Enterolobium contortisiliquum trypsin inhibitor (EcTI), a plant proteinase inhibitor, decreases in vitro cell adhesion and invasion by inhibition of Src protein-focal adhesion kinase (FAK) signaling pathways

Tumor cell invasion is vital for cancer progression and metastasis. Adhesion, migration, and degradation of the extracellular matrix are important events involved in the establishment of cancer cells at a new site, and therefore molecular targets are sought to inhibit such processes. The effect of a plant proteinase inhibitor, Enterolobium contortisiliquum trypsin inhibitor (EcTI), on the adhesion, migration, and invasion of gastric cancer cells was the focus of this study. EcTI showed no effect on the proliferation of gastric cancer cells or fibroblasts but inhibited the adhesion, migration, and cell invasion of gastric cancer cells; however, EcTI had no effect upon the adhesion of fibroblasts. EcTI was shown to decrease the expression and disrupt the cellular organization of molecules involved in the formation and maturation of invadopodia, such as integrin β1, cortactin, neuronal Wiskott-Aldrich syndrome protein, membrane type 1 metalloprotease, and metalloproteinase-2. Moreover, gastric cancer cells treated with EcTI presented a significant decrease in intracellular phosphorylated Src and focal adhesion kinase, integrin-dependent cell signaling components. Together, these results indicate that EcTI inhibits the invasion of gastric cancer cells through alterations in integrin-dependent cell signaling pathways.     

Baupain, a plant cysteine proteinase that hinders thrombin-induced human platelet aggregation

Bauninia forficata is trivially known as cow paw, and popularly used in Brazil for treatment of diabetes mellitus. Denominated baupain a cysteine proteinase was purified from B. forficata leaves. In this study, we investigated the baupain effect on aggregation of isolated human platelets in vitro and the results show that baupain hinders thrombin - but not ADP- and collagen- induced platelet aggregation. With synthetic quenched-fluorescent peptides, the kinetics of the cleavage site of human proteinase-activated receptor 1 / 2 / 3 and 4 [PAR-1 / 2 / 3 and 4] by baupain was determined. In conclusion, similar to bromelain and papain, baupain hinders human platelets aggregation, probably through an unspecific cleavage in the Phe-Leu bond of PAR1.     

Effects of compounds from Passiflora edulis Sims f. flavicarpa juice on blood coagulation and on proteolytic enzymes

Passion fruit (Passiflora edulis Sims f. flavicarpa) is popularly known for its sedative and calming properties and is consumed as a fresh fruit or as a juice. The clinical observation of blood incoagulability associated with excessive consumption of passion fruit juice, in a patient treated with warfarin, prompted the current study to investigate in vitro the presence of blood clotting inhibitors in Passiflora edulis Sims f. flavicarpa extract. After purification process, two compounds of distinct molecular weight and inhibitory action were better characterized. One is a trypsin inhibitor similar to inhibitors from Bowman-Birk family, named PeTI-I12, and other is a compound active in coagulation that prolongs aPTT and PT, but does not change TT. The aim of this study is to provide evidence that passion fruit extract's components play a role on hemostasis and therefore may be relevant in the handling of patients treated with anticoagulants or suffering hemorrhagic diseases.     

A new species of Oxelytrum Gistel (Coleoptera, Silphidae) from southern Argentina, with a key to the species of the genus

A new species of the forensically interesting genus Oxelytrum Gistel (Coleoptera, Silphidae), Oxelytrum selknan, is described from Santa Cruz and Tierra del Fuego provinces, Argentina. The new species resembles Oxelytrum biguttatum (Philippi) in outer aspect, but has different male genitalia, in particular a median lobe longer than the paramera. All the described species of Oxelytrum have the median lobe shorter than the paramera. The internal sac, as far as it could be reconstructed from dry-pinned specimens, also shows differences between the two species. A key to the species of Oxelytrum is given and illustrated.     

A plant Kunitz-type inhibitor mimics bradykinin-induced cytosolic calcium increase and intestinal smooth muscle contraction

Abstract: BbKI is a kallikrein inhibitor with a reactive site sequence similar to that of kinins, the vasoactive peptides inserted in kininogen moieties. This structural similarity probably contributes to the strong interaction with plasma kallikrein, the enzyme that releases, from high-molecular weight kininogen (HMWK), the proinflammatory peptide bradykinin, which acts on B2 receptors (B2R). BbKI was examined on smooth muscle contraction and Ca2+ mobilization, in which the kallikrein-kinin system is involved. Contrary to expectations, BbKI (1.8 μm) increased [Ca2+]cand contraction, as observed with BK (2.0 μm). Not blocked by B1 receptors (B1R), the BbKI agonistic effect was blocked by the B2R antagonist, HOE-140 (6 μm), and the involvement of B2R was confirmed in B2R-knockout mice intestine. The same tissue response was obtained using a synthetic peptide derived from the BbKI reactive site structure, more resistant than BK to angiotensin I-converting enzyme (ACE) hydrolysis. Depending on the concentration, BbKI has a dual effect. At a low concentration, BbKI acts as a potent kallikrein inhibitor; however, due to the similarity to BK, in high concentrations, BbKI greatly increases Ca2+ release from internal storages, as a consequence of its interaction with B2R. Therefore, the antagonistic and agonistic effects of BbKI may be considered in conditions of B2R involvement.     

TNF-α is associated with loss of lean body mass only in already cachectic COPD patients

Background

Systemic inflammation may contribute to cachexia in patients with chronic obstructive pulmonary disease (COPD). In this longitudinal study we assessed the association between circulating C-reactive protein (CRP), tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6 levels and subsequent loss of fat free mass and fat mass in more than 400 COPD patients over three years.

Methods

The patients, aged 40–76, GOLD stage II-IV, were enrolled in 2006/07, and followed annually. Fat free mass and fat mass indexes (FFMI & FMI) were calculated using bioelectrical impedance, and CRP, TNF-α, IL-1ß, and IL-6 were measured using enzyme immunoassays. Associations with mean change in FFMI and FMI of the four inflammatory plasma markers, sex, age, smoking, FEV₁, inhaled steroids, arterial hypoxemia, and Charlson comorbidity score were analyzed with linear mixed models.

Results

At baseline, only CRP was significantly (but weakly) associated with FFMI (r = 0.18, p < 0.01) and FMI (r = 0.27, p < 0.01). Univariately, higher age, lower FEV₁, and use of beta2-agonists were the only significant predictors of decline in FFMI, whereas smoking, hypoxemia, Charlson score, and use of inhaled steroids predicted increased loss in FMI. Multivariately, high levels of TNF-α (but not CRP, IL-1ß or IL-6) significantly predicted loss of FFMI, however only in patients with established cachexia at entry.

Conclusion

This study does not support the hypothesis that systemic inflammation is the cause of accelerated loss of fat free mass in COPD patients, but suggests a role for TNF-α in already cachectic COPD patients.     

Extending signaling pathways with protein-interaction networks. Application to apoptosis

Cells exploit signaling pathways during responses to environmental changes, and these processes are often modulated during disease. Particularly, relevant human pathologies such as cancer or viral infections require downregulating apoptosis signaling pathways to progress. As a result, the identification of proteins responsible for these changes is essential for the diagnostics and development of therapeutics. Transferring functional annotation within protein interaction networks has proven useful to identify such proteins, although this is not a trivial task. Here, we used different scoring methods to transfer annotation from 53 well-studied members of the human apoptosis pathways (as known by 2005) to their protein interactors. All scoring methods produced significant predictions (compared to a random negative model), but its number was too large to be useful. Thus, we made a final prediction using specific combinations of scoring methods and compared it to the proteins related to apoptosis signaling pathways during the last 5 years. We propose 273 candidate proteins that may be relevant in apoptosis signaling pathways. Although some of them have known functions consistent with their proposed apoptotsis involvement, the majority have not been annotated yet, leaving room for further experimental studies. We provide our predictions at http://sbi.imim.es/web/Apoptosis.php.     

What can metazoan parasites reveal about the taxonomy of Scomber japonicus Houttuyn in the coast of South America and Madeira Islands?

The metazoan parasites of four populations of the chub mackerel Scomber japonicus were analysed from two localities in the Atlantic Ocean (Madeira Islands, Portugal, and Rio de Janeiro, Brazil) and two localities in the Pacific Ocean (Callao, Peru, and Antofagasta, Chile), collected during 2002 and 2003. A total of 373 fish specimens were studied and 34 metazoan parasite species were obtained. Parasites identified from the populations of chub mackerel studied could be separated into three categories: parasites with a wide distribution, present in the Pacific and Atlantic, parasites proper of the Pacific Ocean and parasites proper of the Atlantic Ocean. The analyses of some highly specific parasites of the genus Scomber ( i.e. monogeneans of the genus Kuhnia and didymozoid digeneans) strongly suggest the need for a revision of the taxonomic status of chub mackerels from the Atlantic and Pacific coast of America. The results demonstrated the usefulness of parasites as adequate tools to clarify the taxonomic status of their hosts.     

Photosynthetic limitations in leaves of young Brazilian Green Dwarf coconut (Cocos nucifera L. ‘nana’) palm under well-watered conditions or recovering from drought stress

Young plants of two Brazilian Green Dwarf coconut ecotypes from contrasting regions in relation to climate (UGD, from a hot and humid climate and JGD, from a hot and dry climate) were submitted to three consecutive drying/recovery cycles, under greenhouse conditions to determine the photosynthetic limitations encountered during the recovery phase of water deficiency. Three hypotheses were tested: (1) non-stomatal factors contribute to the incomplete recovery of net photosynthesis rate (A), (2) photochemical impairment is an important component of the non-stomatal limitations to A and (3) the two coconut ecotypes respond differently to the drought recovery. In each drying/recovery cycle, irrigation was suppressed until A reached zero. Then the plants were rewatered and the recovery was accompanied for 8 days. After 4 days of rewatering, CO₂ and light response curve parameters were used to discriminate the stomatal and non-stomatal factors contributing to the incomplete recovery of A. Upon rewatering for 4 days, the pre-dawn leaf water potential of the stressed plants increased from ?1.20 to ?0.15 MPa on average. Only small differences were detected in the intercellular to atmospheric CO₂ concentration ratio but A did not recover completely to the control values. In both ecotypes, light and CO₂-saturated A, carboxylation efficiency, maximum quantum yield of PSII and maximum electron transport rate were significantly lower in stressed plants in comparison with the well-watered ones. Relative stomatal limitation (L_S) ranged from 16 to 20% without significant differences between treatments and between ecotypes. The relative mesophyll limitation (L_m) was significantly higher in JGD in cycle 1 but this changed after repeated drought stress, decreasing to values significantly lower than those in UGD in the other two cycles. The results showed that after 4 days of rewatering stressed plant stomatal limitations were comparable to that of well-watered control plants (～20%). However A did not recover completely after drought stress due to non-stomatal factors. Important differences between the ecotypes were identified. The ecotype JGD presented smaller L_m, faster recovery of the apparent quantum efficiency of CO₂ assimilation, adjustment of some carbon balance components and signs of photosynthetic capacity acclimation after repeated drying/recovery cycles.