Diabetes mellitus was defined based on World Health Organization criteria

arameter estimates for each run showed ESS values >250. The trees were summarized in a target tree by the Tree Annotator program included in the BEAST package by choosing the tree with the maximum product of posterior probabilities after a 50% burn-in. The tree was Nigericin (sodium salt) rooted using a midpoint rooting by Fig-Tree software version 1.4.2. Resistance Analysis and Tropism Prediction HIV-1 strains were defined as resistant if carrying at least one drug resistance mutation among the mutations listed by Bennett et al. PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19725016 and the primary mutations reported in the IAS-USA list and/or the HIV Drug Resistance Stanford Database. Polymorphisms at positions already associated with drug resistance were also investigated. HIV-1 co-receptor usage was inferred from the V3 nucleotide sequence by using the Geno2Pheno algorithm available at the following website: http://coreceptor.bioinf.mpi-inf.mpg.de/. 4 / 17 HIV-1 Transmission Clusters in Newly Diagnosed Men The analysis was performed setting Geno2Pheno at false positive rate of 10%. Thus, sequences with FPR <10% were considered X4/DM tropic. Statistical Analyses Differences regarding the epidemiological characteristics between the two clusters were evaluated as follows: i) for the categorical PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19723728 variables, by Fisher’s exact test ii) for the continuous variables, by the Kruskal-Wallis test. In all the analyses performed, P values <0.05 were considered as statistically significant. The statistical program used was SPSS for Windows. Results Clusters Identification Among 1,546 new cases of HIV-1 infections diagnosed between May 2011 and September 2014 in different counselling and testing centres in Central Italy, 534 belonged to non-B subtypes. Demographics and viro-immunological characteristics of these 534 patients were reported in. Among the 534 HIV-1 non-B infected patients, 35 drew our attention, because by a preliminary phylogenetic analysis by the NJ method, performed on 2,158 pol non-B subtype sequences obtained for routine clinical practice between January 2000 and December 2014, their HIV-1 pol sequences formed two distinct clusters. The statistical robustness of both clusters was confirmed by the ML and the Bayesian phylogenetic trees. These trees were performed using 75 C and 10 and CRF17_BF pol sequences, including the 35 sequences identified in the preliminary phylogenetic analysis and 50 pol sequences randomly selected from the 2,158 obtained for routine clinical practice. Both clusters were characterized by a bootstrap value of 100%, and a posterior probability of one. An additional confirmation of the high homology among sequences involved in the two clusters arose from the extremely low mean genetic distance observed for the pol gene in both C and CRF17_BF clusters. The results obtained by the phylogenetic analyses on the pol gene were also confirmed by using the V3 loop, despite the short length of the region and its higher variability compared to pol gene . Clusters Dating HIV-1 Transmission Clusters in Newly Diagnosed Men Fig 2. Maximum likelihood tree of the 10 CRF17_BF sequences plus the 3 CRF17_BF references. The ML tree was inferred by using PhyML program. Transmission clusters were identified by a bootstrap support >90%. The tree was rooted using a midpoint rooting. Bayesian phylogenetic tree of the 10 CRF17_BF sequences plus the 3 CRF17_BF references. The Bayesian phylogenetic tree was inferred by using MrBayes. Clades with a posterior probability of one were considered epidemiological clust