Antiproliferative activities of Althaea ludwigii L. extract on Michigan Cancer Foundation-7 breast cancer cell line

Cancer has been considered one of these major community healthiness problems. Althaea ludwigii L.(family: Malvaceae)is an indigenous plant, widely distributed in Iraq. The objective of this study is to explore the antiproliferative action ofA. ludwigii L. against breast cancer cell line Michigan Cancer Foundation-7 (MCF-7) and rate embryonic fibroblasts(REF) normal cell line and to identify the bioactive constituent. The aerial parts of A. ludwigii L. were grinded intofine powder then extracted with 85% methanol. Methanol extract was further partitioned with ethyl acetate. Ethylacetate fraction was chosen to test against MCF-7 by (3-[4, 5-dimethylthiazol-2-yl]-2, 5 (MTT assay) the bioactiveconstituents were investigated by high performance thin layer chromatography high-performance thin-layerchromatography (HPTLC). The data showed that ethyl acetate fraction has significant antiproliferative activity againstMCF-7 (IC50 = 35.5 µg/ml) and REF (IC50 = 1762.9 µg/ml), HPTLC showed the existence of rutin in ethyl acetatefraction. The antiproliferative activity may relate to the existence of rutin which had been approved as potent anticancer.

Cytotoxicity and antiviral activity evaluation of Cymbopogon spp hydroethanolic extracts

Cymbopogon citratus and C. nardus are noteworthy among the several existing plant species displaying medicinal properties, due to the potential pharmacological activity of these species, including antiviral, antibacterial, antifungal and anti-trypanosomal activities. The objective of this study was to carry out in vitro toxicity tests of plant extracts from both species and analyze potential antiviral activity against Human mastadenovirus serotype 5 (HAdV-5). Two cell lines (A549 and VERO) were used and mitochondrial and lysosomal viability were determined by the MTT and neutral red assay, respectively, after two exposure times (24 hours and six days). The aim of these assays was to counteract the behavior of the extracts against the different cell lines and determine their non-toxic concentration range, in order to evaluate possible antiviral activity against HAdV-5. Plaque reduction and inhibition index of viral titer assays were performed using the maximum non-cytotoxic concentrations (MNCC) of each extract. The results indicate MNCC at 625 µg/mL for all extracts, except for Cymbopogon nardus obtained with 80% ethanol (CN80), which showed toxicity at concentrations higher than 312.5 µg/mL. CN80 was the only extract that displayed potential activity against HAdV-5, at a concentration of 75 µg/mL, becoming a candidate for extract fraction purification and/or the isolation of substances related to the observed antiviral activity

Euphorbia umbellata bark extracts – an in vitro cytotoxic study

ABSTRACT Euphorbia umbellata (Pax) Bruyns, Euphorbiaceae, is commonly used in folk medicine of southern Brazil to treat several kinds of cancer. The latex (part of the plant used for this purpose) is mixed with water and taken as treatment; but this matrix contains toxic potential related to the presence of some phorbol type diterpenes. So the aim of this study was to evaluate the cytotoxicity of the crude extract of the bark of E. umbellata and its fractions (Hex, CHCl3, EtOAc and MeOH) using in vitro assay (applying Jurkat cells line). A preliminary cytotoxic study (MTT reduction, trypan blue exclusion and DNA quantification assays) was executed to identify the most active material. The CHCl3 fraction displayed the highest activity and was selected for further investigation of any cytotoxic mechanism and evaluation of chemical composition; flow cytometry, Acridine orange and Hoechst 33342 staining experiments and Gas chromatography–mass spectrometry analysis were applied to achieve these results. This fraction demonstrated the best cytotoxic results against Jurkat cells line with IC50 of 29.00 ± 1.49, 10.06 ± 1.48 and 4.83 ± 2.25 µg/ml for 24, 48 and 72 h of experiment, respectively (trypan blue exclusion). The mechanism responsible for this action can be associated with the promotion of cell cycle arrest and apoptosis. The two main classes of compounds present in the CHCl3 fraction are steroids and triterpenes. Further, phytochemical studies with this fraction need to be evaluated, to try isolating these substances and establishing a more detailed cytotoxic study against Jurkat cells.

Comparação de dois métodos de colheita de medula óssea de equinos e de dois meios de diluição para o isolamento de células mononucleares / Comparison of two methods of equine bone marrow aspiration and two solutions for mononuclear cell isolation

O objetivo do presente estudo foi avaliar a concentração e viabilidade da fração de células mononucleares (FCM) a partir de diferentes técnicas de colheita e processamento de medula óssea (MO) em equinos. Foram avaliados cinco equinos adultos, hígidos e sem raça definida. Obtiveram-se frações de medula óssea (MO) do osso esterno, de acordo com dois protocolos: na colheita A, utilizou-se 10mL de solução de heparina dentro da seringa e em seguida, aspirou-se a MO; na colheita B, 10mL de solução de heparina foi injetada na MO e a aspiração foi realizada após 20 segundos. Todos os animais foram submetidos aos dois protocolos de colheitas, realizadas em sequência, sem intervalo entre os dois procedimentos. Após isolamento da fração de células mononucleares (FCM), das amostras de MO obtidas nas colheitas A e B, cada amostra foi dividida em dois tubos, um contendo solução de DMEM e outro contendo PBS. Assim, alternando-se o tipo de colheita e a solução diluidora, obteve-se quatro tubos de amostras por animal. Os tubos foram centrifugados e os sedimentos foram homogeneizados nos respectivos meios obtendo-se o volume final de 100μL. Realizou-se determinação da concentração e viabilidade celular, obtendo-se as concentrações médias de FCM. Para ambos os meios de diluição, a colheita B apresentou valor numérico maior em comparação à colheita A, porém não foi significativo (p>0,05). Atribui-se tal tendência à menor ocorrência de coagulação da MO no momento da colheita B, sugerindo-se melhor aproveitamento da FCM. Não houve diferença (p>0,05) entre os meios DMEM ou PBS, indicando que os mesmos não alteraram a viabilidade celular. Os protocolos utilizados para colheita de MO e separação da FCM se mostraram eficientes, para o uso em terapia celular em equinos.

The aim of this study was to evaluate mononuclear cells fraction (MCF) concentration and viability from different techniques of bone marrow (BM) aspiration and processing in horses. Five adult horses, healthy and of unknown breed were evaluated. BM was obtained from sternum bone, according two protocols: in aspiration A, 10mL of heparin solution was used inside the syringe and BM was aspirated; in aspiration B, 10mL of heparin solution was injected into the BM, and aspiration was done after 20 seconds. All the animals were submitted by both protocols realized in sequence, without a gap between the procedures. After MCF isolation, of BM samples obtained from A and B aspiration, each sample was divided into two tubes; one contained DMEM solution and the other with PBS solution. Therefore, interchanging the aspiration protocol and the dilution solution, four sample tubes were obtained for each horse. The tubes were centrifuged and the pellet was homogenized with the respectively solution to obtain the final volume of 100μL. Cellular concentration and viability were determined to obtain the FCM medium concentration. For both solutions, the aspiration B had higher numeric values comparing with aspiration A; however, it was not significant (p>0.05). This tendency is attribute for the less BM coagulation observed in the aspiration B, suggesting greater improvement of MCF. No difference (p>0.05) was found between DMEM and PBS solution, indicating that both do not alter the cell viability. The protocols used for BM aspiration and MCF isolation were efficient for application in equine cellular therapy.

Viabilidad de una bacteria láctica encapsulada e incorporada en una matriz de cobertura de chocolate / Viability of encapsulated lactic bacteria added in a matrix of chocolate coverage

Se evaluó la viabilidad durante el almacenamiento de Weissella confusa incorporada en una matriz de cobertura de chocolate. La bacteria probiótica se encapsuló empleando tres materiales de pared, gel de Aloe vera, gel Aloe vera + Almidón al 10 % y gel de Aloe vera + Almidón al 15 % y células libres como control. Posteriormente se liofilizó. La bacteria probiótica encapsulada, se incorporó en una matriz de cobertura de chocolate. Los chips se empacaron y almacenaron durante 5 semanas a 4 °C, cada semana se midieron cambios en la viabilidad de la bacteria probiótica y en la actividad de agua. En la quinta semana, los chips se sometieron a condiciones simuladas de jugos intestinales. Durante el almacenamiento los chips mantuvieron su carácter probiótico (>10(6) UFC/g), sin embargo, cuando la bacteria probiótica se encapsuló en gel aloe vera, se obtuvo mayor número de bacterias probióticas vivas dentro de la matriz sólida (2,1x10(8) UFC/g). La actividad de agua varió de 0,470 a 0,810. La bacteria probiótica permaneció viva por 2 horas en medios simulados de jugos intestinales, lo cual ratifica que la matriz sólida y los medios de encapsulación seleccionados son adecuados para el desarrollo de productos sólidos probióticos ricos en grasa vegetal.

Viability during storage of Weissella confusa incorporated in a chocolate coating matrix was evaluated. Probiotic bacteria was encapsulated using three wall materials, Aloe vera gel, Aloe vera gel + 10 % starch and aloe vera gel + 15 % starch and free cells as control. Subsequently lyophilized. Probiotic bacteria encapsulated, was incorporated into a chocolate coating matrix. The chips were packed and stored for 5 weeks at 4 °C, were measured weekly changes in viability of the probiotic bacteria and water activity. In the fifth week, the chips were subjected to simulated conditions of intestinal juices. During storage chips remained probiotic character (>10(6) CFU/g), however, if the probiotic bacteria are encapsulated in aloe vera gel, the greater number of living probiotic bacteria was obtained within the solid matrix (2,1x10(8) CFU/g ). Water activity ranged from 0.470-0,810. Probiotic bacteria remained alive for 2 hours in simulated intestinal fluid media, which confirms that the solid matrix and the selected encapsulation means are suitable for the development of solid product rich in vegetable fat probiotics.

Construction and Application of Lentiviral Vectors Expressing microRNA-203 Tough Decoy / 天津医药

Objective To establish method of constructing lentiviral vectors to express microRNA (miRNA) ''tough decoy''(TuD)and to detect the effects of the TuD on cellular endogenous miRNA level and cellular phenotypes. Methods Two-step cloning strategy was utilized to first generate a universal miRNA TuD frame vector,followed by con-structing the TuD expression vector specially targeting miR-203. The package of the recombinant lentivirus was per-formed in 293T cells. Then the rat bone marrow mesenchymal stem cells(BM-MSCs)were infected by the miR-203 TuD expression lentivirus. The pSIH1-H1-copGFP vector was also packaged and the BM-MSCs infected by this lentivirus were served as control. Endogenous miR-203 level in BM-MSCs was measured by quantitative RT-PCR,and cellular vi-ability and apoptosis were detected by CCK-8 test and Annexin V-PI staining respectively. Results The miR-203 TuD expression vector was successfully constructed and inserted sequence was validated. At the 3rd,6th and 9th days after in-fected by the miR-203 TuD expression lentivirus,rat BM-MSCs exhibited a repressed endogenous miR-203 level. The miR-203 TuD also promoted viability and inhibited apoptosis of BM-MSCs. All these differences between miR-203 TuD group and control group were statistically significant. Conclusion The two-step cloning method for the construction of miRNA TuD expression vector is simple and efficient. The miRNA TuD can effectively suppress the level of the target miRNA and affect cellular phenotypes.

Alcoholic fermentation by the wild yeasts under thermal, osmotic and ethanol stress

This study aimed to explore the variability in the metabolism of nine wild yeasts isolated from the sugarcane juice from a distillery in the Brazilian State of Mato Grosso. Cell viability under the stress conditions was evaluated. The yeasts were inoculated in the test tubes containing sugarcane juice adjusted from 12 to 21º Brix, ethanol from 6 to 12% in volume and temperature at 30, 35 and 40ºC. The viability was established by the growth in Petri dishes and visually by the CO2 production in the test tubes. None of the evaluated yeasts showed simultaneous resistance to the three stress conditions. The potential of yeast BB.09 could be emphasized due to its ability to ferment up to12% ethanol at 30°C.

Estudio comparaiivo de los métodos de resazurin y mtt en estudios de citotoxiccidad en líneas celulares tumorales humanas / Estudi comparatiivo de los métodos de esazurina y mtt en estudios de citotoxicidad en líneas celulares tumorales humanas

Se realizó un estudio comparativo de los métodos de resazurina y MTT como indicadores de medida indirecta de sobrevivencia celular, aplicables para la estimación de actividad citotóxica, con el fin de establecer la idoneidad de cada uno de los dos sistemas de detección, a través de la valoración de citotoxicidad sobre tres líneas celulares derivadas de tumores humanos: HEp-2, HT-29 y HeLa, evaluando tres compuestos sintéticos durante un periodo de tratamiento de 48 horas, usando como referencia de actividad Doxorrubicina HCl, un agente antineoplásico. Los resultados obtenidos por los dos métodos revelaron comportamientos semejantes en las condiciones estudiadas en las líneas celulares.

A study was carried comparing the resazurina and MTT methods as indicators of indirect measure of cellular survival applied to valuated cytotoxic activity in order to assess the suitability of each of the two detection systems. The comparison was made through the valuation of cytotoxicity on three cell lines derived from human tumors: HEp-2, HT-29 and HeLa, evaluating three synthetic compounds during a treatment period of 48 hours using as a reference activity Doxorubicin HCl. The results obtained by the two methods showed a similar performance under the conditions tested in cell lines.