Production and characterization of murine models of classic and intermediate maple syrup urine disease

BACKGROUND: Maple Syrup Urine Disease (MSUD) is an inborn error of metabolism caused by a deficiency of branched-chain keto acid dehydrogenase. MSUD has several clinical phenotypes depending on the degree of enzyme deficiency. Current treatments are not satisfactory and require new approaches to combat this disease. A major hurdle in developing new treatments has been the lack of a suitable animal model. METHODS: To create a murine model of classic MSUD, we used gene targeting and embryonic stem cell technologies to create a mouse line that lacked a functional E2 subunit gene of branched-chain keto acid dehydrogenase. To create a murine model of intermediate MSUD, we used transgenic technology to express a human E2 cDNA on the knockout background. Mice of both models were characterized at the molecular, biochemical, and whole animal levels. RESULTS: By disrupting the E2 subunit gene of branched-chain keto acid dehydrogenase, we created a gene knockout mouse model of classic MSUD. The homozygous knockout mice lacked branched-chain keto acid dehydrogenase activity, E2 immunoreactivity, and had a 3-fold increase in circulating branched-chain amino acids. These metabolic derangements resulted in neonatal lethality. Transgenic expression of a human E2 cDNA in the liver of the E2 knockout animals produced a model of intermediate MSUD. Branched-chain keto acid dehydrogenase activity was 5–6% of normal and was sufficient to allow survival, but was insufficient to normalize circulating branched-chain amino acids levels, which were intermediate between wildtype and the classic MSUD mouse model. CONCLUSION: These mice represent important animal models that closely approximate the phenotype of humans with the classic and intermediate forms of MSUD. These animals provide useful models to further characterize the pathogenesis of MSUD, as well as models to test novel therapeutic strategies, such as gene and cellular therapies, to treat this devastating metabolic disease

The etiology of schizophrenia and the origin of language: overview of a theory.

Schizophrenia is present in all human populations with approximately the same incidence. Why does such illness persist given that it is associated with a reproductive disadvantage? What is the balancing advantage? A possible explanation is linked to human language. According to this hypothesis schizophrenia occurs as a manifestation of genetic diversity associated with language--the function by which Homo sapiens has separated from other primate species. Language originated by a genetic mutation that allowed the cerebral hemispheres to develop with a degree of specialization (or lateralization) reflected in cerebral asymmetries. Individuals with schizophrenia show lesser structural and functional brain asymmetries than the population as a whole, and this finding can be interpreted as a delay, or failure in, establishing hemispheric dominance for language. We review recent evidence supporting this theory

The etiology of schizophrenia and the origin of language: overview of a theory.

Schizophrenia is present in all human populations with approximately the same incidence. Why does such illness persist given that it is associated with a reproductive disadvantage? What is the balancing advantage? A possible explanation is linked to human language. According to this hypothesis schizophrenia occurs as a manifestation of genetic diversity associated with language--the function by which Homo sapiens has separated from other primate species. Language originated by a genetic mutation that allowed the cerebral hemispheres to develop with a degree of specialization (or lateralization) reflected in cerebral asymmetries. Individuals with schizophrenia show lesser structural and functional brain asymmetries than the population as a whole, and this finding can be interpreted as a delay, or failure in, establishing hemispheric dominance for language. We review recent evidence supporting this theory

When epitaxy meets plasma: a path to ordered nanosheets arrays

The possibility of a controlled assembly of 2-dimensional (2D) nanosheets (NSs) into ordered arrays or even more sophisticated structures offers tremendous opportunities in the context of fabrication of a variety of NSs based devices. Reports of such ordered NSs are rare and all conventional “top-down” methods typically led to coarse structures exhibiting only limited surface quality. In this work, we demonstrate a path to directly synthesis ordered NSs arrays in a plasma activated chemical vapor deposition technique utilizing planar defects formed during hetero-epitaxial growth of crystals featuring a close-packed lattice. As an example, the synthesis of 3C-SiC NSs arrays with well-defined orientation on (001) and (111) Si substrates is shown. A detailed analysis identifies planar defects and the plasma environment as key factors determining the resulting 2D NSs arrays. Consequently, a “planar defects induced selective growth” effect is proposed to elucidate the corresponding growth mechanism

Expanding the druggable space of the LSD1/CoREST epigenetic target: New potential binding regions for drug-like molecules, peptides, protein partners, and chromatin

Lysine specific demethylase-1 (LSD1/KDM1A) in complex with its corepressor protein CoREST is a promising target for epigenetic drugs. No therapeutic that targets LSD1/CoREST, however, has been reported to date. Recently, extended molecular dynamics (MD) simulations indicated that LSD1/CoREST nanoscale clamp dynamics is regulated by substrate binding and highlighted key hinge points of this large-scale motion as well as the relevance of local residue dynamics. Prompted by the urgent need for new molecular probes and inhibitors to understand LSD1/CoREST interactions with small-molecules, peptides, protein partners, and chromatin, we undertake here a configurational ensemble approach to expand LSD1/CoREST druggability. The independent algorithms FTMap and SiteMap and our newly developed Druggable Site Visualizer (DSV) software tool were used to predict and inspect favorable binding sites. We find that the hinge points revealed by MD simulations at the SANT2/Tower interface, at the SWIRM/AOD interface, and at the AOD/Tower interface are new targets for the discovery of molecular probes to block association of LSD1/CoREST with chromatin or protein partners. A fourth region was also predicted from simulated configurational ensembles and was experimentally validated to have strong binding propensity. The observation that this prediction would be prevented when using only the X-ray structures available (including the X-ray structure bound to the same peptide) underscores the relevance of protein dynamics in protein interactions. A fifth region was highlighted corresponding to a small pocket on the AOD domain. This study sets the basis for future virtual screening campaigns targeting the five novel regions reported herein and for the design of LSD1/CoREST mutants to probe LSD1/CoREST binding with chromatin and various protein partners