Genetic markers of repolarization and arrhythmic events after acute coronary syndromes.

BACKGROUND: There is a genetic contribution to the risk of ventricular arrhythmias in survivors of acute coronary syndromes (ACS). We wished to explore the role of 33 candidate single nucleotide polymorphisms (SNPs) in prolonged repolarization and sudden death in patients surviving ACS. METHODS: A total of 2,139 patients (1680 white ethnicity) surviving an admission for ACS were enrolled in the prospective Coronary Disease Cohort Study. Extensive clinical, echocardiographic, and neurohormonal data were collected for 12 months, and clinical events were recorded for a median of 5 years. Each SNP was assessed for association with sudden cardiac death (SCD)/cardiac arrest (CA) and prolonged repolarization at 3 time-points: index admission, 1 month, and 12 months postdischarge. RESULTS: One hundred six SCD/CA events occurred during follow-up (6.3%). Three SNPs from 3 genes (rs17779747 [KCNJ2], rs876188 [C14orf64], rs3864180 [GPC5]) were significantly associated with SCD/CA in multivariable models (after correction for multiple testing); the minor allele of rs17779747 with a decreased risk (hazard ratio [HR] 0.68 per copy of the minor allele, 95% CI 0.50-0.92, P = .012), and rs876188 and rs386418 with an increased risk (HR 1.52 [95% CI 1.10-2.09, P = .011] and HR 1.34 [95% CI 1.04-1.82, P = .023], respectively). At 12 months postdischarge, rs10494366 and rs12143842 (NOS1AP) were significant predictors of prolonged repolarization (HR 1.32 [95% CI 1.04-1.67, P = .022] and HR 1.30 [95% CI 1.01-1.66, P = .038], respectively), but not at earlier time-points. CONCLUSION: Three SNPs were associated with SCD/CA. Repolarization time was associated with variation in the NOS1AP gene. This study demonstrates a possible role for SNPs in risk stratification for arrhythmic events after ACS.

Zebrafish as a model for long QT syndrome: the evidence and the means of manipulating zebrafish gene expression.

Congenital long QT syndrome (LQT) is a group of cardiac disorders associated with the dysfunction of cardiac ion channels. It is characterized by prolongation of the QT-interval, episodes of syncope and even sudden death. Individuals may remain asymptomatic for most of their lives while others present with severe symptoms. This heterogeneity in phenotype makes diagnosis difficult with a greater emphasis on more targeted therapy. As a means of understanding the molecular mechanisms underlying LQT syndrome, evaluating the effect of modifier genes on disease severity as well as to test new therapies, the development of model systems remains an important research tool. Mice have predominantly been the animal model of choice for cardiac arrhythmia research, but there have been varying degrees of success in recapitulating the human symptoms; the mouse cardiac action potential (AP) and surface electrocardiograms exhibit major differences from those of the human heart. Against this background, the zebrafish is an emerging vertebrate disease modelling species that offers advantages in analysing LQT syndrome, not least because its cardiac AP much more closely resembles that of the human. This article highlights the use and potential of this species in LQT syndrome modelling, and as a platform for the in vivo assessment of putative disease-causing mutations in LQT genes, and of therapeutic interventions.

Posthumous diagnosis of long QT syndrome from neonatal screening cards.

BACKGROUND: Molecular autopsy in sudden unexplained death in the young (SUDY) victims cannot usually be performed if tissue suitable for DNA extraction is not retained at autopsy. OBJECTIVE: The purpose of this study was to assess the feasibility and clinical value of posthumous genetic testing for long QT syndrome (LQTS) using residual material from the neonatal screening (Guthrie) card in SUDY victims. METHODS: Twenty-one cases were investigated up to 13 years after death. Deaths occurred at <1 year in one, 1-18 years in 18, and 19-35 years in two patients. Guthrie cards were 3-39 years old. DNA was extracted, and amplicons corresponding to the coding regions of the LQTS genes 1, 2, 3, 5, and 6 underwent either denaturing high-performance liquid chromatography screening or direct DNA sequencing. RESULTS: Adequate DNA was extracted in every case, although repeated purification and amplification was often required. Rare variants were detected in six of 19 cases undergoing diagnostic screening. Four (21%) are considered to be pathological and have been used for family screening: R243C and H455Y in KCNQ1 in 12-year-old and 13-year-old boys, respectively, and Q81H and S621R in KCNH2 in 21-month and 28-year-old females, respectively. Variants of uncertain significance were R1047L in KCNH2 in a 2-year-old girl and S38G in KCNE1 in a 19-month-old boy. Point mutation tests for previously identified familial LQTS mutations revealed a positive result in both cases: E146K in KCNQ1 and exon 6-4del in KCNH2. CONCLUSION: Residual material from Guthrie cards collected for newborn metabolic screening can be used as a reliable source of DNA for the posthumous diagnosis of LQTS decades after SUDY, although purification and amplification of DNA is time intensive.

Echocardiographic assessment of blood flow volume in the superior vena cava and descending aorta in the newborn infant.

BACKGROUND: Clinical methods of assessing adequacy of the circulation are poor predictors of volume of blood flow in the newborn preterm. Doppler echocardiography can be used to assess perfusion at various sites in the circulation. OBJECTIVE: To assess repeatability of measurement of volume of superior vena caval (SVC) and descending aortic (DAo) flow. DESIGN: SVC and DAo flow volume were assessed four times in the first 48 h of postnatal life in a cohort of preterm (<31 weeks) infants. Within-observer and between-observer repeatability was assessed in a subgroup of preterm infants. Normative values were derived from 14 preterm infants who required <48 h respiratory support and 13 healthy term infants. RESULTS: Within-observer repeatability coefficient was 30 ml/kg/min for quantification of SVC flow, and 2.2 cm for DAo stroke distance. Measurement of DAo diameter had poor repeatability. Between-observer repeatability appeared poorer than within-observer repeatability. The fifth centile for volume of SVC flow in healthy preterm infants was 55 ml/kg/min and 4.5 cm for DAo stroke distance. CONCLUSIONS: Echocardiographic assessments of volume of SVC flow and velocity of DAo flow have similar within-observer repeatability to other neonatal haemodynamic measurements. Between-observer repeatability for both measurements was poor, reflecting the difficulty of standardising these novel techniques. In this small cohort of preterm infants, SVC flow volume <55 ml/kg/min and DAo stroke distance <4.5 cm represented low or borderline systemic perfusion in the first 48 h of postnatal life.

Relationship between blood pressure and blood flow in newborn preterm infants.

BACKGROUND: Arterial blood pressure remains the most frequently monitored indicator of neonatal circulatory status. However, studies of systemic perfusion in neonates have often shown only weakly positive associations with blood pressure. OBJECTIVES: To examine the relationship between invasively monitored arterial blood pressure and four measurements of systemic perfusion: left and right ventricular outputs, superior vena caval (SVC) flow and descending aortic (DAo) flow. DESIGN: Echocardiographic assessments of perfusion were performed four times in the first 48 h of postnatal life in a cohort of 34 preterm (<30 weeks) infants. Arterial blood pressure was monitored invasively over the exact duration of the echocardiogram. RESULTS: In the first 48 h of postnatal life there was no evidence of a positive association between blood pressure and volume of blood flow in any of the four vessels studied. At 5 h postnatal age there was a weak but significant inverse correlation between volume of SVC flow and arterial blood pressure (p = 0.04). A similar but non-significant trend was seen at 12 h postnatal age. CONCLUSIONS: Infants with reduced systemic perfusion tend to have normal or high blood pressure in the first hours of life, suggesting that a high systemic vascular resistance may lead to reduced blood flow. Low blood pressure does not correlate with poor perfusion in the first 48 h of postnatal life in sick preterm infants.

Neonatal tachycardias: an update.

This review provides an updated framework for the diagnosis and management of neonatal tachycardias.

Near-miss SIDS due to Brugada syndrome.

A 19 day old infant was successfully resuscitated from ventricular fibrillation. The 12 lead ECG was normal, with a normal QT interval, and remains so over three years follow up. DNA analysis revealed a missense mutation (R1193Q) in the SCN5A gene, previously linked with familial sudden unexpected nocturnal death syndrome, also known as Brugada syndrome.

Cardiorespiratory stability during echocardiography in preterm infants.

Blood pressure, heart rate, and oxygen saturation were monitored prospectively during 40 echocardiography recordings on 17 preterm infants (25-29 weeks; 510-1430 g), to examine whether echocardiography can be performed without disturbing cardiorespiratory status in preterm infants. There was no impact on absolute blood pressure. Heart rate increased by a mean of 4 beats per minute, and oxygen saturation decreased by a mean of 1% during echocardiography. While these changes reached statistical significance they are not of clinical significance as they remained well within ranges seen during control rest periods. All readings had greater minute-to-minute variability during echocardiography but differences were small and again remained within physiological ranges.

Use of the newborn screening card to define cause of death in a 12-year-old diagnosed with epilepsy.

Post-mortem investigation of sudden death in young people frequently reveals no overt cause for the death. Full investigation is hampered if tissue or blood is not retained for DNA analysis. We report a post mortem molecular diagnosis of long QT syndrome in a 12-year-old boy diagnosed with epilepsy who died suddenly playing sport. The DNA was extracted from an archived blood spot on his newborn screening ('Guthrie') card, which had been taken from him at 6 days of age. A missense mutation was detected in exon 5 of the KCNQ1 gene; R243C (835C > T), associated with long QT type 1. The same mutation was found in the mother (who now takes effective preventative therapy), but not in the sib who has now been reassured that she is not at risk of sudden death.

The repeatability of echocardiographic determination of right ventricular output in the newborn.

BACKGROUND: Non-invasive measurement of left ventricular output has been shown to be a repeatable technique. Little is known about the repeatability using echocardiography in determining pulmonary arterial diameters or right ventricular output. AIMS: To find the most repeatable point at which to measure pulmonary arterial diameter, and to compare the repeatability of determining right ventricular output with left ventricular output. METHODS: We assessed the Intra-observer and inter-observer repeatability for measuring the diameter of the pulmonary trunk in 24 term and 26 preterm infants, respectively. Interobserver repeatability was assessed for the diameters of the pulmonary trunk and aorta, for stroke distance, and for left and right ventricular output. RESULTS: The coefficients of variation for intra-observer repeatability were 4%, 7.5% and 9% respectively for measurements of the pulmonary valve, the pulmonary trunk, and the right ventricular outflow tract. There were significant differences between observers for measurement of the pulmonary trunk (p<0.001) and right ventricular outflow tract (p=0.011) but not for the pulmonary valve measured in either its long (p=0.22) or short axis (p=0.22). Significant differences between observers were also found for the pulmonary stroke distance measured in the long axis (p=0.004) and aortic diameter at end-diastole (p<0.001). The other parameters did not differ significantly and were used to calculate right and left ventricular output, respectively. Mean left ventricular output was 241 mls/kg/min, with mean differences between observers of 0.6 mls/kg/min (95% confidence interval (CI): -39.2 to 40.3 mls/kg/min). Mean right ventricular output was 255 mls/kg/min, with mean differences of 0.3 mls/kg/min (95% CI: -24.1 to 23.4 mls/kg/min). CONCLUSION: Measuring the diameter of the pulmonary trunk at the base of the valvar hinge points was most repeatable. Repeatability of right ventricular output was similar to that of left, with absolute values similar to those published by other workers.

Transcatheter patent ductus arteriosus occlusion: evolution of techniques and results from the 1990s.

OBJECTIVE: To review the evolution of transcatheter patent ductus arteriosus (PDA) occlusion techniques and results. METHODS: A single institution, retrospective review including all patients with intention to close a PDA from 1991 to 1998, with no exclusions. RESULTS: Rashkind occluder (n = 65), sideris double-button (n = 6), Cook detachable coil (n = 28) and Amplatzer ductal occluder (n = 4) were used. Successful implantation occurred in 99 of 103 patients. There was a need for a second transcatheter procedure to close residual ductal shunting in 12% of patients: Rashkind umbrellas (n = 8), double-button (n = 1), coils (n = 3). Eight patients (8%) required surgery, including 4 of 6 patients with the double-button occluder. CONCLUSIONS: The Rashkind occluder and the Sideris double-button device both had an unacceptably high rate of residual shunts requiring a second transcatheter procedure or surgical closure. Detachable coils and the Amplatzer ductal occluder have become the current technology of choice for transcatheter PDA closure with high success rates.

NADPH diaphorase and nitric oxide synthase in the corpus cardiacum-corpus allatum of the cockroach Diploptera punctata.

Juvenile hormone synthesis by corpora allata is regulated partly by allatostatin containing nerves from the brain that innervate the corpora cardiaca and the corpora allata. To investigate whether NO also participates in the regulation of juvenile hormone synthesis, antibody against NO synthase and the histochemical test for NADPH diaphorase activity, a marker for NO synthase, were applied to the corpora cardiaca-corpora allata of Diploptera punctata. Strong NADPH diaphorase activity occurred in corpus allatum cells but not in nerve fibers in the corpora allata or corpora cardiaca. In contrast, NO immunoreactivity occurred in nerves in the corpora cardiaca but not within the corpora allata. NO and allatostatin were not colocalized. NO synthase and NADPH diaphorase activity were localized in similar areas of the subesophageal ganglion and cells in the pars intercerebralis of the brain. Positive correlation of the quantity of NADPH diaphorase activity with juvenile hormone synthesis during the gonadotrophic cycle and lack of such correlation in subesophageal ganglia suggest that NADPH diaphorase activity reflects the necessity of NADPH in the pathway of juvenile hormone synthesis. These data suggest that NO is unlikely to play a significant role in the regulation of the corpora allata.

Development of a system to record cardiac output continuously in the newborn.

Intermittent recordings of Doppler flow velocity and cardiac output are of value during intensive care of the sick newborn infant but result in repeated disturbance of the child. We describe a new device for making continuous precordial recordings of Doppler flow velocity from the pulmonary artery in healthy resting newborn infants. Optimal probe siting was evaluated in six babies, and signals were found to be best when the pulmonary artery was insonated from the mid left parasternum. Continuous recordings were made in 13 other babies. Pulmonary artery velocities and, by calculation, cardiac output were measured continuously over periods ranging from 24 to 60 min. Median right ventricular output ranged widely from 148 to 246 mL x kg(-1) x min(-1). In contrast, for individual babies, the values were remarkably stable: the interquartile ranges varied from 13.2 to 29.9 mL x kg(-1) x min(-1). The simultaneous display of signal power allowed independent assessment of artifactual changes in cardiac output. This technique is feasible in healthy term infants and now requires evaluation in the intensive care setting where it may provide useful information concerning trends and short-term variability in right ventricular output.

Haemodynamic effects of altering arterial oxygen saturation in preterm infants with respiratory failure.

AIMS: To examine the haemodynamic effects of brief alteration in arterial oxygenation in preterm infants with respiratory failure. METHODS: Eighteen preterm infants with respiratory failure, aged 9-76 hours, underwent detailed Doppler echocardiographic assessment at 86%, 96%, and 100% SaO2, achieved by altering the FIO2. Sixteen were receiving intermittent positive pressure ventilation, median FIO2 0.45 (0.20-0.65), median mean airway pressure 12 cm H2O (0-20). SaO2 was stable for 15 minutes at each stage. Four parameters of pulmonary arterial pressure were measured: peak velocity of tricuspid regurgitation and peak velocity of left to right ductal flow, TPV:RVET ratio and PEP:RVET ratio, measured at the pulmonary valve, along with flow velocity integrals at the aortic and pulmonary valves, and systemic arterial pressure. Ductal size was graded into closed, small, moderate, large with imaging, pulsed and continuous wave Doppler. RESULTS: Between 86% and 96% SaO2, there were no consistent changes, but in three of the 12 with a patent ductus arteriosus (PDA) there was ductal constriction, with complete closure in one. Between 96% and 100% SaO2, peak ductal flow velocity rose significantly in four of eight with a PDA. Ductal constriction occurred in four infants; in three this was associated with a significant fall in aortic flow integral and a rise in aortic pressure (4-6 mm Hg). Overall, 11 infants went from 86% to 100% SaO2 and pulmonary arterial pressure fell significantly in seven. CONCLUSION: A brief rise in SaO2 within the range maintained by most neonatal units can cause significant ductal constriction. The fall in pulmonary arterial pressure with 100% SaO2 seen in most infants was associated with a fall in pulmonary blood flow (or no change), rather than a rise, indicating that the dominant haemodynamic effect was ductal constriction rather than pulmonary vasodilation.

Atrial fibrillation with neonatal pulmonary lymphangiectasia.

A female infant born at 28 weeks' gestation was found to have mild hydrops foetalis. Initial echocardiography showed a structurally normal heart. During the first week of life, episodic atrial tachycardia with 1:1 or 2:1 conduction was seen, requiring therapy with digoxin. The infant remained ventilator dependent, with a persistent, chylous pleural effusion which contained a preponderance of lymphocytes. Congenital pulmonary lymphangiectasia (CPL) was confirmed histologically. Worsening episodes of atrial tachycardia, including episodes of atrial fibrillation, were further investigated and a repeat echocardiogram revealed thickening of the entire right atrial wall. The cardiac findings of a thickened right atrial wall with the histological signs of myocarditis were thought to be the cause of paroxysms of atrial fibrillation, an extremely rare arrhythmia in the neonatal period. To the authors' knowledge there have been no previous reports of CPL in association with the cardiac abnormalities described herein.

Multicenter evaluation of the MicroScan Rapid Gram-Negative Identification Type 3 Panel.

The accuracy and performance of the revised MicroScan Rapid Gram-Negative Identification Type 3 Panel (Dade MicroScan Inc., West Sacramento, Calif.) were examined in a multicenter evaluation. The revised panel database includes data for 119 taxa covering a total of 150 species, with data for 12 new species added. Testing was performed in three phases: the efficacy, challenge, and reproducibility testing phases. A total of 405 fresh and stock gram-negative isolates comprising 54 species were tested in the efficacy phase; 96.8% of these species were identified correctly in comparison to the identification obtained either with the API 20E system (bioMérieux Vitek, Hazelwood, Mo.) or by the conventional tube method. The number of correctly identified isolates in the challenge phase, including new species added to the database, was 221 of 247, or 89.5%, in comparison to the number correctly identified by the conventional tube method. A total of 465 isolates were examined for intra- and interlaboratory identification reproducibility and gave an agreement of 464 of 465, or 99.8%. The overall reproducibility of each individual identification test or substrate was 14,373 of 14,384, or 99.9%. The new Rapid Gram-Negative Identification Type 3 Panel gave accurate and highly reproducible results in this multiple-laboratory evaluation.

Allatostatin in hemocytes of the cockroach Diploptera punctata.

Allatostatins are neuropeptides that inhibit the production, by the corpora allata, of a major insect hormone, juvenile hormone. These peptides are produced by cells of the brain and ganglia as well as by midgut endocrine cells. Transport from these sites may contribute to the allatostatin content in the hemolymph (insect blood). Using a monoclonal antibody against Diploptera punctata allatostatin I (A-P-S-G-A-Q-R-L-Y-G-F-G-L-NH2) and in situ hybridization with a digoxigenin-labeled cRNA probe generated from a portion of the allatostatin gene, it is demonstrated that allatostatin is present in and synthesized by granular hemocytes of D. punctata. About 5% of the hemocytes react with anti-allatostatin antibody and a similar number hybridize with a cRNA probe that detects allatostatin-specific mRNA. Electron micrographs showed that allatostatin-immunoreactive material occurs in membrane-bound, uniformly dense granules that frequently fill fusiform-shaped cells. Allatostatin in cell and plasma fractions of hemolymph quantified by enzyme-linked immunosorbent assay and by bioassay for inhibition of juvenile hormone synthesis in vitro indicated that about equal quantities (0.1-0.2 fmol/microl) are present in cell and plasma fractions. The production of allatostatin by hemocytes suggests that allatostatins may function as regulatory peptides in hemolymph activities in addition to their other known functions.