Exoantigen test for identification of Petriellidium boydii cultures

Cultures of Petriellidium boydii were serologically identified by detection of their exoantigens with an immunodiffusion procedure. The technique, which is specific and sensitive, allowed the rapid identification and differentiation of 12 isolates of P. boydii from numerous other morphologically similar Hyphomycetes. The antigen-antiserum reference system and the production, by two different techniques, of exoantigens used in the identification of P. boydii are described

VisiStat: visualization-driven, interactive statistical analysis

Short synthetic peptides corresponding to sequences of complementarity-determining regions (CDRs) from different immunoglobulin families have been shown to induce antimicrobial, antiviral and antitumor activities regardless of the specificity of the original monoclonal antibody (mAb). Presently, we studied the in vitro and in vivo antitumor activity of synthetic peptides derived from conserved CDR sequences of different immunoglobulins against human tumor cell lines and murine B16F10-Nex2 melanoma aiming at the discovery of candidate molecules for cancer therapy. Four light-and heavy-chain CDR peptide sequences from different antibodies (C36-L1, HA9-H2, 1-H2 and Mg16-H2) showed cytotoxic activity against murine melanoma and a panel of human tumor cell lineages in vitro. Importantly, theyalso exerted anti-metastatic activity using a syngeneic melanoma model in mice. Other peptides (D07-H3, MN20v1, MS2-H3) were also protective against metastatic melanoma, without showing significant cytotoxicity against tumor cells in vitro. in this case, we suggest that these peptides may act as immune adjuvants in vivo. As observed, peptides induced nitric oxide production in bone-marrow macrophages showing that innate immune cells can also be modulated by these CDR peptides. the present screening supports the search in immunoglobulins of rather frequent CDR sequences that are endowed with specific antitumor properties and may be candidates to be developed as anti-cancer drugs. (C) 2014 Elsevier Inc. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Expt Oncol Unit UNONEX, São Paulo, SP, BrazilUniv Parma, Dept Biomed Biotechnol & Translat Sci, Microbiol & Virol Unit, I-43121 Parma, ItalyRecepta Biopharma, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Expt Oncol Unit UNONEX, São Paulo, SP, BrazilFAPESP: 2010/51423-0Web of Scienc

Serological analysis of dermatophyte isolates with monoclonal antibodies produced against Microsporum canis

Hybridoma cells produced by fusing myeloma cells with spleen cells from mice immunized with a soluble antigen of Microsporum canis yielded 30 antibody-producing clones. Six of these clones, propagated as ascites tumors in mice, showed two different types of monoclonal antibodies. The type 1 monoclonal antibody reacted with 17 heterologous and 10 homologous dermatophyte antigens. Type 2 monoclonal antibodies were unable to precipitate three antigens from different isolates of M. canis, thus suggesting the occurrence of different serotypes within the species

Therapeutic potential of yeast killer toxin-like antibodies and mimotopes

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Selection and characterization of a promoter for expression of single-copy recombinant genes in Gram-positive bacteria

BACKGROUND: In the past ten years there has been a growing interest in engineering Gram-positive bacteria for biotechnological applications, including vaccine delivery and production of recombinant proteins. Usually, bacteria are manipulated using plasmid expression vectors. The major limitation of this approach is due to the fact that recombinant plasmids are often lost from the bacterial culture upon removal of antibiotic selection. We have developed a genetic system based on suicide vectors on conjugative transposons allowing stable integration of recombinant DNA into the chromosome of transformable and non-transformable Gram-positive bacteria. RESULTS: The aim of this work was to select a strong chromosomal promoter from Streptococcus gordonii to improve this genetic system making it suitable for expression of single-copy recombinant genes. To achieve this task, a promoterless gene encoding a chloramphenicol acetyltransferase (cat), was randomly integrated into the S. gordonii chromosome and transformants were selected for chloramphenicol resistance. Three out of eighteen chloramphenicol resistant transformants selected exhibited 100% stability of the phenotype and only one of them, GP215, carried the cat gene integrated as a single copy. A DNA fragment of 600 base pairs exhibiting promoter activity was isolated from GP215 and sequenced. The 5' end of its corresponding mRNA was determined by primer extention analysis and the putative -10 and a -35 regions were identified. To study the possibility of using this promoter (PP) for single copy heterologous gene expression, we created transcriptional fusions of PP with genes encoding surface recombinant proteins in a vector capable of integrating into the conjugative transposon Tn916. Surface recombinant proteins whose expression was controlled by the PP promoter were detected in Tn916-containing strains of S. gordonii and Bacillus subtilis after single copy chromosomal integration of the recombinant insertion vectors into the resident Tn916. The surface recombinant protein synthesized under the control of PP was also detected in Enterococcus faecalis after conjugal transfer of a recombinant Tn916 containing the transcriptional fusion. CONCLUSION: We isolated and characterized a S. gordonii chromosomal promoter. We demonstrated that this promoter can be used to direct expression of heterologous genes in different Gram-positive bacteria, when integrated in a single copy into the chromosome

A novel microtubule de-stabilizing complementarity-determining region C36L1 peptide displays antitumor activity against melanoma in vitro and in vivo

Short peptide sequences from complementarity-determining regions (CDRs) of different immunoglobulins may exert anti-infective, immunomodulatory and antitumor activities regardless of the specificity of the original monoclonal antibody (mAb). In this sense, they resemble early molecules of innate immunity. C36L1 was identified as a bioactive light-chain CDR1 peptide by screening 19 conserved CDR sequences targeting murine B16F10-Nex2 melanoma. The 17-amino acid peptide is readily taken up by melanoma cells and acts on microtubules causing depolymerization, stress of the endoplasmic reticulum and intrinsic apoptosis. At low concentrations, C36L1 inhibited migration, invasion and proliferation of B16F10-Nex2 cells with cell cycle arrest at G2/M phase, by regulating the PI3K/Akt signaling axis involving Rho-GTPase and PTEN mediation. Peritumor injection of the peptide delayed growth of subcutaneously grafted melanoma cells. Intraperitoneal administration of C36L1 induced a significant immune-response dependent anti-tumor protection in a syngeneic metastatic melanoma model. Dendritic cells stimulated ex-vivo by the peptide and transferred to animals challenged with tumor cells were equally effective. The C36 VL CDR1 peptide is a promising microtubule-interacting drug that induces tumor cell death by apoptosis and inhibits metastases of highly aggressive melanoma cells

Aerosense: Long-Range Bluetooth Wireless Sensor Node for Aerodynamic Monitoring on Wind Turbine Blades

Predictive maintenance and structural health monitoring are challenging and promising research fields today. In particular, cost-effective and long-term monitoring of wind turbines has been proven to be one of the key elements to successfully increase their efficiency. Accurate numerical modeling and real-time control-in-the-loop play an increasingly prominent role in understanding and optimizing blade aerodynamic and acoustic performances. A non-intrusive and modular measurement system is a prerequisite for long-term measurement campaigns in existing and future wind turbines. Current methods of performing aerodynamic and acoustic field measurements are cumbersome and expensive, leading to a shortage of aerodynamic and acoustic datasets on operating wind turbines. This paper demonstrates the ability of the new Aerosense system to operate successfully in the field. Aerosense is a long-lasting battery-operated and flexible wireless sensor node that can directly measure aerodynamic and acoustic effects on wind turbine blades. It consists of an array of state-of-the-art Micro-Electro-Mechanical Systems (MEMS) sensors, including 40 barometers and 10 microphones, combined with an ultra low power system-on-chip with wireless transmission over Bluetooth 5.1. Experimental results demonstrate the possibility of continuously acquiring data for up to four months on a single lithium battery of 8.7 Ah, featuring an absolute accuracy of 10Pa and an audio bandwidth of 6kHz

Isolation of a Wickerhamomyces anomalus yeast strain from the sandfly Phlebotomus perniciosus, displaying the killer phenotype

The yeast Wickerhamomyces anomalus has been studied for its wide biotechnological potential, mainly for applications in the food industry. Different strains of W. anomalus have been isolated from diverse habitats and recently from insects, including mosquitoes of medical importance. This paper reports the isolation and phylogenetic characterization of W. anomalus from laboratory-reared adults and larvae of Phlebotomus perniciosus (Diptera: Psychodidae), a main phlebotomine vector of human and canine leishmaniasis. Of 65 yeast strains isolated from P. perniciosus, 15 strains were identified as W. anomalus; one of these was tested for the killer phenotype and demonstrated inhibitory activity against four yeast sensitive strains, as reported for mosquito-isolated strains. The association between P. perniciosus and W. anomalus deserves further investigation in order to explore the possibility that this yeast may exert inhibitory/killing activity against Leishmania spp