bluuebunny/crossref_metadata_2025 huggingface dataset:
Complementation studies with clinical and biochemical characterizations of a new variant of multiple sulphatase deficiency
Authors: Tanaka A. , Hirabayashi M. , Ishii M. , Yamaoka S. , Kawamura M. , Nishida M. , Isshiki G.
Published: June 2005

A patient with a new variant of multiple sulphatase deficiency (MSDv) is reported. Unlike the usual type, onset was late and progress was slow. The phenotypic changes were those usually seen in multiple sulphatase deficiency but much milder. Cytoplasmic accumulations were found in skin fibroblasts, and urinary mucopolysaccharides and sulphatides were high. Arylsulphatases A, B and C (ASA, B and C), heparan N‐sulphatase sulphoiduronate sulphatase, andN‐acetylgalactosamine 6‐sulphatase all had low activity in lymphocytes and cultured skin fibroblasts. Complementation for ASA activity was found in hybrids between MSDv and metachromatic leukodystrophy (MLD) as well as between multiple sulphatase deficiency (MSD) and MLD. Complementation for ASC activity was also seen in hybrids between MSDv and X‐linked ichthyosis (XLI), and between MSD and XLI. However, neither ASA nor ASC activity increased in hybrid cells of MSDv and MSD. These results suggested that the mutations of MSDv and of MSD were allelic, although of different phenotypes.
Elevation of serum β‐hexosaminidase and α‐d‐mannosidase in type 2 gaucher disease: a clinical and biochemical study
Authors: Chitayat D. , Nakagawa S. , Marion R. W. , Sachs G. S. , Shinnar S. , Llena J. F. , Nitowsky H. M.
Published: June 2005

We report a case of a black infant who died at 9 months of age with clinical and pathological findings consistent with the acute neuronopathic form of Gaucher disease (Type 2). Analysis of peripheral blood platelets obtained from this child demonstrated very low levels of β‐glucosidase activity. β‐hexosaminidase (HEX) activity in the serum, however, was 30 times greater than the level in control sera and 15 times greater than the level observed in individuals affected with the chronic form of Gaucher disease (Type 1). Similarly, α‐d‐mannosidase (MANN) activity in the proband's serum was significantly elevated when compared with controls, and chronic Gaucher disease patients. We postulate that the cause of the elevation of these lysosomal enzymes is similar to the cause of elevation in Type 1 individuals but that patients with Type 2 Gaucher disease have a more serious cellular defect.
3‐Hydroxy‐3‐methylglutaric aciduria: Response to carnitine therapy and fat and leucine restriction
Authors: Dasouki M. , Buchanan D. , Mercer N. , Gibson K. M. , Thoene J.
Published: June 2005

A female infant, born to first cousin parents, lapsed into coma with severe metabolic acidosis on day three of life. The gas chromatographic/mass spectrometric urinary organic acid profile showed marked elevation of the leucine metabolites 3‐hydroxy‐3‐methylglutaric, 3‐methylglutaconic, 3‐methylglutaric and 3‐hydroxy‐isovaleric acids. Less than 5% of the normal activity of the enzyme 3‐hydroxy‐3‐methylglutaryl CoA lyase was detected in cultured skin fibroblasts. The patient's total and free carnitine was initially low but rose to normal levels after placing her ondl‐carnitine (100 mg kg−1 d−1). On a diet providing 87 mg kg−1 d−1 of leucine and only 25% of total calories as fat and 2 g kg−1 d−1 protein, the concentration of the urinary organic acids fell markedly. She is now 15 months old with normal growth and development. This regimen appears effective in the early treatment of 3‐hydroxy‐3‐methylglutaric aciduria.
Haemodialysis for metabolic decompensation in propionic acidaemia
Authors: Roth B. , Younossi‐Hartenstein A. , Skopnik H. , Leonard J. V. , Lehnert W.
Published: June 2005

During an acute respiratory infection, a 4‐year‐old girl with propionic acidaemia deteriorated rapidly, becoming comatose with a marked acidosis. She did not respond to standard treatment but during haemodialysis her clinical state quickly improved.
Maternal PKU syndrome in an exceptional family with unexpected PKU
Authors: De Klerk J. B. C. , Wadman S. K. , Dijkhuis H. J. , Meuleman E. E. E.
Published: June 2005

Mild maternal phenylketonuria needs treatment during pregnancy.
Use of intact erythrocytes in the diagnosis of inherited purine and pyrimidine disorders
Authors: Fairbanks L. D. , Simmonds H. A. , Webster D. R.
Published: June 2005

This paper reports the detection of five inherited disorders of purine and one of pyrimidine metabolism using intact red blood cells (RBCs) and compares the findings with those from RBC lysate activity. Two different phosphate levels (1 and 18 mmol L−1 Pi) were used to evaluate endogenous PP‐ribose‐P levels and their generation by PP‐ribose‐P synthetase. The importance of this dual approach is demonstrated by the following evidence:

Six out of eight patients with no detectable hypoxanthine‐guanine phosphoribosyltransferase (HGPRT) RBC lysate activity had up to 25% of normal activity in their intact RBCs. Two Lesch‐Nyhan patients showed no detectable activity in intact or lysed RBCs.

RBC lysates from two heterozygotes for adenosine deaminase (ADA) deficiency also showed no detectable activity, but up to 60% of normal activity using intact RBCs.

The existence of an aberrant enzyme in a kindred with a superactive PP‐ribose‐P synthetase was evident from the fact that intact RBCs failed to respond normally to phosphate activation, despite normal HGPRT and adenine phosphoribosyltransferase (APRT) RBC lysate activity.

Raised endogenous PP‐ribose‐P levels in intact RBCs were demonstrable only in purine nucleoside phosphorylase (PNP) and HGPRT deficiency; levels were normal in APRT deficiency and hereditary oroticaciduria (OPRT/ODC) deficiency.

The results indicate that diagnosis from RBC lysate activity alone may be misleading. Intact RBC studies clearly provide a better indication of the functional capacity of the enzymein vivo. They also show a closer correlation with the clinical phenotype and allow further insight into the associated biochemical abnormalities in some cases.
Human β‐mannosidase deficiency: Biochemical findings in plasma, fibroblasts, white cells and urine
Authors: Cooper A. , Hatton C. , Thornley M. , Sardharwalla I. B.
Published: June 2005

Marked deficiencies of β‐mannosidase activity were demonstrated in plasma, leukocytes, fibroblasts and urine of a patient with β‐mannosidosis, similar deficiencies were observed in the proband's sibling. All other lysosomal enzymes measured, including sulphamidase, exhibited normal activity. Both parents showed reduced plasma and leukocyte β‐mannosidase activity. Urinary glycosaminoglycan excretion was normal but TLC of urinary oligosaccharides revealed an abnormal band with the mobility of a disaccharide. This finding was confirmed by Bio‐Gel P2 column chromatography. Further purification of this compound revealed two disaccharides, both of which yielded mannose and glucosamine following acid hydrolysis and mannose andN‐acetylglucosamine following enzymic digestion. These two compounds are thought to be structural isomers of the disaccharide Manβ‐GlcNAc.
3‐hydroxy‐3‐methylglutaryl‐coenzyme a lyase deficiency: Report of five new patients
Authors: Gibson K. M. , Breuer J. , Kaiser K. , Nyhan W. L. , McCoy E. E. , Ferreira P. , Greene C. L. , Blitzer M. G. , Shapira E. , Reverte F. , Conde C. , Bagnell P. , Cole D. E. C.
Published: June 2005

Five new patients are reported and the pathogenesis of the hypoglycaemia without ketogenesis is discussed. This report extends a recent review.J. Inher. Metab. Dis. 9, 225–233.
Cockayne syndrome: Magnetic resonance images of the brain in a severe form with early onset
Authors: Nishio H. , Kodama S. , Matsuo T. , Ichihashi M. , Ito H. , Fujiwara Y.
Published: June 2005

A 2‐year‐old boy with an early onset and severe form of Cockayne's syndrome (CS) showed differences from the common CS form, which made the clinical diagnosis difficult. However, the cellular characteristics of CS, that the patient's skin fibroblasts exhibited the hypersensitivity to the lethal effect of 254 nm ultraviolet light (UV) and a defective recovery of post‐UV DNA synthesis, but normal level of UV‐induced unscheduled DNA synthesis, were observed. The magnetic resonance imaging (MRI) by the inversion‐recovery method of the brain at age of 26 months showed atrophy or poor development of high signal images of the white matter. The MRI spin‐echo image showed a low signal image of the lenticular nucleus. The T1 and T2 values of the cerebrum (grey matter, white matter, lenticular nucleus and thalamus) were greater than those of the age‐matched controls, but similar to infant brains with much free water. Such MRI findings may suggest hypomyelination leading to the severe atrophy of the brain in this CS patient. His severe symptoms progressed rapidly until his death at 35 months due to systemic sepsis and renal dysfunction. Autopsy revealed severe microcephaly, severe atrophy of cerebrum, cerebellum and brain stem, and calcification throughout the brain, especially in the basal ganglia. Myelin staining showed numerous patchy losses of myelination in the cortical white matter.
The effects of fetal energy depletion on amniotic fluid concentrations of amino acids, organic acids and related metabolites
Authors: Harkness R. A. , Purkiss P. , Duffy S. , Chalmers R. A. , Jones M.
Published: June 2005

Concentrations of amino and organic acids, phosphate, sulphate, gluconic acid and gluconolactone were measured in amniotic fluid samples which contained either normal or raised hypoxanthine concentrations. In this way, the effect of mild fetal ATP depletion could be determined.

The effects of this mild asphyxia were to raise concentrations of phenylalanine, tyrosine, lysine, glycine, phosphate, sulphate, gluconic acid and glucono‐1,5‐lactone. However, concentrations of a variety of other metabolites were unchanged; thus no diagnostic confusion should arise with organic acidurias in mild asphyxia in contrast to the biochemical mimickry produced by severe asphyxia. Since clinically normal parturition can produce changes in amniotic fluid, urine from newborn or cord blood may not reflect the metabolic balancein utero.
Deficiency of UMP synthase in dairy cattle: A model for hereditary orotic aciduria
Authors: Harden K. K. , Robinson J. L.
Published: June 2005

Deficiency of uridine‐5′‐monophosphate (UMP) synthase in dairy cattle, a condition analogous to human hereditary orotic aciduria, is reviewed with consideration of similarities and differences between the enzyme deficiency in humans and cattle. New findings regarding the bovine condition are reported including presence of the enzyme deficiency in numerous tissues and absence of substantial effects on other aspects of nucleotide metabolism. Specifically, erythrocyte concentration of phosphoribosylpyrophosphate (PRPP) and activities of PRPP synthetase, adenine phosphoribosyltransferase, and hypoxanthine‐guanine phosphoribosyltransferase appear to be normal in cattle heterozygous for UMP synthase deficiency.
Studies on cross‐reacting material to steroid sulphatase in fibroblasts from patients affected by different types of steroid sulphatase deficiency
Authors: Parenti G. , Ballabio A. , Hoogeveen A. T. , Van Der Loos C. M. , Jobsis A. C. , Andria G.
Published: June 2005

Immunologically cross‐reacting material to antibodies against steroid sulphatase has not been found in fibroblasts from patients with steroid sulphatase deficiency.
Successful long term therapy of biopterin deficiency
Authors: Snyderman S. E. , Sansaricq C. , Pulmones M. T.
Published: June 2005

A hyperphenylalaninaemic infant, started on dietary therapy at 14 days of age, had severe developmental retardation and neurological abnormalities despite excellent biochemical control. A diagnosis of a deficit in biopterin synthesis was made at five months of age as a result of the following: high neopterin and low biopterin levels in both blood and urine, normal dihydropteridine reductase in the liver and a sharp drop in the plasma phenylalanine level 4 h after the administration of a test dose of tetrahydrobiopterin. Treatment with levodopa, carbidopa and 5‐hydroxytryptophan resulted in prompt neurological improvement. This was followed by gradual and sustained development. At present, at the age of 7 years, the child is mentally and physically normal.
Influence of the phenylketonuric heterozygote on the developing fetus
Authors: Cabalska B. , Miesowicz I. , Zorska K. , Nowaczewska I. , Duczynska N.
Published: June 2005

Physical development scores and clinical condition were evaluated in 403 newborn of phenylketonuric (PKU) heterozygotes. No significant differences were found between this group and the progeny of women from a healthy population. Analysis of mental development scores in 82 non‐PKU children age 3–14 years born to heterozygous mothers revealed normal values of psychometric tests.
Spectral studies of the interaction of the substrate ‘quinonoid’ 6‐methyl dihydropterine and the coenzyme NADH used as marker in the dihydropteridine reductase assay
Authors: van der Heiden C. , Brink W.
Published: June 2005

In dihydropteridine reductase assay the substrate quinonoid dihydropterine is reduced again to tetrahydropterine, concomitantly oxidizing NADH, the indicator of the enzyme assay. Because of the strong oxidizing capacity of quinonoid dihydropterine, the degree of spontaneous oxidation of NADH by the various substances used in the dihydropterine reductase assay was studied spectrally. A high degree of spontaneous oxidation of NADH by the substrate itself was found, which can be regulated by dithiotreitol, dependent on its concentration. The absorbance increase at 336 nm, due to the non‐quinonoid dihydropterine formed spontaneously from its quinonoid form, strongly interferes with the absorbance decrease at 340 nm, due to the disappearance of NADH. The interference results in a shift of the absorbance maximum of NADH from 340 nm up to higher wavelengths. This phenomenon, expressing itself in various ways in blank and sample, is discussed with relevance to the validity of the current enzyme assays used in a further classification of hyperphenylalaninaemic patients.
Methylmalonic acidaemia due to mutase apoenzyme defect: Responsive to vitamin B12 in intact fibroblasts but notin vivo
Authors: Baumgartner R. , Giardini O. , Cantani A. , Sabetta G. , Castro M.
Published: June 2005

Congenital methylmalonic acidaemia (MMA‐aemia) was diagnosed in an 8‐month‐old girl who presented with severe metabolic acidosis, hypoglycaemia and hyperglycinaemia. Vomiting, failure to thrive and apathy first appeared when breast feeding was replaced by a cows' milk formula at the age of 3 months. The patient, unresponsive to OH‐Cbl therapy, was successfully treated with dietary protein restriction and with Shohl's solution. Aged 4 years 9 months, she is in good health. Studies in cultured fibroblasts revealed a defect of the MMA‐CoA mutase apoenzyme. Mutase activity in cell extracts was barely detectable both with and without added coenzyme (Ado‐Cbl). Addition of OH‐Cbl to the culture medium improved overall propionate metabolism in intact fibroblasts but had no effect on mutase activity in cell extracts. These observations point to the presence of a very labile mutant enzyme, suggesting that the patient reported here may be suffering from yet another variant of MMA‐aemia.
Long‐term prognosis in galactosaemia: Results of a survey of 350 cases
Authors: Waggoner D. D. , Buist N. R. M. , Donnell G. N.
Published: June 2005

An international survey of the long term results of treating galactosaemia has shown poor results. These do not seem to be related to any of the relevant variables studied, for example delayed diagnosis or poor dietary compliance.
Culture of galactosaemic fibroblasts in the presence of galactose: Effect of inosine
Authors: Pourci M. L. , Mangeot M. , Soni T. , Lemonnier A.
Published: June 2005

Fibroblasts from three galactosaemics had no galactose‐1‐phosphate uridyltransferase (GALT) activity. These fibroblast cells were cultured in different media supplemented with dialysed fetal calf serum. Galactosaemic and control cell strains stopped growing in hexose‐free medium. In glucose‐free medium containing galactose, galatosaemic cells, in contrast to control cells, stopped growing after two days and died. In the same medium supplemented with inosine, they exhibited the same growth pattern as the control cell strains although in the presence of high concentrations of galactose‐1‐phosphate (Gal‐1‐P). These findings indicated that the glucose‐free medium containing galactose supplemented with dialysed fetal calf serum and inosine, as a ribose donor, was appropriate for furtherin vitro investigations of galactose metabolism in galactosaemic cells.
Characterization of human fructose‐1,6‐bisphosphatase in control and deficient tissues
Authors: Adams A. , Redden C. , Menahem S.
Published: June 2005

The regulatory properties of human liver and muscle fructose‐1, 6‐bisphosphatases (FBPase) have been studied in control tissues obtained at autopsy and in tissues from a neonate with FBPase deficiency who died as a result of an overwhelming acidosis. Evidence is presented which suggests that the alkaline isoenzyme of FBPase, which is widely regarded as a laboratory artefact, may have an important rolein vivo in the regulation and control of glycolysis and gluconeogenesis. FBPase exhibits the hysteretic and dissociative properties associated with regulatory enzymes, and many of the factors which effect FBPase have inverse effects on phosphofructokinase activity, thus providing an integrated regulatory cycle for the control of the direction and rate of flux through the glycolytic pathway.
Glycogen debranching enzyme deficiency: Long‐term study of serum enzyme activities and clinical features
Authors: Coleman R. A. , Winter H. S. , Wolf B. , Chen Y. ‐T.
Published: June 2005

In glycogen storage disease type III (glycogen debranching enzyme (DE) deficiency), the activities of serum alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase may be strikingly elevated during childhood but are low during adult life. To determine the pattern of the elevated serum enzyme activities in relationship to diet, the biochemical subtype and clinical symptoms, 13 patients with DE deficiency were studied. Activities of serum aspartate and alanine transaminases, lactate dehydrogenase, and alkaline phosphatase were markedly elevated during infancy. Continued elevation of enzyme activities during childhood appeared to be related to DE deficiency in liver, but unrelated to DE deficiency in muscle. Activity elevations correlated inconsistently with diet and poorly with childhood growth rate or the presence of hypoglycaemia. The serum enzyme activities declined around puberty concomitantly with a decrease in liver size. Although periportal fibrosis and micronodular cirrhosis indicated the presence of hepatocellular damage during childhood, the decline in serum enzyme activities with age and the absence of overt hepatic dysfunction suggest that the fibrotic process may not always progress.
Tryptophan and serotonin metabolism in familial erythrophagocytic lymphohistiocytosis
Authors: Howells D. W. , Hyland K. , Smith I. , Strobel S.
Published: June 2005

Reduced concentrations of tryptophan and 5‐hydroxyindoleacetic acid (the major CSF metabolite of serotonin) were found in the cerebrospinal fluid of two children with familial erythrophagocytic lymphohistiocytosis. This was associated with elevated cerebrospinal fluid neopterin concentrations indicating increased macrophage activity within the central nervous system. In one child, cytotoxic therapy induced a complete clinical remission and an increase of tryptophan and 5‐hydroxyindoleacetic acid concentrations to normal; during a subsequent relapse, concentrations of these analytes again fell below normal. In the other child, in whom therapy produced only a transient improvement, tryptophan and 5‐hydroxyindoleacetic acid concentrations remained low and the child died. It is likely that increased activity of indoleamine 2,3‐dioxygenase induced by the activation of macrophages was responsible for the disturbance in serotonin and tryptophan homeostasis within the brain. Excessive tryptophan catabolism and the disturbance of serotonin turnover may play a role in the aetiology of the neurological symptoms seen in familial erythrophagocytic lymphohistiocytosis.
Attempted enzyme replacement using human amnion membane implantations in mucopolysaccharidoses
Authors: Muenzer J. , Neufeld E. F. , Constantopoulos G. , Caruso R. C. , Kaiser‐Kupfer M. I. , Pikus A. , Danoff J. , Berry R. R. , McDonald H. D. , Thompson J. N. , Rodén L. , Zasloff M. A.
Published: June 2005

Amnion membrane implantation has been proposed as an approach to enzyme replacement in mucopolysaccharidoses. Human amnion membranes have been subcutaneously implanted in the abdominal wall in 19 patients with mucopolysaccharidoses (MPS I, II and III). A protocol was developed for the objective evaluation of experimental treatments of these patients. Systematic evaluation of the clinical status before and 6 months after amnion membrane implantation reveals no change in function except improvement in joint mobility. The sum of all joint movements showed improvement from baseline values to 6 months after implantation by ANOVA followed bypost‐hoc analysis (p<0.056). The only specific joint movements to significantly improve after 6 months were shoulder extension (p<0.01) and hip internal rotation (p<0.05). Serial measurements of the deficient lysosomal enzyme activity in serum and white blood cells did not increase in any patient after amnion membrane implantation. Urinary glycosaminoglycan excretion decreased transiently in 2 of 10 patients after implantation, but a second amnion membrane implantation did not result in any change. Biopsy of the implantation site in 10 patients 6 months after amnion membrane implantation revealed a foreign‐body reaction with giant cell formation and fibrosis and no recognizable amnion membrane tissue. We conclude that human amnion membrane implantation is not an effective therapy in mucopolysaccharidoses.
Oligosaccharides accumulated in the bovineβ‐mannosidosis kidney
Authors: Jones M. Z. , Rathke E. J. S. , Gage D. A. , Costello C. E. , Murakami K. , Ohta M. , Matsuura F.
Published: June 2005

The phenotype of bovineβ‐mannosidosis (β‐mannosidase deficiency), recently identified in Salers cattle, is similar to the caprine form of the disease (Abbittet al., 1991). This investigation was designed to characterize accumulated kidney oligosaccharides in bovineβ‐mannosidosis. Oligosaccharides were extracted from the kidney of an affected Salers calf and purified by chromatographic techniques. The amount of accumulating oligosaccharides in 1 g of wet tissue was about 21µmol. Structures of derivatized oligosaccharides were characterized by high‐performance liquid chromatography, mass spectrometry, methylation analysis and sequential exoglycosidase digestions. The major accumulating oligosaccharides were Manβ1‐4GlcNAc and Manβ1‐4GlcNAcβ1‐4GlcNAc. Oligosaccharides accumulating in minor amounts were Manβ1‐4GlcNAcβ1‐4Manβ1‐4GlcNAc, Manα1‐6Manβ1‐4GlcNAcβ1‐4GlcNAc and Manβ1‐4GlcNAcβ1‐4Manβ1‐4GlcNAcβ1‐4GlcNAc. As in caprineβ‐mannosidosis, oligosaccharides with terminalβ‐mannose residues and cleaved as well as uncleaved chitobiose linkages were identified in bovineβ‐mannosidosis kidney. The accumulating oligosaccharides in tissue were thus identical in bovine and caprineβ‐mannosidosis; however, the source of the novel oligosaccharides remains to be determined.
Gonadal mosaicism in a family with adrenoleukodystrophy: Molecular diagnosis of carrier status among daughters of a gonadal mosaic when direct detection of the mutation is not possible
Authors: Graham G. E. , MacLeod P. M. , Lillicrap D. P. , Bridge P. J.
Published: June 2005

Adrenoleukodystrophy is a severe, X‐linked neurological disease that has been shown to be linked to DNA markers from Xq28. We tested several families with these markers and, in one family, found two apparent recombination events between DXS52 and the disease. Expansion of the study to include other tests and several others markers from Xq28 led us to conclude that recombination probably had not occurred and that, instead, the mutation in this family had a mitotic origin and that the grandmother was a gonadal mosaic. For genes that have been cloned, it is often possible to demonstrate the presence or absence of a specific mutation in such families and to determine carrier status on that basis. This is not possible when the gene has not been cloned. We therefore describe a method that can be employed by a molecular diagnostic laboratory to discriminate between people who inherit the same RFLP halpotype, with or without the mutation, from a parent with gonadal mosaicism in diseases where direct gene analysis is not yet possible.
Quantification ofN‐acetyl‐l‐aspartic acid in urine by isotope dilution gas chromatography‐mass spectrometry
Authors: Kelley R. I. , Stamas J. N.
Published: June 2005

A method for quantification ofN‐acetyl‐l‐aspartic acid by isotope dilution gas chromatography‐mass spectrometry using15N‐[2H3]acetyl‐l‐aspartic acid is described. The method is sufficiently sensitive to be used with solvent extraction techniques commonly employed for urinary organic acid analysis. The mean concentration ofN‐acetyl‐l‐aspartic acid in 80 normal and abnormal control urine specimens was 19.7±10.8 mg/g creatinine (12.7±7.8 mmol/mol creatinine). Seven patients, ages 9 months to 7 years, with Canavan‐van Bogaert syndrome had urinaryN‐acetyl‐l‐aspartic acid levels from 606 to 4760 mg/g creatinine. The method can also be used with cerebrospinal fluid, in which the concentration ofN‐acetyl‐l‐aspartic acid is about one‐tenth of that in urine.
Biochemical basis of hypoxanthine‐guanine phosphoribosyltransferase deficiency in nine families
Authors: Keough D. T. , Gordon R. B. , de Jersey J. , Emmerson B. T.
Published: June 2005

The concentration of hypoxanthine‐guanine phosphoribosyltransferase (HPRT) cross‐reacting material (CRM) was determined in haemolysates and/or lymphoblast lysates from nine patients with complete or partial deficiency of HPRT activity. Two of the patients had the fully developed Lesch‐Nyhan syndrome and although they had undetectable HPRT activity, small amounts of CRM were found. HPRT‐specific mRNA was not detected in lymphoblast lysates from one of these patients, while lysates from the other had a much reduced concentration. Samples from three patients with <0.1% of normal HPRT activity but with minor or no neurological manifestations were also found to contain small amounts of CRM. The other four patients whose HPRT activities ranged from 3 to 10% of normal were found to have CRM concentrations which varied from 26 to 100% of normal. In one patient with a partial deficiency theKm for 5‐phospho‐α‐d‐ribosyl‐1‐pyrophosphate was five times normal.
Use of platelets, mononuclear and polymorphonuclear cells in the diagnosis of glycogen storage disease type VI
Authors: Dahan N. , Baussan C. , Moatti N. , Lemonnier A.
Published: June 2005

We determined glycogen concentration and phosphorylase ‘a+b’ and phosphorylase a activities in platelets, mononuclear and polymorphonuclear cells from control subjects and patients with phosphorylase kinase deficiency (glycogen storage disease IX) and liver phosphorylase deficiency (glycogen storage disease VI). Variations according to cellular type and to subjects' age (1–40 years) were established. Variable glycogen overloading was found in all our patients. Glycogen storage disease (GSD) VI was characterized by a diminished total phosphorylase activity with a low or normal a/(a+b) ratio of phosphorylase activity. GSD IX was characterized by a very low residual activity of phosphorylase a with an ‘a+b’ activity low or normal.
Excretion of phenylpyruvic, 4‐hydroxyphenylpyruvic and indolyl‐3‐acetic acids by the skin fibroblasts from a phenylketonuric child
Authors: Antoshechkin A. G. , Zuyeva L. A. , Maximova L. A.
Published: June 2005

High‐resolution gas chromatography and mass spectrometry have been used to analyse the changing chemical composition of the culture medium upon the growth of skin fibroblasts from a healthy individual and from a classical phenylketonuric (PKU) patient. The PKU fibroblasts, unlike normal ones, were found to excrete into the culture medium phenylpyruvate, 4‐hydroxyphenylpyruvate, indolyl‐3‐acetate and an unidentified metabolite containing the phenyl group. This testifies to the disturbed metabolism of phenylalanine, tyrosine and tryptophan in the PKU fibroblasts. This seems to be the reason for their reduced viability as compared with normal fibroblasts.
Peroxisomal integral membrane proteins in livers of patients with Zellweger syndrome, infantile Refsum's disease and X‐linked adrenoleukodystrophy
Authors: Small G. M. , Santos M. J. , Imanaka T. , Poulos A. , Danks D. M. , Moser H. W. , Lazarow P. B.
Published: June 2005

Livers from seven patients with peroxisome disorders, three with Zellweger syndrome, one with infantile Refsum's syndrome and three with X‐linked adrenoleukodystrophy, were analysed by immunoblotting. The bifunctional protein catalysing two peroxisomal β‐oxidation reactions was deficient in all Zellweger livers and in the infantile Refsum's liver, consistent with the absence of morphologically recognizable peroxisomes. Three peroxisomal integral membrane proteins (IMPs) (69, 53 and 22 kDa) were present in normal amounts in all the Zellweger and adrenoleukodystrophy samples and they sedimented in a membrane fraction. These membrane proteins were also present in the infantile Refsum's liver. We suggest, on the basis of these results, that aberrant peroxisomal membranes may be present in Zellweger syndrome and that the defect is in the transport of matrix proteins into the organelle.
Polarizing inclusions in some organs of children with congenital peroxisomal diseases (Zellweger's, Refsum's, chondrodysplasia punctata (rhizomelic form), X‐linked adrenoleukodystrophy)
Authors: Kerckaert I. , Dingemans K. P. , Heymans H. S. A. , Vamecq J. , Roels F.
Published: June 2005

Polarizing material has been reported in the liver of children with infantile Refsum's disease (IRD) and was absent in two patients with the cerebro‐hepato‐renal syndrome of Zellweger (CHRS). We examined in polarized light 15 liver biopsy and autopsy samples from six other patients with the cerebro‐hepato‐renal syndrome of Zellweger, two with the rhizomelic form of chondrodysplasia punctata (rCDP) and two with X‐linked adrenoleukodystrophy (ALD), all conditions with deficient peroxisomes. Two types of birefringent inclusions were found in CHRS only: the first is transparent in bright field, the second appears as brown granules or rods, similar to lipofuscins. As in IRD large PAS‐positive macrophage‐like cells contain the transparent type. Electron microscopical investigation of these cells shows trilaminar structures within membrane‐bound organelles. The two types were also seen in kidney and brown adipose tissue, the first type in pancreas, the second type in adrenal gland; no such was observed in myocardium or in thyroid gland (CHRS). No birefringent inclusions were present in rCDP and ALD. The nature of the inclusions is still unclear. An accumulation of the transparent polarizing material with increasing age of the patients is most likely.
Metabolism of3H‐dehydroepiandrosterone sulphate by subjects with steroid sulphatase deficiency
Authors: Bergner E. A. , Shapiro L. J.
Published: June 2005

Patients with steroid sulphatase deficiency develop ichthyosis with accumulation of cholesterol sulphate in plasma and in the stratum corneum. The present study was undertaken to determine whether desulphation of the C19 steroid DHEAS is also impaired. The mean plasma concentrations of DHEA and androstenedione were significantly lower for patients than for controls (p<0.02 and 0.001) while the mean concentration of DHEAS was higher (p<0.002). Following intravenous administration of3H‐DHEAS, one patient failed to desulphate3H‐DHEAS as evidenced by an absence of urinary3H‐glucuronides. A second produced normal amounts of urinary3H‐glucuronides (indicative of desulphation capacity) in a baseline study but did not desulphate3H‐DHEAS following ampicillin treatment to alter gut microflora. A third patient had consistent sulphatase activity with and without ampicillin.
Pattern reversal visual evoked potentials in phenylketonuria
Authors: Giovannini M. , Valsasina R. , Villani R. , Ducati A. , Riva E. , Landi A. , Longhi R.
Published: June 2005

The pathogenesis of brain dysfunction in phenylketonuria (PKU) is still under investigation. Hyperphenylalaninaemia results in increased turnover of myelin. In order to demonstrate the derangement of myelinization in PKU we studied the visual evoked potentials (VEP) in 14 PKU patients and in 20 normal subjects. VEP findings were correlated with the metabolic control of the disease and with the electroencephalographic findings. VEP were more sensitive than the EEG in detecting a neurological dysfunction. VEP are influenced by dietary control and are normal only in children with good metabolic control.
Dyggve‐Melchior‐Clausen syndrome with increased pipecolic acid in plasma and urine
Authors: Roesel R. A. , Carroll J. E. , Rizzo W. B. , van der Zalm T. , Hahn D. A.
Published: June 2005

A child with the Dyggve‐Melchior‐Clausen syndrome associated with elevated pipecolic acid levels in plasma and urine is described. Other studies of peroxisomal function, including phytanic acid, very long‐chain fatty acids, and plasmalogen synthesis, were normal. This disorder may represent an incompletely characterized defect in peroxisomal metabolism.
Genetic expression of Menkes disease in cultured astrocytes of the macular mouse
Authors: Kodama H. , Meguro Y. , Abe T. , Rayner M. H. , Suzuki K. T. , Kobayashi S. , Nishimura M.
Published: June 2005

The copper concentration was investigated in the cultured astrocytes from macular mice, an animal model of Menkes disease. An excessive amount of copper was accumulated in the astrocytes as copper‐metallothionein. These results show that the underlying genetic defect of the macular mouse is expressed in the astrocytes. A similar situation may exist in Menkes disease and cause a failure of copper transport to neurones.
Worldwide survey of neonatal screening for biotinidase deficiency
Authors: Wolf B.
Published: June 2005

Neonatal screening for biotinidase deficiency has been conducted in 14 countries since 1984. To 31 December 1990, 8 532 617 newborns were screened. One hundred and forty‐two infants with biotinidase deficiency were identified; 76 infants with profound deficiency (<10% of mean normal serum activity) and 66 infants with partial deficiency (10–30% of mean normal activity). The estimated incidence of profound biotinidase deficiency is 1:112 271 (1:85 000 to 1:145 000; 95% confidence limits) and the incidence of partial deficiency is 1:129 282 (1:112 700 to 1:177 000). The incidence of combined profound and partial deficiency is 1:60 089 newborns (1:49 500 to 1:73 100). The estimated frequency of the allele for biotinidase deficiency is 0.004 and an estimated 1 in 123 individuals is heterozygous for the disorder.
Neonatal screening for biotinidase deficiency in East‐Hungary
Authors: Havass Z.
Published: June 2005

There are two types of multiple carboxylase deficiency, the neonatal form with holocarboxylase synthetase defect and the late‐onset form with biotinidase deficiency. We report our preliminary experiences in screening for biotinidase deficiency. In total 43 493 infants were screened for the deficiency of the enzyme biotinidase; 0.14% false positive results that necessitated requests for second blood samples and two newborns with a biotinidase defect were identified during our pilot study. The definitive diagnosis required the demonstration of enzyme deficiency in serum. Both of the patients have residual biotinidase activity: 3.59% and 7.55%. These two newborns with biotinidase deficiency are treated with daily supplementation of free biotin. According to our preliminary results biotinidase deficiency satisfies all the criteria for incorporation into the national newborn mass screening.
Combined xanthine and sulphite oxidase defect due to a deficiency of molybdenum cofactor
Authors: Roesel R. A. , Bowyer F. , Blankenship P. R. , Hommes F. A.
Published: June 2005

Increased urinary excretion of xanthine, hypoxanthine, sulphite, thiosulphate and decreased serum uric acid were observed in an infant with profound failure to thrive. Other clinical findings included refractory seizures, spastic quadriplegia and profound psychomotor retardation. The patient died at 20 months of age. There were no detectable activities for xanthine oxidase and sulphite oxidase in the postmortem liver. Urothione, which is the metabolic excretory product of the molybdenum cofactor for molybdoenzymes was not present in the urine. A deficiency of the molybdenum cofactor which is common to both xanthine and sulphite oxidase is presumed to be the metabolic defect responsible for the absent activities of both enzymes.
Identification of cystic fibrosis homozygotes and heterozygotes by isoelectric focussing of serum proteins
Authors: Getliffe K. A. , Ward L. C. , Shepherd R. W.
Published: June 2005

Screening of 44 cystic fibrosis homozygotes, 17 heterozygotes and 36 normal controls, by identification of cystic fibrosis protein (CFP), was performed on sera using an improved isoelectric focussing technique. CFP was observed in 91% of homozygotes, 88% of heterozygotes and 8% of normal controls tested. Partial purification of CFP by chromatofocussing indicates that CFP has a molecular weight of about 52000u. It is speculated that CFP is a normal serum protein that exhibits a quantitative difference in concentration between CF homozygotes, heterozygotes and normals.
Phosphoglycerate kinase: Studies on normal and a mutant human enzyme
Authors: Svirklys L. G. , Lee C. S. , O'Sullivan W. J.
Published: June 2005

Phosphoglycerate kinase was purified from a number of tissues obtained at autopsy from a subject with the deficiency. Properties of the mutant enzyme were compared to those of PGK purified from tissue obtained from normal subjects. The purified enzyme from the propositus contained two components, a minor fraction (about 2–5%) which behaved similarly to the normal enzyme during the purification procedure, and a major fraction (>95%) which could not be purified by the same procedure. The major fraction demonstrated a number of other properties which differed from the normal. These included a tendency to form aggregates, increased heat sensitivity and altered nucleotide substrate specificity. The smaller fraction appeared to have effectively identical properties to that of the normal enzyme. In keeping with its X‐linked mode of inheritance, phosphoglycerate kinase from all normal tissues appeared to have identical kinetic properties, although evidence for minor variations, presumably due to post‐translational modifications, was obtained.
Clinical consequences of hyperlipidaemia
Authors: Thompson G. R.
Published: June 2005

Hyperlipidaemia is a common and important clinical entity which frequently has a genetic basis. The chief features of severe hypertriglyceridaemia are eruptive xanthomata and acute pancreatitis, whereas most forms of hypercholesterolaemia are associated with premature coronary heart disease. This applies especially to familial hypercholesterolaemia, which is also characterized by tendon xanthomata. Better recognition of hyperlipidaemia allied to recent improvements in treatment should help reduce the frequency of its disabling and sometimes fatal consequences.
Apolipoprotein polymorphism and multifactorial hyperlipidaemia
Authors: Utermann G.
Published: June 2005

Apolipoproteins AIV, B, E, and the Lp(a) glycoprotein are genetically polymorphic in humans. Three common allelesɛ2,ɛ2 andɛ4 control the polymorphism of apolipoprotein E. These code for proteins which differ in functional properties, e.g. receptor binding activity andin vivo catabolism. This explains the significant effect of the apoE gene locus on the variability of plasma lipoprotein concentrations and moreover the implication of apoE alleles in the aetiology of multifactorial forms of hyperlipidaemia e.g. familial type III hyperlipidaemia (apoE2; arg158→cys) and polygenic hypercholesterolaemia (apoE4; cys112→arg). A further gene locus controls the concentrations in plasma of the Lp(a) lipoprotein that is composed of an LDL‐like particle containing apoB‐100 and the disulphide‐bonded Lp(a) glycoprotein. The latter exhibits a genetic size polymorphism (MW∼400kD‐700kD) that is controlled by at least seven autosomal alleles. These alleles at the same time are involved in determining the plasma concentrations of the lipoprotein that range from <1 mg/dl to >200mg/dl. Thus there is evidence that genetic variability in apolipoproteins relates to the variability of lipoprotein concentrations in the population and is implicated in the aetiology of multifactorial hyperlipidaemias.
Strategy for the screening of tetrahydrobiopterin deficiency among hyperphenylalaninaemic patients: 15‐Years experience
Authors: Dhondt J. ‐L.
Published: June 2005

Tetrahydrobiopterin deficiency in hyperphenylalaninaemic babies has to be rapidly recognized since the disease requires a specific treatment. Based on 15 years experience, we report on the evolution of a strategy for the detection of such patients. A total of 913 hyperphenylalaninaemic patients have been studied and 15 tetrahydrobiopterin deficiences have been detected or confirmed.

DHPR assay in dried blood samples and pteridine measurement in urine collected on filter paper combine convenient sampling and reliable tests for systematic investigation of hyperphenylalaninaemic patients for cofactor deficiency.
First trimester diagnosis of Inherited Metabolic Disease: Experience in the UK
Authors: Besley G. T. N. , Young E. P. , Fensom A. H. , Cooper A.
Published: June 2005

Experience with first trimester diagnosis of inherited metabolic disease is still limited. In this report, data are collected from four major centres in the UK which provide a prenatal diagnosis service based on specific enzyme or gene product assay. The data were presented at a workshop on ‘First Trimester Diagnosis of Inherited Metabolic Disease’ held at the Institute of Child Health, London, on 21st June 1990.

Approximately 100 different metabolic diseases can now be diagnosed in the first trimester, but because of the rarity of many of the disorders, experience of positive diagnoses, based on enzyme deficiency in fresh chorionic villus samples (CVS), cultured villus cells or early amniocentesis samples, is likely to be limited. It is, however, important that these results are reported and similarly that any problems which arise are fully documented.
Use of 4‐trifluoromethylumbelliferyl‐α‐l‐iduronide as a new substrate for detection of α‐l‐iduronidase deficiency in human tissues and for rapid prenatal diagnosis of Hurler disease
Authors: Tsvetkova I. V. , Karpova E. A. , Voznyi Y. V. , Zolotukhina T. V. , Biryukov V. V. , Semyachkina A. N.
Published: June 2005

Results are presented of α‐l‐iduronidase assays in the leukocytes of normal individuals, patients with Hurler disease and heterozygous carriers. The assays were carried out using 4‐methylumbelliferyl‐α‐l‐iduronide and 4‐trifluoromethylumbelliferyl‐α‐l‐iduronide as substrates. It was shown that 4‐trifluoromethylumbelliferyl‐α‐l‐iduronide, along with the commonly used 4‐methylumbelliferyl‐α‐l‐iduronide, can serve as a specific substrate for α‐l‐iduronidase and is therefore suitable for demonstrating the enzyme deficiency in patients with Hurler disease, as well as the decrease of enzyme activity in heterozygous disease carriers. Using the two substrates a prenatal diagnosis of Hurler disease in a fetus was made on the basis of the lack of enzyme activity in amniotic fluid cell cultures. The diagnosis was confirmed by the results of α‐l‐iduronidase activity assay in fetal liver and kidney. It was found that 4‐trifluoromethylumbelliferyl‐α‐l‐iduronide is highly efficient for the rapid detection of α‐l‐iduronidase deficiency directly in pieces of tissues and in placenta, which is important for the prenatal diagnosis of Hurler disease.
Characterization of the mutations in three patients with pyruvate dehydrogenase E1α deficiency
Authors: Hansen L. L. , Brown G. K. , Kirby D. M. , Dahl H. ‐H. M.
Published: June 2005

The human pyruvate dehydrogenase complex catalyses the oxidative decarboxylation of pyruvate to acetyl‐CoA. Defects in several of the seven subunits have been reported, but the majority of mutations affect the E1 component and especially the E1α subunit. However, the clinical presentation of patients with pyruvate dehydrogenase E1α deficiency is extremely variable. Dependency of the brain on pyruvate dehydrogenase activity and localization of the gene for the somatic form of the pyruvate dehydrogenase E1α subunit to the X chromosome provide the basis for a better understanding of the variation in the clinical manifestations. Further understanding of the function and interaction of subunits and the pathophysiology of pyruvate dehydrogenase deficiency necessitates the characterization of mutations in the pyruvate dehydrogenase complex. We report the analysis of three patients with pyruvate dehydrogenase E1α deficiency. One female has a three base pair deletion which affects dephosphorylation of the subunit. Of two males analysed, one has a two base pair deletion causing a shift in the reading frame. The other has a base change, resulting in an Arg to His substitution. All three mutations are located near the carboxyl terminus of the subunit.
Acyl‐CoA oxidase, peroxisomal thiolase and dihydroxyacetone phosphate acyltransferase: Aberrant subcellular localization in Zellweger syndrome
Authors: van Roermund C. W. T. , Brul S. , Tager J. M. , Schutgens R. B. H. , Wanders R. J. A.
Published: June 2005

We have studied the presence and subcellular localization of peroxisomal 3‐oxoacylcoenzyme A thiolase, acylcoenzyme A oxidase and acyl‐CoA: dihydroxyacetonephosphate acyltransferase (DHAPAT) in fibroblasts from control subjects and patients with an inherited deficiency of peroxisomes (Zellweger syndrome), using immunofluorescence spectroscopy and density gradient centrifugation techniques. The results show that Zellweger cells contain unprocessed thiolase and unprocessed acyl‐CoA oxidase which are associated with structures containing a peroxisomal integral membrane protein of 69 kDa and having a density much lower than that of normal peroxisomes. The residual DHAPAT activity present in Zellweger cells is also contained in these structures. We conclude that these structures represent defectively assembled peroxisomes which may still be capable of importing some peroxisomal proteins.
Atypical riboflavin‐responsive glutaric aciduria, and deficient peroxisomal glutaryl‐CoA oxidase activity: a new peroxisomal disorder
Authors: Bennett M. J. , Pollitt R. J. , Goodman S. I. , Hale D. E. , Vamecq J.
Published: June 2005

Investigation of cultured skin fibroblasts in a patient with atypical riboflavin‐responsive glutaric acidura revealed a marked deficiency of peroxisomal glutaryl‐CoA oxidase. This is the first patient to be reported with glutaric aciduria caused by a peroxisomal rather than a mitochondrial dysfunction. This enzyme appears to be specific for glutaryl‐CoA, as lauryl‐CoA and dodecanedioyl‐CoA oxidase activities in the fibroblasts were both normal. The urinary excretion of glutaric acid (0.5 mmol mmol creatinine−1) suggests that the flux through this pathway is considerably less than the mitochondrial flux through glutaryl‐CoA dehydrogenase. The elevated glutaric acid excretion (to 0.8 mmol mmol creatinine−1) in response to lysine loading suggests that lysine is a precursor.