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1.
Mol Genet Metab ; 106(3): 287-300, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22658377

ABSTRACT

Pompe disease is a genetic disorder resulting from a deficiency of lysosomal acid alpha-glucosidase (GAA) that manifests as a clinical spectrum with regard to symptom severity and rate of progression. In this study, we used microarrays to examine gene expression from the muscle of two cohorts of infantile-onset Pompe patients to identify transcriptional differences that may contribute to the disease phenotype. We found strong similarities among the gene expression profiles generated from biceps and quadriceps, and identified a number of signaling pathways altered in both cohorts. We also found that infantile-onset Pompe patient muscle had a gene expression pattern characteristic of immature or regenerating muscle, and exhibited many transcriptional markers of inflammation, despite having few overt signs of inflammatory infiltrate. Further, we identified genes exhibiting correlation between expression at baseline and response to therapy. This combined dataset can serve as a foundation for biological discovery and biomarker development to improve the treatment of Pompe disease.


Subject(s)
Glycogen Storage Disease Type II/genetics , Transcription, Genetic , alpha-Glucosidases/genetics , Age of Onset , Child , Child, Preschool , Female , Gene Expression , Glycogen Storage Disease Type II/metabolism , Humans , Infant , Infant, Newborn , Male , Muscle, Skeletal/metabolism , Phenotype , alpha-Glucosidases/metabolism
2.
Neurology ; 68(2): 110-5, 2007 Jan 09.
Article in English | MEDLINE | ID: mdl-17210890

ABSTRACT

BACKGROUND: Pompe disease (acid maltase deficiency, glycogen storage disease type II; OMIM 232300) is an autosomal recessive lysosomal storage disorder characterized by acid alpha-glucosidase deficiency due to mutations in the GAA gene. Progressive skeletal muscle weakness affects motor and respiratory functions and is typical for all forms of Pompe disease. Cardiac hypertrophy is an additional fatal symptom in the classic infantile subtype. c.-32-13T-->G is the most common mutation in adults. OBJECTIVE: To delineate the disease variation among patients with this mutation and to define the c.-32-13T-->G haplotypes in search for genotype-phenotype correlations. METHODS: We studied 98 compound heterozygotes with a fully deleterious mutation (11 novel mutations are described) and the common c.-32-13T-->G mutation. RESULTS: All patients were Caucasian. None had the classic infantile form of Pompe disease. The clinical course varied far more than anticipated (age at diagnosis <1 to 78 years; age at onset: <1 to 52 years). The acid alpha-glucosidase activities in a subset of patients ranged from 4 to 19.9 nmol/mg/h. Twelve different c.-32-13T-->G haplotypes were identified based on 17 single-nucleotide polymorphisms located in the GAA gene. In 76% of the cases, c.-32-13T-->G was encountered in the second most common GAA core haplotype (DHRGEVVT). In only one case was c.-32-13T-->G encountered in the major GAA core haplotype (DRHGEIVT). CONCLUSION: Patients with the same c.-32-13T-->G haplotype (c.q. GAA genotype) may manifest first symptoms at different ages, indicating that secondary factors may substantially influence the clinical course of patients with this mutation.


Subject(s)
Genetic Predisposition to Disease/genetics , Glycogen Storage Disease Type II/epidemiology , Glycogen Storage Disease Type II/genetics , Haplotypes/genetics , Risk Assessment/methods , alpha-Glucosidases/genetics , Adolescent , Adult , Aged , Child , Child, Preschool , Cohort Studies , DNA Mutational Analysis , Female , Glycogen Storage Disease Type II/enzymology , Humans , Infant , Infant, Newborn , Internationality , Male , Middle Aged , Mutation , Prevalence
4.
Mol Genet Metab ; 86(4): 466-72, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16185907

ABSTRACT

Gaucher disease is the most common of the lysosomal storage disorders, affecting all ethnic groups. The pathology of this recessively inherited disease arises from the accumulation of glucocerebroside in tissues due to deficient activity of the enzyme glucocerebrosidase (E.C. 3.2.1.45). The glucocerebrosidase (GBA) gene spans a 7.2kb fragment located on locus 1q 21, consisting of 11 exons and 10 introns. Located 16 kb downstream is a highly homologous pseudogene sequence [M. Horowitz, S. Wilder, Z. Horowitz, O. Reiner, T. Gelbart, E. Beutler, The Human Glucocerebrosidase gene and pseudogene: structure and evolution. Genomics 4 (1) (1989) 87-96.]. Fourteen fragments comprising 11 exons of the GBA gene were analyzed in DNA samples from 25 Colombian patients using denaturing High Pressure Liquid Chromatography (DHPLC). Sequencing of abnormal findings led to the discovery of three novel mutations (c.595_596 delCT, c.898 delG and c.1,255 G>C [p.D 419 H] in exons 6, 7, and 9 of the GBA gene) with high prevalence among Colombian patients. We have also found the presence of a double mutation p.L 483 P+p.E 355 K (L 444 P+E 326 K, traditional nomenclature) in two different families classified as Gaucher type 1. This mutation was previously reported in one patient with Gaucher type 2. We have found DHPLC to be a reliable and sensitive method for the detection of mutations and allelic variation in Gaucher patients.


Subject(s)
Gaucher Disease/enzymology , Gaucher Disease/genetics , Glucosylceramidase/genetics , Hispanic or Latino/genetics , Mutation , Adolescent , Adult , Base Sequence , Child , Child, Preschool , Colombia , DNA/genetics , DNA Mutational Analysis , Glucosylceramidase/deficiency , Humans , Middle Aged , Mutation, Missense , Sequence Deletion
5.
J Pharm Biomed Anal ; 35(2): 267-75, 2004 Apr 16.
Article in English | MEDLINE | ID: mdl-15063461

ABSTRACT

Nicardipine (NC)-cyclodextrin solid systems were prepared in equimolar ratios and their photostability in aqueous solution under exposure to UV(A)-UV(B) radiations was evaluated. The photodegradation process was monitored by a capillary electrophoresis (CE) method able to provide the enantioresolution of the rac-nicardipine. Enantioresolution was achieved using the mixture 3.0% sulfate-beta-cyclodextrin (SbetaCD) and 2.0% heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (TMbetaCD) as chiral selector in 20mM triethanolammonium phosphate solution (pH 3.0). The photostability studies were carried out on inclusion complexes of rac-nicardipine with alpha-cyclodextrin (alphaCD), beta-cyclodextrin (betaCD), gamma-cyclodextrin (gammaCD), hydroxypropyl-alpha-cyclodextrin (HPalphaCD), hydroxypropyl-beta-cyclodextrin (HPbetaCD), hydroxypropyl-gamma-cyclodextrin (HPgammaCD), (2-hydroxyethyl)-beta-cyclodextrin (HEbetaCD) and methyl-beta-cyclodextrin (MbetaCD). A photoprotective effect was observed by betaCD, HPalphaCD, HEbetaCD, whereas gammaCD, MbetaCD, HPbetaCD and HPgammaCD did not affect the nicardipine photostability. Conversely, alphaCD was found to favour the drug photodegradation. Evidences for CDs-mediated stereoselective photodegradation of rac-nicardipine were observed only for the beta-CD complex. In this case, two distinct photodegradation profiles, with two different kinetic constants (k), were observed for the nicardipine enantiomers.


Subject(s)
Cyclodextrins/analysis , Cyclodextrins/radiation effects , Nicardipine/analysis , Nicardipine/radiation effects , Ultraviolet Rays , Biotransformation/radiation effects , Cyclodextrins/metabolism , Drug Stability , Electrophoresis, Capillary/methods , Nicardipine/metabolism
6.
Electrophoresis ; 22(15): 3243-50, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11589286

ABSTRACT

Capillary electrophoresis (CE) was applied to photostability studies on rac-nicardipine, a dihydropyridine chiral drug. CE methods were developed able to provide the enantioresolution of drug and its separation from the photodegradation products. Enantioresolution was achieved using 5% sulfated-beta-cyclodextrin (S-beta-CD) as chiral selector in 20 mM triethanolammonium phosphate solution (pH 3). The photostability studies were carried out on inclusion complexes of rac-nicardipine with beta-cyclodextrin (beta-CD) and (2-hydroxypropyl)-beta-cyclodextrin (HP-beta-CD) in aqueous solutions (pH 7.4 and 5). The CE analysis of the solutions exposed to UV-A and UV-B radiations showed a photoprotective effect by beta-CD; conversely, HP-beta-CD proved to favor the drug photodegradation. Moreover, evidences for CDs-mediated stereoselective photodegradation of rac-nicardipine were obtained. In fact, two distinct photodegradation profiles were observed for the nicardipine enantiomers in the presence of the CDs. The photodegradation was found to follow an apparent first-order kinetics and two different kinetic constants (k) were obtained for the two enantiomers. After exposure to UV-A and UV-B radiations, the solutions contained residual nicardipine with a significant change in the enantiomeric ratio; this effect was depending on the CD used for the inclusion complexation.


Subject(s)
Cyclodextrins/chemistry , Electrophoresis, Capillary , Nicardipine/chemistry , Photochemistry , beta-Cyclodextrins , 2-Hydroxypropyl-beta-cyclodextrin , Drug Stability , Electrophoresis, Capillary/methods , Hydrogen-Ion Concentration , Kinetics , Solubility , Solutions , Stereoisomerism , Ultraviolet Rays
7.
J Chromatogr A ; 916(1-2): 175-83, 2001 May 04.
Article in English | MEDLINE | ID: mdl-11382289

ABSTRACT

A micellar electrokinetic chromatographic (MEKC) method was developed for the quantification of mesalazine or 5-aminosalicylic acid (5-ASA) and its major impurities 3-aminosalicylic acid, salicylic acid, sulfanilic acid and 4-aminophenol. The optimisation of the experimental conditions was carried out considering some important requirements: resolution, reproducibility, detection limits of 0.1% (m/m) or less, short total analysis time. Preliminary investigations employing sodium dodecyl sulfate (SDS) as surfactant did not lead to the necessary resolution of the studied compounds; the addition of tetrabutylammonium bromide (TBAB) to the SDS micellar system resulted in the complete separation of all the compounds. The effects on the separation by several parameters such as TBAB concentration, SDS concentration, background electrolyte pH and concentration, were evaluated. Using a fused-silica capillary (8.5 cm effective length) complete analysis was obtained in a very short time. Under the optimised final conditions [120 mM 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid buffer, pH 10.20, 65 mM SDS in the presence of 55 mM TBAB and 5% methanol] the method was validated for specificity, precision, linearity, limits of detection and quantitation, and robustness: the 5-ASA related impurities can be quantified at least at the 0.1% (m/m) level.


Subject(s)
Chromatography, Micellar Electrokinetic Capillary/methods , Indicators and Reagents/chemistry , Mesalamine/chemistry , Ions , Reproducibility of Results , Sensitivity and Specificity , Temperature
8.
J Anal Toxicol ; 25(2): 99-105, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11300514

ABSTRACT

Screening methods based on liquid chromatography (HPLC) and capillary electrophoresis (CE) have been developed for the identification and determination of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, and 3,4-methylenedioxyethylamphetamine in illicit tablets. Diethylpropione, (-)-ephedrine, and 3-amino-1-phenylbutane were also included in the study as amphetamino-related compounds. The HPLC-diode-array detection method involved on-line photochemical derivatization to enhance the selectivity of detection allowing amphetamines to be distinguished from related compounds such as diethylpropione (amfepramone). When the CE approach was adopted, two identification parameters (UV spectra and migration index) were used and the enantioresolution of the racemic amphetamines was achieved using hydroxypropyl-beta-cyclodextrin as chiral selector.


Subject(s)
Amphetamine/analysis , Chromatography, High Pressure Liquid , Electrophoresis, Capillary , Substance Abuse Detection/methods , Humans
9.
J Pharm Biomed Anal ; 24(5-6): 863-70, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11248479

ABSTRACT

The analysis of donepezil, a centrally acting acetylcholine esterase inhibitor, is described by a CZE method suitable for applications in pharmaceutical field. A rapid migration of the analyte was obtained under acidic conditions (pH 3.0); with detection wavelength of 320 nm a LOD of 0.8 x 10(-3) mg/ml was provided. Applications on real sample (pharmaceuticals) were carried out using two different instruments with comparable results in terms of reproducibility and accuracy. The use of chiral selectors in the running buffer allowed the enantioseparation of donepezil; charged cyclodextrins (carboxymethyl-beta-cyclodextrin and sulfated-beta-cyclodextrin) were suitable for the chiral resolution of the analyte. Interesting results were also obtained using human serum albumin. The protein-based CE enantioseparation was carried out at pH 7.4 avoiding the partial filling technique due to the good absorptivity of donepezil at 320 nm. Interestingly, the use of bicine as BGE provided a significative improvement in the enantioresolution compared to that obtained by phosphate buffer.


Subject(s)
Cholinesterase Inhibitors/analysis , Electrophoresis, Capillary/methods , Indans/analysis , Piperidines/analysis , Cholinesterase Inhibitors/chemistry , Cholinesterase Inhibitors/isolation & purification , Donepezil , Indans/chemistry , Indans/isolation & purification , Piperidines/chemistry , Piperidines/isolation & purification , Reproducibility of Results , Sensitivity and Specificity , Stereoisomerism
10.
J Inherit Metab Dis ; 23(2): 120-8, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10801053

ABSTRACT

Mutation analysis was performed on DNA from 31 Turkish children with profound biotinidase deficiency who were symptomatic or ascertained by newborn screening. The 98G:del7ins3 mutation is common in clinically ascertained children in both the United States and Turkish populations, but a unique common mutation, R79C, is found only in the Turkish children identified both clinically and by newborn screening. Another frequently occurring mutation, T532M, is only observed in the Turkish newborn screening group. There are four other less frequent novel mutations identified in the Turkish population. Interestingly, the Q456H and the A171T:D444H double mutation, which are the most common mutations found in the US newborn screening population and have not been observed in symptomatic children, do occur in clinically ascertained children in the Turkish population, although the double mutation may be associated with milder and/or later-onset symptoms.


Subject(s)
Amidohydrolases/deficiency , Biotin/metabolism , Metabolism, Inborn Errors/genetics , Mutation/genetics , Biotinidase , Child , Consanguinity , DNA/analysis , DNA/genetics , DNA Mutational Analysis , Humans , Infant, Newborn , Neonatal Screening , Turkey
12.
Hum Hered ; 50(2): 102-11, 2000.
Article in English | MEDLINE | ID: mdl-10799968

ABSTRACT

Biotinidase deficiency is an autosomal recessive defect in the recycling of biotin that can lead to a variety of neurologic and cutaneous symptoms. The disease can be prevented or effectively treated with exogenous biotin. The biotinidase locus (BTD) has been maped to 3p25 by in situ hybridization. The gene has been cloned, the coding region sequenced, the genomic organization determined, and a spectrum of mutations has been characterized in more than 90 individuals with profound or partial biotinidase deficiency. We have conducted haplotype analysis of 10 consanguineous and 39 nonconsanguineous probands from the United States and 8 consanguineous probands from Turkey to localize BTD with respect to polymorphic markers on 3p and to investigate the origins of five common mutations. The inbred probands were homozygous for overlapping regions of 3p ranging in size from 1.1 to 80 cM which were flanked most narrowly by D3S1259 and D3S1293. Radiation hybrids and haplotype analysis of markers within this region suggest that BTD is located within a 0.1-cM region flanked by D3S3510 and D3S1286. The radiation hybrid data suggest that the BTD gene is oriented 5' to 3' between the centromere and the 3p telomere. Association studies indicate that the gene is closer to a third locus D3S3613 than D3S3510, two markers which cannot be resolved by existing linkage data. The BTD locus and D3S3613 must therefore lie between D3S3510 and D3S1286. Comparison of haplotypes reveals evidence for possible founder effects for four of the five common mutations.


Subject(s)
Amidohydrolases/genetics , Chromosome Mapping , Mutation , Adult , Amidohydrolases/deficiency , Biotinidase , Consanguinity , Female , Genotype , Haplotypes , Humans , Infant, Newborn , Male
13.
Pediatr Res ; 46(1): 20-7, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10400129

ABSTRACT

Biotinidase deficiency is an autosomal recessive disorder of biotin metabolism that can lead to varying degrees of neurologic and cutaneous symptoms when untreated. Because this disorder meets the criteria for newborn screening, many states and countries perform this testing. Because newborn screening should result in complete ascertainment of mutations causing profound biotinidase deficiency (less than 10% of mean normal serum activity), we compared the mutations in a group of 59 children with profound biotinidase deficiency who were identified by newborn screening in the United States with 33 children ascertained by exhibiting symptoms. Of the 40 total mutations identified among the two populations, four mutations comprise 59% of the disease alleles studied. Two of these mutations occur in both populations, but in the symptomatic group at a significantly greater frequency. The other two common mutations occur only in the newborn screening group. Because two common mutations do not occur in the symptomatic population, it is possible that individuals with these mutations either develop mild or no symptoms if left untreated. However, inasmuch as biotin treatment is inexpensive and innocuous, it is still recommended that all children with profound biotinidase deficiency be treated.


Subject(s)
Amidohydrolases/deficiency , Amidohydrolases/genetics , Genetic Testing , Multiple Carboxylase Deficiency/diagnosis , Mutation , Neonatal Screening , Amino Acid Substitution , Biotinidase , Frameshift Mutation , Humans , Infant, Newborn , Multiple Carboxylase Deficiency/epidemiology , Multiple Carboxylase Deficiency/genetics , Point Mutation , Polymerase Chain Reaction , United States/epidemiology
14.
J Chromatogr A ; 844(1-2): 361-9, 1999 Jun 04.
Article in English | MEDLINE | ID: mdl-10636700

ABSTRACT

The ability of capillary zone electrophoresis in the development of analytical methods devoted to the quality control of the thiol drug penicillamine is shown. Using 50 mM phosphate running buffer (pH 2.5), good quantitations of underivatized penicillamine and its disulfide were achieved; detection at 200 nm allowed checking the presence of the disulfide impurity in pharmaceuticals. The use of 1,1-[ethenylidenebis(sulfonyl)]bis-benzene as a thiol specific reagent resulted in an increased sensitivity for the quantitation of D-penicillamine (limit of detection at 200 nm wavelength was 1.5 microM). Introducing beta-cyclodextrin as chiral selector in the running buffer, enantioseparation of D-L-penicillamine was obtained; for this purpose (+)-camphor-10-sulfonic acid, a chiral ion-pairing reagent, was found to be an essential additive in obtaining a baseline separation. The resulting enantioseparative system was validated in order to evaluate the presence of the toxic L-penicillamine enantiomer in pharmaceutical samples.


Subject(s)
Electrophoresis, Capillary/methods , Penicillamine/analysis , Technology, Pharmaceutical , Disulfides/analysis , Indicators and Reagents , Penicillamine/chemistry , Quality Control , Sensitivity and Specificity , Stereoisomerism , Sulfhydryl Compounds
15.
Mol Genet Metab ; 64(2): 152-4, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9705240

ABSTRACT

The only known Japanese child with biotinidase deficiency was identified by newborn screening in Japan. He has 10.8% of mean normal serum biotinyl-hydrolase activity and trace biotinyl-transferase activity. The mutation results in 16% of normal cross-reacting material in serum with antibody to purified normal biotinidase. He is homozygous for a unique mutation, A1466 > C (Asn489Thr) in exon 4 of the biotinidase gene. The mutation appears to abolish a putative glycosylation site in a region in which other missense mutations have been identified, indicating that this region of the enzyme must be important for enzyme activity. This mutation may affect secretion or stability of the enzyme in serum. Interestingly, this child is now 8 years old, has not been on biotin supplementation for 3 years, and has remained asymptomatic.


Subject(s)
Amidohydrolases/deficiency , Amidohydrolases/genetics , Amidohydrolases/blood , Amino Acid Substitution , Asparagine/genetics , Binding Sites/genetics , Biotinidase , Child , DNA Mutational Analysis , Glycosylation , Homozygote , Humans , Japan , Male , Mutation/genetics , Threonine/genetics
16.
Mamm Genome ; 9(4): 327-30, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9530634

ABSTRACT

Biotinidase cleaves biotin from biocytin, thereby recycling the vitamin. We have determined the structure of the human biotinidase gene. A genomic clone, containing three exons that code for the mature enzyme, was obtained by screening a human genomic bacteriophage library with the biotinidase cDNA by plaque hybridization. To obtain a clone containing the most 5' exon of the biotinidase cDNA, a human PAC library by PCR was screened. The human biotinidase gene is organized into four exons and spans at least 23 kb. The 5'-flanking region of exon 1 contains a CCAAT element, three initiator sequences, an octamer sequence, three methylation consensus sites, two GC boxes, and one HNF-5 site, but has no TATA element. The region from nt -600 to +400 has features of a CpG island and resembles a housekeeping gene promoter. The structure and sequence of this gene are useful for identifying and characterizing mutations that cause biotinidase deficiency.


Subject(s)
Amidohydrolases/genetics , Base Sequence , Biotinidase , DNA, Complementary , Exons , Humans , Introns , Molecular Sequence Data , Polymerase Chain Reaction
17.
Prenat Diagn ; 18(2): 117-22, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9516011

ABSTRACT

Biotinidase deficiency is characterized by neurological and cutaneous abnormalities that can be prevented or ameliorated by oral biotin therapy. A child with biotinidase deficiency went undiagnosed for a long period and has irreversible neurological deficits despite biotin treatment. This child is homozygous for the most common mutation (G98:d7i3) found in symptomatic children with the disorder. The parents insisted on having prenatal diagnosis in a subsequent pregnancy to alleviate their anxiety about having another affected child. Mutation analysis of DNA obtained directly from amniotic fluid and from cultured amniocytes revealed that the fetus was heterozygous for the mutation. Maternal cell contamination of the amniocytes was excluded by genotype analysis. Biotinidase activity in extracts of cultured amniocytes revealed 40 per cent of mean normal activity. At birth, the infant was confirmed to be heterozygous by serum enzyme analysis. This is the first report of the use of molecular analysis for the prenatal diagnosis for biotinidase deficiency.


Subject(s)
Amidohydrolases/deficiency , Genetic Carrier Screening , Metabolism, Inborn Errors/diagnosis , Prenatal Diagnosis , Amidohydrolases/blood , Amidohydrolases/genetics , Amniotic Fluid/chemistry , Amniotic Fluid/cytology , Biotinidase , Cells, Cultured , DNA Mutational Analysis , Female , Genotype , Homozygote , Humans , Infant , Male , Mutation , Pregnancy
18.
J Pediatr ; 132(2): 362-5, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9506660

ABSTRACT

Children with biotinidase deficiency usually exhibit symptoms at several months to years of age. We describe four children who had symptoms later in childhood or during adolescence; they had motor limb weakness, spastic paresis, and eye problems, such as loss of visual acuity and scotomata, rather than the more characteristic symptoms observed in young untreated children with the disorder. These older children each have different mutations, but they are the same as those of children who have exhibited symptoms at an early age. Biotinidase deficiency should be considered in older children who suddenly experience limb weakness and/or spastic paresis and eye symptoms.


Subject(s)
Acyltransferases/deficiency , Amidohydrolases/deficiency , Metabolism, Inborn Errors , Acyltransferases/genetics , Adolescent , Age of Onset , Amidohydrolases/genetics , Biotinidase , Child , Female , Humans , Male , Metabolism, Inborn Errors/genetics , Mutation
19.
Hum Mutat ; 11(5): 410, 1998.
Article in English | MEDLINE | ID: mdl-10206677

ABSTRACT

Biotinidase deficiency is inherited as an antosomal recessive trait that, unless treated with pharmacologic doses of biotin, can result in neurologic and cutaneous symptoms. We have identified two new mutations in exon D of the biotinidase gene of children with profound biotinidase deficiency ascertained by newborn screening. Transition 511G->A near the 5' end of exon D results in a substitution of threonine for alanine 171 (A171T) and transversion 1330G->C occurs close to the 3' end of exon D causing a substitution of histidine for aspartic acid 444 (D444H). The D444H mutation was detected in four individuals from our normal population whose mean serum biotinidase activity is 5.25 nmol/min/ml, which is significantly lower than the mean normal activity (7.1 nmol/min/ml). We calculated that this mutation causes a 52% loss of activity in the aberrant enzyme. Twenty-three individuals with the D444H mutation were found by allele specific oligonucleotide analysis of DNA from 296 randomly-selected, anonymous dried-blood spots. We estimate the frequency of this allele in the general population to be 0.039. In contrast, no individuals in 376 have the A171T mutation. Fourteen children (eleven probands and three siblings) out of the 31 enzyme-deficient children have both the A171T and D444H mutations. Both mutations are inherited from a single parent as a double mutation allele. The nine families in which this allele was identified are of mostly European ancestry, although the mutation cannot be attributed to a specific nationality or ethnic group. The serum of a child who is homozygous for the double mutation allele has very little CRM and the aberrant enzyme has very low biotinylhydrolase activity and no botinyl-transferase activity. This double mutation allele (A171T and D444H) is a common cause of profound biotinidase deficience in children ascertained by newborn screening in the United States.


Subject(s)
Acyltransferases/deficiency , Mutation/genetics , Neonatal Screening , Acyltransferases/genetics , Alanine/genetics , Aspartic Acid/genetics , Histidine/genetics , Humans , Infant, Newborn , Threonine/genetics
20.
Pediatr Res ; 42(6): 840-8, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9396567

ABSTRACT

Biotinidase deficiency is an autosomal recessively inherited disorder that results in the inability to recycle the vitamin biotin. The disorder can cause neurologic and cutaneous abnormalities that can be treated effectively with pharmacologic doses of biotin. We identified 21 mutations that cause profound biotinidase deficiency in 37 symptomatic children (30 different probands and 7 siblings), as well as provide relevant biochemical and clinical information for each child. The two most common mutations (G98:d7i3 and R538C) were found in 31 of 60 alleles (52%), whereas the remainder of the alleles are accounted for by the 19 other unique mutations. Serum samples were available from 18 children, of these 11 had no detectable cross-reacting material (CRM) to antibody prepared against normal human serum biotinidase, three had reduced quantities of CRM and four had normal quantities of CRM in serum. All of these mutations result in complete absence of biotinyl-transferase activity in serum. Two polymorphisms were also identified in normal individuals. It is apparent that a child who inherits any of these mutations, either in the homozygous state or in combination, can develop the clinical features of the disorder if untreated. There are, however, no clear genotype/phenotype correlations that would allow for the prediction of the type, severity, or age of onset of symptoms.


Subject(s)
Acyltransferases/genetics , Amidohydrolases/genetics , Acyltransferases/blood , Acyltransferases/deficiency , Amidohydrolases/blood , Amidohydrolases/deficiency , Biotinidase , Child , Genetic Testing , Genotype , Humans , Mutation , Phenotype , Sequence Analysis, DNA
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