Medical Physiology Commons^™

Na/K-Atpase Amplification Of Oxidant Stress; A Universal But Unrecognized Clinical Target?, Zijian Xie, Phd, Joseph I. Shapiro, Md

Joseph I Shapiro MD

The Na/K-ATPase has a signaling function which appears to be separate from its ion pumping function. This signaling function refers to the transduction of conformational changes in the Na/K-ATPase alpha1 subunit into activating Src’s tyrosine kinase activity, triggering a cascade which generates reactive oxygen species (ROS), modulates other signaling pathways, and causes many physiological and pathophysiological effects. We have recently observed that ROS themselves as well as cardiotonic steroids can actually initiate the signal by directly inducing conformational changes in alpha1. It therefore appears that the Na/K-ATPase signal cascade can serve as a feed forward amplification for ROS with circulating …

Effects Of Uremic Serum On Isolated Cardiac Myocyte Calcium Cycling And Contractile Function, Sankaridrug Periyasamy, Jie Chen, Derek Cooney, Patricia Carter, Eiad Omran, Jiang Tian, Snigdha Priyadarshi, Alexei Bagrov, Olga Fedorova, Deepak Malhotra, Zijian Xie, Joseph Shapiro

Zijian Xie

Background: Diastolic dysfunction occurs in patients with chronic renal failure. Moreover, serum from uremic patients contains one or more inhibitors of the plasmalemmal Na,K-ATPase (sodium pump). We hypothesized that a circulating substance present in uremic sera contributes to both sodium pump inhibition and diastolic dysfunction.

Methods: Serum samples were obtained from six patients with chronic renal failure and diastolic dysfunction.

Results: Their serum samples caused marked inhibition of Na,K-ATPase purified from dog kidney at all concentrations studied (all P < 0.01) and also impaired ouabain-sensitive rubidium uptake by myocytes isolated from Sprague-Dawley rats (P < 0.01). These cardiac myocytes were studied for their contractile function with video-edge detection and calcium metabolism with indo-1 fluorescence spectroscopy after exposure to these uremic sera. These uremic sera caused increases in myocyte fractional shortening (P < 0.01) as well as an increase in the time constant of relengthening (P < 0.01). Examining the calcium transient, the time constant for calcium recovery was also increased (P < 0.01). Exposure of these cells to sera from age- and sex-matched healthy subjects did not result in significant changes in contraction or calcium cycling. Extracts of uremic serum samples inhibited isolated Na,K-ATPase whereas extracts of normal serum samples did not. The effect of uremic serum extracts on contractile function and calcium cycling were quite similar to that of intact serum or the addition of ouabain. Co-incubation of uremic serum extract with an antibody fragment directed against digoxin markedly attenuated the inhibition of Na,K-ATPase activity and completely prevented any effects on calcium cycling or contractile function.

Conclusion: These data show that one or more substances are present in uremic sera that acutely cause increased force of contraction …

Cardiac Performance In Inbred Rat Genetic Models Of Low And High Running Capacity, J. Chen, G. Feller, J. Barbato, S. Periyasamy, Zijian Xie, L. Koch, Joseph Shapiro, S. Britton

Zijian Xie

Previous work demonstrating that DA inbred rats are superior to COP inbred rats in aerobic treadmill running capacity has indicated their utility as genetic models to explore this trait. We tested the general hypothesis that intermediate phenotypes of cardiac function and calcium metabolism are responsible for the difference in capacity between these strains.

Logical cardiac trait differences were estimated at a tissue (isolated papillary muscle), cellular (isolated left ventricular cells), and biochemical level of organization.

DA hearts were found to give significantly higher values than COP hearts for: (1) maximal developed tension (38.3 % greater), and rates of tension change …

Extracellular Ubiquitin: Role In Cardiac Myocyte Apoptosis And Myocardial Remodeling, Christopher Daniels

Dr. Christopher C Daniels

Activation of sympathetic nervous system is a key component of myocardial remodeling that generally occurs following ischemia/reperfusion (I/R) injury and myocardial infarction. It induces cardiac myocyte apoptosis and myocardial fibrosis, leading to myocardial dysfunction. Intracellular ubiquitin (UB) regulates protein turnover by the UB-proteosome pathway. The biological functions of extracellular UB in the heart remain largely unexplored. Previously, our lab has shown that β-adrenergic receptor (β-AR) stimulation increases extracellular UB levels, and extracellular UB inhibits β-AR-stimulated apoptosis in adult rat ventricular myocytes (ARVMs). This study explores the role of extracellular UB in myocyte apoptosis, fibroblast phenotype and function, and myocardial remodeling …

A Comparison Of Four Electrical Stimulation Types On Staphylococcus Aureus Growth In Vitro, Harold Merriman, Chris Hegyi, Cheryl Albright-Overton, John Carlos, Robert Putnam, Janet Mulcare

Harold L. Merriman

We evaluated the efficacy of common electrical stimulation (ES) types on bacterial growth in vitro using clinically relevant conditions. Four types of ES-continuous microamperage direct current (µADC), high-voltage pulsed current (HVPC), low-voltage monophasic milliamperage pulsed current (LVMmAPC), and low-voltage biphasic milliamperage pulsed current (LVBmAPC)-were each applied to a separate set of culture plates containing Staphylococcus aureus for 1 h at 37 °C on 3 consecutive days. After ES treatment, the zone of inhibition surrounding each electrode was measured. Zone of inhibition measurements showed a significant inhibitory effect for continuous µADC and HVPC (p < 0.05), but not for LVMmAPC and LVBmAPC. …

Características Sociodemográficas E Aspectos Clínicos De Pacientes Com Doença Renal Policística Do Adulto Submetidos À Hemodiálise; Sociodemographic Characteristics And Clinical Features Of Patients With Adult Polycystic Kidney Disease Undergoing Hemodialysis, Everton Alves, Luiza Tsuneto, Sueli Borelli, Renata Cadidé, Rosane De Freitas, Ângela Gravena, Sandra Pelloso, Maria Dalva Carvalho

Everton Fernando Alves

Aims: To analyze the socio-demographic and clinical characteristics of patients with adult polycystic kidney disease

admitted to hemodialysis services in Northwestern Paraná state, Brazil.

Methods: This was an observational, descriptive and retrospective longitudinal study. Medical records of patients

with polycystic kidneys who initiated hemodialysis between 1995 and 2012, in four centers that treat patients of the

coverage area of the 15th Regional Health Region of Paraná state where analyzed.

Results: We found that 10.3% of hemodialysis patients had polycystic kidney disease as a leading cause of stage 5 of

chronic kidney disease. The mean age of patients was 54.9±9.4 years …

Effects Of Dietary Magnesium On Testicular Histology, Steroidogenesis, Spermatogenesis And Oxidative Stress Markers In Adult Rats, Amar K. Chandra, Pallav Sengupta, Haimanti Goswami, Mahitosh Sarkar

Pallav Sengupta, PhD

Regulated Proteolysis Controls Mucoid Conversion In Pseudomonas Aeruginosa, Dongru Qiu, Vonya Eisinger, Donald Rowen, Hongwei Yu

Hongwei Yu

Overproduction of the exopolysaccharide alginate causes mucoid conversion in Pseudomonas aeruginosa and is a poor prognosticator in cystic fibrosis. The ECF σ factor AlgU and its cognate anti-σ factor MucA are two principal regulators of alginate production. Here, we report the identification of three positive regulators of alginate biosynthesis: PA4033 (designated mucE), PA3649 (designated mucP), and algW. MucE, a small protein (9.5 kDa), was identified as part of a global mariner transposon screen for new regulators of alginate production. A transposon located in its promoter caused the overexpression of MucE and mucoid conversion in P. aeruginosa strains PAO1 and PA14. …

A Dominant Loss-Of-Function Gja1 (Cx43) Mutant Impairs Parturition In The Mouse, Dan Tong, Xuerong Lu, Hong-Xing Wang, Isabelle Plante, Ed Lui, Dale Laird, Donglin Bai, Gerald Kidder

Edmund M. K. Lui

Expression of GJA1 (commonly known as connexin43 or Cx43), a major myometrial gap junction protein, is upregulated before the onset of delivery, suggesting an essential role for Cx43-mediated gap junctional intercellular communication (GJIC) in normal uterine contraction during parturition. To determine how a disease-linked Cx43 mutation affects myometrial function, we studied a mutant mouse model carrying an autosomal dominant mutation (Gja1Jrt) in the gene encoding Cx43 that displays features of the human genetic disease oculodentodigital dysplasia. We found that Cx43 level, specifically the phosphorylated species of the protein, is significantly reduced in the myometrium of the mutant mice (Gja1Jrt/+), as …

Analysis Of Oocyte-Like Cells Differentiated From Porcine Fetal Skin-Derived Stem Cells, Paul Dyce, Wei Shen, Evanna Huynh, Hua Shao, Daniel Villagómez, Gerald Kidder, W Allan King, Julang Li

Paul W. Dyce

We previously reported the differentiation of cells derived from porcine female fetal skin into cells resembling germ cells and oocytes. A subpopulation of these cells expressed germ cell markers and formed aggregates resembling cumulus-oocyte complexes. Some of these aggregates extruded large oocyte-like cells (OLCs) that expressed markers consistent with those of oocytes. The objective of the current study was to further characterize OLCs differentiated from porcine skin-derived stem cells. Reverse transcriptase (RT)-polymerase chain reaction and Western blot revealed the expression of connexin37 and connexin43, both of which are characteristic of ovarian follicles. The expression of meiosis markers DMC1 and synaptonemal …

Learning From Mistakes: Improving Initial Fingertip Force Scaling By Observing Lifting Errors, Gavin Buckingham, Minnie Tang, Paul Gribble, Melvyn Goodale

Gavin Buckingham

• When lifting objects that are lighter or heaver than we expect them to be, individuals typically misapply forces in a way that reflects their prior expectations of heaviness.

• Because we lift in this predictive way, large and small cubes elicit these characteristic errors even when they are adjusted to have equal mass. Lifters will apply too much force to a large cube and substantially less force to a small cube – errors that are rapidly corrected with repeated lifts (Flanagan & Beltzner, 2000).

• When watching others lift objects, an observer’s motor system automatically reacts in a way …

Pharmacologic Responses Of The Mouse Urinary Bladder, A. Erdem, Christopher Chapple, Russ Chess-Williams

Russ Chess-Williams

The aim of the study was to determine pathways involved in contraction and relaxation of the mouse urinary bladder. Mouse bladder strips were set up in gassed Krebs-bicarbonate solution and responses to various drugs and electrical field stimulation were obtained. Isoprenaline (b-receptor agonist) caused a 63% inhibition of carbachol precontracted detrusor (EC50=2nM). Carbachol caused contraction (EC50=0.3µM), responses were antagonised more potently by 4-DAMP (M3-antagonist) than methoctramine (M2-antagonist). Electrical field stimulation caused contraction, which was inhibited by atropine (60%) and less by guanethidine and α,β-methylene-ATP. The neurogenic responses were not potentiated by inhibition of nitric oxide synthase. Presence of an intact …

Pharmacologic Responses Of The Mouse Urinary Bladder, A. Erdem, Christopher Chapple, Russ Chess-Williams

Russ Chess-Williams

The aim of the study was to determine pathways involved in contraction and relaxation of the mouse urinary bladder. Mouse bladder strips were set up in gassed Krebs-bicarbonate solution and responses to various drugs and electrical field stimulation were obtained. Isoprenaline (b-receptor agonist) caused a 63% inhibition of carbachol precontracted detrusor (EC50=2nM). Carbachol caused contraction (EC50=0.3µM), responses were antagonised more potently by 4-DAMP (M3-antagonist) than methoctramine (M2-antagonist). Electrical field stimulation caused contraction, which was inhibited by atropine (60%) and less by guanethidine and α,β-methylene-ATP. The neurogenic responses were not potentiated by inhibition of nitric oxide synthase. Presence of an intact …

De Novo Sequencing And Analysis Of The American Ginseng Root Transcriptome Using A Gs Flx Titanium Platform To Discover Putative Genes Involved In Ginsenoside Biosynthesis, Chao Sun, Ying Li, Qiong Wu, Hongmei Luo, Yongzhen Sun, Jingyuan Song, Ed Lui, Shilin Chen

Edmund M. K. Lui

BACKGROUND: American ginseng (Panax quinquefolius L.) is one of the most widely used herbal remedies in the world. Its major bioactive constituents are the triterpene saponins known as ginsenosides. However, little is known about ginsenoside biosynthesis in American ginseng, especially the late steps of the pathway. RESULTS: In this study, a one-quarter 454 sequencing run produced 209,747 high-quality reads with an average sequence length of 427 bases. De novo assembly generated 31,088 unique sequences containing 16,592 contigs and 14,496 singletons. About 93.1% of the high-quality reads were assembled into contigs with an average 8-fold coverage. A total of 21,684 (69.8%) …

Lifting Without Seeing: The Role Of Vision In Perceiving And Acting Upon The Size‐Weight Illusion, Gavin Buckingham, Melvyn Goodale

Gavin Buckingham

Our expectations of an object’s heaviness not only drive our fingertip forces, but also our perception of heaviness. This effect is highlighted by the classic size-weight illusion (SWI), where different‐sized objects of identical mass feel different weights (Charpentier, 1891) long after any initial errors in the application of fingertip forces have been corrected (Flanagan & Beltzner, 2000).

Here, we examined whether our expectations about the weight of an upcoming lift are sufficient to induce the SWI in a single wooden cube when lifted without visual feedback, by varying the size of the object seen prior to the lift during a …

Laterality, Perception, And Action During The Size-Weight Illusion, Gavin Buckingham, Nathalie Ranger, Melvyn Goodale

Gavin Buckingham

In the classic size-weight illusion (SWI), a small object will feel heavier than an larger object of equal weight (Charpentier, 1891). Individuals continue to perceive this illusory difference in weight long after their gripping and lifting forces have scaled to the actual, identical, mass of the illusion-inducing stimuli (Flanagan & Beltzner, 2000).

The independence of our weight perception and fingertip force application has only been quantified in the right hand of right-handers. The immunity to this perceptual illusion may be affected by manual asymmetries (e.g., Gonzalez, Ganel & Goodale, 2006).

We examined perception of heaviness and fingertip force scaling in …

Effects Of Estrogen On The Mechanical Behavior Of The Human Achilles Tendon In Vivo, Adam Bryant, Ross Clark, Simon Bartold, Aron Murphy, Kim Bennell, Erik Hohmann, Sonya Marshall-Gradisnik, Craig Payne, Kay Crossley

Sonya Marshall-Gradisnik

The purpose of this study was to elucidate the effect of normal fluctuating [nonmonophasic oral contraceptive pill (MOCP) users] and low, consistent (MOCP users) endogenous plasma estrogen levels on the strain behavior of the Achilles tendon in vivo. Twenty women (age 28.0 ± 4.2 yr, height 1.67 ± 0.07 m, mass 61.6 ± 6.8 kg) who had been using the MOCP for at least 12 mo together with 20 matched women who were non-MOCP users (age 31.9±7.3 yr, height 1.63 ± 0.05 m, mass 62.5 ± 5.9 kg) participated in this study. Non-MOCP users were tested at the time of …

A Dissociation Between Perception And Action In The Material‐Weight Illusion, Gavin Buckingham, Jonathan Cant, Kai-Ling Kao, Melvyn Goodale

Gavin Buckingham

We examined what forces are applied to objects that elicit this illusion when they are lifted.We predicted that:

(1) Forces on early trials will scale to each participant’s expectations of how much a particular block will weigh ‐ excessive force will be applied to the metal block and insufficient force applied to the polystyrene block.

(2) Forces on later trials will scale to the real weight of each block ‐ identical levels of force applied to all the blocks.

(3) MWI will persist throughout ‐ polystyrene block will feel the heaviest, metal block will feel the lightest.

Grasping And Lifting Different Materials, Gavin Buckingham, Jonathan Cant, Melvyn Goodale

Gavin Buckingham

The material from which an object is made can determine how heavy it feels (Seashore, 1899). Interestingly, a metal block that has been adjusted to have the same size and mass as a polystyrene block will feel lighter than the polystyrene block. We recently showed that participants experiencing this material-weight illusion’ (MWI) do not apply forces that match their perceptual experience of heaviness ‐ just like in the size‐weight illusion ( Flanagan & Beltzner, 2000).

Our previous study showed that forces on early trials were scaled to each participant’s expectations of how much a particular block should weigh ‐ excessive …

Exaggerated Natriuretic Response Of Brattleboro Rats To Extracellular Volume Expansion, Susan C. Opava-Stitzer, Emma Fernandez-Repollet, Carmen Rodriguez-Sargent, Jose L. Cangiano, Manuel Martinez-Maldonado

Susan C. Opava-Stitzer

A state of chronic dehydration with reduced plasma volume, decreased blood pressure, and increased plasma renin activity (PRA) has been demonstrated in rats with hereditary hypothalamic diabetes insipidus (DI) rats. In this situation decreased renal perfusion and glomerular filtration rate might result in sodium retention. On the other hand, the DI rat also suffers from mineralocorticoid deficiency which might result in salt wasting. In addition it has recently been shown that in contrast to normal rats, there are no differences between superficial cortical and juxtamedullary nephrons of the DI rat with respect to single nephron filtration rate, glomerular volume, and …

Effect Of Potassium On Plasma Renin Concentration In The Presence And Absence Of Adh (Brattleboro Rat Model), Emma Fernandez-Repollet, Susan C. Opava-Stitzer, Manuel Martinez-Maldonado

Susan C. Opava-Stitzer

Rats with hereditary hypothalamic diabetes insipidus (so-called DI rats) have elevated plasma renin levels. Although the mechanism responsible for this condition has not be elucidated, it seems reasonable to postulate that the absence of ADH and/or the hypokalemia previously reported in these rats might contribute to the elevation of plasma renin concentration (PRC). Evidence in favor of this hypothesis emerges from studies in which both ADH and potassium have been shown to inhibit renin release. In an attempt to examine the relative roles of ADH and potassium in the regulation of renin secretion, PRC was measured in DI rats maintained …

Effect Of Lithium And Antidiuretic Hormone On Plasma Renin Concentration In Diabetes Insipidus Rats (Brattleboro Rat Model), Susan C. Opava-Stitzer

Susan C. Opava-Stitzer

Antidiuretic horomone (ADH) is known to inhibit renin secretion in many species, but the mechanism of this inhibition and its importance in the control of renin secretion are unknown.

Evidence For Baroreceptor Control Of Renin Release, Paul C. Churchill, Richard L. Malvin, Susan C. Opava-Stitzer

Susan C. Opava-Stitzer

Arterial plasma renin activity (PRA) was measured using radioimmunoassay techniques in pentobarbital-anesthetized dogs. Acute saline loading (40-60ml 150mm NaCl/kg body weight, or 20ml 300mm NaCl/kg body weight, i.v.) reduced PRA from control level in 7 dogs. Despite continued increases in filtered and excreted masses of Na, subsequent partial aortic clamping increased PRA in proportion to the degree of reduction in renal perfusion pressure. In 6 other dogs aortic clamping super-imposed on total ureteral occlusion led to changes in PRA which were inversely associated with renal perfusion pressure. These results suggest a controlling mechanism for renin secretion sensitive to some hemodynamic …

Urine Concentration And Dilution In The Rat: Contribution Of Papillary Structures During High Rates Of Urine Flow, Manuel Martínez-Maldonado, Susan C. Opava-Stitzer

Susan C. Opava-Stitzer

To examine the contribution of papillary structures to the overall process of urine dilution and concentration at high rates of flow, studies were performed in unilaterally papillectomized kidneys (PX). Comparison of kidney weights in sham-operated and PX rats revealed a significant reduction in total weight of the latter. Papillary length removed was 3045 ± 423 μm. GFR was reduced by 24% and 45% in sham and PX kidneys when compared to their contralateral controls. Under hydropenic conditions, maximal urine concentrating ability (Umax) was comparable in control and sham kidneys but was 59% less in PX kidneys. Fractional sodium excretion during …

Role Of Water Balance In The Enhanced Potassium Excretion And Hypokalaemia Of Rats With Diabetes Insipidus, Emma Fernandez-Repollet, Manuel Martínez-Maldonado, Susan C. Opava-Stitzer

Susan C. Opava-Stitzer

1. The role of water balance in the hypokalaemia of rats with diabetes insipidus (DI rats) was studied. 2. After a 3-day balance study DI rats had a lower muscle potassium content, and plasma [K+], and the urinary excretion of potassium in response to oral KCl loading was reduced when compared to normal rats. The hypokalaemia was found to be associated with elevated concentrations of potassium in renal medulla and papilla when compared to values in normal Long-Evans rats. 3. During a 9-day balance study urinary potassium excretion was higher than that of normal rats on days 1-3, but not …

Pathophysiology Of Clinical Disorders Of Urine Concentration And Dilution, Manuel Martínez-Maldonado, Susan C. Opava-Stitzer

Susan C. Opava-Stitzer

A defect in the ability to concentrate or dilute the urine can be easily recognized by the maximum or minimum urine concentration the patient is able to achieve. Maximum concentrating ability (Umax) is determined by the urine osmolality reached after a fixed period of dehydration and maximal diluting ability (Umin) by the minimum osmolality of the urine after the oral ingestion of a fixed water-load. These indices, however, do not allow an understanding of the pathophysiological alterations leading to the presence of the defect.

High-Dose Pralidoxime For Organophosphorus Poisoning, Ashish Goel, Praveen Aggarwal, Sanjeev Bhoi, Vineet Gupta

Vineet Gupta, MD, FACP

No abstract provided.

Antioxidant Effect Of Zinc And Zinc-Metallothionein In The Acute Cytotoxicity Of Hydrogen Peroxide In Ehrlich Ascites Tumour Cells, Zacharias Suntres, Ed Lui

Edmund M. K. Lui

This study was concerned with the role of zinc (Zn) and zinc-metallothionein (Zn-MT) in oxidative stress. Hydrogen peroxide-induced oxidative injury was examined in Ehrlich ascites tumour cells isolated from control host mice, mice pretreated with 10 mg/kg ZnSO4 (i.p.) to increase cellular Zn/Zn-MT levels, and mice exposed to Zn-deficient diet to reduce the cellular Zn/Zn-MT levels. The results of the present study showed that Ehrlich cells with seven-fold differences in Zn-MT concentrations could be obtained by manipulating the Zn status of host mice and that high Zn and Zn-MT levels can make Ehrlich cells more resistant to H2O2-induced oxidative injury …

Biochemical Characterization Of The Major Sorghum Grain Peroxidase, Mamoudou H. Dicko, Harry Gruppen, Riet Hilhorst, Alphons G. J. Voragen, Willen W. H. Van Berkel

Pr. Mamoudou H. DICKO, PhD

Prooxidative Effect Of Copper-Metallothionein In The Acute Cytotoxicity Of Hydrogen Peroxide In Ehrlich Ascites Tumour Cells, Zacharias Suntres, Ed Lui

Edmund M. K. Lui

This study was concerned with the role of copper (Cu) and Cu-metallothionein (Cu-MT) in oxidative stress. Hydrogen peroxide (H(2)O(2))-induced oxidative injury was examined in Ehrlich ascites tumour cells isolated from host mice pretreated with 0, 1 or 2mg of CuSO(4) (ip) 24h earlier. Control Ehrlich cells contained low levels of Cu and Cu treatment produced dose-related increases in cellular Cu and Cu-MT levels and corresponding increases in sensitivity to oxidative toxicity of H(2)O(2) (LC(50), cell blebbing, lipid peroxidation, GSH depletion, and increase in intracellular free [Ca(2+)](i)). Hydrogen peroxide treatment also resulted in the oxidation of MT thiolates, reduction in the …