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Demonstration of the relationship between heart dimensions and QRS voltage amplitude.

This study was undertaken to elucidate the still debated question of the relationship between cardiac volume and QRS voltage amplitude. The authors studied 14 healthy men, aged 24-61 years (mean age, 41.2 +/- 12.1 years). They underwent a reduction in venous return, produced by simultaneously inflating sphygmomanometric cuffs placed around the most proximal portion of each of the four limbs. In basal conditions and 5 minutes after cuff inflation, two-dimensional and M-mode echocardiograms were recorded with vectorcardiographic loops and scalar Frank leads. The reduction of the venous return to the heart induced a significant decrease of the end-diastolic left ventricular diameter (from 52.4 +/- 4.2 to 48.5 +/- 4.6 mm, P < .001), of the R wave amplitude in leads X and Y, of the sum of the R wave amplitudes in the three leads,and of the maximal vector in the frontal and horizontal planes. No significant changes in the heart rate or arterial blood pressure were observed. These results support Brody's theory concerning the relationship between cardiac blood volume and QRS voltage.

Adult↗

Rabbit intestinal and colonic mucins: isolation, partial characterization, and measurement of secretion using an enzyme-linked immunoassay.

Mucin was purified separately from the upper, mid, and distal small intestine and the proximal colon of the rabbit. The carbohydrate profiles of the three intestinal mucins were the same but differed from that of the colonic mucin, which contained less N-acetylgalactosamine but more sialic acid. All four mucins had similar polymeric structures composed of large, heterogeneous glycoprotein monomers and a smaller protein of Mr approximately 120,000, held together by disulphide bonds. The three intestinal mucins, however, were more resistant to dissociation by thiol reduction or degradation by proteolysis than colonic mucin. An antibody against a pool of the purified mucins was developed in guinea pigs and used to establish an enzyme-linked immunosorbent assay (ELISA). The antibody was highly specific for rabbit intestinal and colonic mucins, showing no cross-reactivity with nonmucin components of rabbit intestinal and colonic tissue, and very little or no reactivity with purified intestinal mucins from other species. The four purified rabbit mucins had the same affinity for the antibody and similar numbers of antigenic determinants. Antigenicity was apparently associated with the protein moeity of the mucins and was critically dependent on three-dimensional conformation, since proteolysis decreased the number of antigenic determinants while thiol reduction abolished antigen-antibody affinity. Using the ELISA, the tissue mucin content and the rate of mucin secretion were found to be significantly higher in the proximal colon that in the three regions of the intestine, which were the same.

Animals↗

Left ventricular geometry and function in experimental heart failure.

The shape of the left ventricle represents a key element in the adaptation of the heart to a variety of stresses. Therefore, hemodynamic deterioration should be reflected in alterations of shape. To investigate this hypothesis numerical descriptors of shape (single plane, two dimensional shape index, [4 pi (area)/(perimeter)2] and multiplane, three dimensional shape index, [36 pi (volume)2/(surface area)3] were obtained from two dimensional echocardiographic studies in dogs at baseline and then serially during development of early cardiomyopathy. Serial weekly injection of adriamycin (1 mg/kg) in 11 dogs to a cumulative dose of 144 mg/m2 resulted in no significant reduction in ejection fraction, stroke volume or cardiac index (P less than 0.05) but there was demonstrable loss of the ability of the heart to maintain an elliptical configuration during systole. Continuation of treatment to 244 mg/m2 resulted in gradual reduction in ejection fraction, stroke volume and cardiac index. Statistical correlation of the degree of reduction in ejection fraction with change in shape from diastole to systole (change in two and three dimensional shape indices) were -0.803 and -0.855, respectively. These data support the concept that the shape of the left ventricle relates to function, and the inability of the heart to develop an elliptical shape during systole may be an early indicator of impaired function prior to reduction in ejection fraction, stroke volume or cardiac index.

Animals↗

Expansion of chondrocytes in a three-dimensional matrix for tracheal tissue engineering.

BACKGROUND: The generation of autologous tracheal implants by tissue-engineering techniques is a promising concept for otherwise untreatable patients. A functional cartilaginous backbone represents a prerequisite for any bioartificial tracheal graft. The aim of this study was to define suitable cell types and culture conditions for the generation of tracheal cartilage. METHODS: We obtained tracheal, costal, and auricular cartilage from porcine donor animals (n = 10). The chondrocytes were cultured two-dimensionally in cell flasks or mixed with a liquid collagen solution forming a three-dimensional culture system. Labeling with carboxy fluorescein diacetate succinimidyl ester (CFDA SE) and biochemical reduction of formazan served to determine cell viability and proliferation. The extracellular matrix produced by the chondrocytes was characterized by Western blot. RESULTS: The CFDA SE labeling proved viability and the MTT assays documented a proliferation of the chondrocytes over time in vitro. While the chondrocytes in the three-dimensional cell culture system produced hyaline cartilage composed of collagen II, the two-dimensional culture conditions resulted in nonspecific collagen synthesis. CONCLUSIONS: Chondrocytes grown in a three-dimensional matrix can effectively proliferate and produce cartilage and are viable for more than 2 weeks. Costal chondrocytes are suitable for tracheal cartilage tissue engineering.

Animals↗

Regulation of mitotic function of Chk1 through phosphorylation at novel sites by cyclin-dependent kinase 1 (Cdk1).

Chk1 is phosphorylated at Ser317 and Ser345 by ATR in response to stalled replication and genotoxic stresses. This Chk1 activation is thought to play critical roles in the prevention of premature mitosis. However, the behavior of Chk1 in mitosis remains largely unknown. Here we reported that Chk1 was phosphorylated in mitosis. The reduction of this phosphorylation was observed at the metaphase-anaphase transition. Two-dimensional phosphopeptide mapping revealed that Chk1 phosphorylation sites in vivo were completely overlapped with the in vitro sites by cyclin-dependent protein kinase (Cdk) 1 or by p38 MAP kinase. Ser286 and Ser301 were identified as novel phosphorylation sites on Chk1. Treatment with Cdk inhibitor butyrolactone I induced the reduction of Chk1-S301 phosphorylation, although treatment with p38-specific inhibitor SB203580 or siRNA did not. In addition, ionizing radiation (IR) or ultraviolet (UV) light did not induce Chk1 phosphorylation at Ser317 and Ser345 in nocodazole-arrested mitotic cells. These observations imply the regulation of mitotic Chk1 function through Chk1 phosphorylation at novel sites by Cdk1.

Amino Acid Sequence↗

Surface labeling of human platelets by reductive methylation.

A simple, reproducible method for the specific labeling of the surface proteins of human platelets with tritium is described. The labeled platelets were analyzed by two-dimensional gel electrophoresis (isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis) followed by fluorography and the results compared with those obtained by conventional methods. Reductive methylation gave an intense labeling of membrane glycoproteins. There was no cross-linking of the labeled membrane proteins by formaldehyde nor could clear evidence be found that inner proteins were labeled by permeation of the reagents through the plasma membrane. As well as previously described platelet membrane glycoproteins, several others could be detected that were not invariable seen using labeling techniques.

Actins↗

Modeling hexavalent chromium removal in a Bacillus sp. fixed-film bioreactor.

A one-dimensional diffusion-reaction model was developed to simulate Cr(VI) reduction in a Bacillus sp. pure culture biofilm reactor with glucose as a sole supplied carbon and energy source. Substrate utilization and Cr(VI) reduction in the biofilm was best represented by a system of (second-order) partial differential equations (PDEs). The PDE system was solved by the (fourth-order) Runge-Kutta method adjusted for mass transport resistance using the (second-order) Crank-Nicholson and Backward Euler finite difference methods. A heuristic procedure (genetic search algorithm) was used to find global optimum values of Cr(VI) reduction and substrate utilization rate kinetic parameters. The fixed-film bioreactor system yielded higher values of the maximum specific Cr(VI) reduction rate coefficient and Cr(VI) reduction capacity (kmc = 0.062 1/h, and Rc = 0.13 mg/mg, respectively) than previously determined in batch reactors (kmc = 0.022 1/h and Rc = 0.012 mg/mg). The model predicted effluent Cr(VI) concentration well with 98.9% confidence (sigmay2 = 2.37 mg2/L2, N = 119) and effluent glucose with 96.4 % confidence (sigmay(w)2 = 5402 mg2/L2, N = 121, w = 100) over a wide range of Cr(VI) loadings (10-498 mg Cr(VI)/L/d).

Algorithms↗

Electrochemical analysis of the alterations in copper pigments using charge transfer coefficient/peak potential diagrams. Application to microsamples of baroque wall paintings attached to polymer film electrodes.

The alteration of copper pigments in art samples was studied by linear scan and cyclic voltammetry using sample-modified Elvacite 2044 film electrodes on the basis of two-dimensional diagrams of charge transfer coefficients calculated from Tafel plots of reductive dissolution processes vs. peak potential. Characteristic voltammetric peaks were obtained for pigments used in the baroque vault frescoes of the Basílica de la Virgen de los Desamparados painted by Antonio Palomino. Results obtained by voltammetric techniques were compared with those from SEM/EDX and FT-IR analysis obtaining a good agreement and leaving to an unambiguous identification of pigments used by Palomino and their alteration products.

Journal Article↗

Tracking the neurodegeneration of parkinsonian disorders--a pilot study.

The purpose of the study was to explore the possibilities of using diffusion tensor imaging (DTI) and tractography (DTT) for the differential diagnosis and monitoring of disease progression in idiopathic Parkinson's disease (IPD), compared with the atypical parkinsonian disorders multiple system atrophy (MSA) and progressive supranuclear palsy (PSP). A 3.0-T MR scanner was used. DTI was acquired using a single-shot EPI sequence with diffusion encoding in 32 directions and a voxel size of 2 x 2 x 2 mm3. DTI data were analysed and DTT was performed using the PRIDE fibre tracking tool supplied by the manufacturer. The fractional anisotropy (FA) and apparent diffusion coefficient (ADC) within each tract were determined. DTI and DTT images in patients with moderate to advanced MSA demonstrated degeneration of the middle cerebellar peduncles and pontine crossing tracts, with decreased FA and increased ADC. This accounted for most of the pontine and cerebellar atrophy characteristic of this disease. In contrast, patients with PSP showed a selective degeneration of the superior cerebellar peduncle. Three-dimensional images of whole-brain white matter tracts demonstrated a reduction of cortical projection fibres in all patients with PSP. Visualization of the selective degeneration of individual fibre tracts, using DTI and DTT, adds qualitative data facilitating the differential diagnosis of parkinsonian disorders. Repeated measurements of FA and ADC values in a whole fibre tract might be used for monitoring disease progression and studying the effect of treatment in neuroprotective trials. The results are preliminary considering the small number of subjects in the study.

Adult↗

Parameterization of left ventricular wall motion for detection of regional ischemia.

While qualitative wall motion analysis has proven valuable in clinical cardiology practice, quantitative analyses remain too time-consuming for routine clinical use. Our long-term goal is therefore to develop automated methods for quantitative wall motion analysis. In this paper, we utilize a finite element model of the regionally ischemic canine left ventricle to demonstrate a new approach based on parameterization of the left ventricular endocardial surface in prolate spheroidal coordinates. The parameterization provided a substantial data reduction and enabled simple definition, calculation, and display of three-dimensional fractional shortening (3DFS), a quantitative measure of wall motion analogous to the fractional shortening measure used in 2D analysis. The endocardial surface area displaying akinesis or dyskinesis by 3DFS corresponded closely to simulated ischemic region size and 3DFS identified appropriate wall motion abnormalities during experimental coronary occlusion in a canine pilot study. 3DFS therefore appears to be a reasonable candidate for clinical tests to determine its utility in identifying and quantifying acute regional ischemia in patients. By linking state of the art finite element models to the clinically relevant framework of wall motion analysis, the methods presented here will facilitate formulation, in silico prescreening, and clinical testing of additional candidate measures of wall motion.

Animals↗

Response of isolated nuclei to phospholipid vesicles: a two dimensional gel electrophoresis analysis of H2SO4 soluble nuclear proteins after treatment with phosphatidylserine.

H2SO4 soluble proteins extracted from nuclei incubated with phosphatidylserine multilamellar vesicles (PS MLV) have been analysed by means of two-dimensional gel electrophoresis with particular attention to the uH2A fraction. A reduction of H1, H1 degrees and proteins A5, B7, B15 and B23 has been observed in lipid treated nuclei, while the core histones, as well as uH2A are unaffected by liposome treatment. Since these proteins show in vitro the same binding affinity for PS, their behaviour appears to be related to difference in localization in the nucleosome, responsible for their variable accessibility in the chromatin. These results might explain how this phospholipid induces a decondensation of chromatin and a stimulation of RNA synthesis.

Animals↗

Comprehensive two-dimensional gas chromatography: metrics, potentials, limits.

Metrics for evaluation of separation performance of comprehensive two-dimensional gas chromatography (GCxGC) and for quantitative comparison of that performance with similar performance of its 1D (one-dimensional) counterparts are described. The performance improvement can be expressed via reduction in the saturation of a chromatogram or-in the case of the uniform distribution of peaks along the second dimension--via the peak capacity gain due to GCxGC. An order of magnitude peak capacity gain due to the comprehensive GCxGC is possible under optimal conditions. Optimal parameters of the second dimension column as well as the optimal operational conditions for that column and for the modulator in a comprehensive GCxGC are also presented.

Chromatography, Gas↗

Micromachined ultrasound transducers with improved coupling factors from a CMOS compatible process

For medical high frequency acoustic imaging purposes the reduction in size of a single transducer element for one-dimensional and even more for two-dimensional arrays is more and more limited by fabrication and cabling technology. In the fields of industrial distance measurement and simple object recognition low cost phased arrays are lacking. Both problems can be solved with micromachined ultrasound transducers (MUTs). A single transducer is made of a large number of microscopic elements. Because of the array structure of these transducers, groups of elements can be built up and used as a phased array. By integrating parts of the sensor electronics on chip, the cabling effort for arrays can be reduced markedly. In contrast to standard ultrasonic technology, which is based on massive thickness resonators, vibrating membranes are the radiating elements of the MUTs. New micromachining technologies have emerged, allowing a highly reproducible fabrication of electrostatically driven membranes with gap heights below 500 nm. A microelectronic BiCMOS process was extended for surface micromechanics (T. Scheiter et al., Proceedings 11th European Conference on Solid-State Transducers, Warsaw, Vol. 3, 1997, pp. 1595-1598). Additional process steps were included for the realization of the membranes which form sealed cavities with the underlying substrate. Membrane and substrate are the opposite electrodes of a capacitive transducer. The transducers can be integrated monolithically on one chip together with the driving, preamplifying and multiplexing circuitry, thus reducing parasitic capacities and noise level significantly. Owing to their low mass the transducers are very well matched to fluid loads, resulting in a very high bandwidth of 50-100% (C. Eccardt et al., Proceedings Ultrasonics Symposium, San Antonio, Vol. 2, 1996, pp. 959-962; P.C. Eccardt et al., Proceedings of the 1997 Ultrasonics Symposium, Toronto, Vol. 2, 1997, pp. 1609-1618). In the following it is shown how the BiCMOS process has been modified to meet the demands for ultrasound generation and reception. Bias and driving voltages have been reduced down to the 10 V range. The electromechanical coupling is now almost comparable with that for piezoelectric transducers. The measurements exhibit sound pressures and bandwidths that are at least comparable with those of conventional piezoelectric transducer arrays.

Journal Article↗

Computed tomographic analysis of airway dimensions after carotid endarterectomy.

Airway obstruction is a rare but serious postoperative complication of carotid endarterectomy. We prospectively studied airway dimensions between the hyoid bone and cricoid cartilage in patients undergoing carotid endarterectomy using preoperative and postoperative computed tomographic (CT) scans of the neck. CT scans showed soft tissue swelling in all 19 patients. Five patients with clinical evidence of airway obstruction and a hematoma present on CT scan were intubated postoperatively. The three-dimensional reconstruction of the neck from the CT scans showed a reduction in the volume of the airway in all patients. This reduction was greater in the intubated (62% +/- 9%) compared to the nonintubated (32% +/- 7%) patients (P < 0.01). The anterior-posterior and transverse diameters of the airway were reduced, while retropharyngeal edema was increased after carotid endarterectomy. This change was greater for the upper airway at the level of the hyoid compared to the arytenoids and cricoid, and was significantly greater in the intubated than the nonintubated patients. Tracheal deviation was greater in the intubated than in the nonintubated patients. These results demonstrate significant soft tissue edema of the neck after carotid endarterectomy that reduces airway volume and can result in airway obstruction.

Aged↗

Detection of mild coronary stenoses using the Dynamic Spatial Reconstructor.

The accuracy of the Dynamic Spatial Reconstructor (DSR) in the detection of moderate coronary artery stenoses was examined in 20 closed-chest dogs. Twenty-eight hollow plastic cylinders were embolized into the left coronary arteries and produced 25% to 56% reductions in arterial lumina diameter. For each dog, one three-dimensional (3-D) image of the heart was reconstructed from each DSR scan recorded during injection of contrast into the aortic root. Analysis involved blinded visual analysis by four independent observers of multiview projection images computed from the single 3-D image. Postmortem coronary angiograms of the isolated heart were considered definitive for location of the stenoses. Overall sensitivity of detection by DSR was 89% and specificity 81%. Sensitivity of detecting stenoses greater than or equal to 50% was 98%. Receiver operating characteristics (ROC) analysis showed that detection of stenoses in the left coronary arteries is of equal sensitivity and specificity.

Animals↗

Phase space deformation and basis set optimization.

By deforming a given region of phase space---occupied by some unknown eigenfunctions one wishes to find---into a standard, integrable region, effective reductions in basis set size can be achieved. In one-dimensional problems we are able to achieve B/C=1+O(Planck' s constant), where B is the basis set size and C is the number of "converged" eigenfunctions. This result is confirmed by numerical examples, which also indicate exponential convergence as the basis set size is increased. In higher dimensions we prove that such an optimistic result is impossible; we expect that the best one can do in this case is B/C=a+o(1), where a>1 has a geometric interpretation in terms of ratios of volumes in phase space.

Journal Article↗

Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast.

The developmental relationships between the different mammary epithelial cell lineages in the human mammary gland are not well defined. To characterize human breast epithelial cells (HBEC) with progenitor activity, we used flow cytometry and single cell sorting to analyze the distribution of cellular phenotypes in primary cultures of reduction mammoplasties and their associated ability to generate colonies in 2-dimensional (D) and 3-D (collagen gel) culture systems. This approach allowed two distinct types of HBEC progenitor populations to be distinguished on the basis of their differential expression of the MUC-1 glycoprotein, CALLA/CD10 and epithelial-specific antigen (ESA). The first type of progenitor, which is enriched in the MUC-1+/CAL-LA-/ESA+ subpopulation, generated colonies of tightly arranged cells in 2-D cultures and small alveolar-like colonies with a central lumen when cultured in a collagen matrix. The cells produced in the colonies and derived from these MUC-1+/CALLA-/ESA+ progenitors were found to express typical luminal epitopes (keratin 8/18, keratin 19, MUC-1, ESA) and showed low levels of expression of myoepithelial epitopes (keratin 14 and CD44v6). The second type of progenitor, which is present in the MUC-1-to +/-/CALLA +/- to +/ESA+ subpopulation, generated mixed colonies of both luminal and myoepithelial cells when seeded in 2-D and 3-D cultures. In 2-D cultures, the centrally located cells exhibited a luminal morphology and expressed ESA, but were heterogeneous in their expression of MUC-1. Radiating from the periphery of these ESA+ HBEC were highly refractile ESA- teardrop-shaped myoepithelial-like cells. When cultured in a collagen matrix, these bipotent progenitors generated large branched colonies composed of a heterogeneous population of cells, with some of the progeny cells expressing luminal epitopes (keratin 8/18, keratin 19 and MUC-1) and others expressing myoepithelial epitopes (keratin 14 and CD44v6). A third type of progenitor, which became apparent is passaged HBEC cultures and was enriched in the MUC-1-/CALLA+/ESA- subpopulation, was found to generate colonies of cells with an exclusively myoepithelial phenotype. These results provide definitive evidence for the existence of multilineage HBEC progenitors in normal adult human mammary tissue. The phenotypic profile of these cells suggest that these multilineage progenitors are a relatively undifferentiated cell since they express low levels of MUC-1 and that they have a luminal location within the mammary epithelium since they are ESA+. Furthermore, we suggest that the MUC-1+/CALLA-/ESA+ and the MUC-1- to +/-/CALLA +/- to +/ESA+ progenitors we have identified and characterized are candidate in vivo alveolar and ductal progenitors, respectively.

Actins↗

Transmission loss predictions for dissipative silencers of arbitrary cross section in the presence of mean flow.

A numerical technique is developed for the analysis of dissipative silencers of arbitrary, but axially uniform, cross section. Mean gas flow is included in a central airway that is separated from a bulk reacting porous material by a concentric perforate screen. The analysis begins by employing the finite element method to extract the eigenvalues and associated eigenvectors for a silencer of infinite length. Point collocation is then used to match the expanded acoustic pressure and velocity fields in the silencer chamber to those in the inlet and outlet pipes. Transmission loss predictions are compared with experimental measurements taken for two automotive dissipative silencers with elliptical cross sections. Good agreement between prediction and experiment is observed both without mean flow and for a mean flow Mach number of 0.15. It is demonstrated also that the technique presented offers a considerable reduction in the computational expenditure when compared to a three-dimensional finite element analysis.

Journal Article↗