Biomedical subjects

T D Dubose

Publications and source records attributed to T D Dubose.

6 recordsLinked to original sources

PubMed2000

Sch-28080 depletes intracellular ATP selectively in mIMCD-3 cells.

Two H(+)-K(+)-ATPase isoforms are present in kidney: the gastric, highly sensitive to Sch-28080, and the colonic, partially sensitive to ouabain. Upregulation of Sch-28080-sensitive H(+)-K(+)-ATPase, or "gastric" H(+)-K(+)-ATPase, has been demonstrated in hypokalemic rat inner medullary collecting duct cells (IMCDs). Nevertheless, only colonic H(+)-K(+)-ATPase mRNA and protein abundance increase in this condition. This study was designed to determine whether Sch-28080 inhibits transporters other than the gastric H(+)-K(+)-ATPase. In the presence of bumetanide, Sch-28080 (200 microM) and ouabain (2 mM) inhibited (86)Rb(+) uptake (>90%). That (86)Rb(+) uptake was almost completely abolished by Sch-28080 indicates an effect of this agent on the Na(+)-K(+)-ATPase. ATPase assays in membranes, or lysed cells, demonstrated sensitivity to ouabain but not Sch-28080. Thus the inhibitory effect of Sch-28080 was dependent on cell integrity. (86)Rb(+)-uptake studies without bumetanide demonstrated that ouabain inhibited activity by only 50%. Addition of Sch-28080 (200 microM) blocked all residual activity. Intracellular ATP declined after Sch-28080 (200 microM) but recovered after removal of this agent. In conclusion, high concentrations of Sch-28080 inhibit K(+)-ATPase activity in mouse IMCD-3 (mIMCD-3) cells as a result of ATP depletion.

Adenosine Triphosphate↗

PubMed1999

Peptides derived from the human transferrin receptor stimulate endosomal acidification via a Gi-type protein.

UNLABELLED: Peptides derived from the human transferrin receptor stimulate endosomal acidification via a Gi-type protein. BACKGROUND: Acidification of the endosomal compartment is a prerequisite for intracellular processing of endocytosed complexes. Endosomal acidification is accomplished by an H+-ATPase, in parallel with a Cl- conductance. Previous studies from our laboratory have demonstrated that endosomal acidification is modulated by a pertussis toxin-sensitive mechanism, suggesting that endosomal acidification could be regulated through a self-contained signal transduction pathway. This study was designed to test this hypothesis using the transferrin receptor as a model. METHODS: Synthetic peptides corresponding to a region of the cytosolic domain of the transferrin receptor and containing a KPKR sequence were used to stimulate endosomal acidification in a G-protein-dependent manner. RESULTS: Peptides activated the Gi, as evidenced by stimulation of the rate of GTPgammaS binding. A transferrin receptor peptide that lacked the KPKR sequence did not stimulate endosomal acidification and failed to promote GTPgammaS binding to Gi proteins. CONCLUSIONS: These results demonstrate that regulation of endosomal acidification can be achieved, in part, through a Gi-mediated signal transduction pathway. These findings suggest that regulation of endosomal acidification through such a pathway may facilitate intracellular processing of the transferrin receptor.

Amino Acid Sequence↗

PubMed1997

Mannitol-induced acute renal failure.

The osmotic diuretic mannitol may be used in diverse clinical settings, such as providing "renal protection" in patients at risk for acute renal failure, decreasing intracranial pressure in patients with intracranial trauma, and preventing the dialysis-disequilibrium syndrome. Mannitol is commonly used after cardiac catheterization, cardiovascular surgery, and exposure to intravenous contrast dyes. This study presents a case in which a long-term renal transplant recipient receiving cyclosporine therapy concomitantly developed acute renal failure after the administration of high-dose mannitol in an attempt to induce an osmotic diuresis. The diagnosis of "osmotic nephrosis" was confirmed by renal biopsy, and the condition was reversed by cessation of the agent. Studies in experimental animals indicate that cyclosporin A can potentiate the tubular toxicity of mannitol, but such an association has not been verified in humans. Numerous studies confirm the nephrotoxic potential of high-dose mannitol, especially in patients with renal insufficiency. The clinical utility of the osmolar gap in preventing mannitol nephrotoxicity is emphasized.

Acute Kidney Injury↗

PubMed1993

Hyponatremia in the cancer patient.

Hyponatremia is a common metabolic complication in cancer patients. It can be caused by the primary tumor itself, metastasis, by diagnostic or therapeutic interventions, or by a secondary complication. A clear understanding of water and sodium homeostasis is required for evaluation. The initial diagnostic step is to define a hypoosmolar state. The history and physical examination should be tailored to determine the volume status of the patient--volume depleted, euvolemic, or edematous. In general, hyponatremia associated with volume depletion is treated with saline, while fluid restriction is the primary therapy in patients with normal or increased extracellular fluid volume. In symptomatic patients, the duration of the hyponatremia and its causes should be determined. Both overzealous and inadequate treatment put the patient at risk of serious neurologic sequelae.

Humans↗

PubMed1989

Cytoplasmic pH in pulmonary macrophages: recovery from acid load is Na+ independent and NEM sensitive.

The pulmonary macrophage plays a primary role in the immunological defense of the lung. Although many studies have been devoted to elucidation of its phagocytic and secretory functions, little is known of its membrane transport properties or of how it regulates intracellular pH (pHi). The purpose of this study, therefore, was to determine base-line pHi and the mechanism(s) by which the cell recovers pHi when challenged with an intracellular acid load. Through the use of the pH-sensitive fluorescent dye, 2,7-biscarboxyethyl-5(6)-carboxy-fluorescein (BCECF), base-line pHi was estimated to be 7.24 +/- 0.03. Cells were acidified by two methods, nigericin and weak acids, while recovery (dpHi/dt) was monitored. The rate of recovery was found to be independent of external Na+ and K+ and was insensitive to amiloride. Pretreatment with 4,4'-diiso-thiocyanatostilbene-2,2'-disulfonic acid, an inhibitor of Cl- -HCO3- exchange, was also without effect on recovery from an intracellular acid load in these cells, under nominally HCO3- -free conditions. In contrast, N-ethylmaleimide (NEM) and N,N'-dicyclohexylcarbodiimide, nonspecific inhibitors of proton adenosinetriphosphatases (ATPases), virtually abolished pHi recovery. Efflux of H+ equivalents by pulmonary macrophages was measured by techniques involving both pH stat titration and the effect on fluorescence of extracellular BCECF. Basal H+ extrusion was approximately 2.75 +/- 0.64 nmol H+.min-1.10(6) cells-1 and was enhanced to approximately 26.0 +/- 6.95 nmol H+.min-1.10(6) cells-1 in acid-loaded cell suspensions. The basal rate of H+ extrusion was reduced to approximately 0.84 +/- 0.31 nmol H+.min-1.10(6) cells-1 in the presence of 1 mM NEM. These results suggest that recovery of cytoplasmic pH from an intracellular acid load, as well as regulation of pHi, under the conditions examined, is not mediated by a Na+-H+ exchanger in these cells. Rather, the data are consistent with the presence of an H+-ATPase in the plasma membrane of pulmonary macrophages.

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

PubMed1983

Effects of acetate and bicarbonate hemodialysis on cardiac function in chronic dialysis patients.

Previous studies have suggested that acetate hemodialysis causes myocardial depression. This study examines the acute effects of hemodialysis using, alternately, bicarbonate and acetate in the dialysate, on cardiac function in ten patients. These patients were also studied during acetate dialysis using a large surface area (SA) dialyzer. Each patient was dialyzed for 4 hr with: (1) 1.0 m2 SA dialyzer and bicarbonate dialysate; (2) 1.0 m2 SA dialyzer and acetate bath, and; (3) 2.5 m2 SA dialyzer and acetate dialysate. All studies were performed during isovolemic dialysis to separate the effects of changes in cardiac filling volume with hemodialysis, from changes in myocardial contractility. Myocardial function, as assessed by pre- and postdialysis echocardiographically derived fractional shortening (Fs) and mean velocity of circumferential shortening (VCF), improved (P less than 0.05) to the same extent, after all three dialysis treatments. This occurred despite greater increases (P less than 0.002) in pH and bicarbonate after bicarbonate dialysis and decreases (P less than 0.05) in PO2, PCO2 and bicarbonate after acetate dialysis with 2.5 m2 SA dialyzer. These results indicate that diffusive dialysis with both acetate and bicarbonate dialysate improves myocardial function and do not support the view that acetate influx during dialysis can lead to myocardial depression.

Acetates↗