In-vitro Antioxidant Activity and Flow Cytometric Analysis of Simarouba glauca DC Bark Extract Induced Apoptosis in Triple Negative Breast Cancer Cells.

OBJECTIVE: Ethnomedicinally Simarouba glauca DC is an important plant containing major class of phenols and terpenoids as bioactive compounds. The present study focuses on the evaluation of the anticancer effects of S. glauca bark UAE-EA (Ultrasonicator Assisted Extraction - Ethyl Acetate) fraction (SG-Fraction) against MDA-MB-231 triple negative breast cancer cell lines. METHODS: UAE-EA technique was used for the extraction of phytochemicals from S. glauca bark. Fractionation method was carried out to obtain Ethyl acetate fraction and PPS, TPC, and DPPH assays were performed to characterize the extract. MTT assay was then applied to analyse the viability of cells and MMP assay to confirm the initiation of drug induced apoptosis. Apoptotic morphology and quantification were assessed by DAPI and Annexin V/propidium iodide (PI) staining. RESULTS: UAE yielded 53g of crude extract in methanol. 16g Ethyl acetate fraction was obtained from fractionation. Phytoconstituents such as alkaloids, phenols, flavonoids, and triterpenoids were detected. The TPC was 278.65 mg GAE/100ml. The SG-Fraction showed maximum 66.38% RSA at 200 µg/ml and IC50 value was 101.72 µg/ml. MMP confirmed the induction of apoptosis. DAPI showed the reduction of nuclei with bright chromatin condensation, blebbing, nuclear fragmentation and apoptotic bodies. Annexin-V FITC/PI study showed 59.48% apoptosis induction. This fraction showed a similar trend of antioxidant effect as compared to ascorbic acid but, prominently lower cell viability than Camptothecin (P<0.005). In line with higher TPC in the SG-fraction, free radical scavenging activity was increased (r = 0.098**, p=0.002) and cell viability was reduced significantly (r = -0.097*** p<0.01). CONCLUSION: These results indicate that UAE-EA fraction of S. glauca bark inhibits the growth of MDA-MB-231 cells and can be considered for further neo-adjuvant chemotherapy drug research.

Experimental investigation on use of hydrotreated Simarouba oil (green diesel) and hydrogen gas in a dual-fuel CI engine.

Hydrogen additives to Simarouba glauca vegetable oil (SO) are a common method for addressing the difficulties in combustion caused by SO's poor physical qualities. This research intends to examine parameters such as performance, emission, and combustion characteristics when hydrogen is used as a direct/indirect addition in a SO-fuelled compression ignition (CI) engine. Hydrogen was directly introduced along with intake air until the knocking limit. Experiment was conducted at different load conduction with SO. Through the hydrotreatment process, hydrogen was used in a roundabout way to convert SO to green diesel. Hydrotreated Simarouba vegetable oil (HTSO) enhanced brake thermal efficiency by 23% at full load, whereas direct hydrogen induction improved it by 9% at knock limit. Hydrogen induction resulted in higher NOx emissions than HTSO, at the expense of a slight rise in smoke emissions. Addition of hydrogen lowered both HC and CO emissions directly and indirectly.

Comparison of Simarouba glauca seed shell carbons for enhanced direct red 12B dye adsorption: Adsorption isotherm and kinetic studies.

This study aims at the transformation of the waste lignocellulosic biomass, Simarouba glauca seed shell obtained from biofuel industries, into a value-added adsorbent for the removal of dyes from aqueous media. The basic dye direct red (DR) 12B was adsorbed using chemically (ZnCl2) and thermally activated Simarouba seed shell carbon (ZASRC and SRC, respectively). Dye removal in batch mode was studied by evaluating adsorbent dosage, contact time, pH, adsorption isotherm and kinetics. Enhanced adsorption of DR12B was attained within 80 min at pH 5 with the maximal adsorption capacity (Q0) of 17.48 and 64.94 mg g-1, for SRC and ZASRC, respectively. Further, the dye removal followed Freundlich isotherm model and pseudo second-order kinetics. The mean-free energy of adsorption demonstrated that dye adsorption onto ZASRC occurs through ion-exchange. Thus, ZASRC can be safely and easily applied for the removal of direct red 12B from aqueous solutions.

Caspase-Dependent Apoptosis Induced by Simarouba Glauca on Human Non-Small-Cell Lung Cancer, A549 Cells.

BACKGROUND: The leaves of Simarouba glauca (S. glauca) have been used as a potential source of anticancer agents in traditional medicine. Attempts have been made to isolate anticancer agents from the leaves of S. glauca. The objective of the present study was to demonstrate the anticancer and apoptotic effect of the leaf extract of petroleum ether (LPE) on human non-small-cell lung cancer A549 cells. METHODS: MTT assay was used to investigate the effect of LPE on the viability of A-549 cells. The apoptotic effect of human lung cancer cells was evaluated using fluorescence staining, acridine orange/ ethidium bromide staining, Hoechst staining, flow cytometry analysis, annexin V staining, and caspase assay. RESULTS: The results showed a direct correlation between the dose and the rate of cytotoxicity. Fluorescence staining revealed apoptotic features, such as blebbing and chromatin condensation. Flow cytometry analysis and annexin V staining revealed phosphatidyl serine externalization. Caspase assay confirmed that the extract inhibited cell death. Caspase 3 expressions indicated that the cell death occurred either through the mitochondrial pathway or the death receptor. CONCLUSIONS: The study revealed that the LPE induced the apoptosis of human non-small-cell lung cancer, A549 cells, either through mitochondrial or death receptor pathway.

The Apoptotic Properties of Leaf Extracts of Simarouba glauca against Human Leukemic Cancer Cells.

BACKGROUND AND OBJECTIVE: Simarouba glauca is a plant belonging to the family of Simaroubaceae. It is a potent source of secondary metabolites. The aim of this study was to evaluate the apoptotic properties of leaf extracts of Simarouba glauca against human leukemic cancer cells. MATERIALS AND METHODS: Cytotoxicity of Simarouba glauca was assessed in the leaf extract of petroleum ether against leukemic cells by MTT assay. To detect the apoptotic features, fluorescence microscopy analysis was done with dual acridine orange/ethidium bromide fluorescent staining and Hoechst staining. To determine the externalization of phosphatidylserine, annexin v staining was done. Mitochondrial or death receptor activation was confirmed by caspase 3 analysis by flow cytometry. RESULTS: This study revealed that Simarouba glauca was able to treat leukemia. Among the four extracts, petroleum ether extract showed a higher order of in vitro anticancer activity. The petroleum ether extract strongly inhibited the proliferation of K562 cell lines with IC50 values of 186 µg/ml. Dual acridine orange/ethidium bromide fluorescent staining and Hoechst staining revealed the characteristic features of apoptosis. Annexin V confirmed early and late stage apoptosis. Caspase-3 analysis revealed that cell death was due to mitochondrial or death receptor activation in mitochondrial pathway. CONCLUSION: These findings suggested that Simarouba glauca leaf extracts inhibited leukemic cells in a time- and dose-dependent manner either through mitochondrial or death receptor activation. The leaf extracts of Simarouba glauca was found to be nontoxic to lymphocytes. It can be concluded that Simarouba glauca is an important source of phytochemicals posing efficacy against leukemic cancer cells.
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Molecular mechanism of Escherichia coli H10407 induced diarrhoea and its control through immunomodulatory action of bioactives from Simarouba amara (Aubl.).

Enterotoxigenic Escherichia coli (ETEC) infection is a major cause of death in children under the age of five in developing countries. ETEC (O78:H11:CFA/I:LT+:ST+) mechanism has been studied in detail with either heat labile (LT) or heat stable (ST) toxins using in vitro and in vivo models. However, there is no adequate information on ETEC pathogenesis producing both the toxins (LT, ST) in BALB/c mice model. In this study, female mice have been employed to understand ETEC H10407 infection induced changes in physiology, biochemical and immunological patterns up to seven days post-infection and the antidiarrhoeal effect of Simarouba amara (Aubl.) bark aqueous extract (SAAE) has also been looked into. The results indicate that BALB/c is sensitive to ETEC infection resulting in altered jejunum and ileum histomorphology. Withal, ETEC influenced cAMP, PGE2, and NO production resulting in fluid accumulation with varied Na+, K+, Cl-, and Ca2+ levels. Meanwhile, ETEC subverted expression of IL-1ß, intestine alkaline phosphatase (IAP), and myeloperoxidase (MPO) in jejunum and ileum. Our data also indicate the severity of pathogenesis reduction which might be due to attainment of equilibrium after reaching optimum rate of infection. Nevertheless, degree of pathogenesis was highly significant (p < 0.01) in all the studied parameters. Besides that, SAAE was successful in reducing the infectious diarrhoea by inhibiting ETEC H10407 in intestine (jejunum and ileum), and shedding in feces. SAAE decreased cAMP, PGE2, and fluid accumulation effectively and boosted the functional activity of immune system in jejunum and ileum IAP, MPO, IL-1ß, and nitric oxide.

Structure of intact chitinase with hevein domain from the plant Simarouba glauca, known for its traditional anti-inflammatory efficacy.

Chitinase from the leaves of Simarouba glauca, a plant used in traditional anti-inflammatory therapy is purified and characterized. Peptide mass finger print analysis revealed the protein as an endo-chitinase which was further confirmed using chitin-agar assay. The enzyme exhibited significant anti-fungal efficacy against phyto-pathogens such as Macrophomina phaseolina, Fusarium oxysporum and Sclerotium rolfsii. Chitinolysis was also examined against insoluble chitin using SEM. Using X-ray diffraction data up to 1.66 Å, the structure was determined by Molecular Replacement using crystal structure of GH19 Chitinase-like protein from Hevea brasiliensis. During structure refinement, an extra domain could be traced and identified as hevein domain. To our knowledge, this is the first report of any chitinase with intact hevein domain. The GH19 chitinase and hevein domains though connected by a lengthy loop, are restricted to be close by disulfide bridges. These bridges connecting each domain with the loop may be important for proper chitin feeding into the active site. By considering reports on hevein and chitinase domains as well as the traditional use of the plant, this report of an intact hevein-chitinase protein and their relative orientation may add further insights for the usefulness of this protein.

Antibacterial Activity of Leaf Extract of Annona muricata and Simarouba glauca on Enterococcus faecalis.

AIM: To determine the antimicrobial effect of water extracts of leaves of Annona muricata and Simarouba glauca on Enterococcus faecalis using agar diffusion method. MATERIALS AND METHODS: Dried leaves of A. muricata and S. glauca were powdered and extracted in a soxhlet apparatus. Enterococcus faecalis was grown overnight in Trypticase soy agar plates. About 10 µl of each extract was placed on agar plates and incubated overnight. The zone of inhibition was measured after 24 hours. About 1% sodium hypochlorite and distilled water were used as positive and negative controls. RESULTS: The leaf extract of A. muricata showed similar effectiveness as that of sodium hypochlorite, whereas the leaf extract of S. glauca showed only a slight reduction in growth of E. faecalis. CONCLUSION: Leaf extract of A. muricata can be developed as an alternative to sodium hypochlorite for root canal irrigants. CLINICAL SIGNIFICANCE: Success of endodontic treatment depends on complete disinfection of the root canals. Root canal irrigants have a major role in complete disinfection of the root canals. Chemical root canal irrigants are more or less toxic to the oral environment. In this study, naturally derived leaf extracts of A. muricata and S. glauca are compared with sodium hypochlorite for its effectiveness against E. faecalis - the most common pathogen found in the root canals.

Evidence of the late lignification of the G-layer in Simarouba tension wood, to assist understanding how non-G-layer species produce tensile stress.

To recover verticality after disturbance, angiosperm trees produce 'tension wood' allowing them to bend actively. The driving force of the tension has been shown to take place in the G-layer, a specific unlignified layer of the cell wall observed in most temperate species. However, in tropical rain forests, the G-layer is often absent and the mechanism generating the forces to reorient trees remains unclear. A study was carried out on tilted seedlings, saplings and adult Simarouba amara Aubl. trees-a species known to not produce a G-layer. Microscopic observations were done on sections of normal and tension wood after staining or observed under UV light to assess the presence/absence of lignin. We showed that S. amara produces a cell-wall layer with all of the characteristics typical of G-layers, but that this G-layer can be observed only as a temporary stage of the cell-wall development because it is masked by a late lignification. Being thin and lignified, tension wood fibres cannot be distinguished from normal wood fibres in the mature wood of adult trees. These observations indicate that the mechanism generating the high tensile stress in tension wood is likely to be the same as that in species with a typical G-layer and also in species where the G-layer cannot be observed in mature cells.

Green technology effect of injection pressure, timing and compression ratio in constant pressure heat addition cycle by an eco-friendly material.

Present energy strategies focus on environmental issues, especially environmental pollution prevention and control by eco-friendly green technologies. This includes, increase in the energy supplies, encouraging cleaner and more efficient energy management, addressing air pollution, greenhouse effect, global warming, and climate change. Biofuels provide the panorama of new fiscal opportunities for people in rural area for meeting their need and also the demand of the local market. Biofuels concern protection of the environment and job creation. Renewable energy sources are self-reliance resources, have the potential in energy management with less emissions of air pollutants. Biofuels are expected to reduce dependability on imported crude oil with connected economic susceptibility, reduce greenhouse gases, other pollutants and invigorate the economy by increasing demand and prices for agricultural products. The use of neat paradise tree oil and induction of eco-friendly material Hydrogen through inlet manifold in a constant pressure heat addition cycle engine (diesel engine) with optimized engine operating parameters such as injection timing, injection pressure and compression ratio. The results shows the heat utilization efficiency for neat vegetable oil is 29% and neat oil with 15% Hydrogen as 33%. The exhaust gas temperature (EGT) for 15% of H2 share as 450°C at full load and the heat release of 80J/deg. crank angle for 15% Hydrogen energy share.

Hepatoprotective effect of the aqueous extract of Simarouba amara Aublet (Simaroubaceae) stem bark against carbon tetrachloride (CCl4)-induced hepatic damage in rats.

Simarouba amara stem bark decoction has been traditionally used in Brazil to treat malaria, inflammation, fever, abdominal pain, diarrhea, wounds and as a tonic. In this study, we investigate the hepatoprotective effects of the aqueous extract of S. amara stem bark (SAAE) on CCl4-induced hepatic damage in rats. SAAE was evaluated by high performance liquid chromatography. The animals were divided into six groups (n = 6/group). Groups I (vehicle-corn oil), II (control-CCl4), III, IV, V and VI were pretreated during 10 consecutive days, once a day p.o, with Legalon® 50 mg/kg b.w, SAAE at doses 100, 250 and 500 mg/kg b.w, respectively. The hepatotoxicity was induced on 11th day with 2 mL/kg of 20% CCl4 solution. 24 h after injury, the blood samples were collected and their livers were removed to biochemical and immunohistochemical analyzes. The SAAE decreased the levels of liver markers and lipid peroxidation in all doses and increased the catalase levels at doses 250 and 500 mg/kg. Immunohistochemical results suggested hepatocyte proliferation in all doses. These results may be related to catechins present in SAAE. Thus, SAAE prevented the oxidative damage at the same time that increased regenerative and reparative capacities of the liver.

Spontaneous and experimental intoxication of cattle by Simarouba versicolor A. St.-Hill (Simaroubaceae).

This study describes an outbreak of Simarouba versicolor intoxication in cattle from Mato Grosso do Sul, Brazil, and reproduces it experimentally. Clinical signs of the affected animals were weakness, tremors, hind limbs incoordination, reluctance to move, sternal and lateral recumbency and death. The main necropsy findings, observed in the abomasum and in segments of the small and large intestines, were diffuse redness and mucosal and serosal swelling. Histological examination revealed necrosis of lymphoid tissues and necrotizing enterocolitis. One experiment was carried out using 3 male calves to test the toxicity of a single dose of S. versicolor leaves at 15 g/kg, 5 g/kg and 2.5 g/kg. Clinical signs, necropsy findings and histological examination of calves receiving 15 g/kg and 5 g/kg leaves were similar to those of cattle from the intoxication outbreak. The calf fed 2.5 g/kg leaves developed clinical symptoms of poisoning and recovered naturally. In a second experiment, two male calves received daily administration of S. versicolor leaves at 1.5 g/kg and 2.5 g/kg for 10 days. They developed clinical signs of intoxication within 24 h and recovered eight to nine days after the leaves were administered. These findings suggest that S. versicolor was responsible for the outbreak studied, although this plant does not have cumulative intoxication effects on cattle.

Características citomorfológicas y microquímicas de las especies medicinales conocidas popularmente como aceituno, siete negritos, jacaranda y hierba del cáncer / Cytomorphological and microchemical characteristics of medicinal species popularly known as aceituno, siete negritos, jacaranda and hierba del cáncer

La flora guatemalteca es un potencial recurso para la investigación de plantas medicinales, la cuales han sido utilizadas como alternativas para tratar enfermedades. En la Universidad de San Carlos de Guatemala se han realizado varios estudios que han comprobado la efectividad terapéutica de las plantas, ya que al ser confirmadas como tales, pueden ayudar a combatir enfermedades infecciosas las cuales representan en nuestro país una de las principales causas de morbilidad y mortalidad. En muchos establecimientos se dispensan plantas que dicen ser medicinales; sin embargo, su identidad puede ser dudosa, debido a que muchos de ellos se confían apenas de las propiedades organolépticas para asegurar la naturaleza de una planta determinada. La adulteración o falsificación de plantas medicinales, puede ocasionar una situación perjudicial en la enfermedad de un paciente, retardando o empeorando su curación, además de la pérdida de confianza en la actividad terapéutica de las plantas. Para lograr la calidad de un producto fitoterápeutico, es necesario describir parámetros que sirvan para verificar la identidad y calidad de una droga vegetal; por esta razón se realizó esta investigación, en la cual se establecieron las características citomorfológicas y microquímicas de cuatro plantas utilizadas medicinalmente: Jacaranda mimosifolia D. Don (jacaranda), Acalypha guatemalensis Pax & Hoffm (hierba del cáncer), Simarouba glauca DC. (aceituno), Lantana camara L. (siete negritos). Para cada una de estas especies se establecieron parámetros de identidad macroscópicas en los cuales se determinó la identidad botánica a través de la observación de la planta y su correcta identificación a través de la elaboración de ejemplares de herbario. Los resultados mostraron que las platas estudiadas presentan las siguientes características: En S. glauca se observó que las características más predominantes son: cutícula muy evidente en la epidermis adaxial, estomas anomocíticos y ciclocíticos, fibras esclerénquimas, abundantes astroesclereidas en todo el mesófilo, parénquima esponjoso extenso con grandes espacios intercelulares, parénquima de almacenamiento de taninos y almidón, pecíolulo bilobulado redondeado con un único haz vascular concéntrico. Lantana camara: hoja de anfiestomáticas, mesófilo dorsiventral e inferior papiloso, epidermis unietratificada compacta, están cubiertas con una cutícula con células de tamaño regular y varios tipos de tricomas glandulares y atenuados en toda su extensión, con estomas de tipo anomocíticos y diacíticos, el sistema vascular es abierto, formando un arco aplanado en la "V" con dos haces accesorios situados dorsalmente venación es de tipo pinnada. En J. mimosifolia no se observó esclerénquima en el tejido de sostén, la forma característica del pecíolo es de forma bilobada con bordes redondeados rústicos con un haz vascular en cada lóbulo, cristales de oxalato de calcio solitarios, se observaron estomas paracíticos, tricomas glandulares y atenuados. A. guatemalensis se observaron abundantes cristales en forma de drusas de oxalato de calcio en toda la hoja, parénquima esponjoso compacto, abundantes diferentes tipos de tricomas en toda la hoja, se evidenciaron células grandes y de diferentes formas en las dos epidermis del mesófilo, con estomas paracíticos. Respecto a los parámetros de pureza, se verificó el porcentaje de humedad del material vegetal resultando inferior al 10% tal y como lo establece la Organización Mundial de la Salud; el porcentaje de cenizas totales se obtuvo mediante tres repeticiones, variando el resultado dependiendo de la especie analizada. Los parámetros anteriormente realizados a cada una de las especies, contribuirán a la elaboración de monografías de control de calidad de las especies nativas en estudio; con la finalidad de garantizar que las drogas vegetales frescas o secas provenientes de estas plantas que se utilicen ya sea en el campo de investigación o en la elaboración de fitofármacos; estén plenamente identificadas antes de su uso y/o comercial.

Effectiveness of seed dispersal by ants in a Neotropical tree.

Seed dispersal and subsequent recruitment is the template on which forest regeneration takes place. Hence, considering the scale over which ecological processes occur is key for understanding the overall impact of various dispersal agents. To explore leafcutter ant (Atta colombica) dispersal effectiveness in space and time, seed movement and subsequent recruitment of a large-seeded predominately vertebrate-dispersed tree, Simarouba amara (Aubl. Simaroubaceae), was investigated on Barro Colorado Island, Panama. At each of 218 reproductive-sized adults (≥20 cm diameter at breast height), presence or absence of a leafcutter ant colony was noted, with extensive checks for Atta activity taking place at or in close proximity to seed and seedling transects, which extended 4 cardinal directions for 30 m from each reproductive female tree (n= 74). Only at 2 S. amara trees were nests observed, and in these areas a dense S. amara seedling carpet was observed. Although nearby nest and dump sites might increase local S. amara recruitment in the short term, mortality at these sites is complete or nearly so. Hence, the seed dispersal effectiveness by leafcutter ants appears to be ephemeral and likely contributes inconsequentially to the long-term recruitment and distribution patterns of the species. This finding highlights the importance of evaluating disperser effectiveness at ecologically relevant spatiotemporal scales.

Induction of murine embryonic stem cell differentiation by medicinal plant extracts.

Epidemiological evidence indicates that diets high in fruits and vegetables provide a measure of cancer chemoprevention due to phytochemical constituents. Natural products are a rich source of cancer chemotherapy drugs, and primarily target rapidly cycling tumor cells. Increasing evidence indicates that many cancers contain small populations of resistant, stem-like cells that have the capacity to regenerate tumors following chemotherapy and radiation, and have been linked to the initiation of metastases. Our goal is to discover natural product-based clinical or dietary interventions that selectively target cancer stem cells, inducing differentiation. We adapted an alkaline phosphatase (AP) stain to assay plant extracts for the capacity to induce differentiation in embryonic stem (ES) cells. AP is a characteristic marker of undifferentiated ES cells, and this represents a novel approach to screening medicinal plant extracts. Following a survey of approximately 100 fractions obtained from 12 species of ethnomedically utilized plants, we found fractions from 3 species that induced differentiation, decreasing AP and transcript levels of pluripotency markers (Nanog, Oct-4, Rex-1). These fractions affected proliferation of murine ES, and human embryonal, prostate, and breast carcinoma cells in a dose-dependent manner. Several phytochemical constituents were isolated; the antioxidant phytochemicals ellagic acid and gallic acid were shown to affect viability of cultured breast carcinoma cells.

Estudo do crescimento e qualidade de mudas de marupá (Simarouba amara Aubl. ) em viveiro / Study of growth and quality of marupá (Simarouba amara Aubl. ) nursery seedlings

O objetivo do trabalho foi avaliar o crescimento e a qualidade das mudas de marupá com o sistema radicular podado em diferentes sombreamentos. O experimento foi instalado na Estação Experimental de Silvicultura Tropical do INPA, em Manaus (AM), situada na rodovia BR-174, km 43. As sementes foram beneficiadas e semeadas a 1 cm de profundidade em areia lavada. As mudas foram repicadas para sacos plásticos contendo 3 partes de terra de subsolo, 1 de areia, 0,5 de esterco de galinha curtido. Após as podas das raízes em 0 por cento, 25 por cento, 50 por cento e 75 por cento as mudas permaneceram por 30 dias sob galpão, irrigadas diariamente e depois levadas para canteiros cobertos com telas sombrite de 30 por cento, 50 por cento, 70 por cento e 0 por cento (sem sobreamento). Cada parcela continha 35 mudas incluindo bordadura simples. Foram avaliadas 5 mudas (repetições) ao acaso aos 57, 139 e 182 dias em viveiro. Os dados foram analisados por fatorial 4 x 4 na primeira e 3 x 4 nas outras medições. Foi estudada a altura total (HT), diâmetro do colo (DC), número de folhas, área foliar, pesos da matéria seca de raiz, caule e folhas e a qualidade das mudas pelo Índice de Qualidade de Dickson e pela relação HT/DC. As mudas foram atacadas por lagartas nos canteiros sem sombreamento e a sobrevivência foi superior a 83 por cento naquelas sombreadas. As mudas resistiram às podas e, aos 182 dias, apresentaram maior qualidade para plantios e com maior equilíbrio de crescimento sob sombreamento de 50 por cento.

Our objective was to evaluate the growth and quality of marupá (Simarouba amara Aubl.) seedlings with root system pruning in different shadings. The experiment was undertaken at the INPA Tropical Forestry Experimental Station, in Manaus (AM), located at highway BR-174, km 43. The seeds were processed and sown at a depth of 1 cm in washed sand. The seedlings were transplanted to plastic bags containing 3 parts subsoil, 1 sand, and 0.5 chicken manure fertilizer. After pruning their roots at 0 percent, 25 percent, 50 percent and 75 percent, for 30 days the seedlings remained under the shed, irrigated daily and then planted in beds covered with screens of 30 percent, 50 percent, 70 percent and 0 percent ( without shading). Each plot contained 35 seedlings including simple edge. Five seedlings (repetitions) were assessed at random at 57, 139 and 182 days in nurseries. The data were analyzed by factorial 4 x 4 in the first and 3 x 4 in the other measurements. We studied the total height (HT), diameter of the colon (DC), number of leaves, leaf area, dry weight of roots, stems and leaves of the seedlings, the quality index of Dickson, and the relationship HT/DC. The seedlings were attacked by caterpillars in beds without shading and survival was over 83 percent in shaded beds. The seedlings survived pruning, and at 182 days, showed higher quality for plantations, and better growth balanceunder 50 percent shade.

Cytotoxic activity of Brazilian Cerrado plants used in traditional medicine against cancer cell lines.

UNLABELLED: The search for new anti-cancer drugs is one of the most prominent research areas of natural products. Numerous active compounds isolated from Brazilian Cerrado plant species have been studied with promising results. AIM OF THE STUDY: To investigate the cytotoxic potential of 412 extracts from Brazilian Cerrado plants used in traditional medicine belonging to 21 families against tumor cell lines in culture. MATERIAL AND METHOD: Maceration of 50 plant species resulted in 412 hexane, dichloromethane, ethanol and hydroalcohol extracts. The cytotoxicity of the extracts was tested against human colon carcinoma (HCT-8), melanoma (MDA-MB-435), and brain (SF-295) tumor cell lines, using the thiazolyl blue test (MTT) assay. Bioassay-guided fractionation was performed for one active extract. RESULTS AND CONCLUSIONS: Twenty-eight of the 412 tested extracts demonstrated a substantial antiproliferative effect, at least 85% inhibition of cell proliferation at 50 microg/mL against one or more cell lines. Those extracts are obtained from different parts of Anacardiaceae, Annonaceae, Apocynaceae, Clusiaceae, Flacourtiaceae, Sapindaceae, Sapotaceae, Simaroubaceae and Zingiberaceae. Complete dose-response curves were generated and IC(50) values were calculated for these active extracts against four cell lines HCT-8, MDA-MB-435, SF-295 and HL-60 (leukemia), and their direct cytotoxic effects were determined. In summary, 14 extracts of 13 species showed toxicity in all tested tumor cell lines, with IC(50) values ranging from 0.1 to 19.1 microg/mL. The strongest cytotoxic activity was found for the hexane extract of Casearia sylvestris var. lingua stem bark, with an IC(50) of 0.1 microg/mL for HCT-8, 0.9 microg/mL for SF-295, 1.2 microg/mL for MDA-MB-435, and 1.3 microg/mL for HL-60, and Simarouba versicolor root bark, with an IC(50) of 0.5 microg/mL for HCT-8, 0.7 microg/mL for SF-295, 1.5 microg/mL for MDA-MB-435, 1.1 microg/mL for HL-60. Bioassay-guided fractionation of the last extract led to the isolation of glaucarubinone, which showed pronounced activity against the four cell lines studied. Further studies of the active extracts are necessary for chemical characterization of the active compounds and more extensive biological evaluations.

Studies on protein characteristics and toxic constituents of Simarouba glauca oilseed meal.

In order to exploit the protein rich (47.7 g/100g) simarouba meal in food/feed, studies were conducted on its chemical composition with emphasis on protein characteristics and toxic constituents. Simarouba meal contained high calcium (143 mg/100g) and sodium (79 mg/100g). Saponins with triterpenoid aglycone (3.7 g/100g), alkaloids (1.01 g/100g), phenolics (0.95 g/100g) and phytic acid (0.73 g/100g) were the major toxic constituents identified in simarouba meal. TLC and HPLC results indicated that among different fractions of simarouba saponins, one dominant fraction accounted for about 28%. Proteins of simarouba recorded high in vitro digestibility (88%). SDS-PAGE revealed four major protein bands in molecular weight ranges of 20-24, 36-45 and 55-66 kDa. Apart from, glutamic acid (23.43 g/100g protein) and arginine (10.75 g/100g protein), simarouba protein contained high essential amino acids like leucine (7.76 g/100g protein), lysine (5.62 g/100g protein) and valine (6.12 g/100g protein). Among nutritional indices, simarouba meal recorded a good EAA Index (75.02), C-PER (1.90) and PDCAAS (1.0-Adult group).

In vitro anti-microbial activity of the Cuban medicinal plants Simarouba glauca DC, Melaleuca leucadendron L and Artemisia absinthium L.

In the present study, an extensive in vitro antimicrobial profiling was performed for three medicinal plants grown in Cuba, namely Simarouba glauca, Melaleuca leucadendron and Artemisia absinthium. Ethanol extracts were tested for their antiprotozoal potential against Trypanosoma b. brucei, Trypanosoma cruzi, Leishmania infantum and Plasmodium falciparum. Antifungal activities were evaluated against Microsporum canis and Candida albicans whereas Escherichia coli and Staphylococcus aureus were used as test organisms for antibacterial activity. Cytotoxicity was assessed against human MRC-5 cells. Only M. leucadendron extract showed selective activity against microorganisms tested. Although S. glauca exhibited strong activity against all protozoa, it must be considered non-specific. The value of integrated evaluation of extracts with particular reference to selectivity is discussed.

In vitro anti-microbial activity of the Cuban medicinal plants Simarouba glauca DC, Melaleuca leucadendron L and Artemisia absinthium L

In the present study, an extensive in vitro antimicrobial profiling was performed for three medicinal plants grown in Cuba, namely Simarouba glauca, Melaleuca leucadendron and Artemisia absinthium. Ethanol extracts were tested for their antiprotozoal potential against Trypanosoma b. brucei, Trypanosoma cruzi, Leishmania infantum and Plasmodium falciparum. Antifungal activities were evaluated against Microsporum canis and Candida albicans whereas Escherichia coli and Staphylococcus aureus were used as test organisms for antibacterial activity. Cytotoxicity was assessed against human MRC-5 cells. Only M. leucadendron extract showed selective activity against microorganisms tested. Although S. glauca exhibited strong activity against all protozoa, it must be considered non-specific. The value of integrated evaluation of extracts with particular reference to selectivity is discussed.