A cobalamin metabolic defect with homocystinuria, methylmalonic aciduria and macrocytic anemia.

We have identified a patient with methylmalonic aciduria and homocystinuria due to a defect in cobalamin metabolism of the cb1C type mutant. At the time of admission at eight months of age the patient was malnourished, hypotonic and had macrocytic anemia. Neonatal screening for hypermethioninemia associated with homocystinuria had been normal. Serum vitamin B12 was markedly increased and folate concentration was above normal, as were urinary homocystine and methylmalonic acid. The patient had abnormal brain stem auditory and visual evoked potentials. Fibroblast activity of N5-methyltetrahydrofolate: homocysteine methyltransferase was reduced to approximately 10% of concurrent controls. A course of therapy with hydroxocobalamin resulted in a 90% reduction in excretion of methylmalonic acid and normalization of the evoked potentials. These studies support the efficacy of hydroxocobalamin therapy in this disease, suggest that methylmalonic acid may be the most appropriate metabolite to monitor for therapeutic response, and in importance of electrophysiologic studies in character in objectively monitoring the response to treatment metabolic disease.

Taurine uptake by cultured human lymphoblastoid cells.

Cultured human lymphoblastoid cells take up taurine from the medium by two processes: 1) a temperature-dependent, Na+-dependent, saturable "active"-transport system and 2) diffusion. The active transport has properties similar to those reported for taurine transport by other tissues. Apparent Km is about 25 microM and Vmax about 7.2 pmol/min/10(6) cells; saturation occurs at 100 microM taurine. Uptake is competitively inhibited by the beta-amino acids hypotaurine (50% inhibition at 44 microM) and beta-alanine (50% at 152 microM), as measured at 50 microM taurine. Taurocyamine inhibits 50% at 260 microM. Chlorpromazine and imipramine are strong uncompetitive inhibitors, giving 50% inhibition at 26 microM and 115 microM, respectively; at these concentrations cellular viability per se is not affected. Ouabain inhibits 40-50% over a concentration range of 4-500 microM. Diffusion of taurine into the cells is proportional to concentration up to 20 mM. However, at the concentration of taurine in human plasma, 40-100 microM, active transport would provide 90% of the taurine taken up.

Ornithine carbamoyltransferase deficiency in an adult male patient: significance of hepatic ultrastructure in clinical diagnosis.

Ornithine carbamoyltransferase (OCT) activity was deficient (8% of control) in the liver of a 21-year-old man who died after suddenly becoming comatose. Activities of other enzymes of the urea cycle in the liver were normal. There was no known prior illness or injury; the patient, however, had been taking liquid protein supplements to his diet. Hyperammonemia and orotic aciduria were present, and the concentration of lysine in the plasma was elevated. Survey of earlier reports indicates that neither the specific deficiency of hepatic OCT nor the urine and plasma findings provide a basis for definitive diagnosis of the patient's illness as primary OCT deficiency or as Reye's syndrome. Indeed, the age of the patient at onset of symptoms and the absence of any prodromal infection argue against the OCT deficiency being either primary or a sequel to Reye's syndrome. We suggest that it was secondary to mitochondrial injury caused by an unknown agent. Electron microscopic study of hepatocyte ultrastructure lends support to this view; abnormalities of the patient's mitochondria (bizarre, elongated shapes) do not resemble those seen in Reye's syndrome, nor have abnormalities been found in primary OCT deficiency.

Intermittent hypermethioninaemia associated with normal hepatic methionine adenosyltransferase activity: report of a case.

A 5-month-old infant was found to have hypermethioninaemia (0.8 mumol/ml) that has persisted intermittently (0.02-1.3 mumol/ml) over a period of 3 years. She presented with developmental delay and failure to thrive associated with gross abuse and neglect. Histological examination of the liver revealed inflammation of the portal triads. The activity of hepatic L-methionine-S-adenosyltransferase (EC 2.5.1.6) was normal. Whether the biochemical findings were the cause or the result of the hepatic damage is uncertain, but the minimal histological findings in the liver suggest a primary biochemical defect.

Methioninemia and myopathy: a new disorder.

A 7 1/2-year-old girl with hypermethioninemia, myopathy, and mental deficiency (IQ = 65) is described. The increased methionine was not associated with deficiency of methionine adenosyltransferase, which was normal or increased in liver, muscle, erythrocytes, and cultured fibroblasts. Methionyl-tRNA synthetase in fibroblasts was normal. The hypermethioninemia and a concurrently increased blood S-adenosylmethionine declined on a diet low in methionine. There was a diffuse, symmetrical, moderate proximal muscle weakness, but muscle atrophy was not discernible, and the deep tendon reflexes were hypoactive but obtainable. Electromyographic abnormalities were not detected. Electron microscopy of muscle revealed 3 to 6 small myelin figures in the region of the I band in nearly every fiber, with occasional myelin figures at other sites also. These myelin figures were more numerous and smaller than those seen accompanying nonspecific myopathies and may reflect a more specific pathological change. Electron microscopy of liver revealed three nonspecific lesions in all hepatocytes: (1) numerous megamitochondria with crystalloid deposit in the matrix; (2) increased numbers of small vesicles of smooth endoplasmic reticulum; and (3) loss of plasma membrane microvilli, with extensive bleb formation and shedding of cytoplasm into Disse's space.

Hypermethioninemia associated with methionine adenosyltransferase deficiency: clinical, morphologic, and biochemical observations on four patients.

Four patients with hypermethioninemia were ascertained in neonatal mass metabolic screening programs. Hypermethioninemia has persisted in all cases. There were no other abnormalities in sulfur-amino acid concentrations, and routine serum chemical determinations, including the results of "liver function" tests, were normal. Hepatic methionine adenosyltransferase activity was found to be low, ranging from 7.8 to 17.5% (mean 11.4%) of the normal adult control value. Electron microscopy of liver showed increased smooth endoplasmic reticulum, decreased rough endoplasmic reticulum, and increased lysosomes; short breaks in the outer membranes of mitochondria were present to a variable extent. Despite the persistent hypermethioninemia, which argues for continued deficiency of hepatic MAT, all four children appear well. This ostensible well being may be a result of the normal activity of extrahepatic MATs, as shown for erythrocytes and for cultured fibroblasts and lymphoid cells.

Mitochondrial abnormalities of liver in primary ornithine transcarbamylase deficiency.

Deficiency of hepatic ornithine transcarbamylase (EC 2.1.3.3) activity in a 17-month-old female patient is described. Enzyme activity was 11% of the mean control value. Electron microscopic examination of the liver specimen, taken by percutaneous needle biopsy, revealed striking abnormalities of the mitochondria: bud-like projections, sausage-link appearance, elongation with short cristae, or the presence of parallel arrays of tubules. The abnormalities do not resemble those seen in Reye's syndrome.

Cystathionase deficiency: the effect of cofactor on the stability of normal and abnormal enzyme from lymphoid cell lines.

Heating at 70 degrees C with and without added pyridoxal phosphate (PLP) had strikingly different effects on cystathionase protein from normal long-term lymphoid cell lines and on the enzymes from cells derived from patients with vitamin-B6-responsive cystathioninuria. PLP added to extracts of normal cells afforded complete protection against heat inactivation, whereas inactivation of the cystathionase protein in extracts obtained from two cystathioninuric lines was greater in the presence of PLP than in its absence.

Cystathionine beta-synthase deficiency: a qualitative abnormality of the deficient enzyme modified by vitamin B6 therapy.

The thermostability of cystathionine synthase and the effect of pyridoxal phosphate (PLP) on this thermostability were investigated in extracts of normal human liver and in extracts of liver, both before and during pyridoxine (vitamin B6) therapy, from members of a family with three clinically and biochemically typical, B6-responsive, synthase-deficient sibs. Incubation of crude extracts of normal liver at 55 degrees (preincubation) for 3-4 min before assay consistently resulted in a more than 2-fold increase in specific activity (activation) of cystathionine synthase (Fig. 1). With periods of preincubation longer than 4 min, thermal inactivation occurred. When PLP was added to the preincubation mixture, slightly more activation occurred in the first 3-4 min, and there was no observable loss of activity for an additional 25 min. The activation phenomenon was not observed in extracts of liver which had been obtained from three synthase-deficient sibs before therapy with vitamin B6 (Index of activation, Table 1). When extracts of liver obtained during vitamin B6 therapy were studied, however, significant activation was observed. Synthase activity in extracts of liver from the patients' parents, obligate heterozygotes for synthase deficiency, and from a potentially heterozygous sister demonstrated activation similar to that found in control liver extracts. With periods of preincubation longer than 5 min, the inactivation of synthase in liver extracts from patients receiving pyridoxine-HCl occurred at the same rate as in liver extracts from heterozygotes and from normal subjects (Index of inactivation, Table 1). PLP completely prevented heat inactivation of enzyme from normal liver.

Vitamin B6-responsive and -unresponsive cystathioninuria: two variant molecular forms.

Cystathionase activity in a lymphoid cell line extracts from a vitamin B6-responsive patient with cystathioninuria was increased strikingly by pyridoxal phosphate. Immunodiffusion with antiserum to human hepatic cystathionase showed identity between this cystathionase protein and cystathionase from an extract of normal lymphoid cells. Neither an increase in cystathionase activity nor immunochemical identity was found using extract of cells from a B6-unresponsive patient.

Methionine adenosyltransferase deficiency: new enzymatic defect associated with hypermethioninemia.

A specific deficiency of methionine adenosyltransferase has been demonstrated in the liver of an infant with hypermethioninemia. Since the enzymatic activity was below that in fetal liver and the metabolic abnormality has persisted (the infant now being 1 year of age), there is probably a genetic mutation. Mass screening for hypermethioninemia may uncover more such cases.