Neuropsychopathology in 7 Patients with the 22q13 Deletion Syndrome: Presence of Bipolar Disorder and Progressive Loss of Skills.

The 22q13 deletion syndrome is characterised by intellectual disability (ID), delayed or absent speech, autistic-like behaviour and minor, nonspecific dysmorphic features. The deletion of the SHANK3 gene is thought to be responsible for these features. In this study, the clinical data of 7 patients with the 22q13 deletion syndrome are presented, obtained by clinical genetic examination, direct behavioural observation and by interview of family members and/or caregivers, complemented by behavioural questionnaires. The specific focus was on behaviour, psychopathology and the level of functioning during life course in order to determine common features that might contribute to the delineation of the syndrome. Major findings were a high incidence of psychiatric disorders, more in particular bipolar disorder (BPD) and attention deficit hyperactivity disorder (ADHD), and a sudden deterioration after acute events, in addition to a progressive loss of skills over years. Therefore, a deletion of SHANK3 may result in a dysfunctional nervous system, more susceptible to developmental problems and psychiatric disorders on the one hand, less able to recuperate after psychiatric and somatic events, and more vulnerable to degeneration at long term on the other hand. These results are exploratory and need to be confirmed in a larger sample.

Titanium modified with layer-by-layer sol-gel tantalum oxide and an organodiphosphonic acid: a coating for hydroxyapatite growth.

Titanium and its alloys are widely used in surgical implants due to their appropriate properties like corrosion resistance, biocompatibility, and load bearing. Unfortunately when metals are used for orthopedic and dental implants there is the possibility of loosening over a long period of time. Surface modification is a good way to counter this problem. A thin tantalum oxide layer obtained by layer-by-layer (LBL) sol-gel deposition on top of a titanium surface is expected to improve biocorrosion resistance in the body fluid, biocompatibility, and radio-opacity. This elaboration step is followed by a modification of the tantalum oxide surface with an organodiphosphonic acid self-assembled monolayer, capable of chemically binding to the oxide surface, and also improving hydroxyapatite growth. The different steps of this proposed process are characterized by surfaces techniques like contact angle, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM).

Morphology of polystyrene films deposited by RF plasma.

A new plasma reactor, set up with a large planar inductively coupled source, is used for the first time to deposit a polymer coating (pPS) from a styrene monomer. This work is devoted to the relationship between external plasma parameters and substrate topography, and pPS coating morphology, which is investigated by scanning electron microscopy and atomic force microscopy. Stainless steel, gold and glass surfaces are used as substrates. It is clearly demonstrated that the film morphology can be controlled by adjustment of RF input power, pressure. The analysis performed further reveals that the pPS film's characteristics strongly depend on the substrate topography and its electrical potential during the discharge. Finally, the plasma duration also strongly influences the morphology of the films. The morphologies obtained include smooth films without any specific feature, worm-like structures, particles (nanometer- and micrometer-sized) associated along preferential directions and randomly distributed particles (micrometer-sized). The intrinsic topography of the substrate influences the film structure in the case of thin films (thickness lower than about 100 nm). Polymerization is suggested to take place at the surface in contact with the discharge rather than in the gas phase. Nucleation and growth start preferentially on substrate defects such as polishing scratches.

Influence of DNA condensation state on transfection efficiency in DNA/polymer complexes: an AFM and DLS comparative study.

Atomic force microscopy (AFM) is used to describe the formation process of polymer/DNA complexes. Two main objectives of this research are presented. The first one is to apply AFM as an effective tool to analyse DNA molecules and different polycation/DNA complexes in order to evaluate their degree of condensation (size and shape). The other one is to search for a relationship between the condensation state of DNA and its transfection efficiency. In this study, linear methacrylate based polymers and globular SuperFect polymers are used in order to induce DNA condensation. Ternary complexes, composed of methacrylate based polymers and polyethylene glycol (PEG)-based copolymers, are also investigated. AFM allows us to confirm good condensation conditions and relate them (or not) to transfection efficiencies. These AFM results (obtained after drying in air) are compared with measurements deduced from Dynamic Light Scattering (DLS) experiments performed in water. This comparison allowed us to identify the structural modifications resulting from deposition on the mica surface.

Electronic and molecular properties of an adsorbed protein monolayer probed by two-color sum-frequency generation spectroscopy.

Two-color sum-frequency generation spectroscopy (2C-SFG) is used to probe the molecular and electronic properties of an adsorbed layer of the green fluorescent protein mutant 2 (GFPmut2) on a platinum (111) substrate. First, the spectroscopic measurements, performed under different polarization combinations, and atomic force microscopy (AFM) show that the GFPmut2 proteins form a fairly ordered monolayer on the platinum surface. Next, the nonlinear spectroscopic data provide evidence of particular coupling phenomena between the GFPmut2 vibrational and electronic properties. This is revealed by the occurrence of two doubly resonant sum-frequency generation processes for molecules having both their Raman and infrared transition moments in a direction perpendicular to the sample plane. Finally, our 2C-SFG analysis reveals two electronic transitions corresponding to the absorption and fluorescence energy levels which are related to two different GFPmut2 conformations: the B (anionic) and I forms, respectively. Their observation and wavelength positions attest the keeping of the GFPmut2 electronic properties upon adsorption on the metallic surface.

Bayesian forecasting of oral cyclosporin pharmacokinetics in stable lung transplant recipients with and without cystic fibrosis.

The aims of the current study were (1) to study Neoral pharmacokinetics (PK) in stable lung recipients with or without cystic fibrosis (CF), (2) to compare Neoral PK between these two groups, and (3) to design Bayesian estimators for PK forecasting and dose adjustment in these patients using a limited number of blood samples. The individual PK of 19 adult lung transplant recipients, 9 subjects with CF and 10 subjects without CF, were retrospectively studied. Three profiles obtained within 5 days were available for each patient. A PK model combining a gamma distribution to describe the absorption profile and a two-compartment model were applied. Different exposure indices were estimated using nonlinear regression and Bayesian estimation. The PK model developed reliably described the individual PK of Neoral in lung transplant patients with and without CF, and the values of the first and second half-lives were different in these two populations (lambda(1) = 4.14 +/- 3.01 vs. 2.16 +/- 1.75 h(-1); P < 0.01; lambda(2) = 0.36 +/- 0.11 vs. 0.49 +/- 0.12 h(-1); P < 0.01), while the mean absorption time and standard deviation of absorption time tended to be less in patients with cystic fibrosis (P < 0.1). Also, the patients with CF required higher doses than those without CF to achieve similar drug exposure. Consequently, population modeling was performed in CF and non-CF patients separately. Bayesian estimation allowed accurate prediction of AUC(0-12), AUC(0-4), C(max), and T(max) using three blood samples collected at T0h, T1h, and T3h in both groups. This study demonstrated the applicability and good performance of the PK model previously developed for oral cyclosporin and of the MAP Bayesian estimation of cyclosporin systemic exposure in CF and non-CF patients. Moreover, it is the first to propose a monitoring tool specifically designed for cyclosporin monitoring in patients with CF.

[The medical chemistry department]. / Le Service de Chimie Médicale.

The laboratory of clinical chemistry performs more than 300 different tests in biochemistry, hormone and tumor markers analysis, therapeutic drug monitoring and toxicology. For the most basic tests it has followed the trend of clinical chemistry towards automation and since 2001 the heart of the laboratory is a modular automated system (MODULAR) including a preanalytical platform, unique in Belgium. For more sophisticated tests, the most recent techniques have been implemented, in particular capillary electrophoresis and ICP-MS ("inductively coupled plasma-mass spectrometry). Since 1994, the laboratory has become a reference center in the field of erythrocyte hereditary diseases, combining screening, diagnosis and research. The other research themes are the physiopathology of first trimester pregnancy and the P2Y receptors of extracellular nucleotides.

Analyzer transfer of a broad range high-sensitivity C-reactive protein immunoassay.

C-reactive protein (CRP) is measured in two main clinical situations: inflammation, where levels in the range of 5-300 mg/L are expected, and, more recently, assessment of the cardiovascular risk, where concentrations between 0.1 and 10 mg/L shoud be determined. Few commercially available methods display a measuring range covering both zones and laboratories are compelled to use two different assay protocols or even two different methods. The aim of the study was to adapt the Roche C-Reactive Protein (Latex) kit, initially developed for the Roche Cobas Integra analyzers, to the Hitachi Modular P800 analyzer in order to obtain on this instrument a broad range assay for CRP measurement. The method was successfully adapted and validated against a high-sensitivity and a traditional assay. The resulting method correlates well with the other two and displays a measuring range of 0.10-171 mg/L with an imprecision lower than 5.5%. This assay could be particularly practical in the routine clinical laboratory, being suitable for every use of CRP measurements.

Measurement of soluble transferrin receptor by immunoturbidimetry and immunonephelometry.

OBJECTIVE: To evaluate and compare two new commercially available immunoturbidimetric and immunonephelometric assays for measuring soluble transferrin receptor (sTfR) in serum. To adapt the immunonephelometric assay to an automated chemistry analyzer. DESIGN AND METHODS: Total imprecisions and detection limits were calculated and compared. Fifty-six samples were used for methods comparison. The nephelometric assay was adapted to the Hitachi 911 analyzer. RESULTS: Both methods displayed acceptable imprecisions and detection limits. Their correlation was good but a proportional bias was observed. The adaptation of the nephelometric assay to the Hitachi 911 was successful. CONCLUSIONS: These two new immunoassays are analytically acceptable and more practicable than previously developed methods. The differences observed between both methods underscores the need for standardization. The nephelometric assay was easily adaptable to an automated chemistry analyzer.

Angelman syndrome in three adult patients with atypical presentation and severe neurological complications.

Angelman syndrome (AS) is a distinct neurogenetic disorder and the phenotype is well known in childhood and adolescence. However, with advancing age the clinical and behavioral phenotype changes. In adulthood, the phenotype can be rather aspecific. We report on AS in 3 severely to profoundly mentally retarded patients, who developed severe neurologic complications of severe tremor, spasticity and coordination problems, resulting into severe loss of function. They presented atypical craniofacial features, short stature, epileptic seizures, microcephaly, brachytelephalangy and absent speech. Two patients presented at an older age a change in day-night rhythm. Based on this experience, we conclude that all severely to profoundly mentally retarded patients with atypical phenotype, spasticity, absent speech, epileptic seizures and changed day-night rhythm are candidates for further cytogenetic and molecular investigation for AS. Clinical photographs of the patient at a younger age can be helpful. The presence of the typical EEG pattern with frontal triphasic delta waves may direct to the diagnosis of AS.

Maternal tobacco exposure and cotinine levels in fetal fluids in the first half of pregnancy.

OBJECTIVE: To evaluate the distribution of cotinine in fetal fluids and serum during the first half of pregnancy, and compare the fetal and maternal cotinine levels in passive and active smokers. METHODS: Maternal smoking status was determined by questionnaire in 85 pregnant women requesting abortion for psychosocial reasons between 7 and 17 weeks' gestation. Coelomic and amniotic fluid samples were collected between 7 and 11 weeks and fetal blood and amniotic fluid between 11 and 17 weeks. Cotinine levels were measured by radioimmunoassay. RESULTS: Women classified themselves as nonsmokers in 40 cases, passive smokers in 19 cases, and voluntary smokers in 26 cases. Five nonsmokers, 16 passive smokers, and all smokers had cotinine levels above the detection limit of the assay. Cotinine was invariably found in coelomic, amniotic, and fetal serum when maternal serum and urine cotinine levels exceeded 25 and 250 ng/mL, respectively. Higher cotinine levels were found in fetal fluids and serum than in maternal serum. Positive linear correlations were found between maternal urine and amniotic fluid cotinine concentrations (r = .75), between maternal urine cotinine concentration and number of cigarettes smoked per day (r = .66), and between maternal and fetal serum cotinine concentrations (r = .97). CONCLUSION: Cotinine accumulates in the fetal compartments as early as 7 weeks' gestation in both active and passive smokers. Women should be advised to give up smoking from conception and avoid environmental tobacco smoke exposure.

Placental transfer of fentanyl in early human pregnancy.

To investigate the transfer of fentanyl across the early human placenta, we have collected samples of maternal blood and fetal fluids and/or blood, simultaneously, between 5 and 22 min following an intravenous bolus of fentanyl (1.5 microg/kg) to the mother. The pregnancies were between 6 and 16 weeks of gestation and scheduled for elective termination of pregnancy under general anaesthesia. Total fentanyl concentration was determined by radioimmunoassay in 11 pairs of first trimester maternal serum and fetal coelomic fluid samples, 14 pairs of maternal serum and amniotic fluid samples, seven series of first trimester maternal serum and coelomic and amniotic fluid samples, and 10 series of early second trimester maternal and fetal sera and amniotic fluid samples. Fentanyl was not detected in coelomic fluid samples at any gestational age and in amniotic fluid samples collected after 12 weeks of gestation. Measurable concentrations of fentanyl were found in maternal serum collected within 15 min after the initial bolus and in fetal serum collected between 10 and 12 min later. These findings indicate that fentanyl is transferred across the early placenta into the amniotic cavity and fetal blood circulation but not into the exocoelomic cavity. The distribution of this molecule inside the early gestational sac is probably influenced by the increased binding by maternal and fetal sera, its short half-life of distribution and the specific biology of the fetal fluid formation and composition.

Whole blood cyclosporin monitoring in liver and heart transplant patients: evaluation of the specificity of a fluorescence polarization immunoassay and an enzyme-multiplied immunoassay technique.

The specificity of two cyclosporin immunoassays were evaluated. Eleven patients were followed for the first four weeks after heart (n = 3) or liver (n = 8) transplantation. Cyclosporin A (CsA) monitoring was performed concomitantly by a monoclonal fluorescence polarization immunoassay (mFPIA) and enzyme-multiplied immunoassay technique (EMIT) during this period. For several patients, cyclosporin monitoring was also performed by high performance liquid chromatography (HPLC) or by polyclonal fluorescence polarization immunoassay (pFPIA). Liver function was assessed by follow-up of plasma total bilirubin, gamma-glutamyl transferase and alkaline phosphatase and renal function by plasma creatinine. All the patients presented episodes of impaired liver function. Higher CsA levels were found using mFPIA measurements as compared to the EMIT measurements (ratio mFPIA:EMIT (medium range) = 1.4 (1.0-2.3)). A higher degree of cross-reactivity of the antibody used in the mFPIA as compared to the EMIT was demonstrated by specific measurements of CsA and its primary metabolite, AM1, by HPLC.

Prognostic evaluation of patients with parenchymal cirrhosis. Proposal of a new simple score.

BACKGROUND/AIMS: The current shortage of organs makes it desirable to establish the prognosis of patients with cirrhosis in order to assess priority for liver transplantation. METHODS: We compared the utility of two exogenous tests (aminopyrine breath test and lidocaine metabolization test), two clinical parameters (encephalopathy, ascites), 18 endogenous tests and five scores (Pugh, Merkel, Orrego, Adler, Pignon) for predicting 1-year mortality in patients with parenchymal cirrhosis. Retrospective (n=49 out of 63 patients) and prospective (n=38 out of 46 patients) series were included. Univariate, multivariate, receiver operator curves and survival curves were employed. RESULTS: We found that endogenous tests were more discriminant than exogenous tests. The best parameters of the univariate analysis (encephalopathy, bilirubin, alkaline phosphatase, cholinesterase and bile acids) and their 25th and 75th percentiles were included in an additive new score which turned out to be superior to the five other scores. Prospectively, the sensitivity of our new score compared to the Pugh score was 82% versus 95% (NS) and the specificity was 89% versus 56% (p<0.01). CONCLUSIONS: Our new simple score appears to be very powerful for predicting prognosis at 1 year for patients with cirrhosis and should be evaluated in other centers.

Systemic and intrastriatal theophylline have opposite effects on dopamine and dopamine metabolites measured by intrastriatal microdialysis in the rat.

Using a model of intrastriatal microdialysis, we studied the effect of theophylline, an A1 and A2A adenosine receptor antagonist on striatal dopamine (DA) and DA metabolites. Systemic administration of theophylline (10 and 50 mg/kg) significantly reduced striatal extracellular (EC) levels of DA and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and 4-hydroxy-3-methoxy-phenylacetic acid (HVA). Intrastriatal administration of theophylline (10(-2) M) significantly increased DA and its metabolites (DA1 + 120%; DOPAC, +28%; HVA, +30%). Contradictory effects of systemic and intrastriatal theophylline point to theophylline interactions with different receptors possibly at different locations.

Trypsin activity. A new marker of acute alcoholic pancreatitis.

A normal serum amylase level is found in up to 32% of patients with acute alcoholic pancreatitis. This underlines the need for more sensitive diagnostic tests in this frequent cause of pancreatitis. Animal and human studies have shown that chronic alcohol consumption leads to important modifications in trypsinogen metabolism. The present work has prospectively analyzed admission serum trypsin activity with a new biochemical test and usual markers such as amylase, lipase, and immunoreactive trypsin in 32 attacks of acute pancreatitis. Seventeen were due to alcohol and 15 to other causes, including 11 with gallstone pancreatitis. High trypsin activity (median: 235 units/liter; range: 165-853) was found in all patients with acute alcoholic pancreatitis even when the amylase level was normal on admission (3/17: 18%). Trypsin activity did not differ between nonalcoholic pancreatitis (N = 15): 84 units/liter (42-98), alcoholic controls (N = 15): 77 units/liter (40-122), and healthy controls (N = 62): 81 units/liter (15-143). The difference was not related to the severity of disease or circulating alpha 2-macroglobulin, alpha 1-protease inhibitor, or immunoreactive trypsinogen levels. Lipase/amylase ratio was less discriminant than trypsin activity between alcoholic and nonalcoholic diseases. We conclude that serum trypsin activity seems specific to acute alcoholic pancreatitis and should be included in new prospective studies assessing biochemical testing of alcohol-related pancreatic diseases.

Distribution of iron and iron-binding proteins in first-trimester human pregnancies.

OBJECTIVE: To investigate the iron distribution between the maternal and embryonic compartments in the first trimester of pregnancy. METHODS: Coelomic and amniotic fluids (AF) and maternal serum were collected from 36 apparently normal pregnancies at 7-13 weeks of gestation. Iron, transferrin, ferritin, and lactoferrin were measured in all samples. Iron concentrations were also measured in placental villi, liver, gut, and brain samples collected from two embryos. RESULTS: Significantly (median value) lower iron and transferrin levels and higher levels of ferritin were found in the coelomic fluid (iron 4.8 mumol/L; transferrin 0.22 g/L) than in maternal serum (iron 21 mumol/L; transferrin 2.5 g/L). The AF contained significantly lower levels of iron and ferritin (iron less than 1.8 mumol/L; ferritin 2.0 micrograms/L) than both coelomic fluid (iron 4.8 mumol/L; ferritin 287 micrograms/L) and maternal serum (iron 21 mumol/L; ferritin 49 micrograms/L). Transferrin was undetectable (less than 0.08 g/L) in AF samples, and lactoferrin was undetectable (less than 2 micrograms/mL) in both embryonic fluids. The iron concentration in the coelomic fluid increased significantly (P < .001) with advancing gestation (iron at 7-9 weeks 3.8 mumol/L; 9.1-11 weeks 5.9 mumol/L). There was a nonsignificant correlation between coelomic fluid and maternal serum iron and iron-binding protein levels. The highest iron levels were found in the liver (52 mmol/kg dry weight) and brain (49 mmol/kg dry weight) tissues. CONCLUSIONS: The distribution of iron and iron-binding proteins between the maternal and embryo-placental compartments in the first trimester is comparable to that found later in gestation, suggesting that placental iron transfer may occur as early as tertiary villi are formed. The exocoelomic fluid is probably the main iron reservoir in early pregnancy, and the secondary yolk sac is probably the principal route of entry of iron to the embryo.

Ectomorphic habitus, severe mental retardation and characteristic face: a new MCA/MR syndrome?

Here we report on 2 mentally retarded sisters with clinical signs and symptoms not seen in a previously delineated MCA/MR syndrome, i.e., normal pre- and perinatal history, severe mental retardation with severe delay in psychomotor development and without development of primary motor abilities and speech, characteristic face with maxillary hypoplasia, large mouth with down-turned corners, short philtrum and everted lower lip, associated with a remarkable ectomorphic habitus.