A Search for New Biological Pathways in Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy by Proteomic Research.

Background/Objectives: Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is a hereditary small vessel disease leading to significant morbidity and mortality. Despite advances in genetic diagnosis, the underlying pathophysiology remains incompletely understood. Proteomic studies offer insights into disease mechanisms by identifying altered protein expression patterns. Here, we conducted a proteomic analysis to elucidate molecular pathways associated with CADASIL. Methods: We enrolled genetically diagnosed CADASIL patients and healthy, genetically related controls. Plasma samples were subjected to proteomic analysis using the Olink platform, measuring 552 proteins across six panels. The data were analyzed from several approaches by using three different statistical methods: Exploratory Principal Component Analysis (PCA) and Partial Least Squares-Discriminant Analysis (PLS-DA), differential expression with moderated t-test, and gene set enrichment analysis (GSEA). In addition, bioinformatics analysis, including volcano plot, heatmap, and Variable Importance on Projection (VIP) scores from the PLS-DA model were drawn. Results: Significant differences in protein expression were observed between CADASIL patients and controls. RSPO1 and FGF-19 exhibited elevated levels (p < 0.05), while PPY showed downregulation (p < 0.05) in CADASIL patients, suggesting their involvement in disease pathogenesis. Furthermore, MIC-A/B expression varied significantly between patients with mutations in exon 4 versus exon 11 of the NOTCH3 gene (p < 0.05), highlighting potential immunological mechanisms underlying CADASIL. We identified altered pathways using GSEA, applied after ranking the study data. Conclusions: Our study provides novel insights into the proteomic profile of CADASIL, identifying dysregulated proteins associated with vascular pathology, metabolic dysregulation, and immune activation. These findings contribute to a deeper understanding of CADASIL pathophysiology and may inform the development of targeted therapeutic strategies. Further research is warranted to validate these biomarkers and elucidate their functional roles in disease progression.

Longitudinal changes of angiogenic factors as a potential predictive tool in women with suspected preeclampsia.

OBJECTIVES: To investigate whether longitudinal changes of angiogenic factors (AF) sFlt-1, PlGF, and the sFlt-1/PlGF ratio, measured following identification of symptoms of preeclampsia (PE), could provide complementary information to the isolated measurements used in current clinical practice. STUDY DESIGN: Retrospective observational study. Sixty women with suspected PE and two AF results measured before gestational week (GW) 34 were included. Daily variation (DV) of AF was calculated from delta values and days elapsed between measurements. Through ROC analysis, the predictive performance of DV for PE-related events was estimated. Kaplan-Meier survival curves resulting from applying cutoff values were assessed. RESULTS: The sFlt-1, PlGF, and sFlt-1/PlGF ratio baseline levels showed significant differences between women without PE and women who developed early-onset PE (P < 0.001). DV of sFlt-1 and sFlt-1/PlGF ratio increased according to the severity of PE, showing significant differences in both pairs of groups compared (p < 0.001), so they were selected as potential predictors. Higher AUC values resulting from ROC analysis were 0.78 for early-onset PE, 0.88 for early-onset severe PE, 0.79 for occurrence of adverse maternal outcomes, and 0.89 for delivery before 37 GW, with sensitivity and specificity values higher than 0.71 and 0.80, respectively. The Kaplan-Meier analysis yielded significantly different curves (log-rank < 0.05), with shorter time-to-delivery as DV increased. CONCLUSION: Our results support the existence of a correlation between a progressive PlGF and sFlt-1 imbalance and a more aggressive clinical course of PE, detectable from the finding of PE symptoms. Its monitoring could be a useful predictive tool in women with suspected PE.

Evaluation of the Premier Hb9210 instrument for HbA1c determination.

Background: Glycated hemoglobin measurements are a valuable tool for long-term blood glucose monitoring and the diagnosis of diabetes. Its widespread use has been made possible due to the development of new analytical methods with improved performances and standardization with reference materials. The aim of the present study was to evaluate the Trinity Biotech Premier Hb9210 analyzer for the measurement of HbA1c.Methods: The precision was assessed using the CLSI EP-15A3 and EP-10A3 protocols. The latter was also used to investigate linearity, carryover, and linear drift. The comparison study was performed between Premier Hb910 and Tosoh HLC-723 G8 through Passing-Bablok regression and the Bland-Altman plot. The Fleiss Kappa index was used to assess the degree of agreement. The interference of Hb variants was investigated using samples with Hb variants S, C, D, E, J, and Seville.Results: Within-run and between-run imprecision fell between 0.37% and 1.16%. No statistically significant nonlinearity, carry-over, and/or drift were observed. The resulting regression line of the Passing-Bablok analysis was y = 0.00 + 1.00x. The Pearson correlation coefficient was 0.997. In the Bland-Altman plot, the relative bias was 0.01%. The overall Fleiss Kappa index was 0.9. No interference from hemoglobin variants was observed.Conclusion: The Premier Hb9210 demonstrated a high degree of automation, reproducibility, good agreement, minimal carry-over effect, and excellent linearity across the wide range of HbA1c levels commonly found in diabetic patients and was not influenced by Hb variants.

Premier Hb9210 can be used as an alternative to monitor glycemic status.Premier Hb9210 is not affected by common hemoglobin variants.Premier Hb9210 can correctly classify diabetic patients.

LDH/ADA ratio in pleural fluid for the diagnosis of infectious pleurisy.

Pleural effusion (PE) is a common medical concern, often requiring thoracentesis for a definitive diagnosis. An elevated pleural fluid adenosine deaminase (ADA) may indicate tuberculosis, but this is not always the case. This study aimed to evaluate the accuracy of biomarkers determined in pleural fluid and propose a new diagnostic strategy for PE in patients with high levels of ADA in pleural fluid. This retrospective analysis studied patients with PE who received thoracentesis for the first time with an ADA level of > 33 U/L in the pleural fluid analysis at two tertiary hospitals from March 2019 to March 2023. Demographic and clinical data, as well as pleural fluid biomarkers and their ratios, were studied and compared between different PE groups, and a decision tree was developed. During the study period, 259 patients were enrolled, with four different types of PE: parapneumonic (PPE) 155, tuberculosis (TPE) 41, malignant (MPE) 50, and miscellaneous 13. Biomarkers and their ratios performed well in the differential diagnosis of PE, with the LDH/ADA ratio distinguishing between PPE and non-PPE with sensitivity and specificity of 98.06% and 98.08%, respectively. The combination of LDH/ADA ratio, ADA, and mononuclear cell percentage was identified as important factors for creating a decision tree with an overall accuracy of 89.96%. The pleural fluid LDH/ADA ratio was a useful diagnostic for distinguishing PPE from non-PPE, and a decision tree with an accuracy of 89.96% was created to differentiate the four forms of PE in clinical situations.

Long-term follow-up with a clinical decision support system based on laboratory reports to manage patients with biochemical recurrence after radical prostatectomy.

INTRODUCTION: Currently, prostate cancer (PCa) is the second most common cause of cancer death, and radical prostatectomy (RP) remains the primary treatment for localized PCa. Although there is no consensus on an optimal strategy, the determination of total serum prostate-specific antigen (tPSA) is the cornerstone for the detection of postoperative biochemical recurrence (BCR). The aim of this study was to evaluate the prognostic utility of serial tPSA levels together with other clinicopathological factors and to assess the impact of a commentary algorithm implemented in our laboratory information system. METHODS: A descriptive and retrospective study of patients with clinically localized PCa who underwent RP. BCR-free survival was calculated over time (Kaplan-Meier analysis), and the ability of different clinicopathological factors to predict BCR was studied (univariate and multivariate analyses) with Cox models. RESULTS: A total of 203 patients underwent RP, of whom 51 presented with BCR during follow-up. In the multivariate model, doubling of tPSA, the Gleason score, tumour stage and tPSA nadir were detected as independent predictors of BCR. CONCLUSION: A patient with undetectable tPSA after 1959 days of RP is unlikely to develop BCR, regardless of preoperative or pathologic risk factors. Furthermore, doubling of tPSA in the first 2 years of follow-up was the main prognostic factor for BCR in patients undergoing RP. Other prognostic factors included a tPSA nadir detectable after surgery, a Gleason score ≥ 7 and a tumour stage T ≥ 2c.

Evaluation of the Congenital Hypothyroidism Detection Strategy in Extremely Preterm Infants in Western Andalusia.

OBJECTIVE: This study aimed to identify extremely premature infants (< 31 weeks of gestation and/or <1,500 g) affected by congenital hypothyroidism (CH) with delayed elevation of thyrotropin (TSH) and to evaluate the detection strategy for this pathology in our reference screening population. STUDY DESIGN: A descriptive and retrospective study was carried out with samples collected from western Andalusia and the autonomous city of Ceuta. RESULTS: This protocol allowed us to detect six neonates with delayed TSH elevation. One of them, due to serious heart problems, died without being able to confirm CH. In two neonates, however, it was possible to detect CH, another two presented a persistent TSH elevation but normal free T4, and another one presented a temporary TSH elevation. CONCLUSION: It is essential to repeat the CH screening in extremely premature infants, not only at the age of 15 days but also with a third sample at the moment of hospital discharge to detect cases with delayed TSH elevation. KEY POINTS: · The Newborn Screening Programs are an essential activity of preventive medicine.. · Extremely preterm infants have a very high risk of CH.. · Optimal management of thyroid dysfunction in this population remains to be established..

ClotPro Viscoelastometry Evaluation in Cardiac Surgery With Cardiopulmonary Bypass.

OBJECTIVE: The aim of this study was to assess the reproducibility of the new viscoelastic analyzer ClotPro, and compare the parameters this system produces with the ROTEM delta system in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB). DESIGN: A prospective and observational study. SETTING: At a university hospital. PARTICIPANTS: Forty adult patients undergoing cardiac surgery with CPB. INTERVENTIONS: Correlations were calculated between ClotPro and ROTEM delta parameters. MEASUREMENTS AND MAIN RESULTS: The ClotPro showed a high reproducibility in most of the parameters of each test; whereas ROTEM delta, although showing a low coefficient of variation in the parameters related to clot firmness, showed a high variability in the coagulation times. Excellent correlations were observed in most of the parameters of each test between ROTEM delta and ClotPro (≥0.93). However, a moderate correlation was obtained between the clotting time of the EXTEM and the EX-test (0.54). The concordance of amplitudes at different times within each test was almost 100% on both thromboelastometers. Regarding absolute differences in the test results, most of the measurements showed significant differences (p < 0.0001) between both devices. CONCLUSIONS: ClotPro can be used as an alternative to ROTEM delta to evaluate coagulation function in cardiac surgery, but specific reference ranges need to be established first.

Neonatal Screening for Sickle Cell Disease in Western Andalusia: Results and Lessons Learnt after 3 Years of Implementation.

OBJECTIVE: The aim of this study was to present the results obtained in the Newborn Screening Program (NSP) for sickle cell disease (SCD) in western Andalusia and the autonomous city of Ceuta in the first 3 years of implementation, and to describe the discrepancies found in the diagnosis of hemoglobinopathies between the screening method and the confirmatory tests. STUDY DESIGN: A descriptive and retrospective study was carried out, and the findings obtained in the newborns included in the NSP between November 2018 and December 2021 were analyzed. RESULTS: A total of 111,205 samples were screened by high-performance liquid chromatography (HPLC). The birth prevalence of SCD, sickle cell trait, hemoglobin C carriers, and the compound heterozygosity Hb C/ß-thalassemia was 1/12,356, 1/467, 1/1,278, and 1/55,602 newborns, respectively. Although there was a correlation between the first-line HPLC screening technique (VARIANTnbs HPLC analyzer, Bio-Rad) and the confirmatory tests in most cases, major discrepancies were found in detecting carriers of G-Philadelphia, D, E, and O-Arab hemoglobin variants, with the former having an incidence of 1/10,110 and the others 1/22,241. The carrier status of Hb G-Philadelphia produced an FAD pattern on the screening method that could be mistaken as Hb D, while Hb O-Arab was identified as an FA5 pattern. Hb D was initially recognized as Hb D in two cases. CONCLUSION: An NSP requires at least two different combined methods in order to identify the hemoglobin variant with sufficient certainty. Furthermore, even though software solutions for HPLC suggest a pattern, it must be confirmed with another technique to obtain a correct interpretation of the chromatograms. KEY POINTS: · The NSPs are an essential activity in preventive medicine.. · At least two different combined methods are required to correctly identify hemoglobin variants.. · Different variants can produce a similar or identical pattern by a single method..

Evaluation of Routine Blood Tests for Diagnosis of Suspected Coronavirus Disease 2019.

BACKGROUND: Real-time reverse transcription polymerase chain reaction assay (RT-PCR) is the gold standard for diagnosis of coronavirus disease 2019 (COVID-19); however, it is not universally available and may have limitations in response times. The aim was to evaluate the routine blood tests for diagnosis of COVID-19, determining the diagnostic accuracy of blood biomarkers to differentiate between patients with and without COVID-19. METHODS: Clinical charts, nursing records, laboratory findings, and chest x-rays from adult patients with clinical suspicion of COVID-19 (fever, cough and/or dyspnea) at hospital admission were reviewed. Patients were classified into two groups according to RT-PCR COVID-19: positive (COVID-19) or negative (NON-COVID-19). Diagnostic accuracy was determined by analyzing receiver operating characteristic (ROC) curve, calculating the area under the ROC curve (AUC) and the cutoff value. In order to reduce the number of false positives, the cutoff value with a specificity of 80% was considered. RESULTS: Two hundred three patients (101 females, 102 males) with ages between 18 and 96 years (mean = 61.3) were studied. Ninety-four were COVID-19 and 109 were NON-COVID-19. Plasma ferritin level was the most accurate biomarker (AUC = 0.847 and 0.804 in women and men, respectively). The following diagnostic criteria for suspected COVID-19 were established with biomarker cutoff values to differentiate between COVID-19 and NON-COVID-19 patients: lymphocytes ≤ 1.0 x 109/L; eosinophils ≤ 0.02 x 109/L; ferritin > 125% of upper reference limit (URL); LDH > 125% of URL; hsCRP > 80 mg/L; and D-dimer > 1.2 mg/L. Sensitivity was 66%, 64% 62%, 46%, 43%, and 33% for ferritin, eosinophils, LDH, hsCRP, lymphocytes, and D-dimer, respectively. Of those determined to be COVID-19 patients, 91% met one or more of the diagnostic criteria with these blood biomarkers, and of the NON-COVID-19 patients, 47% did not met any diagnostic criteria. CONCLUSIONS: Blood counts of lymphocytes and eosinophils, and plasma levels of D-dimer, LDH, hsCRP, and ferritin can be used to differentiate patients with and without COVID-19 and as a tool for diagnosis of suspected COVID-19 in adult patients at hospital admission.