Unconventional epitopes presented by HLA class I complexes are emerging targets for T cell targeted immunotherapies. Their identification by mass spectrometry (MS) required development of novel methods to cope with the large number of theoretical candidates. Methods to identify post‐translationally spliced peptides led to a broad range of outcomes. We here investigated the impact of three common database search engines – that is, Mascot, Mascot+Percolator, and PEAKS DB – as final identification step, as well as the features of target database on the ability to correctly identify non‐spliced and cis‐spliced peptides. We used ground truth datasets measured by MS to benchmark methods’ performance and extended the analysis to HLA class I immunopeptidomes. PEAKS DB showed better precision and recall of cis‐spliced peptides and larger number of identified peptides in HLA class I immunopeptidomes than the other search engine strategies. The better performance of PEAKS DB appears to result from better discrimination between target and decoy hits and hence a more robust FDR estimation, and seems independent to peptide and spectrum features here investigated.

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Purpose: to investigate the influence of the training process of high-class and elite sprint swimmers of both genders on the parameters of the autonomic nervous system, central hemodynamics and physical working capacity. Material & Methods: in the preparatory period of the training process, 145 sprint swimmers (56 females and 89 males) with sports qualifications from CMS (high-class swimmers) to MS-MSIC (elite swimmers) were examined, of which 56 sprint swimmers (31 females and 25 males) in 2006 and 89 sprint swimmers in 2021 (25 females and 64 males). To assess the autonomic regulation of cardiac activity time- and frequency domain methods of HRV analysis were used. Central hemodynamic parameters were examined by an automated tetrapolar rheography. Physical working capacity was measured according to a common technique on a cycling ergometer using a submaximal PWC170 test and calculating the relative value of physical working capacity, i.e. PWC170/kg. Results: comparison of the studied parameters in sprint swimmers examined in 2006 and 2021 showed a significant decrease in PWC170/kg in present-day high-class sprint swimmers with sports qualification CMS (both females and males), by 23.84%, (p=0.0003) and 12.68% (p=0.018), respectively, and in elite male swimmers with sports qualifications MS-MSIC by 9.46% (p=0.017), as well as a downward trend in elite female swimmers with sports qualifications MS-MSIC by 18.28% (p=0.154). Differences in HRV and central hemodynamics parameters were statistically insignificant. Conclusions: in present-day high-class female swimmers, the predominance of the sympathetic arm of the ANS was revealed, the value of PWC170/kg was 23.84% (p=0.0003) lower than in female swimmers of the same level in 2006. There were no significant differences between all the studied indices of elite female swimmers in 2021 and the indices of female swimmers of the same level in 2006. In present-day elite male swimmers and high-class male swimmers, the value of PWC170/kg was lower, respectively, by 9.46% (p=0.017) and 12.68% (p=0.018) than in swimmers of the same level in 2006. A significant decrease in the functional state of modern swimmers-sprinters of both genders against an earlier achievement of qualification standards than their peers in 2006 may be due to a change in the modern methodology of the training process aimed at reducing the training load in the zone of development of general endurance (aerobic orientation) by increasing the load of speed and power (anaerobic) orientation.