Immunoexpression of aromatase and estrogen receptors β in stem spermatogonia of bullfrogs indicates a role of estrogen in the seasonal spermatogonial mitotic activity

Bullfrog stem spermatogonia, also named primordial germ cells (PGCs), show strong testosterone immunolabeling in winter, but no or weak testosterone immunoexpression in summer. Thus, the role of testosterone in these cells needs to be clarified. In this study, we proposed to evaluate whether PGCs express aromatase and estrogen receptors, and verify a possible role of estrogen in PGCs seasonal proliferation. Testes of male adult bullfrogs, collected in winter (WG) and summer (SG), were fixed and embedded in historesin, for quantitative analysis, or paraffin for immunohistochemistry (IHC). The number of haematoxylin/eosin stained PGCs/lobular area was obtained. Proliferating cell nuclear antigen (PCNA), aromatase, estrogen receptor β (ERβ) and PCNA/ERβ double immunolabeling were detected by IHC. The number of PCNA-positive PGCs and the histological score (HSCORE) of aromatase and ERβ immunolabeled PGCs were obtained. Although the number of PGCs increased significantly in WG, a high number of PCNA-positive PGCs was observed in summer. Moreover, aromatase and ERβ HSCORE was higher in SG than WG. The results indicate that PGCs express a seasonal proliferative activity; the low mitotic activity in winter is related to the maximal limit of germ cells which can be supported in the large lobules. In SG, the increased ERβ and aromatase HSCORE suggests that testosterone is converted into estrogen from winter to summer. Moreover, the parallelism between the high PGCs mitotic activity and ERβ immunoexpression suggest a participation of estrogen in the control of the PGCs seasonal proliferative activity which guarantee the formation of new germ cysts from summer to next autumn. © 2012 Elsevier Inc.

Lipopolysaccharide-induced Stem Cell Factor Messenger RNA Expression and Production in Odontoblast-like Cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro study of CD133 human stem cells labeled with superparamagnetic iron oxide nanoparticles

Superparamagnetic iron oxide nanoparticles (SPIONs) are applied in stem cell labeling because of their high magnetic susceptibility as compared with ordinary paramagnetic species, their low toxicity, and their ease of magnetic manipulation. The present work is the study of CD133(+) stem cell labeling by SPIONs coupled to a specific antibody (AC133), resulting in the antigenic labeling of the CD133+ stem cell, and a method was developed for the quantification of the SPION content per cell, necessary for molecular imaging optimization. Flow cytometry analysis established the efficiency of the selection process and helped determine that the CD133 cells selected by chromatographic affinity express the transmembrane glycoprotein CD133. The presence of antibodies coupled to the SPION, expressed in the cell membrane, was observed by transmission electron microscopy. Quantification of the SPION concentration in the marked cells using the ferromagnetic resonance technique resulted in a value of 1.70 x 10 (13) mol iron (9.5 pg) or 7.0 x 10 (6) nanoparticles per cell ( the measurement was carried out in a volume of 2 mu L containing about 6.16 x 10 5 pg iron, equivalent to 4.5 x 10 (11) SPIONs). (c) 2008 Elsevier B.V. All rights reserved.

Low power laser radiation at 685 nm stimulates stem-cell proliferation rate in Dugesia tigrina during regeneration

Today's scientific interest in tissue engineering for organ transplantations and regeneration from stem cells, allied with recent observations on biostimulation of tissues and cells by laser radiation, stands as a strong motivation for the present work, in which we examine the effects of the low power laser radiation onto planarians under regenerative process. To investigate those effects, a number of 60 amputated worms were divided in three study groups: a control group and two other groups submitted to daily 1 and 3 min long laser treatment sections at similar to 910 W/m(2) power density. A 685 nm diode laser with 35 mW optical power was used. Samples were sent to histological analysis at the 4th, the 7th and the 15th (lays after amputation. A remarkable increase in stem cells counts for the fourth day of regeneration was observed when the regenerating worms was stimulated by the laser radiation. Our findings encourage further research works on the influence of optical radiation onto stem cells and tissue regeneration. (c) 2005 Elsevier B.V. All rights reserved.

Stem anatomy and development of successive cambia in Hebanthe eriantha (Poir.) Pedersen: A neotropical climbing species of the Amaranthaceae

Hebanthe eriantha (Poir.) Pedersen, a climbing species of the Amaranthaceae increases in stem thickness by forming successive cambia. The family is dominated by herbaceous species and is constantly under discussion due to its disputed nature of the meristem. In the young stem small alternate segments of vascular cambium cease to divide and new arc of cambium initiates outside to it. The newly formed arcs connect with pre-existing alternate segments of cambium to complete the ring. On the contrary, in thick stems, instead of small segments, complete ring of cambium is replaced by new one. These new alternate segments/cambia originate from the parenchyma cells located outside to the phloem produced by previous cambium. Cambium is storied and exclusively composed of fusiform initials while ray cells remain absent at least in the early part of the secondary growth. However, large heterocellular rays are observed in 15-mm diameter stems but their frequency is much lower. In some of the rays, ray cells become meristematic and differentiate into radially arranged xylem and phloem elements. In fully grown plants, stems are composed of several successive rings of secondary xylem alternating with secondary phloem. Secondary xylem is diffuse-porous and composed of vessels, fibres, axial parenchyma while exceptionally large rays are observed only in the outermost regions of thick stems. Vessel diameter increases progressively from the centre towards the periphery of stems. Although the origin of successive cambia and composition of secondary xylem of H. eriantha remains similar to other herbaceous members of Amaranthaceae, the occurrence of relatively wider and thick-walled vessels and large rays in fully grown plants is characteristic to climbing habit. © 2013 Springer-Verlag Wien.

Increased carbon dioxide concentration in the air reduces the severity of Ceratocystis wilt in Eucalyptus clonal plantlets

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Increased atmospheric carbon dioxide concentration: effects on eucalypt rust (Puccinia psidii), C:N ratio and essential oils in eucalypt clonal plantlets

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Measured and modeled interactive effects of potassium deficiency and water deficit on gross primary productivity and light-use efficiency in Eucalyptus grandis plantations

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos da tricostatina A sobre a acetilação de histonas, proliferação celular e diferenciação de células tronco embrionárias murinas

Background: Embryonic stem cells are cells derived from early-stage embryos that are characterized by pluripotency and self-renewal capacity. The in vitro cultured murine embryonic stem cells can indefinitely propagate in an undifferentiated state in the presence of leukemia inhibitory factor (LIF). However, when stimulated, these cells can differentiate into cell lines derived from all three embryonic germ layers. The trichostatin A (TSA) is an epigenetic modifier agent and several studies have used the TSA to stimulate cellular differentiation. However, most of these studies only assessed one TSA concentration. Therefore, this study aimed to evaluate the effects of different TSA concentrations on histone hyperacetylation during in vitro cell differentiation of murine pluripotent embryonic stem cells, cultured with or without LIF, in the quest of to standardize their application on early cultures of embryonic stem cells.Materials, Methods & Results: Undifferentiated murine embryonic stem cells were plated in the presence of different TSA concentrations (0 nM, 15 nm, 50 nM and 100 nM) in the presence or absence of LIF. Thus, the treatments were evaluated in undifferentiated embryonic stem cells cultured in the presence of LIF (Control group: 0 nM LIF(+); Group 15 nM LIF+; Group 50 nM LIF+ and Group 100 nM LIF+), and in embryonic stem cells cultured in the absence of LIF (Control group: 0 nM LIF; Group 15 nM LIF(-); Group 50 nM LIF(-) and Group 100 nM LIF-). Treatment with TSA was performed for 24 h. After that the medium was replaced with fresh medium without TSA. Samples were collected at 0, 12, 24, 36 and 48 h after the beginning of the experiment. Three replicates were performed in each experimental group. The relative amount of Histone H3 lysine 9 acetylation was analyzed in all groups, as well as the cell proliferation in the embryonic stem cells cultured in the presence of LIF. In the control group (0 nM), the absence of LIF resulted in higher levels (P < 0.05) of H3lys9ac compared to the cultures supplemented with LIF. In the embryonic stem cells cultured in the presence of LIF, the 50 nM and 100 nM treatments resulted in higher levels (P < 0.05) of H3lys9ac when compared with 0 nM and 15 nM treatments. Evaluating the Hoechst area in the 0 nM group, it was observed that the number of cells increased (P < 0.05) according to the time of culture. Treatment with 15 nM also reflected a similar distribution, but the Hoechst area in 15 nM group was lower (P < 0.05) at 24 and 48h when compared to the observed in the control group. In the 100 nM treatment, was observed that the area of Hoechst was lower (P < 0.05) to that obtained in the control group at 12, 24 and 48h. In addition, it was observed that treatment with TSA induces greater cellular differentiation when compared to control groups in stem cells cultured in the presence of LIF as well as in the absence of LIF.Discussion: In the present study it was observed that TSA treatment increased the levels of histone acetylation in murine embryonic stem cells at a 50 nM concentration, making it possible to reduce the concentration recommended in the literature (100 nM). In addtion, it was concluded that the lower TSA concentrations utilized (15 nm and 50 nM) was less harmful to cellular proliferation than the 100 nM TSA concentration.

Isolamento, caracterização e diferenciação de células-tronco mesenquimais do líquido amniótico equino obtido em diferentes idades gestacionais

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

INTERAÇÕES ENTRE MAGNÉSIO E MICRONUTRIENTES METÁLICOS DURANTE O DESENVOLVIMENTO DE FEIJOEIRO, em SOLUÇÃO NUTRITIVA

Este estudo avaliou as possíveis interações entre o magnésio e os micronutrientes manganês, zinco, cobre e ferro, pela determinação de seus teores nos diferentes órgãos do Phaseolus vulgaris L. cv Carioca, nas 5 coletas, realizadas a cada 14 dias, a partir do 25o dia após a semeadura. As plantas foram cultivadas em solução nutritiva com nível completo (48,6 mg.L-1), baixos (2,4 e 24,3 mg.L-1) e elevados (72,9 e 97,2 mg.L-1) de Mg. O delineamento experimental foi inteiramente casualizado, em esquema fatorial. Com os níveis baixos de Mg, observou-se a diminuição do teor desse mineral nas raízes, caule e folhas e de Cu nas folhas e o aumento de Zn nas folhas e de Mn nas raízes. Com 97,2 mg.L-1 de Mg, observou-se o aumento desse elemento nas folhas, sugerindo sua absorção e translocação das raízes para a parte aérea. Nesses feijoeiros também observou-se diminuição de Cu no caule e nos frutos e de Mn nas raízes. As concentrações de Mg e Mn nas raízes variaram sempre de modo oposto, com os diferentes tratamentos utilizados. Isso sugere uma relação inversa na absorção desses nutrientes. Os resultados demonstraram uma pequena influência da variação dos níveis de Mg na solução sobre os micronutrientes que, no entanto, pode alterar o comportamento da planta.

Gibberellin and cytokinin effects on soybean growth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Soluções nutritivas no desenvolvimento do crisântemo cultivado em vaso

Although the information about nutritive solution for potted chrysanthemum is still incongruent, the objective of this work was to evaluate the effect of nutritive solutions in the development of Miramar chrysanthemum cultivated in pots at FCAV-UNESP, Jaboticabal-SP. The experiment had a randomized block design with split-plot evaluation and treatments corresponding to four nutritive solutions consisting of commercial fertilizers (S1, S2, S3, S4) evaluated at 0, 14, 28, 42, 56 and 70 days after rooting (DAE), with five replications. The plant height, leaf area, number of leaves, stem diameter, total dry matter mass of aerial part and roots; number and diameter of inflorescences, and physiological indexes of growth analysis were evaluated. The nutritive solutions did not interfere significantly in stem diameter, number and diameter of inflorescences and dry matter of roots, although S3 and S4 provided the highest plants, leaf area, number of leaves, total dry matter of aerial part. The physiological indexes showed that plants presented an accelerated growth from 14 to 28 DAE with S4 and S3 solutions and therefore they were considered the best solutions.

An Educational international partnership responding to local needs: Process evaluation of the Brazil FAIMER Regional Institute

The Brazilian public health system requires competent professionals sensitive to the needs of the population. The Foundation for Advancement of International Medical Education and Research (FAIMER) provides a two-year faculty development programme for health professions educators, aiming to build leadership in education to improve health. A partnership with governmental initiatives and FAIMER was established for meeting these needs. This paper describes the initial process evaluation results of the Brazilian FAIMER Institute Fellowship (FAIMER BR). Methods: Data were analysed for the classes 2007-2010 regarding: application processes; innovation project themes; retrospective post-pre self-ratings of knowledge acquisition; and professional development portfolios. Results: Seventeen of 26 Brazilian states were represented among 98 Fellows, predominantly from public medical schools (75.5%) and schools awarded Ministry of Health grants to align education with public health services (89.8%). One-third (n = 32) of Fellows' innovation projects were related to these grants. Significant increases occurred in all topic subscales on self-report of knowledge acquisition (eff ect sizes, 1.21-2.77). In the follow up questionnaire, 63% of Fellows reported that their projects were incorporated into the curriculum or institutional policies. The majority reported that the programme deepened their knowledge (98%), provided new ideas about medical education (90%) and provided skills for conflict management (63%). One-half of the Fellows reported sustained benefits from the programme listserv and other communications, including breadth of expertise, establishment of research collaboration and receiving emotional support. Conclusion: Contributors to initial programme success included alignment of curriculum with governmental initiatives, curriculum design merging educational technology, leadership and management skills and central role of an innovation educational project responding to local needs.

Crescimento inicial e nutrição de cedro (Cedrela fissilis Vell.) em função de doses de nitrogênio

We aimed to evaluate the effects of nitrogen rates in early growth, SPAD index, content and accumulation of nutrients in cedar seedlings. The experiment was carried out in a plastic greenhouse and the seedlings were grown in plastic pots filled with 20 dm-3of Rhodic Hapludox, arranged inrandomized blocks with four replications. Rates of 0, 40, 80, 120 and 160 mg dm-3N were tested. Fertilization up to 160 mg dm-3N promoted increases in SPAD index and early growth of plants, beyond greater absorption of N,P, K, Ca, Mg, S, B, Cu, Fe, Mn and Zn. Overall, lower N rates limited growth but not the absorption of nutrients by cedar plants. In leaves, the only nutrien taffected was the B which their content was reduced with increasing of N rate. Increased of nitrogen fertilization changed the Cu distributionin plants, since increased Cu content in roots and reduced their content in stem. Higher N rates promoted higher accumulation of all nutrients due to the increase of dry matter. Nitrogen fertilization changed the dynamic of nutrient absorption in cedar. With 160 mg dm-3N, the amount absorbed followed the order: N>S>Ca>K>Mg>P>Fe>Mn>B> Zn> Cu.