Estaquia caulinar de guaçatonga (Casearia sylvestris Swartz) nas quatro estações do ano, com aplicação de diferentes concentrações de AIB / Stem cuttings of "guaçatonga" (Casearia sylvestris Swartz) in the four seasons of the year with the use of different IBA concentrations

Casearia sylvestris Swartz (Salicaceae) ou guaçatonga é uma árvore nativa do México, da América Central, e da América do Sul, com grande importância ecológica, farmacológica, e comercial. No entanto, como a maioria das espécies nativas de interesse medicinal no Brasil, a guaçatonga não é cultivada comercialmente, sendo obtida por extrativismo. O presente trabalho foi conduzido com o objetivo de testar um protocolo de propagação vegetativa de guaçatonga por meio da estaquia, visando identificar qual a melhor estação do ano para o enraizamento de estacas e avaliar o efeito da utilização do regulador vegetal ácido indolbutírico (AIB). No outono, inverno e primavera de 2007, e no verão de 2008, estacas caulinares semilenhosas de 12-14 cm de comprimento e com duas folhas foram preparadas e tratadas com AIB (0, 1000, 2000 e 3000 mg L-1), em solução alcoólica, através da imersão rápida por 10 segundos da base das estacas, e foram plantadas em tubetes contendo substrato Plantmax HT® em casa-de-vegetação sob nebulização intermitente. O delineamento experimental utilizado foi o inteiramente casualizado, com 4 repetições, 4 tratamentos, e 16 estacas por parcela. Todos os experimentos foram avaliados após 90 dias, sendo que para dois deles (primavera 2007 e verão 2008) prolongou-se o tempo de permanência em casa-de-vegetação para melhor desenvolvimento das raízes. Foram avaliados os parâmetros: porcentagem de estacas enraizadas, porcentagem de estacas vivas (com calos e sem raízes, sem calos e sem raízes), porcentagem de folhas retidas, porcentagem de estacas mortas, número de raízes, comprimento das três maiores raízes (cm), e média da massa seca das raízes (mg). Não ocorreu enraizamento nas estacas retiradas no outono e no inverno. Com as estacas retiradas na primavera obteve-se 39,1% de enraizamento. Estacas coletadas no verão não responderam como o esperado, apresentando, após 240 dias, 6,3% de enraizamento no tratamento com 3000 mg L-1 de AIB. O AIB até 3000 mg L-1 não estimulou o enraizamento de estacas de guaçatonga e a melhor estação do ano para a estaquia é a primavera.

Casearia sylvestris Swartz (Salicaceae), or "guaçatonga", is a tree native to Mexico and Central and South America, with great ecological, pharmacological and commercial relevance. Similarly to most native species of medicinal interest in Brazil, "guaçatonga" is not commercially cultivated and is obtained by means of extraction. The aim of this study was to test a protocol for the vegetative propagation of "guaçatonga" by means of stem cutting, identifying the best season for stem rooting and assessing the effect of using the growth regulator indolebutyric acid (IBA). In the fall, winter and spring 2007 and summer 2008, semi-hardwood cuttings with 12-14 cm length and two leaves were prepared and treated with IBA (0, 1000, 2000 and 3000 mg L-1), in alcohol solution, by rapidly immersing for 10 seconds the base of cuttings and planting them in tubes containing Plantmax HT® in greenhouse under intermittent nebulization. Experimental design was completely randomized with 4 replicates, 4 treatments and 16 cuttings per plot. All experiments were evaluated after 90 days, and for two of them the time of maintenance in greenhouse was prolonged in order to improve root development. The following parameters were evaluated: percentage of rooted cuttings, percentage of live cuttings (with callus and without roots, without callus and without roots), percentage of retained leaves, percentage of dead cuttings, number of roots, length of the largest roots (cm) and mean dry mass of roots (mg). There was no rooting on cuttings collected during the fall and the winter. For cuttings collected in the spring, 39.1% rooting was obtained. Cuttings collected in the summer did not show the expected results, presenting after 240 days 6.3% rooting when treated with 3000 mg L-1 IBA. Up to 3000 mg L-1, IBA did not stimulate the rooting of "guaçatonga" cuttings and the best season for cutting is spring.

Capacidade de enraizamento de estacas de Maytenus muelleri Schwacke com a aplicação de ácido indol butírico relacionada aos aspectos anatômicos / Rooting capacity of Maytenus muelleri Schwacke cuttings with indolebutyric acid application related to anatomical aspects

A espinheira-santa (Maytenus muelleri - Celastraceae) é a planta medicinal nativa do Sul do Brasil, cujas folhas são tradicionalmente utilizadas pela medicina popular para o tratamento de úlceras e outros problemas gástricos. Existem poucos trabalhos publicados sobre a produção de mudas e técnicas de propagação vegetativa da espécie. A propagação de espinheira-santa por estaquia poderia ser um método eficiente para obtenção de material homogêneo, com características genéticas desejáveis, produzido a partir de plantas matrizes selecionadas. O presente trabalho teve por objetivo estudar os efeitos da aplicação de ácido indol butírico (AIB), em solução e em pó, no enraizamento de estacas de espinheira-santa coletadas nas quatro estações do ano (abril/2005 a janeiro/2006), bem como averiguar, por meio de análises anatômicas e histoquímicas das estacas, a presença de possíveis impedimentos à iniciação do enraizamento adventício. Estacas provenientes de ramos de plantas matrizes de seis anos cultivadas da Estação Experimental do Canguiri, Pinhais, PR, foram coletadas e tratadas com AIB (0, 1500, 3000 mg L-1 ou mg kg-1), em solução alcoólica (50 por cento v/v) e em talco. Aos 365 dias foram avaliadas as porcentagens de estacas enraizadas e mortas, número e comprimento médio de raízes formadas por estaca. Análises anatômicas e histoquímicas com lugol e cloreto férrico foram realizadas. A estação mais promissora para o enraizamento foi o verão/2006 com 62,50 por cento para o tratamento controle, devido à menor lignficação dos ramos no período de intenso crescimento vegetativo. O número médio de raízes formadas por estaca foi de 6,94 (solução) e o comprimento médio de raízes formadas/estaca chegou a 4,82 cm nesta mesma estação. As concentrações de AIB aplicadas não foram eficientes na indução radicial, independentemente do modo de aplicação. Foi detectada a presença de uma camada quase contínua de fibras e braquiesclereídes, a qual constitui barreira anatômica à indução radicial. Os testes histoquímicos revelaram a presença de amido e de compostos fenólicos nas estacas, em todas as estações do ano. A dificuldade ou demora no enraizamento não pode ser justificada pela falta de reservas de amido nos tecidos das estacas, mas pode ser justificada pela presença de compostos fenólicos, possivelmente do grupo dos monofenóis, que causam a degradação do AIA, interferindo negativamente na indução do enraizamento.

"Espinheira-santa" (Maytenus muelleri - Celastraceae) is a medicinal plant native to Southern Brazil, the leaves of which are traditionally used in popular medicine for the treatment of stomach ulcers and other gastric problems. There are few published studies about seedling production and vegetative propagation techniques for this species. The propagation of "espinheira-santa" by cuttings could be an efficient method to obtain homogeneous material, with desirable genetic characteristics, produced from selected mother plants. This paper aimed to study the effects of indolebutyric acid (IBA) application, in solution and in powder, on the rooting of "espinheira-santa" cuttings, collected in four seasons (April/2005 to January/2006), as well as to investigate, by means of cutting anatomical and histochemical analyses, the presence of possible impediments to adventitious rooting initiation. Cuttings from branches of six-year mother plants grown at "Estação Experimental do Canguiri", Pinhais, Paraná State, Brazil, were collected and treated with IBA (0, 1500, 3000 mg L-1 or mg kg-1) in alcoholic solution (50 percent v/v) and in powder. After 365 days, the percentages of rooted and dead cuttings, the number and mean length of roots/cutting were evaluated. Anatomical and histochemical analyses were performed with lugol and ferric chloride. The most promising season for rooting was Summer/2006, with 62.50 percent of rooting for the control treatment, due to the lesser lignification degree of branches in intense vegetative growth period. The mean number of roots/cutting was 6.94 (solution) and the mean length of roots/cutting was 4.82 cm in that same season. The applied IBA concentrations were not efficient in inducing root growth, regardless of the application method. An almost continuous layer of fibers and stone cells was detected, constituting an anatomical barrier for rooting induction. The histochemical tests revealed the presence of starch and phenolic compounds in cuttings, in all seasons. The difficulty or delay in rooting cannot be justified by the absence of starch reserve in the cutting tissues but by the presence of phenolic compounds, possibly of the group of monophenols, which cause IAA degradation, negatively affecting rooting induction.

Dormência e germinação de sementes de Melaleuca alternifolia Cheel / Dormancy and germination of Melaleuca alternifolia Cheel. seeds

A Melaleuca alternifolia Cheel. é uma espécie aromática de expressivo interesse econômico, em função da presença de óleo essencial armazenado no tecido foliar. As sementes, entretanto, apresentam baixo poder germinativo, o que tem dificultado a obtenção de novos materiais genéticos e o avanço tecnológico para produção no Brasil. O presente trabalho teve por objetivo determinar uma metodologia adequada para avaliar o poder germinativo de sementes de melaleuca. Para tanto, foram realizados dois experimentos, um em condições de laboratório e outro em casa-de-vegetação, avaliando-se diferentes tratamentos para promover a germinação da semente. O delineamento experimental empregado foi o inteiramente casualizado, com quatro repetições, sendo nove tratamentos em laboratório e sete em casa de vegetação; a comparação de médias foi realizada pelo teste Tukey, a 5 por cento de probabilidade. Diante dos resultados obtidos, concluiu-se que a pré-embebição das sementes por 18h, em água, é o procedimento mais adequado, dentre os testados, para avaliar o poder germinativo da semente de melaleuca, sendo que em condições de casa de vegetação, o referido tratamento pode ser usado com os substratos areia ou Plantmax.

Melaleuca alternifolia Cheel. is an aromatic species of high economic interest due to the presence of essential oil in its leaf tissue. However, its seeds have low germinative potential, which has made difficult the obtaining of new genetic materials and the technological advance for production in Brazil. The present work aimed to determine a suitable methodology to evaluate the germinative potential of Narrow-leaved Paperbark seeds. Thus, two experiments were carried out, one under lab conditions and other in greenhouse, in order to evaluate different treatments to stimulate seed germination. Experimental design was completely randomized, with four replicates; nine treatments were used in the lab and other seven in greenhouse. Means were compared by the Tukey's test at 5 percent significance. The obtained results suggest that pre-imbibition of seeds for 18h in water is the most suitable procedure to evaluate the germinative potential of Narrow-leaved Paperbark seeds. Under greenhouse conditions, this treatment can be used with the substrates sand or Plantmax®.

Alpha cell autoantibodies: immunological and metabolic follow-up study.

We studied glucagon responses to OGTT and insulin and arginine stimulation in 12 out of 21 patients who were found positive for alpha cell autoantibodies (ACA) during routine screening procedures for autoimmunity in a group of 4080 individuals. The study was repeated in 8 subjects after an average observation period of 42 months. In both studies glucagon plasma levels were normal and independent of ACA titres, ACA ability to fix complement and ACA ability to cross-react with duodenal alpha cells. The clinical significance of ACA remains to be elucidated.

Complement-fixing adrenal autoantibodies as a marker for predicting onset of idiopathic Addison's disease.

In a prospective investigation of the role of adrenal autoantibodies (AA) in predicting the onset of idiopathic Addison's disease, 9 initially non-addisonian AA-positive autoimmune subjects were followed for 42 months. In 4 of these subjects Addison's disease developed within 1-31 months. A fifth had reduced adrenocortical reserve at the start and at the end of the investigation. An AA capable of fixing the membrane attack complex of complement [(C5-C9) F-AA] was detected before onset of the disease in the sera of the 4 patients in whom Addison's disease developed. (C5-C9) F-AA may be involved in the pathogenesis of idiopathic Addison's disease and may be regarded as a marker for individuals in whom idiopathic adrenal insufficiency is likely to develop.

Islet cell and insulin autoantibodies in organ-specific autoimmune patients. Their behaviour and predictive value for the development of type 1 (insulin-dependent) diabetes mellitus. A 10-year follow-up study.

To evaluate the behaviour and predictive value of islet cell and insulin autoantibodies in patients with organ-specific autoimmune diseases, we followed 21 non-diabetic subjects for a mean period of 84 +/- 27 months. Ten patients were persistently seropositive for complement-fixing islet cell antibodies and high titres of immunoglobulin G islet cell antibodies (greater than or equal to 1:8). The prevalence of persistent insulin autoantibodies in this group was 67%. Seven patients (70%) developed Type 1 (insulin-dependent) diabetes mellitus after a latency period of 2-60 months. The predictive value of complement-fixing islet cell antibodies was 65%, and in the presence of both complement-fixing islet cell and insulin autoantibodies the predictive value rose to 76%. Eleven patients were seronegative for complement-fixing islet cell antibodies and had low immunoglobulin G islet cell antibodies titres (less than 1:8) that were either persistent or transient, or that fluctuated during follow-up. The prevalence of persistent insulin autoantibodies in this group was 45%; only one subject developed Type 1 diabetes. The predictive value of persistent islet cell antibodies (complement-fixing positive/negative) was 54%, and it rose to 70% when both islet cell and insulin autoantibodies were present. Individuals with only insulin autoantibodies or immunoglobulin G islet cell antibodies did not develop diabetes mellitus. A high frequency of HLA-DR3 and/or DR4 was found in patients who developed diabetes mellitus. Thus, the presence of both islet cell and insulin autoantibodies in patients with organ-specific autoimmune disease appears to confer the highest risk of progression toward Type 1 diabetes.

Clinical and subclinical organ-specific autoimmune manifestations in type 1 (insulin-dependent) diabetic patients and their first-degree relatives.

Studying 239 Type 1 (insulin-dependent) diabetic patients and 144 of their first-degree relatives, we found a significant prevalence of autoimmune manifestations in both groups, compared with sex- and age-matched control subjects (p less than 0.001). In particular, in diabetic patients we found a high frequency of autoimmune thyroid disease and idiopathic Addison's disease and also a significant prevalence of thyroid (p less than 0.001), parietal cell (p less than 0.05) and adrenal antibodies (p less than 0.05). In the relatives a high frequency of thyroid disease, thyroid, parietal cell and adrenal antibodies and a significant prevalence of islet cell antibodies (p less than 0.05) were detected. In both groups functional glandular tests and gastric biopsies performed on the basis of autoantibody positivity revealed 13 examples of subclinical hypothyroidism, two cases of reduced adrenocortical reserve and five of atrophic gastritis. Autoantibody screening in diabetic patients and their relatives permitted the early diagnosis of the underlying endocrine disorders.

Thyroid autoantibodies: a good marker for the study of symptomless autoimmune thyroiditis.

In 3737 subjects without clinically thyroid disorders we evaluated the incidence of thyroid microsomal and thyroglobulin antibodies. These autoantibodies were found in 7% of a normal population, in 9% of patients with various non-autoimmune diseases, and in 11-16% of groups who either had or were at risk for autoimmune diseases: patients with IDDM, vitiligo, alopecia areata, idiopathic hypoparathyroidism, Addison's disease, and first-degree relatives of IDDM patients. Functional thyroid evaluation with TRH test was performed in 197 seropositive subjects and 144 seronegative controls. One-quarter (26%) of the subjects with thyroid autoantibodies showed functional abnormalities on TRH testing, whereas only 2.8% of the 144 seronegative controls showed subclinical hypothyroidism. After an observation period of 12-44 months, 102 persistently seropositive subjects were reassessed and 31% of them showed an impairment in TRH test response.