Effects of cytosine methylation on DNA morphology: An atomic force microscopy study.

Methylation is one of the most important epigenetic mechanisms in eukaryotes. As a consequence of cytosine methylation, the binding of proteins that are implicated in transcription to gene promoters is severely hindered, which results in gene regulation and, eventually, gene silencing. To date, the mechanisms by which methylation biases the binding affinities of proteins to DNA are not fully understood; however, it has been proposed that changes in double-strand conformations, such as stretching, bending, and over-twisting, as well as local variations in DNA stiffness/flexibility may play a role. The present work investigates, at the single molecule level, the morphological consequences of DNA methylation in vitro. By tracking the atomic force microscopy images of single DNA molecules, we characterize DNA conformations pertaining to two different degrees of methylation. In particular, we observe that methylation induces no relevant variations in DNA contour lengths, but produces measurable incremental changes in persistence lengths. Furthermore, we observe that for the methylated chains, the statistical distribution of angles along the DNA coordinate length is characterized by a double exponential decay, in agreement with what is predicted for polyelectrolytes. The results reported herein support the claim that the biological consequences of the methylation process, specifically difficulties in protein-DNA binding, are at least partially due to DNA conformation modifications.

Biophysical characterization of a binding site for TLQP-21, a naturally occurring peptide which induces resistance to obesity.

Recently, we demonstrated that TLQP-21 triggers lipolysis and induces resistance to obesity by reducing fat accumulation [1]. TLQP-21 is a 21 amino acid peptide cleavage product of the neuroprotein VGF and was first identified in rat brain. Although TLQP-21 biological activity and its molecular signaling is under active investigation, a receptor for TLQP-21 has not yet been characterized. We now demonstrate that TLQP-21 stimulates intracellular calcium mobilization in CHO cells. Furthermore, using Atomic Force Microscopy (AFM), we also provide evidence of TLQP-21 binding-site characteristics in CHO cells. AFM was used in force mapping mode equipped with a cantilever suitably functionalized with TLQP-21. Attraction of this functionalized probe to the cell surface was specific and consistent with the biological activity of TLQP-21; by contrast, there was no attraction of a probe functionalized with biologically inactive analogues. We detected interaction of the peptide with the binding-site by scanning the cell surface with the cantilever tip. The attractive force between TLQP-21 and its binding site was measured, statistically analyzed and quantified at approximately 40 pN on average, indicating a single class of binding sites. Furthermore we observed that the distribution of these binding sites on the surface was relatively uniform.

A case of spontaneous gastric rupture in a 5 years girl.

We describe a case of spontaneous gastric rupture in a child of 5 years old. The patient reached us in a serious condition; the anamnesis was negative for traumatic events or gastrointestinal disorders. An abdominal X-ray and CT scan revealed free air and fluid in the abdominal cavity, leading to the diagnosis of gastro-intestinal perforation. Submitted to urgent surgery, a rupture of the posterior wall of the stomach was found that was treated with gastrectomy "à la demande". The surgery follow-up was regular. Morphological and immunohistochemical study showed some muscular abnormalities of the muscular gastric wall.

Thermodynamic relation between voltage-concentration dependence and salt adsorption in electrochemical cells.

Electrochemical cells containing two electrodes dipped in an ionic solution are widely used as charge accumulators, either with polarizable (supercapacitor) or nonpolarizable (battery) electrodes. Recent applications include desalination ("capacitive deionization") and energy extraction from salinity differences ("capacitive mixing"). In this Letter, we analyze a general relation between the variation of the electric potential as a function of the concentration and the salt adsorption. This relation comes from the evaluation of the electrical and mechanical energy exchange along a reversible cycle, which involves salt adsorption and release by the electrodes. The obtained relation thus describes a connection between capacitive deionization and capacitive mixing. We check this relation with experimental data already reported in the literature, and moreover by some classical physical models for electrodes, including polarizable and nonpolarizable electrodes. The generality of the relation makes it very useful in the study of the properties of the electric double layer.

Single-molecule study of the DNA denaturation phase transition in the force-torsion space.

We use the "magnetic tweezers" technique to show the structural transitions that the DNA undergoes in the force-torsion space. In particular, we focus on the regions corresponding to negative supercoiling. These regions are characterized by the formation of the so-called denaturation bubbles, which play an essential role in the replication and transcription of DNA. We experimentally map the region of the force-torsion space where the denaturation takes place. We observe that large fluctuations in DNA extension occur at one of the boundaries of this region, i.e., when the formation of denaturation bubbles and of plectonemes compete. To describe the experiments, we introduce a suitable extension of the classical model. The model correctly describes the position of the denaturation regions, the transition boundaries, and the measured values of the DNA extension fluctuations.

Photon correlation spectroscopy with incoherent light.

Photon correlation spectroscopy (PCS) is based on measuring the temporal correlation of the light intensity scattered by the investigated sample. A typical setup requires a temporally coherent light source. Here, we show that a short-coherence light source can be used as well, provided that its coherence properties are suitably modified. This results in a "skewed-coherence" light beam allowing that restores the coherence requirements. This approach overcomes the usual need for beam filtering, which would reduce the total brightness of the beam.

[Digital clubbing]. / L'hippocratisme digital.

Digital clubbing is a clinical sign known for centuries. Though, no theory could yet explain this entire phenomenon, neither in its primary nor its secondary form. This article reviews the actual hypotheses bringing a plausible explanation, regarding the most current pathologies associated with digital clubbing. A focus is made on pulmonary and digestive pathologies, and particularly cirrhosis. The knowledge of the mechanisms underlying finger clubbing might lead, in the future, to an effective treatment.

Cooperative effects on the compaction of DNA fragments by the nucleoid protein H-NS and the crowding agent PEG probed by Magnetic Tweezers.

BACKGROUND: DNA bridging promoted by the H-NS protein, combined with the compaction induced by cellular crowding, plays a major role in the structuring of the E. coli genome. However, only few studies consider the effects of the physical interplay of these two factors in a controlled environment. METHODS: We apply a single molecule technique (Magnetic Tweezers) to study the nanomechanics of compaction and folding kinetics of a 6 kb DNA fragment, induced by H-NS bridging and/or PEG crowding. RESULTS: In the presence of H-NS alone, the DNA shows a step-wise collapse driven by the formation of multiple bridges, and little variations in the H-NS concentration-dependent unfolding force. Conversely, the DNA collapse force observed with PEG was highly dependent on the volume fraction of the crowding agent. The two limit cases were interpreted considering the models of loop formation in a pulled chain and pulling of an equilibrium globule respectively. CONCLUSIONS: We observed an evident cooperative effect between H-NS activity and the depletion of forces induced by PEG. GENERAL SIGNIFICANCE: Our data suggest a double role for H-NS in enhancing compaction while forming specific loops, which could be crucial in vivo for defining specific mesoscale domains in chromosomal regions in response to environmental changes.

Nanoparticle characterization by using tilted laser microscopy: back scattering measurement in near field.

By using scattering in near field techniques, a microscope can be easily turned into a device measuring static and dynamic light scattering, very useful for the characterization of nanoparticle dispersions. Up to now, microscopy based techniques have been limited to forward scattering, up to a maximum of 30 degrees . In this paper we present a novel optical scheme that overcomes this limitation, extending the detection range to angles larger than 90 degrees (back-scattering). Our optical scheme is based on a microscope, a wide numerical aperture objective, and a laser illumination, with the collimated beam positioned at a large angle with respect to the optical axis of the objective (Tilted Laser Microscopy, TLM). We present here an extension of the theory of near field scattering, which usually applies only to paraxial scattering, to our strongly out-of-axis situation. We tested our instrument and our calculations with calibrated spherical nanoparticles of several different diameters, performing static and dynamic scattering measurements up to 110 degrees . The measured static spectra and decay times are compatible with the Mie theory and the diffusion coefficients provided by the Stokes-Einstein equation. The ability of performing backscattering measurements with this modified microscope opens the way to new applications of scattering in near field techniques to the measurement of systems with strongly angle dependent scattering.

Characterization of anisotropic nano-particles by using depolarized dynamic light scattering in the near field.

Light scattering techniques are widely used in many fields of condensed and soft matter physics. Usually these methods are based on the study of the scattered light in the far field. Recently, a new family of near field detection schemes has been developed, mainly for the study of small angle light scattering. These techniques are based on the detection of the light intensity near to the sample, where light scattered at different directions overlaps but can be distinguished by Fourier transform analysis. Here we report for the first time data obtained with a dynamic near field scattering instrument, measuring both polarized and depolarized scattered light. Advantages of this procedure over the traditional far field detection include the immunity to stray light problems and the possibility to obtain a large number of statistical samples for many different wave vectors in a single instantaneous measurement. By using the proposed technique we have measured the translational and rotational diffusion coefficients of rod-like colloidal particles. The obtained data are in very good agreement with the data acquired with a traditional light scattering apparatus.

[Should physical activity be contraindicated during pregnancy in relation to its potentially related risks?] / L'activité physique devrait-elle être contre-indiquée pendant la grossesse au regard des risques qui lui sont potentiellement associés ?

OBJECTIVE: This paper aimed at providing a critical analysis of data on the risks associated with physical activity of women during their pregnancy. METHODS: Cochrane Library, Google Scholar, PubMed (Medline) and Web of Sciences were searched using a combination of MeSH terms associated to "Physical activity", "Pregnancy" and "Adverse events" or "Risks". Only review papers published from inception of these databases to November 2016 were used in the present analysis. RESULTS: The electronic search yield a total of 104 citations. After a critical analysis of abstracts and/or full-texts, only a systematic review and cohort study on injuries related to physical activity during pregnancy appeared relevant for the current study. Data reveals a very small proportion (<1 %) of adverse events in direct link with the physical activity behavior of pregnant women. Furthermore, there would be less than one serious adverse event per 5000hours of physical activity. The only serious adverse event attributable to physical activity during pregnancy was uterine contractions. The threat of preterm labor and miscarriage risk cannot be primarily attributed to the participation of pregnant women in a physical activity, even vigorous. Moreover, no adverse effects of maternal physical activity on neonatal outcomes were found. CONCLUSION: Physical activity-related risks during pregnancy appear to be infrequent and of minor severity. Though further studies are required to better understand the risk/benefit balance of physical activity during pregnancy, current data do not support the contraindication of this behavior in pregnant women.

Indecainide for treatment of ventricular ectopic depolarizations: efficacy, pharmacokinetics, hemodynamic effects and safety.

Ten patients were treated with oral indecainide for frequent ventricular ectopic depolarizations during a short-term, dose-ranging, single blind inpatient trial followed by open label long-term therapy for 2 years. During dose ranging, patients received placebo followed by 50, 75 and 100 mg of indecainide three times daily. Eight of the 10 patients achieved greater than or equal to 80% reduction in ventricular ectopic depolarizations during inpatient therapy. Mean ventricular ectopic depolarizations decreased from 15,792/24 h to 2,357/24 h on optimal dosage (p less than 0.01). Nine patients had paired ventricular ectopic depolarizations; four of the nine had greater than or equal to 99% reduction of these beats. Among seven patients with nonsustained ventricular tachycardia, five had 100% elimination of these events with indecainide and all had greater than or equal to 90% reduction in these events. Indecainide prolonged the PR interval 44 +/- 27 ms (p less than 0.0001) and the QRS interval 11 +/- 9 ms (p less than 0.0001) from baseline without prolongation of the QTc or JTc interval. The mean trough plasma level of indecainide on optimal dosage was 409 +/- 173 ng/ml and the mean plasma elimination half-life was 10.3 +/- 2.3 h (range 7.1 to 14.2). No adverse hemodynamic effects of indecainide were detected. Side effects during short-term therapy were mild and did not require discontinuation of the drug. Efficacy was maintained for some patients during long-term therapy for 2 years, although five patients discontinued therapy because of loss of efficacy or side effects. Indecainide is a highly effective and well tolerated antiarrhythmic drug for suppression of frequent and repetitive ventricular ectopic depolarizations.

Serum potassium, calcium and magnesium after resuscitation from ventricular fibrillation: a canine study.

Serum electrolytes were measured before and sequentially for 3 hours after resuscitation from ventricular fibrillation in a canine model that was designed to approximate the human cardiac arrest and resuscitation process. Twenty anesthetized dogs were resuscitated from ventricular fibrillation; 7 required epinephrine during resuscitation and 13 did not. To control for the effects of anesthesia, 10 dogs were anesthetized and instrumented, but ventricular fibrillation was not induced. Serum potassium decreased from 3.7 +/- 0.3 mmol/liter at baseline to 3.2 +/- 0.4 mmol/liter 45 minutes after resuscitation in the experimental dogs resuscitated without epinephrine, as compared with 3.6 +/- 0.3 to 3.4 +/- 0.2 mmol/liter in control dogs (p = 0.07 versus control dogs by two-way analysis of variance) and returned toward baseline at the end of 3 hours. Serum calcium decreased from 9.6 +/- 0.6 mg/dl at baseline to 8.9 +/- 0.9 mg/dl at 5 minutes after resuscitation as compared with 9.4 +/- 0.7 to 9.5 +/- 0.7 mg/dl in control dogs (p less than 0.05 versus control dogs) and returned to baseline by 3 hours. Serum magnesium decreased from 1.5 +/- 0.1 to 1.3 +/- 0.2 mEq/dl by 3 hours in resuscitated dogs as compared with 1.6 +/- 0.2 to 1.5 +/- 0.2 mEq/dl in control dogs (p = 0.06 versus control dogs). These changes in serum potassium, calcium and magnesium were independent of the administration of epinephrine during the resuscitation process. Changes in potassium were independent of arterial pH or bicarbonate therapy. Serum glucose increased after ventricular fibrillation but not in control dogs (p less than 0.0005 versus control). No changes in other electrolytes were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative study of encainide and quinidine in the treatment of ventricular arrhythmias.

The antiarrhythmic efficacy and safety of oral encainide hydrochloride and quinidine sulfate were compared in a nine center double-blind crossover study in 187 outpatients with benign or potentially lethal ventricular arrhythmias. Patients with at least 30 premature ventricular complexes/h were randomized to receive either encainide, 25 mg four times/day, or quinidine, 200 mg four times/day, for 2 weeks. These doses were continued for another 2 weeks if a 75% or greater reduction in premature ventricular complexes was observed. If this reduction was not seen, encainide was increased to 50 mg four times/day or quinidine to 400 mg four times/day for an additional 2 weeks. Both drugs produced a statistically significant reduction in premature ventricular complex frequency compared with baseline values. Encainide produced a statistically significant greater mean reduction in total premature ventricular complexes than did quinidine during the initial dose phase and after dose adjustment. More patients required dose increases of quinidine (60%) than of encainide (51%). Early discontinuation of treatment resulting in advancement to the next study period occurred in 12 patients taking encainide and 38 patients taking quinidine (p less than 0.05). PR and QRS intervals increased significantly during encainide treatment, as did QTc and JT intervals during quinidine treatment. No adverse reactions resulted from these electrocardiographic changes. Adverse reactions were more common with quinidine than with encainide.(ABSTRACT TRUNCATED AT 250 WORDS)

The Cardiac Arrhythmia Suppression Trial: first CAST ... then CAST-II.

The Cardiac Arrhythmia Suppression Trial (CAST) was a study designed to test the hypothesis that suppression of ventricular premature complexes after a myocardial infarction would improve survival. Preliminary results showed that suppression of ventricular premature complexes with encainide and flecainide worsened survival, and the CAST continued as the CAST-II with moricizine compared with its placebo. The protocol for the CAST-II was changed to attempt to enroll patients more likely to experience serious arrhythmias. The enrollment time was narrowed to 4 to 90 days after myocardial infarction; the qualifying ejection fraction was lowered to less than or equal to 0.40; a higher dose of moricizine could be used; early titration itself was double-blind with a placebo, and the definition of disqualifying ventricular tachycardia was changed to allow patients with more serious arrhythmias to be entered into the trial. The Cardiac Arrhythmia Suppression Trial-II was subsequently terminated prematurely because 1) patients treated with moricizine had an excessive cardiac mortality rate during the 1st 2 weeks of exposure to the drug, and 2) there appeared to be little chance of showing a long-term survival benefit from treatment with moricizine. This report outlines the rationale behind the Cardiac Arrhythmia Suppression Trial and the reasons for selection of the drugs used in the CAST and CAST-II.

A cross-sectional nationwide survey on esophageal atresia and tracheoesophageal fistula.

BACKGROUND: Our study aims at disclosing epidemiology and most relevant clinical features of esophageal atresia (EA) pointing to a model of multicentre collaboration. METHODS: A detailed questionnaire was sent to all Italian Units of pediatric surgery in order to collect data of patients born with EA between January and December 2012. The results were crosschecked by matching date and place of birth of the patients with those of diagnosis-related group provided by the Italian Ministry of Health (MOH). RESULTS: A total of 146 questionnaires were returned plus a further 32 patients reported in the MOH database. Basing on a total of 178 patients with EA born in Italy in 2012, the incidence of EA was calculated in 3.33 per 10,000 live births. Antenatal diagnosis was suspected in 29.5% patients. 55.5% showed associated anomalies. The most common type of EA was Gross type C (89%). Postoperative complications occurred in 37% of type C EA and 100% of type A EA. A 9.5% mortality rate was reported. CONCLUSIONS: This is the first Italian cross-sectional nationwide survey on EA. We can now develop shared guidelines and provide more reliable prognostic expectations for our patients.

Efficacy, safety, hemodynamic effects, and pharmacokinetics of high-dose moricizine during short- and long-term therapy.

Moricizine, 15 mg/kg, was given to 10 patients with frequent ventricular ectopic depolarizations, eight of whom had previously been treated unsuccessfully with antiarrhythmic drugs. A single-blind inpatient study was followed by therapy for up to 6 months. Two patients developed aggravation of arrhythmia during inpatient therapy. Of the eight patients who completed the inpatient study, seven achieved greater than or equal to 80% suppression of total ventricular ectopic depolarizations (P less than 0.001). During inpatient therapy the mean of the individual patients' suppression of total ventricular ectopic depolarizations was 87.9%, paired ventricular beats 99.3%, nonsustained ventricular tachycardia 99.6%, and premature atrial contractions 89.0%. Suppression was maintained during long-term therapy. The PR interval increased 27% (P less than 0.001), QRS interval increased 10% (P less than 0.0001), QTc increased 1% (P not significant), and JTc decreased 2% (P not significant). Heart rate, blood pressure, and left ventricular performance at rest and exercise were unchanged by moricizine. Moricizine half-life was 9.2 +/- 3.4 hours. Plasma levels of moricizine decreased after 10 days of therapy, suggesting induction of metabolic enzyme systems.

Pharmacodynamics and side effects of flecainide acetate.

We compared side effects with flecainide trough levels and ECG intervals among 43 patients who received flecainide for up to 34 months. Flecainide plasma levels were higher when associated with cardiovascular side effects (mean 1063 ng/ml; range 296 to 2050 ng/ml) than when no side effects occurred (mean 609 ng/ml; range 89 to 1508 ng/ml; P less than 0.001). The PR interval (P less than 0.001), QRS interval (P less than 0.001), and the rate-corrected QT interval (P less than 0.001) were greater at the time of cardiovascular side effects, but the rate-corrected JT interval was not. The therapeutic-toxic window for flecainide plasma level was 381 ng/ml (at least 50% probability of efficacy) to 710 ng/ml (less than 10% probability of cardiovascular side effects). The risk of cardiovascular side effects increases at higher plasma levels of flecainide and is associated with greater increases in the PR and QRS intervals from baseline than are routinely observed during flecainide dosing.

Encainide-induced hyperglycemia.

Twenty-three patients were treated for at least one month with encainide, a new antiarrhythmic drug. No patient was treated for hyperglycemia prior to encainide therapy. During encainide administration, five episodes of marked hyperglycemia (serum glucose level greater than or equal to 200 mg/dl) developed in four patients. (One patient received encainide twice.) The mean pretreatment glucose level was 190 +/- 69 mg/dl and rose to 397 +/- 163 mg/dl after one month of encainide therapy in patients in whom hyperglycemia developed (p less than 0.025). The glucose level was 111 +/- 27 mg/dl in nonhyperglycemic patients before encainide administration and 108 +/- 22 mg/dl after one month of encainide therapy (p = NS). There was no difference in age or encainide dosage between hyperglycemic and nonhyperglycemic patients. Treatment for hyperglycemia was given during four of the five encainide treatment periods in hyperglycemic patients. Encainide was discontinued in each of the five hyperglycemic episodes; therapeutic requirements for hyperglycemia markedly decreased. Hypoglycemic reactions to insulin occurred in two patients when encainide was stopped. Thus, encainide exacerbates hyperglycemia in some patients. These patients usually have mild hyperglycemia not requiring therapy before administration of encainide but may require insulin while receiving encainide. Treatment requirements for hyperglycemia decrease following withdrawal of encainide. The mechanism of this effect and the consequences of long-term encainide therapy on glucose metabolism are unknown.

Double-blind placebo-controlled evaluation of propafenone in suppressing ventricular ectopic activity.

The effectiveness of oral propafenone in treating ventricular premature complexes (VPCs) was assessed with a single-blind dose-ranging trial followed by a double-blind, randomized, crossover comparison of propafenone and placebo. Patients subsequently were treated with propafenone for up to 24 months. During dose ranging, the average of individual percent suppressions was 83% at the largest dose (300 mg/8 hours). During the double-blind trial, the effectiveness of propafenone was confirmed, with 7 of 12 patients achieving greater than or equal to 80% reduction in VPCs (p less than 0.05 versus double-blind placebo study). Propafenone was also effective in controlling couplets and nonsustained ventricular tachycardia. Seven patients were treated with propafenone for 24 months, during which effectiveness continued, with mean suppression ranging from 67 to 79% (p less than 0.05 versus initial single-blind placebo). Propafenone prolonged PR and QRS intervals by 16 and 18%, respectively; these prolongations continued during long-term therapy. Propafenone increased serum digoxin levels in 5 of 5 patients (mean increase 83%). Cardiovascular side effects included congestive heart failure (1 patient) and conduction abnormalities (3 patients). Thus, propafenone was effective in the treatment of total and repetitive VPCs. Side effects were few, but congestive heart failure, conduction disturbances and increases in serum digoxin were observed.