ABSTRACT
Kryptor system was proven to be a rapid, standard method for pregnancy-associated plasma protein A and proform eosinophilic major basic protein (PAPP-A/proMBP) complex detection in coronary artery disease (CAD). No age and/or gender differences in 51 controls and 110 stable coronary artery disease (SCAD) patients were found. SCAD patients did not differ from controls and no difference in PAPP-A/proMBP levels with regards to the number of affected vessels was found. In 21 unstable angina pectoris (UAP), in 35 without and 66 with ST elevation acute myocardial infarctions (NSTEMI, STEMI respectively) patients PAPP-A/proMBP levels were increased (P=0.004 and P<0.0005, respectively). PAPP-A/proMBP levels did not correlate with cardiac troponin I (cTnI) in STEMI and NSTEMI patients. PAPP-A/ proMBP increase was more frequent than cTnI (P=0.036) within the early phase of STEMI. In NSTEMI patients PAPP-A/proMBP positivity was present in 50% of cTnI negative cases. Receiver operating characteristic (ROC) analysis revealed the highest diagnostic accuracy of PAPP-A/proMBP (0.919) in STEMI cTnI positive cases. The highest specificity/sensitivity PAPP-A/proMBP levels for particular acute coronary syndrome (ACS) types were 10.65-14.75 mIU/l. Combination of PAPP-A/proMBP with cTnI increases their diagnostic efficacy within the early phase of ACS. Our results suggest that PAPP-A/proMBP complex is involved in processes preceding vulnerable plaque development in ACS.
Subject(s)
Acute Coronary Syndrome/diagnosis , Coronary Artery Disease/diagnosis , Eosinophil Major Basic Protein/analysis , Pregnancy-Associated Plasma Protein-A/analysis , Acute Coronary Syndrome/blood , Adult , Age Factors , Aged , Aged, 80 and over , Biomarkers/blood , Case-Control Studies , Cohort Studies , Coronary Artery Disease/blood , Coronary Artery Disease/classification , Enzyme-Linked Immunosorbent Assay , Eosinophil Major Basic Protein/metabolism , Female , Humans , Immunoassay/instrumentation , Immunoassay/methods , Male , Middle Aged , Pregnancy-Associated Plasma Protein-A/metabolism , Protein Precursors/analysis , Protein Precursors/metabolism , ROC Curve , Reference Values , Risk Assessment , Sensitivity and Specificity , Sex Factors , Statistics, Nonparametric , Troponin I/analysis , Troponin I/metabolismABSTRACT
OBJECTIVE: Determination of the sex of the foetus from enriched nuclear red blood cells (NRBC) circulating in maternal blood during pregnancy. METHODS: NRBC were enriched from 13-28 ml peripheral blood of 32 pregnant women using double MACS procedure. NRBCs were enriched by magnetic activated cell sorting using anti-CD71 (transferrin receptor) monoclonal antibodies. Unwanted leukocytes were depleted using monoclonal antibodies against CD14 and CD45. The sex of the foetus was analysed by using dual-colour FISH with X and Y specific probes. The experimental results obtained from the noninvasive procedure were compared to karyotype obtained from amniocentesis or chorionic villus sampling. RESULTS: In 15 out of 17 male foetuses we could identify one X and one Y signal. In another 15 pregnant women carrying female foetuses two X signals were observed. CONCLUSION: NRBC circulating in blood of pregnant women can be used as an alternative source for determination of the sex of the foetus with a risk of false negative results (2/17, 12%). The problem of false negative results can be solved by using more sophisticated methods of enrichment and preparedness of the slides for FISH analysis.
Subject(s)
Erythroblasts , Pregnancy/blood , Sex Determination Analysis/methods , DNA Probes , Female , Humans , In Situ Hybridization, Fluorescence , MaleABSTRACT
BACKGROUND: Acute graft versus host disease (GvHD) remains a severe complication of allogeneic haematopoietic stem cell transplantation (HSCT). Our study summaries results of skin explant assay (SEA) as a pretransplant GvHD predictive test in a cohort of paediatric (n = 33) and adult (a = 8) patients receiving grafts from their HLA identical siblings (n = 28), haploidentical relatives (n = 3) and unrelated donors (n = 10). Results GvHD prediction are correlated with the occurrence and severity of acute GvHD posttransplant and effect of GvHD prophylaxis on GvHD clinical outcome is evaluated. METHODS AND RESULTS: SEA utilises responding lymphocytes of the donor, which are sensitized firstly in vitro by mononuclears cells of patient in allogeneic mixed lymphocyte culture (MLC) and subsequently co-cultured with recipient's skin. Histopathological changes found in patients' skin explants are evaluated according to standard Lerner classification for acute GvHD. In general, GvHD predictive results in SEA correlated with GvHd clinical outcome in 28 out of 41 tested patients (68%, p = 0.015). In a cohort of HLA identical sibling transplants GvHD predictive results correlated with clinical manifestation of acute GvHD only in 15 out of 28 patients on individual GvHD prophylaxis. GvHD prophylaxis in the form of cyclosporine A (CsA) combined with short-term methotrexate (MTX) reduced the risk of acute GvHD in 10 out of 14 transplanted patients (71%) meanwhile CsA alone prophylaxis only in 1 out of 5 patients (20%). In a cohort of unrelated pairs on CsA/MTX prophylaxis combined with horse anti-lymphocyte globuline (ALG) correlated the GvHD prediction with GvHD clinical outcome (100%, p = 0.003). In all patients transplanted with the grafts from their haploidentical relatives the occurrence of severe GvHD was predicted. CONCLUSION: Skin explant assay helps identify pretransplant patients at higher risk of severe acute GvHD. GvHD predictive results enable the transplantation team to individualise GvHD prophylaxis and to optimise selection of the donor.