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Serotyping was performed by capsular swelling (Quellung) reaction using commercial serogroup and serotype-specific antiserum at the Spanish Reference Laboratory (Instituto Carlos III, Madrid), where all isolates were routinely sent. Serotypes were classified into two groups: PCV7 serotypes (serotypes Selinexor in vivo 4, 6B, 9V, 14, 18C, 19F and 23F) and non-PCV7 serotypes (all others). Information regarding serotype is lacking in 13.5% of the isolates, in most cases because samples were not sent to the reference centre. Only 1% of cases were non-typeable pneumococci. We calculated overall and group age incidence rates of IPP by 100?000 persons-year. In order to make these calculations we used as denominator the exact number of the population living in the reference area of the hospital per year according to the Department of Statistics in Catalunya [19]. Hospital Vall d��Hebron is the only tertiary hospital in the reference area and the contribution of other hospitals in the area is minimal and has not changed during the study period. In order to avoid some bias related to the reference population, we also calculated selleck the incidence of IPP per 1000 adult admissions in our hospital per year. During the study period the overall number of admissions per 100?000 of the catchment population has been constant. To estimate the incidence of IPP due to vaccine or non-vaccine serotypes, we assumed that the distribution of serotypes for cases missing serotype information was the same as the distribution for those cases with serotype information. Statistical analyses were performed using the statistical software package SPSS for Windows, version 15.0. Differences in means of the incidence were tested using the Mantel�CHaenszel test [20]. The absolute risk difference with corresponding 95% confidence intervals were reported. 3-mercaptopyruvate sulfurtransferase The chi-square test was used to compare the distribution of categorical variables and the Student��s t-test for continuous variables. To asses differences in disease characteristics between the two periods we compared overall and age group patients. A univariable and multivariable logistic regression analysis was performed to identify variables independently associated with death and septic shock. Results were considered statistically significant if the two-tailed p value was