1, Table 1). We directly examined 7 patients belonged to IV (IV-5, IV-8, IV-9, IV-11) and III generations (III-3, III-6, III-8), whilst the remaining 17 patients were identified from medical records. Certainly no generation was skipped from the second, suggesting an autosomal dominant inheritance. Table 1. Detailed patient characteristics. Methods After obtaining written Cyclosporin A chemical structure informed consent, genomic DNA from the proband and the affected members IV-8 and IV-11 was extracted from leukocytes of whole blood samples. The remaining patients of the family did not give their consent or were not available for the analysis. The five coding

exons of SOD1 gene and at least 30 bp of flanking intronic sequence, Inhibitors,research,lifescience,medical amplified by polymerase chain reaction (PCR), were sequenced using the Big- Dye Terminator Cycle Sequencing Ready Inhibitors,research,lifescience,medical Reaction Kit (Applied Biosystems) and run on a capillary sequencer (ABI Prism 310 Genetic Analyzer, Applied Biosystems). TARDBP, FUS/TLS and C9ORF72 genes were also screened to better characterize the genotypes of the three patients. Results DNA analysis of the proband and members IV-8 and IV-11 showed a heterozygous mutation c.149T>C in the exon 5 of the SOD1 gene, causing a substitution of isoleucine to threonine threonine (p.Ile149Thr). Regarding TARDBP, FUS/TLS and C9ORF72, the Inhibitors,research,lifescience,medical three

patients showed no pathologic mutations. Discussion We report the first Inhibitors,research,lifescience,medical Italian kindred of FALS due to exon 5 missense mutation c.149T>C in the SOD1 gene. Previously, the same mutation has been identified in a few Caucasian (7, 8) and Asian (9) families, and has been

revealed able of inducing structural modifications of the relative charges of amino acids, significantly affecting the SOD1 enzymatic activity. In fact, about p.Ile149Thr mutation, it was found that the heterodimers composed Inhibitors,research,lifescience,medical by one normal and one mutant molecules appeared to be less efficient or stable, causing a relevant destabilization of SOD1 dimer structure, and promoting the accumulation of toxic intracellular aggregates Adenylyl cyclase (7, 8). However, the exact mechanisms by which mutant SOD1 (mSOD1) causes motor neuronal cell death have yet to be established (1). Recently, evidence from transgenic models expressing mSOD1 has allowed to hypothesize a potential contribution of non-motor neuron cells, such as microglia, in triggering an alteration of the balance between neuroprotection and cytotoxicity in favor of the latter (10). In fact, misfolded proteins, such as mSOD1, seem to induce impairment of mitochondrial function and axoplasmic flow and release from motor neurons of abnormal signals able to activate microglia. About genotype-phenotype correlations, in our case, the clinical presentation of the seven patients examined (III-3, III-6, III-8, IV-5, IV-8, IV-9, IV-11) was characterized by mean age of onset of 40.8 ± 9.