Computer-enhanced Image Analysis Research Articles

Regional Distribution Of SDH In Single Muscle Fibres After Training In Endurance Cyclists

Previous research has shown that two populations of mitochondria with different metabolic properties exist in human skeletal muscle. Recent research has shown that the regional distribution of succinate dehydrogenase (SDH) in single muscle fibres is altered with moderate intensity endurance training. Based on this information, we tested the hypothesis that the regional distribution of mitochondria would be altered following sustained high intensity endurance training. PURPOSE The purpose of this study was to examine the changes in the subsarcolemmal (SSM) and intermyofibrillar (IMF) mitochondria as represented by changes in SDH activity. METHODS Eight (2 female, 6 male) trained cyclists (VO2max = 60.7 mL.kg−1·min−1) performed eight weeks of high intensity endurance training for 60 minutes × 5d.wk−1 at 80–85% VO2max. Pre- and post-training muscle biopsy samples were taken from the vastus lateralis, and then immediately frozen in isopentane until analyzed histochemically for SDH activity. Frozen tissue sections (6μm) were incubated and then analyzed using a computer-enhanced image analysis system which digitized selected fibres (18–75) to represent SDH activity as an optical density. Concentric rings (pixel width 0.75μm) were partitioned from the outside layer (SSM, <5.25μm) to the inside layer (IMF, > 6.00μm). Based on myofibrillar ATPase staining, Type I and Type II fibres were identified. RESULTS The results showed that significant changes (ANOVA, p < 0.05) occurred for all analyses performed, both pre- vs. post-training for fibre types, and between SSM and IMF mitochondria. SDH activity increased significantly in Type II (71.7%) vs. Type I (28.3%) over training. The greatest increases in the mitochondrial activity occurred in the SSM (54.6%) vs. IMF (45.4%). However, the greatest increase in SDH activity occurrred in the IMF mitochondria of Type II fibres (39.7%). CONCLUSIONS These data demonstrated that high intensity endurance training can elicit selective adaptations in the metabolic profile of different fibre types in single muscle fibres, and that regional differences in SDH activity can occur.

Adjuvant Effect of Antibodies against von Willebrand Factor, Fibrinogen, and Fibronectin on Staphylokinase-Induced Thrombolysis as Measured Using Mural Thrombi Formed in Rat Mesenteric Venules

The change in thrombus mass during thrombolytic therapy is thought to be the difference between its growth and its degradation induced by thrombolytic agents. Platelets play a pivotal role in arterial thrombosis and bind to each other and to exposed subendothelial matrices via adhesive proteins such as von Willebrand Factor, fibrinogen, and fibronectin. The aim of the present study was to assess whether administration of antibodies against these adhesive proteins, in conjunction with plasminogen activator, could enhance the degradation of platelet-rich thrombus. Mural platelet-rich thrombi were formed in rat mesenteric venules using He–Ne laser irradiation. Recombinant staphylokinase was infused continuously and polyclonal antibodies against adhesive proteins were given by bolus injection. The thrombolytic process was analysed using computer-enhanced image analysis software. Administration of each of the antibodies enhanced staphylokinase-induced thrombolysis.

Determination of the Size of a Foreign Body in the Eye Using Image Analysis of Its Roentgenogram

The case of a computer-enhanced image analysis of X-ray pictures of a pellet in the orbit of a man who sustained a shotgun injury to his right eyeball is presented. The man was shot accidentally during a pheasant hunt. Two hunters were shooting simultaneously but they were using three different sizes of pellet--3.0, 3.5 and 4.0 mm. One of the pellets hit a third man in the eye. He sustained serious injury with a resultant loss of sight in this eye despite immediate medical treatment. Unfortunately the pellet could not be removed from the orbit, but it was necessary to establish who was responsible for the injury. The problem lay in determining the specification and size of the pellet at that moment. The problem was solved using computer-enhanced video image analysis of the X-ray pictures of the pellets. The image processor LUCIA G (http(/)/www.lim.cz) was used.

Lead and mercury mutagenesis: Role of H2O2, superoxide dismutase, and xanthine oxidase

It has been suggested that reactive oxygen intermediates (ROIs) may have a role in the genotoxic effects of lead (Pb2+) and mercury (Hg2+), but there have not been any definitive studies demonstrating a causal relationship between the induction of ROIs by these metals and mutagenesis. We previously demonstrated, using the transgenic Chinese hamster ovary cell line AS52, that low concentrations (0.1-1 microM) of Pb2+ and Hg2+ are mutagenic. In the present study, using a novel histochemical computer-enhanced image analysis technique, we demonstrate that Pb2+ and Hg2+ induce the formation of H2O2 in AS52 cells by at least two distinct mechanisms. One is characterized by the rapid induction of H2O2 following treatment of cells with concentrations of Pb2+ or Hg2+ below 0.8 and 1 microM, respectively, while the second occurs in AS52 cells treated with concentrations of Pb2+ or Hg2+ greater than 0.8 and 1 microM, respectively. Pb2+ and Hg2+ (0.1-1 microM) had no effect on the activities of partially purified catalase, glutathione peroxidase, or glutathione reductase, important enzymes involved with antioxidant defense, but these metals stimulated the activities of copper-zinc superoxide dismutase (CuZn-SOD) and xanthine oxidase (XO). Allopurinol (50 microM), a specific inhibitor of xanthine oxidase, inhibited the induction of H2O2 by Pb2+ (0.8-1 microM) and Hg2+ (1 microM) and also inhibited Pb2+- and Hg2+-induced mutagenesis. These results demonstrate that Pb2+ and Hg2+ disrupt the redox status of AS52 cells by enhancing the activities of CuZn-SOD and XO. Furthermore, the results of these studies also demonstrate that there is a causal relationship between the induction of H2O2 by these metals and mutagenesis.

Role of the mucous surface coat, dietary bulk and mucosal cell turnover in the intestinal disposition of benzo(a)pyrene (BaP)

The dispositon of 3H-benzo(a)pyrene (BaP) equivalents from the diet was examined at the lumen-mucosal cell interface of the proximal, medial and distal regions of the catfish intestine. 3H-BaP (500 μd, 20 μg/kg) was administered via gavage in maintenance diet to two groups of catfish. One group was fed daily after dosing while the other was fasted until collection of intestinal tissues. Autoradiography of cryosectioned tissues and computer-enhanced image analysis allowed examination of BaP disposition. BaP was found to selectively localize at high concentrations in the mucous surface coat and corresponding villi of the colon. Supplemental feeding of uncontaminated diet lowered BaP concentrations in these regions. In a similar but separate experiment, 3H-thymidine (666 μCi) was administered to two groups of catfish by i.p. injection to examine mucosal cell turnover in relation to BaP turnover. 3H-Thymidine incorporation was greatest in the distal regions of both fasted and fed groups. Thymidine dynamics indicated that feeding was related to only marginal changes in cell turnover, which suggests that losses in intravillus BaP with feeding may not be wholly explained by intestinal cell turnover. Results from this study suggest that the mucous surface coat of the intestine is a major factor in the regional disposition of dietary carcinogens and removal of BaP from this mucin layer is related to the intake of dietary bulk.

Evaluation of Six Histological Fixatives Using Image Analysis to Measure Reaction Product Concentration

AbstractThe purpose of this study was two-fold: to determine the effectiveness of manual image analysis (IA) and computer enhanced IA as a quantitative tool to measure reaction products from immunohistological reactions and to determine the effects of different fixatives on the conservation of epitopes when immunostaining for prostate specific antigen.Identical slides were evaluated with both computer enhancer and manual IA. Computer enhanced image analysis showed an overall sensitivity of 84%, was a turn key system and provided more control for measuring parameters than did manual IA. Manual image analysis was inexpensive, sensitive (77%), and practical to adapt to current lab methodologies. Both IA techniques were more sensitive to small but statistically significant differences than either visual or photographic techniques. Image analysis should be incorporated into laboratory protocols to support existing subjective measurements currently in use.The effect on the conservation of epitopes was evaluated...

Evaluation of Six Histological Fixatives Using Image Analysis to Measure Reaction Product Concentration

The purpose of this study was two-fold: to determine the effectiveness of manual image analysis (IA) and computer enhanced IA as a quantitative tool to measure reaction products from immunohistological reactions and to determine the effects of different fixatives on the conservation of epitopes when immunostaining for prostate specific antigen.Identical slides were evaluated with both computer enhancer and manual IA. Computer enhanced image analysis showed an overall sensitivity of 84%, was a turn key system and provided more control for measuring parameters than did manual IA. Manual image analysis was inexpensive, sensitive (77%), and practical to adapt to current lab methodologies. Both IA techniques were more sensitive to small but statistically significant differences than either visual or photographic techniques. Image analysis should be incorporated into laboratory protocols to support existing subjective measurements currently in use.The effect on the conservation of epitopes was evaluated with 6 common histological fixatives (B5, Bouin, Carnoy, Stat Fix, Histochoice, and 10% neutral buffered formalin [NBF]) used to fix prostate tissue collected from surgery. Fixation times for tissues were limited to 4 hr except 10% NBF, which was fixed for hr and from 24–295 hr. Bounin and B5 conserved prostate specific antigen epitope most efficiently as demonstrated photographically and by image analysis. A selected patient trial was repeated to determine if counterstaining or antigen retrieval would affect the ranking of the fixatives. Antigen retrieval was effective in recovering antigenic sites from HistoChoice and Carnoy fixed tissues, but neither antigen retrieval or counterstaining altered the final ranking of fixatives. (The J Histotechnol 18:119, 1995)

Quantitative analysis of lipofuscin and neurofibrillary tangles in the hippocampal neurons of Alzheimer disease brains.

Using computer-enhanced image analysis, the amount of lipofuscin was measured in 500 hippocampal pyramidal neurons (regions CA2 and CA3) with and without neurofibrillary tangles (NFT), in brains of 10 patients with Alzheimer's disease (AD), as well as in 6 age-matched controls. The average content of lipofuscin in those cells from AD brains carrying NFT is only 10% of the total perikaryal area, whereas in the AD neurons free of NFT, and in age matched controls, lipofuscin amounted to 31 and 33% of cellular area, respectively. Measurements of lipofuscin's intrinsic autofluorescence confirmed this material to be three times more abundant in AD neurons without NFT and in controls. We propose that a breakdown in the capacity for making lipofuscin may result in the neuronal inability to store toxic waste. Such a defect could be responsible for the generation of NFT and ultimately may contribute to neuronal demise.

The Use of Deformation Rates in the Scale-Up of Polyethylene Blown Film Extrusion

It is often desirable to scale up a process from a lab scale system, where most research occurs, to a large scale industrial system with minimal ex penses of time and money. This work will show that under appropriate process ing conditions, blown films can be produced under similar strains that yield comparable crystalline morphologies and therefore equivalent mechanical properties. A tracer technique was used to determine deformation in the formation of blown films and applied to both an industrial blown film line and a smaller lab scale analog. Using computer-enhanced image analysis, strain and strain rate data were acquired from the tracer technique. Large and small scale data were compared for similarities in shape and magnitude of respective data plots. Mechanical testing of the films showed that uniaxial and biaxial toughnesses and modulus were similar for paired runs based on a scale-up factor of 1/3 based on output per die circumference. Wide angle X-ray analysis showed struc tural or morphological similarities between films produced under parallel con ditions. The data presented shows that a successful scale-up of polyethylene blown film extrusion was completed using appropriately matched processing condi tions.

Quantifying the pattern of beta/A4 amyloid protein distribution in Alzheimer's disease by image analysis.

We have undertaken a study of the distribution of the beta/A4 amyloid deposited in the cerebral cortex in Alzheimer's disease. Previous studies which have examined the differential distribution of amyloid in the cortex in order to determine the laminar pattern of cortical pathology have not proved to be conclusive. We have developed an alternative method for the solution of this problem. It involves the immunostaining of sections followed by computer-enhanced image analysis. A mathematical model is then used to describe both the amount and the pattern of amyloid across the cortex. This method is both accurate and reliable and also removes many of the problems concerning inter and intra-rater variability in measurement. This method will provide the basis for further quantitative studies on the differential distribution of amyloid in Alzheimer's disease and other cases of dementia where cerebral amyloidosis occurs.

Gastrin-releasing peptide gene products in midtrimester human fetal lung with and without maternal smoking history during pregnancy

A preliminary morphological study on human fetal lungs with positive maternal smoking history demonstrated alterations of the neuroepithelial bodies (NEBs). We studied human fetal lung tissue between the gestational ages of 12 weeks and 19 weeks, comprising 12 cases with a smoking history during pregnancy (Group 1) and eight cases without a smoking history during pregnancy (Group 2). We demonstrated, by immunocytochemistry, the presence of gastrin-releasing peptide (GRP) gene products: GRP 14-27 in all 20 cases, and C terminal peptide of pro-GRP (C-flanking peptide) in 17 cases. Quantification of the neuroepithelial cells (NECs) was made by computer-enhanced image analysis using the Context Vision system, expressing 1) the total stained areas of the NECs per unit area of section and 2) the total staining areas of the NECs per unit area of airway epithelium, measured as the area of cytokeratin immunoreactivity in an adjacent section. The results show no statistically significant difference between groups 1 and 2 for either GRP 14-27 or C-flanking peptides. The apparent lack of influence of maternal smoking during pregnancy on the expression of GRP gene products in the NECs could be a reflection of inherently reduced reactivity of the cells during the gestation period studied. However, a larger series is needed before any conclusions can be made. Alternatively, the adverse effects of smoking might be reflected during the canalicular phase of lung development; an increased immunoreactivity appears to be present during that period. The expression of pro-GRP gene products in the pulmonary NECs of older fetuses and neonates with maternal smoking history during pregnancy requires further study.

A laser microbeam as a tool to introduce genes into cells and organelles of higher plants

AbstractA number of methods have become available to introduce genes into animal cells. In plants the rigid cell wall has presented a substantial barrier to similar efforts. Using protoplasts represents one way to overcome the obstacles to gene transfer. In many species these wall‐less cells, however, are sensitive to manipulations and do not regenerate to plants. Furthermore, direct gene transfer into organelles is even further complicated by the fact that DNA has to pass through two membrane systems. —Recently a method has been developed to deliver DNA randomly to cells with a particle gun [1]. It is very desirable, however, to introduce DNA into specifically selected cells or their organelles. —A laser beam focussed to its diffraction limits and coupled into a microscope provides a tool to cut openings of less than 1 μm into cell walls or membranes. Under visual control cells can be selected for gene transfer or for surgical manipulation [2]. Direct transformation with cloned DNA was achieved in mammalian cells [3,4], —Now this tool was used on oilseed rape (Brassica napus). With a laser microbeam DNA was introduced into cells and microspores of Brassica napus [5,6]. Individual cells were placed in the light path of the microscope and a single laser pulse was released at the cell wall and membrane. At the site of the laser focus wall and membrane were opened. At the time of the experiment the cells were incubated in a hypotonic buffer containing DNA. The nucleic acid was taken up into the cells. The opening in the membrane was only temporary and closed within less than 5 s after the laser pulse. The cells or microspores developed into colonies or embryos respectively with a frequency of 30–50% of the untreated controls. —Using the same technique DNA was also integrated into isolated chloroplasts [7]. Furthermore DNA can be incorporated into chloroplasts inside of a cell, large quantities of DNA were first brought into the cytoplasma. Then the laser beam was focussed on each chloroplast inside of the cell and a single laser pulse was released at each organelle. Again a small opening was cut into the membrane allowing the DNA to enter the organelle [6]. This opening closed again within 1.2 s as could be demonstrated by computer enhanced image analysis. —Laser microbeams provide a new powerful tool for introducing DNA into plant cells and their organelles as well as for analyzing chromosomes.

Computer-Enhanced Image Analysis of Bacterial Polypeptide Patterns on Two-Dimensional Polyacrylamide Gels

Moline, H. E., and Hruschka, W. R. 1987. Computer-enhanced image analysis of bacterial polypeptide patterns on two-dimensional polyacrylamide gels. Phytopathology 77:,745-747. Comparisons of two-dimensional polyacrylamide gel electrophoresis species specific. Although all bacterial strains studied shared a number of (2-D PAGE) profiles of Erwinia carotovora pv. carotovora, E. c. pv. polypeptides, significant differences were observed between E. c. pv. atroseptica, E. chrysanthemi, E. rhapontici, E. herbicola, and E. amylovora carotovora, E. c. pv. atroseptica, and other species analyzed, allowing were made by transferring gel images to a Hewlett-Packard 1000F differentiation. Computer-enhanced image analysis increased the minicomputer interfaced to a Grinnell 270 image processor. Computer- sensitivity of protein comparisons on the 2-D PAGE gels more than enhanced image analysis produced gel maps that revealed polypeptides threefold over visual comparisons. common to all bacterial strains studied, as well as polypeptides that were

Nuclear pores and interphase chromatin: high-resolution image analysis and freeze etching.

Computer-enhanced analysis of electron micrographs of thin-sectioned rat liver nuclei, combined with three-dimensional reconstruction of the same Feulgen-stained nuclei, points to a unique clustering of chromatin DNA fibres near the nuclear border. Computer-enhanced image analysis has been applied to electron micrographs of the envelopes of the same rat liver nuclei prepared by freeze etching and a few essential geometrical parameters characterizing the pores and their distribution have been determined. During interphase, clusters of nuclear pores, closely paralleling the clustering of membrane-attached chromatin fibres, have been identified on the envelope, the number of these being similar to the number of homologus pairs of metaphase chromosomes. Furthermore, rapid changes induced in chromatin distribution appear to be associated with rapid changes in pore number, but not in the number of pore clusters.