Effects of succinylacetone on methyl alpha-D-glucoside uptake by the rat renal tubule.

Succinylacetone, a catabolic end-product of tyrosine, is excreted in large quantities in urine from individuals with hereditary tyrosinemia and the Fanconi syndrome. Succinylacetone inhibits rat renal tubular concentrative uptake of the glucose transport analogue, methyl alpha-D-glucoside, in a noncompetitive and reversible fashion. This compound also depresses oxygen consumption by the rat renal tubule without fine structural damage to mitochondria. It is concluded that succinylacetone may be a useful probe in elucidation of the biochemical mechanism underlying the human Fanconi syndrome.

Renal Fanconi syndrome: developmental basis for a new animal model with relevance to human disease.

Using succinylacetone (SA), a metabolite of tyrosine excreted in excess by infants and children with hereditary tyrosinemia and the renal Fanconi syndrome (FS), we have investigated developmentally-related membrane transport events leading to emergence of the generalized renal tubular dysfunction seen in human FS. SA was found to impair sugar and amino acid uptake by both newborn renal tubules and 7-day renal brush-border membrane vesicles (BBMV). This impairment by SA was due in part to a slowing of substrate cotransport rate of 22Na+-entry into BBMV. Concentration-dependent uptake studies indicated SA inhibited the newborn high-affinity transport systems for sugars and amino acids. SA also caused an increase in membrane fluidity and a shift in the thermotropic transition temperature. The demonstrated dual nature of SA's effect on membrane fluidity and O2 consumption, together with the relative contribution of each component to SA-induced transport impairment helps to provide a basis for an understanding of the age-related increases in glucosuria, aminoaciduria and natriuria seen in infants with FS.

Evaluation of a self-calibrating blood glucose monitor.

A new reflectance blood glucose meter designed for hospital use, the Reflocheck (Boehringer-Mannheim, West Germany), was evaluated. The instrument showed excellent correlation with a routine laboratory method (r = 0.996), and the coefficients of variation at high, medium, and low glucose levels were less than 4%.

Effects of succinylacetone on amino acid uptake in the rat kidney.

Infants with hereditary tyrosinemia also have a renal Fanconi syndrome and excrete succinylacetone (SA). We have studied the effects of SA on rat renal tubular amino acid transport in vivo and in vitro using isolated renal tubules. Injection of SA produces increased clearance of several amino acids in the intact animal. In vitro SA causes a reversible inhibition of alpha-aminoisobutyric acid uptake, resulting from depressed low- and high-affinity transport systems. Addition of glutamate, succinate, or glucose, alone or in combination, did not restore transport. These observations suggest the usefulness of SA in the production of a physiologic animal model for the study of the human Fanconi syndrome.

On the uptake of biotin by the rat renal tubule.

Little is known of biotin handling by transporting epithelium. Accordingly, we have examined the characteristics of biotin uptake by rat renal tubular epithelium. Renal cortical slices showed concentrative, temperature-sensitive uptake of biotin. Renal brushborder membrane vesicles exhibited an "overshoot" phenomenon with uptake of 1.9 nM biotin in the presence of a 100 mM NaCl gradient. This overshoot was reduced in magnitude with reduction of the sodium gradient to 50 mM. Biocytin significantly reduced uptake by the vesicles. Concentration-dependent studies yielded an apparent transport Km of 200 nM. We conclude that biotin is actively transported by the rat renal proximal tubule by a system which is at least partially Na+ dependent, and shared by biocytin.

Filtration coefficient and osmotic reflection coefficient to albumin in rabbit submandibular gland capillaries.

1. The isolated perfused submandibular salivary gland of the rabbit has been used in order to make estimates of the filtration coefficient (Kf) and reflection coefficient (sigma d) of the capillary wall to albumin. 2. An isogravimetric preparation was used and in paired experiments the value for Kf obtained in glands perfused with albumin-Krebs solution, 0.96 +/- 0.086 (mean +/- S.E. of mean) ml min-1 mmHg-1 100 g-1, was not significantly different from that in blood-perfused glands, 0.90 +/- 0.15. 3. On analysing the data for reflection coefficient, it was concluded that the above values underestimated Kf by about 30%; using corrected values for Kf, osmotic reflection coefficients were determined from the weight changes following a sudden change in the oncotic pressure of the perfusate. The value for sigma d to albumin lay between 0.79 and 1.0, the lower value being obtained after the Kf correction. 4. The high hydraulic conductivity, combined with sieving properties comparable to those in continuous capillaries, is consistent with other data on fenestrated capillaries. 5. Finally, it was observed that, while the Kf value calculated from the initial flux rate was similar whether measured during fluid efflux from or influx into the microvasculature, on returning to the initial conditions after raising osmotic pressure, efflux was now more rapid than influx. This phenomenon is discussed in relation to readjustment of Starling forces and the possible existence of an asymmetric double membrane in the capillary, interstitium system and cells of the salivary gland.

delta-Aminolevulinic acid dehydratase: is there a form unique to renal cortex?

Succinylacetone (SA) is known to be a potent inhibitor of delta-aminolevulinic acid dehydratase (ALAD) in the liver. We have examined the effects of SA on the rat renal cortical enzyme, our observations indicating very different behaviour of renal versus hepatic ALAD with SA treatment. While the temperature response of ALAD in both tissues was similar, addition of 4 mmol/l SA inhibited liver ALAD at 37 and 55 degrees C and enhanced renal ALAD activity 2- to 3-fold at each temperature. This increase in renal ALAD was progressive with SA concentrations form 1 to 10 mmol/l. A pH titration curve for both liver and kidney ALAD showed the hepatic enzyme to have a single pH optimum, while the renal enzyme had two, each of which was distinct from that in liver. Kinetic studies with and without 4 mmol/l SA over a 50-fold ALA concentration range indicated SA-induced enhancement of renal ALAD over the entire range at both pH optima. Using 14C-labelled ALA, we have confirmed these observations made on the basis of a colorimetric assay for PBG, the enzyme product. We conclude that renal ALAD may be a different molecular species from the liver enzyme. Further studies may clarify the significance of these observations to renal heme synthesis.

On rat renal aminolevulinate transport and metabolism in experimental Fanconi syndrome.

Hereditary tyrosinemia, an autosomal recessive disease of human infants, is characterized by severe liver disease, a renal Fanconi syndrome, and urinary excretion of large quantities of both aminolevulinate (ALA) and succinylacetone (SA). The latter is a metabolic end-product of tyrosine catabolism in affected individuals, produced by both liver and kidney, and is a potent inhibitor of aminolevulinate dehydratase (ALAD) in liver. This inhibition has been assumed to result in release of large amounts of aminolevulinate from liver into the circulation, with subsequent urinary excretion. In the present report we examine the effects of succinylacetone on rat renal cortical tubular handling of ALA and the relationship to tubular heme content, demonstrating a marked impairment of each. In contrast, maleic acid was found to have no effect on either renal ALAD or heme content. Thus, we conclude that renal handling of ALA in SA-treated rat renal cortex may indicate a contribution by the kidney to the increased net ALA excretion observed in hereditary tyrosinemia.

Effects of succinylacetone on the uptake of sugars and amino acids by brush border vesicles.

Infants with hereditary tyrosinemia excrete succinylacetone (SA) in their urine, and suffer from a reversible renal Fanconi syndrome with glycosuria and hyperaminoaciduria. Thus, we have examined the effects of 4 mM SA on rat renal brush border membrane vesicle uptake of sugars and amino acids. SA, unlike sodium maleate, significantly inhibits Na+-dependent vesicular sugar and amino acid uptake. 22Na-uptake, as well as membrane fluidity of the vesicles, are also affected by SA. Inhibition of glycine uptake by SA is reversible and competitive in nature, while alpha-CH3-D-glucoside uptake is non-competitively affected. We conclude, therefore, that SA has a more complex action on the rat renal tubule than sodium maleate, and is likely a much more physiologic model for study of the human renal Fanconi syndrome.

Impaired microvascular hyperaemic response to minor skin trauma in type I diabetes.

The microvascular response of foot skin to minor thermal injury and the skin of the anterior abdominal wall to injury from a needle was assessed by laser Doppler flowmetry in 23 patients with type I diabetes and 21 healthy control subjects. After minor thermal injury mean (SD) maximum skin blood flow was significantly lower in the diabetic group than the control group (0.53 (0.11) v 0.72 (0.10) V, in arbitrary units of flow, respectively, p less than 0.001) and was negatively correlated with the duration of diabetes (r = -0.60; p less than 0.01). After needle injury a similar pattern of impairment was seen, the peak flow value recorded being significantly lower in the diabetic group than the control group (0.28 (0.10) v 0.41 (0.09) V, respectively; p less than 0.001) and also negatively correlated with the duration of diabetes (r = -0.61; p less than 0.01). There was a significant relation between the response obtained at the two sites of injury in the diabetic group (r = +0.72, p less than 0.001) but not in the control group. The impairment in response was not related to diabetic control and was not explicable in terms of a reduction in superficial skin capillary density. The inability of the diabetic skin microvasculature to respond normally to injury may be an important factor in the development of foot ulceration that often follows minor trauma.

Direct observation of orbital ordering in La0.5Sr1.5MnO4 using soft x-ray diffraction.

We report the first direct resonant soft x-ray scattering observations of orbital ordering. We have studied the low temperature phase of La0.5Sr1.5MnO4, a compound that displays charge and orbital ordering. Previous claims of orbital ordering in such materials have relied on observations at the manganese K edge. These claims have been questioned in several theoretical studies. Instead we have employed resonant soft x-ray scattering at the manganese L(III) and L(II) edges which probes the orbital ordering directly. Energy scans at constant wave vector are compared to theoretical predictions and suggest that at all temperatures there are two separate contributions to the scattering: direct orbital ordering and strong cooperative Jahn-Teller distortions of the Mn3+ ions.