These results demonstrate the potential utility of selected biomarkers to distinguish patients with the highest risk for treatment failure and bacteremia-related complications, providing a valuable tool for clinicians in the management of S. aureus bacteremia. Additionally, these biomarkers could identify patients with the greatest potential to benefit from novel therapies in clinical trials.

Neural progenitor cells were isolated from rat fetal telencephalon and proliferate as neurospheres in the presence of EGF, FGF-2, and heparin. In the absence of these growth factors, neurospheres differentiate into neurons, astrocytes, and oligodendrocytes. Using an embryonal carcinoma cell line as in vitro differentiation model, we have already demonstrated the presence of an autocrine loop system between kinin-B2 receptor activity and secretion of its ligand bradykinin (BK) as prerequisites for final neuronal differentiation (Martins et al., J Biol Chem 2005; 280: 19576-19586). The aim of this study was to verify the activity of the kallikrein-kinin system (KKS) during neural progenitor cell differentiation. Immunofluorescence studies and flow cytometry analysis revealed increases in glial fibrillary acidic protein and b-3 tubulin expression and decrease in the number of nestin-positive cells along neurospheres differentiation, indicating the transition of neural progenitor cells to astrocytes and neurons. Kinin-B2 receptor expression and activity, secretion of BK into the medium, and presence of highmolecular weight kininogen suggest the participation of the KKS in neurosphere differentiation. Functional kinin-B2 receptors and BK secretion indicate an autocrine loop during neurosphere differentiation to neurons, astrocytes, and oligodendrocytes, reflecting events occurring during early brain development. ' 2008 International Society for Analytical CytologyKey terms kinin-B2 receptor; neural differentiation; neurosphere; kallikrein-kinin system BRADYKININ (BK), kallidin, des-Arg 9 -BK, and des-Arg 9 -kallidin are the biological active peptides of the kallikrein-kinin system (KKS). BK and kallidin as ligands of Gprotein-coupled kinin-B2 receptors (B2BKR) are generated upon proteolytic cleavage of high-or low-molecular weight kininogen (HMWK or LMWK) by plasma-or tissue-kallikrein serine protease, respectively. Carboxy-terminal arginines are removed from these biological active peptides by carboxypeptidases M or N to originate the kinin-B1 receptor (B1BKR) agonists des-Arg 9 -BK and des-Arg 9 -kallidin (Fig. 1). Stimulation of the B2BKR by its agonists results in the activation of phospholipase C-b (PLC-b), generating diacyl glycerol and inositol 1,4,5-triphosphate (IP 3 ) and resulting in release of Ca 21 from intracellular IP 3 -sensitive stores. Furthermore, BK mediates the activation of endothelial nitric oxide synthase (1) and stimulates the phospholipase A2 activity (2). B2BKR also activates proteins with tyrosine kinase activity (3), and the receptor can directly interact with neuronal and endothelial nitric oxide synthetase (nNOS and eNOS), resulting in NO production (4). Both B1BKR and B2BKR are coupled to G aq and G ai proteins (5,6) and triggered the same signaling pathways, but differ in their expression pattern and intensities of receptorinduced calcium responses and receptor-desensitization rates (7).B2BKR is constitutively expressed and broadly distributed throughout the tissues, and B1BK...

Cystic fibrosis (CF) is a recessive inherited disease associated with multiorgan damage that compromises epithelial and inflammatory cell function. Induced pluripotent stem cells (iPSCs) have significantly advanced the potential of developing a personalized cell-based therapy for diseases like CF by generating patient-specific stem cells that can be differentiated into cells that repair tissues damaged by disease pathology. The F508del mutation in airway epithelial cell-derived CF-iPSCs was corrected with small/short DNA fragments (SDFs) and sequence-specific TALENs. An allele-specific PCR, cyclic enrichment strategy gave ~100-fold enrichment of the corrected CF-iPSCs after six enrichment cycles that facilitated isolation of corrected clones. The seamless SDF-based gene modification strategy used to correct the CF-iPSCs resulted in pluripotent cells that, when differentiated into endoderm/airway-like epithelial cells showed wild-type (wt) airway epithelial cell cAMP-dependent Cl ion transport or showed the appropriate cell-type characteristics when differentiated along mesoderm/hematopoietic inflammatory cell lineage pathways.

Background and purpose: Kinins are implicated in many pathophysiological conditions, and recent evidence has suggested their involvement in colitis. This study assessed the role of the kinin B 1 receptors in a mouse model of colitis. Experimental approach: Colitis was induced in mice by 2,4,6-trinitrobenzene sulphonic acid (TNBS), and tissue damage and myeloperoxidase activity were assessed. B 1 receptor induction was analysed by organ bath studies, binding assay and reverse transcription PCR. Key results: TNBS-induced colitis was associated with tissue damage, neutrophil infiltration and time-dependent increase of colon B 1 receptor-mediated contraction, with the maximal response observed at 72 h. The upregulation of the B 1 receptor at this time point was also confirmed by means of binding studies. B 1 receptor mRNA levels were elevated as early as 6 h after colitis induction and remained high for up to 48 h. TNBS-evoked tissue damage and neutrophil influx were reduced by the selective B 1 receptor antagonist SSR240612, and in B 1 receptor knockout mice. In vivo treatment with inhibitors of protein synthesis, nuclear factor-kB activation, inducible nitric oxide synthase (iNOS) or tumour necrosis factor a (TNFa) significantly reduced B 1 receptor agonist-induced contraction. Similar results were observed in iNOS and TNF receptor 1-knockout mice. Conclusions and implications: These results provide convincing evidence on the role of B 1 receptors in the pathogenesis of colitis. Therefore, the blockade of kinin B 1 receptors might represent a new therapeutic option for treating inflammatory bowel diseases.

Thimet oligopeptidase (THOP1) is thought to be involved in neuropeptide metabolism, antigen presentation, neurodegeneration, and cancer. Herein, the generation of THOP1 C57BL/6 knockout mice (THOP1−/−) is described showing that they are viable, have estrus cycle, fertility, and a number of puppies per litter similar to C57BL/6 wild type mice (WT). In specific brain regions, THOP1-/- exhibit altered mRNA expression of proteasome beta5, serotonin 5HT2a receptor and dopamine D2 receptor, but not of neurolysin (NLN). Peptidomic analysis identifies differences in intracellular peptide ratios between THOP1-/- and WT mice, which may affect normal cellular functioning. In an experimental model of multiple sclerosis THOP1-/- mice present worse clinical behavior scores compared to WT mice, corroborating its possible involvement in neurodegenerative diseases. THOP1-/- mice also exhibit better survival and improved behavior in a sepsis model, but also a greater peripheral pain sensitivity measured in the hot plate test after bradykinin administration in the paw. THOP1-/- mice show depressive-like behavior, as well as attention and memory retention deficits. Altogether, these results reveal a role of THOP1 on specific behaviors, immune-stimulated neurodegeneration, and infection-induced inflammation.