Ankyrin facilitates intracellular trafficking of alpha1-Na+-K+-ATPase in polarized cells.

Defects in ankyrin underlie many hereditary disorders involving the mislocalization of membrane proteins. Such phenotypes are usually attributed to ankyrin's role in stabilizing a plasma membrane scaffold, but this assumption may not be accurate. We found in Madin-Darby canine kidney cells and in other cultured cells that the 25-residue ankyrin-binding sequence of alpha(1)-Na(+)-K(+)-ATPase facilitates the entry of alpha(1),beta(1)-Na(+)-K(+)-ATPase into the secretory pathway and that replacement of the cytoplasmic domain of vesicular stomatitis virus G protein (VSV-G) with this ankyrin-binding sequence bestows ankyrin dependency on the endoplasmic reticulum (ER) to Golgi trafficking of VSV-G.

Female ROMK null mice manifest more severe Bartter II phenotype on renal function and higher PGE2 production.

ROMK null mice with a high survival rate and varying severity of hydronephrosis provide a good model to study type II Bartter syndrome pathophysiology (26). During the development of such a colony, we found that more male than female null mice survived, 58.7% vs.

The COOH terminus of megalin regulates gene expression in opossum kidney proximal tubule cells.

We recently reported that megalin is subjected to regulated intramembrane proteolysis (RIP) and includes 1) protein kinase C (PKC)-regulated, metalloprotease-mediated ectodomain shedding producing a membrane-bound megalin COOH-terminal fragment (MCTF) and 2) gamma-secretase-mediated cleavage of the MCTF producing a soluble megalin intracellular domain (MICD). Based on studies of RIP of other receptors, the MICD is predicted to target to the nucleus and regulate gene expression. To determine whether RIP of megalin regulates proximal tubule gene expression, we stably expressed the transfected MCTF (tMCTF) or transfected MICD (tMICD) in opossum kidney proximal tubule (OKP) cells and examined the resulting phenotype.

Area and length minimizing flows for shape segmentation.

A number of active contour models have been proposed that unify the curve evolution framework with classical energy minimization techniques for segmentation, such as snakes. The essential idea is to evolve a curve (in two dimensions) or a surface (in three dimensions) under constraints from image forces so that it clings to features of interest in an intensity image. The evolution equation has been derived from first principles as the gradient flow that minimizes a modified length functional, tailored to features such as edges.

Consistency and stability of active contours with Euclidean and non-Euclidean arc lengths.

External energies of active contours are often formulated as Euclidean arc length integrals. In this paper, we show that such formulations are biased. By this we mean that the minimum of the external energy does not occur at an image edge.

Competition and cooperation in neuronal processing.

A new type of model neuron is introduced as a building block of an associative memory. The neuron, which has a number of receptor zones, processes both the amplitude and the frequency of input signals, associating a small number of features encoded by those signals. Using this two-parameter input in our model compared to the one-dimensional inputs of conventional model neurons (e.

A feature-based approach to combine functional MRI, structural MRI and EEG brain imaging data.

The acquisition of multiple brain imaging types for a given study is a very common practice. However these data are typically examined in separate analyses, rather than in a combined model. We propose a novel methodology to perform joint independent component analysis across image modalities, including structural MRI data, functional MRI activation data and EEG data, and to visualize the results via a joint histogram visualization technique.

Differential regulation of cholera toxin-inhibited Na-H exchange isoforms by butyrate in rat ileum.

Electroneutral Na absorption occurs in the intestine via sodium-hydrogen exchanger (NHE) isoforms NHE2 and NHE3. Bicarbonate and butyrate both stimulate electroneutral Na absorption through NHE. Bicarbonate- but not butyrate-dependent Na absorption is inhibited by cholera toxin (CT).

Nuclear receptors RXRalpha:RARalpha are repressors for human MRP3 expression.

Multidrug resistance-associated protein MRP3/Mrp3 (ABCC3) is upregulated in cholestasis, an adaptive response that may protect the liver from accumulation of toxic compounds, such as bile salts and bilirubin conjugates. However, the mechanism of this upregulation is poorly understood. We and others have previously reported that fetoprotein transcription factor/liver receptor homolog-1 is an activator of MRP3/Mrp3 expression.

Adenosine inhibits cytosolic calcium signals and chemotaxis in hepatic stellate cells.

Adenosine is produced during cellular hypoxia and apoptosis, resulting in elevated tissue levels at sites of injury. Adenosine is also known to regulate a number of cellular responses to injury, but its role in hepatic stellate cell (HSC) biology and liver fibrosis is poorly understood. We tested the effect of adenosine on the cytosolic Ca2+ concentration, chemotaxis, and upregulation of activation markers in HSCs.

Role of a tyrosine kinase in the CO2-induced stimulation of HCO3- reabsorption by rabbit S2 proximal tubules.

A previous study demonstrated that proximal tubule cells regulate HCO(3)(-) reabsorption by sensing acute changes in basolateral CO(2) concentration, suggesting that there is some sort of CO(2) sensor at or near the basolateral membrane (Zhou Y, Zhao J, Bouyer P, and Boron WF Proc Natl Acad Sci USA 102: 3875-3880, 2005). Here, we hypothesized that an early element in the CO(2) signal-transduction cascade might be either a receptor tyrosine kinase (RTK) or a receptor-associated (or soluble) tyrosine kinase (sTK). In our experiments, we found, first, that basolateral 17.

Activation of AMPK alpha- and gamma-isoform complexes in the intact ischemic rat heart.

AMP-activated protein kinase (AMPK) plays a key role in modulating cellular metabolic processes. AMPK, a serine-threonine kinase, is a heterotrimeric complex of catalytic alpha-subunits and regulatory beta- and gamma-subunits with multiple isoforms. Mutations in the cardiac gamma(2)-isoform have been associated with hypertrophic cardiomyopathy and pre-excitation syndromes.

Mercury and zinc differentially inhibit shark and human CFTR orthologues: involvement of shark cysteine 102.

The apical membrane is an important site of mercury toxicity in shark rectal gland tubular cells. We compared the effects of mercury and other thiol-reacting agents on shark CFTR (sCFTR) and human CFTR (hCFTR) chloride channels using two-electrode voltage clamping of cRNA microinjected Xenopus laevis oocytes. Chloride conductance was stimulated by perfusing with 10 microM forskolin (FOR) and 1 mM IBMX, and then thio-reactive species were added.

Rapid dilation of arterioles with single contraction of hamster skeletal muscle.

Skeletal muscle blood flow increases rapidly with exercise onset, but little is known of where or how the rapid onset of vasodilation (ROV) is governed within the microcirculation. In the retractor muscle of anesthetized hamsters (n = 26), we tested the following: 1) where in the resistance network ROV occurred, 2) how microvascular responses were affected by the duration of contraction, and 3) whether ROV involved muscarinic receptor activation. Single tetanic contractions were evoked using supramaximal field stimulation (100 Hz) to depolarize motor end plates.

Temporal variability in physical speciation of metals during a winter rain-on-snow event.

Particulate matter in urban rivers transports a significant fraction of pollutants, changes rapidly during storm events, and is difficult to characterize. In this study, the physical speciation of trace metals and organic C in an urban river and upstream headwaters site in Torrington, CT, were measured during a winter rain-on-snow event. In addition, a selective fractionation scheme, using membrane and tangential-flow ultrafiltration methods to separate suspended particulate matter into sand, silt, clay, and colloid fractions, was evaluated based on the appropriateness of the chosen size categories.

Role of short-chain fatty acids in colonic HCO(3) secretion.

Luminal isobutyrate, a relatively poor metabolized short-chain fatty acid (SCFA), induces HCO(3) secretion via a Cl-independent, DIDS-insensitive, carrier-mediated process as well as inhibiting both Cl-dependent and cAMP-induced HCO(3) secretion. The mechanism(s) responsible for these processes have not been well characterized. HCO(3) secretion was measured in isolated colonic mucosa mounted in Lucite chambers using pH stat technique and during microperfusion of isolated colonic crypts.

The COOH-terminal tail of the GAT-2 GABA transporter contains a novel motif that plays a role in basolateral targeting.

The ability of polarized epithelia to perform vectorial transport depends on the asymmetrical distribution of transmembrane proteins among their plasma membrane domains. The establishment and maintenance of these polar distributions relies on molecular signals embedded in the proteins themselves and the interpretation of these signals by cellular sorting machinery. Using Madin-Darby canine kidney (MDCK) cells as an in vitro model of polarized epithelia, our laboratory has previously shown that the COOH-terminal cytoplasmic 22 amino acids of the GAT-2 isoform of the gamma-amino butyric acid (GABA) transporter are necessary for its basolateral distribution.

Expression of P2Y nucleotide receptors and ectonucleotidases in quiescent and activated rat hepatic stellate cells.

Extracellular nucleotides regulate a variety of cellular activities, including proliferation of fibrogenic cells outside of the liver. However, the expression of receptors for extracellular nucleotides in hepatic stellate cells (HSC) is unknown. Thus our aims were to investigate the expression of mediators of nucleotide signaling in HSC and to determine whether extracellular nucleotides regulate HSC function.

Adaptive control using neural networks and approximate models.

The NARMA model is an exact representation of the input-output behavior of finite-dimensional nonlinear discrete-time dynamical systems in a neighborhood of the equilibrium state. However, it is not convenient for purposes of adaptive control using neural networks due to its nonlinear dependence on the control input. Hence, quite often, approximate methods are used for realizing the neural controllers to overcome computational complexity.

Unique presentation of Crohn's disease during treatment of Hodgkin's disease.

Although Hodgkin's disease and Crohn's disease are associated with abnormalities in cell-mediated immunity, their co-existence in an individual patient is uncommon. To our knowledge, this is the first report of the clinical presentation of Crohn's disease during treatment for Hodgkin's disease. The basic immunologic mechanisms underlying these two pathologic states as well as chemotherapy-related effects are postulated as potential etiologic mechanisms for the codevelopment of these diseases.

A Lagrangian relaxation network for graph matching.

A Lagrangian relaxation network for graph matching is presented. The problem is formulated as follows: given graphs G and g, find a permutation matrix M that brings the two sets of vertices into correspondence. Permutation matrix constraints are formulated in the framework of deterministic annealing.

Deformable boundary finding in medical images by integrating gradient and region information.

Accurately segmenting and quantifying structures is a key issue in biomedical image analysis. The two conventional methods of image segmentation, region-based segmentation, and boundary finding, often suffer from a variety of limitations. Here the authors propose a method which endeavors to integrate the two approaches in an effort to form a unified approach that is robust to noise and poor initialization.