The effects of a plant proteinase inhibitor from Enterolobium contortisiliquum on human tumor cell lines

Supplementary to the efficient inhibition of trypsin, chymotrypsin, plasma kallikrein, and plasmin already described by the EcTI inhibitor from Enterolobium contortisiliquum, it also blocks human neutrophil elastase (K(iapp)=4.3 nM) and prevents phorbol ester (PMA)-stimulated activation of matrix metalloproteinase (MMP)-2 probably via interference with membrane-type 1 (MT1)-MMP. Moreover, plasminogen-induced activation of proMMP-9 and processing of active MMP-2 was also inhibited. Furthermore, the effect of EcTI on the human cancer cell lines HCT116 and HT29 (colorectal), SkBr-3 and MCF-7 (breast), K562 and THP-1 (leukemia), as well as on human primary fibroblasts and human mesenchymal stem cells (hMSCs) was studied. EcTI inhibited in a concentration range of 1.0-2.5 mu M rather specifically tumor cell viability without targeting primary fibroblasts and hMSCs. Taken together, our data indicate that the polyspecific proteinase inhibitor EcTI prevents proMMP activation and is cytotoxic against tumor cells without affecting normal tissue remodeling fibroblasts or regenerative hMSCs being an important tool in the studies of tumor cell development and dissemination

Use of visual and olfactory cues of flowers of two brassicaceous species by insect pollinators

1. Pollinating insects exploit visual and olfactory cues associated with flower traits indicative of flower location and reward quality. Pollination can induce changes in these flower-associated cues, thereby influencing the behaviour of flower visitors. 2. This study investigated the main cues exploited by the syrphid fly Episyrphus balteatus and the butterfly Pieris brassicae when visiting flowers of Brassica nigra and Raphanus sativus plants. Whether pollen is used as a cue and whether pollination-induced changes affect flower volatile emission and the behavioural responses of the two pollinator species were also studied. 3. Pollinator preference was investigated by offering visual and olfactory cues individually as well as simultaneously in two-choice bioassays. Plant treatments included emasculation, hand-pollination and untreated control plants. The composition of flower volatiles from pollinated and unpollinated control plants was analysed by gas chromatography-mass spectrometry. 4. Both pollinators exhibited a strong bias for visual cues over olfactory cues. Neither pollinator used pollen as a cue. However, E. balteatus discriminated between newly opened and long-open flowers at short distance only when pollen was available. Flower visits by pollinators were influenced by pollination-induced changes in B. nigra but not R. sativus flowers. Pieris brassicae only responded to pollination-induced changes when visual and olfactory cues were offered simultaneously. The blend of volatiles emitted by B. nigra, but not R. sativus inflorescences was affected by pollination. 5. Collectively, the findings of this study show that different pollinators exploit different visual and olfactory traits when searching for flowers of two brassicaceous plant species.</p

Endure and call for help : Strategies of black mustard plants to deal with a specialized caterpillar

Plants have evolved inducible resistance and tolerance mechanisms against insect herbivores. Resistance mechanisms that affect herbivorous insects directly can be effective against generalist herbivores, but will not deter specialist herbivores from attacking the plant. Tolerance mechanisms and indirect plant resistance are more likely effective strategies used by plants when dealing with specialist herbivores. However, inducible indirect resistance and tolerance mechanisms have rarely been investigated within the same study system. We studied multiple tolerance mechanisms and the role of natural enemies in reducing fitness loss of Brassica nigra plants incurred by feeding by the specialist herbivore Pieris brassicae. For this, we measured the changes in carbon and nitrogen triggered by herbivore attack and quantified plant biomass and seed production under field conditions, in the presence or absence of natural enemies of the herbivores. We also assessed whether B. nigra increased selfing rates when exposed to herbivore infestation, and investigated whether infestation by P. brassicae rendered B. nigra plants more attractive to night-active pollinators than control plants. We found that B. nigra flowers are rarely visited by insects during the night, and exposure to herbivores did not influence selfing rates. Brassica nigra plants compensated for herbivory in terms of vegetative biomass. Seed set was negatively affected by herbivory in the absence of natural enemies, but not in the presence of natural enemies. Plants responded to herbivory with drastic changes in nitrogen contents of leaves and flowers, whereas no changes in carbon concentrations were detected. The investment in reproduction or re-growth of vegetative tissues is not sufficient to sustain plant fitness. Reproductive output of flowering mustard plants is only sustained when interactions with the natural enemies of the herbivores are preserved. We conclude that natural enemies of herbivorous insects play an important role as component of the plant's defence strategy. This study reveals that both tolerance and indirect resistance are strategies of this plant species to sustain fitness

Effects of Escherichia coli LPS Structure on Antibacterial and Anti-Endotoxin Activities of Host Defense Peptides

The binding of Host Defense Peptides (HDPs) to the endotoxin of Gram-negative bacteria has important unsolved aspects. For most HDPs, it is unclear if binding is part of the antibacterial mechanism or whether LPS actually provides a protective layer against HDP killing. In addition, HDP binding to LPS can block the subsequent TLR4-mediated activation of the immune system. This dual activity is important, considering that HDPs are thought of as an alternative to conventional antibiotics, which do not provide this dual activity. In this study, we systematically determine, for the first time, the influence of the O-antigen and Lipid A composition on both the antibacterial and anti-endotoxin activity of four HDPs (CATH-2, PR-39, PMAP-23, and PMAP36). The presence of the O-antigen did not affect the antibacterial activity of any of the tested HDPs. Similarly, modification of the lipid A phosphate (MCR-1 phenotype) also did not affect the activity of the HDPs. Furthermore, assessment of inner and outer membrane damage revealed that CATH-2 and PMAP-36 are profoundly membrane-active and disrupt the inner and outer membrane of Escherichia coli simultaneously, suggesting that crossing the outer membrane is the rate-limiting step in the bactericidal activity of these HDPs but is independent of the presence of an O-antigen. In contrast to killing, larger differences were observed for the anti-endotoxin properties of HDPs. CATH-2 and PMAP-36 were much stronger at suppressing LPS-induced activation of macrophages compared to PR-39 and PMAP-23. In addition, the presence of only one phosphate group in the lipid A moiety reduced the immunomodulating activity of these HDPs. Overall, the data strongly suggest that LPS composition has little effect on bacterial killing but that Lipid A modification can affect the immunomodulatory role of HDPs. This dual activity should be considered when HDPs are considered for application purposes in the treatment of infectious diseases.</p

Role of hydrocarbons in egg recognition in the honeybee

Despite worker-laid and queen-laid honeybee eggs having significantly different hydrocarbon profiles, bioassays and chemical supplementation studies show that changing the hydrocarbon profile does not affect egg identity. Furthermore, full-sized eggs that are tested just before being laid or just after being laid have similar hydrocarbon patterns but are treated differently in egg-discrimination bioassays with only the laid eggs surviving. This suggests that hydrocarbons play no role in the egg-recognition system in honeybees