Computer implementation of the international standards for neurological classification of spinal cord injury for consistent and efficient derivation of its subscores including handling of data from not testable segments

The International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI), defined by the American Spinal Injury Association (ASIA), and particularly the ASIA Impairment Scale (AIS) are widely used for research and clinical purposes. Although detailed procedures for scaling, scoring, and classification have been defined, misclassifications remain a major problem, especially for cases with missing (i.e., not testable [NT]) data. This work aimed to implement computer-based classification algorithms that included rules for handling NT data. A consistent and structured algorithmic scoring, scaling, and classification scheme, and a computerized application have been developed by redefining logical/mathematical imprecisions. Existing scoring rules are extended for handling NT segments. Design criterion is a pure logical approach so that substitution of non-testability for all valid examination scores leads to concordant results. Nine percent of 5542 datasets from 1594 patients in the database of the European Multicenter Study of Human Spinal Cord Injury (EM-SCI) contained NT segments. After adjusting computational algorithms, the classification accuracy was equivalent between clinical experts and the computational approach and resulted in 84% valid AIS classifications within datasets containing NT. Additionally, the computational method is much more efficient, processing approximately 200,000 classifications/sec. Computational algorithms offer the ability to classify ISNCSCI subscores efficiently and without the risk of human-induced errors. This is of particular clinical relevance, since these scores are used for early predictions of neurological recovery and functional outcome for patients with spinal cord injuries

Homozygous expression of the myofibrillar myopathy-associated p.W2710X filamin C variant reveals major pathomechanisms of sarcomeric lesion formation

Filamin C (FLNc) is mainly expressed in striated muscle cells where it localizes to Z-discs, myotendinous junctions and intercalated discs. Recent studies have revealed numerous mutations in theFLNCgene causing familial and sporadic myopathies and cardiomyopathies with marked clinical variability. The most frequent myopathic mutation, p.W2710X, which is associated with myofibrillar myopathy, deletes the carboxy-terminal 16 amino acids from FLNc and abolishes the dimerization property of Ig-like domain 24. We previously characterized knock-in mice heterozygous for this mutation (p.W2711X), and have now investigated homozygous mice using protein and mRNA expression analyses, mass spectrometry, and extensive immunolocalization and ultrastructural studies. Although the latter mice display a relatively mild myopathy under normal conditions, our analyses identified major mechanisms causing the pathophysiology of this disease: in comparison to wildtype animals (i) the expression level of FLNc protein is drastically reduced; (ii) mutant FLNc is relocalized from Z-discs to particularly mechanically strained parts of muscle cells, i.e. myotendinous junctions and myofibrillar lesions; (iii) the number of lesions is greatly increased and these lesions lack Bcl2-associated athanogene 3 (BAG3) protein; (iv) the expression of heat shock protein beta-7 (HSPB7) is almost completely abolished. These findings indicate grave disturbances of BAG3-dependent and -independent autophagy pathways that are required for efficient lesion repair. In addition, our studies reveal general mechanisms of lesion formation and demonstrate that defective FLNc dimerization via its carboxy-terminal domain does not disturb assembly and basic function of myofibrils. An alternative, more amino-terminally located dimerization site might compensate for that loss. Since filamins function as stress sensors, our data further substantiate that FLNc is important for mechanosensing in the context of Z-disc stabilization and maintenance

Myofibrillar instability exacerbated by acute exercise in filaminopathy

Filamin C (FLNC) mutations in humans cause myofibrillar myopathy (MFM) and cardiomyopathy, characterized by protein aggregation and myofibrillar degeneration. We generated the first patient-mimicking knock-in mouse harbouring the most common disease-causing filamin C mutation (p.W2710X). These heterozygous mice developed muscle weakness and myofibrillar instability, with formation of filamin C- and Xin-positive lesions streaming between Z-discs. These lesions, which are distinct from the classical MFM protein aggregates by their morphology and filamentous appearance, were greatly increased in number upon acute physical exercise in the mice. This pathology suggests that mutant filamin influences the mechanical stability of myofibrillar Z-discs, explaining the muscle weakness in mice and humans. Re-evaluation of biopsies from MFM-filaminopathy patients with different FLNC mutations revealed a similar, previously unreported lesion pathology, in addition to the classical protein aggregates, and suggested that structures previously interpreted as aggregates may be in part sarcomeric lesions. We postulate that these lesions define preclinical disease stages, preceding the formation of protein aggregates

Size-related inbreeding preference and competitiveness in male Pelvicachromis taeniatus (Cichlidae)

Sexual selection is an important force in the evolution of body size. Both intersexual selection, that is, preference for large individuals, and intrasexual selection, that is, increased competitiveness of large individuals, are involved in this process. Furthermore, preferences based on body size of the choosing individual might also influence body size evolution. Here, we investigated male mate choice and competitiveness in relation to male body size in Pelvicachromis taeniatus, a size-dimorphic cichlid. In previous experiments, both sexes showed mating preferences for larger and genetically related individuals. First, we examined male inbreeding preferences based on olfactory cues. Males that highly varied in body size were given the choice between the odor of a familiar sister and the odor of an unfamiliar unrelated female that were presented in combination with a computer-animated image of a female P. taeniatus as a visual stimulus. Male preference for the odor of their sisters was correlated with male body size. Only larger males were choosy concerning related odors and preferred their sisters, whereas smaller males were unselective. Second, we showed that large males outcompete smaller males in contest over a breeding site. The extent of aggression was negatively correlated with the size difference between the 2 males. Variation in male choice may reflect an adaptive strategy: small, less competitive and less attractive males might avoid the risk of failing to mate at all by reducing choosiness. Consequently, only large competitive males should obtain the benefits of choice, which may further contribute to the selective advantages of large body size. Copyright 2011, Oxford University Press.

Overexpression of human BAG3P209L in mice causes restrictive cardiomyopathy

An amino acid exchange (P209L) in the HSPB8 binding site of the human co-chaperone BAG3 gives rise to severe childhood cardiomyopathy. To phenocopy the disease in mice and gain insight into its mechanisms, we generated humanized transgenic mouse models. Expression of human BAG3P209L-eGFP in mice caused Z-disc disintegration and formation of protein aggregates. This was accompanied by massive fibrosis resulting in early-onset restrictive cardiomyopathy with increased mortality as observed in patients. RNA-Seq and proteomics revealed changes in the protein quality control system and increased autophagy in hearts from hBAG3P209L-eGFP mice. The mutation renders hBAG3P209L less soluble in vivo and induces protein aggregation, but does not abrogate hBAG3 binding properties. In conclusion, we report a mouse model mimicking the human disease. Our data suggest that the disease mechanism is due to accumulation of hBAG3P209L and mouse Bag3, causing sequestering of components of the protein quality control system and autophagy machinery leading to sarcomere disruption

Identification of Xin-repeat proteins as novel ligands of the SH3 domains of nebulin and nebulette and analysis of their interaction during myofibril formation and remodeling

The Xin actin-binding repeat-containing proteins Xin and XIRP2 are exclusively expressed in striated muscle cells, where they are believed to play an important role in development. In adult muscle, both proteins are concentrated at attachment sites of myofibrils to the membrane. In contrast, during development they are localized to immature myofibrils together with their binding partner, filamin C, indicating an involvement of both proteins in myofibril assembly. We identify the SH3 domains of nebulin and nebulette as novel ligands of proline-rich regions of Xin and XIRP2. Precise binding motifs are mapped and shown to bind both SH3 domains with micromolar affinity. Cocrystallization of the nebulette SH3 domain with the interacting XIRP2 peptide PPPTLPKPKLPKH reveals selective interactions that conform to class II SH3 domain-binding peptides. Bimolecular fluorescence complementation experiments in cultured muscle cells indicate a temporally restricted interaction of Xin-repeat proteins with nebulin/nebulette during early stages of myofibril development that is lost upon further maturation. In mature myofibrils, this interaction is limited to longitudinally oriented structures associated with myofibril development and remodeling. These data provide new insights into the role of Xin actin-binding repeat-containing proteins (together with their interaction partners) in myofibril assembly and after muscle damage