Hepatocytes cocultured with Sertoli cells in bioreactor favors Sertoli barrier tightness in rat.

The lack of a reliable in vitro system to assess reprotoxicity is an emerging problem in the context of European law for Registration, Evaluation, Authorization and Restriction of Chemicals (REACH, 2007), as it requires a reduction in animal utilization for testing. Furthermore, in vitro reprotoxicological tests would be more relevant and greatly improved by integrating both hepatic metabolism and the blood-testis barrier. Here, we took advantage of an integrated insert in a dynamic microfluidic platform (IIDMP) to co-cultivate hepatocytes in biochip and Sertoli cells in the bicameral chamber. This microfluidic tool has been previously demonstrated to be helpful in cell function and/or quality improvement. We demonstrate that permeability of the Sertoli barrier is reduced by dynamic coculture in our system. Exometabolomics analysis reveals that interactions between hepatocytes and Sertoli cells may have been mediated by the polyamines increase and/or mid-chain fatty acid decrease in the circulating medium. These metabolic changes may be involved in permeability reduction by contributing to modifying junction protein quantity and localization. The present study gives an example of IIDMP as an in vitro partitioning/transport model for cell culture and toxicological testing. Further, based on both our previous results using an intestinal-hepatic cell coculture and the present study, IIDMP seems to be well-suited for (i) assessing the dose-response effect of chemicals within the rodent or human male reproductive tract, and (ii) improving the quality of reprotoxicological assays by including hepatic metabolism. Copyright © 2016 John Wiley & Sons, Ltd.

Fluorescence in situ hybridization of Microcystis strains producing microcystin using specific mRNA probes.

Cyanobacteria are ubiquitous micro-organisms that can produce toxic compounds, the cyanotoxins. The monitoring of such producers in the environment is of prime importance for human health. An attractive technology for such monitoring is fluorescence in situ hybridization (FISH), which allows the detection and enumeration of environmental micro-organisms. We present here the application of tyramide signal amplification fluorescence in situ hybridization (TSA-FISH) to the detection of microcystin-producing Microcystis strains. We used a 16S rRNA-specific probe, MICR3, to specifically label and observe by epifluorescence microscopy Microcystis aeruginosa strains. Using confocal laser scanning microscopy and a specific probe, MCYA, targeting the mcyA mRNA we have labelled M. aeruginosa PCC 7806, which produces microcystins. Microcystis aeruginosa PCC 7005 which does not produce microcystins is not labelled by this probe. Furthermore, we show here that this specific mRNA labelling in M. aeruginosa PCC 7806 is enhanced in cells illuminated for 1 h just after a dark period of cultivation of 24 h, conditions in which the mcyA gene is up regulated. The data presented here might be applicable to the monitoring of toxic Microcystis strains in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Cyanobacteria producing toxic compounds (cyanotoxins) are present in the environment and in water bodies. Their presence poses a threat on human and animal health. It is thus important to detect, identify and enumerate these toxic Cyanobacteria. Using tyramide signal amplification fluorescence in situ hybridization (TSA-FISH) and specific probes, with confocal laser scanning microscopy, we have specifically detected Microcystis strains producing microcystin toxins. The data presented here might be applied to the monitoring of water bodies at early stages and all along the formation of Microcystis blooms.

Combining synchrotron radiation techniques for the analysis of gold coins from the Roman Empire.

Four gold coins minted in the V century have been studied with non-destructive synchrotron radiation techniques, namely X-Ray Fluorescence (XRF) and X-ray Absorption Near Edge Spectroscopy (XANES). XRF data analyzed coupling standard and statistical methods were used to distinguish the composition of the alloy constituting the coins from that of successive deposits processes. Our analysis presents a quantification of the trace elements present in the metallic alloy providing interesting details for historical insight. Furthermore, on the basis of the XRF maps, some regions of interest were selected for XANES at the K-edge of Fe. Our analysis of the Fe spectra points out two main phases which can be related to Fe oxides naturally present in soil. From the relative abundance of these oxides, information on the site where the coins were found can be obtained, providing additional information on their fate across the centuries.

How the anisotropy of surface oxide formation influences the transient activity of a surface reaction.

Scanning photoelectron microscopy (SPEM) and photoemission electron microscopy (PEEM) allow local surface analysis and visualising ongoing reactions on a µm-scale. These two spatio-temporal imaging methods are applied to polycrystalline Rh, representing a library of well-defined high-Miller-index surface structures. The combination of these techniques enables revealing the anisotropy of surface oxidation, as well as its effect on catalytic hydrogen oxidation. In the present work we observe, using locally-resolved SPEM, structure-sensitive surface oxide formation, which is summarised in an oxidation map and quantitatively explained by the novel step density (SDP) and step edge (SEP) parameters. In situ PEEM imaging of ongoing H2 oxidation allows a direct comparison of the local reactivity of metallic and oxidised Rh surfaces for the very same different stepped surface structures, demonstrating the effect of Rh surface oxides. Employing the velocity of propagating reaction fronts as indicator of surface reactivity, we observe a high transient activity of Rh surface oxide in H2 oxidation. The corresponding velocity map reveals the structure-dependence of such activity, representing a direct imaging of a structure-activity relation for plenty of well-defined surface structures within one sample.

Ab initio study of the uranyl oxide hydrates: a proton transfer mediated by water.

We present a first-principles study of the UO(3)·n(H(2)O) uranyl oxide hydrates, namely, schoepite (n = 2.25), metaschoepite (n = 2) and dehydrated metaschoepite (n = 1.75), which appear as the alteration U(VI) products of aqueous corrosion of nuclear fuel. For these compounds, the calculated enthalpy of formation is in good agreement with calorimetry and solubility measurements. We discuss the key electronic state factors behind the phase stability of uranyl oxide hydrates. An unexplored proton-transfer mechanism, which produces the H(3)O hydronium ions in UO(3)·nH(2)O, has been studied using ab initio molecular dynamics simulations at room temperature. For the hydronium ion, a very short lifetime of around 20 fs has been suggested.

Switching from 'unipolar' to bipolar II. An 11-year prospective study of clinical and temperamental predictors in 559 patients.

BACKGROUND: Given the therapeutic and prognostic importance of the unipolar-bipolar dichotomy, predicting which patients will become bipolar subsequent to index diagnosis of major depressive disorder (MDD) is of paramount clinical significance. We sought to characterize the profile of patients with MDD who would convert to the more subtle bipolar subtype (known as BPII) on the basis of clinical and personality variables obtained during MDD episodes. METHODS: A total of 559 patients, comprehensively evaluated with the Schedule of Affective Disorders and Schizophrenia and "unipolar" MDD at entry, were administered 17 self-report personality measures. Hypomanic and manic episodes were systematically recorded over a prospective observation period of up to 11 years. We compared 48 converters to BPII (8.6%) with 22 converters to bipolar I (BPI) (3.9%) and the remaining larger group of unipolar patients. RESULTS: Except for greater acuteness, severity, and psychotic symptomatology, BPI converters were essentially similar to MDD nonconverters. By contrast, BPII converters were robustly distinguished from those with MDD who remained unipolar on the basis of self-report measures along the newly derived factors of Mood Lability, Energy-Activity, and Daydreaming. This profile was associated with early age at onset of MDD and pleomorphic psychopathology beyond the usual affective realm, high rates of substance abuse, as well as educational, marital, and occupational disruption and minor antisocial acts prior to discrete hypomanic episodes. Overall, BPII switchers had a more protracted and tempestuous course with shorter well intervals. "Habitual self" descriptions of temperamental instability during MDD episodes provided useful clinical information for predicting which depressed patients will switch to BPII, attaining a sensitivity of 91% for all three factors combined (23 items); Mood Lability alone (nine items) was the most specific predictor (86%), though of lower sensitivity (42%). CONCLUSIONS: The BPII subtype is best understood by such lability intruding into, and possibly its accentuation during, depressive episodes, thereby creating an intimate interweaving of trait and state. Clinicians must note that the foregoing temperamental profile appears more fundamental in defining the affective dysregulation of the BPII subtype than hypomanic episodes emphasized in DSM-IV.

Psychosocial disability during the long-term course of unipolar major depressive disorder.

BACKGROUND: The goal of this study was to investigate psychosocial disability in relation to depressive symptom severity during the long-term course of unipolar major depressive disorder (MDD). METHODS: Monthly ratings of impairment in major life functions and social relationships were obtained during an average of 10 years' systematic follow-up of 371 patients with unipolar MDD in the National Institute of Mental Health Collaborative Depression Study. Random regression models were used to examine variations in psychosocial functioning associated with 3 levels of depressive symptom severity and the asymptomatic status. RESULTS: A progressive gradient of psychosocial impairment was associated with a parallel gradient in the level of depressive symptom severity, which ranges from asymptomatic to subthreshold depressive symptoms to symptoms at the minor depression/dysthymia level to symptoms at the MDD level. Significant increases in disability occurred with each stepwise increment in depressive symptom severity. CONCLUSIONS: During the long-term course, disability is pervasive and chronic but disappears when patients become asymptomatic. Depressive symptoms at levels of subthreshold depressive symptoms, minor depression/ dysthymia, and MDD represent a continuum of depressive symptom severity in unipolar MDD, each level of which is associated with a significant stepwise increment in psychosocial disability.

A prospective 12-year study of subsyndromal and syndromal depressive symptoms in unipolar major depressive disorders.

BACKGROUND: Investigations of unipolar major depressive disorder (MDD) have focused primarily on major depressive episode remission/recovery and relapse/recurrence. This is the first prospective, naturalistic, long-term study of the weekly symptomatic course of MDD. METHODS: The weekly depressive symptoms of 431 patients with MDD seeking treatment at 5 academic centers were divided into 4 levels of severity: (1) depressive symptoms at the threshold for MDD; (2) depressive symptoms at the threshold for minor depressive or dysthymic disorder (MinD); (3) subsyndromal or subthreshold depressive symptoms (SSDs), below the thresholds for MinD and MDD; and (4) no depressive symptoms. The percentage of weeks at each level, number of changes in symptom level, and medication status were analyzed overall and for 3 subgroups defined by mood disorder history. RESULTS: Patients were symptomatically ill in 59% of weeks. Symptom levels changed frequently (1.8/y), and 9 of 10 patients spent weeks at 3 or 4 different levels during follow-up. The MinD (27%) and SSD (17%) symptom levels were more common than the MDD (15%) symptom level. Patients with double depression and recurrent depression had more chronic symptoms than patients with their first lifetime major depressive episode (72% and 65%, respectively, vs 46% of follow-up weeks). CONCLUSION: The long-term weekly course of unipolar MDD is dominated by prolonged symptomatic chronicity. Combined MinD and SSD level symptoms were about 3 times more common (43%) than MDD level symptoms (15%). The symptomatic course is dynamic and changeable, and MDD, MinD, and SSD symptom levels commonly alternate over time in the same patients as a symptomatic continuum of illness activity of a single clinical disease.

Analogues and derivatives of tenoxicam. 1. Synthesis and antiinflammatory activity of analogues with different residues on the ring nitrogen and the amide nitrogen.

The synthesis of tenoxicam, 4-hydroxy-2-methyl-N-2-pyridyl-2H-thieno[2,3-e]-1,2-thiazine-3-carboxami de 1,1-dioxide (1e), and of the analogues with various residues on the ring nitrogen and the amide nitrogen is described. This new class of "oxicams" has pronounced antiinflammatory and analgesic properties. The very specific structure-activity relationship of isomeric and isosteric groups at the amide nitrogen has been evaluated. The substituent in position 2 also has a great influence on the pharmacological properties. Tenoxicam is presently undergoing clinical trials.

Single-pad scheme for integrated optical fluorescence sensing.

A new scheme for integrated optical fluorescence sensing is presented. The principle is based on a planar waveguide containing multiple sensing units, each consisting of a single-pad grating coupler structure. Single-pad means that all the following functions are incorporated in one single pad: laser light input, excitation of the labeled analyte molecules, efficient collection of the emitted fluorescent light into the waveguide, background suppression, and coupling the guided wave out to the detector. The results demonstrate a high efficiency of the fluorescence light excitation and collection, as well as a good suppression of the volume background.

Major depressive disorder: a prospective study of residual subthreshold depressive symptoms as predictor of rapid relapse.

BACKGROUND: The study tested whether level of recovery from major depressive episodes (MDEs) predicts duration of recovery in unipolar major depressive disorder (MDD) patients. METHODS: MDD patients seeking treatment at five academic centers were followed naturalistically for 10 years or longer. Patients were divided on the basis of intake MDE recovery into residual depressive symptoms (SSD; N=82) and asymptomatic (N=155) recovery groups. They were compared on time to first episode relapse/recurrence, antidepressant medication, and comorbid mental disorders. Recovery level was also compared to prior history of recurrent MDEs ( > 4 lifetime episodes) as a predictor of relapse/recurrence. RESULTS: Residual SSD compared to asymptomatic recovery patients relapsed to their next MDE > 3 times faster (median=68 vs. 23 weeks) and to any depressive episode > 5 times faster (median=33 vs. 184 weeks). Residual SSD recovery status was significantly associated with early episode relapse (OR=3.65) and was stronger than history of recurrent MDEs (OR=1.64). Rapid relapse in the SSD group could not be attributed to higher comorbidity or lower antidepressant treatment. LIMITATIONS: Although inter-rater agreement on weekly depressive symptom ratings was very high (ICC > 0.88), some error may exist in assigning recovery levels. Antidepressant treatments were recorded, but were not controlled. CONCLUSIONS: MDE recovery is a powerful predictor of time to episode relapse/recurrence. Residual SSD recovery is associated with very rapid episode relapse which supports the idea that SSD is an active state of illness. Asymptomatic recovery is associated with prolonged delay in episode recurrence. These findings of this present study have important implications for the goals of treatment of MDD and for defining true MDE recovery.

Patients report positive nutrition counseling outcomes.

OBJECTIVE: Assess outcomes of patient nutrition counseling. DESIGN: A descriptive study based on the results of a telephone interview performed 2 to 8 weeks after counseling. SUBJECTS/SETTING: Subjects were 400 adult patients referred for nutrition counseling at 2 academic health centers. Of these, 274 patients received nutrition counseling during hospitalization and 126 as outpatients. STATISTICAL ANALYSIS: Descriptive statistics were used to summarize data and the Mann-Whitney U statistic and logistic regressions were used to test significant differences (P < .05) between inpatient and outpatient counseling outcomes. RESULTS: Most patients (83%) gave a partial or full description of their diet modifications and 79% had a moderate or good understanding of their diet. Most patients reported that the dietitian's advice was suited to their special needs (88%) and that they knew what to eat (83%). A majority (62%) had made dietary changes, but 17% said they had had trouble changing their diets as suggested. After talking with a dietitian, 57% felt better emotionally, 37% felt better physically, 64% felt in control of their condition, and 43% noticed improved health indicators. Initial analysis indicated that outpatients reported better outcomes than inpatients; further analysis showed that these differences could be attributed to younger ages among the outpatient sample. APPLICATIONS/CONCLUSIONS: Patient nutrition counseling has positive outcomes. Therefore, key counseling points should be introduced or reinforced in inpatient settings, in conjunction with multiple-session protocols during the pre- and/or posthospitalization continuum of care. Dietitians, managers, administrators, and credentialing agencies should work together to secure and promote the necessary physical, personnel, and financial resources to make this happen.

Direct monitoring of capillary perfusion following normovolemic hemodilution in an experimental skin-flap model.

The effects of normovolemic hemodilution on skin flap survival are studied in a recently developed skin-flap model (homozygous hairless mouse ear) in which nutritional capillary flow is monitored directly by means of intravital microscopy from the time of flap creation throughout the establishment of necrosis. Two diluting agents (dextran 60 and hydroxyethyl starch 200) are utilized. Our quantitative findings demonstrate that the amount of nonperfused tissue following flap creation in both the dextran (n = 23) and starch (n = 13) groups was significantly decreased as compared with controls (n = 19). Our qualitative observations suggest that improved hemorrheologic properties at the microcirculatory level are responsible for the observed decreased necrosis. Various mechanisms by which hemodilution may act to prevent necrosis are discussed.

Evaluation of left ventricular function after acute myocardial infarction. Two-dimensional echocardiography versus radionuclide angiography. Study of 50 patients.

Two-dimensional echocardiography and gated blood radionuclide angiography was performed in 50 patients (mean age 51.3 years; 48 men and 2 women) after acute myocardial infarction, before discharge from the hospital. The aim of this study was to compare the wall motion score, determined by two-dimensional echocardiography (2DE), with the ejection fraction obtained by radionuclide angiography (RNA). The correlation between the results obtained by 2DE and RNA was good (r = 0.75; p less than 0.0001). We conclude that 2DE is a powerful diagnostic tool for the evaluation of left ventricular function.

Pseudocolor display of regional organ blood flow determined by means of the radioactive microsphere technique.

The measurement of regional blood flow (RBF) by means of the radioactive microsphere method yields large sets of raw data. In order to display this large volume of data, we have developed a program which provides the user with a graphical illustration of RBF in different organs (e.g., heart, kidney, brain) using various projections and sections. RBF is visualized by a pseudocolor representation. This set-up enables the user to perform a semiquantitative analysis of RBF and provides a tool for the compact representation of the spatial distribution of RBF values in different parts and layers of the organs being studied. The program runs on a standard microcomputer.

[Fistula of the right coronary artery near the right ventricle. An echocardiographic study]. / Fístula de la arteria coronaria derecha hacia el ventrículo derecho. Estudio ecocardiográfico.

A case of a 42-year-old woman with a right coronary artery fistula into the right ventricle is reported; she was asymptomatic and a continuous precordial murmur was found on physical examination. We discuss the usefulness and limitation of the Echo 2-D/Doppler findings, concerning this diagnosis, which was only definitively established by angiocardiography.

[Significance of Doppler echocardiography in the diagnosis and management of children with heart disease]. / Valor da ecocardiografia-Döppler no diagnóstico e orientação da criança com doença cardíaca.

OBJECTIVE: To assess the accuracy of Doppler-echocardiography in the diagnosis and management of the child with heart disease. MATERIAL AND METHODS: Prospective echocardiographic study of 132 consecutive patients aged 1 day to 16 years old with heart disease confirmed by cardiac catheterization (n = 109) or anatomic examination (surgical = 21, necropsy = 2). RESULTS: Of the 258 cardiovascular anomalies diagnosed by cardiac catheterization and anatomical examination, 247 were correctly identified by echocardiography (sensitivity = 96%). There were 11 missed and 4 false positive diagnosis (specificity = 98%), all of them were vascular anomalies. Intracardiac anatomy and segmentary alignment were always correctly identified. The 21 cardiovascular anomalies operated without cardiac catheterization were correctly diagnosed. In 16 patients (12%) a management plan could not be established only on clinical echocardiographic grounds and, in another patient, the plan was incomplete. In the remaining 115 patients (87%) the management plan was correct and complete. CONCLUSIONS: The clinical-echocardiographic study allows: 1) an excellent anatomo-functional evaluation of the heart and great vessels, 2) surgical repair of some selected cardiovascular anomalies without previous cardiac catheterization.

[Echocardiographic evaluation of simple transposition of great vessels subjected to physiologic correction]. / Avaliação ecocardiográfica da transposição simples dos grandes vasos sujeita a correcção fisiológica.

The aim of the present study was to evaluate the cardiovascular morphology after Senning's operation for transposition of the great vessels (TGV) using Döppler-echocardiography. The study included 25 patients (pts) aged 17 to 127 months (mean = 60 +/- 27) who were evaluated 6 to 112 months (mean = 42 +/- 27) after surgery. The following parameters were analysed: right (RV) and left ventricular (LV) morphology and function; tricuspid regurgitation and its severity; caval and pulmonary venous pathways and the presence of pulmonary stenosis, its type and severity. All the 25 pts had an abnormal end-systolic interventricular septal (IVS) configuration, with left convexity in 22 (88%) and a flat septum present in 3 (12%). The LV fractional shortening (FS) ranged from 25 to 60% (mean = 41 +/- 10) and the preejection/ejection time ratio (PET/ET) from 0.18 to 0.33 (mean = 0.24 +/- 0.04). The RV mean FS ranged from 23.0 to 32.5% (mean = 26.9 +/- 3.0) and PET/ET from 0.27 to 0.46 (mean = 0.38). The RV wall motion study revealed that the IVS was hypokinetic in 3 pts (12%), akinetic in 19 (76%) and dyskinetic in 3 (12%). Three pts also had an hypokinetic antero-apical segment. TR was detected in 22 (88%) pts being trivial in 16 (64%), mild in 4 (16%) and moderate in 2 (8%). It was always possible to visualise the neo left and right atria as well as the caval-right atrial junctions. No systemic or pulmonary venous obstruction was detected. In 3 pts there was a mild subpulmonic dynamic stenosis related with septal buldging and mitral valve apposition. In conclusion, Döppler-echocardiography can provide accurate information for the follow-up of patients with TGV submitted to physiological correction and it can reduce the number and frequency of late postoperative cardiac catheterization.

[Isolated right ventricular myxoma: report of a clinical case]. / Mixoma isolado do ventrículo direito: A propósito de um caso clínico.

A clinical case of a 14 years old white female with asthenia and easy fatigue of two months duration is presented. The two-dimensional echocardiography study has identified a tumorous and homogeneous mass filling completely the right ventricle. During the cardiac surgery and with the help of extemporaneous biopsy the real nature of the tumor was confirmed-cardiac myxoma. In this report the authors discuss the incidence of cardiac tumors and particularly cardiac myxomas and point out the rarity of isolated myxomas in the right ventricle.

[Value of ventricular geometry in the evaluation of direct ventricular pressure]. / Interesse da geometria ventricular na avaliação da pressão ventricular direita.

OBJECTIVE: To determine the echocardiographic end-systolic ventricular geometry value in evaluating right ventricular systolic pressure (RVSP). MATERIAL AND METHODS: We studied prospectively 68 patients (mean age = 6.0 +/- 5.0 years), submitted to cardiac catheterization for cardiac disorders not involving left ventricular (LV) outflow tract obstruction, within 24 hours after two-dimensional echocardiographic (2D echo) examination. 2D echo evaluation of RVSP was performed using end-systolic LV transverse orthogonal diameters (TDR). The LV transverse orthogonal diameters (antero-posterior and supero-inferior) were measured on a parasternal short-axis image, at the tips of papillary muscles. 2D echo semi-quantitative evaluation of RVSP was tested correlating TDR with hemodynamic RVSP/LV systolic pressure (LVSP) ratio--group 1. We also used regression equation derived from the first 35 patients to quantify RVSP in the last 33 patients--group 2. In these cases, systolic systemic arterial pressure measured by sphygmomanometry was taken as LVSP. RESULTS: The TDR ranged from 1.0 to 2.1 (mean = 1.5 +/- 0.3) and the RVSP/LVSP ratio from 0.3 to 1.7 (mean = 0.7 +/- 0.4). All patients with RVSP/LVSP greater than or equal to 65% have TDR greater than or equal to 1.3 and when RVSP less than or equal to 35 mmHg we always obtained TDR less than or equal to 1.2. The correlation between 2D echo estimated and catheter measured RVSP shows, for group 1, r = 0.88 and y = 1.1X-0.88 and, for group 2'. r = 0.88. CONCLUSION: In the absence of LV systolic obstruction, TDR is a reliable non invasive method in evaluating the RVSP.