A histone deacetylase inhibitor, azelaic bishydroxamic acid, shows cytotoxicity on Epstein-Barr virus-transformed B-cell lines - A potential therapy for posttransplant lymphoproliferative disease

Background. Posttransplant lymphoproliferative disease (PTLD), driven by the presence of Epstein-Barr virus (EBV), is becoming an increasingly important clinical problem after solid organ transplantation. The use of immunosuppressive therapy leads to the inhibition of the cytotoxic T cells that normally control the EBV latently infected B cells. The prognosis for many patients with PTLD is poor, and the optimal treatment strategy is not well defined. Method. This study investigates the use of a histone deacetylase inhibitor, azelaic bishydroxamic acid (ABRA), for its ability to effectively kill EBV-transformed lymphoblastoid cell lines. Results. In vitro treatment of lymphoblastoid cell lines with ABRA showed that they were effectively killed by low doses of the drug (ID50 2-5 mug/ml) within 48 hr. As well as being effective against polyclonal B-cell lines, ABHA was also shown to be toxic to seven of eight clonal Burkitt's lymphoma cell lines, indicating that the drug may also be useful in the treatment of late-occurring clonal PTLD. In addition, ABHA treatment did not induce EBV replication or affect EBV latent gene expression. Conclusion. These studies suggest that ABHA effectively kills both polyclonal and clonal B-cell lines and has potential in the treatment of PTLD.

Cultivo in vitro de Crambe abyssinica Hochst. : germinação, micropropagação, estabilidade genética e anatomia foliar

Crambe (Crambe abyssinica) pertence à família Brassicaceae, originário da Etiópia e principalmente destinado à produção de forragem (30 a 32% de proteína bruta). Atualmente, tem sido bastante cultivado visando à extração de óleo vegetal. Com os atuais incentivos à busca de fontes de energias renováveis, o cultivo de crambe vem ganhando papel de destaque na produção de biodiesel por suas diversas vantagens, como: (a) rápido ciclo de vida (colhida em torno de 90 dias); (b) alta produção de biomassa; (c) alta produtividade de sementes (1000 e 1500 kg ha-1); (d) menor custo de produção em relação a outras fontes oleaginosas, como, canola, girassol e soja; (e) um percentual de óleo total na semente entre 32 e 38%, superando, por exemplo, a soja; (f) potencial de fitorremediação, eficiente na descontaminação de arsênio, cromo e outros metais pesados; e (g) elevado percentual de ácido erúcico (50 a 60%) sendo útil na indústria de plástico e lubrificante. Devido aos poucos trabalhos realizados com crambe, abre-se um vasto campo de investigações científicas que tenham como objetivo desenvolver as potencialidades dessa cultura e, consequentemente, melhorar os aspectos agronômicos e tecnológicos para seu emprego na indústria de biodiesel. Nesse contexto, as técnicas de cultivo in vitro foram importantes tanto para a propagação massal, quanto como ferramenta para uma possível aplicação de outras técnicas biotecnológicas, contribuindo para uma produção homogênea, fiel e em larga escala. Portanto, este trabalho teve como objetivo geral avaliar as condições mais favoráveis à germinação, estabelecimento in vitro e micropropagação de Crambe abyssinica Hochst., além de verificar possíveis alterações genéticas e anatômicas, possibilitando a regeneração e produção de plântulas viáveis. Para a germinação e estabecimento in vitro de crambe, as condições mais favoráveis foram em meio B5 ou WPM, na presença ou ausência de pericarpo e na presença de luz. Na micropropagação dessa espécie, uma frequência satisfatória de regeneração de brotos foi obtida a partir de segmentos apicais utilizados como explante em meio contendo 5 μM de BAP (6- benzilaminopurina), e o alongamento foi satisfatório com 1 μM de GA3 (ácido giberélico). Os marcadores moleculares ISSR (Inter-Simples Sequence Repeats) utilizados para a análise da estabilidade genética indicaram que o segmento apical de crambe é um explante confiável para a micropropagação de plantas geneticamente verdadeiras (true-to-tipe), ou seja, mantém a estabilidade genética. As diversas fontes de citocininas e concentrações utilizadas neste trabalho não promoveram mudanças, no sentido de alterar a organização e/ou a espessura em relação ao controle, e as alterações observadas na estrutura e espessura das folhas dos tratamentos de aclimatização prejudicaram o processo de estabelecimento da plântula ex vitro. Contudo, existe a necessidade de um enraizamento e aclimatização eficiente para completa propagação in vitro de crambe. Portanto, este protocolo de regeneração de plantas in vitro de crambe pode ser útil no processo de criação e desenvolvimento de novas cultivares em um tempo mais curto e no melhoramento genético usando explantes apicais.

Gladiolus development in response to bulb treatment with different concentrations of humic acids

Gladiolus is an ornamental species produced for cut flowers and propagated by corms. The early flowering and increase in the number of flower buds, besides the production of commercial corms are constant challenges to be addressed in the crop improvement. Commercial production of ornamentals is technologically accelerated by means of growth regulators. Among them, the auxins stand out for their key role in the adventitious rooting and cell elongation. Alternatively, the humic substances present in the organic matter also have biostimulating effect, which is very similar to the auxinic effect. Therefore, this work aimed to study the growth and development of gladiolus in response to application of different concentrations of humic acids (HA) isolated from vermicompost. Corms were soaked for 24 hours in solutions containing 0, 10, 20, 30 and 40 mmol L-1 of C from HA. The corms were planted in 10-dm³ plastic bags filled with substrate and kept in a greenhouse. Growth of shoots and roots was evaluated. The results showed that the use of HA accelerates growth, and anticipates and increases flowering of Gladiolus.

Isoenzyme profiles of Trypanosoma cruzi stocks from different areas of Paraguay

Twenty one Trypanosoma cruzi stocks from humans, domiciliary triatomines and one sylvatic animal of different areas of Paraguay were subjected to isoenzyme analysis. Thirteen enzyme systems (15 loci in total) were studied. MN cl2 (clonets 39) and SO34 cl4 (clonets 20) were used as references. Relationships between stocks were depicted by an UPGMA dendrogram constructed using the Jaccard´s distances matrix. Among the Paraguayan stocks 14 zymodemes were identified (Par1 to Par14), Par 5 being the most frequent. Polymorphism rate and clonal diversity were 0.73 and 0.93, respectively. Average number of alleles per polymorphic locus was 2.5 (range 2-4). These measurements show a high diversity, which is confirmed by the dendrogram topology. All stocks belong to the same lineage, as MN cl2 reference strain (T. cruzi II). Moreover three distinct subgroups were identified and two of them correspond to Brazilian and Bolivian zymodemes, respectively. The third subgroup, the most common in Paraguay, is related to Tulahuen stock. The large geographical distribution of some zymodemes agrees with the hypothesis of clonality for T. cruzi populations. However sample size was not adequate to detect genetic recombination in any single locality.

Proliferation is a prerequisite for bacterial superantigen-induced T cell apoptosis in vivo.

Staphylococcal enterotoxin B (SEB) is a bacterial superantigen that binds to major histocompatibility complex class II molecules and selectively interacts with T cells that bear certain T cell receptor (TCR) V beta domains. Administration of SEB in adult mice results in initial proliferation of V beta 8+ T cells followed by a state of unresponsiveness resulting from a combination of clonal deletion and clonal anergy in the SEB-reactive population. At this time, it is unclear what relationship exists between the T cells that have proliferated and those that have been deleted or have become anergic. Here we show that only a fraction of the potentially reactive V beta 8+ T cells proliferate in response to SEB in vivo, and that all the cells that have proliferated eventually undergo apoptosis. Virtually no apoptosis can be detected in the nonproliferating V beta 8+ T cells. These data demonstrate a causal relationship between proliferation and apoptosis in response to SEB in vivo, and they further indicate that T cells bearing the same TCR V beta segment can respond differently to the same superantigen. The implications of this differential responsiveness in terms of activation and tolerance are discussed.

Model development for capercaillie (Tetrao urogallus) habitat in the Jura Mountains (Western Switzerland)

Capercaillie, Tetrao urogallus, is a threatened species in central Europe, and Swiss populations declined 40 to 50 % between 1970 and 1985. Capercaillie are sensitive to forest structure, and loss of habitat is a major cause of their decline. Knowledge of habitat characteristics is therefore essential for capercaillie conservation. Here, we present models predicting capercaillie probability of occurrence, based on relevant structural habitat variables. Models were built using multiple logistic regression analyses on capercaillie presence/absence data. Vegetation survey was carried out in July 1999 in a 170-km2 forested area (Jura mountains, canton de Vaud, western Switzerland) inhabited by capercaillie and presence/absence of the species was assessed according to dropping presence/absence. The survey was based on 10-m-radius sample plots each in a 1-km2 forest patch (n = 76 with capercaillie droppings, n = 80 without). A first model included seven out of 27 measured habitat variables and a second model only four. The latter model best represents practical needs. It includes three variables which had a negative impact on capercaillie presence: tree and shrub covers and spruce, Picea excelsa, shrub cover, and one which had a positive effect: bilberry, Vaccinium myrtillus, cover, highlighting that capaercaillie selected open forest with high bilberry abundance. The model can be used to map potential capercaillie habitat distribution and to manage the habitat in favour of capercaillie (protection and adapted forestry practices) in the Swiss Jura mountains.

THE LYCHEE TREE PROPAGATION BY LAYERING

The aim of the study was to assess the influence of season and different substrates on rooting of air layers of lychee (Litchi chinensisSonn.) for the production of seedlings to ensure the formation of uniform and productive orchards. Air layers were done in plants of the Bengal cultivar using leafy and healthy woody branches, with about 0.010 to 0.015 m in diameter, in which were performed complete girdling with 0.020 m wide at a distance of 0.30 to 0.40 m below the apex. Then the branches were wrapped in moistened substrate. The layering was made at six times of theyear (January, March, May, July, September and November) and two substrates were used (coconut fiber and sphagnum) in a 6 x 2 factorial design in a randomized block with ten replicates. After 90 days, layers were separated from the matrix plant and evaluated for rooting and callus formation, root number, considering only the primary roots, length, area and volume of the roots, beyond the dry weight of roots and calluses. The months of January, March, September and November showed the best results for all analyzed variables related to rooting. With respect to the substrates, the only difference was in January and March to the root number and dry weight of roots, where the sphagnum showed the best results. The month of July was more conducive to the formation of calluses. The period between September and March was more suitable to the propagation of lychee, when there were rooting percentages above 90%, in addition to the formation of large amount of roots.

Metsäsääntely Suomessa ja Venäjällä. Näkökulmia kestävään metsätalouteen

Sustainability is the aim of forest management and forest regulation in many countries. Accordingly, forest management has been steered towards more environmentally friendly methods and new regulatory instruments have been introduced. At the same time, wood trade and forest industry have become a global business. Even if the importance of national forest legislation has not decreased, it has been widely acknowledged that national regulation of forest management is no longer sufficient. The movement of goods does not acknowledge boundaries, even though most negative environmental and social consequences stay in the country of origin of wood and other raw materials. As a partial solution to this dilemma, different kinds of regulations have been developed. Various forest certification schemes and wood trade regulation in the EU (995/2010) are examples of efforts to prevent illegal logging and unsustainable forestry. The Finland-based forest industry is to a varying extent dependent on wood trade from Russia. Especially in the 1990‟s, ethical questions concerning import of wood from Russian old growth forests near the Finnish border were widely discussed. Consequently, forest industry enterprises have developed systems to trace the origin of wood and to buy certified wood from Russia. The aim of the research has been to evaluate Finnish and Russian forest regulations in order to investigate what kind of forest management these regulations enhance, and to what extent ecologically sustainable forest management has been integrated into different forms of regulation. I have examined Finnish and Russian forest regulation in four separate articles based on the topics of the Russian Forest Code, forest certification and other voluntary forest protection measures in Russia, Finnish forest certification and Finnish forest legislation. One objective has been to analyse the roles voluntary forest certification plays in promoting sustainable forest management in different countries. In my research, I have mainly concentrated on ecological sustainability and protection of biodiversity, although other aspects of sustainable forest management have been touched upon in different articles. In the following I shall conclude the findings of my research. When the current Russian Forest Code (2006) was being adopted, the main emphasis was not on ecological issues, but on reorganizing forest governance. The role of ecological requirements was even slightly diminished during the legislative reform. There are, nevertheless, still stipulations aiming at ecological sustainability, such as the division of forests into different forest management categories and various protection zones. In 2000, FSC forest certification arrived in Russia, at present covering already 28 million hectares of forests. The PEFC scheme is now in use as well, but to a much lesser extent. If properly implemented, Russian forest certification schemes clearly improve the level of ecological and social sustainability of forestry in Russia. Certification criteria, however, are partly in conflict with the Russian forest legislation and certified enterprises have been forced to pay fines or to negotiate with forest authorities. This clearly indicates that even if Russian forest legislation has otherwise been liberalized to a certain extent, some significant paternalism still exists. Voluntary, hands-on biodiversity protection measures are not valued, and they are not part of the official protection policies as in many other countries. However, there have been some regional solutions to this dilemma. In the Republic of Karelia forest authorities have approved a set of forest biodiversity protection rules created by a local NGO and a forest industry enterprise. By following these local rules, an enterprise can avoid fines for protection measures. The current Finnish Forest Act was adopted in 1996. It brought forest legislation into a new era as some ecological aspects were integrated into forest legislation. The various soft-law forest management recommendations further increased the level of biodiversity protection. My evaluation of the overall legitimacy of the Finnish forest legislation and forest management paradigm revealed, however, several problematic issues. As part of this study I analysed the history of the current forest management paradigm. This analysis revealed the path dependency which still hinders the protection of biodiversity and clearly decreases the general legitimacy of forest management. Due to several historical reasons only even-structured forest management based on clear cuts has for decades been officially approved in Finland. Due to increasing demands of forest owners the legislation is finally being revised. Yet, the official approval of uneven-structured forest management would not be enough to fully improve ecological, social and cultural legitimacy. The latest ecological theories and knowledge of endangered species should be taken into account in the on-going reform of forest legislation as well as the modernisation. Forest legislation is one of the very few spheres of Finnish environmental legislation where openness and participation are still considered a threat. The first Finnish forest certification scheme, PEFC, was established in 2000. It now covers more than 20 million hectares, about 95% of the forests in Finland. PEFC Finland does not require a higher level of biodiversity protection than the recommendations by Tapio (the Development Centre for Forestry), but certification has unified forest management practices and requires more protection measures than mere forest legislation. The study suggests that in Finland PEFC has not functioned as an instrument which would substantially improve the level of forest management. Rather it has supported the status quo of the forest sector. While the ecological and social responsibility of Finland-based forest corporations was one impetus for this research, I want to conclude that there are problems related to forest legislation and non-state regulation in both Finland and Russia. If an enterprise buying wood from Russia buys only certified wood, and carefully avoids wood coming from high conservation value forests that are either ecologically or socially valuable, it can be claimed to be as sustainably produced as in Finland. However, there must be continuous scrutiny of the circumstances. In Russia, the level of the compliance of certification criteria varies considerably, and there are vast unprotected invaluable forest areas. The utilisation of these areas should not be based on short-sighted decisions or lack of consensus among stakeholders.

Mixed reproduction systems in Opuntia monacantha (Cactaceae) in Southern Brazil

The Cactaceae have morphological and physiological adaptations associated with their life histories that are reflected in different modes of reproduction and multiplication. The reproductive phenology, preferential reproductive mode, and the sexual and asexual multiplication of Opuntia monacantha were investigated between 2003 and 2006 in two restinga (sandy coastal) environment in Southern Brazil to determine the adaptive values of its reproduction modes. Flowering was annual and occurred continuous for approximately 100 days between the months of September and January, with a few flowers opening per day per individual (but many in the whole population). Facultative xenogamy was identified as the preferential sexual system, with the highest levels of fruit and seed formation following natural pollination. The seeds are recalcitrant and have high germinative capacities under laboratory conditions. Seedling recruitment and establishment in the field was extremely reduced. The skins of the fruits and the cladodes have high capacities for regeneration and clonal multiplication because of their areolas.

Conventional cytogenetic characterization of a new cell line, ACP01, established from a primary human gastric tumor

Gastric cancer is the second most frequent type of neoplasia and also the second most important cause of death in the world. Virtually all the established cell lines of gastric neoplasia were developed in Asian countries, and western countries have contributed very little to this area. In the present study we describe the establishment of the cell line ACP01 and characterize it cytogenetically by means of in vitro immortalization. Cells were transformed from an intestinal-type gastric adenocarcinoma (T4N2M0) originating from a 48-year-old male patient. This is the first gastric adenocarcinoma cell line established in Brazil. The most powerful application of the cell line ACP01 is in the assessment of cytotoxicity. Solid tumor cell lines from different origins have been treated with several conventional and investigational anticancer drugs. The ACP01 cell line is triploid, grows as a single, non-organized layer, similar to fibroblasts, with focus formation, heterogeneous division, and a cell cycle of approximately 40 h. Chromosome 8 trisomy, present in 60% of the cells, was the most frequent cytogenetic alteration. These data lead us to propose a multifactorial triggering of gastric cancer which evolves over multiple stages involving progressive genetic changes and clonal expansion.

Forest inventory information needs in Northwest Russia

This thesis applies the customer value hierarchy model to forestry in order to determine strategic options to enhance the value of LiDAR technology in Russian forestry. The study is conducted as a qualitative case study with semi-structured interviews as a main source of the primary data. The customer value hierarchy model constitutes a theoretical base for the research. Secondary data incorporates information on forest resource management, LiDAR technology and Russian forestry. The model is operationalised using forestry literature and forms a basis for analyses of primary data. Analyses of primary data coupled with comprehension of Russian forest inventory system and knowledge on global forest inventory have led to conclusions on the forest inventory methods selection criteria and the organizations that would benefit the most from LiDAR technology use. The thesis recommends strategic options for LiDAR technology’s value enhancement in Russian forestry.

Forest inventory information needs in Northwest Russia

This research report applies the customer value hierarchy model to forestry in order to determine strategic options to enhance the value of LiDAR technology in Russian forestry. The study is conducted as a qualitative case study with semi-structured interviews as a main source of the primary data. The customer value hierarchy model constitutes a theoretical base for the research. Secondary data incorporates information on forest resource management, LiDAR technology and Russian forestry. The model is operationalised using forestry literature and forms a basis for analyses of primary data. Analyses of primary data coupled with comprehension of Russian forest inventory system and knowledge on global forest inventory have led to conclusions on the forest inventory methods selection criteria and the organizations that would benefit the most from LiDAR technology use. The report recommends strategic options for LiDAR technology’s value enhancement in Russian forestry. This work has been conducted as a part of the project ‘Finnish-Russian Forest Academy 2 - Exploiting and Piloting’, which has been supported financially by the South-East Finland- Russia ENPI CBC 2007-2014 Programme.

Influência das principais commodities agropecuárias e de variáveis macroeconômicas sobre os preços da terra agrícola no Brasil

Este trabalho tem como proposta investigar como o preço de terras de uso rural no Brasil é afetado pelos preços e exportações das principais commodities agropecuárias, bem como por variáveis macroeconômicas, como taxa básica de juros, taxa de câmbio, taxa de inflação e disponibilidade de crédito agrícola. Para tal foram consideradas as produções agrícola de algodão, café, cana-de-açúcar (e seus principais produtos açúcar e etanol), milho e soja, a produção pecuária de carne bovina e a produção industrial de celulose de fibra curta com foco em sua principal matéria prima, os plantios reflorestados de eucalipto. Em linha com estudos anteriores, foi encontrada evidência empírica de que o preço da terra possui cointegração com algumas das variáveis agrícolas, pecuárias e florestais citadas, em especial em estados com maior vocação agropecuária e/ou para silvicultura. Quanto às variáveis macroeconômicas, apenas a taxa básica de juros apresentou cointegração com o preço de terras para todos os estados avaliados, taxa de câmbio e disponibilidade de crédito rural não aparecem como variáveis estatisticamente significantes. Conclui-se que, para estados com notável participação na balança comercial brasileira de produtos agrossilvipastoris, é possível obter um modelo de equilibro de longo prazo entre o preço da terra de uso rural e as variáveis destacadas acima, de modo que investidores do setor possam utilizá-lo como ferramenta de projeção no auxílio da tomada de decisão além de avaliar potenciais impactos no valor de seus ativos A inovação do presente estudo está em testar as hipóteses de cointegração para cada um dos estados da federação.

Emprego da citometria de fluxo na avaliação do perfil imunofenotípico de pacientes com leucemia linfocítica crônica

Chronic lymphocytic leukemia (B-CLL) is a clonal proliferation of mature B lymphocytes characterized by indolent clinical course. Biologically this clonallity is characterized by low expression of surface immunoglobulin (sIg) with restriction to a single immunoglobulin light chain associated with high expression of CD5 antigen and positivity to B cell antigens lymphocytes such as CD19, CD20 and CD23 and negativity to FMC7. The immunological profile and morphological analysis of lymphoid cells are the main means for the differential diagnosis of B-CLL from other chronic lymphoproliferative diseases. The aim of this study was to evaluate the expression pattern of a variety of membrane antigens in leukemic cells originating from patients with B-CLL. In this study, peripheral blood samples from 80 patients with B-CLL were analyzed by multiparametric flow cytometry in addition to routine hematologic exams, using a panel of monoclonal antibodies (MoAb): CD45/CD14, CD3/CD19/CD45, CD4/CD8 / CD3, CD20/CD5/CD3, CD3/CD16-56/CD45, CD2/CD7, FMC7/CD23, CD103/CD22/CD20, HLADR/CD38, CD10/CD19, CD1a, CD11b and also IgM/gD, kappa and lambda immunoglobulin light chains for the detection of surface immunoglobulin and clonal restriction for immunoglobulin light chain. The Hematological data were obtained from the hematological analyzer and cytomorphological analysis in blood film stained by Leishmann. The study samples consisted of 45 men and 35 women, ages ranging from 55 to 84 years (mean 65 years). Complete white blood count showed count ranging from 10.0 to 42.0 x 109/l. (mean 50.0 x 109/l) and lymphocytes count greater than 5.0 x 109/l in all cases. The neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/CD20+/HLADR+/CD23+) in the vast majority of the cases, associated to failed to stain for T cell markers (CD1a, CD2, CD4, CD3, CD7, CD8), CD103, CD14 and FMC7. Leukemic cells of most patients also expressed low intensity of IgM and IgD with restricted kappa light chain, in most cases (59,7%). This observation highlights the importance of immunophenotyping for correct diagnosis of chronic lymphoproliferative syndromes and the panel of MoAb used was sufficient for diagnostic confirmation of B-CLL

Efeito do comprimento de estacas herbáceas de dois clones de umezeiro (Prunus mume Sieb & Zucc.) no enraizamento adventício

O umezeiro (Prunus mumeSieb & Zucc.) é uma rosácea de folhas caducas, nativa da China, cujos frutos e flores são muito apreciados pelos povos orientais. No Brasil, alguns estudos foram realizados visando a sua utilização como porta-enxerto para pessegueiro e nectarineira, dadas as suas características de adaptação, rusticidade, redução do porte da planta e compatibilidade com algumas cultivares de Prunus persica. O presente estudo foi conduzido em câmara de nebulização sob ripado, pertencente ao Departamento de Produção Vegetal da FCAV/UNESP, Câmpus de Jaboticabal-SP. Objetivou-se verificar a influência de quatro comprimentos de estacas herbáceas no enraizamento de dois clones de umezeiro. O material vegetal, identificado como Clone 10 e Clone 15, foi oriundo do Programa de Melhoramento Genético do Instituto Agronômico de Campinas-SP. O experimento foi constituido de fatorial 2 x 4, em blocos casualizados, sendo o fator clone em 2 níveis (Clone 10 e Clone 15) e o fator comprimento de estaca em 4 níveis (12; 15; 18 e 25cm). Pelos resultados observados, verificou-se diferença entre os clones somente na porcentagem de estacas brotadas e número de raízes por estaca. O comprimento da estaca influenciou na porcentagem de enraizamento e na mortalidade das estacas, sendo que estacas maiores tenderam a apresentar maiores porcentagens de enraizamento e menores de mortalidade. As estacas com 12cm, embora apresentando menor número de raízes por estaca, são recomendadas por permitirem a obtenção de um maior número de estacas por planta-matriz. Houve efeito significativo da interação entre os fatores para número e comprimento de raízes.