Identification of Six Novel SOD1 Gene Mutations in Familial Amyotrophic Lateral Sclerosis

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the premature death of motor neurons. In approximately 10% of the cases the disease is inherited as autosomal dominant trait (FALS). It has been found that mutations in the Cu/Zn superoxide dismutase gene (SODl) are responsible for approximately 15% of FALS kindreds. We screened affected individuals from 70 unrelated FALS kindreds and identified 10 mutations, 6 of which are novel. Surprisingly, we have found a mutation in exon 3, which includes most of the active site loop and Zn2+ binding sites, a region where no previous SOD1 mutations have been found. Our data increase the number of different SODl mutations causing FALS to 55, a significant fraction of the 154 amino acids of this relatively small protei

Variation in aggregation propensities among ALS-associated variants of SOD1: Correlation to human disease

To date, 146 different mutations in superoxide dismutase 1 (SOD1) have been identified in patients with familial amyotrophic lateral sclerosis (ALS). The mean age of disease onset in patients inheriting mutations in SOD1 is 45–47 years of age. However, although the length of disease duration is highly variable, there are examples of consistent disease durations associated with specific mutations (e. g. A4V, less than 2 years). In the present study, we have used a large set of data from SOD1-associated ALS pedigrees to identify correlations between disease features and biochemical/biophysical properties of more than 30 different variants of mutant SOD1. Using a reliable cell culture assay, we show that all ALS-associated mutations in SOD1 increase the inherent aggregation propensity of the protein. However, the relative propensity to do so varied considerably among mutants. We were not able to explain the variation in aggregation rates by differences in known protein properties such as enzyme activity, protein thermostability, mutation position or degree of change in protein charge. Similarly, we were not able to explain variability in the duration of disease in SOD1-associated ALS pedigrees by these properties. However, we find that the majority of pedigrees in which patients exhibit reproducibly short disease durations are associated with mutations that show a high inherent propensity to induce aggregation of SOD1

Induction of Cytoprotective Pathways Is Central to the Extension of Lifespan Conferred by Multiple Longevity Pathways

Many genetic and physiological treatments that extend lifespan also confer resistance to a variety of stressors, suggesting that cytoprotective mechanisms underpin the regulation of longevity. It has not been established, however, whether the induction of cytoprotective pathways is essential for lifespan extension or merely correlated. Using a panel of GFP-fused stress response genes, we identified the suites of cytoprotective pathways upregulated by 160 gene inactivations known to increase Caenorhabditis elegans longevity, including the mitochondrial UPR (hsp-6, hsp-60), the ER UPR (hsp-4), ROS response (sod-3, gst-4), and xenobiotic detoxification (gst-4). We then screened for other gene inactivations that disrupt the induction of these responses by xenobiotic or genetic triggers, identifying 29 gene inactivations required for cytoprotective gene expression. If cytoprotective responses contribute directly to lifespan extension, inactivation of these genes would be expected to compromise the extension of lifespan conferred by decreased insulin/IGF-1 signaling, caloric restriction, or the inhibition of mitochondrial function. We find that inactivation of 25 of 29 cytoprotection-regulatory genes shortens the extension of longevity normally induced by decreased insulin/IGF-1 signaling, disruption of mitochondrial function, or caloric restriction, without disrupting normal longevity nearly as dramatically. These data demonstrate that induction of cytoprotective pathways is central to longevity extension and identify a large set of new genetic components of the pathways that detect cellular damage and couple that detection to downstream cytoprotective effectors.National Institute on Aging (AG16636

PLP and MALDI-ToF determination of propagation rate coefficients of fast-polymerizing acrylates with heterocyclic side-chains: tetrahydrofurfuryl acrylate and (R)--acryloyloxy-β,β-dimethyl--butyrolactone

The PLP technique in combination with MALDI-ToF-MS was used to determine the Arrhenius plots for the propagation rate coefficients of THFA and ADBL, which polymerize much faster than alkyl acrylates. It is demonstrated that this is not due to higher propagation rate coefficients. It is shown that the temperature at which the PLP experiment brakes down increases in the order ADBL¿>¿THFA¿>¿alkyl acrylates, indicating a high extent of transfer to polymer for these monomers. Although kp decreases in the same order, the decrease of the corresponding overall polymerization rate is much larger. Therefore, the high polymerization rates of ADBL and THFA cannot be accounted for by fast propagation rates but more likely are linked to a low rate of terminatio

3D printing direct to industrial roll-to-roll casting for fast prototyping of scalable microfluidic systems.

Microfluidic technologies have enormous potential to offer breakthrough solutions across a wide range of applications. However, the rate of scale-up and commercialization of these technologies has lagged significantly behind promising breakthrough developments in the lab, due at least in part to the problems presented by transitioning from benchtop fabrication methods to mass-manufacturing. In this work, we develop and validate a method to create functional microfluidic prototype devices using 3D printed masters in an industrial-scale roll-to-roll continuous casting process. There were no significant difference in mixing performance between the roll-to-roll cast devices and the PDMS controls in fluidic mixing tests. Furthermore, the casting process provided information on the suitability of the prototype microfluidic patterns for scale-up. This work represents an important step in the realization of high-volume prototyping and manufacturing of microfluidic patterns for use across a broad range of applications

Ultraviolet–ultraviolet dual-cure process based on acrylate oxetane monomers

A dual-cure process consisting of two subsequent ultraviolet-initiated radical and cationic polymerizations was investigated. The process was studied with two acrylate oxetane monomers, one of them having a spacer between the two polymerizable moieties. It was shown by Fourier transform infrared (FTIR) that the first (radical) step was performed successfully for both systems. As for the second (cationic) step, only the monomer with the spacer was able to polymerize, allowing the crosslinking of the polyacrylic chains generated by the first step. The efficiency of the process was confirmed by differential scanning calorimetry because the glass-transition temperatures of the cured films were _16 and _ 34 °C after the first and second steps, respectively. The dual cure of this system was further analyzed by real-time FTIR, which showed that 86% of the acrylate and 80% of the oxetane moieties were converted after 20 and 50 s of light exposure, respectively

Dual-cure processes: towards deformable crosslinked coatings

Two dual-cure processes consisting of a UV-initiated radical polymerization followed by either a UV-induced cationic polymerisation, or a thermal addition reaction, were investigated. The feasibility of the processes was studied using an acrylate-oxetane monomer for the UV/UV combination, and an acrylated oligoester for the UV/Heat combination. It was shown by FTIR and Tg measurements, that both steps of each process could be performed efficiently and separately. This allowed the production of a deformable partially cured coating, whose cure can then be completed, leading to the required final properties. Furthermore, it was demonstrated that the increase of the functionality of the reactive diluent led to a decrease of the thermal crosslinking extent. This is probably due to the reduced mobility of the reactive species that is caused by an enhanced UV crosslinking taking place during the first step