Biochemical and enzymatic characterization of BpMP-I, a fibrinogenolytic metalloproteinase isolated from Bothropoides pauloensis snake venom

Snake Venom Metalloproteinases (SVMPs) are the most abundant components present in Viperidae venom. They are important in the induction of systemic alterations and local tissue damage after envenomation. In the present study, a metalloproteinase named BpMPI was isolated from Bothropoides pauloensis snake venom and its biochemical and enzymatic characteristics were determined. BpMPI was purified in two chromatography steps on ion exchange CM-Sepharose Fast flow and Sephacryl S-300. This protease was homogeneous on SDS-PAGE and showed a single chain polypeptide of 20 kDa under non reducing conditions. The partial amino acid sequence of the enzyme showed high similarity with other SVMPs enzymes from snake venoms. BpMPI showed proteolytic activity upon azocasein and bovine fibrinogen and was inhibited by EDTA, 1,10 phenanthroline and β-mercaptoethanol. Moreover, this enzyme showed stability at neutral and alkaline pH and it was inactivated at high temperatures. BpMPI was able to hydrolyze glandular and tissue kallikrein substrates, but was unable to act upon factor Xa and plasmin substrates. The enzyme did not induce local hemorrhage in the dorsal region of mice even at high doses. Taken together, our data showed that BpMP-I is in fact a fibrinogenolytic metalloproteinase and a non hemorrhagic enzyme.     

Street heroin induces mitochondrial dysfunction and apoptosis in rat cortical neurons

Cortical function has been suggested to be highly compromised by repeated heroin self-administration. We have previously shown that street heroin induces apoptosis in neuronal-like PC12 cells. Thus, we analysed the apoptotic pathways involved in street heroin neurotoxicity using primary cultures of rat cortical neurons. Our street heroin sample was shown to be mainly composed by heroin, 6-monoacetylmorphine and morphine. Exposure of cortical neurons to street heroin induced a slight decrease in metabolic viability, without loss of neuronal integrity. Early activation of caspases involved in the mitochondrial apoptotic pathway was observed, culminating in caspase 3 activation, Poly-ADP Ribose Polymerase (PARP) cleavage and DNA fragmentation. Apoptotic morphology was completely prevented by the non-selective caspase inhibitor z-VAD-fmk, indicating an important role for caspases in neurodegeneration induced by street heroin. Ionotropic glutamate receptors, opioid receptors and oxidative stress were not involved in caspase 3 activation. Interestingly, street heroin cytotoxicity was shown to be independent of a functional mitochondrial respiratory chain, as determined using NT-2 rho(0) cells. Nonetheless, in street heroin-treated cortical neurons, cytochrome c was released, accompanied by a decrease in mitochondrial potential and Bcl-2/Bax. Pure heroin hydrochloride similarly decreased metabolic viability but only slightly activated caspase 3. Altogether, our data suggest an important role for mitochondria in mediating street heroin neurotoxic effects.     

Development of a novel anti-biofilm peptide derived from profilin of Spodoptera frugiperda

Abstract

Candida yeast infections are the fourth leading cause of death worldwide. Peptides with antimicrobial activity are a promising alternative treatment for such infections. Here, the antifungal activity of a new antimicrobial peptide—PEP-IA18—was evaluated against Candida species. PEP-IA18 was designed from the primary sequence of profilin, a protein from Spodoptera frugiperda, and displayed potent activity against Candida albicans and Candida tropicalis, showing a minimum inhibitory concentration (MIC) of 2.5?µM. Furthermore, the mechanism of action of PEP-IA18 involved interaction with the cell membrane (ergosterol complexation). Treatment at MIC and/or 10?×?MIC significantly reduced biofilm formation and viability. PEP-IA18 showed low toxicity toward human fibroblasts and only revealed hemolytic activity at high concentrations. Thus, PEP-IA18 exhibited antifungal and anti-biofilm properties with potential applicability in the treatment of infections caused by Candida species.     

RETRACTED ARTICLE: Mast cells are the main interleukin 17-positive cells in anticitrullinated protein antibody-positive and -negative rheumatoid arthritis and osteoarthritis synovium

Introduction  

Mast cells have been implicated to play a functional role in arthritis, especially in autoantibody-positive disease. Among the cytokines involved in rheumatoid arthritis (RA), IL-17 is an important inflammatory mediator. Recent data suggest that the synovial mast cell is a main producer of IL-17, although T cells have also been implicated as prominent IL-17 producers as well. We aimed to identify IL-17 expression by mast cells and T cells in synovium of arthritis patients.     

Reproductive isolation between diploid and tetraploid cytotypes of Libidibia ferrea (= Caesalpinia ferrea) (Leguminosae): ecological and taxonomic implications

Polyploidy is the most common chromosomal mechanism involved in the evolution of plants. However, the emergence of polyploid individuals does not guarantee the establishment of a new lineage, and the relationship between polyploidy and reproductive biology is therefore relevant. Libidibia ferrea is a legume tree that has diploid and tetraploid populations. In this work we analyse the reproductive biology of the species to verify the degree of reproductive isolation between the two cytotypes. Observations on phenology, floral morphology, biology, and visitors, breeding system, reproductive success and reproductive isolation were made for both cytotypes in two municipalities of northeastern Brazil. Cytotypes differed for all morphometric parameters analysed, with tetraploids exhibiting higher mean values than diploids. Both cytotypes had the same effective pollinators (native bees from the genera Centris and Xylocopa, and the introduced Apis mellifera). However, since stamens of diploids and tetraploids were of different size, it is possible that spatial separation occurs when pollen of each cytotype is deposited on a bee’s body. Diploids were self-incompatible (ISI?=?0) and exhibited high fruit-set after intraploidy crossing (20?%), whereas tetraploids were self-compatible (ISI?=?0.47) and set only 3.9?% of fruits after intraploidy crossing. Both cytotypes showed low fruit- and seed-set under natural conditions, and there was no fruit-set after crosses between them. Data are discussed in relation to the establishment of polyploids, to the pattern of distribution of the species and to the taxonomic implications. The cytotypes of L. ferrea should be considered as distinct species, since they are reproductively isolated from each other.     

Review of the large catfish fisheries in the upper Amazon and the stock depletion of <Emphasis Type="Italic">piraíba</Emphasis> (<Emphasis Type="Italic">Brachyplatystoma filamentosum</Emphasis>Lichtenstein)

In this paper we describe the present status of the large migratory catfish fisheries in the Upper Amazon. We present biological information about the main species and we give strong evidence that the stock of piraíba (Brachyplatystoma filamentosum),the largest catfish in the Amazon Basin is probably over-exploited. In conclusion, we raise some hypotheses about the causes and prospects for the future.     

Route of jejunal mucosa absorption of trypsin demonstrated by immunofluorescence

Previous studies have demonstrated the absorption of porcine trypsin in isolated jejunal loops from male Wistar rats by open-loop perfusion. The possible routes of absorption were examined in the study reported here. Trypsin (0.5 mg/ml) was dissolved in tyrode solution and perfused at a rate of 0.5 ml/min, at 37 degrees C, for 40 min. Using immunoperoxidase and immunofluorescence techniques, strong reactivity towards anti-TLCK-trypsin antibody was demonstrated through out the enterocyte cytosol. The present data indicate that trypsin was absorbed by enterocytes, probably through a transcellular route.     

Clinical presentations of nosocomial aspergillosis

Aspergillus is a very ubiquitous fungal in our environment, including hospitals, being the second in frequency in colonization and infection, just after Candida spp. Aspergillus nosocomial infections have increased, because the number of immunocompromised patients has also increased. Nosocomial infections can be caused by different species of Aspergillus, being pulmonary manifestations the most frequent. Primary or secondary nonpulmonary infections can affect the brain, heart, kidney, eyes and other organs. The mortality due to invasive aspergillosis is very high, and a clinical-radiological suspicion and, specially the instauration of a rapid treatment with high doses of amphotericin B or its new formulations (associated with surgery in many times) may modify the mortality observed in this patients.