Non-invasive assessment of severe liver fibrosis in patients with Fontan-associated liver disease: The VALDIG-EASL FONLIVER cohort.

BACKGROUND & AIMS: Fontan-type surgery is used as a palliation for congenital heart disease with univentricular physiology but may, in the long term, lead to advanced chronic liver disease. This study assessed the accuracy of conventional non-invasive models in assessing liver fibrosis and introduces a new risk score employing non-invasive tools. METHODS: A prospective, cross-sectional, observational study was conducted across five European centers and encompassing all consecutive adult patients with Fontan circulation, liver biopsy and non-invasive tests (elastography, APRI, FIB-4, Fibrosis score, Doha, GUCI, and AAR). The primary outcome was the identification of severe liver fibrosis on biopsy. Multivariable logistic regression identified non-invasive predictors of severe fibrosis, leading to the development and internal validation of a new scoring model named the FonLiver risk score. RESULTS: In total, 217 patients (mean [standard deviation] age, 27.9 [8.9] years; 50.7% males) were included. Severe liver fibrosis was present in 47.9% (95% CI 41.2%-54.5%) and correlated with a lower functional class, protein-losing enteropathy, and compromised cardiopulmonary and systemic hemodynamics. The final FonLiver risk score incorporated liver stiffness measurement using transient elastography and platelet count and demonstrated strong discrimination and calibration (area under the receiver operating curve [AUROC] of 0.81). The FonLiver risk score outperformed conventional prediction models (APRI, FIB-4, Fibrosis score, Doha, GUCI, and AAR), which all exhibited worse performance in our cohort (AUROC < 0.70 for all). CONCLUSION: Severe liver fibrosis is prevalent in adults following Fontan-type palliation and can be effectively estimated using with the novel FonLiver risk score. This scoring system can be easily incorporated into the routine assessment of patients with Fontan circulation.

Effects of the lockdown due to the COVID-19 pandemic on alcohol consumption in patients under treatment in an alcohol relapse prevention programme. / Efectos del confinamiento en el hogar debido a la pandemia por COVID-19 en el consumo en pacientes en tratamiento en un programa de deshabituación de alcohol.

During the COVID-19 pandemic, several exceptional measures were put in place in order to avoid virus propagation, such as lockdown and the discontinuation of usual health care assistance services. It was considered that these changes might be associated with an increase in alcohol consumption and a higher risk of relapse for patients under treatment. The aim of this study was to assess changes in alcohol consumption during the lockdown period (between March and May, 2020) in patients following treatment under the Alcohol Use Disorders Programme at the "Hospital 12 de Octubre" in Madrid. A total of 311 patients were assessed through interviews carried out by telephone in accordance with usual clinical practice during that period. 76% of the total number of patients did not experience changes in their alcohol consumption, 9.2% stopped drinking and some experienced severe withdrawal syndrome, while 7.5% relapsed. The risk factors found for worsening the prognosis of the patients were: being female, drinking alcohol alone or at home, binge drinking, concomitant substance misuse and failure to attend therapy groups or self-help groups online during the lockdown. 31.6% of the sample described psychopathological symptoms due to the lockdown, especially those who already had psychiatric comorbidities. For this reason, we can conclude that during the lockdown as a result of the pandemic, most of our alcohol dependent patients did not modify their drinking patterns, but specific factors enabled us to identify a more vulnerable subgroup.

Durante la pandemia producida por la infección por el Covid-19 se produjeron una serie de cambios sociosanitarios excepcionales para evitar su propagación como el confinamiento en el hogar y la supresión de los servicios asistenciales sanitarios habituales. Se consideró que estos cambios podrían implicar un incremento en el consumo de alcohol y un mayor riesgo de recaídas para los pacientes en tratamiento. El objetivo de este estudio fue valorar los cambios en el consumo durante el período de confinamiento (marzo a mayo de 2020) en los pacientes en tratamiento en el programa de alcohol del Hospital Doce de Octubre de Madrid. Fueron valorados 311 pacientes mediante entrevista telefónica dentro de la práctica clínica habitual durante ese período. Un 76 % de los pacientes no presentaron cambios en su situación de consumo, un 9,2% de estos cesaron en el consumo, algunos de ellos con cuadros de abstinencia graves, y un 7,5% recayeron. El sexo femenino, el consumo en solitario o en el hogar, en atracón, o el de otras drogas de forma concomitante y el no estar en terapia grupal o no asistir a grupos de las asociaciones de ayuda mutua por videoconferencia durante el confinamiento fueron factores predictores de mal pronóstico. Un 31,6% presentó alteraciones psicopatológicas debidas al confinamiento, sobre todo, aquellos pacientes con comorbilidad psiquiátrica. Por lo tanto, en situaciones similares a esta, la mayoría de los pacientes en tratamiento no modifican el patrón de consumo, pero, ciertas características identifican un subgrupo de sujetos más vulnerables.

Technical modifications for transplant in the failing Fontan.

INTRODUCTION: Heart transplant after Fontan completion poses a unique surgical challenge. Twenty patients are presented, stressing the technical hints performed in the five anastomoses to match the graft in the recipient. METHODS: Data are collected from 20 Fontan patients between 2013 and 2019. Age (13 years), weight (37 kg.), and time interval between Fontan and transplant (7 years) are presented as median. Extracardiac conduit (size 18/20) was implanted in 15 patients, whereas atrio-pulmonary connection was performed in 4 and lateral tunnel in 1. Six patients developed protein-losing enteropathy. Seventeen stents had been previously deployed. RESULTS: The five anastomoses underwent some changes. Left atrium once, aorta 9 times, superior vena cava 7 times, pulmonary branches 15 times, and inferior vena cava 12 times. Follow-up was complete for a median of 42 months (range 6-84). Two patients died. ECMO was needed in six cases for pulmonary hypertension. Four patients had collateral vessels occluded in the cath lab, and stents were placed in superior vena cava (1) and aorta (1) post-transplant. Protein-losing enteropathy was resolved in five patients. Interestingly, one patient was on a systemic assist device before transplant (Levitronix) and right assistance (ECMO) afterwards. CONCLUSIONS: Transplant in Fontan patients is actually challenging. Hints in every of the five proposed anastomoses must be anticipated, including stents removal. Extra tissue from the donor (innominate vein, aortic arch, and pericardium) is strongly advisable. ECMO for right ventricular dysfunction was needed in nearly one-third of the cases. Overall results can match other transplant cohorts.

Prevalence, features and predictive factors of liver nodules in Fontan surgery patients: The VALDIG Fonliver prospective cohort.

BACKGROUND & AIMS: Fontan surgery is used to treat a variety of congenital heart malformations, and may lead to advanced chronic liver disease in the long-term. This study examines the prevalence, characteristics and predictors of liver nodules in patients following Fontan surgery. METHODS: This was a prospective, cross-sectional, observational study conducted at 8 European centres. Consecutive patients who had undergone Fontan surgery underwent blood tests, abdominal ultrasonography (US), transient elastography (Fibroscan®), echocardiography, haemodynamic assessments, and abdominal MRI/CT scan. The primary outcome measure was liver nodules detected in the MRI/CT scan. Predictors of liver nodules were identified by multivariate logistic regression. RESULTS: One hundred and fifty-two patients were enrolled (mean age 27.3 years). The mean time elapsed from surgery to inclusion was 18.3 years. Liver nodule prevalences were 29.6% (95% CI 23-37%) on US and 47.7% (95% CI 39-56%) on MRI/CT. Nodules were usually hyperechoic (76.5%), round-shaped (>80%), hyperenhancing in the arterial phase (92%) and located in the liver periphery (75%). The sensitivity and specificity of US were 50% (95% CI 38-62%) and 85.3% (95% CI 75-92%), respectively. Inter-imaging test agreement was low (adjusted kappa: 0.34). In the multivariate analysis, time since surgery >10 years was the single independent predictor of liver nodules (odds ratio 4.18; p = 0.040). Hepatocellular carcinoma was histologically diagnosed in 2 of the 8 patients with hypervascular liver nodules displaying washout. CONCLUSION: While liver nodules are frequent in Fontan patients, they may go unnoticed in US. Liver nodules are usually hyperechoic, hypervascular and predominantly peripheral. This population is at risk of hepatocellular carcinoma, the diagnosis of which requires confirmatory biopsy. LAY SUMMARY: Fontan surgery is the standard of care for many patients with univentricular congenital cardiopathies. Recent advances have improved the survival of Fontan patients, and nowadays most of them reach adulthood. In this setting, Fontan-associated liver disease (FALD) is increasingly recognised, and has become a significant prognostic factor. Liver nodules are considered a component of FALD yet their prevalence, imaging features and predictors have hardly been evaluated. In this study, we observed that liver nodules are frequent, typically hyperechoic, hypervascular and predominantly peripheral in patients with FALD. This population is at risk of hepatocellular carcinoma, the diagnosis of which must be confirmed by biopsy.

Inclusion of Alcoholic Associations Into a Public Treatment Programme for Alcoholism Improves Outcomes During the Treatment and Continuing Care Period: A 6-Year Experience.

AIMS: To investigate whether inclusion of self-help groups into the hospital treatment programme improves the prognosis of alcohol dependence through the treatment period; and to examine therapeutic adherence and prognosis during continuing care. METHOD: Patients attending the treatment programme at the 'Hospital 12 de Octubre' were randomized into two groups. In Group A (n = 123), patients received the usual treatment included in our programme, whereas in Group B (n = 126), patients also attended self-help groups. Patients were assessed with psychological scales at baseline, at the end of the treatment period and after completing the continuing care visits. Data were collected over a total of 6 years. RESULTS: During the treatment period, patients in Group B accumulated more months of abstinence and dropped out less. During continuing care, patients in Group B accumulated more months of abstinence and therapeutic adherence was higher. Variables that were associated with these results during the continuing care period were: visits to the GP, scores on anxiety, impulsivity and meaning of life scales, and belonging to the group that attended the alcoholic associations. CONCLUSIONS: Mutual help groups incorporated into a public treatment programme appear to improve outcomes during treatment and on into continuing care. This experience supports cooperation between public health centres and alcoholic associations in treating alcoholism. SHORT SUMMARY: Including alcoholic associations into the public treatment programme for alcoholism of the 'Hospital 12 de Octubre' in Madrid was shown to be associated with better outcomes in terms of months of accumulated abstinence, dropout rates and therapeutic adherence, during the treatment and continuing care periods.

Protecting the melatonin rhythm through circadian healthy light exposure.

Currently, in developed countries, nights are excessively illuminated (light at night), whereas daytime is mainly spent indoors, and thus people are exposed to much lower light intensities than under natural conditions. In spite of the positive impact of artificial light, we pay a price for the easy access to light during the night: disorganization of our circadian system or chronodisruption (CD), including perturbations in melatonin rhythm. Epidemiological studies show that CD is associated with an increased incidence of diabetes, obesity, heart disease, cognitive and affective impairment, premature aging and some types of cancer. Knowledge of retinal photoreceptors and the discovery of melanopsin in some ganglion cells demonstrate that light intensity, timing and spectrum must be considered to keep the biological clock properly entrained. Importantly, not all wavelengths of light are equally chronodisrupting. Blue light, which is particularly beneficial during the daytime, seems to be more disruptive at night, and induces the strongest melatonin inhibition. Nocturnal blue light exposure is currently increasing, due to the proliferation of energy-efficient lighting (LEDs) and electronic devices. Thus, the development of lighting systems that preserve the melatonin rhythm could reduce the health risks induced by chronodisruption. This review addresses the state of the art regarding the crosstalk between light and the circadian system.

Socioeconomic inequalities in COVID-19 incidence during the first six waves in Barcelona.

BACKGROUND: The emergence of SARS-CoV-2 affected urban areas. In Barcelona, six waves of COVID-19 hit the city between March 2020 and March 2022. Inequalities in the incidence of COVID-19 have been described. However, no studies have examined the daily trends of socioeconomic inequalities and how they changed during the different phases of the pandemic. The aim of this study is to analyse the dynamic socioeconomic inequalities in the incidence of COVID-19 during the six waves in Barcelona. METHODS: We examined the proportion of daily cases observed in the census tracts in the lower income tercile compared with the proportion of daily cases observed in the sum of the lower and higher income terciles. Daily differences in these proportions were assessed as a function of the epidemic waves, sex, age group, daily incidence and daily change in the incidence. A logistic regression model with an autoregressive term was used for statistical analysis. RESULTS: A time-dynamic effect was found for socioeconomic inequalities in the incidence of COVID-19. In fact, belonging to a lower-income area changed from being a risk factor (Waves 1, 2, 4 and 5) to being a protective factor in the sixth wave of the pandemic. Age also had a significant effect on incidence, which also changed over the different waves of the pandemic. Finally, the lower-income areas showed a comparatively lower incidence during the ascending phase of the epidemic waves. CONCLUSION: Socioeconomic inequalities in COVID-19 changed by wave, age group and wave phase.

Phase Response Curve to Light under Ambulatory Conditions: A Pilot Study for Potential Application to Daylight Saving Time Transitions.

Several studies have investigated the relationship between daylight saving time (DST) and sleep alterations, psychiatric disorders, cardiovascular events and traffic accidents. However, very few have monitored participants while maintaining their usual lifestyle before and after DST. Considering that DST transitions modify human behavior and, therefore, people's light exposure patterns, the aim of this study was to investigate the potential effects of DST on circadian variables, considering sleep and, for the first time, the human phase response curve to light. To accomplish this, eight healthy adults (33 ± 11 years old, mean ± SD) were recruited to monitor multivariable circadian markers and light exposure by means of a wearable ambulatory monitoring device: Kronowise®. The following night phase markers were calculated: midpoints of the five consecutive hours of maximum wrist temperature (TM5) and the five consecutive hours of minimum time in movement (TL5), sleep onset and offset, as well as sleep duration and light intensity. TM5 for wrist temperature was set as circadian time 0 h, and the balance between advances and delays considering the phase response curve to light was calculated individually before and after both DST transitions. To assess internal desynchronization, the possible shift in TM5 for wrist temperature and TL5 for time in movement were compared. Our results indicate that the transition to DST seems to force the circadian system to produce a phase advance to adapt to the new time. However, the synchronizing signals provided by natural and personal light exposure are not in line with such an advance, which results in internal desynchronization and the need for longer synchronization times. On the contrary, the transition back to ST, which implies a phase delay, is characterized by a faster adaptation and maintenance of internal synchronization, despite the fact that exposure to natural light would favor a phase advance. Considering the pilot nature of this study, further research is needed with higher sample sizes.

Correlated color temperature and light intensity: Complementary features in non-visual light field.

An appropriate exposure to the light-dark cycle, with high irradiances during the day and darkness during the night is essential to keep our physiology on time. However, considering the increasing exposure to artificial light at night and its potential harmful effects on health (i.e. chronodisruption and associated health conditions), it is essential to understand the non-visual effects of light in humans. Melatonin suppression is considered the gold standard for nocturnal light effects, and the activation of intrinsically photosensitive retinal ganglion cells (ipRGCs) through the assessment of pupillary light reflex (PLR) has been recently gaining attention. Also, some theoretical models for melatonin suppression and retinal photoreceptors activation have been proposed. Our aim in this study was to determine the influence of correlated color temperature (CCT) on melatonin suppression and PLR, considering two commercial light sources, as well as to explore the possible correlation between both processes. Also, the contribution of irradiance (associated to CCT) was explored through mathematical modelling on a wider range of light sources. For that, melatonin suppression and PLR were experimentally assessed on 16 healthy and young volunteers under two light conditions (warmer, CCT 3000 K; and cooler, CCT 5700 K, at ~5·1018 photons/cm2/sec). Our experimental results yielded greater post-stimulus constriction under the cooler (5700 K, 13.3 ± 1.9%) than under the warmer light (3000 K, 8.7 ± 1.2%) (p < 0.01), although no significant differences were found between both conditions in terms of melatonin suppression. Interestingly, we failed to demonstrate correlation between PLR and melatonin suppression. Although methodological limitations cannot be discarded, this could be due to the existence of different subpopulations of Type 1 ipRGCs differentially contributing to PLR and melatonin suppression, which opens the way for further research on ipRGCs projection in humans. The application of theoretical modelling suggested that CCT should not be considered separately from irradiance when designing nocturnal/diurnal illumination systems. Further experimental studies on wider ranges of CCTs and light intensities are needed to confirm these conclusions.

Electrochromic selective filtering of chronodisruptive visible wavelengths.

We present evidence of pupil response modification, as well as differential theoretical melatonin suppression through selective and dynamic electrochromic filtering of visible light in the 400-500 nm range to minimize chronodisruptive nocturnal blue light exposure. A lower activation of intrinsically photosensitive retinal ganglion cells (ipRGCs), the first step for light to reach a human's internal clock, is related to melatonin secretion therefore avoiding detrimental effects of excessive blue light exposure. Pupillary Light Reflex and Color Naming were experimentally assessed under light filtered by two different coloration states (transmissive and absorptive) of these novel dynamic filters, plus an uncoated test device, in 16 volunteers. Also, different commercial light sources at illuminances ranging from 1 to 1000 lux were differentially filtered and compared in terms of theoretical melatonin suppression. Representative parameters of the pupil responses reflected lower pupil constriction when the electrochromic filters (ECFs) were switched on (absorptive state, blue light is absorbed by the filter) compared to uncoated filters (control sample), but failed to do so under transmissive state (blue light passes through the filter) indicating less activation of ipRGCs under absorptive state (although no significant differences between states was found). Out of eight colors tested, just one showed significant differences in naming between both filter states. Thus, the ECF would have some protecting effect on ipRGC activation with very limited changes in color perception. While there are some limitations of the theoretical model used, the absorptive state yielded significantly lower theoretical melatonin suppression in all those light sources containing blue wavelengths across the illuminance range tested. This would open the way for further research on biological applications of electrochromic devices.

The impact factor: implications for research policy, editorial rules and scholarly reputation.

The assessment of scientific research is essentially based on several metric parameters, the so-termed Impact Factor perhaps being the predominant one. Despite well-founded criticisms and the wide opposition of reputed scientists, this procedure has become a tool of scientific policy, and is applied in editorial procedures for scientific publication, the evaluation of research groups, the concession of grants, fellowships or even academic positions. Indeed, cutting-edge research is today a competitive and exigent task, where the legitimacy and restrictions of such metric factors remain a preoccupation. However, whatever the policy of evaluation implemented, most breakthroughs are revolutionary, and involve a change in a given paradigm, usually being made by unorthodox scientists, whose scholarly reputation may be questioned by the establishment, and who may often be excluded as a result of the current system of highly productive research.

Determining Light Intensity, Timing and Type of Visible and Circadian Light From an Ambulatory Circadian Monitoring Device.

During last decades, the way of life in modern societies has deeply modified the temporal adjustment of the circadian system, mainly due to the inappropriate use of artificial lighting and the high prevalence of social jet-lag. Therefore, it becomes necessary to design non-invasive and practical tools to monitor circadian marker rhythms but also its main synchronizer, the light-dark cycle under free-living conditions. The aim of this work was to improve the ambulatory circadian monitoring device (ACM, Kronowise®) capabilities by developing an algorithm that allows to determine light intensity, timing and circadian light stimulation by differentiating between full visible, infrared and circadian light, as well as to discriminate between different light sources (natural and artificial with low and high infrared composition) in subjects under free living conditions. The ACM device is provided with three light sensors: (i) a wide-spectrum sensor (380-1100 nm); (ii) an infrared sensor (700-1100 nm) and (iii) a sensor equipped with a blue filter that mimics the sensitivity curve of the melanopsin photopigment and the melatonin light suppression curve. To calibrate the ACM device, different commercial light sources and sunlight were measured at four different standardized distances with both a spectroradiometer (SPR) and the ACM device. CIE S 026/E:2018 (2018), toolbox software was used to calculate the melanopic stimulation from data recorded by SPR. Although correlation between raw data of luminance measured by ACM and SPR was strong for both full spectrum (r = 0.946, p < 0.0001) and circadian channel (r = 0.902, p < 0.0001), even stronger correlations were obtained when light sources were clustered in three groups: natural, infrared-rich artificial light and infrared-poor artificial light, and their corresponding linear correlations with SPR were considered (r = 0.997, p < 0.0001 and r = 0.998, p < 0.0001, respectively). Our results show that the ACM device provided with three light sensors and the algorithm developed here allow an accurate detection of light type, intensity and timing for full visible and circadian light, with simultaneous monitoring of several circadian marker rhythms that will open the possibility to explore light synchronization in population groups while they maintain their normal lifestyle.

Valve area and the risk of overestimating aortic stenosis.

OBJECTIVE: To obtain reference values of aortic valve area (AVA) in a large population and to infer the risk of overestimating aortic stenosis (AS) when focusing on flow-corrected indices of severity. METHODS: We prospectively measured indices of AS in all consecutive echocardiograms performed in a large referral cardiac imaging laboratory for 1 year. We specifically analysed the distribution of AVA, indexed AVA and velocity ratio (Vratio) in patients with and without AS, the latter defined as the coexistence of valvular outflow obstruction (Vmax ≥2.5 m/s) and morphological findings of valve degeneration. RESULTS: 16 156 echocardiograms were analysed, 14 669 of which did not show valvular obstruction (peak jet velocity <2.5 m/s). In the latter group, AVA was 2.6±0.7 cm2 in 8190 studies with normal valves and 2.3±0.7 cm2 in 6479 studies with aortic sclerosis (AScl). There was a relatively wide overlap between values of AVA, indexed AVA and velocity ratio between studies of patients with AScl and AS. Values of AVA ≤1.0 cm2 were found in 0.5% of studies with normal valves and 1.8% of studies with AScl. These proportions were 3.1% and 9.3% for AVA ≤1.5 cm2, respectively. Vratio ≤0.25 were found in 0.1% of patients without obstruction. Risk factors for a small AVA in patients without obstruction were AScl, female sex, small body surface area, low ejection fraction and mitral regurgitation. CONCLUSIONS: Normal values of continuity-equation derived AVA are smaller than previously considered. AVA values below cutoffs of moderate or severe AS can be found in patients without the disease. Flow-corrected indices may overestimate AS in patients with low gradients, particularly in the presence of well-identified risk factors.

The Functional Significance of Paradoxical Low-Gradient Aortic Valve Stenosis: Hemodynamic Findings During Cardiopulmonary Exercise Testing.

OBJECTIVES: The goal of this study was to determine the functional impact of paradoxical low-gradient aortic stenosis (PLGAS) and clarify whether the relevance of the valvular obstruction is related to baseline flow. BACKGROUND: Establishing the significance of PLGAS is particularly challenging. METHODS: Twenty symptomatic patients (77 ± 6 years of age; 17 female subjects) with PLGAS (mean gradient 28 ± 6 mm Hg; aortic valve area 0.8 ± 0.1 cm2; ejection fraction 66 ± 7%) underwent cardiopulmonary exercise testing combined with right-heart catheterization and Doppler echocardiographic measurements. RESULTS: Aortic valve area increased by 84 ± 23% (p < 0.001) and, in 70% of subjects, it reached values >1.0 cm2 at peak exercise. Stroke volume index and blood pressure increased by 83 ± 56% and 26 ± 16%, respectively (both p < 0.0001). Peak oxygen consumption inversely correlated with the rate of increase in pulmonary capillary wedge pressure (PCWP) (PCWP slope: R = -0.61; p = 0.004). In turn, the PCWP slope was determined by changes in the valvular and vascular load but not by the rest of the indices of aortic stenosis. The functional impact of PLGAS was also not related to baseline flow. Agreement between Doppler echocardiography and the Fick technique was good up to intermediate workload. CONCLUSIONS: In symptomatic patients with PLGAS, the capacity to dynamically reduce vascular and valvular loads determines the effect of exercise on PCWP, which, in turn, conditions the functional status. A critically fixed valvular obstruction may not be the main mechanism of functional impairment in a large proportion of patients with PLGAS. Exercise echocardiography is suitable to study the dynamics of PLGAS.

Natural History and Clinical Predictors of Atrial Tachycardia in Adults With Congenital Heart Disease.

BACKGROUND: Atrial tachycardias (ATs) are a significant source of morbidity in adults with congenital heart disease (CHD). This study evaluates the incidence and clinical predictors of AT in a cohort of patients with CHD. METHODS AND RESULTS: We included 3311 adults (median age at entry 22.6 years, 50.6% males) with CHD (49% simple, 39% moderate, and 12% complex) prospectively followed up in a tertiary center for 37 607 person-years. Predictors of AT were identified by multivariable Cox regression analysis accounting for left truncation. An external validation was performed in a contemporary cohort of 1432 patients. Overall, 153 (4.6%) patients presented AT. AT burden was highest in complex CHD, such as single ventricle (22.8%) and d-transposition of the great arteries (22.1%). Hazard rates of AT across lifetime, age at presentation, and the time lapse between surgery and the first AT episode varied among the most common CHD. Independent risk factors for developing AT were univentricular physiology, previous intracardiac repair, systemic right ventricle, pulmonary hypertension, pulmonary regurgitation, pulmonary atrioventricular valve regurgitation, pulmonary and systemic ventricular dysfunction. At the age of 40 years, AT-free survival in patients with 0, 1, 2, and ≥3 risk factors was 100%, 94%, 76%, and 50%, respectively. These findings were confirmed in the validation cohort. CONCLUSIONS: Natural history of AT differed among the most common forms of CHD. Simple clinical parameters, easily obtained by noninvasive means, were independent predictors of AT in adults with CHD. Although risk was negligible in patients without any of these factors, their addition progressively increased AT burden.

Adenosine monophosphate is elevated in the bronchoalveolar lavage fluid of mice with acute respiratory toxicity induced by nanoparticles with high surface hydrophobicity.

Inhaled nanomaterials present a challenge to traditional methods and understanding of respiratory toxicology. In this study, a non-targeted metabolomics approach was used to investigate relationships between nanoparticle hydrophobicity, inflammatory outcomes and the metabolic fingerprint in bronchoalveolar fluid. Measures of acute lung toxicity were assessed following single-dose intratracheal administration of nanoparticles with varying surface hydrophobicity (i.e. pegylated lipid nanocapsules, polyvinyl acetate nanoparticles and polystyrene beads; listed in order of increasing hydrophobicity). Broncho-alveolar lavage (BAL) fluid was collected from mice exposed to nanoparticles at a surface area dose of 220 cm(2) and metabolite fingerprints were acquired via ultra pressure liquid chromatography-mass spectrometry-based metabolomics. Particles with high surface hydrophobicity were pro-inflammatory. Multivariate analysis of the resultant small molecule fingerprints revealed clear discrimination between the vehicle control and polystyrene beads (p < 0.05), as well as between nanoparticles of different surface hydrophobicity (p < 0.0001). Further investigation of the metabolic fingerprints revealed that adenosine monophosphate (AMP) concentration in BAL correlated with neutrophilia (p < 0.01), CXCL1 levels (p < 0.05) and nanoparticle surface hydrophobicity (p < 0.001). Our results suggest that extracellular AMP is an intermediary metabolite involved in adenine nucleotide-regulated neutrophilic inflammation as well as tissue damage, and could potentially be used to monitor nanoparticle-induced responses in the lung following pulmonary administration.

Development of a procedure for the isolation and enrichment of modified nucleosides and nucleobases from urine prior to their determination by capillary electrophoresis-mass spectrometry.

A sample treatment step based on solid-phase extraction (SPE) with polymeric sorbents has been developed for the simultaneous isolation and preconcentration of nucleosides and nucleobases from urine prior to analyses by CE-ESI-MS. In most reported methods nucleosides are isolated from urine by SPE in affinity mode, using an immobilized phenylboronic acid group, which specifically binds cis-diols. However, this is not applicable to non-cis-diol compounds. Here, different types of polymeric sorbents were evaluated for the simultaneous extraction of nucleosides and nucleobases from urine. The best results were obtained with Isolute ENV+, a hydroxylated styrene-divylbenzene polymer, whose retention capacity can be attributed mainly to hydrophobic interactions, and thus it can be applied to a broad range of compounds, regardless of whether they present or not to the cis-diol group in their structure. Other parameters such as the elution solvent and sample volume were optimized. We also studied the influence of the addition of isotopically labeled internal standards (ILISs) before or after the extraction step. The detection limits achieved were in the 0.04-0.17µg/mL range for a sample size of 2.0mL and relative standard deviations were 4-22%. The whole method developed, SPE prior to CE-ESI-MS, was applied to human urine samples from healthy volunteers. We conclude that SPE with polymeric sorbents prior to the electrophoretic CE-ESI-MS methodology constitutes a fast, valid and reliable approach for the simultaneously extraction of urinary nucleosides and nucleobases.

Capillary electrophoresis-mass spectrometry for direct determination of urinary modified nucleosides. Evaluation of synthetic urine as a surrogate matrix for quantitative analysis.

This work describes the development of a fast and reliable method based on capillary zone electrophoresis coupled with electrospray ionization-mass spectrometry (CZE-ESI-MS) for the determination of modified nucleosides in untreated human urine. The target compounds were guanine, 1-methyl-guanine, 7-methyl-guanine, 9-methyl-guanine, adenosine, 1-methyl-adenosine, cytidine, guanosine, 7-methyl-guanosine. As internal standards, ribose-2-(13)C-adenosine and 8-(13)C-guanine were used. The CZE separation was carried out in acidic medium (pH 2.5). MS detection with a single quadrupole, with ESI operating in positive-ion mode, was optimized. For the analysis of urine samples, owing to the endogenous character of these analytes different quantification strategies were explored. The standard additions method, matrix-matched calibration in synthetic urine and calibration in pure aqueous medium were compared in order to evaluate the endogenous levels of these compounds in human urine. The results obtained showed that calibration in synthetic urine as a surrogate matrix was an appropriate alternative to the method of standard additions for the accurate quantitation of compounds such as guanine, 1-methyl-guanine, 7-methyl-guanine, adenosine, 1-methyl-adenosine and cytidine by CE-ESI-MS directly in the urine matrix; values in the range 0.1µg/mL for cytidine and 6.4µg/mL for 7mGua, as the lowest and the highest level, were found in untreated urine from healthy volunteers. These results were confirmed by LC-MS/MS detection. It can be concluded that the electrophoretic CZE-ESI-MS methodology offers a valid and reliable alternative for the determination of urinary nucleosides at naturally occurring levels in healthy individuals.