Antinociceptive and anti-inflammatory effects of cellular and extracellular extracts from microalga Chlamydomonas pumilioniformis on mice

Microalga species have attracted interest as a source of bioactive compounds with several pharmacological activities. Previous studies reported that microalgae from the genus Chlamydomonas have anti-inflammatory and antioxidant properties. In this study, antinociceptive and anti-inflammatory activities of two extracts from microalga Chlamydomonas pumilioniformis were investigated. Cellular and extracellular extracts were prepared from a 14 day-batch culture in WC medium at the end of exponential growth and their carbohydrate contents were determined. Antinociceptive effects of extracts were evaluated by writhing and formalin-induced nociception tests, while the anti-inflammatory activity was analyzed by formalin-induced paw edema in mice. The analysis of dissolved carbohydrates detected amounts of 90 and 20 µg mL-1 of total carbohydrate in cellular and extracellular extracts, respectively. Cellular extract was mainly composed of glucose, but with significant proportions of arabinose, galactose and mannose and/or xylose and minor ones of fucose, rhamnose, amino sugars and uronic acids. Extracellular extract was composed of a similar proportion of glucose, galactose and mannose/xylose, besides significant ones of arabinose, fucose and galacturonic acid. Intraperitoneal administration of extracts significantly reduced writhing response in mice. In the formalin test, the extracellular extract inhibited both formalin phases, while the cellular extract was only effective in the late phase. Furthermore, extracts reduced the formalin-induced paw edema. In sum, we showed, for the first time, that C. pumilioniformis can be an important source of polysaccharides with anti-inflammatory and antinociceptive effects.

Associação de arcabouço de polipropileno e células tronco mesenqimais para uso em engenharia de tecido / Association of polypropylene scaffolds and mesenchymal stem cells for use in tissue engineering

A engenharia de tecidos tem como objetivo substituir tecidos danificados, manipulando células, confecção de arcabouços e a utilização de moléculas que estimulem o tecido. A proposta deste estudo foi avaliar duas técnicas de cultivo de células-tronco mesenquimais (MSC) em diferentes placas de cultura, utilizando dois tipos de telas de polipropileno (macroporoso e microporoso), para obter as melhores condições de interação entre a tela e as células, e definir uma proposta de protético para engenharia de tecidos. As telas de polipropileno foram cultivadas com células-tronco mesenquimais de tecido adiposo (ADSCs) isoladas de camundongos C57B1/6 GFP+ durante quinze dias em placas revestidas com metacrilato ou não revestidas com metacrilato. A quantidade de ADSCs aderidas foram verificadas diariamente em Câmara de Neubauer e através de uma curva de crescimento realizada pelo ensaio MTT. As ADSCs aderidas às malhas foram visualizadas com marcação de DAPI, panóticas, hematoxilina e eosina imuno-histoquímica e imunofluorescência. O melhor protocolo foi na tela microporosa, no o período de sete dias de cultivo e em placas sem metacrilato. Conclui-se que a tela de polipropileno fornece um bom suporte para as ADSCs se aderirem podendo ser utilizada na engenharia de tecidos.

Tissue engineering replaces injured tissues by manipulating cells, making scaffolds, and using molecules that stimulate the tissue. Mesenchymal stem cells (MSCs) are good candidates for tissue engineering, as this is one of the cell types which are recruited to repair injured tissues. Scaffolds are structural devices that allow cell fixation and migration, with polypropylene meshes being an example. This study aims to evaluate the culture of adipose tissue-derived mesenchymal stem cells (ADSCs), isolated from C57Bl/6 GFP + mice, in two types of polypropylene meshes (macroporous and microporous) in conventional culture plates and plates coated with methacrylate, over a period of fifteen days. The objective was to obtain the best interaction protocol between the mesh and the cells. The choice of the best method was based on adherence, maintenance of adherence and viability during culture. The amount of ADSCs adhering was checked daily by counting in a Neubauer Chamber and by using a growth curve performed with the MTT assay. The ADSCs adhering to the meshes were visualized with DAPI, panotic, hematoxylin and eosin, immunohistochemistry (integrin), and immunofluorescence (actin). ADSCs adhere to all forms of culture and to the two types of polypropylene mesh. ADSCs adhered more to the microporous mesh, within the seven day period of culture and in the plates without methacrylate. Thus, polypropylene meshes offer a good scaffold for ADSCs to adhere to.

Conus vexillum venom induces oxidative stress in Ehrlich's ascites carcinoma cells: an insight into the mechanism of induction

It is estimated that venoms of marine cone snails (genus Conus) contain more than 100,000 different small peptides with a wide range of pharmacological and biological actions. Some of these peptides were developed into potential therapeutic agents and as molecular tools to understand biological functions of nervous and cardiovascular systems. In this study we examined the cytotoxic and anticancer properties of the marine vermivorous cone snail Conus vexillum (collected from Hurgada and Sharm El-Shaikh, Red Sea, Egypt) and suggest the possible mechanisms involved. The in vitro cytotoxic effects of Conus venom were assessed against Ehrlich’s ascites carcinoma (EAC) cells. Results Conus venom treatment resulted in concentration-dependent cytotoxicity as indicated by a lactate dehydrogenase leakage assay. Apoptotic effects were measured in vivo by measuring levels of reactive oxygen species and oxidative defense agents in albino mice injected with EAC cells. Conus venom (1.25 mg/kg) induced a significant increase ( p  < 0.05) in several oxidative stress biomarkers (lipid peroxidation, protein carbonyl content and reactive nitrogen intermediates) of EAC cells after 3, 6, 9 and 12 hours of venom injection. Conus venom significantly reduced ( p  < 0.05) the activities of oxidative defense enzymes (catalase and superoxide dismutase) as well as the total antioxidant capacity of EAC cells, as evidenced by lowered levels of reduced glutathione.Conclusions These results demonstrate the cytotoxic potential of C. vexillum venom by inducing oxidative stress mediated mechanisms in tumor cells and suggest that the venom contains novel.

Development and reproductive performance of Swiss mice in an enriched environment / Desenvolvimento e performance reprodutiva de camundongos Swiss em ambiente enriquecido

The present study investigated the effects of environment enrichment on the development and reproductive performance of an outbred Swiss strain. Physical enrichment consisted of free access of animals to tubular devices of different shapes and sizes. The development evaluation was done by monitoring growth rate during 39 days, from weaning (21 days) to the onset of sexual maturity (60 days of age). Over five consecutive gestations (105 days), the following variables were monitored: litter size, number of animals born alive; number of pups alive at 12 hours and on days 4, 9, 12, 15 and 18; number of animals weaned per litter; average weight of litters on days 4, 9, 12, 15 and 18. The reproductive performance was then evaluated using the inclusion of these variables in the indices of gestation and birth, viability, lactation, survival success, mating success and production. The results showed that the environmental enrichment did not influence, in a significant way, the evaluated parameters. However, there is no reason to deprive the animals from an enriched environment where they can develop their natural instinctive behaviour and guarantee health and well-being.

O presente estudo investigou efeitos do enriquecimento ambiental sobre o desenvolvimento e a performance reprodutiva de camundongos outbred da linhagem Swiss. O enriquecimento teve como base o livre acesso dos animais a dispositivos tubulares de tamanhos e formas diferentes. A avaliação do desenvolvimento a partir da taxa de crescimento decorreu por um período de 39 dias, desde o desmame (21 dias) até os 60 dias de vida (maturidade sexual). Durante cinco gestações consecutivas (105 dias), foram registradas as seguintes variáveis: média do tamanho da ninhada, número de animais nascidos vivos; número de filhotes vivos em 12 horas, e nos dias 4, 9, 12, 15 e 18; número de animais desmamados por ninhada; e a média do peso dos filhotes nos dias 4, 9, 12, 15 e 18. A performance reprodutiva foi então avaliada a partir da inclusão dessas variáveis nos índices de gestação e nascimento, viabilidade, lactação, sucesso de sobrevivência, sucesso de acasalamento e produção. Os resultados obtidos mostraram que o enriquecimento ambiental não influenciou significativamente no desenvolvimento e na performance reprodutiva de camundongos da linhagem Swiss para nenhum dos parâmetros avaliados. Contudo, não há motivo para privar os animais de um ambiente enriquecido onde possam desenvolver seu comportamento instintivo natural e, consequentemente, viver com saúde e bem-estar.

Actividad del Flumazenil sobre el efecto depresor del Baclofén / Flumazenil acctivity on the depressing effect induced by Baclofen

Es un estudio experimental tipo ensayo terapéutico realizado en 200 ratones machos cepa NMRI, de 8 a 12 semanas de edad y peso de 31,7 gr pertenecientes al bioterio de la Uni versidad de Los Andes, con la finalidad de determinar si el Flumazenil es un antagonista competitivo o no competitivo, y su actividad sobre el efecto depresor inducido por el Baclofén mediante la evaluación de la actividad locomotora y la prueba de la barra horizontal. Previamente se estableció la dosis letal del Baclofén en 110 ratones, obteniendo una DL50 de 40,88 mg/kg para este estudio. Los resultados indican que el Flumazenil elevó la dosis letal a 47,92 mg/kg. En la prueba de la barra horizontal, prolongó el tiempo de caída de los ratones. Sobre la actividad locomotora, la administración de Baclofén produjo una reducción durante la 1ª hora (22,71%), y elevación durante la 2ª y 3ª ho ras (45,27% y 30,62% respectivamente). En el grupo trata do con Flumazenil más Baclofén se observó una reducción de la actividad locomotora en la tres horas de observación (67,47%, 60,12% y 45,85%) Se comprobó que no existe desvío de paralelismo entre las dos rectas de la curva gráfica de dosis-respuesta. Conclusión: No se conoce un antídoto efectivo para contrarrestar la depresión del SNC inducida por el Baclofén. El Flumazenil es un antagonista competitivo que actúa a nivel de los receptores gammaaminobutíricos. Se demostró en este estudio que el Flumazenil retrasó la mortalidad inducida por el Baclofén.

It is a comparative study and experimental field conducted in 100 male mice NMRI class, 8 to 12 weeks and weighing 31.7 grams of bioterio de la Universidad de Los Andes, with the aim of establishing the locomotor activity and test bar horizontal Flumazenil on the depressing effect induced by Baclofen. Before establishing the dose lethality of baclofen in 40.88 mg / kg for this study. The results indicate that Flumazenil rose 47.92 a lethal dose mg / kg. In proof of the horizontal bar, prolonged the time to drop in mice. In the group treated with Flumazenil more Baclofen was observed a reduction in activity locomotive to three hours, while Baclofen produced declined during the first hour, 1, and elevators during the 2 nd and 3 rd hour. It was found that there is no parallelism between the two lines of the dose-response curve. Conclusion: There is no known antidote to counteract the effects of the central nervous system depression induced by Baclofen. The Flumazenil is an antagonist benzodiazepines, which operates a level of receptors ganmaaminobutíricos, was demonstrated in this study decrease in mortality induced by Baclofen.

Effect of bromadiolone on haematology, liver and kidney in Mus musculus.

Bromadiolone, a second generation anticoagulant rodenticide was tested on Mus musculus to evaluate its effects on blood, liver and kidney at varied time intervals of 6, 12, 24 and 48 hrs. Groups of six animals each were selected for experiment. Animals were administered with bromadiolone in the form of bait at 6, 12, 24 and 48 hrs time intervals. Control animals were maintained for each time interval. After each time interval the experiment and the control animals were sacrificed and the effect of bromadiolone on blood, liver and kidney were studied.

Certain haematological responses in Swiss albino mice following exposure to textile dye wastewater.

Adults Swiss mice were administered 5% solution of textile industry wastewater orally for 25 days and haematological parameters like RBC, WBC, Hb, and PCV were studied. Red cell indices like MCV, MCH and MCHC were calculated. Results indicate significant reduction in RBC, Hb and PCV levels. It is inferred that toxic effluents cause metabolic alteration in erythrocytes and reduce their Hb carrying capacity.

Comparison of skin permeability of drugs in mice and human cadaver skin.

In vitro percutaneous absorption of four antihypertensive drugs were carried out across the mice and human cavader skin in order to compare their skin permeability. An interesting trend was noticed in these experiments. Poorly water soluble drug prazosin hydrochloride showed 13 times enhanced flux in the mice skin whereas the steady-state flux of the water soluble drug propranolol hydrochloride was almost same in both human cadaver and mice skin. The permeation rate of prazosin hydrochloride and propranolol hydrochloride through the human cadaver skin fluctuated widely over time, but in mice skin, distinct trends were noticed. The study indicates that the overall permeation rate in mice skin is higher than that in the cadaver skin and the meeting of the target-flux in mice skin does not guarantee its good permeability in human skin.

Evaluación del efecto de la DL-4- hidroxi, 4-etil, 4-fenil butiramida (HEPB) y la DL-3-hidroxi, 3-etil, 3-fenil propionamida (HEPP) sobre la actividad convulsiva inducida por glutamato monosódico en ratón adulto / Evaluation of the effects of DL-4-hydroxy, 4-ethyl, 4-phenyl butyramide (HEPB) and DL-3-hydroxy, 3-ethyl, 3-phenyl propionamide (HEPP) on the convulsive activity induced by monosodiumg glutamate in the adult mouse

Se ha demostrado que HEPB y HEPP son útiles anticonvulsivantes en varios modelos de convulsiones experimentales producidas en roedores. En el presente trabajo, ambos compuestos se ensayaron a dosis de 25, 50, 75 y 100 mg/kg intraperitoneal (ip) con el empleo de un modelo experimental de convulsiones mediante la inyección ip de glutamato monosódico (GMS: 5.5. mg/g) a ratones adultos. HEPB y HEPP se disolvieron en polietilenglicol-400 (PEG-400) y se inyectaron ip simultáneamente con el GMS. No obstante, que ambos fármacos no previnieron las convulsiones inducidas con GMS, hubo una clara tendencia a la reducción de la ataxia y en la intensidad de las convulsiones tónico-clónicas. El PEG-400 pareció potenciar el efecto neurocitotóxico del GMS por lo que resultó difícil evaluar adecuadamente el efecto anticonvulsivante de HEPB y HEPP diluidos en PEG-400. Se requiere realizar más estudios con el empleo de otros solventes para saber con más precisión sobre los efectos anticonvulsivantes de los fármacos en estudio, en el modelo de convulsiones inducidas con GMS en ratas y ratones

Trypanosoma cruzi infection in offspring born to chagasic C3H/He mice

This study reports the effects of Trypanosoma cruzi infection induced in C3H/He male and female mice born to chagasic mice. An experimental model was established infecting female C3H/He mice with a low virulent T. cruzi clone. In this model, mating, fertilization, pregnancy evolution and delivery was carried out successfully. The offspring was infected at four, six and eigth weeks of age. The results showed that the offspring born to chagasic mothers present decreased resistance to acquired T. cruzi infection. This decreased resistance was expressed by hiher levels of parasitaemia and higher mortality rates in offspring born to chagasic mothers than in controls. Age and sex were shown to be important factors of this phenomenon. The results suggested that maternal immune system products can modulate the immune response of the offspring.

Edema Y mionecrosis inducidos por veneno de Bothrops jararaca de Argentina en ratones / Edema and myonecrosis induced by Bothrops jararaca venom of Argentina in rats

Se estudiaron las actividades edematizante y mionecrótica inducidas por veneno de Bothrops jararaca de Argentina. Para la actividad edematizante se inocularon ratones con distintas dosis de veneno diluído en 0.05 ml de solución fisiológica. La dosis de 0.86 mug/20g ratón provocó un edema del 30 por ciento respecto al miembro opuesto en 1 hora. La evaluación de los efectos mionecróticos se realizó en ratones inoculados en el músculo gastroenemio con 70 mug de veneno em 0.1 ml de solución fisiológica, los que se sacrificaron en distinos tiempos de exprosición. Del área de inoculación se obtuvo material para procesamiento histopatológico. Los animales sacrificados a los 30 y 60 minutos revelaron edema y mionecrosis. Aquellos que se sacrificaron a las tres horas después de la inoculación, edemas de edema y hemorragia, presentaron necrosis e infiltrado inflamatorio.

Umbral del dolor a diferentes altitudes en ratones (Mus musculus)

Los misteriosos aspectos del dolor, demustran la complejidad de la definición de este fenómeno. Las áreas de gran altura representan un factor importante en la modificación de órganos, sistemas y funciones. El objetivo del presente estudio es evaluar la influencia de los factores propios de las grandes altitudes sobre el umbral del dolor. En un estudio experimental, autocontrolado, de elección deliberada; para la cual se utilizaron 50 ratones (Mus musculus), a quienes se les tomó Tiempos Análogos Umbrales del Dolor (TAUD) en Arequipa (2363 msnm), en una oportunidad (basales), y durante su exposición en Pampa Cañaguas (4142 msnm) y Mollendo (26 msnm) evaluaciones cada 8 horas durante 2 días. Se empleó el método modificado de D'Amour, cuyo estímulo térmico está representada por agua a 40ºC constante, donde el tiempo (centésimas de segundo) de permanencia de la cola del animal en el estímulo es determinada como TAUD. Tales evaluaciones fueron realizadas en laboratorios a temperatura y humedad constantes. Se obtuvo el valor basal TAUD de 1938.4 cseg. (DE_895.1). Los valores obtenidos en Pampa Cañaguas (8 Hs: X=1485.7, DE=849.1; 16 Hs: X=2757.8, DE=1380.1; 24 Hs: X= 2620.1, DE=1728.1; 32 Hs: X=3759.6, DE=1726.1; 40 Hs: X=4051.2, DE=1930.9; 48 Hs: X=3866.3, DE=1853.2) son menores en relación a los obtenidos en Mollendo (8 Hs: X=3105.2, DE=1178.2; 16 Hs: X=2336.6, DE=994.0; 24 Hs: X=2176.0, DE= 1124.5; 32 Hs: X=1988.0, DE=830.6; 40 Hs: X=2077.2, DE=853.3; 48 Hs: X=1982, DE=810.0), donde el ANOVA, en la linea de efectos principales para tiempo de exposición indica F=3.961 (P=0.0016), así mismo en la línea de efectos para grupo indica F=69.196 (P<0.0001), ambos resultados estadísticamente significativos. La interacción entre el tiempo de exposición y lugar de exposición (que refleja la altitud) es además estadísticamente importante (F=23.3; p<0.0001). Los resultados obtenidos sugieren que el umbral del dolor disminuye durante la exposición a áreas de gran altura. Se ameritan estudios posteriores, con diferentes métodos y diseños así como su estudio en seres humanos

Agregados plaquetarios en ratones inyectados con fosfotirosina / Aggregated platelets in phosphotyrosine injected mice

Reportamos la formación de agregados plaquetarios en ratones inyectados con el aminoácido fosfotirosina. Estos agregados se observaron en cortes histológicos de vasos sanguíneos, en sangre y en plasma enriquecido en plaquetas. La detección de un electrocardiograma alterado, junto a observaciones anteriores realizadas en plaquetas humanas, hacen sugerir que fosfotirosina libre podría tener un rol significativo en el proceso de activación plaquetario

Avaliaçäo farmacológica preliminar do extrato liofilizado de angico (Piptadenia colubrina Bth) / Preliminary pharmacologic evaluation of freeze-dried extract of angico (Piptadenia colubrina Bth)

Cascas de angico (Piptadenia colubrina Bth) foram coletadas na regiäo do Alto Paranaiba/Minas Gerais e submetidas a processos de extraçäo e liofilizaçäo. O extrato foi empregado em testes farmacológicos com camundongos e ratos machos albinos. Observou-se que doses variáveis do extrato provocaram sinais de depressäo do sistema nervoso central e insuficiência respiratória. Observou-se também que o extrato acarretou um efeito sinérgico quando associado a doses sub-hipnóticas de pentobarbital sódico e a doses sedativas de clorpromazina e diazepam. Embora a informaçäo popular se refira a propriedades antiepilépticas do "chá de angico", os resultados näo demonstraram açäo anticonvulsivante em relaçäo ao pentetrazol. Além do mais, observou-se que o extrato näo possui açäo analgésica e nem altera a açäo analgésica do cloridrato de morfina. Os resultados obtidos sugerem propriedades psicotrópicas do tipo psicoléptico (depressoras do sistema nervoso central) para o material estudado.