van der Loop FT, Heidet L, Timmer ED, van den Bosch BJ, Leinonen A, Antignac C, Jefferson JA, Maxwell AP, Monnens LA, Schroder CH, Smeets HJ. Autosomal dominant Alport syndrome caused by a COL4A3 splice site mutation. Kidney Int. 2000 Nov;58(5):1870-5.

van Greevenbroek MM, van der Kallen CJ, Geurts JM, Janssen RG, Buurman WA, de Bruin TW. Soluble receptors for tumor necrosis factor-alpha (TNF-R p55 and TNF-R p75) in familial combined hyperlipidemia. Atherosclerosis. 2000 Nov;153(1):1-8.

van der Kallen CJ, Cantor RM, van Greevenbroek MM, Geurts JM, Bouwman FG, Aouizerat BE, Allayee H, Buurman WA, Lusis AJ, Rotter JI, de Bruin TW. Genome scan for adiposity in Dutch dyslipidemic families reveals novel quantitative trait loci for leptin, body mass index and soluble tumor necrosis factor receptor superfamily 1A. Int J Obes Relat Metab Disord. 2000 Nov;24(11):1381-91.


van Den Bosch BJ, de Coo RF, Scholte HR, Nijland JG, van Den Bogaard R, de Visser M, de Die-Smulders CE, Smeets HJ. Mutation analysis of the entire mitochondrial genome using denaturing high performance liquid chromatography . Nucleic Acids Res. 2000 Oct 15;28(20):E89.


Geurts JM, Janssen RG, van Greevenbroek MM, van der Kallen CJ, Cantor RM, Bu X, Aouizerat BE, Allayee H, Rotter JI, de Bruin TW. Identification of TNFRSF1B as a novel modifier gene in familial combined hyperlipidemia. Hum Mol Genet. 2000 Sep 1;9(14):2067-74.

Procaccio V, Lescuyer P, Bourges I, Beugnot R, Duborjal H, Depetris D, Mousson B, Montfort MF, Smeets H, De Coo R, Issartel JP. Human NDUFS3 gene coding for the 30-kDa subunit of mitochondrial complex I: genomic organization and expression. Mamm Genome. 2000 Sep;11(9):808-10.

Vermeire S, Peeters M, Vlietinck R, Parkes M, Satsangi J, Jewell D, Rutgeerts P. Exclusion of linkage of Crohn's disease to previously reported regions on chromosomes 12, 7, and 3 in the Belgian population indicates genetic heterogeneity. Inflamm Bowel Dis. 2000 Aug;6(3):165-70. 

Thomis MA, Vlietinck RF, Maes HH, Blimkie CJ, van Leemputte M, Claessens AL, Marchal G, Beunen GP. Predictive power of individual genetic and environmental factor scores. Twin Res. 2000 Jun;3(2):99-108.

Engelen JJ, Marcelis C, Herbergs J, Weber J, Alofs M, Albrechts JC, Hamers AJ. Mosaic telomeric (2;14) association in a child with motor delay. Am J Med Genet. 2000 Jun 19;92(5):318-21.

van den Wijngaard A, Mulder WR, Dijkema R, Boersma CJ, Mosselman S, van Zoelen EJ, Olijve W. Antiestrogens specifically up-regulate bone morphogenetic protein-4 promoter activity in human osteoblastic cells. Mol Endocrinol. 2000 May;14(5):623-33

Dreesen JC, Jacobs LJ, Bras M, Herbergs J, Dumoulin JC, Geraedts JP, Evers JL, Smeets HJ. Multiplex PCR of polymorphic markers flanking the CFTR gene; a general approach for preimplantation genetic diagnosis of cystic fibrosis. Mol Hum Reprod. 2000 May;6(5):391-6.

van den Wijngaard A, Mulder WR, Dijkema R, Boersma CJ, Mosselman S, van Zoelen EJ, Olijve W. Antiestrogens specifically up-regulate bone morphogenetic protein-4 promoter activity in human osteoblastic cells. Mol Endocrinol. 2000 May;14(5):623-33

Jais JP, Knebelmann B, Giatras I, De Marchi M, Rizzoni G, Renieri A, Weber M, Gross O, Netzer KO, Flinter F, Pirson Y, Verellen C, Wieslander J, Persson U, Tryggvason K, Martin P, Hertz JM, Schroder C, Sanak M, Krejcova S, Carvalho MF, Saus J, Antignac C, Smeets H, Gubler MC. X-linked Alport syndrome: natural history in 195 families and genotype-phenotype correlations in males. J Am Soc Nephrol. 2000 Apr;11(4):649-57. 

Beunen G, Thomis M, Maes HH, Loos R, Malina RM, Claessens AL, Vlietinck R. Genetic variance of adolescent growth in stature. Ann Hum Biol. 2000 Mar-Apr;27(2):173-86.

POSTERS

ASHG

Mutation analysis of the entire mitochondrial DNA using Denaturing High Performance Liquid Chromatography (DHPLC).
H. Smeets1, B. Van den Bosch1, J. Nijland1, H. Scholte2, C. DeDie1, R. Van den Bogaard3, M. De Visser3, I. De Coo2. 1) Dept Molecular, Cell Biol & Gen, Univ Maastricht, Maastricht, Netherlands; 2) Depts Child Neurology & Biochemistry, Erasmus Univ, Rotterdam, Netherlands; 3) Depts Clin Genet & Neurol, AMC, Amsterdam, Netherlands. In patients with mitochondrial disease a continuously increasing number of mtDNA mutations and polymorphisms have been identified. Most pathogenic mtDNA mutations are heteroplasmic, resulting in heteroduplexes after PCR amplification of mtDNA. To detect these heteroduplexes, we used the technique of Denaturing High Performance Liquid Chromatography (DHPLC). The entire mtDNA was amplified in 13 fragments of 1-2 kb, digested in fragments of 90-600 bp and resolved at their optimal melting temperature. The sensitivity of the DHPLC system was high with a lowest detectable percentage of 0.5% for the A8344G mutation. The entire muscle mtDNA from 6 patients with mitochondrial disease was screened and 3 mutations were identified. The first patient with a limb-girdle-type myopathy carried an A3302G substitution in the tRNA-Leu(UUR) gene (70% heteroplasmy), the second patient with mitochondrial myopathy and cardiomyopathy a T3271C mutation in the tRNA-Leu(UUR) gene (80% heteroplasmy) and the third patient with Leigh syndrome a T9176C mutation in the ATPase6 gene (93% heteroplasmy). We conclude that DHPLC analysis is a sensitive and specific method to detect heteroplasmic mtDNA mutations. The entire automatic procedure can be completed within 2 days and can also be applied to exclude mtDNA involvement, providing a basis for subsequent investigation of nuclear genes. Subsequently, we optimised the procedure at the single cell level to test for heteroplasmy in single oocytes and embryos to determine the natural mutation load of mtDNA and the possible presence of paternal mtDNA.