Investigations of peptides and proteases in green coffee beans

Cof. canephora var. robusta. The coffee types were not significantly different in peptide content, but were significantly different in peptide composition. Peptides with weakly acid pI values were mainly found in the Cof. robusta samples. In comparison, peptides from Cof. arabica samples ranged evenly from the weakly acidic up to the weakly basic pH range. The apparent molecular masses of the peptides in the two groups of samples lie between 4 kDa and 10 kDa. The cysteine content of the peptides was relatively high. Model roastings permitted the assumption that peptides make a contribution to coffee flavour. Peptides are already available in freshly harvested coffee. Model tests have suggested that processing after the coffee harvest has an influence on peptide composition but not on the amounts of peptide. All coffee samples contained some extractable proteases. The electrophoretically obtained enzyme patterns of Cof. arabica and Cof. robusta were different. Received: 6 September 1999     

Verformungsbezogene mittragende Breite niedriger Verbundträger

Effective width due to deflections of composite girders with shallow height. Due to the high slenderness of composite girders with shallow height their deflection behaviour is of great importance. In many cases the cross sectional dimensions are determined by the limitation of the deflections. In contrary to high composite beams the deflection behaviour of these girders is influenced especially by the bending state and cracking behaviour of the concrete slab. Among others the deflection or the stiffness of the composite girders depend on the effective width of the slab. This article reports on investigations of the deflection behaviour and the effective width of shallow composite girders taking into account the bending state and the cracking of the concrete slab. At first it is shown that for the calculation of stresses a different value of the effective width has to be taken into account than for the calculation of deflections. On basis of extensive experimental, analytical and numerical investigations the influences on the effective width are analysed. A wide‐ranging parametric study leads to the development of an approach of the effective width for the calculation of deflections and a proposal for a quasi‐elastic calculation of the deflections of shallow composite girders. At this it turns out that the effective width due to deflections is influenced above all by the bending state of the concrete slab, its cracking behaviour, its ratio width to length, the ratio of the individual stiffness components of the composite cross section and the load level. On basis of the proposed approach for the calculation of the effective width the deflections of shallow composite girders can be determined realistically. This leads to a more economic design of composite shallow girders than until now.     

Evaluating science arguments: Evidence, uncertainty, and argument strength.

Public debates about socioscientific issues are increasingly prevalent, but the public response to messages about, for example, climate change, does not always seem to match the seriousness of the problem identified by scientists. Is there anything unique about appeals based on scientific evidence—do people evaluate science and nonscience arguments differently? In an attempt to apply a systematic framework to people’s evaluation of science arguments, the authors draw on the Bayesian approach to informal argumentation. The Bayesian approach permits questions about how people evaluate science arguments to be posed and comparisons to be made between the evaluation of science and nonscience arguments. In an experiment involving three separate argument evaluation tasks, the authors investigated whether people’s evaluations of science and nonscience arguments differed in any meaningful way. Although some differences were observed in the relative strength of science and nonscience arguments, the evaluation of science arguments was determined by the same factors as nonscience arguments. Our results suggest that science communicators wishing to construct a successful appeal can make use of the Bayesian framework to distinguish strong and weak arguments. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Erhöhung der Ermüdungsfestigkeit von geschweißten höherfesten Baustählen durch Anwendung von Nachbehandlungsverfahren

Enhancement of the fatigue strength of welded high strength steels by application of post‐weld treatment methods. According to the present state of the art welded high strength steels have the same fatigue strength as welded standard steels. For an effective application of high strength steels in constructions subjected to fatigue are therefore additional efforts necessary in order to improve the fatigue strength, for example by the application of post‐weld treatment methods. However up to the present it is not possible to apply the positive effects of these methods in the fatigue design of steel structures. This paper shows first results of a research project, that examines the effectiveness of the post‐weld treatment method TIG‐dressing and the relatively new method “Ultrasonic Impact Treatment” (UIT) for the improvement of fatigue strength of welded high strength steels.     

A non-conserved sequence in the 5'region of the CYH2 intron from Saccharomyces cerevisiae controls splicing efficiency of the pre-mRNA

The CYH2 gene from Saccharomyces cerevisiae containing one 510 bp intron is spliced inefficiently. We have shown previously that a non-conserved sequence within the intron is responsible for this low splicing efficiency. Using synthetic oligonucleotides comprising the identified region we show in this report that a very short region contains the specificity to act negatively on the splicing efficiency of the CYH2 gene. Furthermore, this sequence influences the splicing efficiency only when it is placed close to the 5' splice site of the gene. Investigations with chimeric CYH2/beta-actin genes show that this sequence acts independent from its natural surroundings. We propose that this sequence might interact with splicing factor(s).     

Adaptation of Oxidative Phosphorylation Machinery Compensates for Hepatic Lipotoxicity in Early Stages of MAFLD

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron transport chain (ETC) in this pathologic condition is a consequence of shifted substrate availability. We addressed this question using a transgenic mouse model with increased hepatic insulin resistance and DNL due to constitutively active human SREBP-1c. The abundance of ETC complex subunits and components of key metabolic pathways are regulated in the liver of these animals. Further omics approaches combined with functional assays in isolated liver mitochondria and primary hepatocytes revealed that the SREBP-1c-forced fatty liver induced a substrate limitation for oxidative phosphorylation, inducing enhanced complex II activity. The observed increased expression of mitochondrial genes may have indicated a counteraction. In conclusion, a shift of available substrates directed toward activated DNL results in increased electron flows, mainly through complex II, to compensate for the increased energy demand of the cell. The reorganization of key compounds in energy metabolism observed in the SREBP-1c animal model might explain the initial increase in mitochondrial function observed in the early stages of human MAFLD.     

Structural Basis of Cyclic 1,3-Diene Forming Acyl-Coenzyme A Dehydrogenases

The biologically important, FAD-containing acyl-coenzyme A (CoA) dehydrogenases (ACAD) usually catalyze the anti-1,2-elimination of a proton and a hydride of aliphatic CoA thioesters. Here, we report on the structure and function of an ACAD from anaerobic bacteria catalyzing the unprecedented 1,4-elimination at C3 and C6 of cyclohex-1-ene-1-carboxyl-CoA (Ch1CoA) to cyclohex-1,5-diene-1-carboxyl-CoA (Ch1,5CoA) and at C3 and C4 of the latter to benzoyl-CoA. Based on high-resolution Ch1CoA dehydrogenase crystal structures, the unorthodox reactivity is explained by the presence of a catalytic aspartate base (D91) at C3, and by eliminating the catalytic glutamate base at C1. Moreover, C6 of Ch1CoA and C4 of Ch1,5CoA are positioned towards FAD-N5 to favor the biologically relevant C3,C6- over the C3,C4-dehydrogenation activity. The C1,C2-dehydrogenation activity was regained by structure-inspired amino acid exchanges. The results provide the structural rationale for the extended catalytic repertoire of ACADs and offer previously unknown biocatalytic options for the synthesis of cyclic 1,3-diene building blocks.     

A Multi-step Virtual Screening Protocol for the Identification of Novel Non-acidic Microsomal Prostaglandin E2 Synthase-1 (mPGES-1) Inhibitors

Microsomal prostaglandin E₂ synthase-1 (mPGES-1) is a potential therapeutic target for the treatment of inflammatory diseases and certain types of cancer. To identify novel scaffolds for mPGES-1 inhibition, we applied a virtual screening (VS) protocol that comprises molecular docking, fingerprints-based clustering with diversity-based selection, protein–ligand interactions fingerprints, and molecular dynamics (MD) simulations with molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) calculations. The hits identified were carefully analyzed to ensure the selection of novel scaffolds that establish stable interactions with key residues in the mPGES-1 binding pocket and inhibit the catalytic activity of the enzyme. As a result, we discovered two promising chemotypes, 4-(2-chlorophenyl)-N-[(2-{[(propan-2-yl)sulfamoyl]methyl}phenyl)methyl]piperazine-1-carboxamide ( 6 ) and N-(4-methoxy-3-{[4-(6-methyl-1,3-benzothiazol-2-yl)phenyl]sulfamoyl}phenyl)acetamide ( 8 ), as non-acidic mPGES-1 inhibitors with IC₅₀ values of 1.2 and 1.3 μm , respectively. Minimal structural optimization of 8 resulted in three more compounds with promising improvements in inhibitory activity (IC₅₀: 0.3–0.6 μm ). The unprecedented chemical structures of 6 and 8 , which are amenable to further derivatization, reveal a new and attractive approach for the development of mPGES-1 inhibitors with potential anti-inflammatory and anticancer properties.