930 resultados para Growth media for callus formation


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The study of quantum degenerate gases has many applications in topics such as condensed matter dynamics, precision measurements and quantum phase transitions. We built an apparatus to create 87Rb Bose-Einstein condensates (BECs) and generated, via optical and magnetic interactions, novel quantum systems in which we studied the contained phase transitions. For our first experiment we quenched multi-spin component BECs from a miscible to dynamically unstable immiscible state. The transition rapidly drives any spin fluctuations with a coherent growth process driving the formation of numerous spin polarized domains. At much longer times these domains coarsen as the system approaches equilibrium. For our second experiment we explored the magnetic phases present in a spin-1 spin-orbit coupled BEC and the contained quantum phase transitions. We observed ferromagnetic and unpolarized phases which are stabilized by the spin-orbit coupling’s explicit locking between spin and motion. These two phases are separated by a critical curve containing both first-order and second-order transitions joined at a critical point. The narrow first-order transition gives rise to long-lived metastable states. For our third experiment we prepared independent BECs in a double-well potential, with an artificial magnetic field between the BECs. We transitioned to a single BEC by lowering the barrier while expanding the region of artificial field to cover the resulting single BEC. We compared the vortex distribution nucleated via conventional dynamics to those produced by our procedure, showing our dynamical process populates vortices much more rapidly and in larger number than conventional nucleation.

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O crescimento celular da microalga Haematococcus pluvialis e a bioprodução de carotenoides são influenciados pelas diferentes condições de cultivo. Dentre os corantes naturais, a astaxantina tem importante aplicação farmacêutica, cosmética e na indústria de alimentos. Este pigmento além de colorir, possui propriedades biológicas, dentre elas a atividade antioxidante. A produção de astaxantina através do cultivo de H. pluvialis pode alcançar até 4% do peso seco da microalga. O objetivo desse trabalho foi avaliar o crescimento celular, bem como a produção de carotenoides pela microalga Haematococcus pluvialis em diferentes condições de cultivo e a atividade antioxidante dos extratos carotenogênicos. Foram utilizados os meios autotróficos Blue Green-11 (BG-11), BAR (Barbera Medium) e BBM (Bold Basal Medium) e os meios mixotróficos BBM e glicose e BBM e acetato de sódio, empregando 10 ou 20% de inóculo em pHs iniciais de 6, 7 ou 8, aeração de 0,30 L.min-1 , sob iluminância de 6 Klx, 24±1ºC durante 15 dias em fotobiorreatores de 1 L. A concentração celular foi avaliada diariamente através de leitura de absorvância a 560 nm. A ruptura celular foi realizada através de 0,05 g de células secas com 2 mL de dimetilsulfóxido e a concentração de carotenoides totais determinada a partir de leitura espectrofotométrica a 474 nm. Os meios de cultivo BG-11, BBM e glicose e BBM e acetato de sódio apresentaram, respectivamente, o maior crescimento celular e produção de carotenoides totais de 0,64, 1,18 e 0,68 g.L-1 , e 3026,66, 2623,12 e 2635,38 µg.g-1 , empregando 10% de inóculo em pH inicial de 7. Com base nesses resultados, foram selecionados esses três meios para dar continuidade ao trabalho. O meio de cultivo BBM e acetato de sódio obteve o melhor valor de concentração celular máxima, com 1,29±0,07 g.L-1 e de carotenoides totais 5653,56 µg.g-1 empregando pH inicial de 7 e concentração de inóculo de 20%. Este meio foi selecionado para a realização dos cultivos com injeção de 30 % de CO2 uma vez ao dia durante 1 hora, realizados durante 22 dias, em pH inicial de 7 e 20% de inóculo, com 30% de injeção de CO2 uma vez ao dia durante 1 hora. Nestas condições o crescimento celular alcançou o máximo de 1,13 g.L-1 (10 dias), carotenoides totais específicos de 2949,91 µg.g-1 e volumétricos de 764,79 µg.g-1 .L-1 (22 dias). A capacidade antioxidante dos extratos carotenogênicos também foi avaliada pelos métodos DPPH, FRAP e ABTS, não sendo possível quantificá-la através do DPPH e FRAP. Por outro lado, utilizando o método ABTS, em 90 minutos de reação, o poder de inibição encontrado foi de 35,70 % μg-1 . Assim, a condição que mais se destaca é a utilização do meio de cultivo BBM e acetato de sódio, com pH inicial 7, com 20% de inóculo, 0,30 L.min-1 de aeração, 6 Klx e 24±1ºC, uma vez que o crescimento celular e a bioprodução de carotenoides foi significativamente superior quando comparada às demais condições estudadas. Além disso, os carotenoides produzidos pela H. pluvialis, nesta condição, apresentaram capacidade antioxidativa.

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The most native fruit trees are belonging to Myrtaceae family, which it have as main marketing potential their fruit. Despite the wide acceptance of the fruits of these native fruit cultura, the establishment of commercial orchards is still necessary, because if it prevails extraction in the forest. To start the cultivo in the orchard, the first point is on the mother plant choice, which should provide superior characteristics when compared to other genotypes. Then, it is necessary to choose the method to can produce satisfactory amount of seedlings and preferably without it to lose the mother plant characteristics. For this, it adopts the asexual thechniques, with option for grafting, cuttings and air layering. These techniques when tested with native fruits tree, it proved limiting in theses results, with this, it should to test other it to recommend its use, especially, those fruit native of higher potential as jabuticaba tree, pitanga tree, sete capote tree and araça amarelo tree. The aim of this study was to test the use of asexual propagation through mini-cuttings in these native fruit trees, according to the time of collection, the mini-cutting length and concentration of IBA, as well as, it to relate the results of rooting with tryptophan extracted at certain times. The work was carried out at Universidade Tecnológica Federal do Paraná – Câmpus Dois Vizinhos, Brazil. The samples were collected each two months. The mini-cutting were prepared with 6 or 8 cm, with a pair of leaves reduced to 25% of the original size. The mini-cuttings had their base immersed in liquid solution of indole-butyric acid (IBA) in the concentrations of 0, 3000 and 6000 mg L-1 and then were placed in tubes containing commercial substrate. The experimental design was completely randomized with factorial 2 x 3 x 6 (mini-cutting length x IBA concentration x time of collection), with four replications, it being each plot varied according to the amount of shoots obtained by period time. After 120 days, the rooting and callus formation (%), average number of roots per mini-cutting and the average length of the roots were evaluated. After 60 days of these evaluations, the survival of mini-cuttings rooted after transplant was evaluated. It was evaluated also the production of mini-cuttings of each size in each period time. At the end of the experiment it was evaluated the percentage of survival of mother plantlets. For analysis of tryptophan was used materials branches, leaves and twigs with leaves, taken from the materials used for the production of mini-cutting. It was recommended for hybrid jabuticaba tree the use mini-cutting with eight cm, treated with 6000 mg L-1 of IBA and collected in June. For jabuticaba tree of cabinho and araça amarelo tree the period for propagation by mini-cuttings should be in August, regardless of IBA concentration and length of the mini-cutting. In the jabuticaba tree sabara and sete capote tree is important to obtain more satisfactory results realized the collect in October or December, with the same independence of other levels tested in other factors. However, for sete capote tree should test other techniques to increase the efficiency of propagation. And with pitanga tree recommended to the collection in June, but with 6cm the application of 3000 mg L-1 of IBA and 8 cm with 6000 mg L-1 of IBA.

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We introduce an innovative approach to the simultaneous control of growth mode and magnetotransport properties of manganite thin films, based on an easy-to-implement film/substrate interface engineering. The deposition of a manganite seed layer and the optimization of the substrate temperature allows a persistent bi-dimensional epitaxy and robust ferromagnetic properties at the same time. Structural measurements confirm that in such interface-engineered films, the optimal properties are related to improved epitaxy. A new growth scenario is envisaged, compatible with a shift from heteroepitaxy towards pseudo-homoepitaxy. Relevant growth parameters such as formation energy, roughening temperature, strain profile and chemical states are derived.

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O Programa de Aceleração do Crescimento (PAC) abrange uma série de medidas de caráter econômico, e teve como finalidade promover a retomada dos investimentos em setores estratégicos da economia. Este trabalho teve como objetivo principal comparar um período sem o programa (1994-2006) com o período em que foi implantado o PAC (2007-2013). Assim, pode-se verificar que o crescimento da economia brasileira, medido através do Produto Interno Bruto (PIB) a preços correntes, foi influenciado pelos investimentos ocorridos através do programa, mesmo que esse intervalo seja marcado por uma grande crise, em 2008. Pois, como apontam os resultados obtidos desse estudo, no intervalo 2007-2013 as taxas médias de crescimento do PIB, Formação Bruta de Capital Fixo (FBCF) e Utilização da Capacidade Instalada (UCI) são maiores do que as do intervalo 1994- 2006.

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Introducción El motivo principal por el que acuden los pacientes a las consultas de podología es el dolor producido por los callos y callosidades plantares. El dolor producido por las callosidades y callos plantares provocan en el paciente cambios de presiones y alteraciones en el apoyo, dificultando la deambulación correcta. Existen numerosos estudios sobre la eliminación de callosidades en pacientes diabéticos, con AR, pero pocos en personas sanas. La eliminación de estos callos y callosidades se puede realizar mediante deslaminación mecánica con bisturí o mediante queratolíticos. Objetivos Objetivo principal Analizar el efecto de la deslaminación mecánica con bisturí de las callosidades y callos plantares sobre el dolor y la calidad de vida en sujetos sanos Objetivos secundarios Determinar la existencia de modificaciones en los parámetros de la marcha con la eliminación de callosidades y callos plantares Observar las diferencias y efectividad de tratamientos de la eliminación de callosidades plantares mediante la técnica de deslaminación mecánica con bisturí versus parches de ácido salicílico Comprobar los cambios producidos en los parámetros psíquicos y fiscos del paciente antes y después de las diferentes técnicas de eliminación de las callosidades empleadas Método Se realizan dos estudios: un estudio cuasi experimental aleatorizado no controlado, en el que a un grupo de 34 pacientes con callosidades plantares dolorosas se les mide el dolor con una escala visual analógica y para analizar los parámetros de la marcha, la paltaformaWin-Track, antes del tratamiento de deslaminación mecánica con bisturí y a las 24 horas. El segundo estudio es un ensayo clínico aleatorizado inscrito en Australian New ZelandClinicalstrials y aprobado por el Comité ético de la Universidad de Málaga, en el que 62 participantes con callosidades plantares dolorosas se dividieron en dos grupos de tratamiento. El grupo A recibió tratamiento con parche de ácido salicílico y el grupo B recibió tratmiento de deslaminación con bisturí. Se utilizó la escala visual analógica para la medida de dolor antes, inmediatamente después de la intervención, a las 2 semanas y a las 6 semanas. Para el dolor y la discapacidad funcional del pie se utilizó el cuestionario Manchester FootPain and Disability antes del tratamiento, a las 2 semanas y a las 6 semanas. Para medir la calidad de vida general se utilizó el cuestionario SF-12 Conclusiones La deslaminación mecánica con bisturí de los callos y callosidades plantares es efectiva para su eliminación a nivel de la sensación de dolor, aunque no tanto en lo que se refiere a la mejora de calidad de vida. No hay resultados significativos de que la eliminación mecánica con bisturí de callos y callosidades plantares modifican los parámetros de la marcha medido con la plataforma Win-track. Se observa como la deslaminación mecánica con bisturí para la eliminación de callos y callosidades plantares pueden ser más efectiva a corto plazo que la eliminación mediante parche con ácido salicílico. Se observa cómo se modifica los paramentos psíquicos en el grupo de tratamiento con parche con ácido salicílico, aunque con una significación baja. Bibliografía Balanowski, K. R., & Flynn, L. M. (2005). Effect of painful keratoses debridement on foot pain, balance and function in older adults. Gait & Posture, 22(4), 302-307. http://doi.org/10.1016/j.gaitpost.2004.10.006 Collins, S. L., Moore, R. A., &McQuay, H. J. (1997). The visual analogue pain intensity scale: what is moderate pain in millimetres? Pain, 72(1-2), 95-97. Coughlin, M. J. (2000).Common Causes of Pain in the Forefoot in Adults. Journal of Bone & Joint Surgery, British Volume, 82-B(6), 781-790. Farndon, L. J., Vernon, W., Walters, S. J., Dixon, S., Bradburn, M., Concannon, M., & Potter, J. (2013). The effectiveness of salicylic acid plasters compared with «usual» scalpel debridement of corns: a randomised controlled trial. Journal of Foot and Ankle Research, 6(1), 40. http://doi.org/10.1186/1757-1146-6-40 Freeman, D. B. (2002). Corns and calluses resulting from mechanical hyperkeratosis. American FamilyPhysician, 65(11), 2277-2280. Gijon-Nogueron, G., Ndosi, M., Luque-Suarez, A., Alcacer-Pitarch, B., Munuera, P. V., Garrow, A., & Redmond, A. C. (2014). Cross-cultural adaptation and validation of the Manchester Foot Pain and Disability Index into Spanish. Quality of Life Research: An International Journal of Quality of Life Aspects of Treatment, Care and Rehabilitation, 23(2), 571-579. http://doi.org/10.1007/s11136-013-0507-5 Grouios, G. (2005). Footedness as a potential factor that contributes to the causation of corn and callus formation in lower extremities of physically active individuals. The Foot, 15(3), 154-162. http://doi.org/10.1016/j.foot.2005.05.003 Landorf, K. B., Morrow, A., Spink, M. J., Nash, C. L., Novak, A., Potter, J., &Menz, H. B. (2013). Effectiveness of scalpel debridement for painful plantar calluses in older people: a randomized trial. Trials, 14, 243. http://doi.org/10.1186/1745-6215-14-243 Lang, L. M. G., Simmonite, N., West, S. G., & Day, S. (1994). Salicylic acid in the treatment of corns. The Foot, 4(3), 145-150. http://doi.org/10.1016/0958-2592(94)90019-1 Luo, X., Lynn George, M., Kakouras, I., Edwards, C. L., Pietrobon, R., Richardson, W., & Hey, L. (2003). Reliability, validity, and responsiveness of the short form 12-item survey (SF-12) in patients with back pain. Spine, 28(15), 1739-1745. http://doi.org/10.1097/01.BRS.0000083169.58671.96 Ramachandra, P., Maiya, A. G., & Kumar, P. (2012). Test-retest reliability of the Win-Track platform in analyzing the gait parameters and plantar pressures during barefoot walking in healthy adults. Foot & Ankle Specialist, 5(5), 306-312. http://doi.org/10.1177/1938640012457680 Siddle, H. J., Redmond, A. C., Waxman, R., Dagg, A. R., Alcacer-Pitarch, B., Wilkins, R. A., &Helliwell, P. S. (2013). Debridement of painful forefoot plantar callosities in rheumatoid arthritis: the CARROT randomised controlled trial. Clinical Rheumatology, 32(5), 567-574. http://doi.org/10.1007/s10067-012-2134-x

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Due to the accelerating processes of soil salinization and shortage of fresh water, the practice of saline agriculture is gaining momentum in many areas of the world. However, there are some concerns that using saline water for irrigation may be non-environmentally sustainable, with potential to cause irreversible soil degradation. In addition, there is a lack of information on the morphological, physiological, and biochemical changes that can occur in plants when irrigated with saline water. In light of the above, the major aim of this work was to investigate the effects of a range of water salinity levels and irrigation regimes on the performances of salt tolerant species promising as future crop plants for saline agriculture. The following objectives were addressed: To determine the effects of different water regimes (leaching irrigation vs. no leaching irrigation) with water at increasing salinity concentrations on the growth, ion accumulation and water relations of Sorghum bicolor plants grown under saline soil conditions. To describe the germination response of Salicornia europaea seeds across a wide range of water salinity levels through six reliable indices for screening salinity tolerance at the seed germination stage. To explore the different physiological responses of six wild halophytes commonly found in the Mediterranean area (Artemisia absinthium, Artemisia vulgaris, Atriplex halimus, Chenopodium album, Salsola komarovii, and Sanguisorba minor), and rank their tolerance after exposure to growing levels of water salinity. To identify the main adaptation mechanisms that distinguish C3 from C4 halophytes when exposed to increasing salinity in the growth media, through a comparative study between the C3 species Atriplex hortensis and the C4 species Atriplex halimus. To identify the main adaptation mechanisms that distinguish annual from perennial halophytes when exposed to severe conditions of salinity and drought, through a comparative analysis between two annual Salicornia spp. and the perennial Sarcocornia fruticosa.

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The aim of the present paper was to evaluate cyst formation and growth parameters of Borrelia garinii in a range of media differing in formulation and cost. A qualitative assessment of morphology and motility of B. garinii was conducted. All media were prepared aseptically and used in test tubes or Petri dishes. For each medium, the initial spirochete concentration was standardized to 10³ spirochets/mL. The following culture media were suitable to grow B. garinii: Barbour-Stoenner-Kelly, brain heart infusion and PMR. Growth was minimal at six weeks post-inoculation and maximum spirochete density was observed between 9-12 weeks. Often, the cultures developed cysts of different sizes, isolated or in groups, with a spiraled portion of variable sizes, mainly in unfavorable culture media. Brazilian Lyme disease-like illness, also known as Baggio-Yoshinari syndrome (BYS), is a new and interesting emerging tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, only during its cystic forms. It has been assumed that the peculiar clinical and laboratory features of BYS are consequential to the absence of a human sucker Ixodes ricinus complex tick at risk areas in Brazil, supporting the concept that the borrelia phenotypic expression pattern is modified as it is transmitted through the host.

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Tillandsia gardneri Lindi, is a herbaceous perennial with ornamental value. However, in Brazil there is no report about this species' cultivation on a commercial scale. The low multiplication rate of T. gardneri (in average one offshoot/plant/year) leads to illegal over-collection in the wild to meet commercial demands. The development of protocols for in vitro propagation of T. gardneri may be useful for increasing multiplication rate, producing enough plants to supply the ornamental market and also to reduce the pressure over plants collection in the wild. The present study evaluated the effect of growth regulators (6-benzylaminopurine-BA alone or in combinations with naphthaleneacetic acid-NAA) on shoots development from seedlings pre-established in vitro, from seed germination on 1/4 MS medium without growth regulators. Seedlings (with about 1.0 cm long) were re-cultured to solid 1/2 MS media supplemented with growth regulators. After 30 days on the induction medium seedlings were re-cultured to MS basal medium. The experiment was conducted in a complete randomized design with four replications and ten treatments: control (free of growth regulators), BA (0.5, 1.0 and 2.0 mg/L), BA (0.5 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.5 mg/L), BA (2.0 mg/L) + ANA (0.1 mg/L), BA (2.0 mg/L) + ANA (0.5 mg/L), and BA (2.0 mg/L) + ANA (1.0 mg/L). The outgrowth of shoots did not occur on medium devoid of growth regulators (control). Regression analysis for some evaluated parameters, such as percentage of seedlings responsive to shoot formation and number of shoots/seedling, and regulators concentrations (BA or ANA) were significant, allowing the establishment of the growth regulators concentration for obtaining the best multiplication rate. Some seedlings maintained in media with ANA (0.5 or 1.0 mg/L) were completely converted into callus masses that turned dark brown leading to seedlings death.

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Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.

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In vitro culture of the mutualistic fungus of leaf-cutting ants is troublesome due to its low growth rate, which leads to storage problems and contaminants accumulation. This paper aims at comparing the radial growth rate of the mutualistic fungus of Atta sexdens rubropilosa Forel in two different culture media (Pagnocca B and MEA LP). Although total MEA LP radial growth was greater all along the bioassay, no significant difference was detected between growth efficiencies of the two media. Previous evidences of low growth rate for this fungus were confirmed. Since these data cannot point greater efficiency of one culture medium over the other, MEA LP medium is indicated for in vitro studies with this mutualistic fungus due its simpler composition and translucent color, making the analysis easier.

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Crude extracts of a callus culture (two culture media) and adult plants (two collections) from Alternanthera tenella Colla (Amaranthaceae) were evaluated for their antibacterial and antifungal activity, in order to investigate the maintenance of antimicrobial activity of the extracts obtained from plants in vivo and in vitro. The antibacterial and antifungal activity was determined against thirty strains of microorganisms including Gram-positive and Gram-negative bacteria, yeasts and dermatophytes. Ethanolic and hexanic extracts of adult plants collected during the same period of the years 1997 and 2002 [Ribeirao Preto (SP), collections 1 and 2] and obtained from plant cell callus culture in two different hormonal media (AtT43 and AtT11) inhibited the growth of bacteria, yeasts and dermatophytes with inhibition halos between 6 and 20 mm. For the crude extracts of adult plants bioassay-guided fractionation, purification, and isolation were performed by chromatographic methods, and the structures of the isolated compounds were established by analysis of chemical and spectral evidences (UV, IR, NMR and ES-MS). Steroids, saponins and flavonoids (aglycones and C-glycosides) were isolated. The minimum inhibitory concentration (MIC) of the isolated compounds varied from 50 to 500 mu g/mL.

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Resveratrol is a stilbene compound found in grapes and other sources. In this study we examined the effects of trans-resveratrol (4.38-438 mu M/implant) in the vasculogenesis of yolk-sac membranes and its capacity to improve chick embryo growth. High concentrations of the stilbene (43.8-438 mu M) significantly inhibited early vessel formation, decreasing the percentage vitelline vessels of 3.5-day embryos by 50% compared to the control. In addition, basic fibroblast growth factor-stimulated vasculogenesis (140% of vessels as compared to control) was partially reversed by t-resveratrol (35% of inhibition) and treatments with cyclooxygenase inhibitors (acetylsalicylic acid and indomethacin) as well a protein-kinase C (PKC) activator (phorbol-12,13-dibutyrate) decreased the vessel number to 60%, 50%, and 44%, respectively. Treatments with t-resveratrol (4.38-43.8 mu M/implant) significantly increased the body length of embryos incubated in vitro uncoupled from any impairment in the body shape or detectable embryotoxic effect. We suggest that the antivasculogenic activity and the enhancement in embryonic growth promoted by non acute treatments with t-resveratrol were, at least in part, due to PKC inhibition. We suggest that t-resveratrol can be usable not only as a reliable functional nutriment, but also is useful for the development of prophylactic and/or therapeutic agents for treatment of angiogenic-degenerative diseases.

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Despite wide clinical application, the efficacy of platelet-rich plasma (PRP) for repairing bone defects and enhancing osseointegration of metal implants is still subject of debate. This study aimed to evaluate the effects of a well-defined PRP-like mixture containing platelet-derived growth factor-BB, transforming growth factor (TGF)-beta 1, TGF-beta 2, albumin, fibronectin, and thrombospondin [growth factors (GFs) + proteins] on the development of the osteogenic phenotype on titanium (Ti) in vitro. Human alveolar bone-derived osteoblastic cells were subcultured on Ti discs and exposed during the first 7 days to osteogenic medium supplemented with GFs + proteins and to osteogenic medium alone thereafter up to 14 days. Control cultures were exposed to only osteogenic medium. Dose-response experiments were carried out using rat primary calvarial cells exposed to GFs + proteins and 1:10 or 1:100 dilutions of the mixture. Treated human-derived cell cultures exhibited a significantly higher number of cycling cells at days 1 and 4 and of total cells at days 4 and 7, significantly reduced alkaline phosphatase (ALP) activity at days 4, 7, and 10, and no Alizarin red-stained areas (calcium deposits) at day 14, indicating an impairment in osteoblast differentiation. Although the 1:10 and 1:100 dilutions of the mixture restored the proliferative activity of rat-derived osteogenic cells to control levels and promoted a significant increase in ALP activity at day 10 compared with GFs + proteins, mineralized nodule formation was only observed with the 1:100 dilution (similar to 50% of the control). These results showed that a PRP-like protein mixture inhibits development of the osteogenic phenotype in both human and rat osteoblastic cell cultures grown on Ti. (J Histochem Cytochem 57:265-276, 2009)

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The role of nitrate, ammonium, and culture medium pH on shoot organogenesis in Nicotiana tabacum zz100 leaf discs was examined. The nitrogen composition of a basal liquid shoot induction medium (SIM) containing 39.4 mM NO3- and 20.6 mM NH4+ was altered whilst maintaining the overall ionic balance with Na+ and Cl- ions. Omission of total nitrogen and nitrate, but not ammonium, from SIM prevented the initiation and formation of shoots. When nitrate was used as the sole source of nitrogen, a high frequency of explants initiated and produced leafy shoots. However, the numbers of shoots produced were significantly fewer than the control SIM. Buffering nitrate-only media with the organic acid 2[N-morpholinol]thanesulphonic acid (MES) could not compensate for the omission of ammonium. Ammonium used as the sole source of nitrogen appeared to have a negative effect on explant growth and morphogenesis, with a significant lowering of media pH. Buffering ammonium-only media with MES stabilized pH and allowed a low frequency of explants to initiate shoot meristems. However, no further differentiation into leafy shoots was observed. The amount of available nitrogen appears to be less important than the ratio between nitrate and ammonium. Shoot formation was achieved with a wide range of ratios, but media containing 40 mM nitrate and 20 mM ammonium (70:30) produced the greatest number of shoots per explant. Results from this study indicate a synergistic effect between ammonium and nitrate on shoot organogenesis independent of culture medium pH.