Genetic regulation of enterocyte function: a quantitative in situ hybridisation study of lactase-phlorizin hydrolase and Na(+)-glucose cotransporter mRNAs in rabbit small intestine

The enterocyte undergoes sequential changes in its structure and function as it migrates rapidly from the small intestinal crypts to the villus tip. The mechanisms by which these changes are regulated "in tune" with ontogenic and dietary changes in the luminal environment are currently under investigation. This study has employed oligonucleotide probes to follow the expression of the lactase-phlorizin hydrolase (LPH) and Na(+)-glucose cotransporter (SGLT1) genes in rabbit small intestine using quantitative in situ hybridisation histochemistry. The profiles of LPH mRNA and SGLT1 mRNA accumulation along the crypt-villus axis were found to be very similar. Although mRNA was undetectable in the crypt. LPH and SGLT1 mRNA levels rose rapidly at the crypt-villus junction, reaching a maximum between 210 microns and 330 microns above this point. Further up the villus the level of mRNAs declined. SGLT1 mRNA was present in all small intestinal segments (duodenum, jejunum and ileum), whereas LPH mRNA was absent from the ileum. LPH activity rose and fell in conjunction with mRNA, but SGLT1 activity was greatest at the villus tip where mRNA levels were considerably reduced. These data have been used to discuss the genetic regulation of enterocyte differentiation and function.     

Molecular approaches to diagnosis of Chagas' disease: use of defined antigens and a target ribosomal RNA sequence

We evaluated the performance of two defined antigens in the serological diagnosis of Chagas' disease. One of them is a recombinant protein named B13 isolated from a genomic library of Trypanosoma cruzi in the expression vector lambda gtll. We show that the gene corresponding to B13 is conserved in the evolutive stages of the two "polar" strains of T. cruzi. The protein epitopes cloned in B13 are represented in 140 kDa, 116 kDa and 35 kDa polypeptides of trypomastigotes. The other antigen chosen for serodiagnosis is a lipopeptidophosphoglycan (LPPG). This glycoconjugate is also widely distributed in T. cruzi strains. The use of a rabbit serum to LPPG allowed the demonstration that this molecule bears epitopes in common to LPPG-like components and to 80-90 kDa glycoproteins of trypomastigotes. Both B13 and LPPG were evaluated in serodiagnosis by ELISA and RIA using a panel of normal human, Chagasic and Leishmaniasis sera. It was observed that B13 presents high sensitivity and specificity for Chagasic sera. For LPPG it was also concluded that this reagent discriminates between individuals infected and non-infected with T. cruzi. A heterogeneity in the level of antibodies to LPPG in Chagasic patients was detected. No correlation was found between the clinical form of Chagas' disease and the preferential reactivity to B13 or LPPG. We also report preliminary studies towards the characterization of a 100 bp sequence of the 24S alpha rRNA as a target for DNA-based detection systems for diagnosis. We show that polymerase chain reaction of total DNA of different trypanosomatids lead to the specific amplification of a 100 bp fragment only for T. cruzi. Northern blots confirmed the presence of the target region in the mature 24S alpha rRNA. Titration experiments based on the direct amplification of RNA with Taq DNA polymerase allowed the detection of 50 parasites. Studies are in progress to increase the sensitivity of the proposed system.     

Hematologic effects of benzene. Job-specific trends during the first year of employment among a cohort of benzene-exposed rubber workers

Hematologic surveillance data from 1940 to 1975 were analyzed for a benzene-exposed cohort of 459 rubber workers. The present analyses are restricted to 161 workers with "preemployment" counts done before exposure and rely on their subsequent counts from the first 12 months of employment. While blood cell counts declined approximately 1000 cells/mm3 over the first 4 months of exposure. Using repeated-measures analysis of variance, workers exposed above the median benzene exposure at the plant had significantly lower average white and red blood cell counts at each month during the first year of work when compared with workers exposed below the median. These decreased counts suggest that clinically detectable bone marrow depression accompanied the onset of work in this plant during the 1940s and support exposure assessments that favor higher benzene levels in the 1940s when compared with subsequent decades. The general utility of repeated-measures analytic techniques for medical surveillance data is also demonstrated by this analysis.     

The influence of the pore size and its distribution in a carbon paper electrode on the performance of a PEM Fuel cell

Porous conducting carbon paper with its unique combination of properties acts as the backing material of electrode in a fuel cell. It not only assists in the flow of electrons and reactant gases but also acts as an effective support for the electrolyte and the catalyst layer. The electrically conducting porous carbon paper was prepared by adopting a modified process of preparation which involves molding together several carbon fiber preforms in the form of laminates rather than a single preform. The method was found to influence the characteristics of the paper significantly and resulted in improved performance of the unit fuel cell employing the laminated paper as electrode. The I-V performance of the fuel cell using carbon paper formed by molding single ply showed a peak power density of 573 mW/cm² as compared to that of 722 mW/cm² for three ply laminates, a value very close to that of achieved by using Toray (Japan) carbon paper (782 mW/cm²) under identical operating conditions.     

Effect of base dissipation on the granular flow down an inclined plane

The effect of base dissipation on the granular flow down an inclined plane is examined by altering the coefficient of restitution between the moving and base particles in discrete element (DE) simulations. The interaction laws between two moving particles are kept fixed, and the coefficient of restitution (damping constant in the DE simulations) between the base and moving particles are altered to reduce dissipation, and inject energy from the base. The energy injection does result in an increase in the strain rate by up to an order of magnitude, and the temperature by up to two orders of magnitude at the base. However, the volume fraction, strain rate and temperature profiles in the bulk (above about 15 particle diameters from the base) are altered very little by the energy injection at the base. We also examine the variation of h _stop , the minimum height at the cessation of flow, with energy injection from the base. It is found that at a fixed angle of inclination, h _stop decreases as the energy dissipation at the base decreases.     

An automated algorithm for online detection of fragmented QRS and identification of its various morphologies

Fragmented QRS (f-QRS) has been proven to be an efficient biomarker for several diseases, including remote and acute myocardial infarction, cardiac sarcoidosis, non-ischaemic cardiomyopathy, etc. It has also been shown to have higher sensitivity and/or specificity values than the conventional markers (e.g. Q-wave, ST-elevation, etc.) which may even regress or disappear with time. Patients with such diseases have to undergo expensive and sometimes invasive tests for diagnosis. Automated detection of f-QRS followed by identification of its various morphologies in addition to the conventional ECG feature (e.g. P, QRS, T amplitude and duration, etc.) extraction will lead to a more reliable diagnosis, therapy and disease prognosis than the state-of-the-art approaches and thereby will be of significant clinical importance for both hospital-based and emerging remote health monitoring environments as well as for implanted ICD devices. An automated algorithm for detection of f-QRS from the ECG and identification of its various morphologies is proposed in this work which, to the best of our knowledge, is the first work of its kind. Using our recently proposed time–domain morphology and gradient-based ECG feature extraction algorithm, the QRS complex is extracted and discrete wavelet transform (DWT) with one level of decomposition, using the ‘Haar’ wavelet, is applied on it to detect the presence of fragmentation. Detailed DWT coefficients were observed to hypothesize the postulates of detection of all types of morphologies as reported in the literature. To model and verify the algorithm, PhysioNet''s PTB database was used. Forty patients were randomly selected from the database and their ECG were examined by two experienced cardiologists and the results were compared with those obtained from the algorithm. Out of 40 patients, 31 were considered appropriate for comparison by two cardiologists, and it is shown that 334 out of 372 (89.8%) leads from the chosen 31 patients complied favourably with our proposed algorithm. The sensitivity and specificity values obtained for the detection of f-QRS were 0.897 and 0.899, respectively. Automation will speed up the detection of fragmentation, reducing the human error involved and will allow it to be implemented for hospital-based remote monitoring and ICD devices.     

Defect profiling in organic semiconductor multilayers

Defect depth profile study has been carried out in organic semiconductor (OSC) multilayers to characterize the buried interfaces and layers using beam based positron annihilation spectroscopy. The bilayer and trilayer heterostructures (p–n, p–p and n–p–n) comprise of organic–organic and organic–inorganic (substrate) interfaces. Our study reveals the presence of defects at the interfaces whose concentration is seen to vary with the layer thickness. The S–W correlation has been used to examine the effect of organic materials as well as thickness of the layers on the defect microstructure in multilayers. The nature and type of defects in p–p bilayer are seen to be different as compared to p–n and n–p–n multilayers. Positron mobility in OSC layers has been calculated from the fitted diffusion length which is seen to be of the same order as the effective mobility of charge carrier obtained from the measured current density–voltage (J–V) characteristics. The role of structural defects and the intrinsic electric field at the interfaces on positron systematics is also examined. Positron diffusion modeling together with experimental data suggests that the defect at the interfaces has a stronger influence on the positron systematics than the intrinsic electric field across organic–organic interfaces.     

Growth of InAs Quantum Dots on Germanium Substrate Using Metal Organic Chemical Vapor Deposition Technique

Self-assembled InAs quantum dots (QDs) were grown on germanium substrates by metal organic chemical vapor deposition technique. Effects of growth temperature and InAs coverage on the size, density, and height of quantum dots were investigated. Growth temperature was varied from 400 to 450 °C and InAs coverage was varied between 1.40 and 2.35 monolayers (MLs). The surface morphology and structural characteristics of the quantum dots analyzed by atomic force microscope revealed that the density of the InAs quantum dots first increased and then decreased with the amount of InAs coverage; whereas density decreased with increase in growth temperature. It was observed that the size and height of InAs quantum dots increased with increase in both temperature and InAs coverage. The density of QDs was effectively controlled by growth temperature and InAs coverage on GaAs buffer layer.