Device and method for nonlinear ultrasonic measurements on highly irradiated 304 stainless steel specimens in a hot cell environment.

This paper reports on the first successful nonlinear ultrasonic measurement on highly irradiated specimens in a hot cell environment. The specimens are ANSI 304 stainless steel specimens for which the microstructure characterization and ultrasonic velocity measurement have been previously conducted. The critical part of this research is the development of an automatic fixture device that can facilitate repeatable loading and unloading to place the contact ultrasonic transducers on and off of the specimen. The key step to achieve high measurement repeatability is a careful adjustment of the support-spring constants such that the contact force at the interface between the transducer face and specimen surface is as uniform and constant as possible. The longitudinal ultrasonic velocities, which are obtained as a by-product of the nonlinear ultrasonic measurements, show a level of random variation in terms of (max-min)/average (%) below 0.2%, and the velocity distributions and magnitudes are in good agreement with those from the previous work. The ultrasonic nonlinearity parameters show the level of random variation below 4.7%, which is extremely low, considering that the measurements are conducted in a hot cell environment. The nonlinearity parameters also show a strong dependence on the measurement location in a particular specimen with respect to the radiation source, demonstrating a possible inhomogeneous microstructure evolution in these 12.7 mm thick specimens. This research demonstrates the feasibility of making nonlinear ultrasonic measurement on highly radioactive materials and/or in a highly radioactive environment using the device and procedure developed.

Pearson syndrome and the role of deletion dimers and duplications in the mtDNA.

Pearson syndrome is an often fatal multisystem disease associated with mitochondrial DNA rearrangements. Here we report a patient with a novel mtDNA deletion of 3.4 kb ranging from nucleotides 6097 to 9541 in combination with deletion dimers. The mutation percentage in different tissues (blood, muscle and liver) varied between 64% and 95%. After a remission period of about a year, the patient suddenly died at the age of 3 years owing to a severe lactic acidosis. A second patient with a previously reported deletion of 8 kb and a milder phenotype was found to have mitochondrial duplications and died at the age of 10 years. From these data and data from previous reports, we hypothesize that duplications might be beneficial in the clinical course of the disease and in life expectancy.

Multiplex PCR of polymorphic markers flanking the CFTR gene; a general approach for preimplantation genetic diagnosis of cystic fibrosis.

Cystic fibrosis (CF) is the first monogenic disorder for which single cell preimplantation genetic diagnosis (PGD) has been successfully applied. The spectrum of mutations in CF is extremely heterogeneous, and hence, the development of mutation-specific PGD protocols is impracticable. The current study reports the development and evaluation of a general multiplex marker polymerase chain reaction (PCR) protocol for PGD of CF. Four closely linked highly polymorphic (CA)(n) repeat markers D7S523, D7S486, D7S480 and D7S490, flanking the cystic fibrosis transmembrane regulator (CFTR) gene, were used. In 99% of the single cells tested (100 leukocytes and 50 blastomeres), multiplex PCR results were obtained and the overall allelic drop out (ADO) rate varied from 2 to 5%. After validation for the presence of ADO and additional alleles, 95% of the multiplex PCR results were accepted to construct the marker genotypes. Depending on the genotype of the couple, and taking into account the embryos lost for transfer due to validation criteria (5%), ADO (0-2%) and single recombination (1.1-3%), in general >90% of the embryos could be reliably genotyped by PGD using a single blastomere. The risk of misdiagnosis equals the chance of a double recombination between informative flanking markers and is <0.05%. Therefore, this polymorphic and multi-allelic marker system is a reliable and generally applicable alternative for mutation-directed PGD protocols. Furthermore, it provides a test for the origin of the detected genotype and also gives an indication of the chromosomal ploidy status of the blastomere tested.