BUSCADOR RECOLECTA

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/cancers14010192/s1, Figure S1: Bioinformatic analysis of 353 human HCC samples based on data generated by the Cancer Genome Atlas (TCGA) Research Network, Figure S2: Representative IHC-stained sections demonstrating nuclear expression of c-MYC. (A) c-MYC nuclear expression inside tumor nodules in patients with MAFLD-related HCC. Left: the DAB-stained images. Right: Hematoxylin stained images. (B) c-MYC nuclear expression in MAFLD patients with advanced liver fibrosis. Left: the DAB-stained images. Right: Hematoxylin stained images, Figure S3: MS in alb-myctg at baseline. WT (n = 9) and alb-myctg (n = 8) mice at the age of 36 weeks. (A) Food caloric intake was stable in both groups through the duration of study. (B) Representative MRI scan images of WT and alb-myctg mice showing body fat distribution in 26 weeks old animals. The quantitative data represent fat/body ratio (%) means. (C) Representative F4/80 IF staining of WAT. Positive cells are stained in green and marked with red arrows. Nuclei are stained in blue using DAPI as counterstain. Scale bar is 100 µm (n = 4), Figure S4: Glucose intolerance. (A) The histogram represents the incremental area under the respective glucose tolerance curve (n = 3). (B) Insulin tolerance test (ITT), and the histogram represents the incremental area under the curve (n = 3). Data are expressed as the mean ± SD, * = p < 0.05; ** = p < 0.01, alb-myctg mice vs. WT controls, Figure S5: Hepatosomatic ratio at 36 weeks. WT (n = 9) and alb-myctg (n = 8) mice at the age of 36 weeks. (A) Liver weight. (B) Liver to body weight ratio, Figure S6: Hepatic phenotype in WT and alb-myctg mice at 36 weeks. (A) qPCR analysis of hepatic mRNA expression expression of Scd1. (B) Histograms showing densitometric analysis of western blots for c-MYC, CYP2E1 and CHOP. (C) qPCR analysis of hepatic mRNA expression expression of Ucp2 (n = 4). (D) Representative TEM images of hepatic endoplasmic reticulum. Representative Transmission Electro-Microscopy (TEM) pictures of WT and alb-myctg mice. Asterisks indicate fat droplets in hepatocytes. Arrows mark endoplasmic reticulum (ER). Scale= 10 µm and 1 µm, Figure S7: Spontaneous proliferation and mild inflammation in alb-myctg mice at 36 weeks. (A) Histograms showing densitometric analysis of western blots for CC3 and PCNA. (B) Ki-67 immunofluorescence of crytosections showing increased cell proliferation (green, as indicated by arrow) of hepatocytes in alb-myctg mice liver and the quantification of % Ki-67 positive cells (n = 8). (C-E) Quantification of % CD45, F4/80- positive cells and SR area respectively using ImageJ software (n = 8). Data are expressed as the mean ± SD, * =p < 0.05; ** = p < 0.01, *** = p < 0.001, alb-myctg mice vs. WT controls, Figure S8: Increased glutamine catabolism in alb-myctg mice at 36 weeks. (n = 5). (A) IHC staining of Glutamine synthetase (GS). (B) Protein expression of Glutaminase 1 (GLS1) in the livers of alb-myctg and WT mice, Figure S9: Activation of signaling pathways relevant to human MAFLD in the liver of mice. (A) Dysregulated genes involved in different pathways. (B) Heat map demonstrating deregulated genes. Red and green colors indicate high and low gene expression, respectively. (C) Top canonical pathways. (D) Molecules involved in hepatotoxicity. All data are based on statistical significance, Figure S10: Spontaneous HCC development in alb-myctg mice. WT (n = 3) and alb-myctg (n = 5) mice at the age of 52 weeks. (A) Macroscopic images of the liver. Numbers represent tumor incidence (%) means (B) Representative liver sections stained with H&E, Figure S11: Phenotypical changes in WT and transgenic mice fed with WD for 24 weeks. (A) Growth curve demonstrating body weight gain, assessed at 7 days interval at each group (B) Relevant adipocyte area of eWAT (n = 6). (C) Representative F4/80 IF staining of WAT. Positive cells are stained in green and marked with red arrows. Nuclei are stained in blue using DAPI as counterstain. Scale bar is 100 µm (n = 4). (D) Quantification of Oil Red O positive area (n = 5). (E) Histological NAFLD activity score (n = 5). (F) Quantification of % CD45 positive cells (n = 6). (G) Quantification of % SR positive area (n = 4). (H) Quantification of % αSMA positive area (n = 3). (I) qPCR analysis of hepatic tissue of fibrosis gene αSma (n = 6). (J) Quantification of % KI-67 positive cells (n = 6). (K) Histograms showing densitometric analysis of western blot for CC3. Data are expressed as the mean ± SD, * = p < 0.05; ** = p < 0.01, *** = p < 0.001, alb-myctg mice vs. WT animals fed WD, Figure S12: MS and phenotypical changes in animals fed with metformin enriched diet. (A) Growth curve demonstrating body weight gain of mice, assessed at 7 days interval at each group, and (B) Relevant histogram represents the incremental area under the respective body weight curve. (C) Level of AST in serum (n = 6–8). (D) Quantification of % Ki-67 positive cells (n = 6). (E) qPCR analysis of hepatic mRNA expression expression of c-Myc. (F) Histograms showing densitometric analysis of western blots for SREBP1. Data are expressed as the mean ± SD, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, alb-myctg mice fed metformin enriched chow diet vs. alb-myctg mice fed chow diet; * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001, WT mice fed metformin enriched chow diet vs. alb-myctg mice fed chow diet, Figure S13: Uncropped blots for Figure 4C, Figure S14: Uncropped blots for Figure 4E, Figure S15: Uncropped blots for Figure 5I, Figure S16: Uncropped blots for Figure 7I, Table S1: Etiology of human liver samples used for c-MYC gene expression analysis, Table S2: Clinical features of human liver samples, Table S3: Kcal% of chow diet (CD) and western diet (WD), Table S4: Diets used for the development of MAFLD, Table S5: Antibodies used for IF staining, Table S6: Antibodies used for IHC and IF stainings, Table S7: Primers for qPCR, Table S8: List of antibodies used for WB., Peer reviewed

Table 1. Summary of PROM data over the different time points., Dysferlinopathy is a muscular dystrophy with a highly variable functional disease progression in which the relationship of function to some patient reported outcome measures (PROMs) has not been previously reported. This analysis aims to identify the suitability of PROMs and their association with motor performance.Two-hundred and four patients with dysferlinopathy were identified in the Jain Foundation's Clinical Outcome Study in Dysferlinopathy from 14 sites in 8 countries. All patients completed the following PROMs: Individualized Neuromuscular Quality of Life Questionnaire (INQoL), International Physical Activity Questionnaire (IPAQ), and activity limitations for patients with upper and/or lower limb impairments (ACTIVLIMs). In addition, nonambulant patients completed the Egen Klassifikation Scale (EK). Assessments were conducted annually at baseline, years 1, 2, 3, and 4. Data were also collected on the North Star Assessment for Limb Girdle Type Muscular Dystrophies (NSAD) and Performance of Upper Limb (PUL) at these time points from year 2. Data were analyzed using descriptive statistics and Rasch analysis was conducted on ACTIVLIM, EK, INQoL. For associations, graphs (NSAD with ACTIVLIM, IPAQ and INQoL and EK with PUL) were generated from generalized estimating equations (GEE). The ACTIVLIM appeared robust psychometrically and was strongly associated with the NSAD total score (Pseudo R2 0.68). The INQoL performed less well and was poorly associated with the NSAD total score (Pseudo R2 0.18). EK scores were strongly associated with PUL (Pseudo R2 0.69). IPAQ was poorly associated with NSAD scores (Pseudo R2 0.09). This study showed that several of the chosen PROMs demonstrated change over time and a good association with functional outcomes. An alternative quality of life measure and method of collecting data on physical activity may need to be selected for assessing dysferlinopathy., Peer reviewed

Appendix B. Coinvestigators - The Jain COS Consortium., Dysferlinopathy is a muscular dystrophy with a highly variable functional disease progression in which the relationship of function to some patient reported outcome measures (PROMs) has not been previously reported. This analysis aims to identify the suitability of PROMs and their association with motor performance.Two-hundred and four patients with dysferlinopathy were identified in the Jain Foundation's Clinical Outcome Study in Dysferlinopathy from 14 sites in 8 countries. All patients completed the following PROMs: Individualized Neuromuscular Quality of Life Questionnaire (INQoL), International Physical Activity Questionnaire (IPAQ), and activity limitations for patients with upper and/or lower limb impairments (ACTIVLIMs). In addition, nonambulant patients completed the Egen Klassifikation Scale (EK). Assessments were conducted annually at baseline, years 1, 2, 3, and 4. Data were also collected on the North Star Assessment for Limb Girdle Type Muscular Dystrophies (NSAD) and Performance of Upper Limb (PUL) at these time points from year 2. Data were analyzed using descriptive statistics and Rasch analysis was conducted on ACTIVLIM, EK, INQoL. For associations, graphs (NSAD with ACTIVLIM, IPAQ and INQoL and EK with PUL) were generated from generalized estimating equations (GEE). The ACTIVLIM appeared robust psychometrically and was strongly associated with the NSAD total score (Pseudo R2 0.68). The INQoL performed less well and was poorly associated with the NSAD total score (Pseudo R2 0.18). EK scores were strongly associated with PUL (Pseudo R2 0.69). IPAQ was poorly associated with NSAD scores (Pseudo R2 0.09). This study showed that several of the chosen PROMs demonstrated change over time and a good association with functional outcomes. An alternative quality of life measure and method of collecting data on physical activity may need to be selected for assessing dysferlinopathy., Peer reviewed

The available data contains: sky images before, during, and after the flash, event file, corresponding ARF and RMF, light-curves. See README file for further information., Peer reviewed

Figure S1. Distribution of z-scores for individual items across patients grouped by major domains (i.e., each count represents the score of one item in one of the patients’ assessment) Values less than 0 indicate items where brain tumour patients showed lower performance than healthy controls. Table S1. Individual cognitive tests included in the OCS-Bridge assessment and the traditional neuropsychology battery. Table S2. Combined domains across neuropsychological testing and OCS-Bridge testing. Table S3. OCS-Bridge screening and neuropsychological assessments completed by each participant. Table S4. Average total number of cognitive deficits and standard deviations across patients for each domain and assessment. A positive T value indicates a larger number of deficits detected by traditional neuropsychology compared with OCS-BRIDGE., Peer reviewed

1 table. -- Real activity (measured at physiological conditions) and total activity (measured at optimal enzymatic conditions) of the major digestive enzymes in the stomach (pepsin) and pyloric caeca (trypsin, chymotrypsin, leucine aminopeptidase and lipase) of S. dumerili., The study of fish digestive biochemistry is essential to understand factors that affect the net efficiency of food transformation and growth, and therefore aquaculture profitability. The aim of the present study was to assess the activity and functional characteristics of key digestive enzymes in juveniles of greater amberjack (Seriola dumerili), as well as the possible modulation of their relative importance by water temperature. For that, a combination of biochemical assays and substrate-SDS-PAGE were used. Under physiological conditions pepsin activity was negligible. Chymotrypsin was the most active enzyme in the digestive tract of the greater amberjack, while lipase was the enzyme with lower activity, though both enzymes in addition to trypsin were responsive to water temperature as revealed by discriminant analysis. Seriola dumerili showed to have pH-sensitive and, except for chymotrypsin, thermally robust proteases. Inhibition assays showed the major importance of serine proteases and revealed inverse trypsin and chymotrypsin responses to environmental temperature, with higher trypsin contribution in 26°C-fish while higher chymotrypsin contribution in 18°C-fish. Zymograms revealed three isotrypsin and three isochymotrypsin enzymes, with no variation in the presence of particular isoforms among rearing temperatures. However, they confirmed the role of chymotrypsin activity in providing digestive plasticity, with one of the isoforms being more active at lower temperatures. Thus, results indicate that variation in the relative contribution of chymotrypsin isoenzymes to a particular environmental temperature occurs due to different physic-chemical features of isoforms as a source of functional flexibility. This study assessed for the first time the effects of rearing temperature on greater amberjack digestive enzymes, increasing the knowledge on its digestive biochemistry, and aiding in the improvement of management practices for this species industrialization., Peer reviewed

The extreme currents matrix E and the explicite xtreme currents of KNS-LES, KNSCI and KNSCI-LES models., Peer reviewed

1 figure. -- Effect of Ph on the stability of trypsin (TRY), chymotripsin (CHY), leucine aminopeptidase (LAP), lipase (LIP) and pepsin (PP) activities from pyloric caeca and stomach of S. Dumerili adapted to different rearing temperatures (18, 22 and 26º C)., The study of fish digestive biochemistry is essential to understand factors that affect the net efficiency of food transformation and growth, and therefore aquaculture profitability. The aim of the present study was to assess the activity and functional characteristics of key digestive enzymes in juveniles of greater amberjack (Seriola dumerili), as well as the possible modulation of their relative importance by water temperature. For that, a combination of biochemical assays and substrate-SDS-PAGE were used. Under physiological conditions pepsin activity was negligible. Chymotrypsin was the most active enzyme in the digestive tract of the greater amberjack, while lipase was the enzyme with lower activity, though both enzymes in addition to trypsin were responsive to water temperature as revealed by discriminant analysis. Seriola dumerili showed to have pH-sensitive and, except for chymotrypsin, thermally robust proteases. Inhibition assays showed the major importance of serine proteases and revealed inverse trypsin and chymotrypsin responses to environmental temperature, with higher trypsin contribution in 26°C-fish while higher chymotrypsin contribution in 18°C-fish. Zymograms revealed three isotrypsin and three isochymotrypsin enzymes, with no variation in the presence of particular isoforms among rearing temperatures. However, they confirmed the role of chymotrypsin activity in providing digestive plasticity, with one of the isoforms being more active at lower temperatures. Thus, results indicate that variation in the relative contribution of chymotrypsin isoenzymes to a particular environmental temperature occurs due to different physic-chemical features of isoforms as a source of functional flexibility. This study assessed for the first time the effects of rearing temperature on greater amberjack digestive enzymes, increasing the knowledge on its digestive biochemistry, and aiding in the improvement of management practices for this species industrialization., Peer reviewed

eAppendix. Additional acknowledgments eMethods. eFigure 1. Flowchart describing the number of individuals remaining at each filtering steps eFigure 2. V236E and R251G are associated with decreased AD risk across dataset in APOE-stratified sensitivity analyses eFigure 3. APOE ε3/ε3[V236E] individuals have a lower AD risk than APOE ε2/ε3 individuals and APOE ε3/ε4[R251G] have a risk equivalent to ε2/ε3 carriers despite carrying 1 ε4 allele, regardless of the EUR ancestry cutoff for admixed Europeans and Europeans eTable 1. Queried cohort overview to identify admixed and European ancestry individuals in the ADSP discovery and ADGC internal replication eTable 2. Overview of ADSP studies with whole-exome sequencing (WES) and/or whole-genome sequencing (WGS) available at NIAGADS DSS (NG00067) eTable 3. Demographic characteristics of the cohorts queried for discovery and internal replication samples eTable 4. Missense variants on the APOE canonical transcript reported in gnomADv.3.1 eTable 5. Demographic characteristics per cohort in ADSP discovery and ADGC internal replication after ancestry selection, quality control, and duplicates removal eTable 6. APOE missense variants rs769452-C (APOE[L28P]), rs199768005-A (APOE[V236E]), and rs267606661-G (APOE[R251G]) allelic breakdown by APOE main genotype eTable 7. V236E and R251G association in primary and secondary analyses, nonstratified and APOE stratified eTable 8. Nonstratified sensitivity analyses at various European ancestry cutoffs eTable 9. Sensitivity analysis, including all dementia in the CCHS and CGPS data set, slightly strengthens the V236E and R251G associations with decreased AD risk eTable 10. Sensitivity analysis, excluding the UK Biobank proxy-AD phenotype from the meta-analysis, results in slight worsening of the P values of the V236E and R251G associations with decreased AD risk eReferences. Supplemental Online Content: Nonauthor Collaborators., Data for this study were prepared, archived, and distributed by the National Institute on Aging Alzheimer’s Disease Data Storage Site (NIAGADS) at the University of Pennsylvania (U24-AG041689), funded by the National Institute on Aging., Peer reviewed

1 figure. -- Effect of Ph on the stability of trypsin (TRY), chymotripsin (CHY), leucine aminopeptidase (LAP), lipase (LIP) and pepsin (PP) activities from pyloric caeca and stomach of S. Dumerili adapted to different rearing temperatures (18, 22 and 26º C)., The study of fish digestive biochemistry is essential to understand factors that affect the net efficiency of food transformation and growth, and therefore aquaculture profitability. The aim of the present study was to assess the activity and functional characteristics of key digestive enzymes in juveniles of greater amberjack (Seriola dumerili), as well as the possible modulation of their relative importance by water temperature. For that, a combination of biochemical assays and substrate-SDS-PAGE were used. Under physiological conditions pepsin activity was negligible. Chymotrypsin was the most active enzyme in the digestive tract of the greater amberjack, while lipase was the enzyme with lower activity, though both enzymes in addition to trypsin were responsive to water temperature as revealed by discriminant analysis. Seriola dumerili showed to have pH-sensitive and, except for chymotrypsin, thermally robust proteases. Inhibition assays showed the major importance of serine proteases and revealed inverse trypsin and chymotrypsin responses to environmental temperature, with higher trypsin contribution in 26°C-fish while higher chymotrypsin contribution in 18°C-fish. Zymograms revealed three isotrypsin and three isochymotrypsin enzymes, with no variation in the presence of particular isoforms among rearing temperatures. However, they confirmed the role of chymotrypsin activity in providing digestive plasticity, with one of the isoforms being more active at lower temperatures. Thus, results indicate that variation in the relative contribution of chymotrypsin isoenzymes to a particular environmental temperature occurs due to different physic-chemical features of isoforms as a source of functional flexibility. This study assessed for the first time the effects of rearing temperature on greater amberjack digestive enzymes, increasing the knowledge on its digestive biochemistry, and aiding in the improvement of management practices for this species industrialization., Peer reviewed