeyheria tuberculosa with a view to products for wound healing / Avaliação do extrato da Zeyheria tuberculosa na perspectiva de um produto para cicatrização de feridas / Evaluación del extracto de la Zeyheria tuberculosa en la perspectiva de un producto para cicatrización de heridas

OBJECTIVES: to evaluate the antimicrobial, cytotoxic and healing activities of the ethanolic extract of the stems of Z. tuberculosa via topical use and/or oral ingestion. METHOD: antimicrobial assays in vitro using the disk diffusion method, the Artemia salina toxicity test, and in vivo assays with Wistar rats. From these was collected clinical, histological and biochemical data for evaluating the healing process. RESULTS: in vitro antimicrobial testing showed activity in relation to Streptococcus pyogenes, Staphylococcus aureus and Staphylococcus epidermidis, with zones of inhibition of 18, 14 and 10 mm, respectively. The best minimum inhibitory concentration was 62.5 µg/ml for S. aureus, this bacteria being chosen for the in vitro assays. Animals treated with the ointments with the extract of Z. tuberculosa showed the best results in the reduction of the wound diameter, data confirmed by the presence of re-epithelialization in the histological samples. CONCLUSION: the extract was shown to be promising for the continuation of studies which may identify the active ingredients responsible for the pharmacological activity and its mechanism of action in the process of wound healing, so as to develop a product which may be used as an alternate means in the repair of infected cutaneous wounds. .

OBJETIVOS: avaliar as atividades antimicrobiana, citotóxica e cicatrizante do extrato etanólico do caule da Z. tuberculosa por via tópica e/ou ingestão oral. MÉTODO: ensaios antimicrobianos in vitro pelo método de difusão em disco, teste de toxicidade da Artemia salina e ensaios in vivo com ratos Wistar. Nesses foram coletados dados clínicos, histológicos e bioquímicos para avaliação do processo de cicatrização. RESULTADOS: ensaios antimicrobianos in vitro mostraram atividade frente à Streptococcus pyogenes, Staphylococcus aureus e Staphylococcus epidermidis, com halos de inibição de 18, 14 e 10mm, respectivamente. A melhor concentração inibitória mínima foi 62,5µg/mL para S. aureus, sendo essa bactéria escolhida para os ensaios in vivo. Animais tratados com as pomadas do extrato da Z. tuberculosa apresentaram melhores resultados na redução do diâmetro da ferida, dado confirmado pela presença de reepitelização nos cortes histológicos. CONCLUSÃO: o extrato mostrou-se promissor para a continuação de estudos que identifiquem os princípios ativos responsáveis pela atividade farmacológica e seu mecanismo de ação no processo de cicatrização de feridas, a fim de desenvolver um produto que possa ser utilizado de forma alternativa no reparo de feridas cutâneas infectadas. .

OBJETIVOS: evaluar las actividades antimicrobianas, citotóxicas y cicatrizantes del extracto etanólico del tallo de la Z. tuberculosa por vía tópica y/o ingestión oral. MÉTODO: ensayos antimicrobianos in vitro por el método de difusión en disco, prueba de toxicidad de la Artemia salina y ensayos in vivo con ratones Wistar. En estos fueron recolectados datos clínicos, histológicos y bioquímicos para evaluación del proceso de cicatrización. RESULTADOS: los ensayos antimicrobianos in vitro mostraron actividad frente a la Streptococcus pyogenes, Staphylococcus aureus y Staphylococcus epidermidis, con halos de inhibición de 18, 14 y 10 mm, respectivamente. La mejor concentración inhibitoria mínima fue 62,5 µg/mL para S. aureus, siendo esta bacteria escogida para los ensayos in vivo. Animales tratados con las pomadas del extracto de la Z. tuberculosa presentaron mejores resultados en la reducción del diámetro de la herida, dato confirmado por la presencia de reepitelización en los cortes histológicos. CONCLUSIÓN: el extracto se mostró promisor para la continuación de estudios que identifiquen los principios activos responsables por la actividad farmacológica y su mecanismo de acción en el proceso de cicatrización de heridas, con la finalidad de desarrollar un producto que pueda ser utilizado de forma alternativa en la reparación de heridas cutáneas infectadas. .

Composición química y capacidad anti-irritante de extractos de cuerpo entero de Ulomoides dermestoides (Coleoptera, Tenebrionidae) / Chemical composition and anti-irritant capacity of whole body extracts of Ulomoides dermestoides (Coleoptera, Tenebrionidae)

Background: In traditional medicine of Central and South America, the tenebrionid beetle Ulomoides dermestoides is used as an a phrodisiac, for the treatment of inflammatory diseases and cancer. Recently was reported cytotoxic and genotoxic properties of non-polar extract of U. dermestoides; also anti-inflammatory and immunomodulatory activity of aqueous whole body extract of beetle was reported, it suggests the existence of components with potential pharmacology use. On the other hand, it is necessary to identify those polar and non-polar extracts of U. dermestoides with anti-irritant properties for the membranes and blood vessels, which will be used in subsequence biological test and clinical assays. Objectives: The purpose of this research was to identify the chemical composition of methanolic and hexanic extracts of U. dermestoides, and to assess their anti-irritant capacity. Methods: The extracts were obtained from adult beetles of U. dermestoides. The chemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS) and the anti-irritant effect of each extract was evaluated by means of a modified assay of irritation of the chorioallantoic membrane (CAM) of fertilized chicken eggs (HET-CAM); the results were expressed as irritation index (IR). Results: Six common compounds were identified in both extracts: limonene, myristic, palmitic, estearic, oleic, and linoleic fatty acids. But in the alone methanolic extract were found: 1-pentadecanol, alpha-pinene, beta-phellandrene and alpha-terpinene, whereas in the hexanic extract were found: 2-methyl-p-benzoquinone, 2,4-dihidroxy-1-ethylbenzene, 2,5-dimethyl-quinone, saturated and unsaturated hydrocarbons and alcohols. The methanolic extract of U. dermestoides showed potential anti-irritant effect in the HET-CAM test (IR = 3.09 ± 0.11), similar to that observed with Nimesulida (IR = 2.05 ± 0.14)...

Iron bioavailability of different maize genotypes developed in a breeding program: in vitro and in vivo studies

The objective of this study was to evaluate iron bioavailability of maize genotypes, and analyze the correlation between in vitro and in vivo methods. Dialysable iron was analyzed in 13 genotypes from which 5 were selected for the biological assay. Mean iron content of the genotypes (n=13) was 17.93±2.93 mg kg-1. Phytate varied from 0.77% to 1.03%; phytate: iron molar ratio from 30.64 to 55.41; and soluble iron from 13.17 to 39.63%. The highest value for dialysable iron was 19.14%. In the biological assay, the control group, that received ferrous sulphate, did not present significant difference between the genotypes for Hb gain, Hb gain per gram of iron consumed and HRE. Hb gain did not present a significant correlation with in vitro assay. However, there were positive correlations varying from 0.653 to 0.809. The maize genotypes evaluated presented a good bioavailability since the genotypes showed the same result in hemoglobin gain than control group.

Biodisponibilidade de ferro de diferentes genótipos de milho desenvolvidos em programa de melhoramento genético: estudos in vitro e in vivo. O objetivo deste estudo foi avaliar a biodisponibilidade do ferro de genótipos de milho e analisar a correlação entre métodos in vitro e in vivo. Ferro dialisável foi analisado em13 genótipos, a partir do qual 5 foram selecionados para o ensaio biológico. A média de teor de ferro dos genótipos (n= 13) foi 17,93 ± 2,93 mg kg-1. O teor de fitato variou de 0,77% a 1,03%; razão molar fitato:ferro de 30,64 a 55,41; e ferro solúvel de 13,17 a 39,63%.O valor mais alto para o ferro dialisável foi 19,14%. No ensaio biológico, o grupo controle, que recebeu sulfato ferrso, não apresentou diferença significativa entre os genótipos no ganho Hb, ganho de Hb por grama de ferro consumido e HRE. Ganho de Hb não apresentou correlação significativa com o ensaio in vitro. No entanto, houve correlações positivas variando de 0,653 a 0,809. Os genótipos de milho avaliados apresentaram uma boa biodisponibilidade uma vez que os genótipos apresentaram o mesmo resultado quanto ao ganho de hemoglobina em relação ao grupo controle.

Comparison of the efficacy of long-lasting insecticidal nets PermaNet® 2.0 and Olyset® against Anopheles albimanus under laboratory conditions

Insecticide-treated nets provide a reduction in human-vector contact through physical barrier, mortality and/or repellent effects that protect both users and non-users, thereby protecting the wider community from vector-borne diseases like malaria. Long-lasting insecticide-treated nets (LLINs) are the best alternative. This study evaluated the bioefficacy of LLINs PermaNet® 2.0 and Olyset® under laboratory conditions with Anopheles albimanus. The laboratory strain was evaluated for insecticide susceptibility with selected insecticides used for malarial control. Regeneration time and wash resistance were evaluated with the standard bioassay cone technique following WHO guidelines. Heat assistance was used for Olyset® nets; the nets were exposed to four different temperatures to speed the regeneration process. The regeneration study of PermaNet® 2.0 showed that efficacy was fully recovered by 24 h after one and three washes and wash resistance persisted for 15 washes. Regeneration of Olyset® nets was not observed for nets washed three times, even with the different temperature exposures for up to seven days. Thus, for Olyset® the wash resistance evaluation could not proceed. Differences in response between the two LLINs may be associated with differences in manufacturing procedures and species response to the evaluated LLINs. PermaNet® 2.0 showed higher and continuous efficacy against An. albimanus.

Detección inmunoenzimática de la toxina de la bacteria Clostridium tetani: una alternativa a las pruebas biológicas con ratones / Immunoenzimatic detection of the Clostridium tetani bacterial toxin: an alternative to mice bioassays

Cell-free extracts from 20 strains of Clostridium tetani isolated from soil samples, were tested for tetanus toxin production using an enzyme immunoassay. All the extracts were classified as positive for the toxin presence, and eight of them showed absorbance values corresponding to tetanus toxin concentrations between 3.2 and 88 ng/ml; thus, they fell within the linear absorbance range (0.135-0.317). All dilutions of toxin used to obtain the calibration curve (0.0071 to 1.1 ng) were lethal for mice.