Chemical composition, antiproliferative and antioxidant attributes of ethanolic extract of resinous sediment from Etlingera elatior (Jack. ) inflorescence

The ethanolic extract of resinous sediment (EERS) of Etlingera elatior young inflorescence was examined for its anticancer effect and potential antioxidant activity. The anticancer effect of the EERS was evaluated on four human cancer cell lines, HCT 116, HT-29, Hela, and MCF-7, using the MTT assay. GC-MS analysis showed that the main components found in the EERS were nonyl cyclopropane (4.44%), 1-tetradecane (3.66%), cyclotetradecane (2.41%), cyclododecane (1.92%), and 1-decene (1.72%). The antioxidant activity was determined through different methods. High amounts of TPC and TFC in the EERS were found. Moderate antioxidant capacity of the EERS was detected by DPPH and ABTS assays, with EC50 values of 44.19 and 56.61 µg/mL and a high FRAP value of 281.79 nmol Fe+2 equivalent/mg extract. In the MTT assay, the EERS showed potent anticancer activity, with IC50 values of 19.82, 37.001, 50.49, and 53.29 µg/mL against HT-29, HCT 116, Hela, and MCF-7 tumour cell lines, respectively. Moreover, the results were comparable to or less potent than the standard reference drug, 5-fluorouracil. The results showed that the EERS of Etlingera elatior inflorescence contained a high amount of polyphenols and flavonoids, which may to the selective antiproliferative effects towards colon cancer in vitro

Leaf anatomy of Protium ovatum and its antiproliferative potential in cervical cells

ABSTRACT The aim of this study was to analyze the morphology and anatomy of the leaves of Protium ovatum Engl., Burseraceae, and verify the antiproliferative activity in cervical cells. For anatomical analysis, the leaf samples were fixed in formol, acetic acid, alcohol 70, dehydrated, included in hydroxyethyl methacrylate and sectioned at a thickness of 5-10 µm in rotative microtome. The samples were stained with toluidine blue and blades mounted with synthetic resin "Entellan". Histochemical tests and scanning electron microscopy were also performed. To investigate the antiproliferative effect we used the cells strain of human cervix carcinoma and normal keratinocytes. The anatomical analysis demonstrated that the leaf is hypostomatic and the epidermal cells walls were slightly undulate on both faces. The palisade parenchyma occupies most part of leaf mesophyll. The spongy parenchyma is organized into 3-4 layers of cells. Vascular bundles of smaller diameter and secretory cavities are distributed along the leaf mesophyll. The midrib region was formed by a single vascular bundle with xylem in the center surrounded by phloem. Secretory cavities are distributed along the phloem. The histochemical tests revealed the presence of lipids in the secretory cavities and phenolic compounds in almost cell of mesophyll. Scanning electron microscopy analysis showed the smooth leaf cuticle ornamentation with some striated areas. It was observed antiproliferative effect on human cervix carcinoma cell comparing with normal cells.

Toxicity of Jatropha curcas L. latex in Allium cepa test / Toxicidade do látex de Jatropha curcas L. no modelo de Allium cepa

The latex obtained from Jatropha curcas (physic nut) is used in traditional medicine to treat a variety of disturbs, including burns, hemorrhoids, ringworm and ulcers. Phytochemical analyses have shown that J. curcas latex contains natural compounds with therapeutic potential. In this study, the toxicity, cytotoxicity and genotoxicity effects of J. curcas latex on the root cells of Allium cepa were examined. Onion seeds and bulbs were exposed to seven different concentrations of latex and then the roots were submitted to macro and microscopic analyses. Water and sodium azide were used as negative and positive controls, respectively. The analysis of root growth showed that J. curcas crude latex or 50% diluted is highly toxic. Cytogenetic results showed that the mitotic index of the onion roots submitted to latex treatment decreased significantly compared to the negative control, which suggests that the latex is cytotoxic. High incidence of chromosome aberrations in the cells treated with J. curcas latex was observed too, indicating that the latex also presents genotoxic effect. The analyses presented in this report suggest the toxic, cytotoxic and genotoxic effects of J. curcas latex. Then, the indiscriminate use of J. curcas latex in folk medicine could bring risk to human health.

O látex obtido de Jatropha curcas (pinhão manso) é usado na medicina tradicional para tratamento de diversos distúrbios, como queimaduras, hemorroida, micose e úlcera. Análises fitoquímicas apontaram que o látex de J. curcas contém compostos naturais com potencial terapêutico. Este estudo avaliou a toxicidade, citotoxicidade e genotoxicidade do látex de J. curcas em células da raiz de Allium cepa. Sementes e bulbos de cebola foram expostos á sete diferentes concentrações de látex e, então, as raízes foram submetidas a análises macro e microscópica. Água e azida sódica foram utilizadas como controle negativo e positivo, respectivamente. A análise do comprimento das raízes mostrou que o látex de J. curcas puro e diluído a 50% é altamente tóxico. O índice mitótico das raízes de cebola submetidas ao tratamento com o látex diminuiu significativamente comparado com o controle negativo, o que sugere que o látex é citotóxico. Uma alta incidência de aberrações cromossômicas em células tratadas com o látex de J. curcas também foi observada, indicando que o látex apresenta efeito genotóxico. Essa análise sugere que o látex de J. curcas possui efeitos tóxico, citotóxico e genotóxico, sendo que o uso indiscriminado do látex de J. curcas na medicina popular pode trazer risco à saúde humana.

Lupeol and its esters: NMR, powder XRD data and in vitro evaluation of cancer cell growth

ABSTRACT The triterpene lupeol (1) and some of its esters are secondary metabolites produced by species of Celastraceae family, which have being associated with cytotoxic activity. We report herein the isolation of 1, the semi-synthesis of eight lupeol esters and the evaluation of their in vitro activity against nine strains of cancer cells. The reaction of carboxylic acids with 1 and DIC/DMAP was used to obtain lupeol stearate (2), lupeol palmitate (3) lupeol miristate (4), and the new esters lupeol laurate (5), lupeol caprate (6), lupeol caprilate (7), lupeol caproate (8) and lupeol 3',4'-dimethoxybenzoate (9), with high yields. Compounds 1-9 were identified using FT-IR, 1H, 13C-NMR, CHN analysis and XRD data and were tested in vitro for proliferation of human cancer cell activity. In these assays, lupeol was inactive (GI50> 250µg/mL) while lupeol esters 2 -4 and 7 - 9 showed a cytostatic effect. The XRD method was a suitable tool to determine the structure of lupeol and its esters in solid state. Compound 3 showed a selective growth inhibition effect on erythromyeloblastoid leukemia (K-562) cells in a concentration-dependent way. Lupeol esters 4 and 9 showed a selective cytostatic effect with low GI50 values representing promising prototypes for the development of new anticancer drugs.

Metotrexato en condromatosis sinovial: una serendipia / Methotrexate in sinovial chondromatosis: a case of serendipity

La condromatosis sinovial es una metaplasia idiopática benigna de la membrana sinovial que afecta a 1/100.000 habitantes, en una relación hombre/mujer de 3 a 1 entre los 30 y 50 años. Predomina en grandes articulaciones como rodilla (70%), cadera (20%) y hombro (19%), y en menor proporción en codo y tobillo. Puede ser primaria o secundaria. La etiología es desconocida. La resolución es quirúrgica ya sea por artroscopia o por cirugía a cielo abierto, no existiendo otra alternativa terapéutica. Se presenta el caso clínico de un paciente con condromatosis sinovial en hombro derecho, que se comporta como una artropatía erosiva, indicándose metotrexato y resolviendo casi totalmente los nódulos condromatosos.

The synovial chondromatosis is a benign idiopathic metaplasia ofthe synovial membrane which affects one in 100,000 inhabitants. Itis 3 times more common in males, aged between 30 and 50 yearsold. It is commonly found in large joints such as knee (70%), hip(20%) and shoulder (19%) and less frequently in elbow and heel. Itcan be primary or secondary. The etiology is still unknown.The resolution is surgical by means of arthroscopy or open surgery,existing no other therapeutic alternatives.We present a male patient with primary synovial chondromatosis inthe right shoulder, leading to an erosive arthropathy. Treatment withmethotrexate resolved almost entirely the cartilaginous nodules.

Genotoxic and chromatographic analyses of aqueous extracts of Peltodon longipes Kunth ex Benth. (hortelã-do-campo)

Peltodon longipes is used as a stimulant and emmenagogue. The objective of this study was to perform genotoxic and chromatographic analyses of the extracts of two samples of P. longipes, collected from the cities of Santa Maria and Tupanciretã, RS, Brazil. The Allium cepa assay was used to analyze genotoxicity while high-performance liquid chromatography was employed to determine phenolic compounds. The genotoxicity experiment consisted of nine groups each comprising four A. cepa bulbs. Bulb roots were developed in distilled water and then transferred for the treatments, for 24 hours, and the negative control remained in water. The treatments were: aqueous extracts at concentrations of 5 and 15 g L-1 for each sample, plus four groups treated with 1% glyphosate, one of which was used as a positive control and the other three for testing DNA damage recovery using water and the extracts of P. longipes from Santa Maria. All extracts of P. longipes exhibited anti-proliferative potential, although the effect was significantly greater for the extracts from the Tupanciretã sample. This sample also contained the highest amount of rosmarinic acid and kaempferol, which may confer the effects found in these extracts. Only extracts from the Santa Maria sample exhibited genotoxic potential.

Peltodon longipes é utilizada como estimulante e emenagoga. Objetivou-se realizar análises genotóxica e cromatográfica dos extratos de duas amostras de P. longipes, coletadas nos municípios de Santa Maria e Tupanciretã, RS, Brasil. O teste de Allium cepa foi utilizado para análise da genotoxicidade e a cromatografia líquida de alta eficiência, para determinação dos compostos fenólicos. O experimento de genotoxicidade constou de nove grupos de quatro bulbos de A. cepa. Os bulbos foram enraizados em água destilada e após transferidos para os tratamentos, por 24 horas, permanecendo o controle negativo em água. Os tratamentos foram: extratos aquosos nas concentrações de 5 e 15 g L-1 de cada amostra, além de quatro grupos tratados com glifosato 1%, um deles usado como controle positivo e outros três para testar a recuperação de danos ao DNA, utilizando água e os extratos de P. longipes da amostra de Santa Maria. Todos os extratos de P. longipes demonstraram potencial antiproliferativo, porém o efeito foi significativamente maior para os extratos da amostra de Tupanciretã. Essa amostra também apresentou maior quantidade de ácido rosmarínico e canferol, o que pode estar relacionado com os efeitos encontrados nesses extratos. Somente extratos da amostra de Santa Maria demonstraram potencial genotóxico.

Cytotoxicity of aqueous extracts of Rosmarinus officinalis L. (Labiatae) in plant test system / Citotoxicidade de extratos aquosos de Rosmarinus officinalis L. (Labiatae) em sistema-teste vegetal

This study investigated the cytotoxic activity of Rosmarinus officinalis L. (rosemary) aqueous extract on the cell cycle of Allium cepa. To this end, crude aqueous leaf extracts at four concentrations, 0.02, 0.04, 0.06 and 0.08 mg/mL, were tested on A. cepa meristematic root cells, at exposure times of 24 and 48h. Slides were prepared by the crushing technique, and cells analyzed throughout the cell cycle, totaling 5,000 for each control group and concentration. The four concentrations tested, including the lowest and considered ideal for use, at all exposure times, showed a significant antiproliferative effect on the cell cycle of this test system and presented a high number of cells in prophase. Our results evidenced the cytotoxicity of rosemary extracts, under the studied conditions.

Neste estudo investigou-se a ação citotóxica do extrato aquoso de Rosmarinus officinalis L. (alecrim) sobre o ciclo celular de Allium cepa. Para isso obteve-se extratos aquosos brutos de folhas secas desta planta em quatro concentrações, 0,02; 0,04; 0,06 e 0,08mg/mL, que foram testadas em células meristemáticas de raízes de A. cepa, nos tempos de exposição 24 e 48h. As lâminas foram feitas pela técnica de esmagamento, e analisaram-se células em todo ciclo celular, totalizando 5.000 para cada grupo controle e concentração. A partir dos resultados verificou-se que as quatro concentrações testadas, inclusive a menor e considerada ideal para consumo, em todos os tempos de exposição tiveram ação antiproliferativa significativa sobre o ciclo celular deste sistema teste, e apresentaram um grande número de células em prófase. Dessa forma, o alecrim, nas condições analisadas, mostrou-se citotóxico.

El extracto acuoso de ruta graveolens del norte de méxico causa apoptosis y muestra efecto antiproliferativo sobre el hígado de rata wistar: evidencia morfológica / Aqueous extract of ruta graveolens from the north of mexico causes apoptosis and shows antiproliferative effect on the liver of wistar rat: morphological evidence

Ruta graveolens es una planta nativa del Mediterráneo Oriental y del área Sur Occidental de Asia, de esta planta se han aislado más de 120 compuestos químicos. En un estudio previo en nuestro laboratorio se observó que un extracto acuoso de R. graveolens causó necrosis y alteraciones morfológicas sugestivas de apoptosis sobre el hígado de rata Wistar. El objetivo del presente estudio, fue evaluar la inducción de apoptosis y el posible efecto antiproliferativo in vivo de un extracto acuso de R. graveolens del norte de México, mediante métodos inmunohistoquímicos. Se utilizaron 25 ratas Wistar y se dividieron en 5 grupos (n=5). El grupo 1 correspondió al grupo control negativo, el grupo 2 o control positivo se trató con 100 mg de dexametasona/kg/día. Los grupos 3 y 4 se trataron con 30 y 100 mg de extracto de R. graveolens/kg/día respectivamente. Al grupo 5 se le administraron 100 mg de dexametasona/kg/día combinados con 100 mg de extracto de R. graveolens/kg/día. Las administraciones se realizaron vía intraperitoneal por tres días. Los animales se sacrificaron por dislocación cervical, y se tomaron muestras de hígado que se fijaron en formalina, posteriormente se incluyeron en bloques de parafina. Se obtuvieron cortes histológicos que se tiñeron con el método tricrómico de Masson. También se realizaron pruebas inmunohistoquímicas de TUNEL, anti-bcl-2 y anti-PCNA; además de un estudio morfométrico. Los resultados demuestran por primera vez el potencial apoptósico y antiproliferativo del extracto acuoso de R. graveolens del norte de México, sobre el hígado de rata Wistar. Se sugiere la posibilidad de emplear dosis menores a las administradas en este estudio del extracto acuoso de R. graveolens, para investigar su potencial uso como agente antineoplásico en estudios in vitro con líneas celulares tumorales e/o implantadas en modelos murinos de cáncer.

Ruta graveolens, is a native plant of the Eastern Mediterranean and the South Western area of Asia. From this plant, more than 120 chemical compounds have been isolated. In a previous study in our laboratory, we observed that an aqueous extract of R. graveolens, caused necrosis and morphological alterations suggestive of apoptosis on the liver of Wistar rats. The objective of this study, was to evaluate the induction of apoptosis and a possible antiproliferative effect in vivo of an aqueous extract of R. graveolens from the north of Mexico, by immunohistochemical methods. 25 Wistar rats were used and divided into 5 groups (n= 5). Group 1 corresponded to negative control group, group 2 or positive control was treated with 100 mg of dexamethasone/kg/day. Groups 3 and 4 were treated with 30 and 100 mg of extract of R. graveolens/kg/day respectively. Group 5 received the administration of 100 mg of dexamethasone/kg/day combined with 100 mg of extract of R. graveolens/kg/day. The administrations were by intraperitoneal via for three days. The animals were sacrificed by cervical dislocation, liver samples were taken, fixed in formalin and then samples were embedded in paraffin blocks. Histological sections were obtained and stained with Masson trichrome method. Immunohistochemical assays of TUNEL, anti-bcl-2, and anti-PCNA were performed. Also a morphometric study was carried out. Results show for the first time the potential apoptotic and antiproliferative effect of an aqueous extract of R. graveolens from the north of Mexico on the liver of Wistar rats. This suggests the use of lower doses of the extract of R. graveolens, to investigate its potential use as an antineoplastic agent, in studies in vitro with tumor cell lines and/or implanted in murine models of cancer.

In Vitro Evaluation of the Anticancer Effect of Methanolic Extract of Alstonia scholaris Leaves on Mammary Carcinoma.

Breast cancer is the leading cause of cancer associated death among women worldwide. Current cancer treatments include chemotherapy, radiotherapy, and surgery. However, since these conventional methods often have undesirable side effects,new focus towards the use of plant extract to treating cancer with eliminating the side effects. The objective of present study is to asses the anticancer effect of leaves of Alstonia scholaris, using the cytosolic marker enzymes like Aspartate transaminase (AST),Acid phosphatase(ACP) , Alkaline phosphatase(ALP), Lactate dehydrogenase (LDH), Gamma - glutamyl transferase(γ– GT), and 5'-nuclcotidase(5'-NT) in vitro over breast cancer tissue. These are key enzymes in the metabolic pathways and these are the target for the drugs used in chemotherapy. An elevated level of enzyme concentration signals the presence of malignancy .The effect of leaves of Alstonia scholaris is also assessed by studying the effect of non-enzymatic antioxidants like vitamin A, E, C in vitro over breast cancer tissue. The present study revealed the leaves of methanolic extracts of Alstonia scholaris on cancer cells/tumor cells in vitro has been justified by its cytotoxic effect and anti proliferative effect.

Alterations in the mitotic index of Allium cepa induced by infusions of Pluchea sagittalis submitted to three different cultivation systems

We evaluated the antiproliferative effect of infusions from Pluchea sagittalis using the Allium cepa test. Infusions in three concentrations (2.5, 5, and 25 g dm-3) of leaves cultivated in three environments (in vitro, acclimatized growth chamber, and field) were used. Six onion bulbs were used for each of the eight treatments, and the mitotic index was obtained from 6000 cells per treatment. In conclusion, leaf infusions of P. sagittalis cultivated in the field have a high antiproliferative activity, as well as the cultivation system influences the antiproliferative potential.

Avaliou-se o efeito antiproliferativo de infusões de Pluchea sagittalis usando o teste de Allium cepa. Foram usadas infusões em três concentrações (2,5, 5 e 25g dm-3) de folhas cultivadas em três ambientes (in vitro, sala de crescimento climatizada e em campo). Foram usados seis grupos de bulbos para cada um dos 8 tratamentos e o os índices mitóticos foram obtidos a partir de 6000 células por tratamento. Concluiu-se que a infusão de folhas de P. sagittalis cultivadas em campo possui grande atividade antiproliferativa, bem como o sistema de cultivo de plantas influencia o potencial antiproliferativo.

The Mechanisms of the Antiproliferative Effect by Interferon- a in Cervical Cancer Cell Lines / 대한부인종양콜포스코피학회잡지

Interferons(IFNs) exhibit an antiproliferative effect on many normal and transformed cells and have in vivo antitumor activity in a variety of cancers. Recent clinical studies have suggested major activity with IFNs, especially in advanced squamous cell carcinoma of the skin and cervix. With the objective of exploring how the IFNs might work in squamous carcinoma cell line, we studied the effect of IFN-a on cervical cancer cell lines. The effect of IFNs on apoptosis and cell cycle of cervical cancer cell lines(C33A, CaSki, SiHa, HeLa, ME-180) was analysed by flow cytometry in time dependent manner. Results were as follows: (1) According to cell count of studied cancer cell lines treated with 2,000 IU/ml IFN-a for 7 days exposure, IFN-a had the antiproliferative effect on all five tested cervical cancer cell lines. Also this antiproliferative effect was confirmed by WST-1 assay. (2) The effect of IFN-a on apoptosis of each cultute was analysed by flow cytometry after 3 days and 7 days exposure with 2,000 IU/ml IFN-a, Apoptosis was not induced by IFN-a treatment. (3) The effect of IFN-a on the cell cycle of each culture was analysed by flow cytometry after 3 days exposure with 2,000 IU/ml IFN-a. As compared to control cells, treatment with IFN-a resulted in a higher proportion of cells in S phase with lower portion of cells with G2/M phase. (4) Time course of IFN-a effect on HPV 16 and HPV 18 E6 mRNA levels was evaluated by northern blot analysis. In CaSki cell line, HPV 16 E6 mRNA expression induced by IFN-a was not inhibited. But in HeLa cell line, HPV 18 E6 inRNA expression was inhibited. IFN-a appears to have the antiproliferative effect on all five tested cervical cancer cell lines and the antiproliferative effect of IFN-a seemed to be induced not by apoptosis but by disruption on specific cell cycle. Also regulation of HPV E6 mRNA expression induced by IFN-a is not directly related to the mechanisms of the antiproliferative effect of IFN-a in cancer cell lines.