Shift in chicken intestinal gene association networks after infection with Salmonella

A primary infection of Salmonella enteritidis causes a spatial-temporal dependent change in the gene expression patterns in the intestine of chickens (Gallus gallus). This is the result of a dynamic intestinal response to adapt to the altered environment and to optimize its ‘health’ and functionality under the new circumstances. By inferring gene association networks (GANs), the complexities of and changes in biological networks can be uncovered. Within such GANs highly interacting (hub) genes can be identified, which are supposed to be high-level regulators connected to multiple processes. By exploring the intestinal expression of genes differing between control and Salmonella infected chicken in a time-dependent manner differences in GANs were found. In control chickens more developmental processes were observed, whereas in infected chickens relatively more processes were associated to ‘defense/pathogen response’. Moreover the conserved protein domains of the identified hub genes in controls were nuclear-associated, whereas hub genes in infected chickens were involved in ‘cellular communication’. The shift in topology and functionality of the intestinal GANs in control and Salmonella infected animals and the identification of GAN-specific hubs is a first step to understand the complexity of biological networks and processes regulating intestinal health and functionality under normal and disturbed conditions

FUNCTIONAL PHAGE DISPLAY OF CILIARY NEUROTROPHIC FACTOR

We report the display of human ciliary neurotrophic factor (hCNTF), a survival factor for neuronal cells belonging to the ~-helical cytokine superfamily, on the surface of the filamentous bacteriophage fd. The hCNTF cDNA was fused to a DNA sequence encoding the C-terminal domain of pIII, a minor coat protein exposed at one end of fd. Gene fusions were cloned into a plasmid containing the ColE1 plasmid and fd origins of replication, and were packaged into phagemid particles upon superinfection with M13KO7 helper phage. The resulting fusion phage bound specifically to anti-CNTF antibodies and to the recombinant soluble CNTF s-receptor. Moreover, phage-displayed hCNTF was found to possess biological activity at concentrations comparable to those of the soluble cytokine. These results demonstrate that CNTF can be displayed on phage in a correctly folded and functionally active form. Binding of fusion phage to immobilized CNTF ~-receptor and subsequent elution at low pH resulted in affinity purification of CNTF-displaying virions. Utilization of this technology should enable the selection of high-affinity variants from libraries of CNTF mutants displayed on phage

L’approccio read-through per il trattamento della fibrosi cistica causata da mutazioni di stop Durata: 24 mesi (dal 01/09/2012 al 31/08/2014) Progetto n°: 01/2012 FFC Finanziato da: Fondazione per la Ricerca sulla Fibrosi Cistica-Bando 2012

Durata: 24 mesi (dal 01/09/2012 al 31/08/2014) Progetto n°: 01/2012 FFC Finanziato da: Fondazione per la Ricerca sulla Fibrosi Cistica-Bando 2012-Nonsense mutations are the leading cause of approximately 30% of inherited diseases, including cystic fibrosis (CF). They promote premature translational termination and following loss of CFTR protein by introduction of premature termination codons (PTCs). In the last few years, it has been demonstrated that drugs (like aminoglycoside antibiotics) can be designed and produced to suppress this process by the ribosomal read-through mechanism where these drugs mask PTC synthetizing a full-lengh CFTR protein. The rationale supporting of this project is to optimize the ribosomal read-through molecules leading to restoration of CFTR production in cystic fibrosis caused by stop mutation. This study may introduce new hopes for the development of a pharmacologic approach to the cure of CF

The Tanala, a hill tribe of Madagascar,

v.22 (1933

Efficacy of PEGylated ciliary neurotrophic factor superagonist variant in diet-induced obesity mice

: Ciliary neurotrophic factor (CNTF) is a neurotrophic cytokine able to induce appetite reduction, weight loss and antidiabetic effects. However, its susceptibility to neutralizing anti-CNTF antibodies in patients hampered its use for treatment of human obesity and diabetes. In addition, CNTF has a very short plasma half-life, which limits its use as a therapeutic agent. Solutions, directed to prolong its in vivo effects, vary from the implantation of encapsulated secreting cells to identification of more active variants or chemical modification of the protein itself. PEGylation is a widely used modification for shielding proteins from circulating antibodies and for increasing their plasma half-life. Here, we have selected DH-CNTF, a CNTF variant which has a 40-fold higher affinity for the CNTF receptor α accompanied by an increased activity in cellular assays. The PEGylated DH-CNTF retained the biological activity of native protein in vitro and showed a significant improvement of pharmacokinetic parameters. In an acute model of glucose tolerance, the PEG-DH-CNTF was able to reduce the glycemia in diet-induced obese animals, with a performance equaled by a 10-fold higher dose of DH-CNTF. In addition, the PEGylated DH-CNTF analog demonstrated a more potent weight loss effect than the unmodified protein, opening to the use of CNTF as weight reducing agent with treatment regimens that can better meet patient compliance thanks to reduced dosing schedules