Intraspinal microstimulation and diaphragm activation after cervical spinal cord injury.

Intraspinal microstimulation (ISMS) using implanted electrodes can evoke locomotor movements after spinal cord injury (SCI) but has not been explored in the context of respiratory motor output. An advantage over epidural and direct muscle stimulation is the potential of ISMS to selectively stimulate components of the spinal respiratory network. The present study tested the hypothesis that medullary respiratory activity could be used to trigger midcervical ISMS and diaphragm motor unit activation in rats with cervical SCI. Studies were conducted after acute (hours) and subacute (5-21 days) C2 hemisection (C2Hx) injury in adult rats. Inspiratory bursting in the genioglossus (tongue) muscle was used to trigger a 250-ms train stimulus (100 Hz, 100-200 µA) to the ventral C4 spinal cord, targeting the phrenic motor nucleus. After both acute and subacute injury, genioglossus EMG activity effectively triggered ISMS and activated diaphragm motor units during the inspiratory phase. The ISMS paradigm also evoked short-term potentiation of spontaneous inspiratory activity in the previously paralyzed hemidiaphragm (i.e., bursting persisting beyond the stimulus period) in ∼70% of the C2Hx animals. We conclude that medullary inspiratory output can be used to trigger cervical ISMS and diaphragm activity after SCI. Further refinement of this method may enable "closed-loop-like" ISMS approaches to sustain ventilation after severe SCI.NEW & NOTEWORTHY We examined the feasibility of using intraspinal microstimulation (ISMS) of the cervical spinal cord to evoke diaphragm activity ipsilateral to acute and subacute hemisection of the upper cervical spinal cord of the rat. This proof-of-concept study demonstrated the efficacy of diaphragm activation, using an upper airway respiratory EMG signal to trigger ISMS at the level of the ipsilesional phrenic nucleus during acute and advanced postinjury intervals.

Safety and quality assurance of chemotherapeutic preparations in a hospital production unit: acceptance sampling plan and economic impact.

OBJECTIVE: The opportunity to apply a sampling plan was evaluated. Costs were computed by a microcosting study. SETTING: In 2003, a sampling plan was defined to reduce the number of chemotherapy quality controls while preserving the same level of quality. Recent qualitative and quantitative changes led us to define a second sampling plan supplemented by an economic evaluation to determine the cost and cost-savings of quality control. METHODS: The study considers preparation produced during four semesters classified into three groups. The first one includes drugs produced below 200 batches a semester. Group 2, those for which the lot of preparation lots would have been rejected twice among these four semesters. Group 3, those would have been accepted (≥3 'acceptable lot'). A single sampling plan by attributes was applied to this group with an acceptance quality level of 1.65% and a lot tolerance percent defective below 5%. A micro-costing study was conducted on quality control, from the sampling to the validation of the results. RESULTS: Among 39 cytotoxic drugs, 11 were sampled which enabled to avoid a mean of 17,512 control assays per year. Each batch of the 28 non-sampled drugs was however analyzed. Costs were estimated at 2.98€ and 5.25€ for control assays depending of the analytical method. The savings from the application of the sampling plans was 153,207€ in 6 years. CONCLUSION: The sampling plan allowed maintaining constancy in number of controls and the level of quality with significant costsavings, despite a substantial increase in drugs to assay and in the number of preparations produced.

Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus.

This study explored whether Crassostrea gigas oysters can be used as a bioindicator of white spot syndrome virus (WSSV) in shrimp farm water canals. Bioassays showed that C. gigas can accumulate WSSV in their gills and digestive glands but do not become infected, either by exposure to seawater containing WSSV or by cohabitation with infected shrimp. The use of a WSSV nested PCR to screen oysters placed in water canals at the entry of a shrimp farm allowed WSSV to be detected 16 d prior to the disease occurring. The finding that C. gigas can concentrate small amounts of WSSV present in seawater without being harmed makes it an ideal sentinel species at shrimp farms.

Perfusion quantification in contrast-enhanced ultrasound (CEUS)--ready for research projects and routine clinical use.

With contrast-enhanced ultrasound (CEUS) now established as a valuable imaging modality for many applications, a more specific demand has recently emerged for quantifying perfusion and using measured parameters as objective indicators for various disease states. However, CEUS perfusion quantification remains challenging and is not well integrated in daily clinical practice. The development of VueBox™ alleviates existing limitations and enables quantification in a standardized way. VueBox™ operates as an off-line software application, after dynamic contrast-enhanced ultrasound (DCE-US) is performed. It enables linearization of DICOM clips, assessment of perfusion using patented curve-fitting models, and generation of parametric images by synthesizing perfusion information at the pixel level using color coding. VueBox™ is compatible with most of the available ultrasound platforms (nonlinear contrast-enabled), has the ability to process both bolus and disruption-replenishment kinetics loops, allows analysis results and their context to be saved, and generates analysis reports automatically. Specific features have been added to VueBox™, such as fully automatic in-plane motion compensation and an easy-to-use clip editor. Processing time has been reduced as a result of parallel programming optimized for multi-core processors. A long list of perfusion parameters is available for each of the two administration modes to address all possible demands currently reported in the literature for diagnosis or treatment monitoring. In conclusion, VueBox™ is a valid and robust quantification tool to be used for standardizing perfusion quantification and to improve the reproducibility of results across centers.

The effect of fire frequency on local cembra pine populations.

It has been predicted that global climate change will lead to increasing drought in the Alps during the 21st century, as well as an increased fire risk, fires being currently rare in these mountains. Herein we describe fire frequency reconstruction using high-resolution analyses of macroscopic sedimentary charcoal, pollen, and plant macrofossils. Sediments were sampled from a subalpine pond within the dry western French Alps. Results show that the early-Holocene expansion of Pinus cembra (7200 calibrated years BP) occurred in Acer/Alnus incana/Betula woodlands, which were affected by fires with moderate mean fire-free intervals (MFFI = 173 +/- 61 yr [mean +/- SE]). Superposed Epoch Analyses show that the abundance of P. cembra macroremains decreased significantly after burning, although they never disappeared entirely. Statistics suggest that fires spread through cembra pine communities; they were not stand-replacing fires but mainly surface fires, probably killing nonreproductive pines. An increase in fire frequency occurred 6740 years ago, when four fires appear to have occurred within 140 years. These fires may have been associated with a regional drought and could have affected the composition of the subalpine forest by depleting the local P. cembra population in the short-term. The predicted increase in drought in the future could, therefore, affect the cembra pine ecosystem in the Alps if fire frequency is reduced to intervals of less than 80 years.

A beta-1,3 glucan sulfate induces resistance in grapevine against Plasmopara viticola through priming of defense responses, including HR-like cell death.

Sulfated laminarin (PS3) has been shown previously to be an elicitor of plant defense reactions in tobacco and Arabidopsis and to induce protection against tobacco mosaic virus. Here, we have demonstrated the efficiency of PS3 in protecting a susceptible grapevine cultivar (Vitis vinifera cv. Marselan) against downy mildew (Plasmopara viticola) under glasshouse conditions. This induced resistance was associated with potentiated H2O2 production at the infection sites, upregulation of defense-related genes, callose and phenol depositions, and hypersensitive response-like cell death. Interestingly, similar responses were observed following P. viticola inoculation in a tolerant grapevine hybrid cultivar (Solaris). A pharmacological approach led us to conclude that both callose synthesis and jasmonic acid pathway contribute to PS3-induced resistance.

Organoids as preclinical models to improve intraperitoneal chemotherapy effectiveness for colorectal cancer patients with peritoneal metastases: Preclinical models to improve HIPEC.

BACKGROUND: Peritoneal metastases (PM), corresponding to tumor implants into the peritoneal cavity, are associated with impaired prognosis and low responsiveness to systemic chemotherapy. A new therapeutic approach has dramatically changed the prognosis of patients with PM from colorectal cancer (CRC), consisting in the association of a complete cytoreductive surgery followed by intraperitoneal chemotherapy associated to hyperthermia (HIPEC). Many drugs have been administered intraperitoneally, but no clear consensus has been approved. Therefore, relevant preclinical models are essentials for the efficient translation of treatments option into affected patients. METHOD: Organoids, the last generation of preclinical models, were used to rationalize and improve intraperitoneal chemotherapy. We tested several cytotoxics, combination, effect of hyperthermia, exposure duration and frequency. RESULTS: Organoids were a representative model of response to chemotherapies used for the treatment of PM from CRC; 460mg/m2 of oxaliplatin being the most efficient cytotoxic treatment. Repeated incubations with oxaliplatin; mimicking cycles of intraperitoneal treatment, resulted in an increased efficacy. CONCLUSION & DISCUSSION: Organoids are relevant models to study the chemosensitivity of peritoneal metastases from CRCs. These models could be used for large scale drug screening strategies or personalized medicine, for colorectal carcinoma but also for PM from other origins.

Near infrared spectroscopy and process analytical technology to master the process of busulfan paediatric capsules in a university hospital.

The prescription of unlicensed oral medicines in paediatrics leads the hospital pharmacists to compound hard capsules, such as busulfan, an alkylating agent prescribed in preparative regimens for bone marrow transplantation. In this study, we have investigated how the general principle of process analytical technology (PAT) can be implemented at the small size of our hospital pharmacy manufacturing unit. Near infrared spectroscopy (NIRS) was calibrated for raw material identification, blend uniformity analysis and final content uniformity of busulfan hard capsules of 11 different strengths. Measurements were performed on capsules from 2 to 40 mg (n=440). After optimisation, accuracy and linearity of the NIRS quantitative method was demonstrated after comparison with a previously validated quantitative high performance thin layer chromatography (HPTLC) method. Such a comparison led to attractive NIRS precision: +/-0.7 to +/-1.0 mg for capsules from 2 to 40 mg, respectively. As NIRS is a rapid and non-destructive technique, the individual control of a whole batch of busulfan paediatric capsules intended to be administrated is possible. Actually, mastering the process of busulfan paediatric capsules with the NIRS integrated into the notion of PAT is a powerful analytical tool to assess the process quality and to perform content uniformity of at least 5mg busulfan-containing capsules.

Differential modulation of gene induction by glucocorticoids and antiglucocorticoids in rat hepatoma tissue culture cells.

Studies of glucocorticoid and antiglucocorticoid induction of tyrosine aminotransferase (TAT) in two rat hepatoma cell lines (Fu5-5 and HTC) are described. These studies revealed several phenomena that are not consistent with the current models of steroid hormone action: (a) TAT induction occurred at glucocorticoid levels below those required for comparable receptor occupancy in Fu5-5, but not in HTC, cells; (b) the ability of antiglucocorticoids to induce TAT is higher in Fu5-5 than in HTC cells; (c) the values of the amount of TAT agonist activity with the antiglucocorticoid dexamethasone 21-mesylate and of log10 of the dexamethasone concentration required for half-maximal induction of TAT were not constant over time but varied in a linear, reciprocal manner. This modulation was seen for several glucocorticoids and antiglucocorticoids at the level of both TAT enzyme and mRNA but not for two other glucocorticoid inducible genes in the same cells. These results, plus the fact that a similar difference in the concentration required for half-maximal TAT induction in Fu5-5 cells was seen for both glucocorticoids and cyclic AMP, argue that the modulation occurs at some point distal to receptor-steroid complex binding to the biologically active nuclear sites but proximal to translation of TAT mRNA. In order to explain these results, it is pointed out that models involving second messengers are entirely appropriate for steroid hormone action. The participation of a modulated trans-acting factor in such a model may explain the above results.

Prospective multicenter study of pregnancy outcomes in women with heart disease.

BACKGROUND: The maternal and neonatal risks associated with pregnancy in women with heart disease receiving comprehensive prenatal care have not been well defined. METHODS AND RESULTS: We prospectively enrolled 562 consecutive pregnant women with heart disease and determined the outcomes of 599 pregnancies not ending in miscarriage. Pulmonary edema, arrhythmia, stroke, or cardiac death complicated 13% of pregnancies. Prior cardiac events or arrhythmia, poor functional class or cyanosis, left heart obstruction, and left ventricular systolic dysfunction independently predicted maternal cardiac complications; the cardiac event rate can be predicted using a risk index incorporating these predictors. Neonatal complications (20% of pregnancies) were associated with poor functional class or cyanosis, left heart obstruction, anticoagulation, smoking, and multiple gestations. CONCLUSIONS: Pregnancy in women with heart disease is associated with significant cardiac and neonatal complications, despite state-of-the-art obstetric and cardiac care. Maternal cardiac risk can be predicted with the use of a risk index.

Simultaneous determination of aortic valve area by the Gorlin formula and by transesophageal echocardiography under different transvalvular flow conditions. Evidence that anatomic aortic valve area does not change with variations in flow in aortic stenosis.

OBJECTIVES: The purpose of this study was to determine the impact of changes in flow on aortic valve area (AVA) as measured by the Gorlin formula and transesophageal echocardiographic (TEE) planimetry. BACKGROUND: The meaning of flow-related changes in AVA calculations using the Gorlin formula in patients with aortic stenosis remains controversial. It has been suggested that flow dependence of the calculated area could be due to a true widening of the orifice as flow increases or to a disproportionate flow dependence of the formula itself. Alternatively, anatomic AVA can be measured by direct planimetry of the valve orifice with TEE. METHODS: Simultaneous measurement of the planimetered and Gorlin valve area was performed intraoperatively under different hemodynamic conditions in 11 patients. Left ventricular and ascending aortic pressures were measured simultaneously after transventricular and aortic punctures. Changes in flow were induced by dobutamine infusion. Using multiplane TEE, AVA was planimetered at the level of the leaflet tips in the short-axis view. RESULTS: Overall, cardiac output, stroke volume and transvalvular volume flow rate ranged from 2.5 to 7.3 liters/min, from 43 to 86 ml and from 102 to 306 ml/min, respectively. During dobutamine infusion, cardiac-output increased by 42% and mean aortic valve gradient by 54%. When minimal flow was compared with maximal flow, the Gorlin area varied from (mean +/- SD) 0.44 +/- 0.12 to 0.60 +/- 0.14 cm2 (p < 0.005). The mean change in Gorlin area under different flow rates was 36 +/- 32%. Despite these changes, there was no significant change in the planimetered area when minimal flow was compared with maximal flow. The mean difference in planimetered area under different flow rates was 0.002 +/- 0.01 cm2 (p = 0.86). CONCLUSIONS: By simultaneous determination of Gorlin formula and TEE planimetry valve areas, we showed that acute changes in transvalvular volume flow substantially altered valve area calculated by the Gorlin formula but did not result in significant alterations of the anatomic valve area in aortic stenosis. These results suggest that the flow-related variation in the Gorlin AVA is due to a disproportionate flow dependence of the formula itself and not a true change in valve area.

Dissociation of steroid binding to receptors and steroid induction of biological activity in a glucocorticoid-responsive cell.

Glucocorticoid responses in two independently derived lines of rat hepatoma tissue culture cells (HTC and FU5-5) were examined. FU5-5 cells exhibited induction of the enzyme tyrosine aminotransferase (TAT) at concentrations of dexamethasone that were approximately 7-fold lower than that required for HTC cells. FU5-5 cells also displayed substantial TAT induction with steroids that were partial agonists, or antagonists, in HTC cells. The increased sensitivity of FU5-5 cells was not, however, due to an increased affinity of FU5-5 cell receptors for dexamethasone, as determined from cell-free and whole cell binding experiments. The differential steroid sensitivity for TAT induction was observed with three other, structurally different glucocorticoids, thus apparently ruling out steroid metabolism in one of the cell lines as a cause. Also, induction of TAT in FU5-5 cells occurred at approximately 9-fold lower steroid concentrations than were required for the induction of glutamine synthetase (GS) in the same cells. Thus, the dose-response curves for TAT induction in HTC cells and for GS induction in FU5-5 cells are closely correlated with the saturation curve for whole cell steroid binding to receptor sites, while the dose-response curve for TAT induction in FU5-5 cells is shifted to lower steroid concentrations. This represents the first report of dissociation of two supposedly primary, glucocorticoid-induced functions and indicates that identical receptor-mediated processes cannot be utilized by FU5-5 cells for the induction of TAT and GS. The involvement of second messengers or different nuclear processes are possible explanations for the unusual behavior of FU5-5 cells during glucocorticoid induction of TAT.

Glucocorticoid versus antiglucocorticoid activity: can a single functional group modification of glucocorticoid steroids always convey antiglucocorticoid activity?

Single functional group modifications of glucocorticoid steroids have been performed in an effort to obtain antiglucocorticoids with high affinity and specificity for glucocorticoid receptors. This approach tests the hypothesis that the structural determinants of biological activity and receptor binding are independent so that modification of more potent glucocorticoids could yield more potent antiglucocorticoids . In this study, a new functional group capable of conferring antiglucocorticoid activity has been identified, i.e. the spiro C-17 oxetan -3'-one group. Using three glucocorticoids of greatly different potency ( deacylcortivazol greater than dexamethasone greater than cortisol), we examined the effects of incorporation of the oxetanone group and the previously described, alkylating C-21 mesylate group on steroid affinity for receptors and biological activity. In both series of modified steroids, the receptor affinity of the derivatives paralleled that of the parent steroids. The biological activities of the dexamethasone and cortisol derivatives were predominantly or totally antagonistic, while both deacylcortivazol derivatives were full agonists. We conclude that antiglucocorticoid activity can arise from the incorporation of a single functional group into glucocorticoid steroid structures, but that the expression of agonist vs. antagonist activity is determined by a balance of structural group determinants which are not restricted to a common region of the steroid. Within a given class of derivatives, receptor affinity correlated with the amount of agonist activity. The structure-activity relationships for dexamethasone oxetanone and deacylcortivazol mesylate were studied in detail. Dexamethasone oxetanone is a potent antiglucocorticoid in HTC cells. [3H]Dexamethasone oxetanone binds to cell-free glucocorticoid receptors with a Kd of 3.2 X 10(-8) M. No specific antiglucocorticoid binder was detected. Direct binding experiments with [3H]dexamethasone oxetanone as well as indirect studies of the kinetics of cell-free competition of [3H]dexamethasone binding demonstrated that dexamethasone oxetanone binds to receptors faster (by about a factor of 2) and dissociates from receptors much faster than does dexamethasone. Deacylcortivazol mesylate was a more potent agonist and binder to receptors than dexamethasone, but displayed no irreversible interactions with HTC cell receptors under those conditions that afforded a covalent receptor-steroid complex with the closely related dexamethasone mesylate.(ABSTRACT TRUNCATED AT 400 WORDS)

Impact of motor, cognitive, and perceptual disorders on ability to perform activities of daily living after stroke.

BACKGROUND AND PURPOSE: Using confirmatory factor analysis, this study evaluates the relative impact of motor, cognitive, and perceptual deficits on functional autonomy with 100 elderly (aged 55 to 79 years) victims of stroke. METHODS: Two different approaches were used for measuring functional autonomy: the Functional Autonomy Measurement System (Système de Mesure de l'Autonomie Fonctionnelle [SMAF]) and the Assessment of Motor and Process Skills (AMPS). RESULTS: The results of the confirmatory factor analysis show that motor, cognitive, and perceptual factors all make a significant contribution to the variation in functional autonomy and confirm the accuracy of the model (93% of the variance is explained when the SMAF is used to measure functional autonomy, and 64% of the variance is explained when the AMPS is used). CONCLUSIONS: The factors that make the greatest contribution in explaining the variance in functional autonomy are, in order of importance, the motor factor, the perceptual factor, and the cognitive factor.

Molecular analysis of RAPD DNA based markers: their potential use for the detection of genetic variability in jojoba (Simmondsia chinensis L Schneider).

We have applied the recently developed technique of random amplified polymorphic DNA (RAPD) for the discrimination between two jojoba clones at the genomic level. Among a set of 30 primers tested, a simple reproducible pattern with three distinct fragments for clone D and two distinct fragments for clone E was obtained with primer OPB08. Since RAPD products are the results of arbitrarily priming events and because a given primer can amplify a number of non-homologous sequences, we wondered whether or not RAPD bands, even those of similar size, were derived from different loci in the two clones. To answer this question, two complementary approaches were used: i) cloning and sequencing of the amplification products from clone E; and ii) complementary Southern analysis of RAPD gels using cloned or amplified fragments (directly recovered from agarose gels) as RFLP probes. The data reported here show that the RAPD reaction generates multiple amplified fragments. Some fragments, although resolved as a single band on agarose gels, contain different DNA species of the same size. Furthermore, it appears that the cloned RAPD products of known sequence that do not target repetitive DNA can be used as hybridization probes in RFLP to detect a polymorphism among individuals.

[Characterization of specific receptors for estradiol, induction of vitellogenin and its mRNA in the liver of rainbow trout (Salmo gairdnerii)]. / Caractérisation de récepteurs spécifiques à l'oestradiol, induction de la vitellogénine et de son mRNA dans le foie de truite arc-en-ci el (Salmo gairdnerii).

In all egg laying vertebrates, synthesis and use of vitellogenin (Vg) are intimately bound to the active phase of reproduction. In the liver of the rainbow trout (Salmo gairdnerii), Vg synthesis is influenced by estradiol (E2) which, we believe, acts through the classical mechanism of steroid hormone action. After binding of the hormone to a soluble specific receptor protein, the estradiol-receptor complex can interact with chromatin and modulate the expression of Vg genes, leading to increased synthesis of specific mRNA and Vg. We show here: (i) the presence of specific oestrogen receptors (dissociation constant KD congruent to 1.5 X 10(-9) M for E2) in the cytosol of the male trout liver. (ii) The male liver, offering, an ideal experimental control of "zero" background, we followed-in the liver of male trout--the kinetics of induction of Vg mRNA by hybridization with Vg cDNA, after E2 stimulation, and (iii) the apparition of Vg in the serum by using an original rocket immuno-electrophoretic technique. The male trout liver vitellogenin model and the original techniques we developed will be very useful to study the influence of endogenous and exogenous factors on the different steps (receptors, transcription, translation) of vitellogenesis regulation.

Early increases in RNA polymerase I activity and 18S and 28S rRNA synthesis in the male rat pituitary after oestradiol treatment.

This article reports the effect of a single injection of 17 beta-oestradiol on RNA synthesis, in the male rat pituitary. An increase in RNA polymerase I activity, with a maximum effect between 10 and 15 hours, is described. No modification in RNA polymerase II activity was detected. These results were extended and confirmed, using in vitro double labelling of RNA, following in vivo oestrogen treatment. Polyacrylamide gel electrophoresis of nuclear and cytoplasmic RNA showed an increased incorporation of adenine into 28S and 18S rRNA, in the pituitaries of oestrogen-treated animals. The 5S rRNA was not modified by the hormonal treatment. These effects on RNA polymerase I activity and on 28S and 18S rRNA synthesis were closely correlated with the long-term nuclear retention of receptor-oestradiol complexes, in vivo. Taken together, these observations argue in favor of the nucleolus as a preferential target for receptor-bound oestradiol, in the cell nucleus of the male rat pituitary.

A simple method for the preparation of the covalently closed circular form of plasmid DNA.

We describe a simple, rapid, inexpensive method for isolation of covalently closed circular plasmid DNA. The method involves the electrophoresis of crude DNA preparations in an agarose gel, electrotransfer onto a dialysis membrane and elution of the highly purified circular covalently closed plasmid DNA. Native and recombinant plasmid DNA have been purified by this method and shown to be suitable for restriction enzyme digestion and transformation of bacteria. The yield of this rapid purification procedure makes it a good alternative method to standard centrifugation in cesium chloride ethidium bromide gradients.

Doppler echocardiographic predictors of recurrence of atrial fibrillation after cardioversion.

To determine if the return of atrial contraction as evidenced by progressive return of the A wave on the Doppler atrial profile could better predict long-term success of cardioversion than other clinical and echocardiographic parameters, 50 patients were studied 4 hours, 24 hours and, if sinus rhythm persisted, up to 180 days after cardioversion. Recurrence of atrial fibrillation (AF) was 64% at 6 months. Age, sex, prior episodes of AF, presence of mitral valve disease or magnitude of mitral valve gradient did not predict recurrence, but duration of AF was significantly longer in the failure group (p less than 0.01). Left atrial dimension greater than or equal to 45 mm had a positive predictive value of 66%, with a sensitivity of 59% and a specificity of 61%. Presence or magnitude of the A wave at 4 hours did not predict long-term success of cardioversion. Percent increase of the A wave from 4 to 24 hours less than 10% had the highest positive predictive value (80%) for recurrence of AF (sensitivity 71% and specificity 71%) and can be obtained in the immediate post-cardioversion period to better establish prognosis and adjust therapeutic regimens.

Outcome of children with atrial septal defect considered too small for surgical closure.

There are few studies providing information on the natural course of hemodynamically insignificant atrial septal defect (ASD). To review the outcome of patients with secundum ASD, we retrospectively reviewed the charts of patients who had initially not been considered for surgical closure after age 1 year, and who had either a follow-up of at least 10 years or documented closure. Thirty patients, 22 females and 8 males, fulfilled our inclusion criteria. Mean age at diagnosis was 1.3 year and mean follow-up duration was 11.5 years. Seventeen patients had spontaneous closure of the ASD at a mean age of 8.4 years. There were 7 asymptomatic patients whose ASD was still patent at the last visit (mean age 14.1 years, mean follow-up 13.2), with defect dimensions on echocardiography ranging from 1 to 6 mm. The remaining 6 patients were considered to require surgical closure on the basis of an apparent increase in size of the ASD and secondary clinical and hemodynamic manifestations. These results (1) confirm that not all secundum ASDs need to be treated surgically because they can still spontaneously close past the age of 5, and (2) suggest that in a minority of cases the size of the defect could increase.