Search PubMed⌕ Search

Biomedical subjects

S A Lewis

Publications and source records attributed to S A Lewis.

At least 55 records · Page 3Linked to original sources
PubMed2000

Everything you wanted to know about the bladder epithelium but were afraid to ask.

The mammalian urinary bladder epithelium (urothelium) performs the important function of storing urine for extended periods, while maintaining the urine composition similar to that delivered by the kidneys. The urothelium possesses four properties to perform this function. First, it offers a minimum epithelial surface area-to-urine volume; this reduces the surface area for passive movement of substances between lumen and blood. Second, the passive permeability of the apical membrane and tight junctions is very low to electrolytes and nonelectrolytes. Third, the urothelium has a hormonally regulated sodium absorptive system; thus passive movement of sodium from blood to urine is countered by active sodium reabsorption. Last, the permeability properties of the apical membrane and tight junctions of the urothelium are not altered by most substances found in the urine or blood. The importance of the barrier function of the urothelium is illustrated by infectious cystitis. The loss of the barrier function results in the movement of urinary constituents into the lamina propria and underlying muscle layers, resulting in suprapubic and lower back pain and frequent, urgent, and painful voiding.

Animals↗
PubMed2000

Urea modifies the permeability of the mammalian urothelium.

PURPOSE: This study investigated the effects of mucosal (urine side) and serosal (blood side) urea on the permeability properties of the in vitro mammalian urinary bladder epithelium. MATERIALS AND METHODS: The permeability properties of the rabbit urinary bladder epithelium were studied in modified Ussing chambers using electrophysiological techniques. RESULTS: Addition of two molar urea to the mucosal solution did not cause a significant change in the short circuit current (Isc, a measure of the ion transport capacity of the epithelium), or the transepithelial conductance (Gt, a measure of the ability of ions to diffuse across the epithelium). In contrast, addition of 0.5 M urea to the serosal solution caused an increase in Gt of approximately 35 microS/cm.2 as well as an increase in Isc over a 5 minute period. The site of the conductance increase by short-term serosal urea was at the apical membrane and not at the tight junctions. The urea-induced conductance completely reversed upon removal of urea, was non-selective, and the magnitude was voltage dependent. Long term serosal urea (greater than 30 minutes) resulted in an irreversible increase in transepithelial conductance. Mucosal urea altered the time course but not the magnitude of the serosal urea-induced conductance. CONCLUSIONS: The ion permeability of the mammalian urinary bladder is increased by serosal urea. At short times the increase is at the apical membrane, while at long times the increase is at the tight junctions. The presence of mucosal urea slows the loss of urothelial barrier function caused by serosal urea.

Animals↗
PubMed1999

Tubulin folding cofactors as GTPase-activating proteins. GTP hydrolysis and the assembly of the alpha/beta-tubulin heterodimer.

In vivo, many proteins must interact with molecular chaperones to attain their native conformation. In the case of tubulin, newly synthesized alpha- and beta-subunits are partially folded by cytosolic chaperonin, a double-toroidal ATPase with homologs in all kingdoms of life and in most cellular compartments. alpha- and beta-tubulin folding intermediates are then brought together by tubulin-specific chaperone proteins (named cofactors A-E) in a cofactor-containing supercomplex with GTPase activity. Here we show that tubulin subunit exchange can only occur by passage through this supercomplex, thus defining it as a dimer-making machine. We also show that hydrolysis of GTP by beta-tubulin in the supercomplex acts as a switch for the release of native tubulin heterodimer. In this folding reaction and in the related reaction of tubulin-folding cofactors with native tubulin, the cofactors behave as GTPase-activating proteins, stimulating the GTP-binding protein beta-tubulin to hydrolyze its GTP.

Binding Sites↗
PubMed1999

A chaperone with a hydrophilic surface.

The folding of native tubulin involves at least seven different chaperone proteins: prefoldin, the cytosolic chaperonin CCT and five tubulin-specific chaperone proteins named cofactors A-E. The structure of the yeast homolog of cofactor A, Rbl2p, shows it to be a dimer with largely hydrophilic surfaces, reflecting the fact that it interacts with quasi-native, not unfolded, beta-tubulin.

Animals↗
PubMed1999

Eosinophil peroxidase increases membrane permeability in mammalian urinary bladder epithelium.

Eosinophil peroxidase (EPO), a cationic protein found in eosinophils, has been reported to be cytotoxic independent of its peroxidase activity. This study investigated with electrophysiological methods whether EPO is toxic to mammalian urinary bladder epithelium. Results indicate that EPO, when added to the mucosal solution, increases apical membrane conductance of urinary bladder epithelium only when the apical membrane potential is cell interior negative. The EPO-induced conductance was concentration dependent, with a maximum conductance of 411 microseconds/cm2 and a Michaelis-Menten constant of 113 nM. The EPO-induced conductance was nonselective for K+ and Cl-. The conductance was partially reversed using voltage but not by removal of EPO from the bulk solution. Mucosal Ca2+ reversed the EPO-induced conductance by a mechanism involving reversible block of the conductance. Prolonged exposure (up to 1 h) to EPO was toxic to the urinary bladder epithelium, as indicated by an irreversible increase in transepithelial conductance. These results suggest that EPO is indeed toxic to urinary bladder epithelium via a mechanism that involves an increase in membrane permeability.

Animals↗
PubMed1999

Potassium nitrite reaction with 11-nor-delta 9-tetrahydrocannabinol-9-carboxylic acid in urine in relation to the drug screening analysis.

Recently potassium nitrite has been used as an adulterant to interfere with the analysis of 11-nor-delta 9-tetrahydro-cannabinol-9-carboxylic acid (THC-COOH) in urine. A comprehensive study of the THC-COOH and nitrite reaction chemistry and stability under various conditions is presented. Reverse phase high performance liquid chromatography (HPLC) and negative electrospray mass spectrometry (ESMS) results are given to substantiate the derived reaction mechanism and properties leading to reaction termination. The addition of potassium carbonate as a buffering agent prior to or following sample void as a means of preventing the formation of a nitroso-complexed form of the 11-nor-delta 9+-tetrahydrocannabinol-9-carboxylic acid is evaluated.

Carbonates↗
PubMed1998

Prefoldin, a chaperone that delivers unfolded proteins to cytosolic chaperonin.

We describe the discovery of a heterohexameric chaperone protein, prefoldin, based on its ability to capture unfolded actin. Prefoldin binds specifically to cytosolic chaperonin (c-cpn) and transfers target proteins to it. Deletion of the gene encoding a prefoldin subunit in S. cerevisiae results in a phenotype similar to those found when c-cpn is mutated, namely impaired functions of the actin and tubulin-based cytoskeleton. Consistent with prefoldin having a general role in chaperonin-mediated folding, we identify homologs in archaea, which have a class II chaperonin but contain neither actin nor tubulin. We show that by directing target proteins to chaperonin, prefoldin promotes folding in an environment in which there are many competing pathways for nonnative proteins.

Actins↗
PubMed1998

Tumor necrosis factor-alpha increases sodium and chloride conductance across the tight junction of CACO-2 BBE, a human intestinal epithelial cell line.

CACO-2 BBE was used to determine the response of a gastrointestinal epithelium to tumor necrosis factor-alpha (TNF). Incubation of CACO-2 BBE with TNF did not produce any effect on transepithelial resistance (TER) within the first 6 hr but resulted in a 40-50% reduction in TER and a 30% decrease in 1SC (short circuit current) relative to time-matched control at 24 hr. The decrease in TER was sustained up to 1 week following treatment with TNF and was not associated with a significant increase in the transepithelial flux of [14C]-D-mannitol or the penetration of ruthenium red into the lateral intercellular space. Dilution potential and transepithelial 22Na+ flux studies demonstrated that TNF-treatment of CACO-2 BBE cell sheets increased the paracellular permeability of the epithelium to Na+ and Cl-. The increased transepithelial permeability did not associate with an increase in the incidence of apoptosis. However, there was a TNF-dependent increase in [3H]-thymidine labeling that was not accompanied by a change in DNA content of the cell sheet. The increase in transepithelial permeability was concluded to be across the tight junction because: (i) 1 mM apical amiloride reduced the basolateral to apical flux of 22Na+, and (ii) dilution potential studies revealed a bidirectionally increased permeability to both Na+ and Cl-. These data suggest that the increase in transepithelial permeability across TNF-treated CACO-2 BBE cell sheets arises from an alteration in the charge selectivity of the paracellular conductive pathway that is not accompanied by a change in its size selectivity.

Caco-2 Cells↗
PubMed1998

Consistent effects of high socioeconomic status and low birth order, and the modifying effect of maternal smoking on the risk of allergic disease during childhood.

Birth order, maternal age, gestational age, birth weight, maternal smoking, and social class have all been associated with allergic rhinitis, eczema and asthma in childhood, but the consistency of independent effects of these exposures in relation to all of these allergic conditions has not been investigated. We have compared and contrasted the independent effects of these putative risk factors on parent-reported hayfever, eczema and wheeze by age 16 years and in the past 12 months at age 16 using data from the 1970 British birth cohort. The 1970 British birth cohort comprised all children born in England, Scotland and Wales in one week of April 1970, and follow-up surveys at birth, 5, 10 and 16 years of age involved a cumulative total of 17,427 children. We have used data on over 6000 children with complete data at every stage. Social advantage was a risk factor common to each of wheeze at, and hayfever and eczema at and by age 16. Low birth order was an independent risk factor for eczema and hayfever at age 16, but not for wheeze. However, wheeze at age 16 was significantly increased in relation to maternal smoking, with a significant interaction such that the effect of smoking was greatest in those of high birth order and, in the absence of maternal smoking, low birth order was a risk factor for wheeze at age 16. Social advantage and low birth order appear to be the independently consistent determinants of atopic disease. Maternal smoking is an additional risk factor for wheeze, which applies primarily in low socioeconomic groups.

Adolescent↗
PubMed1998

Measles infection, measles vaccination and the effect of birth order in the aetiology of hay fever.

BACKGROUND: It has recently been suggested that measles infection may reduce the risk of atopy. OBJECTIVE: To study the independent effect of measles infection and measles vaccination on the occurrence of hay fever in a British national birth cohort. METHODS: In over 6000 children born in 1970, details of immunizations and childhood diseases were collected by parental interviews at ages 5, 10 and 16 years, and hay fever within the past year at age 16 years. RESULTS: In univariate analysis, hay fever was less common in those contracting measles infection than in those not infected (OR 0.86, 95% CI 0.76-0.96), and more common in those given measles vaccination than in those not vaccinated (OR 1.16, 95% Cl 1.03-1.31). However, these effects were strongly confounded by birth order, which was closely associated with the likelihood of receiving measles vaccination and with the risk of hay fever. A strong interaction between the effects of measles vaccination and infection, and birth order was found, such that in those with many older sibling contacts, hay fever was significantly and independently reduced in relation to both measles infection and measles vaccination relative to those who were neither infected nor vaccinated. CONCLUSIONS: Both measles infection and measles vaccination in childhood appear to reduce the risk of hay fever in children with multiple older sibling contacts. Differential exposure or response to the measles virus may explain the effect of birth order on the occurrence of allergic disease.

Adult↗
PubMed1998

Eosinophil major basic protein increases membrane permeability in mammalian urinary bladder epithelium.

The eosinophil granule protein major basic protein (MBP) is toxic to a wide variety of cell types, by a poorly understood mechanism. To determine whether the action of MBP involves an alteration in membrane permeability, we tested purified MBP on rabbit urinary bladder epithelium using transepithelial voltage-clamp techniques. Addition of nanomolar concentrations of MBP to the mucosal solution caused an increase in apical membrane conductance only when the voltage across the apical membrane was cell interior negative. The magnitude of the MBP-induced conductance was a function of MBP concentration, and the rate of the initial increase in conductance was a function of the transepithelial voltage. The MBP-induced conductance was nonselective for K+ and Cl-. Mucosal Ca2+ reversed the induced conductance, whereas mucosal Mg2+ partially blocked the induced conductance and slowed the rate of the increase in conductance. The induced conductance was partially reversed by changing the voltage gradient across the apical membrane to cell interior positive. Prolonged exposure resulted in an irreversible loss of the barrier function of the urinary bladder epithelium. These results suggest that an increase in cell membrane ion permeability is an initial step in MBP-induced loss of barrier function.

Animals↗
PubMed1998

Modulation of polymyxin B effects on mammalian urinary bladder.

This report demonstrates that Ca2+, Mg2+, and protons alter the ability of polymyxin B (PX, a cationic antibiotic used clinically as a bactericidal agent) to increase the apical membrane conductance of the rabbit urinary bladder. Using electrophysiological methods, we determine that these alterations occur by two mechanisms. First, they blocked the PX-induced conductance in a rapid and reversible manner; second, they competed with PX for a membrane binding site. In addition, Ca2+ (but not Mg2+ or protons) altered the rate at which the induced conductance could be reversed. When solution pH was greater than 8.8, PX was not able to induce a conductance. This ability of high pH to inhibit the action of PX was due to a decrease in the number of positive charges on PX. Further studies demonstrated that for maximal activity, PX required its fatty acid tail. These data were used to develop a model describing the mechanism by which PX can induce a conductance in the apical membrane of the rabbit urinary bladder.

Animals↗
PubMed1998

Tactile illusion-produced number perception in blind and sighted persons.

OBJECTIVE: To determine whether tactile receptive communication is possible in both sighted and blind persons through the use of a mechanical device that creates the perception of continuous contact through sequential points of vibration on the skin and whether the ability of these two groups differs. MATERIAL AND METHODS: The cohort consisted of 20 sighted and 20 blind subjects who had no prior exposure to a tactile communication device that produces a tactile illusion. Each subject was presented number sequences created by the tactile illusion in the form of single-digit, three-digit, and four-digit sequences, and the number of trials needed for correct identification was recorded. RESULTS: On average, the sighted subjects tended to identify the digit sequences sooner than did the blind subjects by almost one trial; however, this difference was not statistically significant. Younger subjects and female subjects also tended to identify the digit sequences sooner. Nevertheless, the overall correlation between age and number of trials across all replications was relatively minor. CONCLUSION: Tactile receptive communication is possible through the use of a mechanical device used to create tactile illusions. The tactile illusion of numbers is equally perceived by blind and sighted persons.

Adult↗
PubMed1997

Tubulin subunits exist in an activated conformational state generated and maintained by protein cofactors.

The production of native alpha/beta tubulin heterodimer in vitro depends on the action of cytosolic chaperonin and several protein cofactors. We previously showed that four such cofactors (termed A, C, D, and E) together with native tubulin act on beta-tubulin folding intermediates generated by the chaperonin to produce polymerizable tubulin heterodimers. However, this set of cofactors generates native heterodimers only very inefficiently from alpha-tubulin folding intermediates produced by the same chaperonin. Here we describe the isolation, characterization, and genetic analysis of a novel tubulin folding cofactor (cofactor B) that greatly enhances the efficiency of alpha-tubulin folding in vitro. This enabled an integrated study of alpha- and beta-tubulin folding: we find that the pathways leading to the formation of native alpha- and beta-tubulin converge in that the folding of the alpha subunit requires the participation of cofactor complexes containing the beta subunit and vice versa. We also show that sequestration of native alpha-or beta-tubulins by complex formation with cofactors results in the destabilization and decay of the remaining free subunit. These data demonstrate that tubulin folding cofactors function by placing and/or maintaining alpha-and beta-tubulin polypeptides in an activated conformational state required for the formation of native alpha/beta heterodimers, and imply that each subunit provides information necessary for the proper folding of the other.

Amino Acid Sequence↗
PubMed1997

Interleukin 10 prevents cytokine-induced disruption of T84 monolayer barrier integrity and limits chloride secretion.

BACKGROUND & AIMS: The proinflammatory cytokine interferon gamma (IFN-gamma) disrupts epithelial barrier integrity and attenuates secretagogue-induced chloride secretion. This study tested the efficacy of the anti-inflammatory cytokine interleukin 10 (IL-10) in maintaining epithelial barrier and chloride secretory function in the presence of IFN-gamma. METHODS: T84 epithelial cell monolayers were treated with IL-10, IFN-gamma, or IFN-gamma plus IL-10. Monolayer barrier integrity was assessed by measurements of electrical conductance, unidirectional mannitol and inulin fluxes, and tight junctional charge selectivity in Ussing chambers. Short-circuit current (Isc) was measured in response to carbachol and forskolin stimulation. RESULTS: IL-10 attenuated the IFN-gamma-induced increase in electrical conductance and totally prevented the IFN-gamma-induced increase in mannitol and inulin fluxes. IL-10 did not prevent the IFN-gamma-induced abolishment of tight junctional charge selectivity but did attenuate the total increase in sodium and chloride permeability. IFN-gamma and IL-10 both separately reduced peak forskolin and carbachol-stimulated Isc. IL-10 pretreatment further enhanced the IFN-gamma-induced reduction in secretagogue-induced Isc. CONCLUSIONS: In T84 epithelial monolayers, IL-10 maintains the size, but not the charge, selectivity of the epithelial tight junction in the presence of IFN-gamma. In addition, both IL-10 and IFN-gamma limit carbachol and forskolin-induced increase in Isc.

Carbachol↗