BUSCADOR RECOLECTA

Supplemental Table S1. Complete overview of the inclusion and exclusion criteria for this study. Supplemental Table S2. Composition of the EuroFlow B-cell panel and technical information on the reagents for the IMI-2 PERISCOPE BERT study. Supplemental Table S3. Phenotypic descriptions used to define B-cell subsets stained with the EuroFlow B-cell panel by manual analysis. Supplemental Table S4. Baseline distribution of leukocytes, lymphocytes, T cells, and NK cells in donor groups. Supplemental Table S5. Spearman Ranking Correlation between IgG1+ plasma cell and memory B-cell kinetics and vaccine-component-specific serum IgG. Supplemental Table S6. Spearman Ranking Correlation between IgA1+ plasma cell and IgA memory B-cell kinetics and vaccine-component-specific serum IgA. Supplemental Figure S1. No clear over-time postvaccination changes in major populations in any of the donor groups. Supplemental Figure S2. Over-time maturation of total plasma cells. Supplemental Figure S3. No significant changes in IgG1+ memory B-cell subsets upon vaccination. Supplemental Figure S4. Correlation between cellular changes as measured by flow cytometry and ELISpot. Supplemental Figure S5. Correlation between cellular changes and the vaccine-specific serum IgG level postvaccination as determined by Spearman’s Ranking Correlation per age cohort. Supplemental Figure S6. Impact of sex on cellular responses after vaccination in the young adult cohort (all wP-primed). Supplemental Figure S7. IgG1+ and total plasma cell expansion is more prominent in non-age-matched donors after wP priming., Peer reviewed

The following are available online at https://www.mdpi.com/article/10.3390/cancers14020449/s1. Supplementary Figures. Figure S1: Distribution of pathological CSF samples (without healthy ones) among each phase of study and the different groups according to the incidence of the pathology, depending on the infiltration (CSF +/− LM) and the primary tumor (hematologic and solid tumor). Figure S2: Quality control images of the planar protein microarrays generated. Figure S3: A quantile normalization in planar protein microarrays. Figure S4: Coomasie gels which indicate protein distribution across samples. Figure S5: Venn diagrams of total identified proteins with LC-MS/MS. Figure S6: Plots showing the functional proteins using the Reactome for different conditions. Figure S7: Differential protein profiles within CSF + LM according to primary tumor (Lymphoma) by protein microarrays. Figure S8: Differential protein profiles within CSF + LM according to primary tumor (Leukemia) by protein microarrays. Figure S9: Differential protein profiles within CSF + LM according to primary tumor (Lymphoma) by affinity proteomics. Figure S10: Differential protein profiles within CSF + LM according to primary tumor (Leukemia) by affinity proteomics. Figure S11: Summary of the multipronged proteomics characterization among the different phases of study. Supplementary Tables. Table S1: Table of clinical-biological characteristics from the whole CSF samples used in the study. Table S2: Antibodies list used in Planar Protein Microarrays. Table S3: Antibodies list used in Beads Suspension Microarrays. Table S4: Protein identification with LC-MS/MS among the different strategies and their emPAI quantification. Table S5: Boxplots of the protein identified in validation and confirmation phases, respectively, comparing the different groups of study. Table S6: Intensity data results from Planar Protein Microarrays. Table S7: Intensity data results from Beads Suspension Microarrays. Table S8: ROC analysis list of potential biomarker panel on CSF +/− LM and the different comparisons by protein arrays and affinity proteomics., Peer reviewed

Innate myeloid cell (IMC) populations form an essential part of innate immunity. Flow cytometric (FCM) monitoring of IMCs in peripheral blood (PB) has great clinical potential for disease monitoring due to their role in maintenance of tissue homeostasis and ability to sense micro-environmental changes, such as inflammatory processes and tissue damage. However, the lack of standardized and validated approaches has hampered broad clinical implementation. For accurate identification and separation of IMC populations, 62 antibodies against 44 different proteins were evaluated. In multiple rounds of EuroFlow-based design-testing-evaluation-redesign, finally 16 antibodies were selected for their non-redundancy and separation power. Accordingly, two antibody combinations were designed for fast, sensitive, and reproducible FCM monitoring of IMC populations in PB in clinical settings (11-color; 13 antibodies) and translational research (14-color; 16 antibodies). Performance of pre-analytical and analytical variables among different instruments, together with optimized post-analytical data analysis and reference values were assessed. Overall, 265 blood samples were used for design and validation of the antibody combinations and in vitro functional assays, as well as for assessing the impact of sample preparation procedures and conditions. The two (11- and 14-color) antibody combinations allowed for robust and sensitive detection of 19 and 23 IMC populations, respectively. Highly reproducible identification and enumeration of IMC populations was achieved, independently of anticoagulant, type of FCM instrument and center, particularly when database/software-guided automated (vs. manual “expert-based”) gating was used. Whereas no significant changes were observed in identification of IMC populations for up to 24h delayed sample processing, a significant impact was observed in their absolute counts after >12h delay. Therefore, accurate identification and quantitation of IMC populations requires sample processing on the same day. Significantly different counts were observed in PB for multiple IMC populations according to age and sex. Consequently, PB samples from 116 healthy donors (8-69 years) were used for collecting age and sex related reference values for all IMC populations. In summary, the two antibody combinations and FCM approach allow for rapid, standardized, automated and reproducible identification of 19 and 23 IMC populations in PB, suited for monitoring of innate immune responses in clinical and translational research settings., Peer reviewed

Supplemental Table 1. Detailed biological and demographic features of patients. Supplemental Table 2: Composition of the EuroFlow B-CLPD panel Supplemental Table 3. Overview on the data analysis strategy within the scope of the main study Supplemental Table 4. Canonical coefficients for CA1 and CA2. Significance of contribution of individual parameters to the canonical axes CA1 and CA2 by differential diagnosis. V Supplemental Table 5. SD lines utilized as decision criterion per pair-wise differential diagnosis Supplemental Table 6. Medians (10th – 90th 309 percentile) of medFIs and of BT ratio, 3respectively, by parameter and entity (see Figure 3 for corresponding box plots) Supplemental Table 7. Monte Carlo cross-validation results Supplemental Table 8. Cases rejected prior to study inclusion Supplemental Table 9: Markers representing predominantly background signal (BS) by entity, Peer reviewed

1 figure., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed

1 figure., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed

1 figure., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed

1 figure., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed

1 figure., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed

Table 1. ‘FC50xFD54’ VOCs content. Relative average and standard deviation content of 58 VOCs detected in ‘FC50’, ‘FCD54’, and their F1 progeny in 6 harvests. VOCs name, KVCs mark, its abbreviation and compound family. Average, standard deviation and range of population and correlation between harvests in each VOC., Cultivated strawberry, Fragaria  ×  ananassa, has a complex aroma due to the presence of more than 350 volatile organic compounds (VOCs). However, a mixture of only 19 compounds, called Key Volatile Compounds (KVC), can impart the main strawberry aroma. The octoploid nature of the cultivated strawberry species (2n = 8x = 56) adds complexity to the heritance of the accumulation of the volatiles responsible for aroma. An F1 population cross between two breeding parental lines, FC50 and FD54, was phenotyped for aroma by SPME GCMS during six harvests. A total of 58 compounds were identified: 33 esters, nine terpenes, seven aldehydes, four lactones, two furans, one acid, one alkane and one alcohol, of which 16 were KVCs. A total of 179 QTLs were found, and 85 of these were detected in at least three harvests, of which 50 QTLs were considered major (LOD > 4.0) and detected in five or six analyzed harvests. Several clusters of ester QTLs associated with fruity aroma were discovered, such as QTLs for esters that share hexanoate group that were mapped in LG4A (Hexanoate_4A), those that share acetate and octyl groups in LG6A (Acetate_6A and Octyl_6A) or those with the same methyl group in LG7B (Methyl_7B). Different terpene QTLs associated with floral aroma appear grouped in a cluster in LG3C (Terpene_3C). Some of these clusters of QTLs were validated in a second F2 population, a cross of “Camarosa” and “Dover,” that was also phenotyped for three years. Selected SNPs from floral and fruity aroma QTLs were tested in a third population, which will most likely be useful for marker-assisted breeding (MAB)., Peer reviewed