The cystic fibrosis transmembrane conductance regulator (CFTR) gene exhibits a tightly regulated pattern of expression in human epithelial cells. The mechanism of this regulation is complex and is likely to involve a number of genetic elements that effect temporal and spatial expression. To date none of the elements that have been identified in the CFTR promoter regulate tissue-specific expression. We have identified a putative regulatory element within the first intron of the CFTR gene at 181+10kb. The region containing this element was first identified as a DNase I hypersensitive site that was present in cells that express the CFTR gene but absent from cells not transcribing CFTR. In vitro analysis of binding of proteins to this region of DNA sequence by gel mobility shift assays and DNase I footprinting revealed that some proteins that are only present in CFTR-expressing cells bound to specific elements, and other proteins that bound to adjacent elements were present in all epithelial cells irrespective of their CFTR expression status. When assayed in transient expression systems in a cell line expressing CFTR endogenously, this DNA sequence augmented reporter gene expression through activation of the CFTR promoter but had no effect in nonexpressing cells.

Human MutationVolume 6, Issue 2 p. 170-176 Mutation in Brief Identification of seven novel mutations associated with metachromatic leukodystrophy Maria Luiza Barth, Maria Luiza Barth Division of Medical and Molecular Genetics, UMDS-Guy's Campus, London SEl 9RT, UKSearch for more papers by this authorAnthony Fensom, Anthony Fensom Division of Medical and Molecular Genetics, UMDS-Guy's Campus, London SEl 9RT, UKSearch for more papers by this authorAnn Harris, Corresponding Author Ann Harris Paediatric Molecular Genetics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK; Fax: 44-865-222-626Paediatric Molecular Genetics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK; Fax: 44-865-222-626Search for more papers by this author Maria Luiza Barth, Maria Luiza Barth Division of Medical and Molecular Genetics, UMDS-Guy's Campus, London SEl 9RT, UKSearch for more papers by this authorAnthony Fensom, Anthony Fensom Division of Medical and Molecular Genetics, UMDS-Guy's Campus, London SEl 9RT, UKSearch for more papers by this authorAnn Harris, Corresponding Author Ann Harris Paediatric Molecular Genetics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK; Fax: 44-865-222-626Paediatric Molecular Genetics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK; Fax: 44-865-222-626Search for more papers by this author First published: 1995 https://doi.org/10.1002/humu.1380060210Citations: 22AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume6, Issue21995Pages 170-176 RelatedInformation

Arylsulphatase A (ASA, EC3.1.6.1) is a lysosomal enzyme that catalyses cerebroside sulphate degradation. ASA deficiency is associated with metachromatic leucodystrophy (MLD), a rare autosomal recessive disorder, which is characterised by the storage of cerebroside sulphate. Low ASA activities can be also observed in clinically healthy persons, a condition termed ASA pseudodeficiency. Two mutations responsible for the majority of pseudodeficiency alleles have been defined in the ASA gene. These are both A --> G transitions. One causes an asparagine to serine substitution (N350S). The second changes the first polyadenylation signal downstream of the stop codon (1524 + 95 A --> G), which causes a severe deficiency of one ASA mRNA species. The incidence of the pseudodeficiency allele is estimated to be high in the general population and can be found in families carrying MLD associated mutations. We report a reliable stratagem for detecting the two PD associated mutations separately, which we have applied to a healthy population. Two homozygotes for the N350S and 1524 + 95 A --> G mutations were detected, which gives a population frequency of 2.6%. The overall frequencies of the ASA-PD mutations were shown to be 17.5% for the N350S change and 13.0% for the 1524 + 95 A --> G change, estimating each mutation separately. In addition, the frequency of both PD associated mutations occurring together on the same chromosome was found to be 12.3% in our population. The study has also allowed us to establish a new control ASA activity range, which was based on assay of blood from persons who had been shown at the DNA level not to carry ASA PD associated mutations.

Metachromatic leucodystrophy is an autosomal recessive degenerative disease of the nervous system caused by the deficiency of the lysosomal enzyme arylsulphatase A (ARSA). We report here on the high incidence of late infantile MLD among Muslim Arabs originating from Jerusalem, most probably because of a founder effect. All the patients were found to be homozygous for 459 + 1 G-->A, a mutation which destroys the splice donor site of exon 2 of the ARSA gene. This mutation has been reported to be the most common mutation causing MLD. We studied the ARSA haplotype defined by three intragenic polymorphic sites in DNA samples from Muslim Arab patients from Jerusalem, a Christian Arab patient originating from the region, and eight other white patients, all homozygous for the 459 + 1 G-->A mutation. All the alleles carried the same haplotype which is in complete linkage disequilibrium with the mutation. This finding indicates a common origin for the 459 + 1 G-->A mutation which may have been introduced into Jerusalem at the time of the Crusades.

The lysosomal storage disorders share one common characteristic: the accumulation of a par-ticular substrate inside the lysosome. Each disorder in this group is caused either by the deficiency of a lysosomal enzyme responsible for one step of the degradation pathway of a substrate, lack of a transporter involved in the movement of a small molecule across the lysosomal membrane, or absence of a low molecular weight activator protein required for in vivo hydrolysis of a sphingolipid. There are now more than 30 lysosomal storage diseases where the biochemical defect is well known and fully characterised. In this group, metachromatic leucodystrophy (MLD) is an autosomal recessive disorder that has been ex-tensively studied over the past few decades.' The overall incidence of MLD is estimated to be 1:40 0002 and the enzymatic defect associated with this disease was discovered in the 1960s. More recently, the molecular biology of MLD has been elucidated, adding to the well established biochemical knowledge. Systematic mutation analyses have been carried out in an attempt to establish genotype-phenotype relationships. A clear understanding ofthe molecular basis of this disease will be helpful in the genetic counselling of affected families.

Novel predicted disease-causing mutations have been defined in three patients with metachromatic leukodystrophy (MLD). The first new mutation is a C-->A change at base 884 in exon 5 of the arylsulphatase A (ASA) gene causing a serine to tyrosine substitution at position 295 of the protein (S295Y). A late-infantile MLD patient was found to be homozygous for this mutation. The second mutation is a G-->A substitution at nucleotide 1144 in exon 7, that causes a glutamic acid to lysine substitution at amino acid 382 (E382K). A juvenile MLD patients was found to be homozygous for this mutation. Finally an adult MLD patient has been shown to be heterozygous for two novel point mutations in exon 3. These are both C-->T changes at position 635 and 671 that result in alanine to valine substitutions at amino acids 212 (A212V) and 224 (A224V) of the ASA protein.

Twenty-six patients with Gaucher's disease diagnosed in the United Kingdom and two obligate carriers, all of non-Jewish origin, were screened for the two common disease causing mutations and two rarer mutations in the glucocerebrosidase gene. These mutations are referred to as N370S, L444P, Ins84G, and 1066 + 1G-->A, respectively. The results showed that out of 54 alleles screened, 26% were N370S, 35% were L444P, and the remaining 39% were rare or undefined. The results also showed a clear correlation between the presence of at least one N370S allele and mild disease.

Journal of Inherited Metabolic DiseaseVolume 14, Issue 3 p. 400-402 Short Communication Seven-year experience of a reference laboratory for detection of inborn errors of metabolism in Brazil R. Giugliani, R. Giugliani Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorJ. C. Dutra, J. C. Dutra Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorM. L. Barth, M. L. Barth Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorC. S. Dutra-Filho, C. S. Dutra-Filho Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorS. L. Goldenfum, S. L. Goldenfum Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorM. Wajner, M. Wajner Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this author R. Giugliani, R. Giugliani Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorJ. C. Dutra, J. C. Dutra Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorM. L. Barth, M. L. Barth Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorC. S. Dutra-Filho, C. S. Dutra-Filho Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorS. L. Goldenfum, S. L. Goldenfum Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this authorM. Wajner, M. Wajner Medical Genetic Unit, Clinical Hospital of Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre RS, 90210 BrazilSearch for more papers by this author First published: 01 May 1991 https://doi.org/10.1007/BF01811714Citations: 3AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References 1Giugliani R., Dutra-Filho C. S., Barth M. L., Dutra J. C., Wajner M., Wannmacher C. M. D., Montagner L. T. (1989) Inborn errors of metabolism: sensitivity of screening tests in high risk patients. Clin. Pediatr., 28: 494–497. 2Levy H. L., Shih V. E., Madigan P. M. (1972) Massachusetts metabolic disorders screening program. I. Techniques and results of urine screening. Pediatrics, 49: 825–836. Citing Literature Volume14, Issue3May 1991Pages 400-402 ReferencesRelatedInformation

Very low serum levels of high density lipoprotein cholesterol ranging from 8.6 to 13.9 mg/dl were detected in four out of 12 sibs of a Brazilian kindred with the non-neuropathic form of Niemann-Pick disease. Hepatosplenomegaly, interstitial infiltration of the lungs, absence of neurological signs, sea-blue histiocytes in the bone marrow and liver, and high values for serum acid phosphatase (18 to 32 U/l) were common to all affected children. Leucocyte acid sphingomyelinase activity ranged from 3.6 to 6.5% of mean control values, and fibroblast activity from 9 to 13% of mean controls. The parents had low-normal levels. The relationship between these findings is unclear and deserves further investigation.

Lysosomal storage diseases (LSD) are a group of more than 40 disorders, many of them with overlapping phenotype, in which clinical diagnosis is often difficult. Definitive diagnosis is based on enzyme assays, a large number of such assays usually being necessary during the investigation of each patient. In addition, there will frequently be a need for tissue culture in order to provide enough material for analysis. Taking into account these difficulties, we designed a flowchart for the detection of LSD that is based on 2 sets of tests requiring only random urine and heparinized blood. Here we describe this routine and report the results of its application to 105 Brazilian patients in whom a LSD was suspected. We think that the application of this rationale represents a saving of work and costs, and should be of special interest to genetic centers in developing countries.

We estimated the sensitivity of a screening procedure (SP) for inborn errors of metabolism (IEM) in 566 referred, high-risk patients. The 143 (25.3% of the total sample) patients with initial abnormal results in at least one screening test (ST) were recalled for further investigations. An IEM was diagnosed in 40.6 percent of the 106 patients who came for reevaluation. In 114 of the remaining 423 patients who had normal initial ST, an IEM was still suspected on basis of clinical, radiological, and/or laboratory findings and was confirmed in 30 of such patients (5.3% of the total sample and 7.1% of the patients with normal results in the SP). The sensitivity of the SP was estimated maximally as 67.4 percent and the efficiency as 80.4 percent. Twenty-five of the 30 cases undetected with the SP were patients with sphingolipidoses. The simple inclusion of thin-layer chromatography of urinary oligosaccharides in the SP should allow the detection of at least one half of these cases, increasing its sensitivity by 14.1 percent and its efficiency by 4.6 percent. In at least 7.1 percent of patients with an initial normal ST, an IEM was detected. These would have remained undiagnosed if the limitations of the SP employed had not been fully understood.

We estimated the sensitivity of a screening procedure (SP) for inborn errors of metabolism (IEM) in 566 referred, high-risk patients. The 143 (25.3% of the total sample) patients with initial abnormal results in at least one screening test (ST) were recalled for further investigations. An IEM was diagnosed in 40.6 percent of the 106 patients who came for reevaluation. In 114 of the remaining 423 patients who had normal initial ST, an IEM was still suspected on basis of clinical, radiological, and/or laboratory findings and was confirmed in 30 of such patients (5.3% of the total sample and 7.1% of the patients with normal results in the SP). The sensitivity of the SP was estimated maximally as 67.4 percent and the efficiency as 80.4 percent. Twenty-five of the 30 cases undetected with the SP were patients with sphingolipidoses. The simple inclusion of thin-layer chromatography of urinary oligosaccharides in the SP should allow the detection of at least one half of these cases, increasing its sensitivity by 14.1 percent and its efficiency by 4.6 percent. In at least 7.1 percent of patients with an initial normal ST, an IEM was detected. These would have remained undiagnosed if the limitations of the SP employed had not been fully understood.