PAPP-A Concentrations Research Articles

Prenatal Risk Calculation (PRC) 3.0: An Extended DoE-Based First-Trimester Screening Algorithm Allowing For Early Blood Sampling.

Both previous versions of the German PRC algorithm developed by our group for routine first-trimester screening relied on the assumption that maternal blood sampling and fetal ultrasonography are performed at the same visit of a pregnant women. In this paper we present an extension of our method allowing also for constellations where this synchronization is abandoned through preponing blood sampling to dates before 11 weeks of gestation. In contrast to the directly measured concentrations of the serum parameters PAPP-A and free ß-hCG, the logarithmically transformed values could be shown to admit the construction of reference bands covering the whole range from 16 to 84 mm CRL [corresponding to 63 to 98 days of gestation]. Prior to determining reference limits from which the DoEs for each individual patient had to be calculated, the log concentrations of all PAPP-A and free ß-hCG values were transformed once more using the calibration approach established in 1 for the elimination of the influence of maternal weight. Although that part of the database which was available for estimating the reference bands for blood sampling times prior to 11 weeks of gestation was comparatively sparse (898 out of 186 215 pregnancies with euploid outcome), the key statistical characteristics of the extended risk-calculation procedure turned out to be very satisfactory. Using the same cutoff value of 1:150 for the posterior risks of trisomy 21 and 13/18, the overall FPR (false positive rate) for diagnosing a T21 was found to be 3.42%. The corresponding DTR (detection rate) was obtained to be 86.8% and thus exceeded the DTR attained by PRC 2.0 for trisomy 21. For trisomies 13 and 18, the proportions of patients with calculated posterior risks exceeding the cutoff value of 1:150 were obtained to be 1.60% (=FPR) and 86.4% (=DTR). Transforming the measured concentrations of PAPP-A and free ß-hCG to the logarithmic scale allows one to extend the DoE-based algorithm developed by the FMF Germany for diagnosing trisomies 21 and 13/18 in such a way that it can be applied to constellations where blood sampling is done before 11 weeks of gestation.

CLINICAL SIGNIFICANCE OF PROTEIN GROWTH AND INJURY FACTORS IN ACUTE CORONARY SYNDROME

Aim . As fundamentals for current study we layed upon the comparison tasks for concentration of pregnancy associated plasma protein (PAPP-A) and insulin-like growth factor 1 (IGF-1) in blood plasma of patients with acute coronary syndrome (ACS); confirmation of biological role of PAPP-A and IGF-1 as interacting components of intravascular injury and reparation of atherosclerotic plaque; clinical significance of PAPP-A and IGF-1 in ACS patients. Material and methods . We measured concentrations of PAPP-A and IGF-1 in 71 patient with ACS, mean age 57 year old. The study group had a diagnosed myocardial infarction (MI) in 44 patients, and unstable angina in 27; 9 death cases. Synchronically, we measured PAPP-A and IGF-1 in the control and comparison group. Comparison group consisted of 40 arterial hypertension patients and with stable forms of ischemic heart disease. Controls were 20 almost healthy persons. Results . The concentrations of PAPP-A and IGF-1 in ACS patients that were different from controls and comparison. In unstable angina patients PAPP-A is higher than in controls 3,56 times, and in MI 11,6 times. PAPP-A in MI is 3,2 times higher than in unstable angina. In unstable angina the highest found levels of IGF-1 are 1,2 times higher than in controls. The lowest IGF-1 were in lethal cases of MI, and were 1,27 lower than in controls. Conclusion. PAPP-A and IGF-I are new biochemical markers of intravascular injury and reparation of atherosclerotic plaques, and can be applied in urgent cardiology as analyzers of destruction and morphological instability of atherosclerotic plaques in ACS.

First trimester combined screening test in pregnancies derived from blastocyst transfer

ObjectiveTo investigate whether first trimester combined screening for major fetal trisomies is influenced by assisted reproduction techniques from blastocyst transfer, with or without cryopreservation. Study designretrospective case-control analysis involving 298 singleton pregnancies with euploid fetuses recruited between 2012 and 2014. Forty-seven women conceived with fresh blastocysts from in vitro fertilization cycles (fresh-blasto), 51 with frozen–thawed blastocysts (frozen-blasto) and 200 were natural conceptions (controls). All cases underwent ultrasound assessment at 11+0–13+6 weeks with measurements of crown rump length, nuchal translucency, free β-hCG and PAPP-A concentrations. The outcome of pregnancy was recorded (gestational age at birth, birth weight, obstetric complications). ResultsBaseline characteristics and pregnancy outcomes did not differ substantially among the study groups. The median delta nuchal translucency was significantly higher both in frozen-blasto group (median: 0.27mm; Interquartile Range [IQR]: 0.02–0.44; p<0.001) and in fresh-blasto group (median: 0.17mm; IQR: 0.04–0.39; p=0.014) as compared to control group (median: 0.06mm; IQR: −0.08 to 0.20), whereas it was not different in the frozen-blasto compared to the fresh-blasto group. The median free β-hCG multiples of the median (MoMs) was significantly higher both in frozen-blasto group (median 1.15; IQR 0.83–2.11; p=0.001) and in fresh-blasto group (median 0.95; IQR 0.52–1.48; p=0.001) as compared to control group (median 0.99; IQR 0.57–1.26), and it was also higher in frozen-blasto group compared to fresh-blasto group (p<0.001). The three groups showed no significant differences in the median PAPP-A MoMs. The median delta crown rump length was also not significantly different among the three groups. Conclusionsin assisted reproduction technique pregnancies from blastocyst transfer, with or without cryopreservation, both the nuchal translucency measurement and free β-hCG concentration are higher as compared to spontaneous conceptions, whereas PAPP-A does not show any significant difference. These features are apparently unrelated to the outcome of pregnancy and may be due to alterations or delays in embryogenesis or placentation with potential relevance for the screening test performance.

O21. First trimester serum placental growth factor and hyperglycosylated human chorionic gonadotropin are associated with later pre-eclampsia

Introduction First trimester serum placental growth factor (PlGF) predicts early-onset pre-eclampsia (diagnosis Objectives To study the accuracy of using combined maternal serum %hCG-h, PlGF, PAPP-A and maternal risk factors in prediction of pre-eclampsia. Methods We determined gestational-age-adjusted concentrations of PlGF, PAPP-A, hCG (Perkin Elmer Wallac Oy, Turku, Finland) and hCG-h (in-house immunofluorometric assay) in maternal serum at 8–13 weeks of gestation. The study consisted of 98 women who later developed pre-eclampsia and 177 pregnant controls. Of the cases, 24 developed preterm pre-eclampsia (diagnosis 160/110mmHg, proteinuria>5g/24h and/or hemolysis, elevated liver enzymes and low platelet count –syndrome). Results Serum PlGF, %hCG-h and PAPP-A were lower in women with later preterm or severe pre-eclampsia than in controls. In receiver-operating characteristics (ROC) curve analysis the area under the curve (AUC) was 0.680 for PlGF, 0.699 for %hCG-h and 0.714 for PAPP-A to predict preterm pre-eclampsia. AUC values for prediction of severe pre-eclampsia were 0.677, 0.702 and 0.677, respectively. When PlGF, %hCG-h and PAPP-A were combined by logistic regression analysis the AUC value was 0.830 for preterm pre-eclampsia and 0.824 for severe pre-eclampsia. Combination of PlGF, %hCG-h, PAPP-A and maternal risk factors including nulliparity and first-trimester mean arterial pressure (MAP) gave an AUC value of 0.805 for preterm pre-eclampsia and 0.882 for severe pre-eclampsia. When PlGF was removed from the analysis the AUC value was 0.803 for preterm pre-eclampsia and 0.878 for severe pre-eclampsia. When %hCG-h was removed the AUC values were 0.756 and 0.840, respectively. Conclusions Combining %hCG-h with PlGF, PAPP-A and maternal risk factors in first trimester improves prediction of later preterm or severe pre-eclampsia. %hCG-h tended to give better AUC values than PlGF when combined with PAPP-A and maternal risk factors. Thus, in first trimester %hCG-h might be superior to PlGF for prediction of preterm or severe pre-eclampsia and it should be further investigated in risk models of pre-eclampsia.

The utility of circulating LHCGR as a predictor of Down's syndrome in early pregnancy.

BackgroundPrevious studies showed that soluble LHCGR/hCG-sLHCGR concentrations in serum or plasma combined with PAPP-A and free βhCG significantly increased the sensitivity of Down’s syndrome screen at early pregnancy without altering the false positive rate. The goal of the present study was to further examine the role of sLHCGR forms as combinatorial markers and to investigate whether sLHCGR could serve as an independent biomarker for Down’s syndrome in first trimester pregnancy screens.MethodsThe PAPP-A, free βhCG, and hCG-sLHCGR concentrations together with nuchal translucency (NT) were measured in 40 Down’s and 300 control pregnancies. The sLHCGR concentration was analysed in 40 Down’s and 206 control pregnancies.ResultsThe hCG-LHCGR in combination with PAPP-A and free βhCG increased the detection rate (DR) by 35% without altering the false positive rate (FPR). The sLHCGR: hCG-sLHCGR ratio alone detected 80% of Down’s pregnancies in first trimester screening, with a false positive rate of 0.5%.ConclusionsWhile measurement of sLHCGR forms in combination with PAPP-A and free βhCG significantly increases the detection rate of Down’s syndrome at first trimester, the ratio of sLHCGR: hCG-sLHCGR acts as an independent marker with a detection rate that is significantly higher than the existing biochemical markers individually for prenatal first trimester screening of Down’s syndrome.

Cys327Cys polymorphism of the PAPP-A gene (pregnancy associated plasma protein A) is related to mortality of long term hemodialysis patients

ObjectivesPAPP-A is an independent mortality predictor of long term hemodialysis patients and a prognostic marker of acute coronary syndrome in general population. Cys327Cys PAPP-A polymorphism (SNP) (rs12375498) was found to be of significance in preeclampsia and the C allele of the PAPP-A C/G SNP (rs13290387) was defined as an independent risk factor for acute myocardial infarction. The aim of the study was to test the role of these PAPP-A SNPs in long term hemodialysis patients. Design and methodsThe studied group consisted of 464 subjects — 319 long term hemodialysis patients (183 men, 136 women, 62±14years) and 145 controls (65 men, 80 women, 49±14years). A subgroup of 211 hemodialysis patients (118 men, 93 women, 63±13years) was prospectively followed up for 4.5years. During the follow up, 111 patients died, 51 of them due to cardiovascular events. PAPP-A SNPs were analyzed by DNA sequencing and serum PAPP-A was measured by TRACE. ResultsBoth SNPs were in Hardy–Weinberg equilibrium. Allelic and genotype frequencies did not differ between patients and controls and were not related to serum PAPP-A concentrations. Cys327Cys SNP was significant for patients' survival (HR (95% CI): 1.616 (1.110–2.353), nominal p=0.012, corrected p=0.036) while C/G SNP was not. ConclusionsOur study shows for the first time the significance of Cys327Cys PAPP-A SNP (rs12375498) for overall mortality of long term hemodialysis patients. Although it does not influence the concentration of PAPP-A it still could affect the correct function of this enzyme which has to be clarified in further studies.

Regression of the inflammatory microenvironment of the peritoneal cavity in women with endometriosis by GnRHa treatment

ObjectiveTo investigate the effect of gonadotropin-releasing hormone analogues (GnRHa) on the peritoneal fluid microenvironment in women with endometriosis. Study designPeritoneal fluid was collected from 85 women with severe endometriosis (rAFS stage III and IV) during laparoscopic surgery during the proliferative phase. Prior to surgery clinical data were collected. The concentrations of specific markers for endometriosis in the peritoneal fluid were determined using an ELISA and a comparison between peritoneal fluid markers in women using GnRHa and no hormonal treatment was performed using a non-parametric Mann–Whitney U test. ResultsThe study included peritoneal fluid from 39 patients who had been administered GnRHa (Zoladex®) in the three months prior to surgery and 46 from women with no hormonal treatment in this period. Concentrations of IL-8, PAPP-A, glycodelin-A and midkine were significantly reduced in the GnRHa treatment group compared to women receiving no hormonal treatment. RANTES, MCP-1, ENA-78, TNF-α, OPG, IP-10 and defensin showed no significant change between the two groups. ConclusionsGnRHa mediate a significant regression in the inflammatory nature of the peritoneal microenvironment in women with endometriosis.

The association between gestational diabetes mellitus and first trimester aneuploidy screening markers

To establish whether maternal serum first trimester concentrations of PAPP-A and free hCGβ are altered in pregnancies that subsequently are diagnosed by an oral glucose tolerance test (OGTT) with gestational diabetes mellitus (GDM). Over the period 2009 and 2011, the results for women who had first trimester screening for aneuploidy were matched with those having an oral glucose tolerance test at 22-26 weeks for suspected GDM. Free hCGβ, PAPP-A and NT MoMs were compared amongst the group having an OGTT with confirmed GDM and those in which GDM was not confirmed. A second comparison group consisted of all non-aneuploidy singleton pregnancies in which no OGTT was performed. During the three-year period, 27,660 singleton pregnancies were screened of which 7429 cases had an OGTT of which 870 cases were classed as GDM by WHO criteria. There was a significant 7-9% reduction in both PAPP-A and free hCGβ MoM in the GDM group compared with either the OGTT non-GDM group or the remaining pregnancies with no known risk factors for evidence of GDM. There was no difference in the NT measurements. First trimester concentrations of PAPP-A and free hCGβ are reduced in pregnancies that subsequently are diagnosed with GDM and may be useful in further screening algorithms for this disorder although the sensitivity alone is quite poor.

Additive effect of factors related to assisted conception on the reduction of maternal serum pregnancy-associated plasma protein A concentrations and the increased false-positive rates in first-trimester Down syndrome screening

To analyze whether assisted conceptions need adjustments in first-trimester Down syndrome screening and why modifications in screening markers occur. Eleven-year cohort retrospective analysis. Maternal-fetal medicine unit. Two thousand eleven naturally conceived normal singleton pregnancies and 2,042 normal singleton pregnancies achieved with assisted conception: 350 by IUI and 1,692 with IVF (n = 328) or intracytoplasmic sperm injection (ICSI; n = 1,364), using nondonor (n = 1,086) or donated ova (n = 606), with fresh (n = 1,432) or frozen (n = 260) embryos. Comparison of ultrasound and biochemical markers of first-trimester Down syndrome screening according to the mode of conception and considering the clinical and laboratory parameters related. Nuchal translucency (NT), PAPP-A and free βhCG maternal serum concentrations, and false-positive rates (FPRs). NT is unaffected by the mode of conception. Singleton pregnancies achieved by IVF and ICSI with nondonor oocytes have reduced maternal serum PAPP-A and increased FPR, which are significant only in ICSI cycles. Pregnancies from frozen embryos with hormone therapy also show decreased PAPP-A but without affecting the FPR. Elevated maternal serum fβhCG levels in oocyte donation do not influence the FPR. Among assisted conceptions, only nondonor IVF/ICSI singleton pregnancies need adjustments of the maternal serum PAPP-A in first-trimester Down syndrome screening.

PM.10 Are Low PAPP-A Concentrations Associated with Placentally-Mediated Complications in Women with Antiphospholipid Syndrome (APS)?

Women with antiphospholipid syndrome (APS) have pregnancies complicated by placentally-mediated events whereas women with antiphospholipid antibodies (aPL) have similar pregnancy outcomes to the normal population. Low PAPP-A concentrations are useful...

Second trimester amniotic fluid adiponectin level is affected by maternal tobacco exposure, insulin, and PAPP-A level

Adiponectin is an adipocyte-derived plasma protein with insulin-sensitizing and antiatherosclerotic properties. The objectives of the present study were to determine the amniotic fluid (AF) concentration of adiponectin during the second trimester of pregnancy and to demonstrate its association with maternal and fetal variables and AF concentrations of insulin, leptin, and pregnancy-associated-plasma-protein A (PAPP-A). We performed a cross-sectional study of 222 pregnant women who underwent amniocentesis at 15-18 weeks for genetic reasons. No malformation or chromosomal disorder was found in the newborn after birth. AF adiponectin, leptin, PAPP-A, and insulin concentrations were measured using commercially available assays. All maternal, fetal, and biochemical variables were studied using univariate and multivariate linear regression analysis to determine their association with the AF concentration of adiponectin. Adiponectin concentration was negatively correlated with maternal smoking status (β=-5.208; p<0.001) and positively correlated with levels of insulin (β=0.621; p=0.002) and PAPP-A (β=40.150; p<0.001). Non-significant correlations were found between adiponectin concentration and maternal age, maternal body mass index, gestational age at amniocentesis, fetal gender, and AF level of leptin. These findings suggest that the fetus and its membrane adipocytokines, in relationship with maternal and other fetal variables, play a dynamic role in the regulation of energy and oxidative stress homeostasis due to its insulin-sensitizing and antiatherosclerotic effects. The association of these molecules with maternal tobacco consumption during pregnancy could have perinatal implications.

Comparison of Three First Trimester Screening Algorithms for Trisomy 21 with and without Adjustment for Maternal Characteristics

Comparison of three algorithms (DoE 2007 and DoE 2011 algorithm of the FMF Germany and MoM algorithm of the FMF UK) in first trimester biochemical screening for trisomy 21 based on maternal and gestational age, free ß-hCG, and PAPP-A and assessment of relevant maternal characteristics. Data from 22 449 euploid singleton pregnancies undergoing combined screening for trisomy 21 at 11 to 13 weeks of gestation were examined. The measured maternal free β-hCG and PAPP-A concentrations were converted into DoE 2007 and DoE 2011 values according to the algorithm of the FMF Germany and into MoM values according to the algorithm of the FMF UK. In each pregnancy, patient-specific risks and false-positive rates (FPR) were computed according to the three algorithms and were stratified according to gestational age, maternal ethnicity, maternal weight, and smoking status. Free ß-hCG and PAPP-A MoM and DoE 2011 were acceptably independent from maternal characteristics and gestational age, while there was a strong relationship between maternal weight and the DoE 2007 values. For a risk cut-off that corresponds to an overall 5 % FPR rate for each algorithm, the FPR in each group were around 5 % at gestational week 11 - 13. The FPR of the DoE 2007 algorithm increased linearly with maternal weight from 3.6 % in women of 50 kg or less to 11.8 % in women of more than 110 kg. Especially maternal weight has a significant impact on the risk calculation. In contrast to the DoE 2007 algorithm, the DoE 2011 and MoM algorithms both adjust for maternal weight.

Maternal serum human chorionic gonadotropin and pregnancy‐associated plasma protein‐A in pregnancies with fetal homozygous α‐thalassemia‐1 disease

To compare the levels of maternal serum free β-hCG and PAPP-A between pregnancies with fetal homozygous α-thalassemia-1 disease and unaffected pregnancies. One hundred and forty eight pregnancies at risk for fetal homozygous α-thalassemia-1 disease scheduled for cordocentesis at 18 to 22 weeks were recruited into the study. Maternal serum free β-hCG and PAPP-A concentrations were measured before cordocentesis, and the final fetal diagnosis of homozygous α-thalassemia-1 disease was based on fetal Hb typing using high-performance liquid chromatography. Maternal serum concentration of free β-hCG was significantly higher in women with fetal homozygous α-thalassemia-1 disease than those with unaffected fetuses (P = 0.018), whereas the concentrations of PAPP-A was not significantly different (P = 0.184). The median MoM of free β-hCG in the affected group was 1.38 MoM and in the unaffected group was 0.88 MoM (P = 0.020). At midpregnancy, maternal serum free β-hCG levels are significantly higher in pregnancies with fetal homozygous α-thalassemia-1 disease, signifying that the disease could be a confounder for interpretation free β-hCG level in Down syndrome screening program. Nevertheless PAPP-A level seems to be similar in both groups.

Effect of Mild Hepatic or Renal Impairment on Maternal Serum Screening Biochemical Measures

BackgroundIntegrated maternal serum screening (MSS) is commonly used to screen for fetal trisomies and neural tube defects in early pregnancy. The kidney and liver each play an important role in hormone metabolism, and anecdotal data suggest that MSS biochemical measures may vary with a mother’s health status. We examined the correlations between kidney and liver function parameters and MSS markers and the possible association of mild renal or hepatic impairment with MSS measures. MethodsWe completed a prospective cross-sectional study of 257 consecutive women who underwent integrated MSS at a single hospital Serum analytes (pregnancy associated plasma protein A [PAPP-A], hCG, creatinine [Cr], and alanine aminotransferase [ALT]) were drawn at approximately 12 weeks’ gestation, and alpha-fetoprotein and unconjugated estriol were drawn at 16 weeks’ gestation Creatinine clearance was calculated using the Cockcroft-Gault formula Abnormally elevated serum Cr and ALT were each defined as ≥90th percentile among all women. A low creatinine clearance (CrCl) was set at ≤10th percentile. ResultsSerum hCG, PAPP-A, and alpha-fetoprotein were negatively correlated with CrCl, but not after correction for maternal age, weight, and ethnicity. No association between MSS and serum ALT was observed The median serum concentrations of both PAPP-A (P=0.04) and alpha-fetoprotein (P=0.02) were significantly higher among those whose CrCl was≤10th percentile. At the more extreme concentrations of PAPP-A and alpha-fetoprotein, no significant association with a low CrCl or an elevated serum ALT was seen. ConclusionsAmong a group of apparently healthy pregnant women, mild renal or hepatic impairment had little or no significant correlation with individual MSS markers. Further work should focus on the effect of more severe renal or hepatic dysfunction on MSS measures.

The secretion of PAPP-A, ADAM12, and PP13 correlates with the size of the placenta for the first month of pregnancy

ObjectivesPregnancy Associated Protein A (PAPP-A), A Disintegrin and Metalloproteinase 12 (ADAM12) and Placental Protein 13 (PP13) are secreted from the placental trophoblastic tissue and are involved in normal implantation and placental development. The aim of the study was to assess the connection between the secretion of these proteins and the growth of the gestational sac and the placenta. Study designIn an observational longitudinal study at Oulu University Hospital, women with naturally conceived pregnancies were followed-up weekly to pregnancy week 11. Main outcome measuresPAPP-A, ADAM12 and PP13 serum concentrations and their correlation with the volumes of the gestational sac and the placenta were assessed using three-dimensional ultrasonography. ResultsThe study group consisted of 41 women. The PAPP-A, ADAM12 and PP13 serum concentrations increased continuously from pregnancy week 4 to week 11 and correlated closely with each other. The serum concentrations of PAPP-A, ADAM12 and PP13 also correlated with the volumes of the gestational sac and the placenta up to pregnancy week 8. ConclusionsThe secretion of PAPP-A, ADAM12 and PP13 is closely related to the size of the placenta in the beginning of pregnancy. After 8 weeks of pregnancy, which is the time for luteoplacental shift, the correlation disappears, possibly reflecting the morphologic transformation in the placenta.

First trimester serum markers stability during sample transportation from the obstetrical site to the screening laboratory

Objective: To investigate the effect of temperature on first trimester free β-hCG and PAPP-A in serum during transportation. Methods: Maternal blood was collected from 30 East Asian women attending for Down syndrome screening at the primary obstetrical site. The extracted serum was split into two equal aliquots, placed in identical thermally insulated pouches with a temperature data logger and shipped to the screening laboratory. An 800 g frozen ice pack was placed in one of the pouches. Free β-hCG and PAPP-A levels were determined in the ‘Cooled’ and ‘Uncooled’ serum samples using a DELFIA® Xpress analyzer. The proportion of ‘Uncooled’ samples not within 5% of the ‘Cooled’ sample marker level was assessed. Results: The median age of subjects was 36 years, half were nulliparous and all were non-smokers. The assay stability ranged from 2.38% to 4.41%. The median total transport time from the obstetrical site to the laboratory was 17.37 h. Median recorded temperatures within the ‘Cooled’ pouch were significantly lower throughout the transportation of the samples (p < 0.0001). The median percentage change of free β-hCG and PAPP-A concentrations in ‘Uncooled’ samples relative to ‘Cooled’ samples were 2.72% (range 0.27–7.64%) and 1.10% (range 0.03%–4.29%), respectively. Free β-hCG concentrations differed by greater than 5% in eleven (37%) subjects. Conclusions: Insulated packaging, cooling media and temperature monitoring devices may be needed as additional measures to determine, minimize and exclude effects of ambient temperature on serum marker levels especially when travelling times are long or ambient temperatures are high.

Pro-Fibrotic Role of the A2B Adenosine Receptor in Human Cardiac Fibroblasts

Background and Aims: Adenosine is a signaling nucleoside and the interstitial levels of this nucleoside are elevated in certain pathophysiological conditions. Results of our previous studies suggest that adenosine play an important role in pulmonary fibrosis. The goal of the current study is to determine the potential role of adenosine and its receptor subtypes in cardiac fibrosis using the primary human cardiac fibroblasts (HCF) as a model system. Methods: Expression of adenosine receptors (AdoR), a-smooth muscle actin and a-1 pro-collagen was determined using real-time RT-PCR. The concentrations of IL-6, soluble ST-2 and PAPPA in the cell supernatants were measured using ELISA. The concentration of soluble collagen was determined using Sircol collagen assay. Results: Among the four subtypes of AdoRs, the A2B AdoR was expressed at the highest level in HCF. NECA, a stable analog of adenosine, significantly increased the release of IL-6 in a concentration-dependent manner, with a maximal increase of 2.4 6 0.1 fold over the basal level. In addition, NECA (10mM) increased the expression of a-smooth muscle actin and a-1 pro-collagen, and the production of collagen (1.8 6 0.1 fold induction, from 3.4 6 0.2 to 6.0 6 0.4 mg/ml, p!0.05) from HCF. Furthermore, NECA increased the release of two novel biomarkers of cardiovascular diseases (CVD), soluble ST-2 (1.7 6 0.1 fold induction, from 1.5 6 0.1 to 2.6 6 0.1 ng/ml, p!0.05) and PAPPA (4.46 0.6 fold induction, from 1.46 0.5 to 6.26 0.8 ng/ml, p!0.05). The effects of NECA on release of IL-6, collagen, ST-2, and PAPPA and expression of a-smooth muscle actin and a-1 pro-collagen were completely abolished by a selective A2B AdoR antagonist, GS-6201. Conclusions: This study indicates that the A2B AdoR is the predominant subtype of AdoRs expressed in primary human HCF. Activation of this receptor increases the release of IL-6, production of collagen, expression of fibrotic markers and release of biomarkers of CVD. These findings suggest that A2B AdoR might mediate the fibrotic response in heart diseases.

Effects of heparin and low molecular weight heparin on serum level of pregnancy associated plasma protein A in patients with cerebrovascular disease

Objecfive To investigate the effects of treatment for cerebrovascular disorder patients with heparin and low molecular weight heparin(LMWH) on serum PAPP-A concentrations and provide the basis for evaluating the clinical significance of PAPP-A in the following study.Methods Forty cases with cerebrovascular disease from Qilu Hospital from November 2009 to May 2010 were collected in this study.Blood samples were taken before and after drug administration.All cases were divided into four groups according to situation of medication.Group A consisted of 10 patients who received subcutaneous LMWH anticoagulation therapy, and blood samples were collected before LMWH injection, three hours after subcutaneous LMWH anticoagulation therapy in the first day, the second day and the seventh day and 24 hours after the last injection. Group B consisted of 10 patients who did not receive LMWH therapy, and blood samples were collected immediately after admission, the first day, the second day and the seventh day after admission. Group C consisted of 10 patients with percutaneous carotid intervention who received intravenous heparin at the beginning of stenting, and blood samples were collected from the arterial sheath just before angiography and heparin administration, and at 3, 5, 15, 40 and 100 min after heparin administration. Group D consisted of 10 patients who received carotid angiography but LMWH-free therapy,and blood samples were collected from the arterial sheath just before and after angiography. Serum PAPP-A concentrations were analyzed by ELISA to evaluate the differences of intra-groups and differences at different time points of inter-groups. Results In group A, PAPP-A concentrations were time dependent and elevated gradually from 12. 36 (9. 90-14. 32) mIU/L before LMWH injection to 21.80 (23.50-19.73) mIU/L at the seventh day after injection (M=38. 72, P < 0.01 ). In group C, there was a rapid increase of PAPP-A concentration from 12. 86 ( 9. 67-14. 05 ) mIU/L to 51.56 ( 44. 20-66. 00 ) mIU/L within 5 min after intravenous heparin injection (M=46. 06, P <0. 01 ). The PAPP-A concentration of one week after LMWH administration in group A was 21.80 (23.50-19.73) mIU/L, significantly higher than that in group B [11.81 (9. 21-12. 89) mIU/L] (U<0. O01, P<0.01). The PAPP-A concentration at 15 min after heparin administration in group C was 43.70 (37.70-54. 30) mIU/L, significantly higher than that after angiography in group D [14. 18 (11.25-15. 86) mIU/L] ( U<0. 001, P <0. 01 ). The peak level of blood PAPP-A after subcutaneous LMWH injection was significantly lower than that after intravenous heparin injection. The concentrations in group A and C were 21.80 ( 23.50-19. 73 ) and 51.56 (44. 20-66. 00) mIU/L respectively, and had a significant difference ( U=0. 999, P < 0. 01 ) . Conclusions Both intravenous heparin and subcutaneous LMWH administration induce an increase in serum PAPP-A concentration. The effect of drug should be considered when PAPP-A is selected as an evaluation indicator. Key words: Heparin; Heparin,low-molecular-weight; Pregnancy-associated plasma protein-A; Cerebrovascular disorders

Individualized Correction for Maternal Weight in Calculating the Risk of Chromosomal Abnormalities with First-Trimester Screening Data

In the algorithm developed by the Fetal Medicine Foundation (FMF) Germany designed to evaluate the findings of routine first-trimester screening, the false-positive rate (FPR) was determined for the entire study group without stratification by maternal weight. Based on the data received from the continuous audit we were able to identify an increase in the FPR for the weight-related subgroups of patients, particularly for patients with extremely high body weights. The aim of this study was to demonstrate that the variability of the FPR can be reduced through adjusting the concentrations of free β-HCG and PAPP-A measured in the maternal serum by means of a nonlinear regression function modeling the dependence of these values on maternal weight. The database used to establish a version of the algorithm enabling control of the FPR over the whole range of maternal weight consisted of n = 123 546 pregnancies resulting in the birth of a child without chromosomal anomalies. The group with positive outcomes covered n = 500 cases of trisomy 21 and n = 159 trisomies 13 or 18. The dependency of the serum parameters free β-HCG and PAPP-A on maternal weight was analyzed in the sample of negative outcomes by means of nonlinear regression. The fitted regression curve was of exponential form with negative slope. Using this model, all individual measurements were corrected through multiplication with a factor obtained as the ratio of the concentration level predicted by the model to belong to the average maternal body weight of 68.2 kg, over the ordinate of that point on the regression curve which belongs to the weight actually measured. Subsequently, the totality of all values of free β-HCG and PAPP-A corrected for deviation from average weight were used as input data for carrying out the construction of diagnostic discrimination rules described in our recent paper for a database to which no corrections for over- or under-weight had been applied. This entailed in particular the construction of new reference bands for the corrected biochemical values as the basis for calculating the degree of extremeness (DOE) measures to replace the more traditional MOMs. In the final and most crucial step, stratified FPRs were computed and compared over a set of intervals partitioning the whole range of maternal weight into 18 classes. For the posterior risks of both trisomy 21 and 13 / 18 computed from the weight-corrected database, the use of a cutoff value of 1:150 turned out to be an appropriate choice. For T 21, the overall FPR obtained through comparing the individual risks with this cutoff was found to be 3.51 %. The corresponding proportion of ascertained cases of trisomy 21 detected by means of the new algorithm was 86.2 %. For the trisomy 13 / 18 group, the analogous results were a FPR of 2.07 % and a detection rate (DTR) of 83.0 %, respectively. A comparison between the FPRs obtained for the 18 intervals into which the range of maternal weight had been partitioned, showed the deviation of the strata-specific from the overall FPR to be fairly small: for T 21, the FPR ranged from 2.72 to 4.86 %, and the maximum was found in the group of 87.5 - 95.0 kg. For women with a weight of more than 120 kg, the FPR was only slightly above the FPR for the total sample (3.69 as compared to 3.51 %). Similar results were obtained for the discrimination rule constructed for diagnosing T 13 / 18: here, the minimum FPR (1.17 %) was found for patients weighing more than 120 kg, whereas the maximum (2.66 %) occurred in the interval 75.0 - 77.5 kg. In this study we demonstrated that the new algorithm developed by the FMF Germany to estimate risks for fetal trisomies 21 and 13 / 18 combines very good misclassification rates with a far-reaching stability of the false-positive rate against even extreme deviations from the average maternal weight.

The impact of temporal variability of biochemical markers PAPP-A and free β-hCG on the specificity of the first-trimester Down syndrome screening: a Croatian retrospective study

BackgroundThe variability of maternal serum biochemical markers for Down syndrome, free β-hCG and PAPP-A can have a different impact on false-positive rates between the 10+0 and 13+6 week of gestation. The study population comprised 2883 unaffected, singleton, spontaneously conceived pregnancies in Croatian women, who delivered apparently healthy child at term. Women were separated in 4 groups, dependently on the gestational week when the analyses of biochemical markers were performed. The concentrations of free β-hCG and PAPP-A in maternal serum were determined by solid-phase, enzyme-labeled chemiluminiscent immunometric assay (Siemens Immulite). Concentrations were converted to MoMs, according to centre-specific weighted regression median curves for both markers in unaffected pregnancies. The individual risks for trisomies 21, 18 and 13 were computed by Prisca 4.0 software.FindingsThere were no significant differences between the sub-groups, regarding maternal age, maternal weight and the proportion of smokers. The difference in log10 MoM free β-hCG values, between the 11th and 12th gestational week, was significant (p = 0.002). The difference in log10 MoM PAPP-A values between the 11th and 12th, and between 12th and 13th week of gestation was significant (p = 0.006 and p = 0.003, respectively). False-positive rates of biochemical risk for trisomies were 16.1% before the 11th week, 12.8% in week 12th, 11.9% in week 13th and 9.9% after week 13th. The differences were not statistically significant.ConclusionsBiochemical markers (log10 MoMs) showed gestation related variations in the first-trimester unaffected pregnancies, although the variations could not be attributed either to the inaccuracy of analytical procedures or to the inappropriately settled curves of median values for the first-trimester biochemical markers.