Mice with endogenous TDP‐43 mutations exhibit gain of splicing function and characteristics of amyotrophic lateral sclerosis

TDP‐43 (encoded by the gene TARDBP) is an RNA binding protein central to the pathogenesis of amyotrophic lateral sclerosis (ALS). However, how TARDBP mutations trigger pathogenesis remains unknown. Here, we use novel mouse mutants carrying point mutations in endogenous Tardbp to dissect TDP‐43 function at physiological levels both in vitro and in vivo. Interestingly, we find that mutations within the C‐terminal domain of TDP‐43 lead to a gain of splicing function. Using two different strains, we are able to separate TDP‐43 loss‐ and gain‐of‐function effects. TDP‐43 gain‐of‐function effects in these mice reveal a novel category of splicing events controlled by TDP‐43, referred to as “skiptic” exons, in which skipping of constitutive exons causes changes in gene expression. In vivo, this gain‐of‐function mutation in endogenous Tardbp causes an adult‐onset neuromuscular phenotype accompanied by motor neuron loss and neurodegenerative changes. Furthermore, we have validated the splicing gain‐of‐function and skiptic exons in ALS patient‐derived cells. Our findings provide a novel pathogenic mechanism and highlight how TDP‐43 gain of function and loss of function affect RNA processing differently, suggesting they may act at different disease stages.     

The Ethics of Discharging Asylum Seekers to Harm: A Case From Australia

In February 2016 a twelve-month-old asylum seeker, who came to be know as Baby Asha, was transferred from Nauru and hospitalized in Brisbane. This case came to public attention after Doctors refused to discharge Asha as she would have been returned to detention on Nauru. What in other circumstances would have been considered routine clinical care, quickly turned into an act of civil disobedience. This paper will discuss the ethical aspects of this case, along with its implications for clinicians and the broader healthcare community.     

Efficacy of biopesticides on spotted wing drosophila,Drosophila suzukii Matsumura in fall red raspberries

Drosophila suzukii Matsumura is a significant pest of soft‐skinned fruit. Larvae of D. suzukii develop within the fruit making it unmarketable as fresh berries and increasing the risk of rejection by processors. We evaluated selected biopesticides for control of D. suzukii in fall red raspberries, Rubus idaeus L. The trial results highlight a small number of biopesticides with the potential to reduce infestation of Drosophila larvae in raspberries. In addition to the standard biopesticide spinosad, we found that sabadilla alkaloids and Chromobacterium subtsugae both reduced the number of Drosophila larvae in raspberry fruit. Treatments that included corn syrup as a feeding stimulant showed no significant difference in their infestation levels compared to treatments without the syrup. In the final week of the 5‐week trial, treatments with rotations of either spinosad/C. subtsugae or spinosad/sabadilla alkaloids had a 67% and 57% reduction in infestation when compared to untreated raspberries. Treatments of spinosad alone on a 7 day rotation and C. subtsugae alone on a 3–5 day rotation both had a 62% and 61% reduction in larval infestation when compared to untreated raspberries. Third instar larvae, the largest and most damaging, were significantly reduced in plots treated with spinosad only, a rotation of spinosad/sabadilla alkaloids and the rotation of spinosad/C. subtsugae with corn syrup added when compared to untreated plots. This suggests that either of these biopesticides could be used as effective rotation partners along with spinosad for control of D. suzukii. Our results highlight that biopesticides can provide significant reduction in this devastating pest when used alone or in combination, providing options to support resistance management.     

End-of-Life Infrastructure Economics for "Clean Vehicles" in the United States

Rising fuel prices and concern over emissions are prompting automakers and legislators to introduce and evaluate "clean vehicles" throughout the United States. Hybrid electric vehicles (HEVs) are now on the roads, electric vehicles (EVs) have been test marketed, and niche vehicles such as high-fuel-economy microcars are being considered for introduction. As these vehicles proliferate and mature, they will eventually reach their end of life (EOL). In the United States, an extensive recycling infrastructure exists for conventional, internal combustion engine (ICE) vehicles. Its primary constituents are the disassembler and the shredder. These industries, as well as battery recyclers, are expected to play integral roles in the EOL processing of clean vehicles.
A model of the automobile-recycling infrastructure and goal programming techniques are used to assess the materials streams and process profitabilities for several different clean vehicles. Two-seat EVs with lead-acid or NiMH batteries are compared with two- and four-seat HEVs and microcars. Changes to the nonferrous content in the vehicle bodies are explored and compared for the effect on processing profit-ability. Despite limitations associated with the linearity of goal programming techniques, application of this tool can still provide informative first-order results. Results indicate that although these clean vehicles may not garner the same profit levels as conventional ICE vehicles, they are profitable to process if there are markets for parts and if there are sufficient quantities of nonferrous materials.     

Brominated lipids identify lipid binding sites on the surface of the reaction center from Rhodobacter sphaeroides

This study describes the use of brominated phospholipids to distinguish between lipid and detergent binding sites on the surface of a typical alpha-helical membrane protein. Reaction centers isolated from Rhodobacter sphaeroides were cocrystallized with added brominated phospholipids. X-ray structural analysis of these crystals has revealed the presence of two lipid binding sites from the characteristic strong X-ray scattering from the bromine atoms. These results demonstrate the usefulness of this approach to mapping lipid binding sites at the surface of membrane proteins.     

Glyceraldehyde-3-phosphate dehydrogenase expression during apoptosis and proliferation of rat ventral prostate.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional enzyme known to play a critical role in neuronal apoptosis. We undertook the current studies to determine whether GAPDH also plays a role in prostate epithelial cell apoptosis in response to androgen deprivation. To do so, we analyzed GAPDH staining by immunohistochemistry during castration-induced involution and androgen-induced regeneration of rat ventral prostate. We found that GAPDH was undetectable in secretory epithelial cells at baseline and that staining did not increase in the epithelium during the period of peak apoptosis from 1 to 3 days after castration. However, GAPDH levels did increase within nuclei of some basal epithelial cells 5 days after castration and within the cytoplasm of all secretory epithelial cells 7 days after castration. GAPDH was also abundant within the cytoplasm of secretory epithelial cells during the period of maximal cell proliferation from 2 to 3 days after androgen replacement and was clearly apparent within nuclei of some epithelial cells 4 days after androgen replacement. Our studies suggest that GAPDH plays multiple roles during prostate epithelial cell apoptosis and proliferation.     

In Swedish families with hereditary prostate cancer, linkage to the HPC1 locus on chromosome 1q24-25 is restricted to families with early-onset prostate cancer.

Prostate cancer clusters in some families, and an estimated 5%-10% of all cases are estimated to result from inheritance of prostate cancer-susceptibility genes. We previously reported evidence of linkage to the 1q24-25 region (HPC1) in 91 North American and Swedish families each with multiple cases of prostate cancer (Smith et al. 1996). In the present report we analyze 40 (12 original and 28 newly identified) Swedish families with hereditary prostate cancer (HPC) that, on the basis of 40 markers spanning a 25-cM interval within 1q24-25, have evidence of linkage. In the complete set of families, a maximum two-point LOD score of 1.10 was observed at D1S413 (at a recombination fraction [theta] of.1), with a maximum NPL (nonparametric linkage) Z score of 1.64 at D1S202 (P=.05). The evidence of linkage to this region originated almost exclusively from the subset of 12 early-onset (age <65 years) families, which yielded a maximum LOD score of 2.38 at D1S413 (straight theta=0) and an NPL Z score of 1.95 at D1S422 (P=.03). Estimates from heterogeneity tests suggest that, within Sweden, as many as 50% of early-onset families had evidence of linkage to the HPC1 region. These results are consistent with the hypothesis of linkage to HPC1 in a subset of families with prostate cancer, particularly those with an early age at diagnosis.     

Structure-based design of novel groups for use in the P1 position of thrombin inhibitor scaffolds. Part 2: N-acetamidoimidazoles

Guided by X-ray crystallography of thrombin-inhibitor complexes and molecular modeling, alkylation of the N1 nitrogen of the imidazole P1 ligand of the pyridinoneacetamide thrombin inhibitor 1 with various acetamide moieties furnished inhibitors with significantly improved thrombin potency, trypsin selectivity, functional in vitro anticoagulant potency and in vivo antithrombotic efficacy. In the pyrazinoneacetamide series, oral bioavailability was also improved.