Probing heterotypic cell interactions: hepatocyte function in microfabricated co-cultures

Replacement of liver function using extracorporeal bioartificial systems has been attempted with limited success. The instability of the hepatocyte phenotype in vitro has restricted the useful lifetime of these devices. Co-cultivation of hepatocytes with mesenchymal cells is one method that has been widely utilized to stabilize the liver-specific function of isolated cells; however, co-culture has yet to be successfully incorporated in a bioreactor setting. In this study, we probed heterotypic cell interactions in co-cultures of hepatocytes and 3T3 in order to better understand the cellular microenvironment necessary to induce and stabilize liver-specific functions. Using microfabrication and conventional techniques to control the heterotypic interface, the effects of varying degrees of heterotypic interaction on tissue function (albumin and urea synthesis) were examined. Our data indicated maximal induction of liver-specific functions in cultures with maximal initial heterotypic interaction, and that induction of hepatic functions in hepatocytes was increased in the vicinity of fibroblasts as compared to hepatocytes far from the heterotypic interface. Furthermore, our data suggested that heterotypic cell contact is necessary for induction of these functions. These studies will aid in the formation of design criteria for a co-culture based bioartificial liver, as well as provide a useful tool to study the role of heterotypic and homotypic interactions in liver physiology and pathophysiology.     

Technical bases and acquisition conditions of electron-beam computed tomography

PURPOSE: In this review the technical principle and scanner characteristics of electron beam computer tomography (EBCT) are discussed. METHODS: In contrast to conventional CT, image acquisition in EBCT is achieved without mechanically moving parts. This construction allows for short acquisition times in investigating given anatomical regions (100 ms per slice) or up to 8 levels without table movement and short interscan delays (50 ms per slice). RESULTS: Depending on the nature of the investigation, the scanner can be used in the single slice, continuous volume scanning and multi slice mode. The single slice mode is used for detection and quantification of coronary calcifications and for CT angiography of the coronary vessels. Equivalent to the spiral mode in conventional CT, continuous volume scanning may be used for routine investigation of the chest and abdomen. Functional investigations of the heart and perfusion measurement of different organs can be performed in multi slice mode. Because of the geometry of the electron beam scanner, radiation exposure for certain investigations is above the exposure with conventional CT. CONCLUSION: Future developments will focus on dose efficient radiation collimation, high resolution detector systems and artefact reducing reconstruction kernels.     

Solid-phase extraction on C18 silica as a purification strategy in the solution synthesis of a 1-thio-beta-D-galactopyranoside library

A novel strategy for the purification of carbohydrate-based chemical libraries synthesized in solution was developed. Purification of reaction products was accomplished by means of solid-phase extraction enabled by protecting the 2-, 3-, 4-, and 6-hydroxyl groups of a galactose derivative as their hydrophobic O-laurates. The presence of multiple O-laurates allowed adsorption of reaction products onto C18 silica while reagents and by-products were washed away with MeOH. Products were quantitatively eluted with pentane. Purification of products using solid-phase extraction offers the combined advantages of solution synthesis (normal solution reactivity and ease of reaction monitoring) with those of solid-phase synthesis (facile product isolation permitting the use of large excesses of reagents). To demonstrate the utility of the hydrophobic recovery-procedure, tetra-O-lauroyl-beta-D-galactopyranose-1-thiol was subjected to high-yielding reactions with a panel of Michael-acceptors and an alpha-chloro ketone. The resulting ketone adducts were then either reduced to the alcohols or reductively aminated with a selection of amino acids to give 30 different 1-thio-beta-D-galactosides as mixtures of four diastereomers after removal of protecting groups. At each step, the product was separated from the reagents and their by-products by simple adsorption onto C18 silica, washing with MeOH and elution of product with pentane. After completion of the combinatorial chemistry sequence, the O-laurates were cleaved by methanolysis and the product methyl laurate in turn removed from the desired water-soluble products by C18 adsorption. Individual library members were thus conveniently produced on 10-30 mg scales at purity levels of > 90%. One of the 1-thio-beta-D-galactosides thus produced was found to be a competitive inhibitor of the beta-galactosidase from E. coli with Ki value of 1.7 microM.     

Modified intra-arterial calcium stimulation with venous sampling test for preoperative localization of insulinomas

BACKGROUND: Both genetic and environmental factors are thought to contribute to specific IgE responses, however, the relative contribution of each in the responses to individual ryegrass pollen allergens is largely unknown even though some responses to allergens have been linked to certain HLA complexes. OBJECTIVE: Using a large group of monozygotic and dizygotic twins, this study was designed to investigate the IgE binding profiles of individual ryegrass pollen (Lolium perenne) components to assess the relative contribution of genetic and environmental factors in determining IgE responses to specific allergens. METHODS: Ryegrass pollen proteins were separated by electrophoresis and immunoblotted with sera from 191 pairs of twins where at least one sibling had a SPT > 2 mm to perennial ryegrass. Concordance levels for individual ryegrass pollen components were compared between monozygotic and dizygotic twins in a subset group where both twins had SPT > 3 mm to perennial ryegrass. RESULTS: Immunoblotting revealed 23 individual IgE-binding components from ryegrass pollen. Although there was a significantly greater proportion of monozygotic twins with SPT wheals greater than 3 mm when compared with the dizygotic twins, the mean case-wise concordance for the immunoblot components was similar for both groups of twins. The mean case-wise concordance when at least four pairs of sera were involved was 44% for the MZ twins (n=11 components) and 45% for the DZ twins (n=12 components). We found no significant differences in concordance levels for any of the 23 individual components including allergens previously associated with HLA. CONCLUSION: Evidence for genetic control of allergen-specific IgE responses in a large population sample of twins to individual ryegrass allergens is limited, indicating that the IgE responses to specific ryegrass pollen allergens are determined largely by environmental factors.     

Identification of a hypoperfused segment in bull's-eye myocardial perfusion images using a feed forward neural network

Artificial neural networks are computer systems which can be trained to recognize similarities in patterns and which learn by example; one of the more straightforward types being the feed forward neural network (FFNN). We previously reported the use of FFNNs for classification of hypoperfusion patterns in bull's-eye representation of 201Tl single photon emission tomography myocardial perfusion studies and showed that, when such an image was divided into 24 segments, FFNNs could detect perfusion defects without direct comparison to a normal data base. This has been extended in this investigation to assess the ability of an FFNN, trained on data in which only a single segment was hypoperfused, to detect this abnormal segment when the hypoperfusion pattern of the other segments in the image varied. The results indicated that the network could reliably determine whether a segment was normally or under perfused, with accuracies of 99% and 100%, respectively, if all other segments were normally perfused. It could also reliably detect a normally perfused segment, even if other segments were hypoperfused, with accuracies of 95% and 98%. The network was less reliable, however, in detecting a hypoperfused segment when other segments were also hypoperfused, showing accuracies of only 74% and 88%.     

The possible significance of parallel changes in plasma lutein and retinol in Pakistani infants during the summer season

We established new cell lines from head and neck cancer patients for studies of adhesion molecules and cellular behavior in nine patients with primary or metastatic cancer treated at the Asan Medical Center. Explant cultures of fresh tumor tissue were used to develop new permanent tumor cell lines. Lines were tested for tumor formation and histology in nude mice. Flow cytometry and indirect immunofluorescence were used to assess DNA content and expression of the alpha 6, beta 4, and beta 1 integrin subunits and the intercellular adhesion molecule 1 (ICAM-1). In vitro growth patterns and adhesion to plastic were assessed using phase contrast microscopy. AMC-HN-1 to -8 were derived from patients with squamous cell carcinoma. AMC-HN-9 was from an undifferentiated carcinoma of the parotid gland. The 8 lines we tested produced nude mouse tumors that are identical to the histology of the original tumors. AMC-HN-1, -2, -5, and -9 have epithelioid or spindle cell morphology with poor cell-to-cell and cell-to-substrate adhesiveness. AMC-HN-3, -4, -7, and -8 grow as adherent epithelioid monolayers. AMC-HN-6 exhibits multilayer stratification. Four lines are near diploid, 4 are hyperdiploid and 1 is hypodiploid. Only three express ICAM-1. All lines express the alpha 6, beta 4, and beta 1 integrin subunits but to different extent. Four, AMC-HN-1, -2, -5, and -6, express the beta 4 integrin at low levels, AMC-HN-3, -4, -7, and -9, have intermediate beta 4 expression, and AMC-HN-8 has extremely high beta 4 expression. The AMC-HN cell lines are representative in vitro models for the study of head and neck cancer biology. Our preliminary results indicate a close relationship between integrin expression and cell adhesion in vitro.     

Obstetric perspectives: consensus of the gynecology department of the University of Liège. Document of the 3rd cycle studies, October 96

Eleven cases of vesicouterine fistula were reported. The initial diagnosis was made when urinary incontinence was occurring. The diagnosis was best confirmed by cystoscopy and hysterography, which was positive in 9 cases. The ultrasonographic examinations allow to visualize the fistula in 5 out of 6 cases examined and to establish the diagnosis. If sonographic examination visualizes the fistula, the performing of hysterography is not necessary. The management consists of the cure of the infection of the urogenital tract and abdominal operation based on the excision of the fistula and closing of the bladder and uterus. All operated patients were cured.     

International roughness index models for HMA overlay treatment of flexible and composite pavements

Timely rehabilitation and preservation of pavement systems are imperative to minimising agency's costs and maximising benefits. Reasonable estimates of treatment life and pavement life extension can be made possible by developing reliable treatment performance models. Louisiana Department of Transportation and Development initiated a three-phased study to develop pavement treatment performance models in support of cost-effective selection of pavement treatment type and the time of treatment. As a result of the study, international roughness index (IRI) models for overlay treatment of composite and flexible pavements were developed. Various factors affecting the IRI of overlay treatment were identified. Climatic indices pertaining to Louisiana were developed which exhibited strong statistical significance along with the other variables as used in the IRI models. The developed IRI models provided good agreement between the measured and predicted IRI values with the majority of data within 5% of prediction error. The models could be used as a good pavement management tool for pavement maintenance and rehabilitation actions.     

Single Event crosstalk shielding for CMOS logic

With advances in technology scaling, CMOS circuits are increasingly more sensitive to transient pulses caused by Single Event particles. Hardening techniques for CMOS combinational logic have been developed to address the problems associated with Single Event transients, but in these designs, Single Event crosstalk effects have been ignored. In order to complement the Single Event upset (SEU) hardening process, coupling effects among interconnects need to be considered in the Single Event hardening and analysis of CMOS logic gates due to technology scaling effects that increase both SE vulnerability and crosstalk effects. As technologies advance, the coupling effects increasingly cause SE transients to contaminate electronically unrelated circuit paths which can in turn increase the “Single Event susceptibility” of CMOS circuits. Serious effects may occur if the affected line is a clock line or an input line of voters in triple-modular redundancy (TMR) circuit. Hence, this work first analyzes Single Event crosstalk on recent technologies and then proposes hardening techniques to reduce Single Event crosstalk. Hardening results are demonstrated using HSpice Simulations with interconnect and device parameters derived in 90 nm technology.