Alpha Synuclein only Forms Fibrils In Vitro when Larger than its Critical Size of 70 Monomers

The aggregation of α-synuclein into small soluble aggregates and then fibrils is important in the development and spreading of aggregates through the brain in Parkinson's disease. Fibrillar aggregates can grow by monomer addition and then break into fragments that could spread into neighboring cells. The rate constants for fibril elongation and fragmentation have been measured but it is not known how large an aggregate needs to be before fibril formation is thermodynamically favorable. This critical size is an important parameter controlling at what stage in an aggregation reaction fibrils can form and replicate. We determined this value to be approximately 70 monomers using super-resolution and atomic force microscopy imaging of individual α-synuclein aggregates formed in solution over long time periods. This represents the minimum size for a stable α-synuclein fibril and we hypothesis the formation of aggregates of this size in a cell represents a tipping point at which rapid replication occurs.     

Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites

Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L⁻¹). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.     

Extracting safe thread schedules from incomplete model checking results

Model checkers frequently fail to completely verify a concurrent program, even if partial-order reduction is applied. The verification engineer is left in doubt whether the program is safe and the effort toward verifying the program is wasted. We present a technique that uses the results of such incomplete verification attempts to construct a (fair) scheduler that allows the safe execution of the partially verified concurrent program. This scheduler restricts the execution to schedules that have been proven safe (and prevents executions that were found to be erroneous). We evaluate the performance of our technique and show how it can be improved using partial-order reduction. While constraining the scheduler results in a considerable performance penalty in general, we show that in some cases our approach—somewhat surprisingly—even leads to faster executions.

     

Mammalian-Like Inflammatory and Pro-Resolving Oxylipins in Marine Algae

Oxylipins constitute a family of oxidized fatty acids, that are well known as tissue hormones in mammals. They contribute to inflammation and its resolution. The major classes of these lipid mediators are inflammatory prostaglandins (PGs) and leukotrienes (LTs) as well as pro-resolving resolvins (Rvs). Understanding their biosynthetic pathways and modes of action is important for anti-inflammatory interventions. Besides mammals, marine algae also biosynthesize mammalian-like oxylipins and thus offer new opportunities for oxylipin research. They provide prolific sources for these compounds and offer unique opportunities to study alternative biosynthetic pathways to the well-known lipid mediators. Herein, we discuss recent findings on the biosynthesis of oxylipins in mammals and algae including an alternative pathway to prostaglandin E₂, a novel pathway to a precursor of leukotriene B₄, and the production of resolvins in algae. We evaluate the pharmacological potential of the algal metabolites with implications in health and disease.     

MidDRIFTS-PLSR-based quantification of physico-chemical soil properties across two agroecological zones in Southwest Germany: generic independent validation surpasses region specific cross-validation

Development of spectroscopic prediction models via partial least squares regression (PLSR) suggests that model performance is highly affected by means of calibration and nature of the dataset. This study compares the predictive performance of PLSR models obtained by cross-validation and independent validation to quantify physico-chemical soil properties from their mid-infrared diffuse reflectance Fourier transform spectra (midDRIFTS) across two contrasting regions, Kraichgau (K) and Swabian Alb (SA), in Southwest Germany. We evaluated the capability of midDRIFTS-PLSR models for predicting total carbon (TC), organic carbon (TOC), inorganic carbon (TIC), nitrogen (TN), mineral N (N_min), C:N ratio, hot water extractable C and N (C_HWE, N_HWE), microbial biomass C and N (C_mic, N_mic), pH, bulk density, and clay, silt and sand contents of 126 soil samples. Based on calibrated models, most soil properties were predicted successfully using either calibration approach with residual prediction deviations ≥3 and R² > 0.9, except for N_min, C/N ratio, pH, bulk density and sand. However, predictive performance of generic independent validation derived models (GIC) of test set was considerably higher than generic cross-validation models. Validation using GIC models gave relatively the same predictive performance with those obtained in calibration except for N_min. Validation of region specific cross-validated models, however, resulted in successful predictions only for TC, TIC, TOC and TN in SA and TC and TIC and TOC in K. Our results show the superiority of independent validation over both generic and region specific cross-validation as a robust tool for predicting soil properties without further laboratory measurements.     

Stratification Dynamics and Geothermal Potential of a Deep Shaft in the Flooded Wolf Mine,Siegerland/Germany

Mine Water and the Environment - Mine water hydraulics and geothermal potential of a deep shaft of the flooded Wolf mine in the Siegerland ore district of the Rhenish Massif in Germany were...     

N-Substituted Nipecotic Acids as (S)-SNAP-5114 Analogues with Modified Lipophilic Domains

Potential mGAT4 inhibitors derived from the lead substance (S)-SNAP-5114 have been synthesized and characterized for their inhibitory potency. Variations from the parent compound included the substitution of one of its aromatic 4-methoxy and 4-methoxyphenyl groups, respectively, with a more polar moiety, including a carboxylic acid, alcohol, nitrile, carboxamide, sulfonamide, aldehyde or ketone function, or amino acid partial structures. Furthermore, it was investigated how the substitution of more than one of the aromatic 4-methoxy groups affects the potency and selectivity of the resulting compounds. Among the synthesized test substances (S)-1-{2-[(4-formylphenyl)bis(4-methoxyphenyl)-methoxy]ethyl}piperidine-3-carboxylic acid, that features a carbaldehyde function in place of one of the aromatic 4-methoxy moieties of (S)-SNAP-5114, was found to have a pIC₅₀ value of 5.89±0.07, hence constituting a slightly more potent mGAT4 inhibitor than the parent substance while showing comparable subtype selectivity.