Mutational spectrum of F8 gene and prothrombotic gene variants in haemophilia A patients from Southern Italy.

Haemophilia A (HA) is an X-linked recessive haemorrhagic disorder caused by a deficiency of coagulation factor VIII. Disease causative mutations are heterogeneous and spread all over the F8 gene sequence, with the exception of intron 22 inversion occurring in about 50% of severe patients. To define the specific mutational repertoire and mutation detection rate, we analysed F8 gene, by polymerase chain reaction and direct sequencing, in 128 unrelated patients from Southern Italy with severe (n = 108), moderate (n = 9) or mild (n = 11) HA. We identified 120 mutations, including 64 cases of intron 22 inversion (53.3%), three of intron 1 inversion (2.5%), one large deletion (0,8%) and 52 point mutations (43.3%). In particular, 19/120 were novel and 7/52 point mutations (13.5%) occurred at CpG sites. We also investigated eight prothrombotic genetic variants in a subgroup of 74 severe HA patients to evaluate their possible protective effect on the severity of clinical expression. Methylenetetrahydrofolate reductase A1298C and plasminogen activator inhibitor-1 4G variants recurred more frequently in HA patients with a less-severe phenotype. Clinical impact of these findings needs large-scale studies to further define the role of these prothrombotic variants as possible modifier factors of HA phenotype.

Butyrate modulating effects on pro-inflammatory pathways in human intestinal epithelial cells.

Butyrate acts as energy source for intestinal epithelial cells and as key mediator of several immune processes, modulating gene expression mainly through histone deacetylation inhibition. Thanks to these effects, butyrate has been proposed for the treatment of many intestinal diseases. Aim of this study was to investigate the effect of butyrate on the expression of a large series of target genes encoding proteins involved in pro-inflammatory pathways. We performed quantitative real-time-PCR analysis of the expression of 86 genes encoding proteins bearing to pro-inflammatory pathways, before and after butyrate exposure, in primary epithelial cells derived from human small intestine and colon. Butyrate significantly down-regulated the expression of genes involved in inflammatory response, among which nuclear factor kappa beta, interferon-gamma, Toll like 2 receptor and tumour necrosis factor-alpha. Further confirmations of these data, including studies at protein level, would support the use of butyrate as effective therapeutic strategy in intestinal inflammatory disorders.

Pseudouridine and 1-ribosylpyridin-4-one-3-carboxamide (PCNR) serum concentrations in human immunodeficiency virus type 1-infected patients are independent predictors for AIDS progression.

A prospective study was conducted with 161 human immunodeficiency virus (HIV)-positive patients to investigate the prognostic role of 10 serum-modified nucleosides with regard to some of the most widely used parameters of AIDS progression. Serum concentrations of pseudouridine (> 3.77 nmol/mL) predicted progression to AIDS in CDC stage A2 HIV-infected patients much better than did other widely used parameters (hazard ratio, 2.7; 95% confidence interval, 1.29-6.35; P = .01; median permanence time in stage A2, 17 vs. 30.5 months; P = .03). Serum concentrations of 1-ribosylpyridin-4-one-3-carboxamide (PCNR) and beta 2-microglobulin and the CD4:CD8 cell ratio, in decreasing order and used in combination, differentiated the overall survival time probability of AIDS patients; PCNR was the best and a new independent predictor (overall survival time, > 31 months, no positive parameters; 19.3 months, one positive parameter; and 5.5 months, two positive parameters.