CRISPR/Cas9-Based Mutagenesis of Histone H3.1 in Spinal Dynorphinergic Neurons Attenuates Thermal Sensitivity in Mice

Burn injury is a trauma resulting in tissue degradation and severe pain, which is processed first by neuronal circuits in the spinal dorsal horn. We have recently shown that in mice, excitatory dynorphinergic (Pdyn) neurons play a pivotal role in the response to burn-injury-associated tissue damage via histone H3.1 phosphorylation-dependent signaling. As Pdyn neurons were mostly associated with mechanical allodynia, their involvement in thermonociception had to be further elucidated. Using a custom-made AAV9_mutH3.1 virus combined with the CRISPR/cas9 system, here we provide evidence that blocking histone H3.1 phosphorylation at position serine 10 (S10) in spinal Pdyn neurons significantly increases the thermal nociceptive threshold in mice. In contrast, neither mechanosensation nor acute chemonociception was affected by the transgenic manipulation of histone H3.1. These results suggest that blocking rapid epigenetic tagging of S10H3 in spinal Pdyn neurons alters acute thermosensation and thus explains the involvement of Pdyn cells in the immediate response to burn-injury-associated tissue damage.     

Prenatal Exposure to Triclocarban Impairs ESR1 Signaling and Disrupts Epigenetic Status in Sex-Specific Ways as Well as Dysregulates the Expression of Neurogenesis- and Neurotransmitter-Related Genes in the Postnatal Mouse Brain

Triclocarban is a highly effective and broadly used antimicrobial agent. Humans are continually exposed to triclocarban, but the safety of prenatal exposure to triclocarban in the context of neurodevelopment remains unknown. In this study, we demonstrated for the first time that mice that had been prenatally exposed to environmentally relevant doses of triclocarban had impaired estrogen receptor 1 (ESR1) signaling in the brain. These mice displayed decreased mRNA and protein expression levels of ESR1 as well as hypermethylation of the Esr1 gene in the cerebral cortex. Prenatal exposure to triclocarban also diminished the mRNA expression of Esr2, Gper1, Ahr, Arnt, Cyp19a1, Cyp1a1, and Atg7, and the protein levels of CAR, ARNT, and MAP1LC3AB in female brains and decreased the protein levels of BCL2, ARNT, and MAP1LC3AB in male brains. In addition, exposure to triclocarban caused sex-specific alterations in the methylation levels of global DNA and estrogen receptor genes. Microarray and enrichment analyses showed that, in males, triclocarban dysregulated mainly neurogenesis-related genes, whereas, in females, the compound dysregulated mainly neurotransmitter-related genes. In conclusion, our data identified triclocarban as a neurodevelopmental risk factor that particularly targets ESR1, affects apoptosis and autophagy, and in sex-specific ways disrupts the epigenetic status of brain tissue and dysregulates the postnatal expression of neurogenesis- and neurotransmitter-related genes.     

Cyansäureester. 25 [1]. Umsetzung von Mercapto-N-heterocyclen mit Arylcyanaten

Cyanic Acid Esters. 25 The Reaction of Mercapto-N-heterocycles with Aryl Cyanates Unsaturated N-heterocycles containing a cyclic thio urea structure and aryl cyanates (molar ratio 1:2), react to form azolo(1,2-d) (1,2,4)thiadiazoles. This reaction has a wide scope concerning starting materials and gives good yields. Under the same conditions 2-thione-benzthiazolin is transformed to the corresponding disulfide. 4,5-Bis-mercaptomethylene-o-xylene and aryl cyanates give 2,3-dithia-6, 7-dimethyl-tetraline.     

Psyllium, not pectin or guar gum, alters lipoprotein and biliary bile acid composition and fecal sterol excretion in the hamster

Different soluble dietary fibers known to alter cholesterol metabolism were fed to golden Syrian hamsters, and their specific impact on lipoproteins, biliary bile acid profile, and fecal sterol excretion was evaluated. Semipurified diets containing 20% fat; 0.12% cholesterol; and 8% of psyllium (PSY); high (hePE) and low (lePE) esterified pectin; or high (hvGG) and low (lvGG) viscous guar gum were fed for 5 wk. Compared to control, PSY caused a significant reduction in plasma cholesterol (2.9±0.5 vs. 5.5±0.5 mmol/L), whereas hePE, lePE, hvGG, or lvGG had no apparent effect on plasma lipids. Hepatic total and esterified cholesterol were substantially decreased with PSY, pectin and guar gum, whereby PSY produced the most pronounced effect. Distinctive changes existed in the bile acid profile related to the different fibers. In contrast to pectin and guar gum, PSY caused a significant increase in the cholate:chenodeoxycholate and the glycine:taurine conjugation ratio. Pectin and guar gum did not alter daily fecal neutral sterol excretion while PSY caused a 90% increase due to a higher fecal output. Daily fecal bile acid excretion and total fecal bile acid concentration were significantly increased by PSY, whereas hePE, lePE, hvGG, and lvGG revealed no or only minor effects. Taken together, the disparate hypocholesterolemic effects of PSY, pectin, and guar gum on cholesterol and bile acid metabolism in the hamster are possibly related to different physicochemical properties, e.g., viscosity and susceptibility to fermentation, affecting the fiber-mediated action in the intestine.     

Customized Design of Scalable Microfluidic Droplet Generators Using Step‐Emulsification Methods

Customized monodisperse microparticles and microcapsules can be produced by combining microfluidic droplet generation with subsequent particle solidification. Established microfluidic devices for droplet formation like flow‐focusing structures or T‐junctions provide high reproducibility and controllability, but are often limited in terms of throughput or variability. A higher throughput by means of simple numbering‐up can be achieved by applying alternative droplet formation mechanisms like step emulsification. Using laser‐ablated glass reactors designed in‐house, comprehensive studies with varied step geometry and process parameters were performed in order to provide fundamental data for general calculation methods allowing the fast design of customized microfluidic droplet generators.     

Fused calcium zirconate for refractory applications

This article investigates dense CaZrO₃ produced by electric arc melting and compares it to solid state synthesized CaZrO₃. The fused CaZrO₃ had a considerable lower porosity than the solid state synthesized material, however due to evaporation of some CaO during the electric arc melting the stoichiometric ratio of CaO/ZrO₂ was slightly smaller than one, resulting in the formation of cubic ZrO₂ as a second phase. To determine the local phase distribution the materials were investigated with EBSD in addition to integral XRD. The vibration polishing of the ceramics resulted in excellent EBSD patterns. Die pressed cylinders of the fused material led to a porosity as low as 14.3%. The linear thermal expansion coefficient from 25 °C to 1000 °C was 10.45 × 10^?6 K^?1. The cold isostatic pressing of larger crucibles resulted in higher porosity of 15.9–16.5%. The EBSD analysis and the cold isostatic pressed crucibles can be applied in corrosion tests for titanium melts.     

The ɛ‐Amino Group of Protein Lysine Residues Is Highly Susceptible to Nonenzymatic Acylation by Several Physiological Acyl‐CoA Thioesters

Mitochondrial enzymes implicated in the pathophysiology of diabetes, cancer, and metabolic syndrome are highly regulated by acetylation. However, mitochondrial acetyltransferases have not been identified. Here, we show that acetylation and also other acylations are spontaneous processes that depend on pH value, acyl‐CoA concentration and the chemical nature of the acyl residue. In the case of a peptide derived from carbamoyl phosphate synthetase 1, the rates of succinylation and glutarylation were up to 150 times than for acetylation. These results were confirmed by using the protein substrate cyclophilin A (CypA). Deacylation experiments revealed that SIRT3 exhibits deacetylase activity but is not able to remove any of the succinyl groups from CypA, whereas SIRT5 is an effective protein desuccinylase. Thus, the acylation landscape on lysine residues might largely depend on the enzymatic activity of specific sirtuins, and the availability and reactivity of acyl‐CoA compounds.     

Impact of minimal heat-processing on pectin methylesterase and peroxidase activity in freshly squeezed <Emphasis Type="Italic">Citrus</Emphasis> juices

The impact of minimal heat-processing of juices on the activities of endogenous pectin methylesterase (PE) and peroxidase (POD) was compared between Citrus species. Mono-cultivar juices were produced from three orange (Citrus sinensis (L.) Osbeck cvs. ‘Navelina’, ‘Salustiana’, and ‘Navelate’), two lemon (Citrus limon (L.) Burm. f. cvs. ‘Verna’ and ‘Primofiori’), and two Clementine mandarin varieties (Citrus reticulata Blanco cvs. ‘Marisol’ and ‘Clemenules’). Two mandarin hybrids (cvs. ‘Ortanique’ and ‘Clemenvilla’) were likewise used. The freshly squeezed juices were subjected to continuous treatments at six different temperatures (42–92 °C) with subsequent re-cooling on the pilot plant scale. In fresh Citrus juice, POD activities notably varied between 0.2 and 7.5 nkat g⁻¹ of juice, whereas PE activities were more uniform (0.4–1.5 PE units g⁻¹). In all juices, except ‘Ortanique’ juice, heating ≥42 °C for 12 s reduced POD activity below 4.3% of the maximum activity in fresh Citrus juice. Thermal tolerance of the thermo-labile PE fraction was overall much higher, but varied among juices during heating at temperatures ≤62 °C. Overall thermal resistance of PE was though comparable, with deactivation exceeding 84%, mostly even 90%, after thermal treatments ≥72 °C. Unlike POD, total PE activity proved to be an indicator of freshness that is universally applicable to Citrus juices derived from orange, mandarin, and lemon or blends thereof. Freshly squeezed juices can analytically be distinguished from cold-stored, minimally processed products that display an almost completely inactivated thermo-labile PE fraction and thus extended shelf life.     

Textile-Based Sensors for Inspection of Composite Materials

The monitoring of mechanical deformation and damage of composite materials is normally performed by established analytical methods,such as strain gauges and opt...