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Girdle Mature ((NEW))

We describe a kindred with a rare autosomal dominant myopathy limited to the limb-girdle muscles, beginning insidiously any time from the late second through the sixth decades and followed by slow progression. Pelvifemoral precedes scapulohumeral weakness, and proximal appendicular involvement antedates limited distal paresis. Expressivity varies and includes an asymptomatic myopathy (preclinical or subclinical) and a nonmanifesting carrier state that extends well into the eighth decade. A variety of nonspecific changes are present in muscle on light, enzyme histochemical, and electron microscopic examination; of these changes, "rimmed" or autophagic vacuoles are the most characteristic. We identified one very similar previously reported genealogy. The similarities between the two unrelated families clearly establish this dystrophic process as a distinct genetic entity; their differences suggest genetic heterogeneity.

girdle mature


The girdle of Achnanthes longipes has been studied using scanning and transmission electron microscopy. An examination of 132 isolated bands showed that they can be classified into 3 main morphological types, all of which are usually open at one end, and bear poroids with occlusions recalling those of the valve poroids. The most advalvar band (band 1 or valvocopula) has a single row of elongated poroids on the pars exterior. The poroids have a groove-like appearance in internal views. The band interlocks with the valve following the complementary shape of its pars interior and the internal valve margin. Type 2 bands, which follow the valvocopula in the abvalvar direction, have a ligula or ligula-like expansion at the closed end, with no antiligula; their pars exterior usually bears 2 rows of circular poroids. A type 2 band seems to interlock with the valvocopula via its crenulate pars interior. The crenations may interact internally with the poroids of the valvocopula, or the cavities marking the limit between pars interior and pars exterior of the valvocopula. Presumably, type 2 bands also interlock with other type 2 bands. Type 3 bands follow type 2 bands in the abvalvar direction. They differ from type 2 bands especially in the smooth pars interior, which probably makes the interlock with type 2 bands looser than between type 2 and 1 bands, or between band 1 and the valve. Mature cingula have a total of 6 bands each. In addition to the single type 1 band, there are probably four type 2 bands and one type 3 band.

Butternut (Juglans cinerea L.), or white walnut, has suffered large population declines in the past half-century due to poor regeneration and mortality caused by an introduced fungus, Ophiognomonia clavigignenti-juglandacearum (Nair, Kostichka & Kuntz) Broders & Boland. This fungus causes branch and trunk cankers that can coalesce to girdle adult trees. Chapter 1 provides background information on butternut and butternut canker. We used next-generation sequencing to identify new nuclear DNA markers for butternut and Japanese walnut, a congener with which butternut readily hybridizes. We also examined the alignment of SSR repeat sequences in butternut and Japanese walnut with similar sequences from other angiosperms in public sequence databases. The methods used and results obtained in this process are detailed in Chapter 2. Chapter 3 summarizes an investigation of the environmental and genetic factors contributing to canker disease incidence, severity, and mortality in a large (n=113) population of butternut in southern Wisconsin and two other populations of butternut, one near the main study site in southern Wisconsin and another in the Great Smoky Mountains National Park. We present evidence for weak correlations of genetic similarity and phenotypic similarity for several disease traits, parentage analysis of regeneration in the smaller Wisconsin population, and evidence for significant microsite influences on butternut mortality over an 11-year period in the large Slocum's Woods butternut population.

Duchenne muscular dystrophies (DMD) and Limb-girdle muscular dystrophies (LGMD) are common inherited degenerative muscle diseases caused by mutations in genes coding for memberance associated proteins in muscle cells. DMD and LGMD often manifest themselves in young age and lead to severe morbidity and fatality, with no currently available effective treatment. In addition, the diseases are usually genotypically recessive, which makes them suitable for gene replacement therapy with vectors. Recombinant adeno-associated virus (rAAV) is one such vector based on defective human parvoviruses. rAAV system has attracted attention due to its non-pathogenicity, genomic integration, transduction of quiescent cells, and apparent lack of cellular immune reactions. In contrast to other viral vectors, rAAV is capable of efficiently bypassing the myofiber basal lamina and tranducing mature muscle cells. We have demonstrated that rAAV vectors harboring a foreign gene can achieve highly efficient and sustained gene transfer in mature muscle of immunocompetent animals for more than 1.5 years without detectable toxicity. Recently, significant improvement in vector production methodology has made it possible to generate high titer and high quality rAAV vectors completely free of helper adenovirus contamination. However, no experiments using rAAV vectors to restore the functional deficits in muscle tissue itself have been reported to date. In this proposal, we will use delta-SG as the target disease gene, the delta-SG deficient hamster as the LGDM animal model, and rAAV as the gene delivery vector to test our general hypothesis that safe, efficient and sustained functional rescue of muscle deficiency can be achieved by genetic complementation of inherited muscular dystrophies with rAAV vectors. Specifically, we would like to achieve the following aims: 1) To study gene transfer efficiency and functional rescue in the LGMD hamster model by local intramuscular delivery of delta-SG-rAAV vectors and examine their short term ability to correlate the genetic defect in both skeletal and cardiac muscle. 2) To evaluate the gene therapy efficacy after systemic delivery of rAAV vectors through intra-artery or intra-ventricle injection. 3) To investigate the molecular kinetics and fate of rAAV vectors, especially after systemic vector delivery. 4) To develop new generation, high titer, helper-virus-free rAAV producer cells, which not only harbor vector and packaging genes, but also contain the necessary helper genes from adenovirus in a highly regulated and inducible manner.

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