Current infection biomarkers are highly limited since they have low capability to predict infection in the presence of confounding processes such as in non-infectious inflammatory processes, low capability to predict disease outcomes and have limited applications to guide and evaluate therapeutic regimes. Therefore, it is critical to discover and develop new and effective clinical infection biomarkers, especially applicable in patients at risk of developing severe illness and critically ill patients. Ideal biomarkers would effectively help physicians with better patient management, leading to a decrease of severe outcomes, personalize therapies, minimize antibiotics overuse and hospitalization time, and significantly improve patient survival. Metabolomics, by providing a direct insight into the functional metabolic outcome of an organism, presents a highly appealing strategy to discover these biomarkers. The present work reviews the desired main characteristics of infection biomarkers, the main metabolomics strategies to discover these biomarkers and the next steps for developing the area towards effective clinical biomarkers.

Biofluid metabolomics is a very appealing tool to increase the knowledge associated with pathophysiological mechanisms leading to better and new therapies and biomarkers for disease diagnosis and prognosis. However, due to the complex process of metabolome analysis, including the metabolome isolation method and the platform used to analyze it, there are diverse factors that affect metabolomics output. In the present work, the impact of two protocols to extract the serum metabolome, one using methanol and another using a mixture of methanol, acetonitrile, and water, was evaluated. The metabolome was analyzed by ultraperformance liquid chromatography associated with tandem mass spectrometry (UPLC-MS/MS), based on reverse-phase and hydrophobic chromatographic separations, and Fourier transform infrared (FTIR) spectroscopy. The two extraction protocols of the metabolome were compared over the analytical platforms (UPLC-MS/MS and FTIR spectroscopy) concerning the number of features, the type of features, common features, and the reproducibility of extraction replicas and analytical replicas. The ability of the extraction protocols to predict the survivability of critically ill patients hospitalized at an intensive care unit was also evaluated. The FTIR spectroscopy platform was compared to the UPLC-MS/MS platform and, despite not identifying metabolites and consequently not contributing as much as UPLC-MS/MS in terms of information concerning metabolic information, it enabled the comparison of the two extraction protocols as well as the development of very good predictive models of patient's survivability, such as the UPLC-MS/MS platform. Furthermore, FTIR spectroscopy is based on much simpler procedures and is rapid, economic, and applicable in the high-throughput mode, i.e., enabling the simultaneous analysis of hundreds of samples in the microliter range in a couple of hours. Therefore, FTIR spectroscopy represents a very interesting complementary technique not only to optimize processes as the metabolome isolation but also for obtaining biomarkers such as those for disease prognosis.

Background and Objectives: Given the wide spectrum of clinical and laboratory manifestations of the coronavirus disease 2019 (COVID-19), it is imperative to identify potential contributing factors to patients’ outcomes. However, a limited number of studies have assessed how the different waves affected the progression of the disease, more so in Portugal. Therefore, our main purpose was to study the clinical and laboratory patterns of COVID-19 in an unvaccinated population admitted to the intensive care unit, identifying characteristics associated with death, in each of the first three waves of the pandemic. Materials and Methods: This study included 337 COVID-19 patients admitted to the intensive care unit of a single-center hospital in Lisbon, Portugal, between March 2020 and March 2021. Comparisons were made between three COVID-19 waves, in the second (n = 325) and seventh (n = 216) days after admission, and between discharged and deceased patients. Results: Deceased patients were considerably older (p = 0.021) and needed greater ventilatory assistance (p = 0.023), especially in the first wave. Differences between discharged and deceased patients’ biomarkers were minimal in the first wave, on both analyzed days. In the second wave significant differences emerged in troponins, lactate dehydrogenase, procalcitonin, C-reactive protein, and white blood cell subpopulations, as well as platelet-to-lymphocyte and neutrophil-to-lymphocyte ratios (all p < 0.05). Furthermore, in the third wave, platelets and D-dimers were also significantly different between patients’ groups (all p < 0.05). From the second to the seventh days, troponins and lactate dehydrogenase showed significant decreases, mainly for discharged patients, while platelet counts increased (all p < 0.01). Lymphocytes significantly increased in discharged patients (all p < 0.05), while white blood cells rose in the second (all p < 0.001) and third (all p < 0.05) waves among deceased patients. Conclusions: This study yields insights into COVID-19 patients’ characteristics and mortality-associated biomarkers during Portugal’s first three COVID-19 waves, highlighting the importance of considering wave variations in future research due to potential significant outcome differences.

Robust data normalization and analysis are pivotal in biomedical research to ensure that observed differences in populations are directly attributable to the target variable, rather than disparities between control and study groups. ArsHive addresses this challenge using advanced algorithms to normalize populations (e.g., control and study groups) and perform statistical evaluations between demographic, clinical, and other variables within biomedical datasets, resulting in more balanced and unbiased analyses. The tool’s functionality extends to comprehensive data reporting, which elucidates the effects of data processing, while maintaining dataset integrity. Additionally, ArsHive is complemented by A.D.A. (Autonomous Digital Assistant), which employs OpenAI’s GPT-4 model to assist researchers with inquiries, enhancing the decision-making process. In this proof-of-concept study, we tested ArsHive on three different datasets derived from proprietary data, demonstrating its effectiveness in managing complex clinical and therapeutic information and highlighting its versatility for diverse research fields.

Background: Despite advanced analytical methods and increasing data availability, most intensive care unit (ICU) prediction models rely on static measurements. However, longitudinal monitoring of biomarkers may better capture disease progression and support timely, individualized interventions within the framework of predictive, preventive, and personalized medicine (PPPM). Since the COVID-19 pandemic, interest in both static and dynamic modelling has expanded. Therefore, this review aimed to summarize current evidence on the use of longitudinal blood biomarker data in ICU prediction models, assess how the pandemic shaped this research, and report validation strategies. Methods: This scoping review followed the PRISMA-ScR guidelines. PubMed and Google Scholar were searched for studies on blood biomarker trajectory analysis in the ICU published between 2014 and 2025, covering five years before and after the onset of the COVID-19 pandemic. Results: Forty-seven studies were included, mainly from North America (47%), Europe (45%), and Asia (34%). ICU and hospital mortality were the predominant outcomes. Although 53% of studies used pre-pandemic data, 94% were published afterwards. The most frequent biomarker categories were immune response (74.5%) and metabolic/organ function (66.0%). Common biomarkers included platelets and lactate ( n = 9), lymphocytes and mHLA-DR ( n = 6), and creatinine and interleukin-6 ( n = 5). Modelling approaches integrated longitudinal regression-based models (31.9%), latent class-based models (44.7%), and machine-learning/data-driven clustering (27.7%). Trajectory patterns varied depending on both biomarker type and modelling technique. Cox regression, Kaplan-Meier, and logistic regression were commonly applied to assess associations with outcomes. Notably, only 21% of studies reported any form of validation, highlighting a major limitation for clinical applicability. Conclusion: Blood biomarker trajectories have potential to improve dynamic risk prediction and stratification, supporting targeted prevention through early identification of high-risk patterns, and enable more personalized treatment via adaptive, patient-specific approaches. Nevertheless, substantial methodological heterogeneity and the low proportion of validated models limit clinical applicability. Greater standardization and robust validation are essential to facilitate translation into PPPM-oriented intensive care.

Identifying high-risk patients, particularly in intensive care units (ICUs), enhances treatment and reduces severe outcomes. Since the pandemic, numerous studies have examined COVID-19 patient profiles and factors linked to increased mortality. Despite six pandemic waves, to the best of our knowledge, there is no extensive comparative analysis of patients’ characteristics across these waves in Portugal. Thus, we aimed to analyze the demographic and clinical features of 1041 COVID-19 patients admitted to an ICU and their relationship with the different SARS-Cov-2 variants in Portugal. Additionally, we conducted an in-depth examination of factors contributing to early and late mortality by analyzing clinical data and laboratory results from the first 72 h of ICU admission. Our findings revealed a notable decline in ICU admissions due to COVID-19, with the highest mortality rates observed during the second and third waves. Furthermore, immunization could have significantly contributed to the reduction in the median age of ICU-admitted patients and the severity of their conditions. The factors contributing to early and late mortality differed. Age, wave number, D-dimers, and procalcitonin were independently associated with the risk of early death. As a measure of discriminative power for the derived multivariable model, an AUC of 0.825 (p < 0.001; 95% CI, 0.719–0.931) was obtained. For late mortality, a model incorporating age, wave number, hematologic cancer, C-reactive protein, lactate dehydrogenase, and platelet counts resulted in an AUC of 0.795 (p < 0.001; 95% CI, 0.759–0.831). These findings underscore the importance of conducting comprehensive analyses across pandemic waves to better understand the dynamics of COVID-19.