Novel Furin Inhibitors with Potent Anti‐infectious Activity

New peptidomimetic furin inhibitors with unnatural amino acid residues in the P3 position were synthesized. The most potent compound 4‐guanidinomethyl‐phenylacteyl‐Arg‐Tle‐Arg‐4‐amidinobenzylamide (MI‐1148) inhibits furin with a K_i value of 5.5 pM . The derivatives also strongly inhibit PC1/3, whereas PC2 is less affected. Selected inhibitors were tested in cell culture for antibacterial and antiviral activity against infectious agents known to be dependent on furin activity. A significant protective effect against anthrax and diphtheria toxin was observed in the presence of the furin inhibitors. Furthermore, the spread of the highly pathogenic H5N1 and H7N1 avian influenza viruses and propagation of canine distemper virus was strongly inhibited. Inhibitor MI‐1148 was crystallized in complex with human furin. Its N‐terminal guanidinomethyl group in the para position of the P5 phenyl ring occupies the same position as that found previously for a structurally related inhibitor containing this substitution in the meta position, thereby maintaining all of the important P5 interactions. Our results confirm that the inhibition of furin is a promising strategy for a short‐term treatment of acute infectious diseases.     

Systematische Untersuchung der Azikularferritbildung in Stählen

The formation of acicular ferrite is mainly influenced by the steel composition, cooling rate, austenite grain size, and inclusion landscape. However, their precise effects have not been clarified in literature yet. Within the FWF project TRP 266-N19, a systematic methodology to study the influencing factors of the acicular ferrite formation as well as their interactions on laboratory scale was developed. The present work exemplarily demonstrates the application of this methodology for studying the effect of cooling rate and boron on the formation of acicular ferrite in a low-carbon HSLA steel.     

Fibroblast Growth Factor 21 Response in a Preclinical Alcohol Model of Acute-on-Chronic Liver Injury

Background and Aims: Fibroblast growth factor (FGF) 21 has recently been shown to play a potential role in bile acid metabolism. We aimed to investigate the FGF21 response in an ethanol-induced acute-on-chronic liver injury (ACLI) model in Abcb4^−/− mice with deficiency of the hepatobiliary phospholipid transporter. Methods: Total RNA was extracted from wild-type (WT, C57BL/6J) and Abcb4⁻/⁻ (KO) mice, which were either fed a control diet (WT-Cont and KO-Cont groups; n = 28/group) or ethanol diet, followed by an acute ethanol binge (WT-EtOH and KO-EtOH groups; n = 28/group). A total of 58 human subjects were recruited into the study, including patients with alcohol-associated liver disease (AALD; n = 31) and healthy controls (n = 27). The hepatic and ileal expressions of genes involved in bile acid metabolism, plasma FGF levels, and bile acid and its precursors 7α- and 27-hydroxycholesterol (7α- and 27-OHC) concentrations were determined. Primary mouse hepatocytes were isolated for cell culture experiments. Results: Alcohol feeding significantly induced plasma FGF21 and decreased hepatic Cyp7a1 levels. Hepatic expression levels of Fibroblast growth factor receptor 1 (Fgfr1), Fgfr4, Farnesoid X-activated receptor (Fxr), and Small heterodimer partner (Shp) and plasma FGF15/FGF19 levels did not differ with alcohol challenge. Exogenous FGF21 treatment suppressed Cyp7a1 in a dose-dependent manner in vitro. AALD patients showed markedly higher FGF21 and lower 7α-OHC plasma levels while FGF19 did not differ. Conclusions: The simultaneous upregulation of FGF21 and downregulation of Cyp7a1 expressions upon chronic plus binge alcohol feeding together with the invariant plasma FGF15 and hepatic Shp and Fxr levels suggest the presence of a direct regulatory mechanism of FGF21 on bile acid homeostasis through inhibition of CYP7A1 by an FGF15-independent pathway in this ACLI model. Lay Summary: Alcohol challenge results in the upregulation of FGF21 and repression of Cyp7a1 expressions while circulating FGF15 and hepatic Shp and Fxr levels remain constant both in healthy and pre-injured livers, suggesting the presence of an alternative FGF15-independent regulatory mechanism of FGF21 on bile acid homeostasis through the inhibition of Cyp7a1.     

Nitritation performance in membrane-aerated biofilm reactors differs from conventional biofilm systems

Nitrogen removal via nitrite has gained increasing attention in recent years due to its potential cost savings. Membrane-aerated biofilm reactors (MABRs) are one potential technology suitable to achieve nitritation. In this study we compared lab scale MABRs with conventional biofilm reactors to evaluate the influence of environmental conditions and operational parameters on nitritation performance. The oxygen mass transfer rate is postulated as a crucial parameter to control nitritation in the MABR: Clean water measurements showed significant underestimation of the total oxygen mass transfer, however, accurate determination of the oxygen mass transfer coefficient (k_m) of the system could be achieved by adjusting the liquid-phase mass transfer resistance in the constructed model. Batch experiments at different initial ammonium concentrations revealed that the conventional biofilm geometry was superior for nitritation compared to MABRs. These differences were reflected well in estimates of the oxygen affinity constants of the key microbial players, AOB and NOB (K_O,AOB < K_O,NOB (in both systems) and K_O,NOB values smaller in the MABR vs. the conventional biofilm system). It also appeared that – in addition to oxygen limitation – the absolute and relative substrate concentrations in the biofilm (esp. of oxygen) are very important for successful nitritation. Initial biomass composition, furthermore, impacted reactor performance in the MABR systems indicating the need for appropriate inoculum choice.     

Probing enzyme promiscuity of SGNH hydrolases

Several hydrolases of the SGNH superfamily, including the lipase SrLip from Streptomyces rimosus (Q93MW7), the acyl-CoA thioesterase I TesA from Pseudomonas aeruginosa (Q9HZY8) and the two lipolytic enzymes EstA (from P. aeruginosa, O33407) and EstP (from Pseudomonas putida, Q88QS0), were examined for promiscuity. These enzymes were tested against four chemically different classes of a total of 34 substrates known to be hydrolysed by esterases, thioesterases, lipases, phospholipases, Tweenases and proteases. Furthermore, they were also analysed with respect to their amino acid sequences and structural homology, and their phylogenetic relationship was determined. The Pseudomonas esterases EstA and EstP each have an N-terminal domain with catalytic activity together with a C-terminal autotransporter domain, and so the hybrid enzymes EstA(N)-EstP(C) and EstP(N)-EstA(C) were constructed by swapping the corresponding N- and C-terminal domains, and their hydrolytic activities were compared. Interestingly, substrate specificity and kinetic measurements indicated a significant influence of the autotransporter domains on the catalytic activities of these enzymes in solution. TesA, EstA and EstP were shown to function as esterases with different affinities and catalytic efficacies towards p-nitrophenyl butyrate. Of all the enzymes tested, only SrLip revealed lipase, phospholipase, esterase, thioesterase and Tweenase activities.     

Development of compatible wet-clean stripper for integration of CoWP metal cap in Cu/low-k interconnects

Implementation of CoWP metal caps into Cu/low-k integration schemes requires a wet stripper that not only gives efficient cleaning but also has good compatibility to CoWP and low-k dielectrics. This paper describes a novel non-fluoride CoWP compatible stripper, developed based on a systematic study of the effect of stripper components, i.e. solvent, corrosion inhibitor, and stripper pH. Electrochemical methods were used to characterize galvanic corrosion of the CoWP/Cu couple and to estimate CoWP etch rate. Our studies showed that a traditional fluoride stripper caused severe damage to CoWP capping layer. The new stripper achieved a good balance between cleaning efficiency and compatibility to CoWP and low-k dielectrics, and demonstrated significant advantages in electrical properties over the traditional fluoride stripper.     

Mercury contamination in the vicinity of a derelict chlor-alkali plant Part II: contamination of the aquatic and terrestrial food chain and potential risks to the local population

This study investigated the environmental impact and level of risk associated with mercury (Hg) contamination near a derelict chlor-alkali plant in Pavlodar, Northern Kazakhstan. Several species of fish were sampled from the highly polluted Lake Balkyldak and the nearby river Irtysh, to assess the extent of Hg bioaccumulation in the aquatic food chain and potential human health risks. A small number of bovine tissue samples, water samples, soil and plant samples from a nearby village were also investigated in order to make a preliminary assessment of potential impacts on the terrestrial food chain. Mercury levels in fish caught from Lake Balkyldak ranged from 0.16 to 2.2 mg kg(-1) and the majority of fish exceeded current human health criteria for Hg. Interspecies comparisons indicated that Hg is accumulated in the order dace>carp>tench. Site-specific bioaccumulation factors (BAF) were calculated for THg, and were estimated for MeHg. Fish from the river Irtysh and floodplain oxbow lakes contained between 0.075 and 0.159 mg kg(-1) of Hg and can be regarded as uncontaminated. Soils were found to be impacted by past atmospheric emissions of Hg. Cattle grazing in the surroundings of the factory are exposed to Hg from contaminated soils, plants and surface water, but the consumption of contaminated fish from the lake appears to be the main route of exposure for humans.     

Mercury contamination in the vicinity of a derelict chlor-alkali plant. Part I: sediment and water contamination of Lake Balkyldak and the River Irtysh

A mercury-cell chlor-alkali plant operated in Pavlodar, Northern Kazakhstan, for 18 years and caused widespread contamination of the surrounding environment. Untreated wastewater from the plant was discharged to Lake Balkyldak, a shallow impounded lake without an outlet. The nearby River Irtysh was also suspected to be impacted by mercury (Hg) via the transport of contaminated groundwater. We took sediment and water samples from both aquatic systems, and also sampled soils along the shoreline of the lake and in the Irtysh flood plain. Sediments from Lake Balkyldak were found to be very heavily contaminated, with Hg concentrations in the surface layer reaching up to approximately 1500 mg kg(-1) near the wastewater outfall pipe. The contaminated lake sediments are prone to wind-driven resuspension and are acting as a strong source of Hg to the water column. Unfiltered lake water samples taken in shallow areas within 10-15 m from the shoreline contained from 0.11 microg Hg L(-1) in the less contaminated northern part of the lake to 1.39 microg L(-1) near the pollutant outfall in the south (up to 7.3 microg L(-1) on windy days). Sediments from the River Irtysh were only slightly impacted, with maximum Hg concentrations of 0.046 mg kg(-1) in the old river channel and 0.36 mg kg(-1) in floodplain oxbow lakes. In water samples from the River Irtysh, Hg was generally not detected, although trace concentrations (3 to 9 ng L(-1)) were found in some samples taken from oxbow lakes. We conclude that the river is not significantly impacted by Hg, but the highly contaminated Lake Balkyldak poses a threat and is in need of remediation. Potential remediation options for the lake are reviewed and are discussed in the context of experiences made at other Hg-contaminated sites.     

Prof. Dr.-Ing. Dr. h. c. Franz Kollmann Zum 65. Geburtstag

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