![]() Recent studies have focused on the identification and characterization of pathogenic mutations in regulatory regions, including canonical RNA splice sites. Although more than 120 genes have been implicated in these diseases (, accessed April 2018) (Bonne, Rivier, & Hamroun, 2017), many affected patients remain undiagnosed despite extensive and costly genetic testing. This is especially true of inherited muscle diseases. The genetic etiologies of the remaining families often prove difficult to ascertain, as they may lie in unsequenced regions (for exomes and targeted sequence panels in particular) or may be cryptic mutations in regions that are already sequenced. Unfortunately, in many cohorts, NGS techniques identify pathogenic mutations in a minority of families (Lee et al., 2014 Yang et al., 2014) for neuromuscular disorders, in general, it has been found to be around 40% (Ankala et al., 2015), as well as for limb-girdle muscular dystrophy in particular (Ghaoui et al., 2015 Reddy et al., 2017, 2016). ![]() Next-generation sequencing (NGS) technologies are now routinely used to discover pathogenic mutations underlying rare genetic disorders. This study confirms the potential of novel in silico algorithms to detect cryptic mutations in existing NGS data expands the phenotypic spectrum of ATP2A1 mutations beyond classic Brody myopathy and suggests that genetic testing of ATP2A1 should be considered in patients with clinical myotonia. Ubiquitous knockdown of SERCA led to lethality in Drosophila, as did knockdown targeting differentiating or fusing myoblasts. Aberrant splicing of ATP2A1 was confirmed via qRT-PCR, and abnormal expression of the protein product sarcoplasmic/endoplasmic reticulum Ca ++ ATPase 1 (SERCA1) was demonstrated in quadriceps femoris tissue from the proband. The PANESS pipeline identified a homozygous ATP2A1 variant (NC_000016.9:g.28905928G>A NM_004320.4:c.1287G>A:p.(Glu429=)) that was predicted to cause the omission of exon 11. RNAi knockdown studies were performed in Drosophila to model the gene deficiency. Further analyses were performed via qRT-PCR, immunoblotting, and immunohistochemistry. We developed a bioinformatics pipeline that screens existing NGS data for potentially aberrant novel essential splice sites (PANESS) and performed a pilot study on a family with a myotonic disorder. Noncanonical splice sites are more difficult to ascertain. Standard variant analysis of next-generation sequence (NGS) data focuses on canonical splice sites. When transformations of marine organic matter are considered, differences in community composition and their different abilities to access organic matter should be taken into account.Pathogenic mutations causing aberrant splicing are often difficult to detect. The greater number of phylum- and subphylum-level lineages and operational taxonomic units in sediments than in seawater samples may reflect the necessity of a wider range of enzymatic capabilities and strategies to access organic matter that has already been degraded during passage through the water column. Thus, the broader enzymatic capabilities of the sedimentary microbial communities may result from the compositional differences between seawater and sedimentary microbial communities, rather than from gene expression differences among compositionally similar communities. To compare bacterial communities, 16S rRNA gene clone libraries were constructed from the same seawater and sediment samples they diverged strongly in composition. In seawater, in contrast, only 5 of the 7 polysaccharides and 2 of the 3 algal extracts were hydrolyzed, and hydrolysis rates in surface and deepwater were virtually identical. Sedimentary microbial communities hydrolyzed all of the fluorescently labeled polysaccharide and algal extracts, in most cases at higher rates in subsurface than surface sediments. Patterns of enzyme activities differed between seawater and sediments, not just quantitatively, in accordance with higher cell numbers in sediments, but also in their more diversified enzyme spectrum. communities to initiate organic matter degradation, we measured the extracellular enzymatic hydrolysis rates of 10 substrates (polysaccharides and algal extracts) in surface seawater and bottom water as well as in surface and anoxic sediments of an Arctic fjord. To compare the abilities of seawater and sedimentary microbial. Although carbon cycling and preservation depend critically on the capabilities of these microbial communities, their compositions and capabilities have seldom been examined simultaneously at the same site. Heterotrophic microbial communities in seawater and sediments metabolize much of the organic carbon produced in the ocean.
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