The relationship between the levels of gonadotropic hormones and OPG,leptin, TGF-β1 and TGF-β2 in Chinese adult women

BackgroundThe relationship between the levels of gonadotropic hormones and bone metabolism-related cytokines in Chinese women is unclear. We investigated the relationship between FSH and LH and OPG, leptin, TGF-β1, and TGF-β2 in Chinese women.MethodsA cross-sectional study of 694 Chinese women, aged 20 to 82 y was conducted. Levels of serum FSH, LH, OPG, leptin, TGF-β1, and TGF-β2 were determined.ResultsIn premenopausal females, serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels seemly showed no correlation with the cytokine levels. In perimenopausal females, serum FSH and LH levels showed significant positive correlation with osteoprotegerin (OPG) and transforming growth factor-β2 (TGF-β2) levels (r = 0.286 to 0.405, all P = 0.000), whereas they showed negative correlation with TGF-β1 levels (r = ? 0.413 and ? 0.354, all P = 0.000). In postmenopausal females, FSH and LH levels showed positive correlation with OPG levels (r = 0.247 and 0.241, all P = 0.000), negative correlation with leptin and TGF-β1 levels (r = ? 0.234 to ? 0.319, all P = 0.000), and no correlation with TGF-β2 levels. Multiple linear regression stepwise analysis revealed the following results. In premenopausal females, 2.0% and 1.5% of the changes in LH could be explained by OPG and leptin, respectively, while 1.9% of the changes in OPG could be explained by LH. In perimenopausal females, the determinants of OPG and TGF-β1 on FSH were 10.9% and 17.0%, respectively, and the determinants of OPG, TGF-β1 and TGF-β2 on LH were 4.5%, 4.9% and 16.4%, respectively. The determinants of FSH and LH on OPG were 14.5% and 2.5%, respectively. The determinant of FSH on TGF-β1 was 4.5%, while the determinant of LH on TGF-β2 was 16.4%. In postmenopausal females, the determinants of leptin and OPG on FSH were 10.2% and 2.8%, respectively, and the determinants of OPG and TGF-β1 on LH were 5.8% and 2.3%, respectively. The determinant of FSH on OPG, leptin and TGF-β1 were 6.1%, 3.4% and 9.2%.ConclusionsThese results indicate that age-related gonadotropic hormone levels are associated with changes in OPG, TGF-β1, TGF-β2 and leptin, and change with menopausal status.     

The relationship between bone turnover markers and BMD decreasing rates in Chinese middle-aged women

BackgroundThe relationship between bone turnover markers (BTMs) and BMD decreasing rate (BDR) in Chinese women is unclear. Wu investigated the relationship between (BTMs) and BDR at various skeletal sites in Chinese middle-aged women.MethodsA cross-section study of 555 healthy Chinese women over 35–60 years of age. BMD at posteroanterior spine, the left hip, and the left forearm were measured with a DXA. Levels of serum osteocalcin (OC), bone-specific alkaline phosphatase (BAP), cross-linked N-terminal telopeptides of type I collagen (sNTX) and total urinary deoxypyridinoline (uDPD) were determined.ResultsBDR at various skeletal sites had significant negative correlation with serum OC(r = ? 0.395 to ? 0.530), BAP(r = ? 0.297 to ? 0.486), and sNTX(r = ? 0.207 to ? 0.272). After adjustment of age and weight, serum OC, BAP, and sNTX rather than total uDPD still exhibited significant correlations with BDR. Stepwise regression analyses showed that, serum OC and BAP were the significantly negative determinants of BDR. Between 4.7?27.7% and 1.2?16.1% of the changes in BDR were determined by serum OC and BAP, respectively. However, sNTX and total uDPD had no significant effect on BDR at various skeletal sites.ConclusionsThis study indicated the correlation between BTMs and early-stage BDR in Chinese middle-aged women and suggested that serum OC and BAP, rather than sNTX and total uDPD, are the key determining factors of early BMD decreases.     

Glycated albumin is independently associated with estimated glomerular filtration rate in nondiabetic patients with chronic kidney disease

BackgroundGlycated albumin (GA) may contribute to diabetic nephropathy, but the clinical significance of GA in patients with chronic kidney disease (CKD) is unknown.MethodsPatients were classified with the NKF/DOQI classification system as mild (stage I, II), moderate (stage III), or advanced CKD (stage IV). Those undergoing dialysis or with CKD stage V were excluded. GA was measured using the Lucica TM GA-L assay kit. The relationship between GA and renal dysfunction was analyzed in patients with or without diabetes.ResultsA total of 187 subjects were enrolled. GA values in those with normal, mild, moderate and advanced CKD were 18.4 ± 1.4%, 18.4 ± 3.1%, 19.0 ± 3.8%, 20.4 ± 6.4%, respectively, in diabetic patients (N = 67, p = 0.5), and were 14.1 ± 1.9%, 14.2 ± 2.2%, 15.9 ± 1.9%, 15.0 ± 1.7%, respectively, in nondiabetic patients (N = 120, p = 0.004). GA value was negatively correlated to eGFR in nondiabetic patients (r = ?0.35, p < 0.001) but not in diabetic patients (r = ?0.11, p = 0.39). In the adjusted model, GA is independently correlated to eGFR only in nondiabetic subjects.ConclusionsIncreased GA concentrations are independently associated with renal dysfunction in nondiabetic patients with CKD.     

Effect of somatostatin analogue octreotide on pain relief after major abdominal surgery

BackgroundOctreotide acetate is an 8-amino-acids synthetic octapeptide analogue of somatostatin with much-enhanced duration of action and lower incidence of side effects. We assessed the utility of using intravenous octreotide as an adjuvant to opioid analgesia that might exert a post-operative opioid-sparing effect.MethodsForty-four patients were randomly allocated, to receive either a placebo or intraoperative octreotide 0.33 μg kg^?1 h^?1 intravenous infusion that was maintained in the post-operative period. Patients received for post-operative analgesia an intravenous piritramide patient controlled analgesia (PCA), set to deliver a piritramide 0.02 mg kg^?1 dose.ResultsTwo-way ANOVA revealed significantly fewer (P = 0.0003) mean ± SD weighted piritramide dose requirements in the octreotide group (19.5 ± 6.3 μg kg^?1 h^?1) than in the control group (35.7 ± 8.2 μg kg^?1 h^?1). Dunnett’s two-sided multiple-comparison post hoc test revealed a significant difference between the two groups during the first 22 post-operative hours, following which there were no differences between the two groups. There were no significant differences over time in the mean arterial pressure (P = 0.722), heart rate (P = 0.579) and respiratory rate (P = 0.823) between the octreotide group (80 ± 10 mm Hg, 74 ± 12, 14 ± 2) and the control group (82 ± 9 mm Hg, 76 ± 11, 15 ± 3), respectively.ConclusionWe demonstrated that perioperative octreotide intravenous infusion could be an adjuvant to opioid analgesia as it exerted a piritramide opioid-sparing effect. We encountered more systemic side effects such as nausea, abdominal discomfort, and diarrhea in the octreotide group than in the control group. Our findings could be beneficial to patients who cannot tolerate the adverse effects of opioids.     

Comparison of normal saline, hypertonic saline and hypertonic saline colloid resuscitation fluids in an infant animal model of hypovolemic shock

PurposeIncorrect resuscitation after hypovolemic shock is a major contributor to preventable pediatric death. Several studies have demonstrated that small volumes of hypertonic or hypertonic–hyperoncotic saline can be an effective initial resuscitation solution. However, there are no pediatric studies to recommend their use. The aim of this study is to determine if in an infant animal model of hemorrhagic shock, the use of hypertonic fluids, as opposed to isotonic crystalloids, would improve global hemodynamic and perfusion parameters.MethodsExperimental, randomized animal study including thirty-four 2-to-3-month-old piglets. 30 min after controlled 30 mL kg^?1 bleed, pigs were randomized to receive either normal saline (NS) 30 mL kg^?1 (n = 11), 3% hypertonic saline (HS) 15 mL kg^?1 (n = 12), or 5% albumin plus 3% hypertonic saline (AHS) 15 mL kg^?1 (n = 11).ResultsHigh baseline heart rate (HR) and low mean arterial pressure (MAP), cardiac index (CI), brain tissue oxygenation index (bTOI), and lactate were recorded 30 min after volume withdrawal, with no significant differences between groups. Thirty minutes after volume replacement there were no significant differences between groups for HR (NS, 188 ± 14; HS, 184 ± 14; AHS, 151 ± 14 bpm); MAP (NS, 80 ± 7; HS, 86 ± 7; AHS, 87 ± 7 mmHg); CI (NS, 4.1 ± 0.4; HS, 3.9 ± 0.4; AHS, 5.1 ± 0.4 mL min^?1 m^?2); lactate (NS, 2.8 ± 0.7; HS, 2.3 ± 0.6; AHS, 2.4 ± 0.6 mmol L^?1); bTOI (NS, 43.9 ± 2.2; HS, 40.1 ± 2.5; AHS, 46.1 ± 2.3%).ConclusionsIn this model of hypovolemic shock, hypertonic fluids achieved similar end-points as twice the volume of NS. Animals treated with albumin plus hypertonic saline presented prolonged increase in blood volume parameters and recovery of the oxygen debt.     

Gamma-glutamyltransferase, alanine transaminase and aspartate transaminase levels and the diagnosis of gestational diabetes mellitus

ObjectivesTo evaluate gamma-glutamyltransferase (GGT), alanine transaminases (ALT) and aspartate transaminases (AST) levels and prevalent gestational diabetes mellitus (GDM).Design and methodsRandom plasma glucose, GGT, ALT and AST and the 50-g glucose challenge test were done on antenatal women followed by diagnostic 3-point 75-g oral glucose tolerance test within two weeks. GDM was diagnosed by ADA (2011) criteria.ResultsThe GDM rate was 12.2% (319/2610). Mean GGT level was higher in GDM women, 18 ± 12 vs. 16 ± 11 IU/L; P = 0.03. The risk for GDM was higher for women in the highest GGT quartile band compared to the lowest: RR 1.35 95%CI 1.0–1.8; P = 0.04. However, after adjustment for confounders, GGT was no longer associated with GDM. There was no correlation between ALT and AST levels and GDM.ConclusionsLiver transaminases do not predict GDM in contrast to type 2 diabetes.     

Use of independent component analysis for reducing CPR artefacts in human emergency ECGs

AimWe applied independent component analysis (ICA) to cardiopulmonary resuscitation (CPR)-corrupted human multichannel emergency ECGs with the aim of reconstructing the original ECGs.MaterialsTwo ICA algorithms (EFICA and JADE) were selected. Data for ICA were acquired by simultaneously recording eight ECG channels during CPR on a porcine model. The algorithms’ reconstruction performance was assessed by the Spearman correlation coefficient (SCC) and the shock advice algorithm of an AED. We then compared the performance of EFICA with the established second-channel adaptive matching pursuit method (AF).ResultsICA was applied to 918 corrupted ECG multichannel signals. The sensitivity of the AED's shock/no-shock decision increased from 93.5% (corrupted signal) to 99.5/99.8% (JADE/EFICA) in the selected independent component; specificity increased from 50.5% to 78.9/83.2% (JADE/EFICA). The SCCs comparing the reconstructed with the original signal (JADE: 0.75 ± 0.15; EFICA: 0.76 ± 0.15, n = 918) were significantly higher than for the corrupted signal vs. the original (0.52 ± 0.22). The SCC is significantly higher (p < 0.01) using EFICA than AF (EFICA: 0.75 ± 0.16; AF: 0.72 ± 0.19, n = 718). For all signals at all SNR levels, specificity did not differ significantly between EFICA (83.6%) and AF (80.2%). EFICA proved to be superior especially at low corruption levels (SNR < ?5 dB). Sensitivity was above 99.5% for both algorithms.ConclusionWe have demonstrated that CPR artefacts in the emergency ECG can be reduced using ICA. EFICA and JADE are at least as successful in this regard as are other published algorithms. In particular, non-shockable signals with low SNRs ( dB) are reconstructed significantly better (p = 0.01) with EFICA than with AF.     

Decreased serum brain-derived neurotrophic factor concentration in young nonobese subjects with low insulin sensitivity

ObjectiveInsulin resistance and type 2 diabetes are associated with an increased risk of neurodegenerative diseases. Decreased brain-derived neurotrophic factor (BDNF) levels might play a role in the pathogenesis of neuropsychiatric disorders. The aim of our study was to estimate serum BDNF concentration in nonobese women divided into subgroups according to their insulin sensitivity.Design and methodsWe studied 46 young, healthy, nonobese women. Insulin sensitivity was estimated with the euglycemic–hyperinsulinemic clamp technique. Then, participants were divided into subgroups of high (mean, 12.79 ± 2.01 mg/kg fat-free mass/min) and low insulin sensitivity (mean, 7.33 ± 1.66 mg/kg fat-free mass/min).ResultsWe observed decreased serum BDNF concentration in women with low insulin sensitivity in comparison to high insulin sensitivity group (3306.11 ± 603.10 vs 4141.91 ± 755.37 pg/mL, p = 0.001). Serum BDNF was positively related to insulin sensitivity (r = 0.43, p = 0.003). This correlation remained significant after adjustment for other estimated parameters.ConclusionsSerum BDNF is decreased in young nonobese women with low insulin sensitivity. Early detection and prevention of insulin resistance might be useful in the prevention of neurodegenerative disorders.     

The circulating concentration and ratio of total and high molecular weight adiponectin in post-menopausal women with and without osteoporosis and its association with body mass index and biochemical markers of bone metabolism

ObjectivesThere is increasing evidence suggesting that adiponectin plays a role in the regulation of bone metabolism.Design and methodsThis was a cross-sectional study of 34 post-menopausal women with and 37 without osteoporosis. All subjects had body mass index (BMI), bone mineral density (BMD), total-, high molecular weight (HMW)-adiponectin and their ratio, osteoprotegerin (OPG), a marker of bone resorption (βCTX) and formation (P1NP) measured.ResultsWe observed a positive correlation between BMI and BMD (r = 0.44, p < 0.001). When normalised for BMI, total-, HMW-adiponectin concentrations and HMW/total-adiponectin ratio were significantly lower in obese compared to lean subjects but there was no difference between those with or without osteoporosis. There were significant negative correlations between HMW/total-adiponectin ratio and BMI (r = ? 0.27, p = 0.030) and with OPG (r = ? 0.44, p < 0.001).ConclusionsOur data suggests that there is no significant difference in the circulating concentration of fasting early morning total- or HMW-adiponectin in post-menopausal women with or without osteoporosis. The correlation between HMW/total-adiponectin ratio and OPG may indicate that adiponectin could influence bone metabolism by altering osteoblast production of OPG thereby affecting osteoclasts mediated bone resorption.     

Diabetes in pregnancy: Effect on glycation and acetylation of the different chains of fetal and maternal hemoglobin

BackgroundMeasurement of glycated fetal hemoglobin to assess maternal glycemic control is unreliable. Electrospray ionization–mass spectrometry (ESI-MS) has been suggested as a definitive procedure.ObjectiveThis study aimed to evaluate glycation and acetylation by ESI-MS of the separate chains of neonatal fetal hemoglobin in comparison with glycation of maternal hemoglobin, to assess the impact of maternal diabetes.MethodsTwenty-nine non-diabetic (31 neonates) and 15 diabetic women (15 neonates) were recruited. Whole blood was collected at delivery from the mothers and cord blood from their respective neonate. The blood samples were diluted in acetonitrile:water (50:50) and treated with a cation exchange resin to remove sodium and potassium adducts on the hemoglobin. The α- and β-chain glycated maternal hemoglobin and α- and γ-chain glycated and acetylated hemoglobin of the neonate were measured by ESI-MS. HbA1c was measured on the Menarini 8160.ResultsMean α- and γ-chain and overall glycated hemoglobin and acetylated fetal hemoglobin were 1.23 ± 0.41%, 1.62 ± 0.47%, 1.24 ± 0.37%, and 8.56 ± 0.84% in the infant of the non-diabetic mother (INDM) and 1.39 ± 0.21%, 1.92 ± 0.61%, 1.64 ± 0.59%, and 8.44 ± 0.63% in the infant of the diabetic mother (IDM). Mean maternal α- and β-chain, overall glycated hemoglobin and HbA1c were 1.98 ± 0.38%, 4.28 ± 0.76%, 3.13 ± 0.51%, 5.39 ± 0.39% (non-diabetic) and 2.40 ± 0.83%, 4.71 ± 0.90%, 3.58 ± 0.83%, 6.15 ± 0.71% (diabetic). Overall glycated hemoglobin levels were significantly higher in the IDM (p = 0.006) compared to the INDM. Maternal glycated hemoglobin was significantly higher than neonatal glycated hemoglobin in the control (p < 0.0001) and diabetic group (p < 0.0001). A significant correlation was observed between maternal glucose concentration and overall glycated fetal hemoglobin in the IDM (p = 0.02). Glucose concentrations in the diabetic mother and the IDM were not significantly different (p = 0.5) and were significantly correlated (p < 0.0001). HbA1c was not significantly different in the two maternal groups. No significant difference was observed between AcHbF in IDM or INDM (p = 0.61).ConclusionsMeasurement of overall glycated fetal hemoglobin, incorporating α- and γ-chain glycations, seems best to assess abnormal glucose homeostasis during pregnancy. Accurate assay of this parameter is critical for stratification of risk of possible post birth developmental complications.     

Carotid intima media thickness is related positively to plasma pre ß-high density lipoproteins in non-diabetic subjects

BackgroundLipid-poor or lipid-free high density lipoprotein (HDL) particles, designated pre ß-HDL, stimulate removal of cell-derived cholesterol to the extracellular compartment, which is an initial step in the reverse cholesterol transport pathway. Pre ß-HDL levels may be elevated in subjects with established cardiovascular disease. We determined the relationship of carotid intima media thickness (IMT), a marker of subclinical atherosclerosis, with pre ß-HDL in subjects without clinically manifest cardiovascular disease.MethodsIMT and plasma pre ß-HDL, assayed by crossed immuno-electrophoresis, were determined in 70 non-diabetic subjects (aged 56 ± 9 years; non-smokers only; 27 women).ResultsIMT was correlated positively with pre ß-HDL, both expressed as plasma apolipoprotein (apo) A-I concentration (r = 0.271, p = 0.023) and as% of apo A-I (r = 0.341, p = 0.004). In contrast, IMT was correlated inversely with HDL cholesterol (r = ? 0.253, p = 0.035). IMT was also related positively to pre ß-HDL after adjustment for age, sex, systolic blood pressure (in apoA-I concentration, ß = 0.203, p = 0.043; in% of plasma apoA-I, ß = 0.235, p = 0.023). IMT remained associated with pre ß-HDL after additional adjustment for either body mass index, plasma glucose, cholesterol, triglycerides, HDL cholesterol, apoA-I and apoB.ConclusionSubclinical atherosclerosis may relate to higher plasma pre ß-HDL independently of apoA-I and HDL cholesterol levels.     

Serum ionic fluoride concentrations are significantly decreased after treatment with alendronate in patients with osteoporosis

BackgroundWe determined serum ionic fluoride (SIF) concentrations before and after treatment of osteoporosis with alendronate to clarify whether SIF concentrations directly reflect a change in bone metabolism.MethodsA total of 45 postmenopausal women with primary osteoporosis who were treated with alendronate over a 6-month period were enrolled (mean age, 64.2 years). SIF concentrations were measured by the flow injection method with an ion-selective electrode. Concentrations of bone turnover markers (serum bone alkaline phosphatase, serum osteocalcin, serum type I collagen cross-linked N-telopeptide and urinary deoxypryridinoline) and lumbar spine BMD (LsBMD) were also measured. SIF, bone turnover markers and LsBMD before and after treatment were compared.ResultsConcentrations of SIF as well as concentrations of all bone turnover markers were significantly decreased after treatment: means (standard deviations) before and after treatment were 0.62 (0.13) and 0.32 (0.09) μmol/l, respectively (P < 0.001) and the percent change was ? 46.3%. LsBMD was also significantly increased by 6.7% after treatment.ConclusionsThe reduction of SIF concentrations is probably caused by inhibition of bone resorption due to the action of alendronate. The findings suggest that SIF concentrations directly reflect a change in bone metabolism.     

Effect of implementation of new resuscitation guidelines on quality of cardiopulmonary resuscitation and survival

BackgroundDuring cardiopulmonary resuscitation (CPR), advanced life support (ALS) providers have been shown to deliver inadequate CPR with long intervals without chest compressions. Several changes made to the 2005 CPR Guidelines were intended to reduce unnecessary interruptions. We have evaluated if quality of CPR performed by the Oslo Emergency Medical System (EMS) improved after implementation of the modified 2005 CPR Guidelines, and if any such improvement would result in increased survival.Materials and methodsRetrospective, observational study of all consecutive adult cardiac arrest patients treated during a 2-year period before (May 2003–April 2005), and after (January 2006–December 2007) implementation of the modified 2005 CPR Guidelines. CPR quality was assessed from continuous electronic recordings from LIFEPACK 12 defibrillators where ventilations and chest compressions were identified from transthoracic impedance changes. Ambulance run sheets, Utstein forms and hospital records were collected and outcome evaluated.ResultsResuscitation was attempted in 435 patients before and 481 patients after implementation of the modified 2005 CPR Guidelines. ECGs usable for CPR quality evaluation were obtained in 64% and 76% of the cases, respectively. Pre-shock pauses decreased from median (interquartile range) 17 s (11, 22) to 5 s (2, 17) (p = 0.000), overall hands-off ratios from 0.23 ± 0.13 to 0.14 ± 0.09 (p = 0.000), compression rates from 120 ± 9 to 115 ± 10 (p = 0.000) and ventilation rates from 12 ± 4 to 10 ± 4 (p = 0.000). Overall survival to hospital discharge was 11% and 13% (p = 0.287), respectively.ConclusionQuality of CPR improved after implementation of the modified 2005 Guidelines with only a weak trend towards improved survival to hospital discharge.     

Serum autotaxin measurements in pregnant women: application for the differentiation of normal pregnancy and pregnancy-induced hypertension

BackgroundThe bioactive lipid lysophosphatidic acid (LPA) exerts multiple effects in the female reproductive system. Serum/plasma LPA is mainly produced by the lysophospholipase D activity of autotaxin (ATX). Previous studies have suggested that ATX has critical roles in cancer, reproduction, and vascular development. In the present study, we evaluated the usefulness of serum ATX measurements in pregnant women.MethodsWe measured the serum ATX antigen levels in 32 normal pregnant women, 15 patients with pregnancy-induced hypertension (PIH), and 7 patients with preterm delivery using a recently developed automated enzyme immunoassay.ResultsThe serum ATX antigen levels in normal pregnant women were significantly higher than those in non-pregnant women (P < 0.001). The serum ATX antigen levels in normal pregnant women were significantly and positively correlated with the gestational week (r = 0.809, P < 0.001). During the third trimester, the serum ATX antigen levels of the patients with PIH (3.299 ± 1.720 mg/l) were significantly lower than those of the normal pregnant women (4.915 ± 2.323 mg/l) (P = 0.04).ConclusionsThe serum ATX antigen level increases with the progression of pregnancy. The serum ATX level may be a serological marker for the prediction of PIH.     

Association of vitamin D receptor gene polymorphisms with BMD and their effect on 1, 25-dihydroxy vitamin D3 levels in pre- and postmenopausal South Indian women from Andhra Pradesh

BackgroundOsteoporosis is a multifactorial disorder with a strong genetic component and vitamin D receptor gene has been suggested as a candidate gene for osteoporosis. Therefore the present study was aimed to investigate the role of the VDR Fok 1 gene polymorphism and its influence on vitamin D3 levels and BMD in pre- and postmenopausal osteoporotic women of Indian ethnicity.Methods427osteoporotic women and 460 age matched controls were included in the study. VDR FOK1 gene polymorphism was assessed by the PCR-RFLP method. Serum vitamin D3 was measured by the HPLC method.ResultsThe frequency of ff genotype and f allele was significantly high in pre- and postmenopausal osteoporotic women in comparison with controls (p < 0.001).In each case individuals with ff genotype had significantly low BMD in comparison with Ff and FF genotypes. No significant association was found between the genotypes and vitamin D3 levels.ConclusionThe VDR Fok 1 gene is associated with low bone mass in all our study subjects. The genotype ff of the VDR Fok 1 gene is associated with osteoporosis. Further ff genotype associated significantly with low bone mass. Therefore the f allele of VDR Fok1 gene is an important risk factor for osteoporosis.     

Effects of a single dose of oral iron on hepcidin concentrations in human urine and serum analyzed by a robust LC-MS/MS method

BackgroundThe measurement of serum hepcidin, a peptide hormone that regulates iron metabolism, is clinically important to the understanding of iron homeostasis in health and disease. To date, the quantification of serum hepcidin levels by conventional immunological detection methods has proven problematic due to challenges in obtaining high quality antibodies which demonstrate good reproducibility. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) has been employed recently for more sensitive quantification of hepcidin; however, this method has high background levels and therefore less than optimal specificity.MethodsIn order to increase the specificity of the mass spectrometry based assay, we developed a robust, ultra-performance liquid-chromatography-tandem mass spectrometry (UPLC-MS/MS) protocol using multiple selected reaction monitoring (mSRM) for quantification of hepcidin levels in urine and serum of human subjects. With this assay, we assessed levels of hepcidin before and for up to 8 h after oral ingestion of ferrous sulfate in ten adult human subjects without known disease.ResultsThe linear response of hepcidin quantitation on each instrument was measured, and the correlation coefficients of these calibrations were r² = 0.9512 ± 0.0202 (n = 5) for urine and r² = 0.9709 ± 0.0291 (n = 5) for serum [r² = mean ± SD]. Compared to baseline, the levels of urinary hepcidin between 2–4 h and 4–8 h of both women and men showed significant increases with p < 0.05 and p < 0.001, respectively. The levels of serum hepcidin between 4 h and 8 h in both women and men showed significant increases, compared with baseline values, with both p < 0.01. Interestingly, we also observed some degree of oscillation of levels, occurring at later time points.ConclusionsWe have developed and validated a new method for measuring hepcidin concentrations in human serum and urine and used it to demonstrate early increases with iron supplement in both urinary and serum levels of hepcidin, which return to baseline levels, except in urine samples from men.     

Determination of sensitivity and specificity of a novel gene dosage assay for prenatal screening of trisomy 21 syndrome

ObjectiveTo compare the gene dosage results achieved by a novel multiplex quantitative assay with cytogenetic and quantitative fluorescent polymerase chain reaction (QF-PCR) analysis for prenatal screening of trisomy 21 syndrome on corresponding fetal samples.Design and methodsFetal samples (n = 134) were collected from pregnant women considered high risk for having trisomy 21 affected fetus. Cytogenetic analysis and QF-PCR were performed. Then, the relative gene dosage of DSCAM and DYRK1A2 genes was determined on corresponding samples using comparative delta cycle of threshold (ΔC_T) method.ResultsThe mean gene dosage ratio was 1.55 ± 0.11 (95% CI:1.51–1.58) in trisomy 21 cases and 1.01 ± 0.12 (95% CI:0.98–1.03) in normal samples (p value < 0.001). The results were in agreement to the results of cytogenetic and QF-PCR analysis with the overall specificity of 0.96 (95% CI:0.91–0.98) and the sensitivity of 0.80 (95% CI:0.49–0.94).ConclusionsThis gene dosage assay is appropriate for the screening of high risk pregnant women and is readily amenable to automation.     

Analysis of amplitude spectral area and slope to predict defibrillation in out of hospital cardiac arrest due to ventricular fibrillation (VF) according to VF type: Recurrent versus shock-resistant

BackgroundIn out-of-hospital cardiac arrest (OHCA) due to ventricular fibrillation (VF), VF may recur during resuscitation (recurrent VF) or fail to defibrillate (shock-resistant VF). While retrospective studies have suggested that amplitude spectral area (AMSA) and slope predict defibrillation, it is unknown whether the predictive power is influenced by VF type. We hypothesized that in witnessed OHCA with initial rhythm of VF that the utility for AMSA and slope to predict defibrillation would differ between shock-resistant and recurrent VF.MethodsAMSA and slope were measured immediately prior to each shock. For second or later shocks, VF was classified as recurrent or shock-resistant. Cardiac arrest was classified according to whether the majority of shocks were for recurrent VF or shock-resistant VF.Results44 patients received 98 shocks for recurrent VF and 96 shocks for shock-resistant VF; 24 patients achieved ROSC in the field. AMSA and slope were higher in recurrent VF compared to shock-resistant VF (AMSA: 28.8 ± 13.1 vs 15.2 ± 8.6 mV Hz, P < 0.001, and slope: 2.9 ± 1.4 vs 1.4 ± 1.0 mV s^?1, P = 0.001). Recurrent VF was more likely to defibrillate than shock-resistant VF (P < 0.001). AMSA and slope predicted defibrillation in shock-resistant VF (P < 0.001 for both AMSA and slope) but not in recurrent VF. Recurrent VF predominated in 79% of patients that achieved ROSC compared to 55% that did not (P = 0.10).ConclusionsIn witnessed OHCA with VF as initial rhythm, recurrent VF is associated with higher values of AMSA and slope and is likely to re-defibrillate. However, when VF is shock-resistant, AMSA and slope are highly predictive of defibrillation.     

Atrial natriuretic peptide stability

ObjectiveAtrial natriuretic peptide (ANP) is a key regulator in the homeostasis of water excretion and has emerged as an important prognostic marker for symptomatic chronic heart failure (CHF). The stability of ANP represents a crucial factor in assessing its use as a cardiac biomarker. Accordingly, we assessed the stability of ANP in blood samples collected from healthy controls and CHF subjects for a 12 month period.MethodsBlood samples from 10 healthy controls and 12 symptomatic CHF subjects with left ventricular systolic dysfunction were drawn. Determination of plasma ANP was performed by a standardized radioimmunoassay protocol.ResultsThe ANP levels of healthy subjects were 68.5 ± 11.6 pg/mL at baseline and 69.9 ± 17.2 pg/mL at 12 months (p = 0.71). The ANP concentrations of CHF subjects were 199.25 ± 44.8 pg/mL at baseline and 197.83 ± 47.4 pg/mL at 12 months (p = 0.70) respectively.ConclusionANP is a stable molecule with no evidence of degradation when stored at ? 80 °C.