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Biomedical subjects

R Green

Publications and source records attributed to R Green.

At least 307 records · Page 17Linked to original sources
PubMed1988

Conservation of central nervous system glutaryl-coenzyme A dehydrogenase in fruit-eating bats with glutaric aciduria and deficient hepatic glutaryl-coenzyme A dehydrogenase.

The adult fruit-eating bat, Rousettus aegypticus, excretes massive amounts of glutaric acid in the urine (20-70 mumol/mg creatinine) comparable to those of humans affected with the inherited metabolic disorder, glutaric aciduria type I. Glutaric acid was quantified by sequential liquid partition chromatography and gas chromatography. Oral loading with the amino acid precursors of glutaric acid, L-lysine and L-tryptophan, resulted in significant increases in glutaric acid excretion above the base-line values. Glutaryl-CoA dehydrogenase activity was assayed in adult bat tissues and compared with the same tissues in the rat using methods of 14CO2 evolution from 1,5-[14C]glutaryl-CoA. A severe deficiency of glutaryl-CoA dehydrogenase activity was found in the bat liver and kidney, whereas brain and spinal cord levels were similar to those in the rat. Reverse phase high performance liquid chromatography analysis of the metabolites in the assay mixture showed negligible hydrolysis of [14C]glutaryl-CoA to free [14C]glutaric acid and complete conversion of the product [14C]crotonyl-CoA to 3-hydroxy[14C]butyryl-CoA. The adult bat, with its huge glutaric acid excretion and deficient liver glutaryl-CoA dehydrogenase, metabolically mimics patients affected with glutaric aciduria type I. The bat does not, however, display the neurologic manifestations seen in patients. This may be explained by conservation of glutaryl-CoA dehydrogenase activity in the central nervous system of the bat.

Animals↗
PubMed1988

Biotin-dependent expression of the asialoglycoprotein receptor in HepG2.

The asialoglycoprotein receptor (AsGR) is characteristic of fully differentiated hepatocytes. AsGR expression in confluent cultures of HepG2 cells grown in minimal essential medium (MEM) requires a 300-350-dalton dialyzable fraction of fetal bovine serum (FBS). Addition to dialyzed FBS (dFBS) of 10(-7) M biotin or biocytin (Mr 372) permitted full expression of AsGR by HepG2. Affinity chromatography of FBS on streptavidin-Sepharose abolished its ability to support AsGR production. The bound material, when released by heat denaturation and resolved by thin layer chromatography, yielded three cinnamaldehyde-positive components, of which the major detectable one migrates with authentic biocytin and reconstitutes dFBS. Sera from several species, which do not support AsGR production by HepG2, contain less than 10% biotin found in FBS as determined by direct enzyme-linked immunosorbent assay. These results indicate that biotin or a derivative is the low molecular weight serum factor of FBS required for expression of AsGR. Isolation of messenger RNA from HepG2 revealed no difference in AsGR transcripts when cells were grown in MEM-10% FBS or MEM-10% dFBS. Thus a biotin-dependent post-transcriptional event permits the ultimate expression of the AsGR by HepG2 cells.

Actins↗
PubMed1988

Effect of a meal containing protein on lithium clearance and plasma immunoreactive atrial natriuretic peptide in man.

1. The effect of meals with a high and low protein content and of the fasting state on renal function and plasma atrial natriuretic peptide was studied in water-loaded normal volunteers. 2. Creatinine clearance increased after the high protein meal, but did not change after the low protein meal or while fasting. Observations of similar increases in urine sodium and potassium excretion and a transient decrease in urine flow after both meals suggest that the protein content of the meal is not an important contributory factor in these responses to feeding. 3. Absolute delivery of sodium and water out of the proximal tubules (assessed by the lithium clearance method) was higher after both meals than while fasting; fractional lithium clearance was higher after the low protein meal than the high protein meal and while fasting. Absolute reabsorption from proximal tubules was increased after only the high protein meal. 4. A transient decrease in the fraction of water delivered to distal nephron segments that appeared in the urine (fractional distal water excretion) was observed after both meals. Fractional distal sodium excretion and absolute distal sodium and water reabsorption increased after both meals. 5. Since plasma atrial natriuretic peptide either decreased (high protein meal) or remained unchanged (low protein meal and fasting), it is unlikely that this hormone is involved in the hyperfiltration after the high protein meal and the natriuresis after both high and low protein meals.

Adult↗
PubMed1988

Effect of low dose infusion of atrial natriuretic peptide on renal function in man.

1. The effects of the infusion of a low dose (2 pmol min-1 kg-1 for 3 h) of human atrial natriuretic peptide (hANP) were studied in seven healthy volunteers undergoing a water diuresis. Lithium clearance was used to monitor proximal tubular function. 2. hANP increased urine flow rate, sodium, calcium and magnesium excretion without significant changes in potassium and phosphate excretion, heart rate or blood pressure. 3. hANP caused a small change in fractional lithium clearance, and larger changes in distal nephron handling of sodium and water. 4. Plasma renin activity tended to decrease during the infusion of hANP, while plasma aldosterone concentration decreased during and increased after stopping the infusion of hANP. 5. The data suggest that hANP inhibits the reabsorption of sodium and water by an action on distal segments of the nephron and perhaps the proximal tubule. Inhibition of renin and aldosterone secretion may contribute to the natriuresis.

Adult↗
PubMed1988

The effect of DIDS on fluid reabsorption from the proximal convoluted tubule of the rat: dependence on the presence of bicarbonate.

The effects of the anion transport inhibitor DIDS (4,4'-diisothiocyano-stilbene-2,2'-disulphonate) on fluid reabsorption by the rat proximal convoluted tubule were investigated by the stationary microperfusion (split-drop) method. Addition of 10(-5) mol 1-1 DIDS to luminal and capillary perfusates in the presence of peritubular bicarbonate (25 mmol 1-1) increased the reabsorptive half-time (t 1/2) from 35.1 +/- 4.5 to 57.0 +/- 6.2 s, corresponding to a decrease in fluid reabsorption (Jv) from 1.33 +/- 0.06 to 0.89 +/- 0.09 nl min-1 mm-1. Omission of bicarbonate per se from the peritubular capillaries reduced Jv to 0.41 +/- 0.03 nl min-1 mm-1 and this reabsorptive flux was completely inhibited by the addition of 2 mmol 1-1 NaCN to both the luminal and peritubular perfusates. In the absence of bicarbonate, the addition of DIDS at either 10(-4) or 10(-5) mol 1-1 to both lumen and capillary perfusates had no significant effect on either t 1/2 or Jv. These results show that the inhibitory action of DIDS on fluid reabsorption is dependent on the presence of bicarbonate. However, the proximal tubule also reabsorbs water in the absence of bicarbonate by a mechanism that is dependent on cellular metabolism, but is insensitive to DIDS at the concentrations used in this study.

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

The effects of anions on fluid reabsorption from the proximal convoluted tubule of the rat.

1. Fluid reabsorption from surface proximal tubules of the rat was measured in vivo using stationary microperfusion techniques. Reabsorptive rate (Jv) was measured from droplets containing chloride as the main reabsorbable anion and when chloride was substituted by bromide, iodide, nitrate, acetate, isethionate or methylsulphate in either the tubular lumen alone or in both lumen and peritubular capillaries. 2. In tubules with an intact blood supply, droplet volume decreased in a manner best described by a single exponential and substitution of chloride by nitrate or bromide had no effect on Jv. Substitution by iodide or acetate inhibited Jv by approximately 17% but substitution by methylsulphate or isethionate caused droplets to transiently increase in volume before shrinkage which was itself inhibited by approximately 50%. The inhibitory action of isethionate was found to be concentration dependent. 3. Recollection and analysis of droplets which were initially free of chloride, containing either nitrate or isethionate, showed that chloride entered these droplets, but that the initial rate of chloride entry was greater for nitrate than isethionate droplets. 4. When tubules and capillaries were perfused with chloride solutions containing no bicarbonate, Jv was reduced to about 20% of the value when peritubular capillary blood flow was intact. Substituting chloride in the tubular and capillary perfusion revealed a sequence for supporting fluid reabsorption that was identical to that when chloride was substituted in tubule fluid alone: bromide = nitrate greater than iodide = acetate greater than isethionate. Addition of 2.0 mmol l-1 NaCN reduced the reabsorptive flux to zero. 5. The results of this study are consistent with transcellular transport of anions across the proximal tubular epithelium. The pathways for anion transport are likely to involve a series of non-selective mechanisms such as anion exchangers.

Absorption↗
PubMed1988

Chloride transport in the proximal renal tubule.

Our knowledge of chloride transport along the nephron has greatly expanded. Whereas for a long time it was assumed that chloride ions were reabsorbed entirely passively with sodium--the "mendicant" role of chloride, more recent studies suggest that several distinct reabsorptive transport mechanisms operate in parallel. Accordingly, a new model of proximal chloride transport has evolved that includes both active, transcellular as well as passive, intercellular transport pathways. Transcellular chloride reabsorption involves anion exchange mechanisms in both the luminal and peritubular cell membranes, processes that also depend on sodium, hydrogen, and bicarbonate ions. Chloride transport is thus intimately related to sodium and fluid transport as well as to cell acid-base metabolism. Unresolved problems concern the relative magnitude of transcellular and paracellular chloride transport and the details of luminal and basolateral chloride translocation steps.

Animals↗
PubMed1988

The relative accuracy of coupler and behavioural estimates of the real ear gain of a hearing aid.

Selection of a hearing aid frequency response is often initially based on coupler-measured performance, but is subsequently confirmed by functional measurement of real ear gain. It is usually assumed that functional gain measurement is necessary as confirmation because coupler measures do not take individual differences into account. This paper first argues from a theoretical standpoint that, if appropriate coupler/real ear correction factors are used, coupler estimates of real ear gain can be of comparable accuracy to functional estimates. It then uses experimental evidence from a study carried out by the author to show that, because of the variability in functional measurements of gain, suitably corrected coupler estimates of real ear gain can be more accurate than their functional counterparts.

Auditory Threshold↗
PubMed1987

Subcellular concentrations of estrone, estradiol, androstenedione and 17 beta-hydroxysteroid dehydrogenase (17-beta-OH-SDH) activity in malignant and non-malignant human breast tissues.

Total and subcellular (cytosol and nuclear) concentrations of estrone (E1), estradiol (E2), and androstenedione were determined in non-malignant (n = 61) and malignant (n = 65) human breast tissues obtained from post-menopausal women. The 17 beta-hydroxysteroid dehydrogenase (17 beta-OH-SDH) activity was determined in 800g supernatant fraction. Total estrogens, E1 and E2 levels and 17 beta-OH-SDH activity were significantly (p less than 0.005, 0.0005, 0.001, respectively) higher in malignant than in non-malignant breast tissues. We failed to observe significant changes in subcellular steroid concentrations or enzyme activity associated with patients' obesity or tumor estrogen receptor status. When the steroid levels were analyzed in relation to clinical staging of the disease, nuclear contents of estradiol were significantly higher (p less than 0.005) in Stage-IV patients than in those with less advanced disease (Stages I to III). 17 beta-OH-SDH activity was significantly (p less than 0.001) lower in patients with advanced disease than in those with relatively less advanced (Stages I to III) disease and was positively correlated with tissue concentration of androstenedione. Our present data indicate that differential intracellular metabolism of steroid hormones may have some influence on availability of estradiol at nuclear sites. In postmenopausal women, local interconversion of estrogens may provide sufficient estrogenic stimulus to enhance the growth and progression of breast tumors.

17-Hydroxysteroid Dehydrogenases↗
PubMed1987

Receptors for transferrin and transcobalamin II display segregated distribution on microvilli of leukemia L1210 cells.

Simultaneous addition of uniform latex particles derivatized with transferrin (0.532 micron) and transcobalamin II (0.345 micron) to leukemia L1210 cells resulted in segregated binding to individual microvilli as demonstrated by scanning electron microscopy. This segregated distribution suggests that individual microvilli are endowed either transferrin or transcobalamin II receptors but not both. Intracellular sorting and segregation of newly synthesized or recycling receptors probably occur prior to expression on the plasmalemma microvilli.

Animals↗
PubMed1987

A fluorescent dye which recognizes mature peripheral erythrocytes of myeloproliferative disorders.

Based on the previous finding that erythrocytes from patients with chronic myelogenous leukemia stain with the fluorescent dye merocyanine 540, erythrocytes from patients with other myeloproliferative disorders were examined for their ability to bind the membrane probe. As assessed by both fluorescence staining and a quantitative dye removal assay, all samples of erythrocytes from patients with chronic myelogenous leukemia, polycythemia vera, myelofibrosis with myeloid metaplasia and essential thrombocythemia bound more dye than did erythrocytes from normal, healthy individuals. Erythrocytes from three of six patients with acute myelogenous leukemia also showed increased affinity for the dye. In contrast, erythrocytes from three patients with acute lymphocytic leukemia and one with unclassifiable leukemia bound only normal amounts of dye. The procedures described may be useful as a supplemental aid to diagnosis of myeloproliferative disorders or for investigation of hematological diseases where multilineage involvement is suspected.

Cell Differentiation↗