Prostaglandin E2 Prevents Hyperosmolar-Induced Human Mast Cell Activation through Prostanoid Receptors EP2 and EP4.

Background: Mast cells play a critical role in allergic and inflammatory diseases, including exercise-induced bronchoconstriction (EIB) in asthma. The mechanism underlying EIB is probably related to increased airway fluid osmolarity that activates mast cells to the release inflammatory mediators. These mediators then act on bronchial smooth muscle tocause bronchoconstriction. In parallel, protective substances such as prostaglandin E2 (PGE2) are probably also released and could explain the refractory period observed in patients with EIB. Objective: This study aimed to evaluate the protective effect of PGE2 on osmotically activated mast cells, as a model of exercise-induced bronchoconstriction. Methods: We used LAD2, HMC-1, CD34-positive, and human lung mast cell lines. Cells underwent a mannitol challenge, and the effects of PGE2 and prostanoid receptor (EP) antagonists for EP14 were assayed on the activated mast cells. Betahexosaminidase release, protein phosphorylation, and calcium mobilization were assessed. Results: Mannitol both induced mast cell degranulation and activated phosphatidyl inositide 3-kinase and mitogenactivated protein kinase (MAPK) pathways, thereby causing de novo eicosanoid and cytokine synthesis. The addition of PGE2 significantly reduced mannitol-induced degranulation through EP2 and EP4 receptors, as measured by betahexosaminidase release, and consequently calcium influx. Extracellular-signal-regulated kinase 1/2, c-Jun N-terminal kinase,and p38 phosphorylation were diminished when compared with mannitol activation alone. Conclusions: Our data show a protective role for the PGE2 receptors EP2 and EP4 following osmotic changes, through the reduction of human mast cell activity caused by calcium influx impairment and MAP kinase inhibition.

Influência da saponificação na determinação de carotenoides em maracujás do cerrado

This work describes the evaluation of the effect of saponification process in the carotenoid's content of three species of passion fruit. The results indicated the saponification of the extract was necessary to obtain cis-violaxanthin, trans-violaxanthin and β-cryptoxanthin hydrolyzed. These compounds were found in fruits of commercial P. edulis and yellow wild P. edulis. However, the extract saponification did not permitted to obtain free carotenes in fruits of wild purple P. edulis and P. setacea, and to trans-violaxanthin of P. cincinnata, therefore saponification was not indicated in the carotenoid analysis of these three accessions of passion fruit.

Analysis of passion fruit rinds (Passiflora edulis): isoorientin quantification by HPTLC and evaluation of antioxidant (radical scavenging) capacity

The content of isoorientin in passion fruit rinds (Passiflora edulis fo. flavicarpa O. Degener) was determined by HPTLC (high performance thin layer chromatography) with densitometric analysis. The results revealed a higher amount of isoorientin in healthy rinds of P. edulis (92.275 ± 0.610 mg L-1) than in rinds with typical symptoms of PWV (Passion fruit Woodiness Virus) infection (28.931 ± 0.346 mg L-1). The HPTLC data, allied to assays of radical scavenging activity, suggest the potential of P. edulis rinds as a natural source of flavonoids or as a possible functional food.

Histopathological features of infections caused by Fusarium oxysporum and F. solani in purple passionfruit plants (Passiflora edulis Sims)

The purple passionfruit plant, Passifloraedulis Sims, ranks second in fruit exportation in Colombia, and its main destination is the European market. However, its production is affected by several diseases, including fusariosis. This paper presents the histopathological features of different tissues affected by the pathogens Fusarium oxysporum and Fusarium solani. Both microorganisms produce similar responses on the plant: colonization of xylem vessels by hyphae and microconidia, hypertrophy and hyperplasia of the cambium, xylem and phloem; destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant. However, there are differences in the colonization mechanism, F. solani penetrates and is concentrated especially at the collar zone, while F. oxysporum penetrates the roots and moves through the vascular system to colonize the plant.

Intoxicação experimental por Passiflora foetida (Passifloraceae) em caprinos

Passiflora foetida é mencionada por produtores da região Nordeste como causa de intoxicação espontânea em animais. Este trabalho teve por objetivos avaliar a toxicidade de P. foetida em caprinos e determinar seu princípio ativo e a época do ano em que a mesma é tóxica. Inicialmente a planta administrada em duas doses diárias de 40g por kg de peso animal (g/kg) coletada dois dias antes da administração não resultou tóxica. Posteriormente a planta administrada imediatamente após a coleta resultou tóxica nas doses que variaram de 4 a 8 g/kg, em quatro caprinos. O animal que recebeu 8g/kg apresentou sinais clínicos graves e recuperou-se após a administração de tiossulfato de sódio. Os demais caprinos apresentaram sinais menos graves e se recuperaram espontaneamente. Posteriormente, a planta foi administrada em diferentes épocas a 23 caprinos na dose de 10g/kg. A planta foi significativamente mais tóxica (P<0,05) na época seca; no total, dos 14 caprinos que receberam a planta na época seca, 11 apresentaram sinais clínicos de intoxicação e dos 13 caprinos que receberam a planta na época das chuvas, apenas 3 apresentaram sinais clínicos. Todos os animais que apresentaram sinais clínicos, se recuperaram após a administração de tiosulfato de sódio. Os sinais clínicos caracterizavam-se por apatia, pulso venoso positi-vo, ataxia, berros, taquicardia e taquipneia, midríase e decúbito esternal seguido por decúbito lateral. Antes de cada administração era feito o teste do papel picrosódico para estimar o teor de cianeto na planta, classificando a reação em discreta, leve, moderada e acentuada. As amostras com reação discreta não apresentaram toxicidade, as com reação leve induziram sinais leves e as com reação moderada causaram sinais graves ou moderados de intoxicação. Não foram observados testes com reação acentuada. Os resultados do trabalho demonstram que P. foetida é uma planta cianogênica que causa intoxicação após a ingestão das folhas frescas, principalmente no período de estiagem.

Biologia floral de cinco espécies de Passiflora L. (Passifloraceae) em mata semidecídua

(Biologia floral de cinco espécies de Passiflora L. (Passifloraceae) em mata semidecídua). O estudo da biologia floral de cinco espécies de Passiflora foi feito em uma mata de planalto em Campinas, São Paulo. Passiflora alata, P. amethystina e P. miersii apresentam flores de cor púrpura a violeta e corona variegada. As flores são diurnas, perfumadas, autoincompatíveis e polinizadas por abelhas de grande porte. Passiflora amethystina e P. miersii diferem de P. alata por apresentarem filamentos livres no opérculo, que em P. alata é horizontal e denticulado. Estas diferenças no opérculo promovem comportamentos característicos das abelhas durante as visitas. Passiflora suberosa possui flores verde-amareladas e opérculo plicado. As flores são diurnas, inodoras, autocompatíveis e polinizadas por vespas. Em P. capsularis as flores são brancas e o opérculo é plicado. As flores são noturnas, perfumadas, autocompatíveis e possivelmente polinizadas por mariposas. O opérculo plicado das duas últimas espécies permite que os visitantes tenham fácil acesso ao néctar.

Palinotaxonomia das espécies de Passiflora L. subg. Decaloba (DC.) Rchb. (Passifloraceae) no Sudeste do Brasil

O estudo palinotaxonômico de oito espécies de Passiflora L. subg. Decaloba (DC.) Rchb. (Passifloraceae) teve como objetivo contribuir para a caracterização, circunscrição e delimitação desse subgênero na região Sudeste do Brasil. Os grãos de pólen foram acetolisados, medidos, descritos e ilustrados sob microscopia de luz. Para observar detalhes da superfície e da abertura, grãos de pólen não acetolisados foram analisados em microscópio eletrônico de varredura (MEV) e, em seguida, eletromicrografados. Os dados obtidos foram estatisticamente analisados, de acordo com o tamanho da amostra. As espécies analisadas neste estudo possuem grãos de pólen grandes ou médios, isopolares, prolato-esferoidais, esferoidais ou subprolatos, 12-colporados, 12-colpados para P. suberosa L. ou 6-colporados, com pseudopérculos, exina heteroreticulada. Foi confeccionada uma chave para a identificação das espécies estudadas com base em dados polínicos.

Efeito da taxa de secreção de néctar sobre a polinização e a produção de sementes em flores de Passiflora speciosa Gardn. (Passifloraceae) no Pantanal

Foi determinado o efeito da taxa de secreção de néctar (TSN) sobre a freqüência de visitas dos polinizadores e o número de sementes por flor de Passiflora speciosa Gardn. no Pantanal. A planta foi polinizada por beija-flores e apresentou auto-incompatibilidade. A TSN variou em função do diâmetro da flor, e o número de visitas dos beija-flores foi função do diâmetro da flor. O número de sementes por flor de P. speciosa foi maior com o aumento do número de visitas dos polinizadores. A freqüência de pilhadores de néctar não apresentou efeito sobre a produção de sementes, mas o número total de visitas dos beija-flores foi negativamente correlacionado com o número de visitas dos pilhadores de néctar e/ou pólen. Os resultados indicam que o tamanho da flor afeta a TSN, que por sua vez pode determinar o número de visitas dos polinizadores e o sucesso da produção de sementes em flores de P. speciosa.

Meiotic behavior in wild and domesticated species of Passiflora

The meiotic behavior of fourteen Passiflora taxa was analyzed. The species were grouped according to the n value (6, 9 and 12) for statistical studies. Some species presented tetravalent associations or univalent chromosomes in diakinesis, bivalent formation prevailing. The qui-square test revealed significant differences in the chiasma frequency among species for n = 9 and n = 6 groups. There was predominance of interstitial chiasmata in almost all studied species. The n = 12 group was the only one whose meiotic behavior was considered similar due to the quantity of chiasmata per cell, tendency of interstitial chiasma localization. Some species presented meiotic irregularities, such as laggard and precocious chromosomes in meiosis I. In telophase II the percentages of meiotic irregularities was low. Irregularities in the spindle orientation were presented in higher percentages in the end of meiosis II, and were also responsible for post-meiotic abnormal products. The irregularities observed during meiosis can have influence on the percentage of sterile pollen grains and success of interspecific crossings in Passiflora species.

Self-, cross- and interspecific pollinations in Passiflora capsularis and P. rubra

This study aimed to characterize the reproductive system of Passiflora capsularis L. and P. rubra L. In vivo controlled pollinations, in vitro pollen germination and pollen-ovule (P:O) ratio evaluation were conducted. In self-pollination, intraspecific and interspecific pollination, P. capsularis showed means of 62.5, 68.7 and 48.4% of fertilized flowers, while in P. rubra, the averages were 67.2, 62.5 and 46.9%, respectively. For in vitro germination, 52.2% of P. capsularis pollen grains germinated while in P. rubra, the percentage was 64.4. The P:O ratio was 22.4 for P. capsularis, and 27.4 for P. rubra, which included them in the category of obligatory autogamous. Passiflora capsularis and P. rubra can reproduce both by self-pollination and cross-pollination, and crossings between the two species succeeded though the success rate was lower than 50%. The characteristics of the reproductive system of both species allow the use of greater range of options on breeding methods for production of ornamental Passiflora plants.

Role of mast cell- and non-mast cell-derived inflammatory mediators in immunologic induction of synaptic plasticity

We have previously discovered a long-lasting enhancement of synaptic transmission in mammal autonomic ganglia caused by immunological activation of ganglionic mast cells. Subsequent to mast cell activation, lipid and peptide mediators are released which may modulate synaptic function. In this study we determined whether some mast cell-derived mediators, prostaglandin D2 (PGD2; 1.0 µM), platelet aggregating factor (PAF; 0.3 µM) and U44619 (a thromboxane analogue; 1.0 µM), and also endothelin-1 (ET-1; 0.5 µM) induce synaptic potentiation in the guinea pig superior cervical ganglion (SCG), and compared their effects on synaptic transmission with those induced by a sensitizing antigen, ovalbumin (OVA; 10 µg/ml). The experiments were carried out on SCGs isolated from adult male guinea pigs (200-250 g) actively sensitized to OVA, maintained in oxygenated Locke solution at 37oC. Synaptic potentiation was measured through alterations of the integral of the post-ganglionic compound action potential (CAP). All agents tested caused long-term (LTP; duration ³30 min) or short-term (STP; <30 min) potentiation of synaptic efficacy, as measured by the increase in the integral of the post-ganglionic CAP. The magnitude of mediator-induced potentiation was never the same as the antigen-induced long-term potentiation (A-LTP). The agent that best mimicked the antigen was PGD2, which induced a 75% increase in CAP integral for LTP (antigen: 94%) and a 34% increase for STP (antigen: 91%). PAF-, U44619-, and ET-1-induced increases in CAP integral ranged for LTP from 34 to 47%, and for STP from 0 to 26%. These results suggest that the agents investigated may participate in the induction of A-LTP

Role of immunoglobulin E and mast cells in murine models of asthma

Immunoglobulin E (IgE) and mast cells are believed to play important roles in allergic inflammation. However, their contributions to the pathogenesis of human asthma have not been clearly established. Significant progress has been made recently in our understanding of airway inflammation and airway hyperresponsiveness through studies of murine models of asthma and genetically engineered mice. Some of the studies have provided significant insights into the role of IgE and mast cells in the allergic airway response. In these models mice are immunized systemically with soluble protein antigens and then receive an antigen challenge through the airways. Bronchoalveolar lavage fluid from mice with allergic airway inflammation contains significant amounts of IgE. The IgE can capture the antigen presented to the airways and the immune complexes so formed can augment allergic airway response in a high-affinity IgE receptor (FcepsilonRI)-dependent manner. Previously, there were conflicting reports regarding the role of mast cells in murine models of asthma, based on studies of mast cell-deficient mice. More recent studies have suggested that the extent to which mast cells contribute to murine models of asthma depends on the experimental conditions employed to generate the airway response. This conclusion was further supported by studies using FcepsilonRI-deficient mice. Therefore, IgE-dependent activation of mast cells plays an important role in the development of allergic airway inflammation and airway hyperresponsiveness in mice under specific conditions. The murine models used should be of value for testing inhibitors of IgE or mast cells for the development of therapeutic agents for human asthma.

Differential expression of integrin subunits on adherent and nonadherent mast cells

Mast cell progenitors arise in bone marrow and then migrate to peripheral tissues where they mature. It is presumed that integrin receptors are involved in their migration and homing. In the present study, the expression of various integrin subunits was investigated in three systems of adherent and nonadherent mast cells. Mesentery mast cells, freshly isolated bone marrow-derived mast cells (BMMC) and RBL-2H3 cells grown attached to tissue culture flasks are all adherent mast cells and peritoneal mast cells, and cultured BMMC and RBL-2H3 cells grown in suspension represent nonadherent mast cell populations. Pure populations of mast cells were immunomagnetically isolated from bone marrow, mesentery and peritoneal lavage using the mast cell-specific monoclonal antibody AA4. By immunomicroscopy, we could demonstrate that all of these mast cells expressed alpha4, alpha5, alpha6, ß1 and ß7 integrin subunits. The expression of the alpha4 integrin subunit was 25% higher in freshly isolated mesentery mast cells and BMMC. Consistent with the results obtained by immunomicroscopy, mesentery mast cells expressed 65% more mRNA for the alpha4 integrin subunit than peritoneal mast cells. In vitro studies were also conducted using the rat mast cell line RBL-2H3. RBL-2H3 cells grown attached to the tissue culture flasks or as suspension cultures expressed the same integrin subunits identified in bone marrow, mesenteric and peritoneal mast cells ex vivo. Similarly, the expression of alpha4 integrin was higher in adherent cells. Therefore, alpha4 integrins may play a critical role in the anchorage of mast cells to the extracellular matrix in bone marrow and in peripheral tissues.