Screening of Venezuelan medicinal plant extracts for cytostatic and cytotoxic activity against tumor cell lines.

There are estimated to be more than 20,000 species of plants in Venezuela, of which more than 1500 are used for medicinal purposes by indigenous and local communities. Only a relatively small proportion of these have been evaluated in terms of their potential as antitumor agents. In this study, we screened 308 extracts from 102 species for cytostatic and cytotoxic activity against a panel of six tumor cell lines using a 24-h sulphorhodamine B assay. Extracts from Clavija lancifolia, Hamelia patens, Piper san-vicentense, Physalis cordata, Jacaranda copaia, Heliotropium indicum, and Annona squamosa were the most cytotoxic, whereas other extracts from Calotropis gigantea, Hyptis dilatata, Chromolaena odorata, Siparuna guianensis, Jacaranda obtusifolia, Tapirira guianensis, Xylopia aromatica, Protium heptaphyllum, and Piper arboreum showed the greatest cytostatic activity. These results confirm previous reports on the cytotoxic activities of the above-mentioned plants as well as prompting further studies on others such as C. lancifolia and H. dilatata that have not been so extensively studied.

Effective radiotherapy of primary tumors and metastasis with 18F-2-deoxy-2-fluoro-D-glucose in C57BL/6 mice.

INTRODUCTION: The radiochemical 18F-2-deoxy-2-fluoro-D-glucose (18F-FDG), a positron emitter, is taken up preferentially by malignant tumors with high metabolic rates. This concentration of 18F-FDG in the tumor permits diagnosis and staging by positron emission tomography but also may represent a means of targeting radiotherapy. In this study, we determined the effect of a higher dose of 18F-FDG on tumor growth in a mouse model. MATERIALS AND METHODS: The effect of 18F-FDG on the growth and viability of 3 tumor cell lines was determined in vitro. Primary tumor growth and metastasis of B16/BL6 melanoma cells were determined in C57BL/6 mice injected with 5 mCi doses of 18F-FDG (2/3 doses). RESULTS: 18F-FDG was cytostatic for all 3 cell lines at the lowest dose tested. It significantly reduced the growth of primary tumors, by 89% at day 19 postinoculation, and also almost totally inhibited the appearance of lung metastases after intravenous inoculation of the same cells. CONCLUSIONS: 18F-FDG proved to be an effective radiotherapeutic agent in this model. The possible problems associated with the accumulation of this radiochemical at other sites besides the tumor must be addressed.

Expression of human soluble complement receptor 1 by a pig endothelial cell line inhibits lysis by human serum.

The importance of complement activation and naturally occurring anti-pig antibodies in the hyperacute rejection (HAR) observed in models of pig-to-human xenotransplantation is well established. To overcome this, much effort has been dedicated to preparing transgenic pigs by knocking out Galalpha(1-3)Gal expression in these animals, or knocking in the expression of human complement regulatory proteins (CRPs), such as CD59 or decay accelerating factor. A soluble form of another membrane CRP, complement receptor type 1 (CR1), has also been shown to inhibit complement activation. Here, we show that transfection of a pig endothelial cell line with a truncated form of human soluble complement receptor 1 (sCR1) almost completely protected these cells from complement-mediated lysis by human AB serum. Pigs genetically manipulated to express human sCR1 may represent an additional strategy to inhibit HAR of pig-to-human transplanted organs.

Synthesis and characterization of new palladium-clotrimazole and palladium-chloroquine complexes showing cytotoxicity for tumor cell lines in vitro.

New palladium complexes of chloroquine (CQ) and clotrimazole (CTZ) have been prepared, characterized, and evaluated against four tumor cell lines in vitro. [Pd (CQ)2Cl2] (1) was synthesized by the reaction of PdCl2(CH3CN)2 with CQ, and the [Pd (CTZ)2Cl2] (2) complex by a similar reaction. The new compounds were characterized by a combination of FAB-MS (fast atom bombardment-mass spectrum), elemental analysis, molar conductivity, IR, and NMR spectroscopy. The solid-state structure of 2 has been determined by X-ray crystallography. 2 crystallizes in the monoclinic space group P(2(1)/c), with a = 21.100(4) A, b = 13.408(3) A, c = 22.642(5) A. The structure refinement converged at R1 = 0.0728, wR2 = 0.1918. The cytotoxicity of these two complexes for the tumor cell lines, PANC-1, SKBR-3, MDA-MB231 and HT-29, was compared with that of the original ligands. Ligation of palladium to CTZ led to an increase in the IC50, although a three-fold reduction in the IC50 of CQ was observed on ligation to the metal when tested against the MDA-MB231 cell line.

Ajoene inhibits both primary tumor growth and metastasis of B16/BL6 melanoma cells in C57BL/6 mice.

Ajoene is an organosulphur compound derived from garlic with important effects on several membrane-associated processes such as platelet aggregation, as well as being cytotoxic for tumor cell lines in vitro. In the present study, we investigated the effect of ajoene on different cell types in vitro, as well as its inhibitory effects on both primary tumors and metastasis in a mouse model. We found ajoene to inhibit tumor cell growth in vitro, but also to inhibit strongly metastasis to lung in the B16/BL6 melanoma tumor model in C57BL/6 mice. As far as we are aware, this is the first report of the anti-metastatic effect of ajoene. Ajoene also inhibited tumor-endothelial cell adhesion, as well as the in vivo TNF-alpha response to lipopolysaccharide. Possible mechanisms of its antitumoral activity are discussed in the light of these results.

Effect of 3 plant extracts on b16-bl6 melanoma cell growth and metastasis in C57bl/6 mice

En el presente estudio, se investigaron los posibles efectos de extractos acuosos de tres plantas amazónicas, Uncaria tomentosa (UT), Petiveria alliacea (PA) y Phyllantus niruri (PN), sobre el crecimiento y la metástasis de células de melanoma B16/BL6 en ratones C57BL/6, y sobre los niveles séricos del factor de necrosis tumoral alfa (TNF-iÀ), la interleucina-6 (IL-6) y el componente amiloide P (SAP). Ninguno de los tres extractos inhibió el crecimiento de las células B16/BL6, ni de las líneas celulares tumorales humanas, HT 29 y Caco-2, in vitro. Los niveles séricos de IL-6 y SAP, y la respuesta de TNF-¡À sérico a LPS resultaron elevados en los animales inoculados i.v. con las células de melanoma. Sólo el UT, inyectado i.p., redujo significativamente la metástasis a pulmón de un inóculo i.v. de células B16/BL6, y produjo un retardo en el crecimiento de un tumor primario. Además, sólo el extracto de UT produjo disminución de la respuesta sérica de las dos citocinas proinflamatorias, TNF-¡À e IL-6. UT amerita futuros estudios como potencial agente anticáncer.

Several useful anticancer drugs have been derived from plants. The growth and metastasis of tumours in the mousemay be used to study plant extracts with therapeutic potential. In the present study we investigated the possible effects of aqueous extracts of three Amazonian plants (Uncaria tomentosa [UT], Petiveria alliacea [PA] and Phyllantus niruri [PN]) on the growth and metastasis of B16/BL6 melanoma cells in the C57BL/6 mouse, and on the serum levels of tumor necrosis factor alpha (TNF-a), interleukin-6 (IL-6) and serum amyloid P component (SAP). None of the three extracts inhibited the growth of the B16/BL6 cells in vitro, or of two other tumor cell lines, HT-29 and Caco-2. Serum levels of IL-6 and SAP, and the TNF-¦Á serum response to LPS rose in the animals inoculated i.v. with melanoma cells. Only UT, when injected i.p. every other day for 14 days, was able to significantly reduce lung metastases after an i.v. inoculation of melanoma cells or to delay the appearance and growth of solid tumors after s.c. inoculation. In addition, only UT was able to reduce the serum response of the two proinflammatory cytokines, TNF-a and IL-6. No effect on SAP levels was observed. UT merits further study as an anticancer agent.

[Molecular and serological diagnosis of a dengue outbreak in Coro, Falcón state, Venezuela]. / Diagnóstico molecular y serológico de un brote de dengue en Coro, estado Falcón, Venezuela.

Dengue virus (DV) is responsible for a spectrum of diseases, from a self-limited fever disease (DF, dengue fever) to the more severe forms of hemorrhagic fever/dengue shock syndrome (DHF/DSS). The aim of this study was the serological and molecular confirmation of an outbreak of dengue in Falcon state, Venezuela. A total of 54 sera from patients with clinical diagnosis of DV infection were analyzed by an enzyme immunoassays developed in Venezuela (ELISA -IgM e -IgG) and by PCR. From them, 78% exhibited DV infection (PCR+ y/o IgM+), 48% exhibited viremia by PCR and 57% were positive to IgM. An interesting observation was the high percent (76%) of patients with past or secondary infection (IgG positive), which included all the patients exhibiting clinical symptoms of DHF (n = 8). From the PCR positive sera, serotype 1 was found in 27%, serotype 2 in 54% and serotype 4 in 19%. No serotype 3 was found circulating in this population, although this serotype was already circulating in the nearby island of Aruba. The combination of serological and molecular methods allow us to obtain a fairly precise information of this outbreak.

Diagnóstico molecular y serológico de un brote de dengue en Coro, estado Falcón-Venezuela / Molecular and serological diagnóstic of an outbreak of dengue in Coro, state Falcon-Venezuela

El virus dengue (VD) es responsable de un espectro de enfermedades que van desde una enfermedad febril autolimitante (FD, fiebre por dengue) hasta las formas severas, fiebre hemorrágica/síndrome de shock por dengue (FHD/SSD). El propósito del presente estudio fue la confirmación serológica y molecular de un brote de dengue en el Estado Falcón, Venezuela con la finalidad de confirmar la etiolgía de la enfermedad, determinar los serotipos infectantes y la relación del diagnóstico clínico con las respuestas inmunitarias en los pacientes con infección activa. Se analizaron 54 sueros de pacientes con diagnóstico clínico de infección por VD, mediante inmunoensayos enzimaticos desarrollados en Venezuela (ELISA, IgM e IgG) y por PCR. El 78 por ciento de los pacientes mostró evidencia de infección por VD (PCR+ y/o IgM), 48 por ciento presentaron viremia demostrada por PCR+ y 57 resultaron positivos a IgM, lo que sugiere que un alto número de los casos reportados como dengue en el país se deben efectivamente a la infección por VD. Un hecho resaltante fue el alto porcentaje (76 por ciento) de pacietnes con infección pasada o secundaria al VD (IgG positivos), dentro de los cuales se encontraban la totalidad de los pacientes diagnósticados clínicamente con FHD (n=8). De los pacientes PCR positivos, el 27 por ciento correspondió al serotipo 1,54 por ciento al serotipo 2 y 19 al serotipo 4. Para el momento de esta evaluación no se determinó la presencia del serotipo 3, aunque se conocía de su presencia en la cercana Isla de Aruba, la combinación de métodos serológicos y moleculares nos permitió obtener una información bastante precisa de este brote.

Distribucion anatomica de la grasa corporal subcutanea en una muestra escolar de Los Teques: patrones absolutos y agregación familiar / Anatomical distribution of subcutaneous body fat in a sample of school children of Los Tegues: absolute patterns and familial aggregation

En una muestra de la población escolar de la ciudad de Los Teques, Estado Miranda, Formada por 222 hembras y 216 varones, cuyas edades oscilaron entre los 6 y 17 años, ambas inclusive, se estudiaron los patrones absolutos de la distribución anatómica de la grasa corporal subcutánea. Estos fueron obtenidos en base a los promedios por grupo etario y sexo, de las medidas netas, expresadas en milímetros, de cinco pliegues de piel: tríceps, subescapular, suprailíaco, muslo y media pantorrilla. Igualmente, la semejanza entre hermanos para las variables talla, peso y los cinco pliegues antes mencionados, fue evaluada a través del coeficiente de correlación intraclase; para ello se identificaron un total de 100 pares de hermanos, y para hacer las comparaciones respectivas, un total de 100 pares de individuos no emparentados biológicamente. Los patrones absolutos muestran una mayor adiposidad total en el sexo femenino, y un desplazamiento de la grasa corporal, desde las extremidades hacia el tronco, lo cual ocurre desde las etapas cercanas a la dolescencia hasta las etapas cercanas a la adultez, siendo más evidente y precoz en el sexo masculino que en el femenino. Los coeficientes de correlación entre hermanos para todas las variables, son mayores que los correspondientes entre individuos no emparentados, a excepción del pliegue de la pantorilla. Las tendencias observadas señalan la influencia de factores genéticos y/o de ambiente común en la determinación de las mismas. La mayor correlación entre hermanos se observa para la talla y con respecto a los pliegues de piel, el muslo muestra la mayor correlación, mientras que la pantorrilla muestra la menor, sugiriendo que es la que tiene un menor componente genético en su determinación