The aminoglycoside antibiotic gentamicin is able to alter metabolic activity and morphology of MDCK-C11 cells: a cell model of intercalated cells

It is well known that the aminoglycoside antibiotic gentamicin is capable of causing damage to kidney cells. Given the known involvement of Ca2+ in the nephrotoxic action of gentamicin, the purpose of this study was to establish a relationship between the concentration of intracellular Ca2+ ([Ca2+]i) and cellular cytotoxicity using MDCK-C11 cells, a clone that has several properties that resemble those of intercalated cells of the distal nephron. Changes in [Ca2+]i was determined using fluorescence microscopy. Cell viability was evaluated by the neutral red method, and cell cytotoxicity by the MTT method. The [Ca2+]i gradually increased when cells were exposed to 0.1 mM gentamicin for 10, 20, and 30 min. The presence of extracellular Ca2+ was found to be necessary to stimulate the increase in [Ca2+]i induced by gentamicin, since this stimulus disappeared by using 1.8 mM EGTA (a Ca2+ chelator). Morphological changes were observed with scanning electron microscopy in epithelial cells exposed to the antibiotic. Furthermore, with the MTT method, a decrease in metabolic activity induced by gentamicin was observed, which indicates a cytotoxic effect. In conclusion, gentamicin was able to alter [Ca2+]i, change the morphology of MDCK-C11 cells, and promote cytotoxicity.

Investigação oftalmológica pré-clínica do óleo essencial de Origanum vulgare L. Lamiaceae / Ophthalmological pre-clinical research about essential oil from the Origanum vulgare L. Lamiaceae

RESUMO Objetivo: Avaliar a irritação ocular aguda em coelhos, após a administração tópica de óleo essencial. Métodos: Para tanto, os animais foram divididos em três grupos, cada um com três coelhos, totalizando 6 olhos por grupo, e a diferença entre eles foi a concentração utilizada ( 1, 3 e 9%). Aplicou-se no saco conjuntival, de um dos olhos do animal, uma dose única de 0,1 ml do produto e o olho contralateral foi usado como controle. Analisou-se os efeitos causados pelo óleo essencial na conjuntiva, íris e córnea após 1, 24, 48, 72 horas e no final do sétimo dia após a aplicação tópica. As avaliações oftalmológicas foram feitas com o auxílio de um oftalmoscópio binocular indireto com e sem fluoresceína. As reações observadas foram graduadas segundo a escala de Draize. Foram realizados exames anatomopatológicos em todos os olhos estudados no final do experimento. Resultados: No grupo de animais submetidos à instilação ocular do óleo essencial a 1%, não se observou alterações. O tratamento com o óleo a 3% provocou alteração conjuntival no exame feito em 1 hora, o que foi reduzindo. A administração do óleo essencial a 9% induziu hiperemia conjuntival, não havendo qualquer alteração nos outros tempos de avaliação oftalmológica. Conclusão: A avaliação contribuiu para conhecer as alterações clínicas na superfície ocular. Desta forma, foi possível classificar o óleo a 1% como não irritante e nas concentrações de 3 e 9% como pouco irritante, tornando possível estudos clínicos, a fim de estabelecer o óleo como alternativa terapêutica em conjuntivites bacterianas.

ABSTRACT Objective: To evaluate acute eye irritation in rabbits following topical administration of essential oil. Methods: animals were divided into three groups, each containing three rabbits, with a total of 6 eyes per group. The difference between them was the concentration used (1, 3 and 9%). A single dose of 0.1 ml of the product was applied into the conjunctival sac of one eye of the animal, and the contralateral eye was used as control. The effects caused by the essential oil in the conjunctiva, iris and cornea were analyzed after 1, 24, 48 and 72 hours and at the end of the seventh day after topical application. Ophthalmologic evaluations were performed with the aid of a binocular indirect ophthalmoscope fluorescein and with and without the observed responses, before being graded according to the Draize scale. Pathological examinations were performed on all eyes studied at the end of the experiment. Results: in the group of animals subjected to the ocular instillation of 1% essential oil, there was no change. For treatment with 3% oil, conjunctival changes were found to be decreasing during the examination after 1 hour. Administration of the 9%essential oil induced conjunctival injection, without any change in the other ophthalmologic evaluation times. Conclusion: the evaluation contributed to meet the clinical changes in the ocular surface. Thus, it was possible to classify the oil at 1% as non-irritating and the concentration of 3% and 9 as mildly irritating, making it possible for clinical studies to establish the oil as an alternative therapy in bacterial conjunctivitis.

Estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro / Toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extract of Jatropha mollissima (Pohl) Baill (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeastern Brazil

RESUMO A cada dia, cepas bacterianas estão tornando-se resistentes a diversos antibióticos, o que faz necessária a busca de novas substâncias eficazes para o tratamento de doenças. Desta forma, este trabalho reporta o estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro. Inicialmente, realizou-se o teste de toxicidade do extrato contra Artemia salina. Na sequencia, foi realizado o ensaio antibacteriano contra quatro cepas bacterianas Gram-negativas (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883) e uma cepa Gram-positiva (Enterococcus faecalis ATCC 29212). Finalmente, fez-se a análise fitoquímica preliminar do extrato ativo para detecção das principais classes de metabólitos especiais. Como resultado, o extrato etanólico das folhas de J. mollissima se mostrou tóxico para Artemia salina, pois apresentou CL50 igual a 406,02 μg/mL. Quanto à ação antibacteriana, o extrato se mostrou ativo contra a bactéria Gram-positiva Enterococcus faecalis ATCC 29212, apresentando moderada atividade antibacteriana (halo de inibição igual a 7,03 mm). Evidenciou-se no extrato bioativo a presença de cumarinas, fenóis, taninos, flavonoides (flavonóis e flavanonas), alcaloides e esteroides, ambas as classes reportadas como antimicrobianos. Portanto, esse extrato tem potencial para ser usado na produção de fármacos contra infecções causadas por bactérias Gram-positivas. No entanto, as informações direcionam estudos futuros para o isolamento e identificação dos compostos bioativos, monitorados sob a ação antibacteriana mais expressiva.

ABSTRACT Each day, bacterial strains are becoming more resistant to various antibiotics, which requires the search for new effective substances for the treatment of diseases. Thus, this study reports the toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extracts of Jatropha mollissima (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeast of Brazil. Initially, we performed the toxicity testing of the extract against Artemia salina. Then, we conducted the antibacterial assay against four Gram-negative bacterial strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883), and one Gram-positive strain (Enterococcus faecalis ATCC 29212). Finally, we carried out the preliminary phytochemical analysis of the active extract to detect the main classes of special metabolites. As a result, the ethanolic extract of J. mollissima leaves was toxic to Artemia salina, because it presented LC50 equal to 406.02 µg/mL. Regarding antibacterial action, the extract was active against the Gram-positive bacteria Enterococcus faecalis ATCC 29212, with moderate antibacterial activity (inhibition zone equal to 7.03 mm). The bioactive extract had the presence of coumarins, phenols, tannins, flavonoids (flavanols and flavonones), alkaloids and steroids, both classes reported as antimicrobials. Therefore, this extract has the potential to be used in the production of drugs against infections caused by Gram-positive bacteria. However, these information require further studies for the isolation and identification of bioactive compounds, monitored under the more expressive antibacterial action.

Detection of Metabolites of Vecuronium Bromide in Visceral Samples Solves A Typical Death Mystery- (Toxicokinetics- Studies Using Gc-Ms Technique).

Introduction: On Sudden death of a medical student (20 year old girl) of National Institute of Medical Science autopsy materials along with site of injection were sent to F.S.L. It was informed during investigation that the girl took tetanus vaccine before death. In crime scene investigation Forensic Team observed that it was a case of gross negligence of dispensing wrong injection. Material: Tests were performed on viscera material viz., liver, spleen, kidney, lungs, brain, skin and blood etc. Mystery of suspected death was solved when a new GC-MS application was designed to get the unknown drug and various fragments of extracted material were studied. Method: A new method was developed on gas-chromatography-mass spectrophotometer and TL.C using various solvent systems is explained. Results: Metabolite fragments of vecuronium bromide a muscle relaxant were surprisingly observed in Viscera material, Blood sample and Skin piece from Leftt cubital fossa from this young girl, whose death is questioned. Conclusion: Structural elucidations of fragments provide a new approach to toxicokinetics. Th e explanation of fragments obtained were structurally compared with other neuromuscular blocking groups like atracurium and succinyl choline Th e presence of bromide attached cholest-5-en-Br, hydroxylated cholest-5-en-ol as hydroxylated product, acetylated fragments as cholest-5-enacetate and piperidone-2-one present in visceral samples indicates structural part obtained from vecuronium bromide drug. Th ese metabolites studies makes the case studies highly informative. Beside this new method of extractions, TLC systems and colouring reagents are also explained.

Recovery: revisão sistemática de um conceito / Recovery: systematic review of a concept

O conceito de recovery tem sido descrito em artigos como um estado de recuperação ou restabelecimento de funções psíquicas, físicas e sociais no funcionamento cotidiano. O objetivo do artigo é analisar concepções terminológicas em diferentes metodologias investigativas e a evolução paradigmática da noção de recovery. Pesquisa bibliográfica sistemática na base Pubmed com as palavras "recovery + schizophrenia", limitada a dois anos retrospectivos e a artigos completos gratuitos. Dezenove artigos foram analisados. A maioria destes busca associações entre dada característica e recovery, poucos são aqueles que discutem a sua concepção de forma que se distinga de termos comuns como "cura" e "reabilitação". Recovery como um estado em que o portador de transtorno mental grave possa sentir-se criador de seus caminhos tende a estar presente em estudos com metodologia qualitativa e em revisões bibliográficas, em que a medida de recovery deixa de relacionar-se à ausência de sintomas e passa a priorizar o quão participativa pode ser a vida de um indivíduo apesar da doença. Alguns estudos quantitativos vislumbram essa diferença conceitual. Em pesquisas qualitativas ocorre expansão na concepção de recovery e nas formas de promovê-lo.

The concept of recovery has been described in papers as a state of psychic, physical and social recuperation of day-to-day functions. The scope of this article is to analyze the concepts of the term in different research methodologies and the paradigmatic evolution of the recovery concept. Systematic bibliographical research was conducted in the Pubmed database using the words "recovery + schizophrenia" limited to freely available full papers published in the previous two years. Nineteen papers were analyzed. The majority of the papers sought associations between characteristic data and recovery; few papers discussed the concept in a way to distinguish it from other words like cure or rehabilitation. Recovery as a state in which people with severe mental illness can feel like the creators of their own itinerary tend to be found in qualitative studies and in bibliographic reviews in which the meaning of recovery is not related to the lack of symptoms and tends to prioritize how participative the life of an individual can be despite the disease. Some quantitative studies detect this conceptual difference. In qualitative research there is an increase in the concept of recovery and in ways of promoting it.

Análise preliminar da citotoxicidade dos aditivos alimentares urucum e cúrcuma / Cytotoxicity of the food additive annatto and turmeric

Este trabalho teve por objetivo avaliar a ação dos aditivos alimentares urucum e cúrcuma em células meristemáticas de pontas de raízes de Allium cepa nas doses de 5 e 10 g, nos tempos de exposição de 24 e 48 horas. Utilizou-se para cada dose um grupo de cinco bulbos de cebolas, que primeiramente foram enraizados em água destilada, e em seguida transferidos para as suas respectivas doses. Analisaram-se células em todo ciclo celular, totalizando 5.000 para cada controle e tempo de exposição. Os dados obtidos foram submetidos à análise estatística do Qui-quadrado (p<0,05). A partir dos resultados observou-se que as doses do urucum e do cúrcuma tiveram efeito antiproliferativo significativo sobre o ciclo celular deste sistema-teste. Também foi verificado número significativo de aberrações celulares nos dois tempos de exposição avaliados de todas as doses estudadas. Portanto, nas condições analisadas, o urucum e o cúrcuma mostraram-se citotóxicos e genotóxicos.

This study aimed to evaluate the effect of food additives annatto and turmeric tip cells of Allium cepa roots in doses of 5 or 10 g in exposure times of 24 and 48 hours. A group of five onion bulbs was used for each dose. Each dose was first embedded in distilled water and then transferred to their respective doses. Cells were analyzed throughout the cell cycle, totaling 5000 for each control and exposure time. The obtained data were subjected to statistical analysis Chi-square (p <.05). From the results it was observed that the doses of turmeric and annatto had significant antiproliferative effect on the cell cycle of this test system. They also found a significant number of cellular aberrations in the two exposure times evaluated all doses studied. Therefore, under the conditions studied, annatto and turmeric proved cytotoxic and genotoxic.

Avaliação da atividade citotóxica e potencial antiparasitário in vitro do a-pineno e carvacrol / Cytotoxic and antiparasitic in vitro activities of a-pinene and carvacrol

A leishmaniose e a doença de Chagas tem sido um grande desafio, no que diz respeito à sua terapêutica. Devido à grande dificuldade de encontrar fármacos que garantam uma ação terapêutica eficiente e menos agressora à espécie humana, diferentes produtos naturais vêm sendo testados. Muitas espécies vegetais foram investigadas quanto à sua ação leishmanicida e tripanocida na expectativa de que seus compostos metabólicos possuam atividade antiparasitária e ausência ou baixa citotoxicidade. Neste estudo sobre bioatividade do a-pineno e carvacrol, avaliaram-se os potenciais leishmanicida e tripanocida. O carvacrol apresentou um percentual de inibição de 38,34% e 74,12% para as formas promastigotas e epimastigotas respectivamente, na concentração de 100µg/mL, apresentando uma citotoxicidade de 21,62%. O a-pineno apresentou 100% e 5,30% de inibição para as formas epimastigota e promastigota na concentração de 100 µg/mL, com citotoxicidade de 87,88%.

Leishmaniasis and Chagas Disease represent a great challenge against the modern therapeutics. Due the high difficult to find new drugs with therapeutic efficacy and low toxicity, several natural products had been screened. Many species of plants were investigated about their leishmanicidal and trypanocidal activities. Some phytocompounds are the a-pinene and carvacrol. In this work, we evaluated the bioactivities of a-pinene and carvacrol against Trypanosoma cruzi and Leishmania braziliensis cell lines. The carvacrol inhibited 38,34% and 74,12% of the promatigote and epimastigote forms, respectively at 100 µg/mL, showing a low cytotoxic activity (21,62%). The O a-pinene inhibited 100% and 5,30% against the epimastigote and promastigote forms respectively, at 100 µg/mL, showing a higher cytotoxic activity (87,88%).

Early administration of peroxisomicine A1 (T-514 extracted from K. parvifolia seeds) causes necrosis of implanted TC-1 cells without affecting target organs in a murine model / La administración temprana de peroxisomicina A1 (T-514 extraída de las semillas de K. parvifolia)causa necrosis de células TC-1 implantadas sin afectar órganos blanco en un modelo murino

Peroxisomicine A1 (PA1), one of the toxins isolated from seeds of plants of the Karwinskia genus, whose targets organs are the liver, kidney, and lungs. There is a selective toxicity in vitro to cancer-cell lines derived from the lungs, liver, and colon, compared to normal cell lines. PA1 caused apoptosis in several cancer-cell lines in culture. In toxic doses to rodents, it causes extensive apoptosis in the liver, kidney, and lungs. In our study we were interested in evaluating, for the first time, the morphological effects of administration of PA1 to implanted TC-1 cells and in the target organs in vivo. The TC-1 cells were cultured and injected into the hind limb of C57BL-6 mice. The animals were divided into 3 groups; those treated with four doses of 1 mg/kg each of PA1, the untreated control, and the vehicle-control groups. All mice were killed 10 days after cell implantation. Samples were obtained from TC-1 cells at the implantation site and from the liver, kidney, and lungs. The samples were processed for examination under light and electron microscopy. In the PA1-treated group, the TC-1 cells had necrosis, whereas in the control groups the tumor cells were undamaged. The target organs did not show any lesions. We demonstrated for the first time that there is a selective toxic effect of PA1 on the TC-1 cells in vivo.

Peroxisomicina A1 (PA1), una de las toxinas aisladas de las semillas de plantas del género Karwinskia, cuyos órganos blanco son hígado, riñón y pulmón. Hay una toxicidad selectiva in vitro contra líneas celulares cancerosas derivadas de pulmón, hígado y colon, comparadas con líneas celulares normales. PA1 causa apoptosis en varias líneas celulares malignas en cultivo. En dosis tóxicas a roedores, causa extensa apoptosis en hígado, riñón y pulmón. En nuestro estudio, estuvimos interesados en evaluar por primera vez los efectos morfológicos de la administración temprana de PA1 sobre células TC-1 implantadas y los órganos blanco in vivo. Las células TC-1 fueron cultivadas e implantadas en la extremidad posterior de ratones C57BL-6. Los animales fueron divididos en tres grupos: tratado con cuatro dosis de 1 mg/kg de peso de PA1, control sin tratamiento y control vehículo. Todos los animales fueron sacrificados 10 días posterior al implante de las células. Se colectaron muestras del sitio del implante de las células TC-1 y de hígado, riñón y pulmón. Las muestras fueron procesadas para su análisis a microscopía de luz y microscopia electrónica de transmisión. En el grupo tratado con PA1, las células TC-1 presentaron necrosis, mientras que en los grupos control las células tumorales se observaron sin daño. Los órganos blanco no mostraron lesión alguna. Demostramos por primera vez que existe un efecto tóxico selectivo de PA1 sobre las células TC-1 in vivo.

Antioxidant, Antibacterial activity and Brine shrimp toxicity test of some Mountainous Lichens from Nepal

A total of twenty four lichen species belonging to six families were collected from mountainous region of Nepal. The methanol extracts of each species were tested for antimicrobial and antioxidant activitiesin vitro. It was found that extracts of twenty one lichen species were active againstB. subtilis and seven species were active againstS. aureus. Similarly, in DPPH assay, three speciesPeltigera sp.,Cladonia sp., andCanoparmelia sp. showed comparable activity with commercial standard, BHA. In ABTS+ assay, extracts ofParmoterma sp.,Ramalina sp.,Peltigera sp. andCladonia sp. showed stronger activity than ascorbic acid. The observed data after comparison with previously published reports indicated that the high altitude lichens contain stronger antioxidant and antibacterial constituents. Similarly, the methanol extracts ofHeterodermia sp. andRamalina sp. showed comparable toxicity effect with commercial standard berberine chloride indicating a potent source of anticancer drugs.

Application of liquid chromatography-high resolution mass spectrometry in toxicological screening / 法医学杂志

Due to the diversity of toxicologically relevant substances, the uncertainty of target compounds and the specificity of samples, toxicological screening techniques have always been valued by the forensic toxicologists. Depending on its powerful separation ability, superhigh resolution and accurate mass measurement, combined with the two levels spectrum database matching and abundance ratio of isotope ion, the liquid chromatography-high resolution mass spectrometry (LC-HRMS) analyzers have increasingly advantage in screening and identification of chemical compound. This review focuses on the applications of LC-HRMS in screening and identification of drug-of-abuse, prescription drugs, pesticide and stimulant. The prospect of LC-HRMS in forensic toxicology analysis is also included.

Evaluation of the acute and subchronic oral toxicity of ethanol extract from Valeriana pavonii species in Wistar rats / Evaluación de la toxicidad oral aguda y sub-crónica del extracto etanólico de la especie Valeriana pavonii en ratas Wistar

Introduction: One of the most frequent problems found in medicinal plants is the absence of clinical, toxicological, and pharmacological studies. Valeriana pavonii is one of the species used in Colombia as an anxiolytic. Further study of this specie is rendered to add information in the toxicological area. Objective: The acute and subchronic oral toxicity of V. pavonii ethanolic extract was evaluated in Wistar rats of both sexes. Materials and methods: The rats were distributed into four groups: the control group received the vehicle (0.5 mL/100 g of corporal weight) and the other three groups received increasing levels of the dosage for 90 days to evaluate characteristics like physical exam, laboratory test (blood chemistry and haematology), and anatomopathological findings. Results: This study reveals that there were no signs of toxicity, mortality, or significant alterations attributable to the ethanolic extract of V. pavonii. Conclusions: The Not Observed Adverse Effect Levels (NOAEL) of V. pavonii ethanolic extract were 2000 and 1000 mg/kg of body weight for the acute and subchronic toxicity studies, respectively.

Introducción: Uno de los problemas más frecuentes asociados con el uso de plantas medicinales es la ausencia de evidencias farmacológicas, toxicológicas y clínicas. Valeriana pavonii es una de las especies más utilizadas popularmente en Colombia con fines ansiolíticos. Es necesario avanzar en el estudio de esta especie para aportar información en el campo toxicológico. Objetivos: Evaluar la toxicidad oral aguda y sub-crónica del extracto etanólico de V. pavonii en ratas Wistar de ambos sexos. Materiales y métodos: En cada uno de los estudios se distribuyeron ratas en cuatro grupos; un grupo control que recibió únicamente vehículo (0.5 ml/100 g de peso corporal) y tres grupos correspondientes a niveles crecientes de dosis, así: para el estudio de toxicidad aguda se administraron en dosis única 20, 200 y 2000 mg/kg con un período de observación de 14 días y para el de toxicidad sub-crónica, dosis diarias de 250, 500 y 1000 mg/kg durante 90 días. Se evaluaron los parámetros de examen físico, los exámenes de laboratorio (química sanguínea y hematología) y el estudio anatomopatológico. Resultados: No se presentaron signos de toxicidad, letalidad ni alteraciones significativas atribuibles al consumo del extracto etanólico de V. pavonii, según el examen físico, el examen anatomopatológico y el análisis de las pruebas de química sanguínea y hematología. Conclusiones: Los valores de nivel sin efectos adversos observados (NOAEL) del extracto etanólico de V. pavonii, fueron 2000 y 1000 mg/kg de peso corporal para los estudios de toxicidad aguda y sub-crónica, respectivamente. No se encontraron valores de nivel más bajo de efecto adverso observado (LOAEL).

Aspectos clínicos da intoxicação experimental pelas favas de Stryphnodendron fissuratum (Leg. Mimosoideae) em caprinos / Clinical aspects of the experimental poisoning by the pods of Stryphnodendron fissuratum (Leg. Mimosoideae) in goats

Com o objetivo de caracterizar o quadro clínico da intoxicação por Stryphnodendron fissuratum Mart. (Leg. Mimosoideae) em caprinos, administraram-se as favas dessa planta a oito caprinos, por via oral forçada em doses únicas e a outros dois caprinos, em doses fracionadas. A menor dose que causou sinais clínicos e morte foi a de 10g/kg. Doses de 20g/kg e 40g/kg causaram sinais acentuados e doses únicas de 5g/kg não provocaram sinais. Doses fracionadas de 5g/kg durante quatro dias, totalizando 20g/kg provocaram sinais acentuados e morte. Em ambos os grupos, os primeiros sinais de intoxicação foram observados a partir do primeiro dia de experimento e a evolução variou de 4-25 dias. A doença caracterizou-se principalmente por alterações digestórias e nervosas que consistiram em anorexia, desidratação, hipomotilidade e atonia ruminal, timpanismo, gemidos constantes, dor à percussão abdominal, fezes com muco, ranger de dentes, apatia, ataxia, dismetria, tremores de cabeça, tremores musculares, fraqueza com o andar cambaleante e trôpego, acentuada depressão e decúbito esternal ou lateral prolongado e morte. Alguns animais apresentaram acentuada queda de pêlos na região dorsal; apenas um caprino apresentou fezes líquidas, marrom-escuras e fétidas. Outros sinais incluíram perda de fluido ruminal durante a ruminação, sialorréia, exsudato nasal seroso e lacrimejamento. As provas de função hepática e renal revelaram alterações discretas. As concentrações séricas de aspartato aminotransferase encontraram-se levemente aumentadas e as de creatinofosfocinase muito aumentadas.

In order to confirm the susceptibility of goats to the poisoning by Stryphnodendron fissuratum Mart. (Leg. Mimosoideae) and to characterize the clinical disease, the pods of the plant were given orally to each of eight young goats and in fractioned doses to other two. The lowest lethal dose was 10g/kg. The same dose was the lowest that induced disease. Doses of 20g/kg and 40g/kg caused pronounced clinical signs and doses of 5g/kg did not caused signs. Fractioned doses of 5g/kg during four days also caused pronounced signs. In each groups the first signs of poisoning were observed from the first day of experiments and the changes ranged from 4-25 days. The disease was characterized mainly by digestive and nervous disorders. Clinical signs were partial to complete anorexia, dehydration, decrease in ruminal activity up to atonia, tympanism, constant vocalizations, grinding of the teeth pain up on abdominal palpation, apathy, ataxia, depression, dysmetria, head and muscle tremors, weakness, difficulty in rising, sternal or lateral recumbency and death. Some goats presented extense hair loss in the skin of the dorsum; one goat presented liquid and black fetid feces. Other signs included loss of ruminal fluid during rumination, drooling, serous nasal and ocular discharges. Liver and kidney function tests had resulted in slight changes. AST serum levels were slightly increased and creatine phosphokinase levels were highly increased. These changes can associated to the effects of triterpenic saponins contained in the S. fissuratum pods.

Aspectos clínicos da intoxicação experimental pelas favas de Stryphnodendron fissuratum (Leg. Mimosoideae) em caprinos / Clinical aspects of the experimental poisoning by the pods of Stryphnodendron fissuratum (Leg. Mimosoideae) in goats

Com o objetivo de caracterizar o quadro clínico da intoxicação por Stryphnodendron fissuratum Mart. (Leg. Mimosoideae) em caprinos, administraram-se as favas dessa planta a oito caprinos, por via oral forçada em doses únicas e a outros dois caprinos, em doses fracionadas. A menor dose que causou sinais clínicos e morte foi a de 10g/kg. Doses de 20g/kg e 40g/kg causaram sinais acentuados e doses únicas de 5g/kg não provocaram sinais. Doses fracionadas de 5g/kg durante quatro dias, totalizando 20g/kg provocaram sinais acentuados e morte. Em ambos os grupos, os primeiros sinais de intoxicação foram observados a partir do primeiro dia de experimento e a evolução variou de 4-25 dias. A doença caracterizou-se principalmente por alterações digestórias e nervosas que consistiram em anorexia, desidratação, hipomotilidade e atonia ruminal, timpanismo, gemidos constantes, dor à percussão abdominal, fezes com muco, ranger de dentes, apatia, ataxia, dismetria, tremores de cabeça, tremores musculares, fraqueza com o andar cambaleante e trôpego, acentuada depressão e decúbito esternal ou lateral prolongado e morte. Alguns animais apresentaram acentuada queda de pêlos na região dorsal; apenas um caprino apresentou fezes líquidas, marrom-escuras e fétidas. Outros sinais incluíram perda de fluido ruminal durante a ruminação, sialorréia, exsudato nasal seroso e lacrimejamento. As provas de função hepática e renal revelaram alterações discretas. As concentrações séricas de aspartato aminotransferase encontraram-se levemente aumentadas e as de creatinofosfocinase muito aumentadas.

In order to confirm the susceptibility of goats to the poisoning by Stryphnodendron fissuratum Mart. (Leg. Mimosoideae) and to characterize the clinical disease, the pods of the plant were given orally to each of eight young goats and in fractioned doses to other two. The lowest lethal dose was 10g/kg. The same dose was the lowest that induced disease. Doses of 20g/kg and 40g/kg caused pronounced clinical signs and doses of 5g/kg did not caused signs. Fractioned doses of 5g/kg during four days also caused pronounced signs. In each groups the first signs of poisoning were observed from the first day of experiments and the changes ranged from 4-25 days. The disease was characterized mainly by digestive and nervous disorders. Clinical signs were partial to complete anorexia, dehydration, decrease in ruminal activity up to atonia, tympanism, constant vocalizations, grinding of the teeth pain up on abdominal palpation, apathy, ataxia, depression, dysmetria, head and muscle tremors, weakness, difficulty in rising, sternal or lateral recumbency and death. Some goats presented extense hair loss in the skin of the dorsum; one goat presented liquid and black fetid feces. Other signs included loss of ruminal fluid during rumination, drooling, serous nasal and ocular discharges. Liver and kidney function tests had resulted in slight changes. AST serum levels were slightly increased and creatine phosphokinase levels were highly increased. These changes can associated to the effects of triterpenic saponins contained in the S. fissuratum pods.

Corrositex®, BCOP and HET-CAM as alternative methods to animal experimentation / Corrositex®, BCOP e HET-CAM como métodos alternativos à experimentação animal

Tests in animals are used as models in toxicological and investigative studies. However, such tests have been considered inhumane because they can cause pain and suffering to experimental animals, while these methods can often be subjective. Protests calling for animal protection have questioned the effectiveness of in vivo tests and suggest the introduction of alternative, in vitro methods. International organizations, such as the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), the National Institute of Health (NIH), the Organization for Economic Co-operation and Development (OECD), that regulate and develop new alternative animal models, have indicated the running of preliminary assays and execution of sequential tests, which consider physical-chemical properties and data of in vitro assays, before performing in vivo studies. Towards this background, the objective of the present article was to select promising alternative methods such as Corrositex®, BCOP and HET-CAM, intended to refine or replace the use of animals and reduce their suffering.

Testes em animais são utilizados como modelos em estudos toxicológicos e de pesquisa. Entretanto, tais testes têm sido considerados desumanos, porque causam dor e sofrimento aos animais experimentais, porquanto estes métodos podem, freqüentemente, ser subjetivos. Protestos clamando pela proteção animal têm questionado a eficácia dos testes in vivo e sugerem a introdução de métodos alternativos in vitro. Organizações internacionais, tais como Comitê de Coordenação Interagências de Métodos de Validação Alternativos (ICCVAM), Instituto Nacional de Saúde (NIH), Organização para Cooperação Econômica e Desenvolvimento (OECD), que regulam e desenvolvem novos métodos alternativos aos modelos animais, indicaram a realização de ensaios preliminares e a execução de testes seqüenciais, que consideram as propriedades físico-químicas e os dados dos ensaios in vitro, antes de efetuarem estudos in vivo. Nessa direção, o objetivo do presente artigo foi selecionar métodos alternativos promissores, tais como Corrositex®, BCOP e HET-CAM, com o intuito de aperfeiçoar ou substituir o uso de animais e reduzir seus sofrimento.

Reproduction of Cubaris murina (Crustacea: Isopoda) under laboratory conditions and its use in ecotoxicity tests / Reprodução de Cubaris murina (Crustacea: Isopoda) em condições de laboratório e seu uso em testes de ecotoxicidade

Reproduction of the isopod Cubaris murina was studied in the laboratory in order to observe the offspring number per brood, the number and duration of the incubation periods, and the influence of repeated mating on number of young per brood. An experiment was conducted with two different groups: female/male pairs and females that were isolated after mating. No significant difference was found between the two reproductive female groups for the mean number of young per brood (x = 25) and the duration of the incubation periods (x = 17 days). One mating was enough for at least five broods, with no reduction in brood size. There was no significant difference between the number of adult males and females. Low-density cultures had a significantly higher growth rate compared to high-density cultures. Furthermore, the experimental results for optimization isopod culture conditions for terrestrial ecotoxicity testing are reported.

A reprodução do isópodo Cubaris murina foi investigada em laboratório para observar: o número de filhotes por ninhada, número e duração dos períodos de incubação, e se repetidas cópulas influenciam o número de juvenis por ninhada. Um experimento foi conduzido com dois diferentes grupos: pares com uma fêmea e um macho e fêmeas mantidas individualmente depois da cópula. Não foram encontradas diferenças estatisticamente significantes entre os dois grupos de fêmeas em relação ao número de filhotes por ninhada (x = 25) e a duração dos períodos de incubação (x = 17 dias). Uma cópula foi suficiente para ao menos cinco ninhadas, sem redução em seu tamanho. A proporção sexual dos adultos de C. murina em laboratório não apresentou diferenças significativas entre a proporção de machos e fêmeas nos cultivos. A densidade influencia o crescimento dos isópodos de forma significativa: culturas com baixa densidade têm maior taxa de crescimento do que culturas com alta densidade. Os resultados experimentais sobre a eficiência dos cultivos em laboratório para ensaios ecotoxicológicos terrestres são apresentados.

Níveis elevados de manganês e déficit cognitivo em crianças residentes nas proximidades de uma metalúrgica ferro-manganês na região metropolitana de Salvador, Bahia / Elevated levels of manganese and cognitive impairment in children living near a metallurgical iron-manganese in the Metropolitan Region of Salvador, Bahia

As crianças, sobretudo aquelas socialmente vulneráveis, são mais susceptíveis aos efeitos tóxicos da exposição ambiental aos agentes químicos. No processo de desenvolvimento, o sistema nervoso imaturo apresenta grande oportunidade de ação de contaminantes ambientais como o mercúrio (Hg), chumbo (Pb) e o manganês (Mn). Os objetivos desta investigação foram quantificar o grau de exposição ao Mn em crianças residentes nas proximidades de uma planta metalúrgica de ligas ferro-manganês e avaliar a associação entre os níveis deste metal no sangue e no cabelo e efeitos na função cognitiva. Para tal fim, foi realizada uma revisão da literatura científica sobre exposição de crianças ao Mn e efeitos neuropsicológicos, a qual originou o primeiro artigo. A avaliação da exposição ao Mn foi realizada na Vila Cotegipe, no município de Simões Filho, Bahia. Na primeira etapa do trabalho foram obtidas das crianças de 1 a 10 anos amostras de cabelo para determinação do Mn, sangue para hemograma e amostras para parasitológico de fezes. Foram também coletadas amostras ambientais como: água bruta e tratada, material particulado na fração respirável (PM2.5) e poeira domiciliar. Para fins de comparação, crianças de uma comunidade distante 7,5 km da metalúrgica, e a favor dos ventos, foram incluídas como grupo controle Na segunda etapa da avaliação, foram incluídas somente as crianças de 6 a 11 anos e 11 meses matriculadas na escola municipal local. Novas amostras de cabelo e sangue foram coletadas para análise de Mn, chumbo (Pb) e ferro sérico, sendo solicitada a mãe ou responsável a doação da amostra de cabelo. Nessa etapa foi realizada a avaliação cognitiva, através dos instrumentos WISC-III (Wechesler Intelligence Scale for Children), matriz progressiva de Raven para medir a cognição materna e inventário HOME adaptado para estimar o ambiente familiar...

Children, especially those socially vulnerable, are more susceptible to toxic effects resulting from environmental exposure to chemical agents. The developing nervous system has great opportunities to the action of environmental contaminants like mercury (Hg), lead (Pb) and manganese (Mn). The objectives of this research were to evaluate the Mn exposure levels in children living in the vicinity of a ferro-manganese alloy plant and investigate the association between Mn levels in blood and hair with the effects on the cognitive function. Initially, we carried out an intensive literature review on the association between children's exposure to Mn and neuropsychological effects, which led to the first article. The field work started with the populational pool and registration of all families within the limits of the Cotegipe Village, Simões Filho town, Bahia, Brazil. After obtaining the informed consent, we collected socio-demographic data among the volunteers. To assess Mn exposure level, we performed the first sampling campaign with children aged 1 to 10 years: hair samples for Mn determination, blood sample for haemogram and stool for intestinal parasites analyses. At this phase we collected environmental samples: water pre a post treatment, particulate matter from respirable fraction (PM2.5) and house dust. In the second exposure assessment campaign we included only children aged 6 to 11 years and 11 months, enrolled in the local public school, who provided hair and blood samples for Mn, lead and serum iron determination. Mothers or caregivers were asked to provide hair sample. This happened concomitantly with the cognitive evaluation, which was assessed using WISC-III (Wechesler Intelligence Scale for Children), Raven's Progressive Matrices for measuring maternal cognition and the adapted HOME to estimate the family environment stimulation...

A simple method to measure cell viability in proliferation and cytotoxicity assays

Resazurin dye has been broadly used as indicator of cell viability in several types of assays for evaluation of the biocompatibility of medical and dental materials. Mitochondrial enzymes, as carriers of diaphorase activities, are probably responsible for the transference of electrons from NADPH + H+ to resazurin, which is reduced to resorufin. The level of reduction can be quantified by spectrophotometers since resazurin exhibits an absorption peak at 600 çm and resorufin at 570 çm wavelengths. However, the requirement of a spectrophotometer and specific filters for the quantification could be a barrier to many laboratories. Digital cameras containing red, green and blue filters, which allow the capture of red (600 to 700 çm) and green (500 to 600 çm) light wavelengths in ranges bordering on resazurin and resorufin absorption bands, could be used as an alternative method for the assessment of resazurin and resorufin concentrations. Thus, our aim was to develop a simple, cheap and precise method based on a digital CCD camera to measure the reduction of resazurin. We compared the capability of the CCD-based method to distinguish different concentrations of L929 and normal Human buccal fibroblast cell lines with that of a conventional microplate reader. The correlation was analyzed through the Pearson coefficient. The results showed a strong association between the measurements of the method developed here and those made with the microplate reader (r² = 0.996; p < 0.01) and with the cellular concentrations (r² = 0.965; p < 0.01). We concluded that the developed Colorimetric Quantification System based on CCD Images allowed rapid assessment of the cultured cell concentrations with simple equipment at a reduced cost.

Agrotóxicos e saúde: realidade e desafios para mudança de práticas na agricultura / Pesticides and health: reality and challenges for change of practices in agriculture

Este artigo apresenta os resultados de uma pesquisa que teve como objetivo principal caracterizar o contexto e as práticas relacionadas ao uso de agrotóxicos entre agricultores residentes na Serrinha do Mendanha, comunidade agrícola situada em Campo Grande, município do Rio de Janeiro. A proposta metodológica teve uma abordagem qualitativa, sendo a principal fonte de dados a aplicação de questionário semiestruturado. Participaram da pesquisa 38 agricultores familiares. O perfil socioeconômico mostra uma realidade de idosos no trabalho agrícola, percentuais expressivos de analfabetismo e baixa renda. O uso de agrotóxicos é uma rotina para essas famílias. Apesar de existir uma naturalização do uso, a maioria dos informantes acredita que agrotóxicos podem afetar sua saúde, apresenta preocupação com o consumidor e gostaria de conhecer formas alternativas de cultivo. Tais achados apontam para a possibilidade de busca de mudança de práticas. Não se trata de repasse de conhecimento, pois este tipo de informação de alguma forma eles já possuem. Trata-se de desenvolver estratégias objetivando a minimização dos riscos ou mesmo o desuso de agrotóxicos na lavoura.

This paper presents the results of a research which main goal was to describe the context and practices related to the use of pesticides among farmers in the community of Serrinha do Mendanha, Campo Grande, County of Rio de Janeiro. The methodological proposal had a qualitative approach and data was collected through a semi-structured questionnaire. Thirty eight farmers participated in this research. Socio-economic profile shows elderly people working in agriculture, expressive percentile of illiteracy and low income. Despite the natural approach to the use of pesticides, which is a routine to these families, most informants believe that pesticides can affect their health, show some concern about the consumers and would like to know alternatives. These findings point to the possibility of change in agricultural practices. It is not a matter of simply offering information that in some way they already possess; it means developing strategies to reduce or even quit the use of pesticides in plantations.

Mercúrio total em cabelo de crianças de uma população costeira, Cananéia, São Paulo, Brasil / Total hair mercury in children from a coastal population in Cananéia, São Paulo State, Brazil

Os níveis de mercúrio (Hg) total em cabelos estão diretamente relacionados à alimentação, particularmente ao consumo de peixes por populações costeiras com grande representação caiçara. No presente estudo foram avaliados os níveis de mercúrio total em cabelos de crianças com idade entre 4 e 12 anos, pertencente a três escolas públicas da cidade de Cananéia, São Paulo, Brasil. Os resultados obtidos (mediana e intervalo) para mercúrio total foram de: 0,04mg.kg-1 (0,01-0,77mg.kg-1), 0,39mg.kg-1 (< 0,01-3,33mg.kg-1) e 0,39mg.kg-1 (< 0,01-2,81mg.kg-1) considerando as escolas ES1, ES2 e ES3, respectivamente. Em geral, os valores encontrados estiveram bem abaixo do valor preconizado pela Organização Mundial da Saúde para uma população adulta não exposta ao mercúrio (2,0mg.kg-1). Os baixos valores observados e a inexistência de valores de referência para mercúrio total em cabelos de crianças brasileiras possibilitam a consideração desses valores como possível referência nacional em cabelos de populações costeiras, uma vez que foram obtidos em região de baixo impacto ambiental.

Mercury (Hg) levels in hair are directly related to eating habits, especially fish consumption by coastal populations with a large contingent of traditional fishing families. This study assessed total Hg levels in children's hair. The study group was selected from three public elementary schools in Cananéia, São Paulo State, Brazil (ages 4 to 12 years). The results (median and range) for total Hg levels in children's hair were: 0.04mg.kg-1 (0.01-0.77mg.kg-1), 0.39mg.kg-1 (< 0.01-3.33mg.kg-1), and 0.39mg.kg-1 (< 0.01-2.81mg.kg-1) for schools ES1, ES2, and ES3, respectively. The values were well below the level set by World Health Organization for an adult population unexposed to Hg (2.0mg.kg-1). However, since there are no existing reference values for total Hg in children's hair, these results can be used as a contribution to establishing reference values for total hair Hg in Brazilian children living in coastal areas.