Some important aspects of implementing tandem mass spectrometry in a routine clinical laboratory environment

Tijekom posljednjih godina kombinacija tekućinske kromatografije visoke učinkovitosti (engl. high performance liquid chromatography, HPLC) i dvojne spektrometrije masa (engl. tandem mass spectrometry, MS/MS) (kombinacija koja je također poznata kao HPLC-MS/MS), postala je pouzdani analitički postupak. Njena primjena u terapijskom praćenju koncentracije lijeka (engl. therapeutic drug monitoring, TDM) pokazala se boljom od uobičajeno primjenjivanih imunokemijskih analiza. Ta je tehnika postala obveznim pomagalom u mnogim kliničkim laboratorijima, posebice u kontekstu praćenja koncentracije imunosupresijskih lijekova gdje se smatra "zlatnim" standardom. Međutim, postavljanje uređaja za HPLC-MS/MS je zahtjevno u smislu validacije analiza te robusnosti mjerenja. U ovom se pregledu daje gruba smjernica kao pomoć u procesu uspostave takvih uređaja u rutinskim kliničkim uvjetima.During past years, the combination of high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS), also known as HPLC-MS/MS, matured to reliable analytical instrumentation. Its application in therapeutic drug monitoring (TDM) has been shown to be superior to normally applied immunoassays. Especially in the context of immunosuppressant medication level surveillance, this technique has become an indispensable tool in many clinical laboratories and is considered the gold standard. However, setting up a HPLC-MS/MS platform is demanding in terms of assay validation and the robustness of testing. This review is to provide a rough guideline aiding the implementation process of such a platform into a routine clinical environment

Immunoglobulin subclass 4 for the diagnosis of immunoglobulin subclass 4-associated diseases in an unselected liver and pancreas clinic population

AbstractAimsThe diagnosis of autoimmune pancreatitis (AIP) and immunoglobulin subclass 4 (IgG4)-associated cholangitis (IAC) is based on imaging studies, serology, histology and a response to steroid therapy. The major serological finding is an elevation of the serum IgG4 concentration. Previous studies have shown that its sensitivity is about 70% and its specificity exceeds 90% at a cut-off of 140mg/dl in selected patient populations. The aim of the present study was to assess the performance of serum IgG4 as a diagnostic parameter in an unselected liver and pancreas clinic population.Methods and resultsIgG4 was prospectively determined in 1412 patients and clinical diagnoses were recorded from a review of patient charts. The prevalence of AIP or IAC in the entire cohort was 1.1% (n= 15). The sensitivity of IgG4 for the diagnosis of AIP and IAC was 80% and the specificity was 86% at a cut-off value of ≥135mg/dl. The positive predictive value and the negative predictive value were 6% and 99.7%, respectively. The most common differential diagnosis in patients with elevated IgG4 was liver cirrhosis.ConclusionIgG4 has a reasonable sensitivity and specificity in a liver and pancreas clinic population, where liver cirrhosis appears to be the most frequent differential diagnosis for elevated IgG4 concentrations

Testosterone Deficiency Is a Risk Factor for Severe COVID-19

BackgroundMale sex is related to increased COVID-19 severity and fatality although confirmed infections are similarly distributed between men and women. The aim of this retrospective analysis was to investigate the impact of sex hormones on disease progression and immune activation in men with COVID-19.Patients and MethodsWe studied for effects of sex hormones on disease severity and immune activation in 377 patients (230 men, 147 women) with PCR-confirmed SARS-CoV-2 infections hospitalized at the Innsbruck University Hospital between February and December 2020.ResultsMen had more severe COVID-19 with concomitant higher immune system activation upon hospital admission when compared to women. Men with a severe course of infection had lower serum total testosterone (tT) levels whereas luteinizing hormone (LH) and estradiol (E2) levels were within the normal range. tT deficiency was associated with elevated CRP (rs = - 0.567, p < 0.001), IL-6 levels (rs = - 0.563, p < 0.001), lower cholesterol levels (rs = 0.407, p < 0.001) and an increased morbidity and mortality. Men with tT levels < 100 ng/dL had a more than eighteen-fold higher in-hospital mortality risk (OR 18.243 [95%CI 2.301 – 144.639], p = 0.006) compared to men with tT levels > 230 ng/dL. Moreover, while morbidity and mortality showed a positive correlation with E2 levels at admission, we detected a negative correlation with the tT/E2 ratio upon hospital admission.ConclusionHospitalized men with COVID-19 present with rather low testosterone levels linked to more advanced immune activation, severe clinical manifestations translating into an increased risk for ICU admission or death. The underlying mechanisms remain elusive but may include infection driven hypogonadism as well as inflammation mediated cholesterol reduction causing gonadotropin suppression and impaired androgen formation. Finally, in elderly late onset hypogonadism might also contribute to lower testosterone levels

Complexity of coronary artery disease and the release of cardiac biomarkers after CABG

Objective: In patients with complex coronary artery disease (CAD) undergoing cardiac surgery, myocardial protection might be impaired due to microvascular obstruction, resulting in myocardial injury and subsequent biomarker release. Therefore, this study investigated the correlation between the complexity of CAD, reflected by the SYNTAX Score, and the release of cardiac biomarkers after CABG. Methods: In a consecutive series of 919 patients undergoing isolated CABG SYNTAX scores I and II were calculated to assess the complexity of CAD. Levels of high sensitivity cardiac troponin T (hs-cTnT) and creatine kinase-myocardial band (CK-MB) were routinely measured once before and serially after surgery. Patients were divided into tertiles according to their SYNTAX Scores I and II. Spearman correlations and regression models were performed to measure the degree of association between the release of hs-cTnT and CK-MB and the SYNTAX Scores. Results: Patients with a higher SYNTAX Score I had more comorbidities reflected in a higher EuroSCORE II. Preoperatively, higher levels of cardiac biomarkers were found in patients with higher SYNTAX Score II. No correlation was observed between hs-cTnT, CK-MB and SYNTAX Score I or II. Regression models did not show any association between cardiac biomarkers and the complexity of CAD. Conclusion: The complexity of CAD is not associated with the release of cardiac biomarkers after CABG. Factors influencing postoperative biomarker release need to be elucidated in future trials to include postoperative biomarker release into risk stratification models predicting outcome after cardiac surgery

Plasma homocysteine levels and associated factors in community-dwelling adolescents: the EVA-TYROL study

BackgroundHomocysteine (Hcy) has been associated with an adverse cardiovascular risk profile in adolescents. Assessment of the association between plasma Hcy levels and clinical/laboratory factors might improve our understanding of the pathogenesis of cardiovascular disease.MethodsHcy was measured in 1,900 14- to 19-year-old participants of prospective population-based EVA-TYROL Study (44.3% males, mean age 16.4 years) between 2015 and 2018. Factors associated with Hcy were assessed by physical examination, standardized interviews, and fasting blood analysis.ResultsMean plasma Hcy was 11.3 ± 4.5 µmol/L. Distribution of Hcy was characterized by extreme right skew. Males exhibited higher Hcy and sex differences increased with increasing age. Univariate associations with Hcy emerged for age, sex, body mass index, high-density lipoprotein cholesterol, and for factors pertaining to blood pressure, glucose metabolism, renal function, and diet quality, whereas the most important multivariate predictors of Hcy were sex and creatinine.DiscussionClinical and laboratory factors associated with Hcy in adolescents were manifold, with sex and high creatinine identified as strongest independent determinants. These results may aid when interpreting future studies investigating the vascular risk of homocysteine

In Vitro Effects of Hypoxia and Reoxygenation on Human Umbilical Endothelial Cells

Peer Reviewe

Clonal Hematopoiesis of Indeterminate Potential: A Multidisciplinary Challenge in Personalized Hematology

Clonal hematopoiesis of indeterminate potential (CHIP) is a common age-related condition that represents a potential pre-phase of hematologic neoplasm. Next-generation sequencing (NGS) is used to detect and monitor clonal hematopoiesis, and the spectrum of mutations substantially overlaps with that of myeloid neoplasms with DNMT3A, TET2, ASXL1, and JAK2 being the most frequently mutated. While, in general, the risk of progression to an overt myeloid neoplasm is only modest, the progression risk increases in patients with unexplained cytopenia or multiple mutations. In addition, CHIP represents a previously unrecognized major risk factor for atherosclerosis and cardiovascular disease (CVD), including coronary heart disease, degenerative aortic valve stenosis, and chronic heart failure; and a causative role of CHIP in the development of CVD has been demonstrated in vitro and in vivo. The management of patients with CHIP is a rapidly emerging topic in personalized medicine, as NGS has become widely available for clinical medicine. It requires a highly multidisciplinary setting, including hematology/oncology, cardiology, (clinical) pathology, and genetics for individualized guidance. Further research is urgently needed to provide robust evidence for future guidelines and recommendations on the management of patients with CHIP in the era of personalized medicine

Impact of a single, short morning bright light exposure on tryptophan pathways and visuo- and sensorimotor performance: a crossover study

Abstract Background Bright light (BL) has been shown to be effective in enhancing both cognitive and physical performances. Alterations in nighttime melatonin levels have also been observed. However, evaluations of light-induced changes in the preceding biochemical processes are absent. Therefore, the impact of a single morning BL exposure on sensorimotor and visuomotor performance, as well as tryptophan (trp) and trp metabolites, was evaluated in this study. Methods In a crossover design, 33 healthy volunteers were randomly exposed to 30 min of < 150 lx at eye level (office light, OL) and 5000 lx at eye level (bright light, BL) of 6500 K in the morning hours. Trp, sulfatoxymelatonin (aMT6s), and kynurenine (kyn) courses over the morning hours were analyzed, and changes in sensori- and visuomotor measures were examined. Results Motoric performance increased in both setups, independent of light intensity. aMT6s and kyn decreased equally under both lighting conditions. Trp levels decreased from a mean (95% confidence interval) of 82.0 (77.2–86.9) to 66.5 (62.5–70.1) in the OL setup only. Conclusion These data suggest that BL in the morning hours has a limited effect on visuo- and sensorimotor performance. Nevertheless, trp degradation pathways in the morning show diverse courses after OL and BL exposure. This suggests that trp courses can potentially be altered by BL exposure