ABSTRACT
BACKGROUND: The scorpion Tityus stigmurus is widely distributed in Northeastern Brazil and known to cause severe human envenoming, inducing pain, hyposthesia, edema, erythema, paresthesia, headaches and vomiting. The present study uses a transcriptomic approach to characterize the gene expression profile from the non-stimulated venom gland of Tityus stigmurus scorpion. RESULTS: A cDNA library was constructed and 540 clones were sequenced and grouped into 153 clusters, with one or more ESTs (expressed sequence tags). Forty-one percent of ESTs belong to recognized toxin-coding sequences, with transcripts encoding antimicrobial toxins (AMP-like) being the most abundant, followed by alfa KTx- like, beta KTx-like, beta NaTx-like and alfa NaTx-like. Our analysis indicated that 34% of the transcripts encode "other possible venom molecules", which correspond to anionic peptides, hypothetical secreted peptides, metalloproteinases, cystein-rich peptides and lectins. Fifteen percent of ESTs are similar to cellular transcripts. Sequences without good matches corresponded to 11%. CONCLUSIONS: This investigation provides the first global view of gene expression of the venom gland from Tityus stigmurus under resting conditions. This approach enables characterization of a large number of venom gland component molecules, which belong either to known or non yet described types of venom peptides and proteins from the Buthidae family.
Subject(s)
Gene Expression Profiling , Scorpion Venoms/genetics , Scorpions/metabolism , Amino Acid Sequence , Animals , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/metabolism , Expressed Sequence Tags , Gene Library , Insulin-Like Growth Factor Binding Proteins/chemistry , Insulin-Like Growth Factor Binding Proteins/metabolism , Lectins/chemistry , Lectins/metabolism , Molecular Sequence Data , Neurotoxins/chemistry , Neurotoxins/metabolism , Peptides/chemistry , Peptides/metabolism , Scorpion Venoms/classification , Scorpion Venoms/metabolism , Sequence AlignmentABSTRACT
AIMS: To conduct a morphological, functional and chromosomal characterization of mesenchymal stem cell populations from the human subendothelium umbilical cord vein after cryopreservation. MATERIAL & METHODS: Five human umbilical cords were processed in order to obtain mesenchymal stem cells. Flow cytometry, differentiation assays and cytogenetic analysis were carried out before and after the cryopreservation process. RESULTS: Flow cytometry revealed that CD105, CD73 and CD90 markers were expressed by the cells, which lacked the expression of hematopoietic lineage markers, such as CD14, CD34 and CD45. The mesenchymal stem cells demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation. Chromosome analysis showed no clonal chromosome changes in the cells in either situation. However, a significant number of nonclonal chromosomal aberrations were apparent after cryopreservation, including monosomies and structural changes. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration. This was present both before and after experimental procedures. CONCLUSION: These findings show that using mesenchymal stem cells for clinical approaches requires careful investigation and sensitive tests in order to ensure cellular therapy biosafety.
Subject(s)
Chromosomes, Human/metabolism , Cryopreservation/methods , Endothelium/cytology , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Umbilical Cord/blood supply , Umbilical Veins/cytology , Antigens, Surface/metabolism , Biomarkers/metabolism , Cell Differentiation , Cell Separation , Cell Shape , Chromosome Aberrations , Endothelium/metabolism , Female , Flow Cytometry , Humans , KaryotypingABSTRACT
UNLABELLED: This article reports the genotoxicity assessment of an extract of M. oleifera seed powder and the water-soluble Moringa oleifera lectin (WSMoL) isolated from seeds. The lectin isolated by chitin chromatography showed hemagglutinating activity with different erythrocytes, activity in a broad pH range (4.5 to 9.5), and retention of hemagglutinating activity after being heated to 100 °C. Genotoxicity of the seed extract and WSMoL were assessed using the cell-free plasmid DNA as well as the Salmonella typhimurium (Ames and Kado) assays with TA97, TA98, TA100, and TA102 in the presence or absence of hepatic metabolization. Seed extract at concentration (0.2 µg/µL) recommended to treat water was not genotoxic by Ames, Kado, and cell-free plasmid DNA assays. S. typhimurium strains showed to be sensitive to M. oleifera extract revealing a mutagenic effect at doses higher than 0.6 µg/µL with hepatic metabolization. The extract at doses higher than 0.4 µg/µL, without hepatic metabolization, was mutagenic for TA100 and TA102. WSMoL was nonmutagenic by used assays. The use of high concentrations of the extract may pose a risk to human health and the safe use of M. oleifera seed powder to treat water for human consumption requires more study; however, the purified lectin could be an alternative for water treatment. PRACTICAL APPLICATION: The concentration 0.2 µg/µL of M. oleifera seed extract recommended to treat water for humans did not pose a risk to human health. The mutagenicity detected at concentrations higher than 0.4 µg/µL was not due to WSMoL, lectin isolated from extract.
