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At least 919 records · Page 51Linked to original sources
PubMed2002

Proteomic approaches for the global analysis of proteins.

Improvements in technology that allow miniaturization and high-throughput analyses of thousand of genes and gene products have changed the focus and scope of research and development in both academia and industry. It is now possible to study entire proteomes with the goals of elucidating protein expression, subcellular localization, biochemical activities, and their regulation. Alterations in different cell types and conditions and in normal and disease states can be revealed. This wealth of information not only has facilitated our basic understanding of many biological processes but also has enormous potential for drug discovery and development.

Data Collection↗
PubMed2006

Endogenous inhibitory cytokines repress TNFalpha secretion.

Tumor necrosis factor alpha (TNFalpha), with the potential to destroy tissue, is likely to be tightly regulated. A major regulatory step is the translational repression of TNFalpha. This study evaluates whether endogenous inhibitory cytokines account for this repression. Two cell populations were isolated from peripheral blood using techniques that minimized activation, one composed primarily of monocytes and the other containing T-cells and NK-cells. When cultured without a stimulus in the presence of Abs neutralizing IL-4, IL-10, or TGFbeta, each population released large amounts of TNFalpha, reaching levels induced by PHA or LPS. Their actions were at the post-translational level since the numbers of transcripts did not change, and inhibitors of protein or RNA synthesis had no effects. When inhibitors of 38 MAP kinase and ERK were added, T-cell release of TNFalpha proved to involve both pathways while monocytes were dependent on p38 but not ERK. Changes in soluble TNF receptor levels or cell uptake of TNFalpha were not involved. This study shows that low TNFalpha secretion by resting T-cells and monocytes is maintained by endogenous inhibitors that suppress post-translational processing of TNFalpha by MAP kinases. Keeping TNFalpha levels low is critical to the non-inflammatory steady-state.

Cytokines↗
PubMedDate not supplied

[Radiation mapping as one of main methods of creating high resolution maps of human and animal genomes].

Radiation hybrid mapping (RH mapping) is considered as one of the main methods of constructing physical maps of mammalian genomes. In introduction, theoretical prerequisites of developing of the RH mapping and statistical methods of data analysis are discussed. Comparative characteristics of universal commercial panels of the radiation hybrid somatic cells (RH panels) are shown. In experimental part of the work, RH mapping is used to localise nucleotide sequences adjacent to NotI sites of human chromosome 3 with the aim to integrate contig map of NotI clones to comprehensive maps of human genome. Five nucleotide sequences adjacent to the sites of integration of papilloma virus in human genome and expressed in the cells of cervical cancer were localised. It was demonstrated that the region 13q14.3-q21.1 was enriched with nucleotide sequences involved in the processes of oncogenesis. RH mapping can be considered as one of the most perspective applications of the modern radiation biology in the field of molecular genetics, that is, in constructing physical maps of mammalian genomes with high resolution level.

Animals↗
PubMed2005

Modularity and intersection of "what", "where" and "how" processing of visual stimuli: a new method of FMRI localization.

Research on the modularity of perceptual and cognitive processes has often pointed to a ventral-dorsal distinction in cortical pathways that depend upon the nature of the stimuli and the task. However, it is not clear whether the dorsal, occipital-parietal stream specializes in locating visual objects (i.e., a "where" stream), or taking action toward objects (i.e., a "how" stream), although there is some consensus for a ventral, occipital-temporal "what" stream that specializes in the identification of visual objects. It is also not clear to what extent word and picture processing are modular along these streams, as functional imaging maps to date have not addressed the modularity question directly. Here we present two types of functional imaging maps that directly show modularity and intersection of processing function for word and picture stimuli in tasks that require decisions about "what is", "where is", or "how do you interact with" a stimulus (N=6 participants). Our results reveal a middle dorsal "how" stream with some modular regions of activation that are distinct from activation during "where" processing, and that words and pictures involve several modular regions of activation along these streams.

Adult↗
PubMed1997

Noncontact endocardial mapping: reconstruction of electrograms and isochrones from intracavitary probe potentials.

INTRODUCTION: Mapping endocardial activation and repolarization processes is critical to the study of arrhythmias and selection of therapeutic procedures. Previously, we developed methodology for reconstructing endocardial potentials from potentials measured with a noncontact, intracavitary probe. This study further develops and evaluates the ability of the approach to provide detailed information on the spatiotemporal characteristics of the activation process. Specifically, we reconstructed endocardial electrograms and isochrones throughout the activation process over the entire endocardium during a single beat. METHODS AND RESULTS: Cavity potentials were measured with a 65-electrode probe placed inside an isolated canine left ventricle. Endocardial potentials were measured simultaneously using 52 electrodes. Potentials were acquired during subendocardial pacing from different locations. Computed electrograms at various sites closely resemble the measured electrograms (correlation coefficient > 0.9 at 60% of the electrodes). Computed isochrones locate subendocardial pacing sites with 10-mm accuracy. Two pacing sites, 17 mm apart, were resolved. Critical regions, such as areas of isochrone crowding, were accurately reconstructed. CONCLUSIONS: Results indicate the applicability of the approach to mapping the cardiac excitation process on a beat-by-beat basis without occluding the ventricle. The ability of locating electrical events (e.g., single or multiple initiation sites) is demonstrated. Importantly, the method is shown to be capable of reconstructing electrograms over the entire endocardium and determining nonuniformities of activation spread (e.g., areas of slow conduction). These capabilities are important to clinical application in the electrophysiology laboratory and experimental studies of arrhythmias in the intact animal.

Animals↗
PubMed1976

The expression of normal ventricular repolarization in the body surface distribution of T potentials.

Isopotential maps from 120 normal subjects were obtained from 192 simultaneously recorded electrocardiographic leads. Maps were plotted at 1 msec intervals during the QRS and 5 msec intervals during the ST-T deflection. Repetition of QRS features was evident during all but the first few msec of the initial half of serial T maps. This suggests similarities of the normal sequence of ventricular excitation and recovery. Such similarities have been demonstrated by direct studies but are not evident from other electrocardiographic examinations. Serial maps during later portions of the T wave showed decreasing intensity of potentials with little change of body surface locations. This also correlates with an established feature of ventricular repolarization, namely that potential difference boundaries with stable locations are widely distributed during part of that process. Findings suggest isopotential maps show features of ventricular recovery not apparent from less extensive examinations.

Action Potentials↗
PubMed1992

Instrumentation of a high-shear mixer: evaluation and comparison of a new capacitive sensor, a watt meter, and a strain-gage torque sensor for wet granulation monitoring.

A high-shear mixer was instrumented with a new capacitive sensor, a watt meter, and a strain-gage torque sensor. The output from the capacitive sensor was split into two channels, which monitor both resistive and capacitive changes during the granulation process. The outputs of the devices were related to properties of the wet granulation. The percentage moisture content related linearly to the amplitude channel response. Yield values provided a rheological property to relate with power consumption and torque measurement. Power consumption and torque furnished a similar map of the granulation process. The amplitude channel of the capacitive sensor appears to be more clearly differentiated between binder levels in hydrous lactose-HPMC granulations than either power consumption or torque measurement, based on particle size distributions.

Chemistry, Pharmaceutical↗
PubMed1991

Recent advances in the study of electroreception.

Recent studies on electroreception in fish have focused on the structure and function of recurrent descending pathways, efference copy mechanisms, and multiple neuronal maps involved in the processing of sensory information. Studies on a neuronal oscillator have revealed that different neuronal inputs modulate the pattern of oscillations to produce different forms of behavioral output.

Animals↗
PubMed2001

Stereopsis-processing regions in the human parieto-occipital cortex.

We performed fMRI on the human parieto-occipital cortex in order to identify the neural processing regions of stereopsis. Visual stimulation for stereopsis was performed with a random-dot stereogram displayed in the image guides of a new binocular visual stimulation device that we developed. Interestingly, regions from the dorsal portion of the occipital lobe to the superior parietal lobule were activated by binocular disparity, while the inferior parietal lobule was not activated. Moreover, these regions were shown as dominant in the right hemisphere. Functional brain mapping revealed that the processing regions of stereopsis were dorsally located in parieto-occipital cortex, and that the superior parietal lobule is an important region for neural processing of human stereopsis.

Adult↗
PubMed2004

Evaluation of sLORETA in the presence of noise and multiple sources.

The standardized Low Resolution Brain Electromagnetic Tomography method (sLORETA) can be used to compute statistical maps from EEG and MEG data that indicate the locations of the underlying source processes with low error. These maps are derived by performing a location-wise inverse weighting of the results of a Minimum Norm Least Squares (MNLS) analysis with their estimated variances. In this contribution, we evaluate the performance of the method under the presence of noise and with multiple, simultaneously active sources. It is shown that the sLORETA method localizes well, as compared to other linear approaches such as MNLS and LORETA. However, simultaneously active sources can only be separated if their fields are distinct enough and of similar strength. In the context of a strong or superficial source, weak or deep sources remain invisible, and nearby sources of similar orientation tend not to be separated but interpreted as one source located roughly in between.

Artifacts↗
PubMed1998

Inhibition of GSK-3beta leading to the loss of phosphorylated MAP-1B is an early event in axonal remodelling induced by WNT-7a or lithium.

WNT-7a induces axonal spreading and branching in developing cerebellar granule neurons. This effect is mediated through the inhibition of GSK-3beta, a serine/threonine kinase and a component of the WNT pathway. Lithium, an inhibitor of GSK-3beta, mimics WNT-7a in granule cells. Here we examined further the effect of GSK-3beta inhibition on cytoskeletal re-organisation. Lithium induces axonal spreading and increases growth cone area and perimeter. This effect is associated with the absence or reduction of stable microtubules in spread areas. Lithium induces the loss of a phosphorylated form of MAP-1B, a microtubule associated protein involved in axonal outgrowth. Down-regulation of the phosphorylated MAP-1B, MAP-1B-P, from axonal processes occurs before axonal remodelling is evident. In vitro phosphorylation assays show that MAP-1B-P is generated by direct phosphorylation of MAP-1B by GSK-3beta. WNT-7a, like lithium, also leads to loss of MAP-1B-P from spread axons and growth cones. Our data suggest that WNT-7a and lithium induce changes in microtubule dynamics by inhibiting GSK-3beta which in turn lead to changes in the phosphorylation of MAP-1B. These findings suggest a novel role for GSK-3beta and WNTs in axonal remodelling and identify MAP-1B as a new target for GSK-3beta and WNT.

Animals↗
PubMed1985

The cDNA cloning of a 55-kilodalton protein from guinea pig seminal vesicle. Evidence that the protein is the precursor of a 25-kilodalton basic secretory protein.

Guinea pig seminal vesicle epithelium synthesizes and secretes large amounts of four secretory proteins including a basic protein designated SVP-1. The latter migrates as a protein of 25 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When total cellular RNA from seminal vesicle epithelium was translated in vitro by a rabbit reticulocyte system, SVP-1 was not identifiable as a product. In contrast, a major product of the reticulocyte system was a 55-kDa protein which paradoxically could not be identified as one of the four known secretory proteins or as an abundant tissue protein. We isolated a cDNA clone which corresponded to the mRNA which coded for the 55-kDa protein. The mRNA was 1800 bases and of very high abundance; only the transcripts corresponding to the major secretory proteins SVP-3 and SVP-4 were as conspicuous. From these observations, we decided to test whether the 55-kDa protein was related to SVP-1. The 55-kDa protein was found to share properties of SVP-1, including an alkaline pI and selection by antibody against SVP-1. After proteolytic digestion of the 55-kDa protein, one fragment co-migrated with authentic SVP-1. Furthermore, detectable fragments of the 55-kDa protein smaller than SVP-1 all co-migrated with fragments generated from the proteolytic digestion of SVP-1. Finally, we show that the 55-kDa protein could be processed in vitro by seminal vesicle lumenal extracts to yield a 30-kDa protein and a protein the size of native SVP-1. The best explanation of our results is that the 55-kDa protein represents the primary translation product and precursor of SVP-1. Our proteolytic mapping and in vitro processing studies are consistent with the idea that the 55-kDa protein is a tandem repeat of two SVP-1 molecules. We used the cDNA clone to study the expression of the corresponding gene in different states of androgen depletion and repletion.

Animals↗
PubMed1992

Variation in genomic Alu repeat density as a basis for rapid construction of low resolution physical maps of human chromosomes.

Human DNA restriction fragments containing high numbers of Alu repeat sequences can be preferentially detected in the presence of other human DNA restriction fragments in DNA from human: rodent somatic cell hybrids when the DNA is fragmented with enzymes that cleave mammalian DNA infrequently. This ability to lower the observed human DNA complexity allowed us to develop an approach to order rapidly somatic hybrid cell lines retaining overlapping human genomic domains. The ordering process also generates a relative physical map of the human fragments detected with Alu probe DNA. This process can generate physical mapping information for human genomic domains as large as an entire chromosome (100,000 kb). The strategy is demonstrated by ordering Alu-detected NotI fragments in a panel of mouse: human hybrid cells that span the entire long arm of human chromosome 17.

Chromosome Mapping↗
PubMed1990

The human tyrosine kinase gene (FER) maps to chromosome 5 and is deleted in myeloid leukemias with a del(5q).

A novel member of the SRC tyrosine kinase gene family was recently isolated and characterized (Hao et al., 1989). This FES/FPS-related gene, named FER, lacks the transmembrane and extracellular domains which characterize tyrosine kinases with receptor function. Expression of FER in a wide range of cell types indicates a general role in intracellular signalling or differentiation processes. We have now mapped FER to chromosome 5q14----q23 using in situ hybridization techniques and suggest a more precise location within bands 5q21----q22. This region lies adjacent to a complex domain of growth factors and receptors, many involved in regulation of haematopoiesis. FER maps within a critical segment frequently deleted from chromosome 5 in patients with acute myeloid leukemia or myelodysplastic syndromes and was shown to be deleted in two such patients. It also maps close to the familial polyposis coli locus at 5q22.

Chromosome Deletion↗
PubMed1992

Microtubule-associated proteins 1A and LC2. Two proteins encoded in one messenger RNA.

The deduced amino acid sequence for the filamentous microtubule-associated protein (MAP) 1A, thought to be involved in stabilizing the mature neuronal cytoskeleton, has been determined from a series of overlapping cDNA clones. Though previously described as biochemically and immunologically distinct from MAP1B, we now demonstrate that MAP1A is structurally related to MAP1B, a protein associated with neurite outgrowth and process plasticity. The two MAPs exhibit regional amino acid sequence similarities spanning their potential microtubule binding domains placing both into a new MAP family. The cDNA sequence encoding MAP1A was also found to encode one of its associated light chains (LC) called LC2. Both proteins are found on a single mRNA in the same open reading frame and are translated as a pre-MAP1A/LC2-protein. The topological relationship between MAP1A and LC2 coding sequences is, therefore, identical to that previously shown for MAP1B and LC1 (Hammarback, J. A., Obar, R. A., Hughes, S. M., and Vallee, R. B. (1991) Neuron 7, 129-139). Based on these and earlier results, we conclude that LC1 and LC2 are structurally related polypeptides generated from distinct MAP polyprotein precursors but free to exchange between the two MAPs.

Amino Acid Sequence↗
PubMed1998

The continuous performance test revisited with neuroelectric mapping: impaired orienting in children with attention deficits.

A total of 11 children with attention deficit disorder (ADD) and nine control children performed a continuous performance test (CPT) of the A-X type with concurrent neuroelectric brain mapping to assess preparatory processing, purportedly mediated by the frontal lobes. This cued CPT task proved to be a highly specific task. The groups could be clearly differentiated both at the behavioral and electrophysiological level. ADD children detected fewer signals and made more false alarms. There were no major group differences in topographical distribution of the event-related potential microstates, but ADD children displayed reduced global field power (GFP) in an early CNV/P3 microstate to cues. This indicated that impaired orienting to cues, rather than impaired executive target processing, determines the initial processing stages in ADD. In comparison with data from the same task run in Utrecht, the same orienting deficit in clinically diagnosed ADHD children was demonstrated. Low resolution electromagnetic tomography (LORETA) estimated posterior sources underlying these orienting processes and the orienting deficit. This argued against frontal lobe involvement at this stage and suggested involvement of a posterior attention system.

Arousal↗
PubMed2006

The enteropathogenic Escherichia coli EspF effector molecule inhibits PI-3 kinase-mediated uptake independently of mitochondrial targeting.

Delivery of effector molecules into LMme(v) macrophages by enteropathogenic Escherichia coli, via its type three secretion system (T3SS), inhibits bacterial uptake by a phosphatidylinositol-3 (PI-3) kinase-dependent pathway. The T3SS system, encoded by the locus of enterocyte effacement (LEE) pathogenicity island, delivers LEE- and non-LEE-encoded effector proteins into host cells. Previous studies discounted essential roles for the LEE-encoded Map, EspF, Tir or Intimin proteins in this process but correlated it with loss of phosphorylation of the PI-3 kinase substrate, Akt (Celli et al., 2001, EMBO J 20: 1245-1258). Given the more recent finding that these bacterial proteins are multifunctional and can act together to subvert host cellular processes, we generated a quadruple deletion mutant (Map, Tir, EspF and Intimin deficient) to unearth any cooperativity in inhibiting uptake. The quadruple mutant was as defective as the T3SS-defective strain at preventing bacterial uptake with further studies revealing a surprising dependence on EspF but not Map, Tir or Intimin. Subversive activities previously associated with EspF are disruption of epithelial barrier function and programmed cell death, with the latter linked to EspF targeting mitochondria. Interestingly, the C-terminal domain possesses a polyproline motif associated with protein-protein interactions. We demonstrate that EspF-mediated inhibition of PI-3 kinase-dependent uptake: (i) is independent of mitochondrial targeting, (ii) requires the N-terminal domain with and (iii) the C-terminal domain is sufficient to disrupt barrier function but not inhibition of bacterial uptake. Moreover, loss of PI-3 kinase-dependent phosphorylation of Akt and gross changes in host phosphotyrosine protein profiles could not be linked to inhibition of the PI-3 kinase-dependent uptake process.

Adhesins, Bacterial↗
PubMed2002

A space for measuring mind and brain: interdisciplinarity and digital tools in the development of brain mapping and functional imaging, 1980-1990.

Brain mapping is said to have opened up the possibility of a new collaboration between the sciences of mind and the sciences of the brain, potentially leading to a new kind of scientist, sometimes called "cognitive neuroscientist." This article traces the recent history of brain mapping and analyzes the processes that have led to a new "close working relationship" between the sciences of mind and brain. A key part of the working relationship is shown to be constituted through the development of the Talairach system, a digital space in which to measure structure and function. The development of meaningful brain mapping data involves the creation of measurement spaces that allow interdisciplinary collaboration and is not the result solely of theoretical developments or of the application of a technology.

Brain↗