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2.
AMB Express ; 1(1): 21, 2011 07 19.
Article in English | MEDLINE | ID: mdl-21906377

ABSTRACT

Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria, making it difficult to assess survival under different conditions. Within this context, the principal aims of this work were: (1) to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds; (2) to establish the most representative mathematical function for B. japonicum mortality on soybean seeds after inoculation; (3) to evaluate if environmental or physiological conditions modify B. japonicum mortality on soybean seeds; and (4) to create a new protocol for quality control of soybean inoculants. We successfully evaluated the combination of pentachloronitrobenzene and vancomycin added to the yeast-mannitol medium to inhibit most fungi and Gram-positive soybean microbiota, thus producing reliable counts of B. japonicum from inoculated soybean seeds. Percentages of recovery and survival factors were obtained and used to construct a two-phase exponential decay non-linear regression function. High temperature and desiccation decreased these parameters, while the optimization of temperature and the use of osmoprotective compounds with inoculants increased them. The use of this protocol minimized heterogeneity between experiments and may be considered more reliable than the simple expression of direct colony count of bacteria recovered from seeds.

3.
Cell Mol Life Sci ; 68(14): 2453-67, 2011 Jul.
Article in English | MEDLINE | ID: mdl-20972812

ABSTRACT

The receptor (CXCR4) for the stromal-derived factor-1 (SDF1) and the urokinase-receptor (uPAR) are up-regulated in various tumors. We show that CXCR4-transfected cells migrate toward SDF1 on collagen (CG) and do not on vitronectin (VN). Co-expression of cell-surface uPAR, which is a VN receptor, impairs SDF1-induced migration on CG and allows migration on VN. Blocking fMLP receptors (fMLP-R), alpha-v integrins or the uPAR region capable to interact with fMLP-Rs, impairs migration of uPAR/CXCR4-transfected cells on VN and restores their migration on CG. uPAR co-expression also reduces the adherence of CXCR4-expressing cells to various components of the extracellular matrix (ECM) and influences the partitioning of beta1 and alpha-v integrins to membrane lipid-rafts, affecting ECM-dependent signaling. uPAR interference in CXCR4 activity has been confirmed in cells from prostate carcinoma. Our results demonstrate that uPAR expression regulates the adhesive and migratory ability of CXCR4-expressing cells through a mechanism involving fMLP receptors and alpha-v integrins.


Subject(s)
Receptor Cross-Talk , Receptors, CXCR4/metabolism , Receptors, Formyl Peptide/metabolism , Receptors, Urokinase Plasminogen Activator/metabolism , Blotting, Western , Cell Adhesion , Cell Line, Tumor , Cell Membrane/metabolism , Cell Movement , Chemokine CXCL12/metabolism , Collagen/metabolism , Enzyme Activation , HEK293 Cells , Humans , Integrin alpha5beta1/metabolism , Membrane Microdomains/metabolism , Microscopy, Confocal , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , RNA Interference , Receptors, CXCR4/genetics , Receptors, Formyl Peptide/genetics , Receptors, Urokinase Plasminogen Activator/genetics , Transfection , Vitronectin/metabolism
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