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The potential of plant bioactives for the prevention and therapy of diabetes is increasingly being recognized. In the present study we investigated the antidiabetic properties of an aqueous Bistorta officinalis Delarbre extract (BODE) by employing both in-vitro assays and in-vivo models. Multiple targets in glucose homeostasis which are involved in the regulation of the blood glucose level were affected by BODE in-vitro. The extract exhibited inhibitory activities towards the intestinal carbohydrate-hydrolysing enzymes a-amylase and a-glucosidase with IC50 values of 81.5 µg/mL and 8.4 µg/mL, respectively. Furthermore, moderate reduction of the dipeptidyl peptidase-4 (DPP4) enzyme activity was evident when tested in the presence of 1.0 mg/mL BODE. A significant inhibition of the intestinal glucose transporter sodiumdependent glucose transporter 1 (SGLT1) in response to 1.0 mg/mL BODE was shown for Caco-2 cells mounted in Ussing chambers. High performance liquid chromatography-mass spectrometry analyses of the BODE revealed several plant bioactives including gallotannins, catechins and chlorogenic acid. Although our in-vitro data were promising, BODE-supplementation in the model organism Drosophila melanogaster lacked to confirm the antidiabetic effect of the extract in-vivo. Moreover, BODE failed to reduce blood glucose levels in chicken embryos (in-ovo). Hence, BODE is probably not a suitable candidate for developing a pharmaceutical against diabetes mellitus. [ABSTRACT FROM AUTHOR]

Etherified melamine resins are well known from the lacquer and paint industry. Up to now, important characterization in terms of composition and structure has been a time-consuming procedure. Now, fast analysis can also be achieved by use of 13C nuclear magnetic resonance spectroscopy. A special polarization transfer technique was applied to overcome the inherent low signal/noise ratio of 13C NMR spectra. The melamine content of the resin was determined by combustion analysis. This combination of methods allows for fast resin analysis with accuracy as yet unmatched and shows good correlation with established techniques. [ABSTRACT FROM AUTHOR]

Butyl octyl magnesium solutions are important raw materials in various chemical processes but suffer from their high reactivity with even traces of water, protic solvents or oxygen and an increased viscosity in hydrocarbon solution due to the formation of polymeric structures. N 1 ,N 2 -diphenylacenaphthylene-1,2-diimines (BIANs) have already been identified as potential candidates to reduce the viscosity of alkyl magnesium solutions and this study provides a systematic insight into the dependence of this ability on the position and structures of substituents on the BIAN. Besides the various BIANs, ZnCl 2 complexes and hydrogenated derivatives were characterized and tested for their ability to reduce the viscosity. HPLC-high resolution mass spectrometry, MALDI-ToF mass spectrometry, but most important FTIR and NMR experiments under inert conditions have been used to shine light on the interaction of the different BIAN derivatives with alkyl magnesium solutions. Hydrogenated BIANs, especially those with bulky alkyl groups in the ortho position(s) have been identified as the most promising candidates. An additional benefit of the hydrogenated species is that in contrast to BIANs and BIAN-Zn complexes they do not undergo permanent chemical modification and can be reused after extraction., (© 2024. The Author(s).)

Diabetes mellitus is a spreading global pandemic. Type 2 diabetes mellitus (T2DM) is the predominant form of diabetes, in which a reduction in blood glucose uptake is caused by impaired glucose transporter 4 (GLUT4) translocation to the plasma membrane in adipose and muscle cells. Antihyperglycemic drugs play a pivotal role in ameliorating diabetes symptoms but often are associated with side effects. Hence, novel antidiabetic compounds and nutraceutical candidates are urgently needed. Phytogenic therapy can support the prevention and amelioration of impaired glucose homeostasis. Using total internal reflection fluorescence microscopy (TIRFM), 772 plant extracts of an open-access plant extract library were screened for their GLUT4 translocation activation potential, resulting in 9% positive hits. Based on commercial interest and TIRFM assay-based GLUT4 translocation activation, some of these extracts were selected, and their blood glucose-reducing effects in ovo were investigated using a modified hen's egg test (Gluc-HET). To identify the active plant part, some of the available candidate plants were prepared in-house from blossoms, leaves, stems, or roots and tested. Acacia catechu (catechu), Pulmonaria officinalis (lungwort), Mentha spicata (spearmint), and Saponaria officinalis (common soapwort) revealed their potentials as antidiabetic nutraceuticals, with common soapwort containing GLUT4 translocation-activating saponarin.

Four pinaceae pine resins analyzed in this study: black pine, shore pine, Baltic amber, and rosin demonstrate excellent dielectric properties, outstanding film forming, and ease of processability from ethyl alcohol solutions. Their trap-free nature allows fabrication of virtually hysteresis-free organic field effect transistors operating in a low voltage window with excellent stability under bias stress. Such green constituents represent an excellent choice of materials for applications targeting biocompatibility and biodegradability of electronics and sensors, within the overall effort of sustainable electronics development and environmental friendliness., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors. Global Challenges published by Wiley‐VCH GmbH.)

Wearable sweat sensors can potentially be used to continuously and non-invasively monitor physicochemical biomarkers that contain information related to disease diagnostics and fitness tracking. However, the development of such autonomous sensors faces a number of challenges including achieving steady sweat extraction for continuous and prolonged monitoring, and addressing the high power demands of multifunctional and complex analysis. Here we report an autonomous wearable biosensor that is powered by a perovskite solar cell and can provide continuous and non-invasive metabolic monitoring. The device uses a flexible quasi-two-dimensional perovskite solar cell module that provides ample power under outdoor and indoor illumination conditions (power conversion efficiency exceeding 31% under indoor light illumination). We show that the wearable device can continuously collect multimodal physicochemical data - glucose, pH, sodium ions, sweat rate, and skin temperature - across indoor and outdoor physical activities for over 12 hours., Competing Interests: Competing interests The authors declare no competing interests.

The newly developed MALDI interpretation tool (" MALINTO ") allows for the accelerated characterization of complex synthetic polymers via MALDI mass spectrometry. While existing software provides solutions for simple polymers like poly(ethylene glycol), polystyrene, etc., they are limited in their application on polycondensates synthesized from two different kinds of monomers (e.g., diacid and diol in polyesters). In addition to such A 2 + B 2 polycondensates, MALINTO covers branched and even multicyclic polymer systems. Since the MALINTO software works based on input data of monomers/repeating units, end groups, and adducts, it can be applied on polymers whose components are previously known or elucidated. Using these input data, a list with theoretically possible polymer compositions and resulting m / z values is calculated, which is further compared to experimental mass spectrometry data. For optional semiquantitative studies, peak areas are allocated according to their assigned polymer composition to evaluate both comonomer and terminating group ratios. Several tools are implemented to avoid mistakes, for example, during peak assignment. In the present publication, the functions of MALINTO are described in detail and its broad applicability on different linear polymers as well as branched and multicyclic polycondensates is demonstrated. Fellow researchers will benefit from the accelerated peak assignment using the freely available MALINTO software and might be encouraged to explore the potential of MALDI mass spectrometry for (semi)quantitative applications.

N 1 -diphenylacenaphthylene-1,2-diimines (BIANs) have been used to reduce the undesired high viscosity of alkyl magnesium solutions, which are known to form polymeric structures. In order to understand the mechanisms, analyses of the BIAN alkyl magnesium solutions have been carried out under inert conditions with SEC-MS, NMR, and FTIR and were compared to the structures obtained from HPLC-MS, FTIR, and NMR after aqueous workup. While viscosity reduction was shown for all BIAN derivatives used, only the bis (diisopropyl)-substituted BIAN could be clearly assigned to a single reaction product, which also could be reused without loss of efficiency or decomposition. All other derivatives have been shown to behave differently, even under inert conditions, and decompose upon contact with acidic solvents. While the chemical reactions observed after the workup of the used BIANs are dominated by (multiple) alkylation, mainly on the C = N double bond, the observation of viscosity reduction cannot be assigned to this reaction alone, but to the interaction of the nitrogen atoms of BIANs with the Mg of the alkyl magnesium polymers, as could be shown by FTIR and NMR measurements under inert conditions.N 2 -diphenylacenaphthylene-1,2-diimines (BIANs) have been used to reduce the undesired high viscosity of alkyl magnesium solutions, which are known to form polymeric structures. In order to understand the mechanisms, analyses of the BIAN alkyl magnesium solutions have been carried out under inert conditions with SEC-MS, NMR, and FTIR and were compared to the structures obtained from HPLC-MS, FTIR, and NMR after aqueous workup. While viscosity reduction was shown for all BIAN derivatives used, only the bis (diisopropyl)-substituted BIAN could be clearly assigned to a single reaction product, which also could be reused without loss of efficiency or decomposition. All other derivatives have been shown to behave differently, even under inert conditions, and decompose upon contact with acidic solvents. While the chemical reactions observed after the workup of the used BIANs are dominated by (multiple) alkylation, mainly on the C = N double bond, the observation of viscosity reduction cannot be assigned to this reaction alone, but to the interaction of the nitrogen atoms of BIANs with the Mg of the alkyl magnesium polymers, as could be shown by FTIR and NMR measurements under inert conditions.

Scope: Sea buckthorn (Hippophaes rhamnoides) is capable of ameliorating disturbed glucose metabolism in animal models and human subjects. Here, the effect of sea buckthorn oil as well as of extracts of fruits, leaves, and press cake on postprandial glucose metabolism is systematically investigated., Methods and Results: Sea buckthorn did neither exert decisive effects in an in vitro model of intestinal glucose absorption nor did it alter insulin secretion. However, sea buckthorn stimulates GLUT4 translocation to the plasma membrane comparable to insulin, indicative of increased glucose clearance from the circulation. Isorhamnetin is identified in all sea buckthorn samples investigated and is biologically active in triggering GLUT4 cell surface localization. Consistently, sea buckthorn products lower circulating glucose by ≈10% in a chick embryo model. Moreover, sea buckthorn products fully revert hyperglycemia in the nematode Caenorhabditis elegans while they are ineffective in Drosophila melanogaster under euglycemic conditions., Conclusion: These data indicate that edible sea buckthorn products as well as by-products are promising resources for hypoglycemic nutrient supplements that increase cellular glucose clearance into target tissues., (© 2022 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH GmbH.)

Medicinal plant extracts are becoming increasingly important as an alternative for traditional drugs against diabetes mellitus (DM). For this reason, we initialized a target-based screening of 111 root extracts from an open access plant extract library (PECKISH) by ascertaining their in-vitro inhibitory efficacy on α-glucosidase. The two most active extracts Geum urbanum L. (roseroot) and Rhodiola rosea L. (avens root) were further tested for their antidiabetic activities in terms of their impact on different regulatory key points of glucose homeostasis. To this end, various enzyme- and cell culture-based in-vitro assays were employed including the determination of sodium-dependent glucose transporter 1 (SGLT1) activity in Caco-2 monolayers by Ussing chambers and of glucose transporter 4 (GLUT4) translocation in a GFP-reporter cell line. Subsequently, the antidiabetic potential of the root extracts were further evaluated in in-vivo models, namely hen's eggs test and the fruit fly Drosophila melanogaster . Avens root extract was found to be a more potent inhibitor of the enzymes α-glucosidase and dipeptidyl peptidase-4 (DPP4) than roseroot extract. Most importantly, only avens root extract exhibited antidiabetic activity in the two in-vivo models eliciting a reduced blood glucose level in the in-ovo model and a decline of the triglyceride level in a dietary starch-induced D. melanogaster obesity model. Analyses of the polyphenolic composition of the avens root extract by HPLC revealed a high content of ellagic acid and its derivatives as well as ellagitannins such as pedunculagin, stenophyllanin, stachyurin, casuarinin and gemin A. In conclusion, avens root extract represents a promising medicinal plant that should be considered in further in-vivo studies on hyperglycemia in laboratory rodents and humans., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Günther, Rimbach, Nevermann, Neuhauser, Stadlbauer, Schwarzinger, Schwarzinger, Ipharraguerre, Weghuber and Lüersen.)

Decreasing circulating low-density lipoprotein (LDL) cholesterol levels leads to decreased risk of cardiovascular diseases. Natural compounds are capable of lowering LDL-cholesterol even on top of lifestyle modification or medication. To identify novel plant-derived compounds to lower plasma LDL cholesterol levels, we performed high-content screening based on the transcriptional activation of the promoter of the LDL receptor (LDLR). The identified hits were thoroughly validated in human hepatic cell lines in terms of increasing LDLR mRNA and protein levels, lowering cellular cholesterol levels and increasing cellular LDL uptake. By means of this incremental validation process in vitro , aqueous extracts prepared from leaves of lingonberries ( Vaccinium vitis-idaea ) as well as blackberries ( Rubus fruticosus ) were found to have effects comparable to lovastatin, a prototypic cholesterol-lowering drug. When applied in vivo in mice, both extracts induced subtle increases in hepatic LDLR expression. In addition, a significant increase in high-density lipoprotein (HDL) cholesterol was observed. Taken together, aqueous extracts from lingonberry or blackberry leaves were identified and characterized as strong candidates to provide cardiovascular protection.

Matrix-assisted laser desorption ionization/time-of-flight (MALDI/ToF) mass spectrometry and 1 H NMR were used for the structural investigation of isophthalic and maleic acid copolyesters with neopentyl glycol. Since both methods provided information on the ratio of incorporated acid components and terminating groups, results were compared and linear correlations ( R 2 = 0.96-0.98) could be found. This suggests that MALDI/ToF MS is a suitable tool for the semiquantitative characterization of polyester systems. For the isophthalic/maleic acid ratio, MALDI results yielded constantly lower values than 1 H NMR, which was attributed to varying ionization efficiencies of homo- and copolyesters. Ratios of carboxylic and hydroxylic terminating groups, which are conventionally still measured by time consuming complex titrations, were measured with MALDI and 1 H NMR and were in good agreement. Both methods either excluded or distinguished unreacted monomers in the polyester bulk in contrast to acid-base titrations where those monomers severely distort the results. Additional structural information could be gained including the observation of cyclic structures (MALDI), E / Z isomerism from maleic to fumaric acid, and the statistical distribution of the acid components within the polyester chain ( 1 H NMR). While 1 C NMR and multidimensional techniques, MALDI/ToF MS provides a straightforward technique that can be applied to other polyester systems without major alterations.13 C NMR and multidimensional techniques, MALDI/ToF MS provides a straightforward technique that can be applied to other polyester systems without major alterations.

Climatic changes and heat stress have become a great challenge in the livestock industry, negatively affecting, in particular, poultry feed intake and intestinal barrier malfunction. Recently, phytogenic feed additives were applied to reduce heat stress effects on animal farming. Here, we investigated the effects of ginseng extract using various in vitro and in vivo experiments. Quantitative real-time PCR, transepithelial electrical resistance measurements and survival assays under heat stress conditions were carried out in various model systems, including Caco-2 cells, Caenorhabditis elegans and jejunum samples of broilers. Under heat stress conditions, ginseng treatment lowered the expression of HSPA1A (Caco-2) and the heat shock protein genes hsp-1 and hsp-16.2 (both in C. elegans ), while all three of the tested genes encoding tight junction proteins, CLDN3 , OCLN and CLDN1 (Caco-2), were upregulated. In addition, we observed prolonged survival under heat stress in Caenorhabditis elegan s , and a better performance of growing ginseng-fed broilers by the increased gene expression of selected heat shock and tight junction proteins. The presence of ginseng extract resulted in a reduced decrease in transepithelial resistance under heat shock conditions. Finally, LC-MS analysis was performed to quantitate the most prominent ginsenosides in the extract used for this study, being Re, Rg1, Rc, Rb2 and Rd. In conclusion, ginseng extract was found to be a suitable feed additive in animal nutrition to reduce the negative physiological effects caused by heat stress., Competing Interests: The authors declare no conflict of interest.

The transport of hydrophobic compounds to recipient cells is a critical step in nutrient supplementation. Here, we tested the effect of phospholipid-based emulsification on the uptake of hydrophobic compounds into various tissue culture cell lines. In particular, the uptake of ω-3 fatty acids from micellar or nonmicellar algae oil into cell models for enterocytes, epithelial cells, and adipocytes was tested. Micellization of algae oil did not result in adverse effects on cell viability in the target cells. In general, both micellar and nonmicellar oil increased intracellular docosahexaenoic acid (DHA) levels. However, micellar oil was more effective in terms of augmenting the intracellular levels of total polyunsaturated fatty acids (PUFAs) than nonmicellar oil. These effects were rather conserved throughout the cells tested, indicating that fatty acids from micellar oils are enriched by mechanisms independent of lipases or lipid transporters. Importantly, the positive effect of emulsification was not restricted to the uptake of fatty acids. Instead, the uptake of phytosterols from phytogenic oils into target cells also increased after micellization. Taken together, phospholipid-based emulsification is a straightforward, effective, and safe approach to delivering hydrophobic nutrients, such as fatty acids or phytosterols, to a variety of cell types in vitro. It is proposed that this method of emulsification is suitable for the effective supplementation of numerous hydrophobic nutrients., Competing Interests: The authors declare no conflicts of interest. PM International AG provided support in the form of the salaries for M.I. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

An increase in adipose tissue is caused by the increased size and number of adipocytes. Lipids accumulate in intracellular stores, known as lipid droplets (LDs). Recent studies suggest that parameters such as LD size, shape and dynamics are closely related to the development of obesity. Berberine (BBR), a natural plant alkaloid, has been demonstrated to possess anti-obesity effects. However, it remains unknown which cellular processes are affected by this compound or how effective herbal extracts containing BBR and other alkaloids actually are. For this study, we used extracts of Coptis chinensis, Mahonia aquifolium, Berberis vulgaris and Chelidonium majus containing BBR and other alkaloids and studied various processes related to adipocyte functionality. The presence of extracts resulted in reduced adipocyte differentiation, as well as neutral lipid content and rate of lipolysis. We observed that the intracellular fatty acid exchange was reduced in different LD size fractions upon treatment with BBR and Coptis chinensis. In addition, LD motility was decreased upon incubation with BBR, Coptis chinensis and Chelidonium majus extracts. Furthermore, Chelidonium majus was identified as a potent fatty acid uptake inhibitor. This is the first study that demonstrates the selected regulatory effects of herbal extracts on adipocyte function.

Inhibition of intestinal glucose resorption can serve as an effective strategy for the prevention of an increase in blood glucose levels. We have recently shown that various extracts prepared from guava ( Psidium guajava ) inhibit sodium-dependent glucose cotransporter 1 (SGLT1)- and glucose transporter 2 (GLUT2)-mediated glucose transport in vitro (Caco-2 cells) and in vivo (C57BL/6N mice). However, the efficacy in humans remains to be confirmed. For this purpose, we conducted a parallelized, randomized clinical study with young healthy adults. Thirty-one volunteers performed an oral glucose tolerance test (OGTT) in which the control group received a glucose solution and the intervention group received a glucose solution containing a guava fruit extract prepared by supercritical CO 2 extraction. The exact same extract was used for our previous in vitro and in vivo experiments. Blood samples were collected prior to and up to two hours after glucose consumption to quantitate blood glucose and insulin levels. Our results show that, in comparison to the control group, consumption of guava fruit extract resulted in a significantly reduced increase in postprandial glucose response over the basal fasting plasma glucose levels after 30 min (Δ control 2.60 ± 1.09 mmol/L versus Δ intervention 1.96 ± 0.96 mmol/L; p = 0.039) and 90 min (Δ control 0.44 ± 0.74 mmol/L versus Δ intervention -0.18 ± 0.88 mmol/L; p = 0.023). In addition, we observed a slightly reduced, but non-significant insulin secretion (Δ control 353.82 ± 183.31 pmol/L versus Δ intervention 288.43 ± 126.19 pmol/L, p = 0.302). Interestingly, storage time and repeated freeze-thawing operations appeared to negatively influence the efficacy of the applied extract. Several analytical methods (HPLC-MS, GC-MS, and NMR) were applied to identify putative bioactive compounds in the CO 2 extract used. We could assign several substances at relevant concentrations including kojic acid (0.33 mg/mL) and 5-hydroxymethylfurfural (2.76 mg/mL). Taken together, this clinical trial and previous in vitro and in vivo experiments confirm the efficacy of our guava fruit extract in inhibiting intestinal glucose resorption, possibly in combination with reduced insulin secretion. Based on these findings, the development of food supplements or functional foods containing this extract appears promising for patients with diabetes and for the prevention of insulin resistance. Trial registration: 415-E/2319/15-2018 (Ethics Commissions of Salzburg).

2-Nitrobenzaldehyde was found to efficiently block singlet oxygen generation in a series of different test samples upon exposure to UV and visible light under aerobic conditions. The effect of quenching singlet oxygen formation was monitored in the presence of 1, 4-diazabicyclo [2.2.2] octane (DABCO) acting as a well-known singlet oxygen scavenger. A comparison of different nitrobenzaldehyde isomers with other highly effective synthetic antioxidants used in the food industry such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tert-butylhydroquinone (TBHQ) revealed that the protection of materials from singlet oxygen decreases in the order of 2-nitrobenzaldehyde > DABCO > TBHQ > 3-nitrobenzaldehyde > BHA > 4-nitrobenzaldehyde > BHT. Upon addition of 2-nitrobenzaldehyde, the oxidation of fatty acids and the degradation of photosensitizers was found to be considerably diminished, which indicates that the presence of 2-nitrobenzaldehyde has a significant protective influence by restricting the singlet oxygen generation and photodegradation of dyes. Moreover, the compound turned out to display its highly suppressing effects on typical singlet oxygen-dependent reactions, such as fatty acid photooxidation and dye photosensitizer degradation, in a rather broad spectral region covering wavelengths from 300 nm (UV-B) to 575 nm (close to the maximum of ambient solar radiation).

Scope: Known pharmacological activities of guava (Psidium guajava) include modulation of blood glucose levels. However, mechanistic details remain unclear in many cases., Methods and Results: This study investigated the effects of different guava leaf and fruit extracts on intestinal glucose transport in vitro and on postprandial glucose levels in vivo. Substantial dose- and time-dependent glucose transport inhibition (up to 80%) was observed for both guava fruit and leaf extracts, at conceivable physiological concentrations in Caco-2 cells. Using sodium-containing (both glucose transporters, sodium-dependent glucose transporter 1 [SGLT1] and glucose transporter 2 [GLUT2], are active) and sodium-free (only GLUT2 is active) conditions, we show that inhibition of GLUT2 was greater than that of SGLT1. Inhibitory properties of guava extracts also remained stable after digestive juice treatment, indicating a good chemical stability of the active substances. Furthermore, we could unequivocally show that guava extracts significantly reduced blood glucose levels (≈fourfold reduction) in a time-dependent manner in vivo (C57BL/6N mice). Extracts were characterized with respect to their main putative bioactive compounds (polyphenols) using HPLC and LC-MS., Conclusion: The data demonstrated that guava leaf and fruit extracts can potentially contribute to the regulation of blood glucose levels., (© 2018 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

N-Methylmelamines have recently gained importance as valuable compounds for manufacturing modified melamine formaldehyde resins and other polymer building blocks. A great advantage of these polymers is the reduction of the carcinogenic formaldehyde. Selecting the polymerization processes (e.g., substance polymerization, polymerization in solution) and controlling the polymerization reaction and properties of these novel materials requires knowledge of the properties of the individual melamine derivatives used as new building blocks. All possible permutations of N-methylmelamines were prepared, and reaction progress was monitored by GC/MS. 2,4,6-Tris(dimethylamino)-1,3,5-triazine was prepared to complete the series; this is, however, also a possible byproduct in various synthesis routes. The reaction conditions were optimized to obtain high yields of each derivative with the highest possible purity. The substances were characterized by NMR and IR spectroscopy, mass spectrometry, elemental analysis, and single-crystal X-ray diffraction. In addition, physical properties, such as solubility, melting points, and pKb values, were determined. The number of amino-, methylamino-, and dimethylamino groups has a significant effect on these properties. In summary, we found that by increasing the number of amino- and methylamino groups, solubility and pKb increase. With increasing number of amino groups, the compounds tend to form hydrogen bonds, and thus, the melting point shifts to higher temperature ranges where they start to decompose.

The segregation in dried droplet MALDI sample spots was analyzed with regard to the matrix-to-sample ratio using optical microscopy, MALDI imaging mass spectrometry (MALDI MSI) and IR imaging spectroscopy. In this context, different polymer/matrix/solvent systems usually applied in the analysis of synthetic polymers were investigated. The use of typical matrix concentrations (10 mg mL⁻¹) in almost every case resulted in ring patterns, whereas higher concentrated matrix solutions always led to homogeneous sample spot layers. The data revealed that segregation is predominantly caused by matrix transport in the drying droplet, whereas polymer segregation seems to be only secondary.

Rationale: Polymer sample spots for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) prepared by the dried-droplet method often reveal ring formation accompanied by possible segregation of matrix and sample molecules as well as of the polymer homologs itself. Since the majority of sample spots are prepared by this simple and fast method, a matrix or sample preparation method that excludes such segregation has to be found., Methods: Three different ionic liquid matrices based on conventionally used aromatic compounds for MALDI-TOF MS were prepared. The formation of ionic liquids was proven by (1) H NMR spectroscopy. MALDI-Imaging mass spectrometry was applied to monitor the homogeneity., Results: Our results show a superior sample spot homogeneity using ionic liquid matrices. Spots could be sampled several times without visible differences in the mass spectra. A frequently observed loss of matrix in the mass spectrometer vacuum was not observed. The necessary laser irradiance was reduced, which resulted in less polymer fragmentation., Conclusions: Ionic liquid matrices can be used to overcome segregation, a typical drawback of conventional MALDI dried-droplet preparations. Homogeneous sample spots are easy to prepare, stable in the MS vacuum and, thereby, improve the reproducibility of MALDI., (Copyright © 2014 John Wiley & Sons, Ltd.)

The first record of millipedes (Diplopoda) being regularly used for food by humans (the Bobo people of Burkina Faso) is given, including information on how the millipedes are prepared. The species in question are Tymbodesmus falcatus (Karsch, 1881) and Sphenodesmus sheribongensis (Schiøtz, 1966) (Gomphodesmidae) and an unidentified species of Spirostreptidae. New information on the nutritional value of millipedes is provided; unsaturated fatty acids, calcium, and iron contents are particularly high. The millipedes' defensive secretions, hydrogen cyanide and benzoquinones, present a severe challenge for the spread of millipedes as an everyday food source. On the other hand, the possibility that benzoquinones may act as insect-repellents, as known from studies on nonhuman primates, and that sublethal cyanide ingestion may enhance human innate resistance to malaria, suggests promising ethnomedical perspectives to our findings.

Label-free biosensors detect binding of prey molecules (″analytes″) to immobile bait molecules on the sensing surface. Numerous methods are available for immobilization of bait molecules. A convenient option is binding of biotinylated bait molecules to streptavidin-functionalized surfaces, or to biotinylated surfaces via biotin-avidin-biotin bridges. The goal of this study was to find a rapid method for reversible immobilization of biotinylated bait molecules on biotinylated sensor chips. The task was to establish a biotin-avidin-biotin bridge which was easily cleaved when desired, yet perfectly stable under a wide range of measurement conditions. The problem was solved with the avidin mutant M96H which contains extra histidine residues at the subunit-subunit interfaces. This mutant was bound to a mixed self-assembled monolayer (SAM) containing biotin residues on 20% of the oligo(ethylene glycol)-terminated SAM components. Various biotinylated bait molecules were bound on top of the immobilized avidin mutant. The biotin-avidin-biotin bridge was stable at pH ≥3, and it was insensitive to sodium dodecyl sulfate (SDS) at neutral pH. Only the combination of citric acid (2.5%, pH 2) and SDS (0.25%) caused instantaneous cleavage of the biotin-avidin-biotin bridge. As a consequence, the biotinylated bait molecules could be immobilized and removed as often as desired, the only limit being the time span for reproducible chip function when kept in buffer (2-3 weeks at 25 °C). As expected, the high isolectric pH (pI) of the avidin mutant caused nonspecific adsorption of proteins. This problem was solved by acetylation of avidin (to pI < 5), or by optimization of SAM formation and passivation with biotin-BSA and BSA.

The alkynyl-substituted Re I complex [Re(5,5'-bisphenylethynyl-2,2'-bipyridyl)(CO) 3 Cl] was immobilized by electropolymerization onto a Pt-plate electrode. The polymerized film exhibited electrocatalytic activity for the reduction of CO 2 to CO. Cyclic voltammetry studies and bulk controlled-potential electrolysis experiments were performed by using a CO 2 -saturated acetonitrile solution. The CO 2 reduction, determined by cyclic voltammetry, occurs at approximately -1150 mV versus the normal hydrogen electrode (NHE). Quantitative analysis by GC and IR spectroscopy was used to determine a Faradaic efficiency of approximately 33 % for the formation of CO. Both values of the modified electrode were compared to the performance of the homogenous monomer [Re(5,5'-bisphenylethynyl-2,2'-bipyridyl)(CO) 3 Cl] in acetonitrile. The polymer formation and its properties were studied by using SEM, AFM, and attenuated total reflectance (ATR) FTIR and UV/Vis spectroscopy.

MALDI-TOF MS is used for the qualitative analysis of seven different polymer additives directly from the polymer without tedious sample pretreatment. Additionally, by using a solid sample preparation technique, which avoids the concentration gradient problems known to occur with dried droplets and by adding tetraphenylporphyrine as an internal standard to the matrix, it is possible to perform quantitative analysis of additives directly from the polymer sample. Calibration curves for Tinuvin 770, Tinuvin 622, Irganox 1024, Irganox 1010, Irgafos 168, and Chimassorb 944 are presented, showing coefficients of determination between 0.911 and 0.990.

Random spin labeling (RSL) is a method for rapid mapping of biomolecular interaction surfaces using an interaction partner with SL and an interaction partner enriched in (13)C or (15)N nuclei for paramagnetic relaxation enhanced NMR-based detection. The SL reaction is conducted in a manner resulting in a heterogeneous reaction product consisting of different populations of the protein carrying a varying number of spin labels at different positions. Preparation of the paramagnetic probe is complete within a few hours and hence much faster than site selective SL. RSL is applicable to tightly interacting systems but shows its particular strength when applied to systems involving weak or transient contacts.

Background: Knowing the underlying mechanisms of mosquito ecology will ensure effective vector management and contribute to the overall goal of malaria control. Mosquito populations show a high degree of population plasticity in response to environmental variability. However, the principle factors controlling population size and fecundity are for the most part unknown. Larval habitat and diet play a crucial role in subsequent mosquito fitness. Developing the most competitive insects for sterile insect technique programmes requires a "production" orientated perspective, to deduce the most effective larval diet formulation; the information gained from this process offers us some insight into the mechanisms and processes taking place in natural native mosquito habitats., Methodology/principal Findings: Fatty acid profiles and de-novo or direct assimilation pathways, of whole-individual mosquitoes reared on a range of larval diets were determined using pyrolysis gas chromatograph/mass spectrometry. We used elemental analysis and isotope ratio mass spectrometry to measure individual-whole-body carbon, nitrogen and phosphorous values and to assess the impact of dietary quality on subsequent population stoichiometry, size, quality and isotopic signature. Diet had the greatest impact on fatty acid (FA) profiles of the mosquitoes, which exhibited a high degree of dietary routing, characteristic of generalist feeders. De-novo synthesis of a number of important FAs was observed. Mosquito C:N stoichiometry was fixed in the teneral stage. Dietary N content had significant influence on mosquito size, and P was shown to be a flexible pool which limited overall population size., Conclusions/significance: Direct routing of FAs was evident but there was ubiquitous de-novo synthesis suggesting mosquito larvae are competent generalist feeders capable of survival on diet with varying characteristics. It was concluded that nitrogen availability in the larval diet controlled teneral mosquito size and that teneral CN ratio is a sex- and species-specific fixed parameter. This finding has significant implications for overall mosquito competitiveness and environmental management.

STIM1 and Orai represent the key components of Ca(2+) release-activated Ca(2+) channels. Activation of Orai channels requires coupling of the C terminus of STIM1 to the N and C termini of Orai. Although the latter appears to be central in the interaction with STIM1, the role of the N terminus and particularly of the conserved region close to the first transmembrane sequence is less well understood. Here, we investigated in detail the functional role of this conserved region in Orai3 by stepwise deletions. Molecular determinants were mapped for the two modes of Orai3 activation via STIM1 or 2-aminoethoxydiphenyl borate (2-APB) and for current gating characteristics. Increasing N-terminal truncations revealed a progressive decrease of the specific fast inactivation of Orai3 concomitant with diminished binding to calmodulin. STIM1-dependent activation of Orai3 was maintained as long as the second half of this conserved N-terminal domain was present. Further truncations abolished it, whereas Orai3 stimulation via 2-APB was partially retained. In aggregate, the N-terminal conserved region plays a multifaceted role in Orai3 current gating with distinct structural requirements for STIM1- and 2-APB-stimulated activation.

A fast and reliable method based on CZE coupled to Q-TOF MS for the analysis of methylated melamines is presented. Experiments with UV detection were performed using BGE containing formic acid and TFA to optimize electrophoretic conditions and to achieve baseline separation. Employing a TFA-based electrolyte containing 80% ACN and MS detection, nine different methyl melamines were determined in real samples, which were produced by catalytic hydrogenation of melamine/formaldehyde reaction mixtures. The use of a Q-TOF MS resulted in LOD better than 0.01 mg/L for all compounds and high mass accuracy with mass errors lower than 2.3 ppm.

Biomineralization research on mollusc shells has mostly focused on nacre formation. Chitin, silk fibroin protein, and acidic macromolecules are important components for shell formation. Although the principle concept behind shell calcification was developed many years ago, the individual components have not been well scrutinized. Besides that, Mollusca are the second largest invertebrate phylum, but comprehensive biochemical research involving a comparison of different taxa is still rare. This study reconsiders the above three components with adding some biochemical data of aculiferans. The presence of chitin in polyplacophorans sclerites was confirmed by IR and pyrolysis GC/MS. DMMB staining data inferred that sulphated groups present in aplacophoran cuticle but not in polyplacophorans cuticle. These insight suggested importance of comparison between acuriferans and conchiferans.

Chitin is an insoluble component in the shells of several molluscan species. It is thought to play important roles, in biomineralization and shell structure. To date, however, reports are scarce and sometimes contradictory, and suffer from methodological problems. Only in a single cephalopod species has the chitin been identified as beta-chitin. We present data on chitin occurrence in 22 species of shell-bearing Mollusca (Conchifera) and Polyplacophora, including the first evidence for scaphopods, based on pyrolysis gas chromatography, mass spectrometry (GC-MS), and infrared spectroscopy (IR). Pyrolysis GC-MS detected chitin in every tested member of the Conchifera. IR spectroscopy before and after chitinase treatment revealed at least three distinct patterns of peak changes. The contents of the insoluble shell organics included not only chitin and proteins, but also insoluble polysaccharides, e.g., glucan. We conclude that chitin was present in the last common ancestor of the Conchifera and that its abundance in the shell matrix depends on the differentiation of the shell.

An improved method based on capillary zone electrophoresis (CZE) coupled to either ion-trap (IT) mass spectrometry (MS) or quadrupole-time-of-flight (Q-TOF) MS for the analysis of melamine-formaldehyde condensates is presented. Employing a formic acid-based electrolyte containing 50% acetonitrile (ACN) and MS detection up to 13 compounds could be determined in lab-made resins, synthesized by mixing formaldehyde and melamine in different ratios ranging from 1:1.5 to 1:4. The use of a Q-TOF-MS for detection allowed the assignment of molecular formulas for all 13 substances with high accuracy.

Flavonoid content and composition in 124 accessions of forage and grain legumes was assessed by means of HPLC-DAD and HPLC-ESI-MS analyses of seed extracts. Excluding soybean, only 7 out of 77 grain legume accessions exhibited a total flavonoid content higher than 0.1 mg/g fresh seed weight (mean content 0.33 +/- 0.31 mg/ g fresh seed weight); while in 38 out of 47 forage legume, the mean flavonoid content was 1.49 +/- 0.89 mg/g fresh seed weight. On the basis of their higher seed flavonoid content and agronomic relevance, two grain and five forage legume accessions were selected for further HPLC-ESI-MS analyses aimed to identify bioactive compounds. Chemical analysis of the selected accessions permitted the identification of several flavonoids with pharmaceutical properties such as vitexin in Vigna angularis and Trigonella foenum-graecum, quercetin in all Trifolium spp and Medicago sativa, taxifolin in Trifolium pratense and Trifolium alexandrinum, myricetin in Trifolium repens, kaempferol in Phaseolus vulgaris and T. repens and luteolin in M. sativa and T. alexandrinum. Since much of literature information is regarding flavonoid content and composition of the green part of legume species, the obtained results contributed to fill the gap in knowledge about flavonoid content at the seed level.