State of the Art in the Studies on Crotamine, a Cell Penetrating Peptide from South American Rattlesnake

Animal venoms comprise a naturally selected cocktail of bioactive peptides/proteins and other molecules, each of which playing a defined role thanks to the highly specific interactions with diverse molecular targets found in the prey. Research focused on isolation, structural, and functional characterizations of novel natural biologics (bioactive peptides/proteins from natural sources) has a long way to go through from the basic science to clinical applications. Herein, we overview the structural and functional characteristics of the myoneurotoxin crotamine, firstly isolated from the South American rattlesnake venom. Crotamine is the first venom peptide classified as a natural cell penetrating and antimicrobial peptide (CPP and AMP) with a more pronounced antifungal activity. in contrast to other known natural CPPs and AMPs, crotamine demonstrates a wide spectrum of biological activities with potential biotechnological and therapeutic values. More recent studies have demonstrated the selective in vitro anticancer activity of crotamine. in vivo, using a murine melanoma model, it was shown that crotamine delays tumor implantation, inhibits tumor cells proliferation, and also increases the survival of mice engrafted with subcutaneous melanoma. the structural and functional properties and also the possible biotechnological applications of minimized molecules derived from crotamine are also discussed.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Inst Butantan, Genet Lab, BR-05503900 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Farmacol, São Paulo, BrazilUniv Fed Ceara, Labomar Inst Ciencias Mar, Fortaleza, CE, BrazilUniv Estado Amazonas, Manaus, AM, BrazilCBA, Lab Bioquim & Biol Mol, Manaus, AM, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Farmacol, São Paulo, BrazilWeb of Scienc

Preliminary in vitro evaluation of N '-(benzofuroxan-5-yl)methylene benzohydrazide derivatives as potential anti-Trypanosoma cruzi agents

A set of benzofuroxan derivatives was tested in vitro against Trypanosoma cruzi epimastigote forms. The influence of physicochemical properties on these benzofuroxan derivatives' activity was observed, and the presence of electron-withdrawing and hydrophobic groups attached to the benzene ring seems to make a favorable contribution at lower concentrations.CNPqCNPqCAPESCAPESFAPESPFAPES

Efeito citotoxico do amblyomin-X em celulas tumorais de melanomase e de adenocarcinoma de pancreas

BV UNIFESP: Teses e dissertaçõe

Mastoparan induces apoptosis in B1 6F10-Nex2 melanoma cells via the intrinsic mitochondrial pathway and displays antitumor activity in vivo

Mastoparan is an a-helical and amphipathic tetradecapeptide obtained from the venom of the wasp Vespula lewisii. This peptide exhibits a wide variety of biological effects, including antimicrobial activity, increased histamine release from mast cells, induction of a potent mitochondrial permeability transition and tumor cell cytotoxicity. Here, the effects of mastoparan in malignant melanoma were studied using the murine model of B16F10-Nex2 cells. in vitro, mastoparan caused melanoma cell death by the mitochondrial apoptosis pathway, as evidenced by the Annexin V-FITC/PI assay, loss of mitochondrial membrane potential (Delta psi(m)), generation of reactive oxygen species, DNA degradation and cell death signaling. Most importantly, mastoparan reduced the growth of subcutaneous melanoma in syngeneic mice and increased their survival. the present results show that mastoparan induced caspase-dependent apoptosis in melanoma cells through the intrinsic mitochondrial pathway protecting the mice against tumor development. (C) 2014 Elsevier Inc. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)INCT ToxConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Butantan Inst, Biochem & Biophys Lab, São Paulo, SP, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Expt Oncol Unit UNONEX, São Paulo, SP, BrazilUniv São Paulo, Inst Biomed Sci, Dept Immunol, BR-05508 São Paulo, SP, BrazilUniv São Paulo, Cell & Mol Therapy Ctr NUCEL NETCEM, São Paulo, BrazilUniv São Paulo, Sch Med, Lab Genet & Mol Hematol LIM31, BR-05508 São Paulo, BrazilButantan Inst, Lab Genet, São Paulo, SP, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Expt Oncol Unit UNONEX, São Paulo, SP, BrazilFAPESP: 2010/51077-5Web of Scienc

BFD-22 a new potential inhibitor of BRAF inhibits the metastasis of B16F10 melanoma cells and simultaneously increased the tumor immunogenicity

Benzofuroxan is an interesting ring system, which has shown a wide spectrum of biological responses against tumor cell lines. We investigated, herein, the antitumor effects of benzofuroxan derivatives (BFDs) in vitro and in a melanoma mouse model. Cytotoxic effects of twenty-two BFDs were determined by MIT assay. Effects of BFD-22 in apoptosis and cell proliferation were evaluated using Annexin V-FITC/PI and CFSE staining. In addition, the effects in the cell cycle were assessed. Flow cytometry, western blot, and fluorescence microscopy analysis were employed to investigate the apoptosis-related proteins and the BRAF signaling. Cell motility was also exploited through cell invasion and migration assays. Molecular docking approach was performed in order to verify the BFD-22 binding mode into the ATP catalytic site of BRAF kinase. Moreover, the BFD-22 antitumor effects were evaluated in a melanoma murine model using B16F10. BFD-22 was identified as a potential hit against melanoma cells. BFD-22 induced apoptosis and inhibited cell proliferation of B16F10 cells. BFD-22 has suppressed, indeed, the migratory and invasive behavior of B16F10 cells. Cyclin D1 and CDK4 expression were reduced leading to cell cycle arrest at G0/G1 phase. Of note, phosphorylation of BRAF at Ser338 was strongly down-regulated by BFD-22 in B16F10 cells. The accommodation/orientation into the binding site of BRAF was similar of BAY43-9006 (co-crystallized inhibitor of BRAF, sorafenib). Importantly, BFD-22 presented in vivo antimetastatic effects and showed better therapeutic efficacy than sorafenib and taxol. BFD-22 can be considered as a new lead compound and, then, can be helpful for the designing of novel drug candidates to treat melanoma. (C) 2016 Elsevier Inc. All rights reserved.Sao Paulo Research Foundation (FAPESP)Brazilian Governmental Agency CNPqBrazilian Governmental Agency CAPESUniv Sao Paulo, Lab Tumor Immunol, Sao Paulo, SP, BrazilButantan Inst, Biochem & Biophys Lab, Sao Paulo, SP, BrazilUniv Groningen, Univ Med Ctr Groningen, Dept Med Oncol, Hanzepl 1, Groningen, NetherlandsUniv Sao Paulo, Lab Drug Design & Dev, Sao Paulo, SP, BrazilUniv Sao Paulo, Lab Cytopathol, Dept Clin Chem & Toxicol, Fac Pharmaceut Sci, Sao Paulo, SP, BrazilButantan Inst, Genet Lab, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Expt Oncol Sect, Sao Paulo, SP, BrazilUniv Sao Paulo, Cell & Mol Therapy Ctr NUCEL NETCEM, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Expt Oncol Sect, Sao Paulo, SP, BrazilFAPESP: 2009/54599-5FAPESP: 2013/07273-2FAPESP: 2013/05396-0FAPESP: 2014/07341-0FAPESP: 2014/14267-1Web of Scienc

General aspects of the rebio-jaru Amazon forest micrometeorological tower LBA wet season campaign and preliminary results

A description of the 60m height micrometeorological tower built at the Amazonian Forest Rebio-Jaru Reserve (10 04' S; 61 56' W) during the LBA (Large Scale Biosphere Atmosphere Experiment in Amazonia) wet season campaign (January to March 1999), in the Brazilian State of Rondonia, is presented. Also, some of the preliminary results obtained during the campaign are presented. The Amazonian forest canopy has a mean height of 35m; however, some of the higher tree branches have heights up to 45m. So, to start the turbulent exchange process studies in this site, it is important to admit that there is a thermodynamic canopy height of about 35m, and another aerodynamic of about 45m. These facts were considered to decide the appropriate height at which the instruments were placed on the tower. Fast response instruments (3D sonic-anemometers, thermometers and hygrometers), with a sampling rate of 16 Hz (except during three days, when the sampling rate was 60Hz), were installed. Also, slow response instruments, operated at a sampling rate of 0.1Hz, provided vertical profiles of the following meteorological variables: wind velocity, temperature, specific humidity, incoming solar radiation, net radiation and photosynthetically active radiation (PAR). One of the goals of this investigation was to analyse the dependence of the vertical eddy fluxes on the cutoff time scale, as well as on the flux averaging time scale. In this sense, graphs of composite averaged fluxes and their changes with increasing cutoff scale, for several hours in a typical convective day in Amazonia are presented. They are similar to the ones presented by Sun et al (1). The results show that the flux averaging scale needed to stabilize the flux calculation changes along the day, and reaches a value greater than 20min around noon time. Some typical patterns of the diurnal variability of the measured micrometeorological variables above and inside the forest canopy are also presented. These results explain some of the physical aspects of the time variability patterns in the Amazonian Forest on a wet day

Organizational factors associated with adherence to low tidal volume ventilation: a secondary analysis of the CHECKLIST-ICU database

Background: Survival benefit from low tidal volume (VT) ventilation (LTVV) has been demonstrated for patients with acute respiratory distress syndrome (ARDS), and patients not having ARDS could also benefit from this strategy. Organizational factors may play a role on adherence to LTVV. The present study aimed to identify organizational factors with an independent association with adherence to LTVV. Methods: Secondary analysis of the database of a multicenter two-phase study (prospective cohort followed by a cluster-randomized trial) performed in 118 Brazilian intensive care units. Patients under mechanical ventilation at day 2 were included. LTVV was defined as a VT ≤ 8 ml/kg PBW on the second day of ventilation. Data on the type and number of beds of the hospital, teaching status, nursing, respiratory therapists and physician staffing, use of structured checklist, and presence of protocols were tested. A multivariable mixed-effect model was used to assess the association between organizational factors and adherence to LTVV. Results: The study included 5719 patients; 3340 (58%) patients received LTVV. A greater number of hospital beds (absolute difference 7.43% [95% confidence interval 0.61–14.24%]; p = 0.038), use of structured checklist during multidisciplinary rounds (5.10% [0.55–9.81%]; p = 0.030), and presence of at least one nurse per 10 patients during all shifts (17.24% [0.85–33.60%]; p = 0.045) were the only three factors that had an independent association with adherence to LTVV. Conclusions: Number of hospital beds, use of a structured checklist during multidisciplinary rounds, and nurse staffing are organizational factors associated with adherence to LTVV. These findings shed light on organizational factors that may improve ventilation in critically ill patients

Identification of human chromosome 22 transcribed sequences with ORF expressed sequence tags

Transcribed sequences in the human genome can be identified with confidence only by alignment with sequences derived from cDNAs synthesized from naturally occurring mRNAs. We constructed a set of 250,000 cDNAs that represent partial expressed gene sequences and that are biased toward the central coding regions of the resulting transcripts. They are termed ORF expressed sequence tags (ORESTES). The 250,000 ORESTES were assembled into 81,429 contigs. Of these, 1,181 (1.45%) were found to match sequences in chromosome 22 with at least one ORESTES contig for 162 (65.6%) of the 247 known genes, for 67 (44.6%) of the 150 related genes, and for 45 of the 148 (30.4%) EST-predicted genes on this chromosome. Using a set of stringent criteria to validate our sequences, we identified a further 219 previously unannotated transcribed sequences on chromosome 22. Of these, 171 were in fact also defined by EST or full length cDNA sequences available in GenBank but not utilized in the initial annotation of the first human chromosome sequence. Thus despite representing less than 15% of all expressed human sequences in the public databases at the time of the present analysis, ORESTES sequences defined 48 transcribed sequences on chromosome 22 not defined by other sequences. All of the transcribed sequences defined by ORESTES coincided with DNA regions predicted as encoding exons by genscan. (http://genes.mit.edu/GENSCAN.html)