A whole-exome capture and next-generation sequencing was applied to an 11 y/o patient with a clinical history of congenital hypotonia, generalized motor and cognitive neurodevelopmental delay, and severe cognitive deficit, and without any identifiable Syndromic pattern, and to her parents, we disclosed a de novo heterozygous pathogenic mutation, c.697_699del p.Phe233del (rs786204835)(ACMG classification PS2, PM1, PM2, PP5), harbored in the PURA gene (MIM*600473) (5q31.3), associated with Autosomal Dominant Mental Retardation 31 (MIM # 616158). We used the significant improvement in the accuracy of protein structure prediction recently implemented in AlphaFold that incorporates novel neural network architectures and training procedures based on the evolutionary, physical, and geometric constraints of protein structures. The wild-type (WT) sequence and the mutated sequence, missing the Phe233, were reconstructed. The predicted local Distance Difference Test (lDDT) for the PURAwt and the PURA–Phe233del showed that the occurrence of the Phe233del affects between 220–320 amino acids. The distortion in the PURA structural conformation in the ~5 Å surrounding area after the p.Phe233del produces a conspicuous disruption of the repeat III, where the DNA and RNA helix unwinding capability occurs. PURA Protein–DNA docking corroborated these results in an in silico analysis that showed a loss of the contact of the PURA–Phe233del III repeat domain model with the DNA. Together, (i) the energetic and stereochemical, (ii) the hydropathic indexes and polarity surfaces, and (iii) the hybrid Quantum Mechanics–Molecular Mechanics (QM–MM) analyses of the PURA molecular models demarcate, at the atomic resolution, the specific surrounding region affected by these mutations and pave the way for future cell-based functional analysis. To the best of our knowledge, this is the first report of a de novo mutation underpinning a PURA syndrome in a Latin American patient and highlights the importance of predicting the molecular effects in protein structure using artificial intelligence algorithms and molecular and atomic resolution stereochemical analyses.

In critically ill patients, identification of the pathogen may allow for the timely adjustment of antibiotics and improved outcomes. Background/Objectives: The aim of the study was to assess whether performing a multiplex PCR pneumonia panel (PN-panel) in patients with pneumonia in the intensive care unit (ICU) had any effect on mortality or other important clinical outcomes. Methods: A retrospective cohort study was conducted on adult patients with pneumonia who required ICU admission in four institutions in Bogotá between November 2019 and June 2023. Mortality at 30 days, the length of the hospital and ICU stay, the duration of antibiotics, and their association with the PN-panel performance were evaluated using an inverse probability of the treatment weighting to adjust for covariates and potential confounders. Results: A total of 304 patients were included, including 150 with PN-panel, with a mean age of 65.0 years (SD 14.6). SARS-CoV-2 was the primary etiologic agent in 186 (61.2%) patients, and 256 (84.2%) patients had community-acquired pneumonia. No association was found between 30-day mortality and the PN-panel, with a HR of 1.14 (CI 95% 0.76–1.70), although the assessment by an infectious disease specialist was associated with a lower mortality HR of 0.29 (CI 95% 0.19–0.45). There was no association between the PN-panel and antimicrobial therapy duration or other clinical outcomes. Conclusions: The use of the PN-panel was not associated with changes in mortality, the duration of antibiotics, or hospital and ICU stays. To acquire greater rational decision-making, microbiological data produced by this test should be interpreted with aid of an antimicrobial stewardship program oriented by an infectious disease team that could take the clinical data and integrate the information provided.

Background: Hemoglobin (Hb) and hematocrit (Hct) reference intervals (RIs) are critical for diagnosing hematological disorders. However, existing reference values often do not account for demographic and environmental variability. Particularly in countries with altitude gradients, such as Colombia, the absence of locally adjusted intervals may lead to the misclassification of anemia and polycythemia. Therefore, this study aims to establish sex-, age-, and altitude-specific reference intervals for Hb and Hct within the Colombian adult population via an indirect, big-data-based methodology. Methods: This retrospective cross-sectional study used 3.1 million Hb and Hct test results nationwide between 2022 and 2024. After applying the exclusion criteria, Hb data from 667,857 individuals and Hct data from 662,024 individuals were included. The population was stratified by sex, age, and altitude into <1100 m above sea level (m.a.s.l.), 1100–2000 m.a.s.l., and 2000–3000 m.a.s.l. Reference intervals (RIs) were estimated via the refineR algorithm, and the results were compared across altitude categories and against World Health Organization (WHO) anemia and polycythemia thresholds. Results: Hb and Hct concentrations increased with altitude in all sexes and age groups. Compared with women, men presented higher mean values and narrower RIs, whereas older adults presented greater variability. Compared with WHO thresholds, a significant proportion of individuals living above 2000 m exceeded polycythemia cutoffs without clinical evidence of disease, suggesting the need for altitude-adjusted diagnostic criteria. Conclusions: This study provides the first large-scale, data-driven reference intervals for Hb and Hct in Colombia, adjusted for altitude, sex, and age. The implementation of locally derived RIs may improve diagnostic accuracy and prevent the over- or underdiagnosis of hematological disorders, with direct implications for clinical decision-making and public health policy.

A whole-exome capture and next-generation sequencing applied to an 11 y/o patient with a clinical history of congenital hypotonia, generalized motor and cognitive neurodevelopmental delay, severe cognitive deficit, without any identifiable Syndromic pattern, and to her parents, disclosed a de novo heterozygous pathogenic mutation, c.697_699del p.Phe233del (rs786204835)(ACMG classification PS2, PM1, PM2, PP5), harbored in the PURA gene (MIM*600473) (5q31.3), associated to Autosomal Dominant Mental Retardation 31 (MIM # 616158). We used the significant improvement in the accuracy of protein structure prediction recently implemented in AlphaFold that incorporates novel neural network architectures and training procedures based on the evolutionary, physical, and geometric constraints of protein structures. The wild-type (WT) sequence and the mutated one, missing the Phe233, were reconstructed. The predicted local Distance Difference Test (lDDT) for the PURA WT and the PURA-Phe233del showed that the occurrence of the Phe233del affects between 220-320 amino acids. The distortion in the PURA structural confor-mation in the ~5Å surrounding area after the p.Phe233del produces a conspicuous disruption of the repeat III, where the DNA and RNA helix unwinding capability occurs. PURA Protein-DNA Docking corroborated these results in silico Analysis that showed a loss of the contact of the PURA-Phe233del III repeat domain model with the DNA. Together, i) the energetic and stereochemical, ii) the hydropathic indices and polarity surfaces, and iii) the hybrid Quantum Mechanics-Molecular Mechanics (QM-MM) analyses of the PURA molecular models demarcate at the atomic resolution the specific surrounding region affected by these mutations and paves the way for future cell-based functional analysis. To the best of our knowledge, this is the first report of a de novo mutation underpinning a PURA syndrome in a Latin American patient and highlights the importance of predicting the molecular effects in protein structure using artificial intelligence algorithms and molecular and atomic resolution stereochemical analyses.