Influence of gender on chronic ethanol-induced alterations in GABAA receptors in rats

Ethanol dependence, arising from chronic ethanol exposure, is associated with neuroadaptations of GABAA receptors, evidenced by alterations in various behaviors, receptor responsiveness and subunit gene expression. The present studies explored the effects of ethanol dependence in female rats for comparison with previous studies in our laboratory using male rats. We found that ethanol dependence resulted in differential effects on GABAA receptor gene expression in female rat cerebral cortex compared to ethanol dependent male rats. Notably, chronic ethanol consumption did not change GABAA receptor alpha 1 subunit peptide levels in ethanol dependent female rat cortex, in contrast to previously observed decreases in alpha 1 subunit expression in ethanol dependent male rat cortex. The effects of ethanol dependence on additional GABAA receptor subunit peptide levels (alpha 4, beta 2/3 and gamma 2) were similar, but not identical, between female and male rat cortex. When directly compared within the experiment, male and female rats had similar baseline bicuculline seizure thresholds and displayed a similar increase in seizure susceptibility during ethanol withdrawal. Ethanol withdrawn female rats were cross tolerant to the anticonvulsant effects of diazepam, similar to the findings in ethanol withdrawn male rats. Ethanol withdrawn female rats showed a dose-dependent enhancement of the anticonvulsant effect of the neuroactive steroid, THDOC (3 alpha,21-dihydroxy-5 alpha-pregnan-20-one) compared to control animals. This finding is similar to previous observations of increased sensitivity to the anticonvulsant effect of 3 alpha,5 alpha-THP (3 alpha-hydroxy-5 alpha-pregnan-20-one) in ethanol withdrawn male and female rats. In addition, low dose administration of THDOC elevated seizure thresholds in ethanol withdrawn female but not male rats, suggesting that ethanol withdrawn female rats were more responsive to the anticonvulsant effects of this neurosteroid than were ethanol withdrawn male rats. These findings show that gender impacts on adaptations in GABAA receptors elicited by ethanol dependence. However, the physiological outcomes of the differential alterations are not clear. Taken together, these studies suggest that additional mechanisms, beyond effects on GABAA receptor gene expression are involved in the mediation of ethanol dependence and withdrawal.     

Co-oxidation of carotene and crocin by soyabean lipoxygenase isoenzymes

Summary Three lipoxygenases that occur in soya beans were separated chromatographically. L-1 (optimum pH = 9.0), L-2 (pH 6.5), L-3 (pH 6.5). The velocities with which these enzymes co-oxidise-carotene or Crocin in the presence of linoleic acid or linoleyl sulphate weremeasured. The carotenoid turnover was related to each lipoxygenase activity.-carotene/linoleic acid = 55% (L-2), 43% (L-3), 6% (L-1), Crocin/linoleic acid = 17,8% (L-2), 14,3% (L-3), 3.3% (L-1), crocin/linoleyl sulphate = 24% (L-3), 4,2% (L-1).The relationship between the reaction rate of the Crocin bleaching and the concentrations of the enzyme, Crocin and linoleic acid was determined. To explain the differences between the pH-6.5 (L-2, L-3) and the alkaline (L-1) lipoxygenases it is supposed that L-2 and L-3 form specially active radicals that are able to co-oxidise polyenes. Both enzymes possess a hydrophobic bonding position, in the neighbourhood of the active site, for-carotene.

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Co-oxidation von Carotin und Crocin durch Lipoxygenase-Isoenzyme aus der Sojabohne

Zusammenfassung Drei in Sojabohnen vorkommende Lipoxygenasen wurden chromatographisch getrennt: L-1 (pH-Optimum 9,0), L-2 (pH 6,5), L-3 (pH 6,5). Gemessen wurden die Geschwindigkeiten mit denen diese Enzyme-Carotin oder Crocin in Gegenwart von Linolsäure oder Linoleylsulfat co-oxydieren. Die Carotinoid-Umsätze wurden auf die jeweilige Lipoxygenase-Aktivität bezogen.-Carotin/Linolsäure: 55% (L-2), 43% (L-3), 6% (L-1), Crocin/Linolsäure: 17,8% (L-2),14,3% (L-3),3,3% (L-1), Crocin/Linoleylsulfat: 24% (L-3), 4,2% (L-1).Für die Crocin-Bleichung wurde die Abhängigkeit der Reaktionsgeschwindigkeit von der Enzym-, Crocin- und Linolsäure-Konzentration bestimmt.Zur Erklärung der Unterschiede zwischen den pH 6,5- (L-2, L-3) und der alkalischen Lipoxygenase (L-1) wird angenommen: L-2 und L-3 bilden besonders aktiv Radikale, welche die Polyene co-oxydieren können. Beide Enzyme besitzen in der Nähe des aktiven Zentrums eine hydrophobe Bindungsstelle für das-Carotin.

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We are grateful to the Deutsche Forschungsgemeinschaft Bonn-Bad Godesberg for supporting this work.     

Furan fatty acid concentrations in tea infusions prepared from green,black, and herbal teas

Furan fatty acids (FuFAs) are valuable antioxidants with highly effective radical scavenging properties which are widely distributed at low levels in food. Previous research indicated that tea is a valuable source of FuFAs. However, tea is only consumed in form of infusions. To fill this gap, we prepared infusions from different herbal, green, and black teas. Initial measurements with GC-MSMS of tea verified previous findings that 11-(3,4-dimethyl-5-penylfuran-2-yl)-undecanoic acid (11D5) was the prevalent FuFA in tea matrix. Therefore, 11D5 was quantified in tea infusions by means of UHPLC-MSMS equipment after mild alkaline hydrolysis. While herbal tea infusions were low or free of FuFAs, 11D5 was detectable in all samples of green and black tea infusions. Amounts of 11D5 were higher in green tea than in black tea. Moreover, Darjeeling tea infusions were by ~30% richer in 11D5 than black and green teas from other regions. Each cup of green and black tea infusion may provide 20–60 μg 11D5, which is about 5% of the amounts found in tea samples. Spread over the day, regular tea consumption may contribute to the intake of valuable FuFAs.     

Co-Oxidative Transformation of Piperine to Piperonal and 3,4-Methylenedioxycinnamaldehyde by a Lipoxygenase from Pleurotus sapidus

The valuable aroma compound piperonal with its vanilla-like olfactory properties is of high interest for the fragrance and flavor industry. A lipoxygenase (LOX_Psa1) of the basidiomycete Pleurotus sapidus was identified to convert piperine, the abundant pungent principle of black pepper (Piper nigrum), to piperonal and a second volatile product, 3,4-methylenedioxycinnamaldehyde, with a vanilla-like odor through an alkene cleavage. The reaction principle was co-oxidation, as proven by its dependence on the presence of linoleic or α-linolenic acid, common substrates of lipoxygenases. Optimization of the reaction conditions (substrate concentrations, reaction temperature and time) led to a 24-fold and 15-fold increase of the piperonal and 3,4-methylenedioxycinnamaldehyde concentration using the recombinant enzyme. Monokaryotic strains showed different concentrations of and ratios between the two reaction products.     

A Versatile Route to Assemble Semiconductor Nanoparticles into Functional Aerogels by Means of Trivalent Cations

3D nanoparticle assemblies offer a unique platform to enhance and extend the functionality and optical/electrical properties of individual nanoparticles. Especially, a self‐supported, voluminous, and porous macroscopic material built up from interconnected semiconductor nanoparticles provides new possibilities in the field of sensing, optoelectronics, and photovoltaics. Herein, a method is demonstrated for assembling semiconductor nanoparticle systems containing building blocks possessing different composition, size, shape, and surface ligands. The method is based on the controlled destabilization of the particles triggered by trivalent cations (Y³⁺, Yb³⁺, and Al³⁺). The effect of the cations is investigated via X‐ray photoelectron spectroscopy. The macroscopic, self‐supported aerogels consist of the hyperbranched network of interconnected CdSe/CdS dot‐in‐rods, or CdSe/CdS as well as CdSe/CdTe core‐crown nanoplatelets is used to demonstrate the versatility of the procedure. The non‐oxidative assembly method takes place at room temperature without thermal activation in several hours and preserves the shape and the fluorescence of the building blocks. The assembled nanoparticle network provides longer exciton lifetimes with retained photoluminescence quantum yields, that make these nanostructured materials a perfect platform for novel multifunctional 3D networks in sensing. Various sets of photoelectrochemical measurements on the interconnected semiconductor nanorod structures also reveal the enhanced charge carrier separation.     

Patterning of Nanoparticle‐Based Aerogels and Xerogels by Inkjet Printing

Nanoparticle‐based voluminous 3D networks with low densities are a unique class of materials and are commonly known as aerogels. Due to the high surface‐to‐volume ratio, aerogels and xerogels might be suitable materials for applications in different fields, e.g. photocatalysis, catalysis, or sensing. One major difficulty in the handling of nanoparticle‐based aerogels and xerogels is the defined patterning of these structures on different substrates and surfaces. The automated manufacturing of nanoparticle‐based aerogel‐ or xerogel‐coated electrodes can easily be realized via inkjet printing. The main focus of this work is the implementation of the standard nanoparticle‐based gelation process in a commercial inkjet printing system. By simultaneously printing semiconductor nanoparticles and a destabilization agent, a 3D network on a conducting and transparent surface is obtained. First spectro‐electrochemical measurements are recorded to investigate the charge–carrier mobility within these 3D semiconductor‐based xerogel networks.     

Insights into the In Vitro Formation of Apatite from Mg-Stabilized Amorphous Calcium Carbonate

A protein-free formation of bone-like apatite from amorphous precursors through ball-milling is reported. Mg²⁺ ions are crucial to achieve full amorphization of CaCO₃. Mg²⁺ incorporation generates defects which strongly retard a recrystallization of ball-milled Mg-doped amorphous calcium carbonate (BM-aMCC), which promotes the growth of osteoblastic and endothelial cells in simulated body fluid and has no effect on endothelial cell gene expression. Ex situ snapshots of the processes revealed the reaction mechanisms. For low Mg contents (<30%) a two phase system consisting of Mg-doped amorphous calcium carbonate (ACC) and calcite “impurities” was formed. For high (>40%) Mg²⁺ contents, BM-aMCC follows a different crystallization path via magnesian calcite and monohydrocalcite to aragonite. While pure ACC crystallizes rapidly to calcite in aqueous media, Mg-doped ACC forms in the presence of phosphate ions bone-like hydroxycarbonate apatite (dahllite), a carbonate apatite with carbonate substitution in both type A (OH⁻) and type B (PO₄³⁻) sites, which grows on calcite “impurities” via heterogeneous nucleation. This process produces an endotoxin-free material and makes BM-aMCC an excellent “ion storage buffer” that promotes cell growth by stimulating cell viability and metabolism with promising applications in the treatment of bone defects and bone degenerative diseases.     

Metal–Ligand Based Mechanophores Enhance Both Mechanical Robustness and Electronic Performance of Polymer Semiconductors

The backbone of diketopyrrolopyrrole-thiophene-vinylene-thiophene-based polymer semiconductors (PSCs) is modified with pyridine (Py) or bipyridine ligands to complex Fe(II) metal centers, allowing the metal–ligand complexes to act as mechanophores and dynamically crosslink the polymer chains. Mono- and bi-dentate ligands are observed to exhibit different degrees of bond strengths, which subsequently affect the mechanical properties of these Wolf-type-II metallopolymers. The counter ion also plays a crucial role, as it is observed that Py-Fe mechanophores with non-coordinating BPh₄^– counter ions (Py-FeB) exhibit better thin film ductility with lower elastic modulus, as compared to the coordinating chloro ligands (Py-FeC). Interestingly, besides mechanical robustness, the electrical charge carrier mobility can also be enhanced concurrently when incorporating Py-FeB mechanophores in PSCs. This is a unique observation among stretchable PSCs, especially that most reports to date describe a decreased mobility when the stretchability is improved. Next, it is determined that improvements to both mobility and stretchability are correlated to the solid-state molecular ordering and dynamics of coordination bonds under strain, as elucidated via techniques of grazing-incidence X-ray diffraction and X-ray absorption spectroscopy techniques, respectively. This study provides a viable approach to enhance both the mechanical and the electronic performance of polymer-based soft devices.     

DNA Primer Extension with Cyclopropenylated 7‐Deaza‐2′‐deoxyadenosine and Efficient Bioorthogonal Labeling in Vitro and in Living Cells

A deoxyadenosine triphosphate (dATP) analogue for DNA labeling was synthesized with the 1‐methylcyclopropene (1MCP) group at the 7‐position of 7‐deaza‐2′‐deoxyadenosine and applied for primer extension experiments. The real‐time kinetic data reveals that this 1MCP‐modified dATP analogue is incorporated into DNA much faster than that of the similarly 1MCP‐modified deoxyuridine triphosphate (dUTP) analogue. The postsynthetic fluorescent labeling of these oligonucleotides works efficiently according to PAGE analysis, and can be applied for immobilization of a functional antibody on a surface. Site‐specific labeling at two different positions in DNA was achieved and the bioorthogonality of the postsynthetic fluorescent labeling was demonstrated in living HeLa cells.