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Biomedical subjects

E Kenyon

Publications and source records attributed to E Kenyon.

14 recordsLinked to original sources
PubMed1997

Apical and basolateral membrane mechanisms that regulate pHi in bovine retinal pigment epithelium.

pH regulation was studied in fresh explant bovine retinal pigment epithelium-choroid using the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and intracellular microelectrodes. Acid recovery was HCO3 dependent, inhibited by apical amiloride and apical or basal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), and required apical and basal Na. Alkali recovery was HCO3 dependent and inhibitable by apical or basal DIDS. Three apical and two basolateral transporters were identified. Four contribute to acid extrusion, i.e., apical Na/H exchange, apical H-lactate cotransport, and apical Na-HCO3 cotransport and basolateral Na-HCO3 cotransport. At least two contribute to alkali extrusion, i.e., apical Na-HCO3 cotransport and a basolateral HCO3-dependent, DIDS-inhibitable mechanism, possibly Na-HCO3 cotransport, Cl/HCO3 exchange, or both. The apical Na-HCO3 cotransporter is electrogenic, carrying net negative charge inward. Basal Cl removal or addition of basal HCO3 caused HCO3- and Cl-dependent alkalinizations, respectively. Apical DIDS increased both responses. These cytosolic pH (pHi) regulatory mechanisms are so tightly coupled that changes in pHi can only occur after two or more of them are inhibited. In addition, these mechanisms help provide pathways for transport of Na and HCO3 across the retinal pigment epithelium between the blood and the distal retina.

Acids↗
PubMed1994

Lactate transport mechanisms at apical and basolateral membranes of bovine retinal pigment epithelium.

The isolated bovine retinal pigment epithelium actively transports lactate from the apical to the basal bath. Net short-circuit [14C]lactate flux in 20 mM lactate was 0.46 +/- 0.09 mu eq.cm-2.h-1 (n = 8). In open circuit, with a physiological lactate gradient, net [14C]lactate flux was 0.66-1.31 mu eq.cm-2.h-1 (n = 3). Lactate in the apical bath caused intracellular acidifications that were saturable, apparently stereospecific, and reduced in magnitude by several H-lactate cotransport inhibitors. In the basal bath, lactate caused intracellular alkalinizations that were dependent on the presence of Na. In short circuit, 20 mM lactate in both baths reversed the direction of net transepithelial 22Na transport from secretion to absorption, suggesting the presence of basolateral Na-lactate cotransport moving lactate out of the cells. Outwardly directed Na-lactate cotransport requires a lactate:Na stoichiometry > 1.4:1, consistent with the coupled movement of Na, lactate, and net negative charge across the basolateral membrane. Intracellular microelectrode recordings showed that basal lactate hyperpolarized and apical lactate depolarized the basolateral membrane. For lactate absorption, this is a novel arrangement of membrane proteins:luminal H-lactate cotransport and serosal electrogenic Na:(n)lactate cotransport. Lactate transport across the retinal pigment epithelium may play an important role in regulating retinal metabolism and subretinal space volume and composition.

Animals↗
PubMed1994

Lactate transport in freshly isolated human fetal retinal pigment epithelium.

PURPOSE: To study transport mechanisms for small monocarboxylic acids in the apical and basolateral membranes of freshly isolated, human fetal retinal pigment epithelium. METHODS: The epithelium was mounted in a small Ussing chamber that allowed separate perfusion of both the apical and basal compartments and simultaneous measurements of intracellular pH, transepithelial potential, and tissue resistance. Intracellular pH was measured using a pH-sensitive dye, 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein. RESULTS: When 10-100 mM lactate or pyruvate was added to the apical bath the cells acidified by 0.10-0.25 pH units. There were no differences between the initial rates of intracellular acidification produced by L-lactate and D-lactate. These rates could be described as Michaelis-Menten functions of the concentrations of lactate and pyruvate. The Km values were 42 +/- 12 mM for L-lactate and 34 +/- 8 mM for pyruvate. The rates of acidification caused by 50 mM L-lactate were reversibly reduced by 44% or 35% after apical administration of probenecid (2 mM) or alpha-cyano-4-hydroxycinnamate (2 mM), and irreversibly reduced by 78% after apical administration of the sulfhydryl-reagent mersalyl acid (2 mM). The intracellular acidifications caused by apical pyruvate (50 mM) were completely and reversibly inhibited by 50 mM apical L-lactate. Addition of 50 to 100 mM lactate to the basal bath caused intracellular alkalinizations, which could be inhibited by Na+ removal in the basal bath or by 2 mM alpha-cyano-4-hydroxycinnamate in the apical bath.

Basement Membrane↗
PubMed1992

Na-dependent pHi regulatory mechanisms in native human retinal pigment epithelium.

This study provides the first information about pHi regulatory mechanisms in human retinal pigment epithelium (RPE). The experiments were carried out on fresh explant tissues from adult donor and fetal eyes, and pHi was measured using fluorescence microscopy techniques and the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. In adult donor RPE, the resting pHi is 7.30 +/- 0.14 (mean +/- standard deviation; n = 6) in HCO3 Ringer's solution. In HCO3 Ringer's solution, apical Na removal caused rapid cell acidification with an initial rate of 0.40 +/- 0.10 pH U/min (n = 4). This Na-dependent acidification was partially inhibited by apical amiloride (n = 1) and DIDS (n = 1). In HCO3 Ringer's solution, pHi recovery from an acid load (NH4 prepulse) also was blocked by apical Na removal. In nominally HCO3-free Ringer's solution, apical amiloride (1 mmol/l) acidified the cells. These results suggest that the apical membrane of adult human RPE contains an Na/H exchanger and possibly a Na-dependent, DIDS-inhibitable pH regulatory mechanism, perhaps a NaHCO3 cotransporter. For the fetal RPE, the resting pHi was 7.16 +/- 0.10 (n = 9) and 7.19 +/- 0.10 (n = 20) in HCO3 and HCO3-free Ringer's solution, respectively. In HCO3 and HCO3-free Ringer's solution, apical amiloride (1 mmol/l) acidified the cells and the removal of apical Na caused cell acidification with an initial rate of 0.30 +/- 0.08 (n = 32) and 0.58 +/- 0.29 (n = 6) pH U/min, respectively. The pHi recovery from an acid load also was blocked by apical amiloride and apical Na removal. These results suggest that the apical membrane Na/H exchanger is the dominant acid extrusion mechanism in human fetal RPE.(ABSTRACT TRUNCATED AT 250 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1988

Corneal deswelling following overnight wear of rigid and hydrogel contact lenses.

The edema response over a 24-hour sleep/wake cycle of ten subjects wearing a rigid gas-permeable (RGP) lens in one eye and a hydrogel lens in the other was evaluated. Lenses that result in equivalent amounts of overnight edema were selected. There was no significant difference in the rates of deswelling during the initial hour. However, at three and five hours after eye opening, the amount of residual edema was greater in the eye wearing the hydrogel lens. We suggest that when an RGP lens provides the same closed-eye level of oxygen as a hydrogel lens, the greater tear exchange of the RGP lens will result in a greater open-eye oxygen supply, leading to less daytime edema. This suggests that the more complete deswelling we observed with RGP lenses can be attributed to a lower stimulus to open-eye swelling. Biomicroscopy, subjective vision, and patient comfort were also rated on waking and ten hours after eye opening. On eye opening there was significantly more back-surface debris (P = 0.01) with the RGP lenses. Patients wearing RGPs rated comfort as poorer and vision as better but only the latter attained statistical significance.

Adolescent↗
PubMed1988

Rigid contact lens adherence: incidence, severity and recovery.

The incidence of rigid lens adherence in one eye of 25 subjects was compared to the results for the fellow eye wearing a soft lens of nearly equal permeability in an in-laboratory overnight study. A 48% incidence of adherence was found for both extended wear modalities, which demonstrates the high frequency of this problem. Adherence of both rigid and soft lenses caused epithelial breakdown and corneal distortion, but these sequelae were generally more severe with rigid lenses. Recovery data on nine subjects whose rigid lenses adhered show that positive signs of adherence often disappear within 1 hour, demonstrating the need for early morning examinations. Practical considerations for the detection of rigid lens adherence and the possible consequences of failure to diagnose this condition are discussed.

Adhesiveness↗
PubMed1988

RGP lens binding.

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Contact Lenses, Extended-Wear↗
PubMed1987

The occurrence of chemically induced hormesis.

This paper will provide an overview of the potential general applicability of chemical hormesis. Evidence will be presented on the occurrence of chemical hormesis by chemical classes studied (e.g. heavy metals, polycylic aromatic hydrocarbons, etc.), by affected biological and toxic end points (e.g. growth enzyme activities, DNA-repair capacity, life span, tumor incidence) and by biological/taxonomic systems. Several specific examples of possible hormetic effects with respect to agents of environmental concern, such as chloroform, will be presented along with a discussion of future research directions.

Animals↗
PubMed1986

Influence of wearing schedule on extended-wear complications.

The effect of wearing schedule on extended-wear (EW) success was evaluated over a nine-month period by assigning 36 subjects to 5 different wearing schedules (removal every 4, 7, 14, or 28 days, or daily wear [DW]). All but one EW patient required interruption of extended wear at least once; half of the DW patients completed the study without interruption. The most prevalent (100%) complication in the EW group was epithelial microcysts; 54% had to discontinue EW for 67 +/- 30 days while severe microcysts subsided. Evidence is given for a hypoxic etiology of microcysts. Other EW complications included: persistent punctate keratitis, infiltrative keratitis, infection, and idiopathic red eye. Neither the level of overnight corneal swelling nor the period between removals influenced the incidence or severity of complications. Data is given on the recovery of corneal parameters after EW is discontinued.

Adult↗
PubMed1977

The behaviour of thyroid hormones in an infant with untreated neonatal thyrotoxicosis.

The clinical course from birth and serial measurements of serum T3, T4 and TSH in an infant with untreated neonatal thyrotoxicosis are reported. The thyroid hormone levels fell exponentially with time at rates very much slower than those previously reported for the maternally-transmitted thyroid stimulating antibody generally thought to cause the disorder. Steady physiological levels of thyroid hormones were achieved after 110 days (serum T3 = 3.4 NMOL/L, T4 = 118 nmol/l). TSH first rose to a measurable level after about 90 days.

Adult↗