A Descriptive Study of Spanish and Ecuadorian Commercial Infant Cereals: Are They in Line with Current Recommendations?

Cereals are an important source of nutrients, especially used in complementary feeding. The objective of this study is to review the nutritional composition of cereal-based foods for infants from 4 months and toddlers that are offered in Spain and Ecuador, countries selected because of the opportunity to work in them, and due to their socio-economic differences (industrialized and developing countries, respectively). The number of these products was 105 cereals in Spain and 22 in Ecuador. The products were classified as gluten-free cereals, five cereals, eight cereals, multigrain cereals, and cookies. A 25 g serving was used to determine the percentage in which the samples analyzed can cover the Reference Nutrient Intake (RNI) for micronutrients in infants from 7 months and toddlers according to the European Food Safety Authority (EFSA). Nutritional information per 100 g of dry product was collected according to medium, minimum, and maximum units, and nutrient density was calculated. The age range in which these products are recommended is different in both countries. The nutritional composition presents some differences; Spanish cereals show a lower content of sodium, added sugars, hydrolyzed cereals, and maltodextrin than Ecuadorian cereals. Commercialized cereals could contribute to satisfying the nutritional needs of infants and toddlers; however, they can also be a source of non-recommended components.

Analysis of Susceptibility or Resistance to Antimicrobial Agents.

Here were described the main three methods being used for analysis of antibiotic susceptibility or resistance of Streptococcus suis clinical isolates to antimicrobial agents: the Kirby-Bauer disk diffusion, the epsilometer test (E test), and the broth microdilution test. In each case, procedures, results, and interpretation are described, as well as their advantages or limitations when proceeds.

The Biodistribution of the Spike Protein after Ad26.COV2.S Vaccination Is Unlikely to Play a Role in Vaccine-Induced Immune Thrombotic Thrombocytopenia.

Ad26.COV2.S vaccination can lead to vaccine-induced immune thrombotic thrombocytopenia (VITT), a rare but severe adverse effect, characterized by thrombocytopenia and thrombosis. The mechanism of VITT induction is unclear and likely multifactorial, potentially including the activation of platelets and endothelial cells mediated by the vaccine-encoded spike protein (S protein). Here, we investigated the biodistribution of the S protein after Ad26.COV2.S dosing in three animal models and in human serum samples. The S protein was transiently present in draining lymph nodes of rabbits after Ad26.COV2.S dosing. The S protein was detected in the serum in all species from 1 day to 21 days after vaccination with Ad26.COV2.S, but it was not detected in platelets, the endothelium lining the blood vessels, or other organs. The S protein S1 and S2 subunits were detected at different ratios and magnitudes after Ad26.COV2.S or COVID-19 mRNA vaccine immunization. However, the S1/S2 ratio did not depend on the Ad26 platform, but on mutation of the furin cleavage site, suggesting that the S1/S2 ratio is not VITT related. Overall, our data suggest that the S-protein biodistribution and kinetics after Ad26.COV2.S dosing are likely not main contributors to the development of VITT, but other S-protein-specific parameters require further investigation.

Peak transgene expression after intramuscular immunization of mice with adenovirus 26-based vector vaccines correlates with transgene-specific adaptive immune responses.

Non-replicating adenovirus-based vectors have been broadly used for the development of prophylactic vaccines in humans and are licensed for COVID-19 and Ebola virus disease prevention. Adenovirus-based vectored vaccines encode for one or more disease specific transgenes with the aim to induce protective immunity against the target disease. The magnitude and duration of transgene expression of adenovirus 5- based vectors (human type C) in the host are key factors influencing antigen presentation and adaptive immune responses. Here we characterize the magnitude, duration, and organ biodistribution of transgene expression after single intramuscular administration of adenovirus 26-based vector vaccines in mice and evaluate the differences with adenovirus 5-based vector vaccine to understand if this is universally applicable across serotypes. We demonstrate a correlation between peak transgene expression early after adenovirus 26-based vaccination and transgene-specific cellular and humoral immune responses for a model antigen and SARS-CoV-2 spike protein, independent of innate immune activation. Notably, the memory immune response was similar in mice immunized with adenovirus 26-based vaccine and adenovirus 5-based vaccine, despite the latter inducing a higher peak of transgene expression early after immunization and a longer duration of transgene expression. Together these results provide further insights into the mode of action of adenovirus 26-based vector vaccines.

Streptococcus suis Research Update: Serotype Prevalence and Antimicrobial Resistance Distribution in Swine Isolates Recovered in Spain from 2020 to 2022.

This study aimed to update the Streptococcus suis serotype distribution in Spain by analysing 302 clinical isolates recovered from diseased pigs between 2020 and 2022. The main objectives were to identify prevalent serotypes, differentiate specific serotypes 1, 14, 2, and 1/2, investigate specific genotypic and phenotypic antimicrobial resistance features, and explore associations between resistance genes and phenotypic resistances. Serotypes 9 (21.2%), 1 (16.2%), 2 (15.6%), 3 (6%), and 7 (5.6%) were the most prevalent, whereas serotypes 14 and 1/2 corresponded with 4.3% and 0.7% of all isolates. Antimicrobial resistance genes, including tet(O), erm(B), lnu(B), lsa(E), tet(M), and mef(A/E), were analysed, which were present in 85.8%, 65.2%, 7%, 7%, 6.3%, and 1% of the samples, respectively. Susceptibility testing for 18 antimicrobials revealed high resistance levels, particularly for clindamycin (88.4%), chlortetracycline (89.4%), and sulfadimethoxine (94.4%). Notably, seven significant associations (p < 0.0001) were detected, correlating specific antimicrobial resistance genes to the observed phenotypic resistance. These findings contribute to understanding the S. suis serotype distribution and its antibiotic resistance profiles in Spain, offering valuable insights for veterinary and public health efforts in managing S. suis-associated infections.

Brazilian Clinical Strains of Actinobacillus pleuropneumoniae and Pasteurella multocida: Capsular Diversity, Antimicrobial Susceptibility (In Vitro) and Proof of Concept for Prevention of Natural Colonization by Multi-Doses Protocol of Tildipirosin.

One hundred Actinobacillus pleuropneumoniae (App) and sixty Pasteurella multocida subsp. multocida serogroup A (PmA) isolates were recovered from porcine pneumonic lungs collected from eight central or southern states of Brazil between 2014 and 2018 (App) or between 2017 and 2021 (PmA). A. pleuropneumoniae clinical isolates were typed by multiplex PCR and the most prevalent serovars were 8, 7 and 5 (43, 25% and 18%, respectively). In addition, three virulence genes were assessed in P. multocida isolates, all being positive to capA (PmA) and kmt1 genes, all negative to capD and toxA, and most of them (85%) negative to pfhA gene. The susceptibility of both pathogens to tildipirosin was investigated using a broth microdilution assay. The percentage of isolates susceptible to tildipirosin was 95% for App and 73.3% for PmA. The MIC50 values were 0.25 and 1 µg/mL and the MIC90 values were 4 and >64 µg/mL for App and PmA, respectively. Finally, a multiple-dose protocol of tildipirosin was tested in suckling piglets on a farm endemic for both pathogens. Tildipirosin was able to prevent the natural colonization of the tonsils by App and PmA and significantly (p < 0.0001) reduced the burden of Glaesserella parasuis in this tissue. In summary, our results demonstrate that: (i) tildipirosin can be included in the list of antibiotics to control outbreaks of lung disease caused by App regardless of the capsular type, and (ii) in the case of clinical strains of App and PmA that are sensitive to tildipirosin based on susceptibility testing, the use of this antibiotic in eradication programs for A. pleuropneumoniae and P. multocida can be strongly recommended.

Intravenous Administration of Ad26.COV2.S Does Not Induce Thrombocytopenia or Thrombotic Events or Affect SARS-CoV-2 Spike Protein Bioavailability in Blood Compared with Intramuscular Vaccination in Rabbits.

Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a very rare but serious adverse reaction that can occur after Ad26.COV2.S vaccination in humans, leading to thrombosis at unusual anatomic sites. One hypothesis is that accidental intravenous (IV) administration of Ad26.COV2.S or drainage of the vaccine from the muscle into the circulatory system may result in interaction of the vaccine with blood factors associated with platelet activation, leading to VITT. Here, we demonstrate that, similar to intramuscular (IM) administration of Ad26.COV2.S in rabbits, IV dosing was well tolerated, with no significant differences between dosing routes for the assessed hematologic, coagulation time, innate immune, or clinical chemistry parameters and no histopathologic indication of thrombotic events. For both routes, all other non-adverse findings observed were consistent with a normal vaccine response and comparable to those observed for unrelated or other Ad26-based control vaccines. However, Ad26.COV2.S induced significantly higher levels of C-reactive protein on day 1 after IM vaccination compared with an Ad26-based control vaccine encoding a different transgene, suggesting an inflammatory effect of the vaccine-encoded spike protein. Although based on a limited number of animals, these data indicate that an accidental IV injection of Ad26.COV2.S may not represent an increased risk for VITT.

Machine learning identifies experimental brain metastasis subtypes based on their influence on neural circuits.

A high percentage of patients with brain metastases frequently develop neurocognitive symptoms; however, understanding how brain metastasis co-opts the function of neuronal circuits beyond a tumor mass effect remains unknown. We report a comprehensive multidimensional modeling of brain functional analyses in the context of brain metastasis. By testing different preclinical models of brain metastasis from various primary sources and oncogenic profiles, we dissociated the heterogeneous impact on local field potential oscillatory activity from cortical and hippocampal areas that we detected from the homogeneous inter-model tumor size or glial response. In contrast, we report a potential underlying molecular program responsible for impairing neuronal crosstalk by scoring the transcriptomic and mutational profiles in a model-specific manner. Additionally, measurement of various brain activity readouts matched with machine learning strategies confirmed model-specific alterations that could help predict the presence and subtype of metastasis.

Objective measured physical activity and metabolic syndrome score in children and adolescents: The UP&DOWN longitudinal study.

INTRODUCTION: We aimed to analyze the cross-sectional and longitudinal association of physical activity (PA) levels and PA patterns with metabolic syndrome score (MetS) in children and adolescents. METHODS: A total of 175 children (82 females) and 188 adolescents (95 females) were included. Objective PA levels and patterns were determined by accelerometry. MetS was computed from waist circumference, systolic blood pressure, triglycerides, high-density lipoprotein cholesterol, and glucose levels. Different linear regression models were implemented to examine the associations of PA with MetS. RESULTS: Vigorous PA, moderate-vigorous PA, number of bouts per day in 10 min (N10), and total time in bouts per day in 10 min (T10) were negatively associated with MetS in male children and adolescents at cross-sectional level (ß ranging from -0.005 to -0.164, all p < 0.05). Total time in bouts per day in 20 min in male children, and vigorous PA and N10 in female children were longitudinally and negatively associated with MetS (ß ranging from -0.011 to -0.247, all p < 0.05). CONCLUSIONS: Associations of PA and MetS were observed at cross-sectional level in males and longitudinally in female children. The associations in PA patterns were found when patterns were grouped into bouts of 10 min. Therefore, for future studies of PA with health markers in the pediatric population, it would be advisable to choose bouts of shorter duration.

Longitudinal reallocations of time between 24-h movement behaviours and their associations with inflammation in children and adolescents: the UP&DOWN study.

BACKGROUND: While there is evidence that physical activity, sedentary behaviour (SB) and sleep may all be associated with modified levels of inflammatory markers in adolescents and children, associations with one movement behaviour have not always been adjusted for other movement behaviours, and few studies have considered all movement behaviours in the 24-hour day as an exposure. PURPOSE: The aim of the study was to explore how longitudinal reallocations of time between moderate-to-vigorous physical activity (MVPA), light physical activity (LPA), SB and sleep are associated with changes in inflammatory markers in children and adolescents. METHODS: A total of 296 children/adolescents participated in a prospective cohort study with a 3-year follow-up. MVPA, LPA and SB were assessed by accelerometers. Sleep duration was assessed using the Health Behavior in School-aged Children questionnaire. Longitudinal compositional regression models were used to explore how reallocations of time between movement behaviours are associated with changes in inflammatory markers. RESULTS: Reallocations of time from SB to sleep were associated with increases in C3 levels (difference for 60 min/d reallocation [d60] = 5.29 mg/dl; 95% confidence interval [CI] = 0.28, 10.29) and TNF-α (d60 = 1.81 mg/dl; 95% CI = 0.79, 15.41) levels. Reallocations from LPA to sleep were also associated with increases in C3 levels (d60 = 8.10 mg/dl; 95% CI = 0.79, 15.41). Reallocations from LPA to any of the remaining time-use components were associated with increases in C4 levels (d60 ranging from 2.54 to 3.63 mg/dl; p < 0.05), while any reallocation of time away from MVPA was associated with unfavourable changes in leptin (d60 ranging from 3088.44 to 3448.07 pg/ml; p < 0.05). CONCLUSIONS: Reallocations of time between 24-h movement behaviours are prospectively associated with some inflammatory markers. Reallocating time away from LPA appears to be most consistently unfavourably associated with inflammatory markers. Given that higher levels of inflammation during childhood and adolescence are associated with an increased risk of chronic diseases in adulthood, children and adolescents should be encouraged to maintain or increase the level of LPA to preserve a healthy immune system.

Programa de Mentoría: acompañando tus primeros pasos hacia la excelencia científica / Mentoring Programme: Guiding your first steps towards scientific excellence

El primer Programa de Mentoría de SEMICYUC tiene como objetivo apoyar la carrera investigadora de los miembros más jóvenes de la Sociedad. Como beneficios añadidos está la adquisición de nuevas capacidades de investigación y/o clínicas, incrementar la capacidad de reflexión y fomentar el desarrollo de la próxima generación de líderes en la investigación. Este proyecto no sería posible sin el equipo excepcional de mentores o expertos investigadores dispuestos a emprender el viaje con los jóvenes aprendices. El presente artículo expone las bases de dicho programa, además de proponer futuros cambios en haz de una mejora continua (AU)

SEMICYUC's first Mentoring Programme aims to support the research careers of the Society's youngest members. Added benefits include acquiring new research and/or clinical skills, increasing the ability of critical thought, and fostering the development of the next generation of research leaders. This project would not be possible without the exceptional team of mentors or research experts willing to embark on the journey with the young trainees. This article sets out the foundations of such a programme and proposes future changes for continuous improvement (AU)

Phylogenetic study and comparison of different TbpB obtained from Glaesserella parasuis present in Spanish clinical isolates.

Glaesserella parasuis (Gp) is the etiological agent of Glässer's disease (GD), which causes important economic losses for the pig intensive production worldwide. This organism uses a smart protein-based receptor to acquire specifically iron from the porcine transferrin. This surface receptor consists of transferrin-binding protein A (TbpA) and transferrin-binding protein B (TbpB). TbpB has been considered the most promising antigen to formulate a based-protein vaccine with broad-spectrum of protection against GD. The purpose of our study was to determine the capsular diversity of Gp clinical isolates collected in different Spanish regions between 2018 and 2021. A total of 68 Gp isolates were recovered from porcine respiratory or systemic samples. A species-specific PCR based on tbpA gene, followed by multiplex PCR for typing Gp isolates were performed. Serovars 5, 10, 2, 4 and 1 were the most prevalent and involved almost 84% of isolates. TbpB amino acid sequences from 59 of these isolates were analyzed, and a total of ten clades could be established. All of them showed a wide diversity with respect to capsular type, anatomical isolation site and geographical origin, with minor exceptions. Regardless of the serovars, the in silico analysis of TbpB sequences revealed that a vaccine based on a TbpB recombinant protein could potentially prevent Glässer's disease outbreaks in Spain.

Single layer centrifugation (SLC) for bacterial removal with Porcicoll positively modifies chromatin structure in boar spermatozoa.

The storage of boar semen samples at 17 °C for artificial insemination (AI) doses enables the proliferation of the bacteria, making antibiotics necessary. This can contribute to the development of antimicrobial resistance (AMR). This study tested bacterial presence and sperm chromatin structure after using a low-density colloid (Porcicoll) as an antibiotic alternative to eliminate bacteria. Ejaculates (8 boars, 3 ejaculates each) were split as control and low-density colloid centrifugation (single layer centrifugation, SLC, 20%, and 30% Porcicoll) into 500 ml tubes. Analyses were carried out at days 0, 3, and 7 (17 °C) for microbial presence and sperm chromatin structure analysis: %DFI (DNA fragmentation) and %HDS (chromatin immaturity), monobromobimane (mBBr; free thiols and disulfide bridges), and chromomycin A3 (CMA3; chromatin compaction). Besides comparing bacterial presence (7 species identified) and chromatin variables between treatments, the associations between these sets of variables were described by canonical correlation analysis (CCA). Results showed a significant decrease of some bacteria or a complete removal after SLC (especially for P30). SLC also caused a decrease of %HDS and an increase of disulfide bridges and low and medium mBBr populations, suggesting the removal of immature sperm (poor chromatin compaction). CCA showed an association pattern compatible with the degradation of sperm chromatin parameters with bacterial contamination, especially Enterobacteria, P. aeuriginosa, and K. variicola. In conclusion, bacterial contamination affects sperm chromatin beyond DNA fragmentation; SLC with low-density colloid not only removes bacteria from boar semen, but also chromatin structure is enhanced after selection.

Mentoring Programme: Guiding your first steps towards scientific excellence.

SEMICYUC's first Mentoring Programme aims to support the research careers of the Society's youngest members. Added benefits include acquiring new research and/or clinical skills, increasing the ability of critical thought, and fostering the development of the next generation of research leaders. This project would not be possible without the exceptional team of mentors or research experts willing to embark on the journey with the young trainees. This article sets out the foundations of such a programme and proposes future changes for continuous improvement.

New indices in predicting cardiometabolic risk and its relation to endothelial dysfunction in adolescents: The HELENA study.

BACKGROUND AND AIMS: Blood pressure (BP) changes and insulin resistance (IR) are important cardiometabolic risk (CMR) factors; their early identification can contribute to the reduction of cardiovascular events in adulthood. This necessitates the search for more accessible and easily applied indicators for their prediction. Therefore, this study aimed to evaluate the predictive power of the indices, TyG, TG/HDL-c, height-corrected lipid accumulation product (HLAP), and visceral adiposity index (VAI), in identifying the CMR obtained by high BP and IR and to verify their relationship with biomarkers of endothelial dysfunction (ED) in European adolescents. METHODS AND RESULTS: The anthropometric data and blood biomarkers of 744 adolescents (343 boys and 401 girls) from the Healthy Lifestyle in Europe by Nutrition in Adolescence Cross-Sectional Study (HELENA-CSS), with a mean age of 14.67 (SD 1.15) years, were assessed. The adolescents were then classified according to the presence or absence of high BP and IR. The cut-off points of the indices evaluated for the identification of CMR were determined. The relationship between CMR diagnosed using these indices and ED biomarkers was tested. The HLAP and TG/HDL-c were fair predictors of CMR obtained by IR in male adolescents. These indices showed association with hsCRP in sVCAM-1 in boys, but it lost significance after adjusting for age and body mass index. CONCLUSION: TG/HDL-c and HLAP indices showed a fair performance in predicting CMR, obtained by IR, in male adolescents. ED showed no association with the CMR identified by the indices.

Influence of Executive Function Training on BMI, Food Choice, and Cognition in Children with Obesity: Results from the TOuCH Study.

BACKGROUND: Children with obesity have a higher risk of future health and psychological problems. Executive functions (EFs) play a key role in successful dietetic and exercise planning; therefore, new treatments aimed at improving EFs may optimize outcomes. OBJECTIVES: This study evaluates the impact of EF training on body mass index (BMI), food choice, and cognition in children with obesity. We also examine their real-life executive functioning, emotional state, and quality of life. METHODS: Randomized controlled double-blind trial. Forty-six children with obesity were randomly allocated into an executive functions training or a control task training group and attended 30-45 min of daily training (5/week over 6 weeks), with both groups receiving counseling on diet and wearing an activity/sleep tracker. Participants were evaluated at baseline and after treatment. RESULTS: BMI decreased over time in the whole sample, although there were no differences between groups at post-training in BMI, food choice, and cognition. Both groups showed significant improvements in attention, speed, cognitive flexibility, and inhibitory control. Additionally, there were some benefits in real-life executive functioning and self-esteem. Over the 6 weeks, participants showed worse food choices in both groups. CONCLUSIONS: EFs training showed a lack of significant effects. The executive function enhancement alone did not explain these changes, as there were no significant differences between the experimental groups. It might be that the control task training could also produce some benefits, and multi-component interventions might be useful for weight loss.

Association between Moderate Alcohol Consumption and Subjective Quality of Life in Spanish Young Adults.

BACKGROUND: For the last 25 years, the debate on the benefit-risk balance of moderate alcohol consumption has been ongoing. This study explored the relationships between the pattern of alcohol consumption and subjective quality of life in healthy adults. MATERIAL AND METHODS: Participants were 247 healthy adults aged 25-45 years, with a moderate alcohol consumption, classified in three groups of alcohol intake: None (N = 37; <0.7 g/day), Low (N = 87; 0.7-<5 g/day) and Medium (N = 123; 5-16 g/day in women and 5-28 g/day in men). Information was collected with questionnaires including: alcoholic beverage frequency and pattern, quality of life (SF-36v2), anxiety, depression, health condition, adherence to the Mediterranean diet, physical activity and sleep time. RESULTS: Mean (SD) age of participants was 35.9 (6.3). In the Medium group, the mean alcohol intake was 10.98 ± 4.84 g/day on weekly bases and 24.7 ± 15.2 g/day on weekend days. Beer was the main contributor to total daily alcohol consumption. The percentage of subjects in the Medium group that showed a weekend average intake higher than moderate was 58.5% in exclusively weekend consumers and 48.2% in those who are not exclusive-weekend consumers (p = 0.278). Regarding markers of quality of life and mental health, the results did not show any significant association with alcohol consumption. In conclusion, in subjects that show weekly bases moderate intake of alcohol, weekend-day consumption levels can be high; however, no association of an overall moderate alcohol consumption pattern with quality of life was found.

Cell entry and innate sensing shape adaptive immune responses to adenovirus-based vaccines.

Nonreplicating adenovirus-based vectors have been successfully implemented as prophylactic vaccines against infectious viral diseases and induce protective cellular and humoral responses. Differences in the mechanisms of cellular entry or endosomal escape of these vectors contribute to differences in innate immune sensing between adenovirus species. Innate immune responses to adenovirus-based vaccines, such as interferon signaling, have been reported to affect the development of adaptive responses in preclinical studies, although limited data are available in humans. Understanding the mechanisms of these early events is critical for the development of vaccines that elicit effective and durable adaptive immune responses while maintaining an acceptable reactogenicity profile.

Food portion sizes and their relationship with energy, and nutrient intakes in adolescents: The HELENA study.

OBJECTIVES: This study aimed to investigate the associations between portion sizes (PSs) from different food groups and energy, as well as nutrient intakes in European adolescents. METHODS: A sample of 1631 adolescents (54.2 % girls) were included from the Healthy Lifestyle in Europe by Nutrition in Adolescence Cross-Sectional (HELENA) study. Mean food PS was calculated by dividing the total intake of the items by the number of eating occasions of these consumed items. To determine the key items for analysis, foods were ranked by frequency of consumption. A one-way between-groups analysis of covariance was used to test for significant differences in means across tertiles. A multivariable linear regression analysis was carried out, adjusting for age, sex, maternal education, body mass index, and using country as a level. RESULTS: Energy intake increased with elevated intakes of energy-dense foods. Large portions of rice and other grains, starch roots and potatoes, and meat substitutes, nuts, and pulses were associated with increased carbohydrate and fiber intake. Larger portions of cheese and butter and animal fat were significantly associated with a higher fat intake. Lower intakes of some vitamins and micronutrients were noticed with consumption of larger portions of high energy-dense foods, such as desserts and pudding, margarine and vegetable oil, and butter and animal fat. CONCLUSIONS: Large food PSs may be associated with positive energy, as well as macro- and micronutrient intake. Moreover, the findings from this study may help the future development of dietary guidance in general and specific to PSs, and support targeted strategies to address intakes of certain nutrients in European adolescents.

Fitness, body composition, and metabolic risk scores in children and adolescents: the UP&DOWN study.

We aimed to analyse the longitudinal association between physical fitness (PF) and body composition (BC) with a metabolic risk score (Met4) in children and adolescents and to elucidate whether the association between PF and Met4 differs when using relativized or absolute fitness variables. A total of 188 children (86 females) and 195 adolescents (97 females) were included. Cardiorespiratory fitness (CRF) was determined by the 20-m shuttle run test, and muscular fitness (MF) was determined by hand grip and standing long jump tests. Height and weight were measured, and the body mass index (Kg/m2) was calculated. Triceps and subscapular skinfolds were assessed to compute body fat percentage. Met4 was computed from systolic blood pressure, triglycerides, high-density lipoprotein cholesterol, and glucose levels. Relative CRF was longitudinally and negatively associated with Met4 in female children (ß = -0.031, p = 0.025), while absolute CRF was positively associated with Met4 in male children and adolescents (ß = 0.000, p < 0.05). Relative upper and lower-body MF were longitudinally and negatively associated with Met4 in female adolescents (ß = -1.347, ß = -0.005, p < 0.05), while absolute lower-body MF was positively associated with Met4 in male children (ß = 0.000, p = 0.019). BC was longitudinally and positively associated with Met4 in male children (ß-ranging from 0.011 to 0.055, all p < 0.05) and male adolescents (ß-ranging from 0.011 to 0.046, all p < 0.05).  Conclusion: BC is more strongly associated with Met4 than PF in children and adolescents. An optimal body weight status should be considered the main objective of health-promoting programs at childhood and adolescence. Furthermore, the way of expressing the fitness variables determines the direction of the association with Met4. What is Known: • Physical fitness is an important health indicator in children and adolescents, with great amount of previous evidence supporting the preventive role of maintaining optimal levels of both cardiorespiratory and muscular fitness for future cardiometabolic issues. What is New: • The way of reporting physical fitness variables can affect the associations between physical fitness features and cardiometabolic outcomes. Since body composition variables have a great impact on both physical fitness and cardiometabolic health, relativizing physical fitness performance by body composition could lead to erroneous conclusions.