Interhemispheric connectivity estimated from EEG time-correlation analysis in preterm infants with normal follow-up at age of five.

Brain connectivity is associated with axonal connections between brain structures. Our goal was to quantify the interhemispheric neuronal connectivity in healthy preterm infants by automated quantitative EEG time-correlation analysis. As with advancing postmenstrual age (PMA, gestational age + postnatal age) the neuronal connectivity between left and right hemisphere increases, we expect to observe changes in EEG time-correlation with age. Thirty-six appropriate-for-gestational age preterm infants (PMA between 27-37 weeks) and normal neurodevelopmental follow-up at 5 years of age were included. Of these, 22 infants underwent 3-8 repeated EEG recordings at weekly intervals. The reduced 10-20 EEG electrode system for newborns was used with five sets of bipolar channels: central-temporal, frontal polar-temporal, frontal polar-central, temporal-occipital and central-occipital. We performed EEG time-correlation analysis between homologous channels of the brain hemispheres to identify interhemispheric similarity in EEG signal shape. For each 8 s epoch of the EEG the time-correlation values and the corresponding lag times were calculated for homologous channels on both hemispheres. In all channels, the median correlation value decreased significantly (between -40% and -60% decrease) from 27 to 37 weeks PMA, for gestational maturation. For the postnatal maturation only the central-temporal channel showed a significantly decreasing trend. In contrast, the median lag time showed no uniform change with PMA. The decreasing median correlation values in all homologous channels indicate a decrease in similarity in signal shape with advancing PMA. This finding may reflect greater functional differentiation of cortical areas in the developing preterm brain and may be explained by the increase of complex neural networks with excitatory and inhibitory circuitries.

Ion chromatography for the precise analysis of chloride and sodium in sweat for the diagnosis of cystic fibrosis.

BACKGROUND: Measurement of chloride in sweat is an essential part of the diagnostic algorithm for cystic fibrosis. The lack in sensitivity and reproducibility of current methods led us to develop an ion chromatography/high-performance liquid chromatography (IC/HPLC) method, suitable for the analysis of both chloride and sodium in small volumes of sweat. METHODS: Precision, linearity and limit of detection of an in-house developed IC/HPLC method were established. Method comparison between the newly developed IC/HPLC method and the traditional Chlorocounter was performed, and trueness was determined using Passing Bablok method comparison with external quality assurance material (Royal College of Pathologists of Australasia). RESULTS: Precision and linearity fulfill criteria as established by UK guidelines are comparable with inductively coupled plasma-mass spectrometry methods. Passing Bablok analysis demonstrated excellent correlation between IC/HPLC measurements and external quality assessment target values, for both chloride and sodium. With a limit of quantitation of 0.95 mmol/L, our method is suitable for the analysis of small amounts of sweat and can thus be used in combination with the Macroduct collection system. CONCLUSIONS: Although a chromatographic application results in a somewhat more expensive test compared to a Chlorocounter test, more accurate measurements are achieved. In addition, simultaneous measurements of sodium concentrations will result in better detection of false positives, less test repeating and thus faster and more accurate and effective diagnosis. The described IC/HPLC method, therefore, provides a precise, relatively cheap and easy-to-handle application for the analysis of both chloride and sodium in sweat.

Repeatability of the sugar-absorption test, using lactulose and mannitol, for measuring intestinal permeability for sugars.

Differential sugar-absorption tests for measuring intestinal permeability for sugars have been studied in a variety of gastrointestinal diseases. Their use in general practice has been hampered by a lack of data on reference values and repeatability of the test and the laboratory assay. In this study, we determined the reference values of the sugar-absorption test, using lactulose and mannitol as probe molecules, for children and adults. The repeatability of the test is good; linear relationship: slope, 0.825 [95% confidence interval (CI), 0.571, 1.152); intercept, 0.005 (95% CI, -0.004, 0.010). The repeatability of the laboratory assay for the sugar-absorption test is excellent; linear relationship: slope, 1.014 (95% CI, 0.870, 1.094); intercept, 0.002 (95% CI, -0.005, 0.010). The validation of the sugar-absorption test makes the test useful as a simple, noninvasive, reliable intestinal permeability test for sugars, which can be of use for clinical practice. Taking possible interfering factors into account, the sugar-absorption test can be used as a diagnostic test for enteropathy of different etiologies and evaluation of therapeutic interventions in both children and adults. Studies with the sugar-absorption test may clarify the role of intestinal permeability in the pathophysiology of a variety of gastrointestinal diseases.

Surfactant treatment effects on alveolar type II cell morphology in rabbit lungs.

The effects of exogenous surfactant administration on alveolar type II cells and the lung parenchyma were examined in adult rabbits. Natural surfactant was instilled into the left lobe of New Zealand White rabbits while the right lobe served as the control. Four hours post-instillation, the lungs were fixed by vascular perfusion. Surfactant instillation did not change alveolar type II cell size but was associated with a significant reduction in the volume fraction of lamellar bodies in type II cells (20.4% in control lobes compared with 11.9% in surfactant-treated lobes). The size distribution of lamellar body profiles was different in surfactant-treated lobes compared with control lobes, with a significant decrease in lamellar bodies > 0.8 microns in diameter and a twofold increase in lamellar bodies 0.2-0.4 microns in diameter. Composite body profile number was also increased by 87% (P < 0.05) after instillation of surfactant compared with control. Saline instillation decreased lamellar body volume fraction in type II cells but three times less than surfactant instillation. These observations are consistent with a strong stimulus for secretion of endogenous surfactant 4 h after surfactant instillation in normal adult rabbit lungs, whereas the increase in composite bodies is consistent with new lamellar body formation, probably from both de novo synthesized and exogenous natural rabbit surfactant. These observations confirm that the secretory and synthetic processes of alveolar type II cells are significantly affected by exogenous surfactant instillation.

Intracellular localization and endocytosis of brush border enzymes in the enterocyte-like cell line Caco-2.

Immunoelectron microscopy was used to localize the brush border hydrolases sucrase-isomaltase (SI) and dipeptidylpeptidase IV (DPPIV) in the human colon carcinoma cell line Caco-2. Both enzymes were detected at the microvillar membrane, in small vesicles and multivesicular bodies (MVBs), and in lysosomal bodies. In addition, DPPIV was found in the Golgi apparatus, a variety of apical vesicles and tubules, and at the basolateral membrane. To investigate whether the hydrolases present in the lysosomal bodies were endocytosed from the apical membrane, endocytic compartments were marked with the endocytic tracer cationized ferritin (CF). After internalization from the apical membrane through coated pits, CF was first recovered in apical vesicles and tubules, and larger electronlucent vesicles (early endosomes), and later accumulated in MVBs (late endosomes) and lysosomal bodies. DPPIV was localized in a subpopulation of both early and late endocytic vesicles, which contained CF after 3 and 15 min of uptake, respectively. Also, internalization of the specific antibody against DPPIV and gold labeling on cryosections showed endocytosed DPPIV in both early and late endosomes. However, unlike CF, no accumulation of DPPIV was seen in MVBs or lysosomal bodies after longer chase times. The results indicate that in Caco-2 cells the majority of brush border hydrolases present in lysosomal bodies are not endocytosed from the brush border membrane. Furthermore, the labeling patterns obtained, suggest that late endosomes may be involved in the recycling of endocytosed DPPIV to the microvilli.

Prevention of suicidal behavior in adolescents: the role and training of teachers.

In order to prevent suicide (attempts) among adolescents, special programs have been developed for secondary school teachers. Such programs inform teachers about the epidemiology, aetiology, symptomatology, and treatment possibilities of severe depression and suicidality of young people, and train them to recognize, handle, and refer such students. In this paper, the rationale is given for involving teachers in suicide prevention. A review of several training programs is presented, and some evaluative findings on the effects of training teachers about the facts of suicidal behavior are presented.