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Results for “Enhancer mapping”

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PubMed2001

Perfusion characteristics of oleic acid--injured canine lung on Gd-DTPA--enhanced dynamic magnetic resonance imaging.

RATIONALE AND OBJECTIVES: We conducted an animal study to describe and interpret the perfusion characteristics of oleic acid (OA)-injured lungs on gadopentetate dimeglumine (Gd-DTPA)-enhanced dynamic perfusion magnetic resonance (MR) imaging. METHODS: Fourteen dogs received an intravenous OA infusion in the supine (n = 4), prone (n = 4), and right lateral decubitus (n = 6) positions, and 10 minutes later these animals in the same postures underwent the dynamic MR study. Regional Gd-DTPA kinetics was analyzed by the time-signal intensity (SI) curves and by qualitative functional map images of the mean transit time that was representative of the mean circulation time in the vascular bed and the average cumulative sum of the relative increases in SI representative of Gd-DTPA distribution volume during Gd-DTPA first pass. The results were compared with those in six control animals and in another six animals that underwent the MR study 3 minutes (n = 3) and 60 minutes (n = 3) after OA infusion. The MR findings were correlated with the distribution of lung damage and the infused OA particles as assessed by histology. RESULTS: The dynamic MR study showed postural shifts on the gravity-dependent perfusion map of normal lungs. Contrast enhancement during Gd-DTPA first pass in the lung was lower and more heterogeneous in the OA-injured lung models than in controls but was followed by conversely greater and persistent enhancement during the Gd-DTPA redistribution phase. Regardless of the postures for OA infusion, these abnormalities were predominant in the dependent lungs and became more pronounced with time after OA infusion, where more prominent capillary obstruction with OA droplets and alveolar/interstitial edema were histologically observed. On the functional map images, greater mean transit time and the average cumulative sum of the relative increases in SI values were also predominantly distributed in the dependent lungs. CONCLUSIONS: Low and heterogeneous enhancement was observed during Gd-DTPA first pass but was followed by persistent enhancement during the Gd-DTPA redistribution phase, and predominant abnormalities in the dependent lungs may be characteristic features of the perfusion of OA-injured lungs. The histological correlations indicate that these abnormalities may reflect OA-induced pathophysiologies associated with capillary OA obstruction, increased vascular resistance, and capillary permeability/extravascular spaces and that lung damage may be gravity dependent.

Animals↗

PubMed1988

Mapping gold-labeled IgE receptors on mast cells by scanning electron microscopy: receptor distributions revealed by silver enhancement, backscattered electron imaging, and digital image analysis.

Immunogold labeling and silver enhancement techniques are widely used to determine density and distribution of cell membrane receptors by light and transmission electron microscopy. However, these techniques have not been widely used for receptor detection by scanning electron microscopy. We used antigen- or protein A-conjugated colloidal gold particles, together with silver enhancement, sequential secondary and back-scattered electron imaging (SEI and BEI), and digital image processing, to explore cell surface distribution of IgE-receptor complexes on RBL-2H3 cells, a rat leukemia line that provides a model for the study of mucosal mast cells. Cells were first incubated with a monoclonal antidinitrophenol IgE (anti-DNP-IgE) that binds with high affinity to cell surface IgE receptors. The resulting IgE-receptor complexes were cross-linked either with the multivalent antigen, DNP-BSA-gold, or with a polyclonal anti-IgE antibody. Antibody-treated cells were labeled after fixation with protein A-gold. Fixed, gold-labeled cell monolayers were silver enhanced (or not), dehydrated, critical point-dried, and coated with gold-palladium (for SEI analysis) or carbon (for combined SEI/BEI analysis). They were observed in an Hitachi S800 SEM equipped with a field emission tip and a Robinson backscattered electron detector. An image processor (MegaVision 1024XM) digitized images directly from the S800 microscope at 500-1000 line resolution. Silver enhancement significantly improves detection of gold particles in both SEI and BEI modes of SEM. On gold-palladium-coated samples, 20-nm particles are resolved by SEI after enhancement. BEI resolves 15-nm particles without enhancement and 5- or 10-nm particles are resolved by BEI on silver-enhanced, carbon-coated samples. Neither BEI nor SEI alone can yield high resolution topographical maps of receptor distribution (BEI forms images on the basis of atomic number contrast which reveals gold but not surface features). Image analysis techniques were therefore introduced to digitize, enhance, and process BEI and SEI images of the same field of view. The resulting high-contrast, high-resolution images were superimposed, yielding well-resolved maps of the distribution of antigen-IgE-receptor complexes on the surface of RBL-2H3 mast cells. The maps are stored in digital form, as required for computer-based quantitative morphometric analyses. These techniques of silver enhancement, combined BEI/SEI imaging, and digital image analysis can be applied to analyze density and distribution of any gold-labeled ligand on its target cell.

Cell Line↗

PubMed1999

MR measurement of regional relative cerebral blood volume in epilepsy.

The purpose of this study was to evaluate the utility of magnetic resonance (MR) relative cerebral blood volume (rCBV) maps for studying regional hemodynamic changes in interictal and ictal epilepsy patients. Ten epilepsy patients were examined on a 1.5 T MR system. Nine patients were investigated interictally and one patient ictally. In the nine interictal patients, the dynamic plane was defined coronally through the hippocampus symmetrically. For the ictal patient, an axial dynamic plane was defined and the patient was scanned during seizure. Positron emission tomography (PET) studies were performed in 8 of the 10 patients. Lower rCBV of the left hippocampus was predicted by rCBV maps in seven of the nine interictal patients. The mean ratios of rCBV were 1.96 for left hippocampus/white matter and 2.49 for right hippocampus/white matter. The difference between these two ratios is statistically significant (P = 0.01, t-test). In two of the nine interictal temporal lobe epilepsy patients, lower rCBV areas were observed in the right hippocampus. In the ictal patient, the regional rCBV map demonstrated increased blood volume in the lesions. In eight of eight patients who underwent PET studies, MR rCBV findings were consistent with PET findings. The results show that regional hemodynamic changes in epilepsy can be evaluated with dynamic contrast-enhanced MR imaging. MR rCBV maps are sensitive to characterize seizure foci both ictally and interictally.

Adult↗

PubMed2006

Protection of glioblastoma cells from cisplatin cytotoxicity via protein kinase Ciota-mediated attenuation of p38 MAP kinase signaling.

Glioblastoma multiforme is an aggressive form of brain cancer that responds poorly to chemotherapy and is generally incurable. The basis for the poor response of this cancer to chemotherapy is not well understood. The atypical protein kinases C (PKCiota and PKCzeta) have previously been implicated in leukaemia cell chemoresistance. To assess the role of atypical PKC in glioblastoma cell chemoresistance, RNA interference was used to deplete human glioblastoma cells of PKCiota. Transfection of cells with either of two different RNA duplexes specific for PKCiota caused a partial sensitisation to cell death induced by the chemotherapy agent cisplatin. To screen for possible mechanisms for PKCiota-mediated chemoresistance, microarray analysis of gene expression was performed on RNA from glioblastoma cells that were either untreated or depleted of PKCiota. This identified sets of genes that were regulated either positively or negatively by PKCiota. Within the set of genes that were negatively regulated by PKCiota, the function of the gene coding for GMFbeta, an enhancer of p38 mitogen-activated protein kinase (MAP kinase) signaling, was investigated further, as the p38 MAP kinase pathway has been previously identified as a key mediator of cisplatin cytotoxicity. The expression of both GMFbeta mRNA and protein increased upon PKCiota depletion, and this was accompanied by an increase in cisplatin-activated p38 MAP kinase signaling. Transient overexpression of GMFbeta increased cisplatin-activated p38 MAP kinase signaling and also sensitised cells to cisplatin cytotoxicity. The increase in cisplatin cytotoxicity seen with PKCiota depletion was blocked by the p38 MAP kinase inhibitor SKF86002. These data show that PKCiota can confer partial resistance to cisplatin in glioblastoma cells by suppressing GMFbeta-mediated enhancement of p38 MAP kinase signaling.

Antineoplastic Agents↗

PubMed1991

Using conceptual matrices, knowledge maps, and scripted cooperation to improve personal management strategies.

Research in the area of substance abuse suggests that ineffective personal management strategies appear to be a major factor in abuse behavior. In this study, fifty-seven students in an intermediate level psychology class were provided with three tools previously found to enhance academic learning strategies--knowledge maps, scripted peer cooperation, and conceptual matrices--as a means of understanding and improving personal management strategies. Student ratings of the value of these activities were significantly higher than an average or moderate response. Individual differences appear to be an important consideration in predicting participants' perceptions of the value of this approach. Those who felt they had control of whatever happens to them, and those who perceived problems as impactful and had strong desires to change recurring problem situations, had the most positive reactions to the types of activities used in this study.

Adult↗

PubMed1999

Breast fibroadenoma: mapping of pathophysiologic features with three-time-point, contrast-enhanced MR imaging--pilot study.

The capability of three-time-point, gadolinium-enhanced magnetic resonance imaging to depict vascular permeability and extracellular volume fraction of breast fibroadenoma was evaluated with histopathologic correlation. This method demonstrated an even distribution of high extracellular volume fraction and low to moderate microvascular permeability in these common breast lesions, providing a nonsurgical means of improving the accuracy of diagnosis of fibroadenoma.

Adult↗

PubMed2004

Mapping of the functional microcirculation in vital organs using contrast-enhanced in vivo video microscopy.

A functional microcirculation is vital to the survival of mammalian tissues. In vivo video microscopy is often used in animal models to assess microvascular function, providing real-time observation of blood flow in normal and diseased tissues. To extend the capabilities of in vivo video microscopy, we have developed a contrast-enhanced system with postprocessing video analysis tools that permit quantitative assessment of microvascular geometry and function in vital organs and tissues. FITC-labeled dextran (250 kDa) was injected intravenously into anesthetized mice to provide intravascular fluorescence contrast with darker red blood cell (RBC) motion. Digitized video images of microcirculation in a variety of internal organs (e.g., lung, liver, ovary, and kidney) were processed using computer-based motion correction to remove background respiratory and cardiac movement. Stabilized videos were analyzed to generate a series of functional images revealing microhemodynamic parameters, such as plasma perfusion, RBC perfusion, and RBC supply rate. Fluorescence contrast revealed characteristic microvascular arrangements within different organs, and images generated from video sequences of liver metastases showed a marked reduction in the proportion of tumor vessels that were functional. Analysis of processed video sequences showed large reductions in vessel volume, length, and branch-point density, with a near doubling in vessel segment length. This study demonstrates that postprocessing of fluorescence contrast video sequences of the microcirculation can provide quantitative images useful for studies in a wide range of model systems.

Animals↗

PubMed2000

Mapping the SF2/ASF binding sites in the bovine growth hormone exonic splicing enhancer.

Splicing of the last intron (intron D) of the bovine growth hormone pre-mRNA requires the presence of a downstream exonic splicing enhancer (ESE). This enhancer is contained within a 115-nucleotide FspI-PvuII (FP) fragment located in the middle of the last exon (exon 5). Previous work showed that the splicing factor SF2/ASF binds to this FP region and stimulates splicing of intron D in vitro. However, the precise sequences recognized by SF2/ASF within the FP region had not been determined. Here we used multiple strategies to map the SF2/ASF binding sites and determine their importance for ESE function. Taking advantage of the fact that SF2/ASF ultraviolet (UV) cross-links specifically to RNA containing the FP sequence, we first mapped a major SF2/ASF binding site by UV cross-linking and reverse transcription. This strategy identified a 29-nucleotide SF2/ASF binding region in the middle of the FP sequence containing the 7-nucleotide purine-rich motif described previously. Interestingly, this binding region is neither sufficient, nor absolutely required for SF2/ASF-mediated splicing, suggesting that additional SF2/ASF binding sites are present. The location of these additional sites was determined by electrophoretic mobility shift analysis of various subfragments of the FP sequence. Antisense 2'-O-methyl oligoribonucleotides complementary to selected SF2/ASF binding sites block bovine growth hormone intron D splicing. Thus, multiple SF2/ASF binding sites within the exonic splicing enhancer contribute to maximal enhancer activity.

Animals↗

PubMed2005

Distribution of Ixodes ricinus in the British Isles: investigation of historical records.

Ixodes ricinus Linnaeus (Acari: Ixodidae) is the most abundant and widely distributed tick in the British Isles, and is a vector for a number of bacterial, viral and protozoal pathogens of both medical and veterinary importance. This report provides an update to the historical distribution data of I. ricinus, published by the Biological Records Centre (BRC), Monks Wood in The Provisional Atlas of the Ticks (Ixodidae) of the British Isles by K. P. Martyn (1988), and is supplemented with additional BRC records since 1988, additional data from published scientific literature and unpublished field studies, and enhanced with spatial and temporal information on tick stages collected and their host associations. Records have been mapped at 10 km resolution and enhanced to 5 km, 1 km and 0.1 km. Differentiation between records representing one-off collections from those representing populations of I. ricinus has been achieved through the classification of the records into either reported or established populations. Detailed seasonality and host associations of records are investigated, highlighting the value in obtaining additional detailed contemporary data to aid risk assessments and research within this field.

Animals↗

PubMed2002

Receptor-selective effects of endogenous RGS3 and RGS5 to regulate mitogen-activated protein kinase activation in rat vascular smooth muscle cells.

Regulators of G protein signaling (RGS) proteins compose a highly diverse protein family best known for inhibition of G protein signaling by enhancing GTP hydrolysis by Galpha subunits. Little is known about the function of endogenous RGS proteins. In this study, we used synthetic ribozymes targeted to RGS2, RGS3, RGS5, and RGS7 to assess their function. After demonstrating the specificity of in vitro cleavage by the RGS ribozymes, rat aorta smooth muscle cells were used for transient transfection with the RGS-specific ribozymes. RGS3 and RGS5 ribozymes differentially enhanced carbachol- and angiotensin II-induced MAP kinase activity, respectively, whereas RGS2 and RGS7 ribozymes had no effect. This enhancement was pertussis toxin-insensitive. Thus RGS3 is a negative modulator of muscarinic m3 receptor signaling, and RGS5 is a negative modulator of angiotensin AT1a receptor signaling through G(q/11). Also, RGS5 ribozyme enhanced angiotensin-stimulated inositol phosphate release. These results indicate the feasibility of using the ribozyme technology to determine the functional role of endogenous RGS proteins in signaling pathways and to define novel receptor-selective roles of endogenous RGS3 and RGS5 in modulating MAP kinase responses to either carbachol or angiotensin.

Animals↗

PubMed2002

Magnetic resonance imaging characterization of hemorrhagic transformation of embolic stroke in the rat.

Intracranial hemorrhage is a critical factor when considering efficacy and safety of thrombolytic intervention after thromboembolic stroke. This study tested whether magnetic resonance imaging could identify tissue for hemorrhagic transformation after the onset of embolic stroke. Rats subjected to embolic stroke with and without recombinant tissue-type plasmogen activator (rt-PA) treatment were followed-up with magnetic resonance imaging using the inverse of the apparent forward transfer rate for magnetization transfer (k(inv)), gadolinium-chelate contrast-enhanced magnetic resonance imaging, and diffusion-, perfusion-, and T2-weighted imaging. Rats with embolic stroke either were treated with rt-PA 1 (n = 16) or 4 hours (n = 13) after stroke onset or were not treated (n = 15). From these groups, at total of 17 rats had intracerebral hemorrhage. Tissue progressing to hemorrhage and adjacent to the site of hemorrhage was analyzed to identify magnetic resonance imaging markers that characterize hemorrhagic transformation. The parameter maps of k(inv) and contrast-enhanced magnetic resonance imaging showed greater sensitivity in the detection of tissue destined for hemorrhagic transformation compared with the apparent diffusion coefficient of water (ADCw) and CBF. In tissue not destined to undergo hemorrhagic transformation, k(inv) maps and contrast-enhanced magnetic resonance imaging exhibited small increases in k(inv) and contrast-enhanced magnetic resonance imaging signal intensity in the area encompassing the territory supplied by the middle cerebral artery. In contrast, large increases in k(inv) and in signal intensity in the contrast-enhanced magnetic resonance images were detected in the region where gross hemorrhage was confirmed histologically. The values of k(inv), T2, and signal intensity in the contrast-enhanced magnetic resonance images were significantly higher in the region destined for hemorrhagic transformation (k(inv), P < or = 0.033 3-24 hours after embolization; T2, P < or = 0.037 24-48 hours; contrast-enhanced magnetic resonance imaging, P < 0.05 4-7 hours) compared with the nonhemorrhagic transformation ischemic region or in the contralateral homologous regions after onset of ischemia. Of these methods, k(inv) shows the most sensitivity in the detection of hemorrhagic transformation soon after embolization. The authors' data suggest that k(inv) and contrast-enhanced magnetic resonance imaging are potentially important methodologies for detecting tissue destined for hemorrhagic transformation.

Animals↗

PubMed1989

Modification of behavioral responses to methamphetamine evoked by the stimulant's metabolite p-hydroxynorephedrine in rats.

The central effect of p-hydroxynorephedrine (OH-NE), one of the p-hydroxylated metabolites of methamphetamine (MAP) and amphetamine (AMP), was investigated in rats. Locomotion and stereotypy were examined after SC injections of 0.5-5 mg/kg of MAP or 0.02-0.5 mg/kg of apomorphine (APO) in animals treated with either saline or 5-50 mg/kg of OH-NE IP 20 hr before behavioral assessment. The locomotor stimulating effect of both 0.5-2 mg/kg of MAP and 0.2 mg/kg of APO was enhanced by 5 mg/kg of OH-NE. On the other hand, 30 mg/kg of OH-NE severely suppressed the stimulating effect of MAP but had no influence on that induced by 0.2 mg/kg of APO. The stereotypy induced by 5 mg/kg of MAP or 0.5 mg/kg of APO was enhanced and prolonged in the OH-NE-treated rats. Subsequently, examinations were performed to determine whether OH-NE had any effect on the dopaminergic mechanism. Hypomotility induced by 0.02 mg/kg of APO was alleviated by 5 mg/kg of OH-NE, but was aggravated by 30 mg/kg. These results suggest that OH-NE administered prior to SC injections of MAP or APO influences their behavioral effects via the dopaminergic mechanism. The possibility that other neural mechanisms may be involved in this OH-NE-induced behavioral modification is also discussed.

Animals↗

PubMed2001

In vivo detection of 13C-enriched glucose metabolites in mouse brain by T-SEDOR imaging.

Protons J-coupled to 13C were selectively detected in the mouse head by in vivo 1H NMR imaging based on Twin Spin Echo DOuble Resonance (T-SEDOR) excitation. This pulse sequence combines a good chemical specificity with high sensitivity, requires no solvent pre-saturation and is well adapted to the imaging modality. 1H T-SEDOR maps of the mouse head allowed detection of areas of preferential accumulation of 13C-enriched compounds, upon repeated injections of uniformly 13C-labelled glucose, which induced hyperglycemia. The results demonstrated the feasibility, both in time scale and metabolite concentration, of applying T-SEDOR MRI for in vivo mapping brain areas characterized by enhanced rates of glucose uptake and/or accumulation of its metabolites.

Animals↗

PubMed2002

Visualizing metabolic activity on a genome-wide scale.

MOTIVATION: To enhance the exploration of gene expression data in a metabolic context, one requires an application that allows the integration of this data and which represents this data in a (genome-wide) metabolic map. The layout of this metabolic map must be highly flexible to enable discoveries of biological phenomena. Moreover, it must allow the simultaneous representation of additional information about genes and enzymes. Since the layout and properties of existing maps did not fulfill our requirements, we developed a new way of representing gene expression data in metabolic charts. RESULTS: ViMAc generates user-specified (genome-wide) metabolic maps to explore gene expression data. To enhance the interpretation of these maps information such as sub-cellular localization is included. ViMAc can be used to analyse human or yeast expression data obtained with DNA microarrays or SAGE. We introduce our metabolic map method and demonstrate how it can be applied to explore DNA microarray data for yeast. AVAILABILITY: ViMAc is freely available for academic institutions on request from the authors.

Computational Biology↗

PubMed1984

Adaptive grey level assignment in CT scan display.

The normal method of assigning grey levels to computed tomographic (CT) numbers in CT scan display involves interactive selection of an intensity window. This method often results in the inability to visualize different types of tissue in a single image and sometimes results in unnecessary loss of contrast sensitivity. A method is described that solves these problems by automatically adapting the assignment of displayable grey levels to CT numbers in a way that varies smoothly across the image according to local needs for the presentation of contrast. The adaptation is based on local histogram equalization implemented using interpolation so as to operate on a minicomputer in a few tens of seconds. This contrast enhancement is followed by a mapping that causes the viewer's perceptual response to be linear across the display scale.

Data Display↗

PubMed1996

Fine mapping of the human and mouse genes for the type I procollagen COOH-terminal proteinase enhancer protein.

The human gene (PCOLCE) for the C-proteinase enhancer, a glycoprotein that enhances the enzymatic activity that cleaves the type I procollagen C-propeptide, was previously mapped to the same cytogenetic region (7q21.3-q22) as COL1A2, the type I collagen pro-alpha 2 chain gene. The apparent proximity of these two genes, whose products interact, might have reflected some mechanism for coregulation of expression. We have employed somatic cell hybrids, cosmid contigs, and interspecific backcross mice to investigate further the apparent proximity of the two genes. Our data show that PCOLCE and COL1A2 are separated by at least 6 Mb on human chromosome 7q and that Pcolce and Col1a2 are located on two separate chromosomes in mouse. These data also indicate that PCOLCE is located within a 10- to 15-cM region of conserved synteny between human chromosome 7 and mouse chromosome 5.

Animals↗

PubMed2001

Homocysteine stimulates the expression of monocyte chemoattractant protein-1 in endothelial cells leading to enhanced monocyte chemotaxis.

Hyperhomocysteinemia has been identified as an independent risk factor for atherosclerosis. The infiltration of monocytes into the arterial wall is one of the key events during atherogenesis. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemokine that stimulates the migration of monocytes into the intima of the arterial wall. The mechanism by which increased monocyte infiltration occurs in atherosclerotic lesions in patients with hyperhomocysteinemia has not been delineated. The objective of the present study was to investigate the effect of homocysteine on MCP-1 production in endothelial cells. Cells were incubated with homocysteine. The secretion of MCP-1 protein was significantly increased (195% as compared to the control) in cells treated with pathological concentrations of homocysteine. Such effect was accompanied by an increased expression of MCP-1 mRNA (176% as compared to the control) in endothelial cells which resulted in enhanced monocyte chemotaxis. The p38 MAP kinase as well as other members of the p38 MAP kinase pathway, including MKK3, MKK6, ATF-2 and Elk-1, were activated in homocysteine-treated cells. Homocysteine-induced MCP-1 expression and subsequent monocyte chemotaxis were blocked by a p38 MAP kinase inhibitor (SB203580) suggesting that the p38 MAP kinase pathway might be involved in homocysteine-induced MCP-1 expression in endothelial cells. In contrast, staurosporine, a protein kinase C inhibitor, had no effect on homocysteine-induced MCP-1 expression. In conclusion, our results indicate that homocysteine stimulates MCP-1 expression in endothelial cells leading to enhanced monocyte chemotaxis.

Activating Transcription Factor 2↗

PubMed1989

[Genome mapping in silver fox. Syntenic genes in Carnivora].

Hamster X fox somatic cell hybrids segregating individual fox chromosomes in different combinations were used to assign seven structural loci to fox chromosomes. The gene for ME1 was mapped on the VFU1 chromosome, the genes for ADK and PP being located on the VFU4 chromosome. The gene for GSR was assigned to the VFU7 chromosome and the genes for MPI and COT1 were assigned to the VFU15 chromosome. Localization of these genes enhances the established fox genetic map and extends the known syntenic homologies between the fox and other mammalian. The comparison of data on gene mapping has provided basis for suggestion that there are significant differences in rates of karyotypic evolution in many mammalian taxa.

Animals↗