In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l(-1) IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time-and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

Dispersible crystalline nanobundles of YPO4 and Ln (Eu, Tb)-doped YPO4: rapid synthesis, optical properties and bio-probe applications

Undoped and Ln(3+) (Eu and Tb)-doped crystalline nanobundles of YPO4 were prepared by a facile microwave-assisted route with water as a solvent and without using any surfactant. TEM investigations reveal that the as-prepared powder consists of lenticular-shaped nanobundles (similar to 100 nm in diameter) made of very small nanorods with diameter less than 10 nm and length varying from 20 to 50 nm. Each nanorod in turn is single crystalline, as revealed by HRTEM imaging. The as-prepared nanobundles are easily dispersible in various solvents, especially water, without any surface functionalization, which is critical for various bio-probe applications like cell and tissue imaging. The Eu- and Tb-doped YPO4 nanobundles show good photoluminescence properties and were further evaluated for their use as fluorescent biolabels. Our results show that HeLa cells labelled with Eu- and Tb-doped YPO4 nanobundles show bright red (Eu) and green (Tb) intracellular luminescence under a confocal microscope. Concentration-and time-dependent MTT cell viability assays show that the nanobundles show low toxicity towards cells which makes them promising in bioimaging field.

An Endophytic Fungus, Talaromyces radicus, Isolated from Catharanthus roseus, Produces Vincristine and Vinblastine, Which Induce Apoptotic Cell Death

Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS) region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp-CrP14, obtained from stem tissues, and Talaromyces radicus-CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 mu g/ml and 20 mu g/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC), the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus-CrP20 (NCBI GenBank accession number KC920846). The production of vincristine and vinblastine by T. radicus-CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 mu g/l) in modified M2 medium and of vinblastine (70 mu g/l) in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns.

Combined Effect of Cryogel Matrix and Temperature-Reversible Soluble-Insoluble Polymer for the Development of in Vitro Human Liver Tissue

Hepatic cell culture on a three-dimensional (3D) matrix or as a hepatosphere appears to be a promising in vitro biomimetic system for liver tissue engineering applications. In this study, we have combined the concept of a 3D scaffold and a spheroid culture to develop an in vitro model to engineer liver tissue for drug screening. We have evaluated the potential of poly(ethylene glycol)-alginate-gelatin (PAG) cryogel matrix for in vitro culture of human liver cell lines. The synthesized cryogel matrix has a flow rate of 7 mL/min and water uptake capacity of 94% that enables easy nutrient transportation in the in vitro cell culture. Youngs modulus of 2.4 kPa and viscoelastic property determine the soft and elastic nature of synthesized cryogel. Biocompatibility of PAG cryogel was evaluated through MTT assay of HepG2 and Huh-7 cells on matrices. The proliferation and functionality of the liver cells were enhanced by culturing hepatic cells as spheroids (hepatospheres) on the PAG cryogel using temperature-reversible soluble-insoluble polymer, poly(N-isopropylacrylamide) (PNIPAAm). Pore size of the cryogel above 100 mu m modulated spheroid size that can prevent hypoxia condition within the spheroid culture. Both the hepatic cells have shown a significant difference (P < 0.05) in terms of cell number and functionality when cultured with PNIPAAm. After 10 days of culture using 0.05% PNIPAAm, the cell number increased by 11- and 7-fold in case of HepG2 and Huh-7 cells, respectively. Similarly, after 10 days of hepatic spheroids culture on PAG cryogel, the albumin production, urea secretion, and CYP450 activity were significantly higher in case of culture with PNIPAAm. The developed tissue mass on the PAG cryogel in the presence of PNIPAAm possess polarity, which was confirmed using F-actin staining and by presence of intercellular bile canalicular lumen. The developed cryogel matrix supports liver cells proliferation and functionality and therefore can be used for in vitro and in vivo drug testing.

Stimuli-responsive protamine-based biodegradable nanocapsules for enhanced bioavailability and intracellular delivery of anticancer agents

Enzyme-and pH-responsive polyelectrolyte nanocapsules having diameters in the range of 200 +/- 20 nm were fabricated by means of Layer-by-Layer assembly of biopolymers, protamine, and heparin, and then loaded with anticancer drug doxorubicin. The incorporation of the FDA-approved peptide drug protamine as a wall component rendered the capsules responsive to enzyme stimuli. The stimuli-responsive drug release from these nanocapsules was evaluated, and further modulation of capsule permeability to avoid premature release was demonstrated by crosslinking the wall components. The interaction of the nanocapsules with cancer cells was studied using MCF-7 breast cancer cells. These capsules were readily internalized and disintegrated inside the cells, culminating in the release of the loaded doxorubicin and subsequent cell death as observed by confocal microscopy and MTT Assay. The bioavailability studies performed using BALB/c mice revealed that the encapsulated doxorubicin exhibited enhanced bioavailability compared to free doxorubicin. Our results indicate that this stimuli-responsive system fabricated from clinically used FDA-approved molecules and exhibiting minimal premature release has great potential for drug-delivery applications.

Stimuli-responsive protamine-based biodegradable nanocapsules for enhanced bioavailability and intracellular delivery of anticancer agents

Enzyme-and pH-responsive polyelectrolyte nanocapsules having diameters in the range of 200 +/- 20 nm were fabricated by means of Layer-by-Layer assembly of biopolymers, protamine, and heparin, and then loaded with anticancer drug doxorubicin. The incorporation of the FDA-approved peptide drug protamine as a wall component rendered the capsules responsive to enzyme stimuli. The stimuli-responsive drug release from these nanocapsules was evaluated, and further modulation of capsule permeability to avoid premature release was demonstrated by crosslinking the wall components. The interaction of the nanocapsules with cancer cells was studied using MCF-7 breast cancer cells. These capsules were readily internalized and disintegrated inside the cells, culminating in the release of the loaded doxorubicin and subsequent cell death as observed by confocal microscopy and MTT Assay. The bioavailability studies performed using BALB/c mice revealed that the encapsulated doxorubicin exhibited enhanced bioavailability compared to free doxorubicin. Our results indicate that this stimuli-responsive system fabricated from clinically used FDA-approved molecules and exhibiting minimal premature release has great potential for drug-delivery applications.

Thermal co-reduction approach to vary size of silver nanoparticle: its microbial and cellular toxicology

In recent years, silver nanoparticles (AgNPs) have attracted considerable interest in the field of food, agriculture and pharmaceuticals mainly due to its antibacterial activity. AgNPs have also been reported to possess toxic behavior. The toxicological behavior of nanomaterials largely depends on its size and shape which ultimately depend on synthetic protocol. A systematic and detailed analysis for size variation of AgNP by thermal co-reduction approach and its efficacy toward microbial and cellular toxicological behavior is presented here. With the focus to explore the size-dependent toxicological variation, two different-sized NPs have been synthesized, i.e., 60 nm (Ag60) and 85 nm (Ag85). A detailed microbial toxicological evaluation has been performed by analyzing minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), diameter of inhibition zone (DIZ), growth kinetics (GrK), and death kinetics (DeK). Comparative cytotoxicological behavior was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. It has been concluded by this study that the size of AgNPs can be varied, by varying the concentration of reactants and temperature called as ``thermal co-reduction'' approach, which is one of the suitable approaches to meet the same. Also, the smaller AgNP has shown more microbial and cellular toxicity.

Evaluación de nuevas variedades de tomate de consumo fresco (Lycopersicon esculentum Mill) en el Valle de Sébaco, Matagalpa

El trabajo se realizó en el periodo comprendido entre Julio y Octubre de 1995, en 1 a Estación Experimental "Raúl González" del Valle de Sebaco, Matagalpa. Se evaluaron nueve cultivares de tomate Lycopersicon esculentum MIJI), para consumo fresco, con el objetivo de comparar los resultados en cuanto a crecimiento, desarrollo y rendimiento. El ANDEVA realizado a los cultivares evaluados demostró que la mayoría de las variables medidas,diferenciaron a las distintas variedades estudiadas. En cuanto a floración y fructificación la mayor precocidad la presentaron los cultivares Topacio a los 19 y 25 días después del trasplante (ddt) y MTT-019 a los 20 y 27 ddt. Respecto a los rendimientos comerciales, el cultivar MTT-019 produjo los más altos valores con 9546 kg/ha, además mostro el mayor rendimiento total con 19764 kg/ha; las mayores pérdidas las presentó el cultivar Caribe con 72% respecto al rendimiento total. La matriz de correlación realizado, se obtuvo relación desde el punto de vista fenotípico entre los caracteres altura de planta con diámetro del tallo, número de racimos, número de Jóculos y grosor del mesocarpio; también se presentaron correlaciones positivas y altamente significativas entre el número de lóculos y los caracteres grosor del mesocarpio, diámetro polar y diámetro ecuatorial. La correlación entre los componentes del rendimiento mostró correlaciones positivas entre el rendimiento total y los caracteres número de plantas cosechadas, números de frutos sanos, numero de frutos desechados, rendimiento comercial, número total de frutos cosechados, pérdidas y número de racimos.

Evaluación del comportamiento de cinco materiales de tomate (Lycopersicum esculentum Mill) ante el ataque del complejo mosca blanca -geminivirus- en la región central del pais

El objetivo de este trabajo fue evaluar cinco materiales de tomate (Lycopersicum esculentum Milll) ante infestaciones natura les del complejo mosca blanca- geminivirus en la región Central del país en el período comprendido de Diciembre 2002 – Abril 2003. En este experimento se evaluaron las líneas TY-4, TY-12, TY-13 y las variedades MTT-013 y MTT-019, el diseño utilizado fue el de Bloques Completo al Azar (BCA). Las variables evaluadas fueron: población de moscas blancas/pta, incidencia de la enfermedad, severidad de la enfermedad, número de racimos florales, número de flores, número de frutos por planta y rendimientos (kg/ha). A los datos obtenidos se les aplicó un Análisis de Varianza y una separación de medias según Tukey. Los resultados indicaron que la línea TY-4 y la TY-13 fueron las más tolerantes al complejo mosca blanca – geminivirus, siendo la variedad MTT-013 la más susceptible. En cuanto a la severidad, la mejor línea es la TY-4 seguida de la TY-13 y la TY-12, siendo la más severamente afectada la variedad MTT-013. En el caso de la población de mosca blanca, la variedad MTT-013 fue en la que se encontró el mayor número de adultos y en la que se encontraron el menor número de adultos de mosca blanca fue la línea TY-13. Para las variables racimos florales y flores la variedad que obtuvo el mayor promedio fue la variedad MTT-013 y el menor la línea TY-4. En el caso de los frutos, el material con menor promedio fue la variedad MTT-013 y los mayores promedios los alcanzaron las líneas TY-12 y TY-13, respectivamente. El mayor rendimiento lo alcanzó la línea TY-13 con un rendimiento equivalente de 19,000 kg/ha y el menor la variedad MTT-013 con un rendimiento equivalente de 7,000 kg/ha. En la etapa de laboratorio se comprobó efectivamente la presencia de geminivirus en todos los materiales evaluados.

Rubens Paiva

Apresenta ensaio biográfico, discursos, cronologia, fotografias e documentos sobre a trajetória pessoal e política de Rubens Paiva.

Os Guayanazes : conto historico sobre a fundação de S. Paulo : recordação das ferias de 1858 a 1859

O autor esclarece, no prefácio da obra, ter sido ela redigida "aos trambolhos e às carreiras" e tratar-se de uma experiência num gênero novo - o ensaio. Mas J. Navarro de Andrade, na introdução de seu texto "Viagem ao Araguaia" diz o seguinte de José Vieira Couto de Magalhães : despretensioso, é, contudo, o conto aludido, um trabalho de merecimento, pena é que o autor não produzisse nesse sentido outros escritos, e seguisse outra vereda da vida.

Cartas do P. Antonio Vieira da companhia de Jesus

Coleção formada por três volumes, datados de 1735-1746, que contém, respectivamente, 141, 145 e 196 cartas escritas pelo Padre Vieira, que refletem a mais pura prosa da língua portuguesa. Segundo Francisco Freire de Carvalho, no seu Ensaio da história literária de Portugal, "estas cartas têm merecido ser emparelhadas em virtudes de estilo e na pureza de linguagem às de Cícero, ou pouco menos: e, como tais, elogiadas por todos quantos se prezam de bom gosto literário".

The Mitogenic and Anti-Apoptotic Activity of Tumor Conditioned Medium on Endothelium

This study was designed to observe the effect of tumor conditioned medium (TCM) on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to TCM from breast carcinoma cell line MDA-MB-231, then we measured their proliferation, apoptosis and cell cycle distribution by MTT and flow cytometery (FCM). Following the stimulation of TCM, HUVECs showed higher pro-mitogenic and anti-apoptotic ability than did the negative control group (ECGF-free medium with 20% FBS), but a similar ability to the positive control group (medium with ECGF and 20% FBS). From these results, we can conclude that breast carcinoma cell line MDA-MB-231 could secret soluble pro-angiogenic factors that induce HUVEC angiogenic switching, including cell cycle progression, proliferation and growth. The role and character of these factors remain to be further studied.

Marquês de Paranaguá

Ensaio biográfico de Chico Castro que resgata um dos políticos brasileiros mais importantes do II Reinado, mostrando que a pesquisa histórico-política ainda tem muito a contribuir para o entendimento da realidade atual de nossas instituições.