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Forensic DNA profiles are established by multiplex PCR amplification of a set of highly variable short tandem repeat (STR) loci followed by capillary electrophoresis (CE) as a means to assign alleles to PCR products of differential length. Recently, CE analysis of STR amplicons has been supplemented by high-throughput next generation sequencing (NGS) techniques that are able to detect isoalleles bearing sequence polymorphisms and allow for an improved analysis of degraded DNA. Several such assays have been commercialised and validated for forensic applications. However, these systems are cost-effective only when applied to high numbers of samples. We report here an alternative, cost-efficient shallow-sequence output NGS assay called maSTR assay that, in conjunction with a dedicated bioinformatics pipeline called SNiPSTR, can be implemented with standard NGS instrumentation. In a back-to-back comparison with a CE-based, commercial forensic STR kit, we find that for samples with low DNA content, with mixed DNA from different individuals, or containing PCR inhibitors, the maSTR assay performs equally well, and with degraded DNA is superior to CE-based analysis. Thus, the maSTR assay is a simple, robust and cost-efficient NGS-based STR typing method applicable for human identification in forensic and biomedical contexts.
Die Freiheit in Forschung und Lehre ist zusammen mit der Wissenschaftsfreiheit eines der bürgerlichen Grundrechte in Deutschland. Aber die Forschungsfreiheit kann nicht grenzenlos sein. Industrie wie akademische Institute müssen ihre Entwicklungen dokumentieren. Nur so ist ein Stoff später für die Vermarktung zulassungsfähig und nur so lässt sich belegen, wer die Urheberrechte daran hat.
Mutations in SELENBP1, encoding a novel human methanethiol oxidase, cause extraoral halitosis
(2017)
The application of Raman and infrared (IR) microspectroscopy is leading to hyperspectral data containing complementary information concerning the molecular composition of a sample. The classification of hyperspectral data from the individual spectroscopic approaches is already state-of-the-art in several fields of research. However, more complex structured samples and difficult measuring conditions might affect the accuracy of classification results negatively and could make a successful classification of the sample components challenging. This contribution presents a comprehensive comparison in supervised pixel classification of hyperspectral microscopic images, proving that a combined approach of Raman and IR microspectroscopy has a high potential to improve classification rates by a meaningful extension of the feature space. It shows that the complementary information in spatially co-registered hyperspectral images of polymer samples can be accessed using different feature extraction methods and, once fused on the feature-level, is in general more accurately classifiable in a pattern recognition task than the corresponding classification results for data derived from the individual spectroscopic approaches.
Durch Dotierung eines nematischen Flüssigkristalles mit einer chiralen Substanz wird eine helikal strukturierte Phase induziert, die in der Lage ist, einfallendes Licht wellenlängenselektiv zu reflektieren. Bei der Reaktion des Dotiermittels mit einem gasförmigen Analyten verändern sich die Ganghöhe dieser Struktur und damit die reflektierte Wellenlänge. Liegt diese im Bereich des sichtbaren Lichts, ist eine Farbänderung mit dem menschlichen Auge zu beobachten. Es ist dabei sinnvoll den Flüssigkristall z.B. in einem Polymer einzukapseln, um ihn vor mechanischen Einflüssen und Umwelteinflüssen zu schützen. Eine Möglichkeit zur Einkapselung ist das koaxiale Elektrospinnen. Vorteile sind unter anderem die Realisierung einer großen Oberfläche und einer sehr geringen Wanddicke der schützenden Schale, die die Diffusion von Gasen durch die Wand hindurch ermöglicht. Um die Funktionsfähigkeit eines solchen Sensors zu testen, wurde ein CO2-sensitiver Flüssigkristall verwendet. Dieser wurde in eine Schale aus Polyvinylpyrrolidon (PVP) versponnen und die Reaktion mit CO2 spektroskopisch analysiert.
Optical gas sensors based on chiral-nematic liquid crystals (N* LCs) forming one-dimensional photonic crystals do not require electrical energy and have a considerable potential to supplement established types of sensors. A chiral-nematic phase with tunable selective reflection is induced in a nematic host LC by adding reactive chiral dopants. The selective chemical reaction between dopant and analyte is capable to vary the pitch length (the lattice constant) of the soft, self-assembled, one-dimensional photonic crystal. The progress of the ongoing chemical reaction can be observed even by naked eye because the color of the samples varies. In this work, we encapsulate the responsive N* LC in microscale polyvinylpyrrolidone (PVP) fibers via coaxial electrospinning. The sensor is, thus, given a solid form and has an improved stability against nonavoidable environmental influences. The reaction behavior of encapsulated and nonencapsulated N* LC toward a gaseous analyte is compared, systematically. Making use of the encapsulation is an important step to improve the applicability.
The analysis of Δ9-tetrahydrocannabinol (THC) and its metabolites 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC), and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) from blood serum is a routine task in forensic toxicology laboratories. For examination of consumption habits, the concentration of the phase I metabolite THC-COOH is used. Recommendations for interpretation of analysis values in medical-psychological assessments (regranting of driver’s licenses, Germany) include threshold values for the free, unconjugated THC-COOH. Using a fully automated two-step liquid-liquid extraction, THC, 11-OH-THC, and free, unconjugated THC-COOH were extracted from blood serum, silylated with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), and analyzed by GC/MS. The automation was carried out by an x-y-z sample robot equipped with modules for shaking, centrifugation, and solvent evaporation. This method was based on a previously developed manual sample preparation method. Validation guidelines of the Society of Toxicological and Forensic Chemistry (GTFCh) were fulfilled for both methods, at which the focus of this article is the automated one. Limits of detection and quantification for THC were 0.3 and 0.6 μg/L, for 11-OH-THC were 0.1 and 0.8 μg/L, and for THC-COOH were 0.3 and 1.1 μg/L, when extracting only 0.5 mL of blood serum. Therefore, the required limit of quantification for THC of 1 μg/L in driving under the influence of cannabis cases in Germany (and other countries) can be reached and the method can be employed in that context. Real and external control samples were analyzed, and a round robin test was passed successfully. To date, the method is employed in the Institute of Legal Medicine in Giessen, Germany, in daily routine. Automation helps in avoiding errors during sample preparation and reduces the workload of the laboratory personnel. Due to its flexibility, the analysis system can be employed for other liquid-liquid extractions as well. To the best of our knowledge, this is the first publication on a comprehensively automated classical liquid-liquid extraction workflow in the field of forensic toxicological analysis.
DT-13 attenuates inflammation by inhibiting NLRP3-inflammasome related genes in RAW264.7 macrophages
(2024)
Plant derived saponins or other glycosides are widely used for their anti-inflammatory, antioxidant, and anti-viral properties in therapeutic medicine. In this study, we focus on understanding the function of the less known steroidal saponin from the roots of Liriope muscari L. H. Bailey – saponin C (also known as DT-13) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in comparison to the well-known saponin ginsenoside Rk1 and anti-inflammatory drug dexamethasone. We proved that DT-13 reduces LPS-induced inflammation by inhibiting nitric oxide (NO) production, interleukin-6 (IL-6) release, cycloxygenase-2 (COX-2), tumour necrosis factor-alpha (TNF-α) gene expression, and nuclear factor kappa-B (NFκB) translocation into the nucleus. It also inhibits the inflammasome component NOD-like receptor family pyrin domain containing protein 3 (NLRP3) regulating the inflammasome activation. This was supported by the significant inhibition of caspase-1 and interleukin-1 beta (IL-1β) expression and release. This study demonstrates the anti-inflammatory effect of saponins on LPS-stimulated macrophages. For the first time, an in vitro study shows the attenuating effect of DT-13 on NLRP3-inflammasome activation. In comparison to the existing anti-inflammatory drug, dexamethasone, and triterpenoid saponin Rk1, DT-13 more efficiently inhibits inflammation in the applied cell culture model. Therefore, DT-13 may serve as a lead compound for the development of new more effective anti-inflammatory drugs with minimised side effects.
Matrix metalloproteinases (MMPs) are matrix-degrading enzymes that are over-expressed in joints of rheumatoid arthritis (RA) patients. However, the contribution of specific MMPs for the development of arthritic joints is unknown. This study is aimed at studying the role of matrix metalloproteinase-9 (MMP-9) in mice, using the K/BxN serum-transfer model of RA. Arthritis was induced in Balb/c mice by injecting K/BxN serum. Development of arthritis was followed in these mice by measuring ankle thickness and clinical index score. MMP-9 expression in the joints of mice killed at various time points during the disease progression was determined by gelatin zymography using ankle lysates. We found that MMP-9 expression increased with the severity of arthritis. Importantly MMP-9 deficient mice injected with K/BxN serum showed a milder form of arthritis in comparison to the control C57BL/6 mice injected with K/BxN serum. We therefore conclude that MMP-9 promotes arthritis in mice.
Background: Migration of mature and immature leukocytes in response to chemokines is not only essential during inflammation and host defense, but also during development of the hematopoietic system. Many molecules implicated in migratory polarity show uniform cellular distribution under non-activated conditions, but acquire a polarized localization upon exposure to migratory cues.
Methodology/Principal Findings: Here, we present evidence that raft-associated endocytic proteins (flotillins) are preassembled in lymphoid, myeloid and primitive hematopoietic cells and accumulate in the uropod during migration. Furthermore, flotillins display a polarized distribution during immunological synapse formation. Employing the membrane lipid-order sensitive probe Laurdan, we show that flotillin accumulation in the immunological synapse is concomittant with membrane ordering in these regions.
Conclusions: Together with the observation that flotillin polarization does not occur in other polarized cell types such as polarized epithelial cells, our results suggest a specific role for flotillins in hematopoietic cell polarization. Based on our results, we propose that in hematopoietic cells, flotillins provide intrinsic cues that govern segregation of certain microdomain-associated molecules during immune cell polarization.
Process-dependent thermo-mechanical viscoelastic properties and the corresponding morphology of HDPE extrusion blow molded (EBM) parts were investigated. Evaluation of bulk data showed that flow direction, draw ratio, and mold temperature influence the viscoelastic behavior significantly in certain temperature ranges. Flow induced orientations due to higher draw ratio and higher mold temperature lead to higher crystallinities. To determine the local viscoelastic properties, a new microindentation system was developed by merging indentation with dynamic mechanical analysis. The local process-structure-property relationship of EBM parts showed that the cross-sectional temperature distribution is clearly reflected by local crystallinities and local complex moduli. Additionally, a model to calculate three-dimensional anisotropic coefficients of thermal expansion as a function of the process dependent crystallinity was developed based on an elementary volume unit cell with stacked layers of amorphous phase and crystalline lamellae. Good agreement of the predicted thermal expansion coefficients with measured ones was found up to a temperature of 70 °C.
This study presents a microindentation system which allows spatially resolved local as well as bulk viscoelastic material information to be obtained within one instrument. The microindentation method was merged with dynamic mechanical analysis (DMA) for a tungsten cone indenter. Three tungsten cone indenters were investigated: tungsten electrode, tungsten electrode + 2% lanthanum, and tungsten electrode + rare earth elements. Only the tungsten electrode + 2% lanthanum indenter showed the sinusoidal response, and its geometry remained unaffected by the repeated indentations. Complex moduli obtained from dynamic microindentation for high-density polyethylene, polybutylene terephthalate, polycarbonate, and thermoplastic polyurethane are in agreement with the literature. Additionally, by implementing a specially developed x-y-stage, this study showed that dynamic microindentation with a tungsten cone indenter was an adequate method to determine spatially resolved local viscoelastic surface properties.
Influence of design of extrusion blow molding (EBM) in terms of extrusion direction set-up and draw ratio as well as process conditions (mold temperature) on storage modulus of high density polyethylene EBM containers was analyzed with dynamic mechanical analysis. All three parameters - mold temperature, flow direction and draw ratio - are statistically significant and lead to relative and absolute evaluation of storage modulus. Furthermore, flow induced changes in crystallinity was analyzed by differential scanning calorimetry. Obtained data on deformation properties can be employed for more sophisticated finite element simulations with the aim to reach more sustainable extrusion blow molding production.
Ultra-fast photopolymerization of experimental composites: DEA and FT-NIRS measurement comparison
(2015)
Introduction of Matrix-Filler Adhesion to Modelling of Elastic Moduli of Particulate Composites
(2022)
Cube in cube elementary volume (EV) concept serves to predict a filler-content dependent Young´s moduli of particle filled composites using moduli of a matrix EM and a filler EF. Paul and Ishai-Cohen derived formulas for composites moduli considering different load transfer boundaries in the EV assuming a complete filler-matrix adhesion. In this paper it is confirmed that their models represent the upper and lower bounds, respectively, with the respect to the experimental data. However, in vast majority of composites a filler-matrix adhesion is not complete. Therefore, an adhesion factor kadh gaining values between 0 and 1 was introduced into Paul´s model to consider the reduced adhesion as the reduction of the filler-matrix contact area for glass beads filled in polar and unpolar thermoplastic matrices as well as elastomer. The evaluation of these composite systems provides reasonable adhesion coefficients of PA66 > PBT > PP > PE-LD >> BR. It was also found that stiffening only occurs if kadh exceeds the minimum value adhesion of root square of E(M) divided by E(F). The determined kadh correspond to scanning electron microscopy observations of the composites fracture surfaces. Additionally, finite element analysis of the cubic and hexagonal arrangements of the EV show that the stress distributions are different, but they affect the calculated moduli only for the filler volume contents exceeding 20 %. The introduction of the filler-matrix adhesion provides more reliable predictions of Young´s moduli of particulate composites.
Modeling of Creep Behavior of Particulate Composites with Focus on Interfacial Adhesion Effect
(2022)
Evaluation of creep compliance of particulate composites using empirical models always provides parameters depending on initial stress and material composition. The effort spent to connect model parameters with physical properties has not resulted in success yet. Further, during the creep, delamination between matrix and filler may occur depending on time and initial stress, reducing an interface adhesion and load transfer to filler particles. In this paper, the creep compliance curves of glass beads reinforced poly(butylene terephthalate) composites were fitted with Burgers and Findley models providing different sets of time-dependent model parameters for each initial stress. Despite the finding that the Findley model performs well in a primary creep, the Burgers model is more suitable if secondary creep comes into play; they allow only for a qualitative prediction of creep behavior because the interface adhesion and its time dependency is an implicit, hidden parameter. As Young’s modulus is a parameter of these models (and the majority of other creep models), it was selected to be introduced as a filler content-dependent parameter with the help of the cube in cube elementary volume approach of Paul. The analysis led to the time-dependent creep compliance that depends only on the time-dependent creep of the matrix and the normalized particle distance (or the filler volume content), and it allowed accounting for the adhesion effect. Comparison with the experimental data confirmed that the elementary volume-based creep compliance function can be used to predict the realistic creep behavior of particulate composites.
Apple replant disease (ARD) is a soil-borne disease, which is of particular importance for fruit tree nurseries and fruit growers. The disease manifests by a poor vegetative development, stunted growth, and reduced yield in terms of quantity and quality, if apple plants (usually rootstocks) are replanted several times at the same site. Genotype-specific differences in the reaction of apple plants to ARD are documented, but less is known about the genetic mechanisms behind this symptomatology. Recent transcriptome analyses resulted in a number of candidate genes possibly involved in the plant response. In the present study, the expression of 108 selected candidate genes was investigated in root and leaf tissue of four different apple genotypes grown in untreated ARD soil and ARD soil disinfected by γ-irradiation originating from two different sites in Germany. Thirty-nine out of the 108 candidate genes were differentially expressed in roots by taking a p-value of < 0.05 and a fold change of > 1.5 as cutoff. Sixteen genes were more than 4.5-fold upregulated in roots of plants grown in ARD soil. The four genes MNL2 (putative mannosidase); ALF5 (multi antimicrobial extrusion protein); UGT73B4 (uridine diphosphate (UDP)-glycosyltransferase 73B4), and ECHI (chitin-binding) were significantly upregulated in roots. These genes seem to be related to the host plant response to ARD, although they have never been described in this context before. Six of the highly upregulated genes belong to the phytoalexin biosynthesis pathway. Their genotype-specific gene expression pattern was consistent with the phytoalexin content measured in roots. The biphenyl synthase (BIS) genes were found to be useful as early biomarkers for ARD, because their expression pattern correlated well with the phenotypic reaction of the Malus genotypes investigated.
Analytische Chemie I
(2019)
Analytical Chemistry I
(2023)
This workbook takes you through the successful work Harris, Textbook of Quantitative Analysis and is designed primarily for self-study. In five parts, the lecture content of analytical chemistry is summarized and explained using selected examples. Basic concepts of analytical chemistry are presented as well as the principle and various techniques of dimensional analysis and chromatography. UV/VIS, infrared and Raman spectroscopy are used to explain the investigation of molecularly present compounds, and selected techniques of atomic spectroscopy conclude the introduction to the fundamentals of analysis. The textbook's essential sections and illustrations are repeatedly referred to, which facilitates independent learning of the fundamentals of analytical chemistry.
Analytische Chemie II
(2020)
Dieses Arbeitsbuch führt durch das erfolgreiche Lehrbuch Skoog/Holler/Crouch, Instrumentelle Analytik und ist vor allem für das Selbststudium konzipiert. In fünf Teilen werden die Vorlesungsinhalte der fortgeschritteneren Analytischen Chemie zusammengefasst und anhand ausgewählter Beispiele erläutert: Mit der Untersuchung von Molekülen befassen sich Massenspektrometrie und Kernresonanzspektroskopie, zudem werden zahlreiche elektroanalytische Methoden wie Potentiometrie, Coulometrie, Amperometrie und Voltammetrie behandelt. In einem Überblick über speziellere Verfahren der Analytik geht es unter anderem ebenso um den Einsatz radioaktiver Substanzen und die Nutzung verschiedener Fluoreszenzverfahren wie um Methoden der Informationsgewinnung in der zunehmend wichtigen elektrochemischen und optischen Sensortechnik sowie deren Automatisierbarkeit. Den Abschluss bildet eine Zusammenfassung verschiedener Prinzipien und Anwendungsmethoden der Statistik, die im Rahmen der Analytik schlichtweg unverzichtbar sind. Um das selbstständige Lernen zu erleichtern, wird dabei in allen Teilen des Buches immer wieder auf essenzielle Abschnitte und Abbildungen des Lehrbuches verwiesen.
Die Analytik ist zwar ein grundlegendes Thema innerhalb der Chemie, hat es aber ganz schön in sich. Aber keine Sorge, Ulf Ritgen erklärt Ihnen in diesem Buch was Sie zur Analytischen Chemie Wissen sollten. Wie erhält man eigentlich analytische Infos? Welche Standards gibt es? Und wie sollte man überhaupt mit den Stoffgemischen umgehen? All diese Grundlagen werden ausführlich erläutert. Aber auch die Anwendungsgebiete und Methoden kommen keinesfalls zu kurz. Egal ob Gravimetrie, Titration, Fällung oder Konduktometrie, endlich wird alles verständlich erklärt. Jetzt können Prüfung und Praktikum kommen! (Verlagsangaben)
A main factor hampering life in space is represented by high atomic number nuclei and energy (HZE) ions that constitute about 1% of the galactic cosmic rays. In the frame of the “STARLIFE” project, we accessed the Heavy Ion Medical Accelerator (HIMAC) facility of the National Institute of Radiological Sciences (NIRS) in Chiba, Japan. By means of this facility, the extremophilic species Haloterrigena hispanica and Parageobacillus thermantarcticus were irradiated with high LET ions (i.e., Fe, Ar, and He ions) at doses corresponding to long permanence in the space environment. The survivability of HZE-treated cells depended upon either the storage time and the hydration state during irradiation; indeed, dry samples were shown to be more resistant than hydrated ones. With particular regard to spores of the species P. thermantarcticus, they were the most resistant to irradiation in a water medium: an analysis of the changes in their biochemical fingerprinting during irradiation showed that, below the survivability threshold, the spores undergo to a germination-like process, while for higher doses, inactivation takes place as a consequence of the concomitant release of the core’s content and a loss of integrity of the main cellular components. Overall, the results reported here suggest that the selected extremophilic microorganisms could serve as biological model for space simulation and/or real space condition exposure, since they showed good resistance to ionizing radiation exposure and were able to resume cellular growth after long-term storage.
We examined the cytotoxic effects of chaetocin on clear cell renal cell carcinoma (ccRCC) cells and the possibility to combine the effects of chaetocin with the effects of cytokine-induced killer cells (CIK) assayed by MTT assay and FACS analysis. Chaetocin is a thiodioxopiperazine produced by fungi belonging to the chaetomiaceae family. In 2007, it was first reported that chaetocin shows potent and selective ex vivo anti-cancer activity by inducing reactive oxygen species. CIK cells are generated from CD3+/CD56- T lymphocytes with double negative CD4-/CD8- phenotype that are isolated from human blood. The addition of distinct interleukins and antibodies results in the generation of CIK cells that are able to specifically target and destroy renal carcinoma cells. The results of this research state that the anti-ccRCC activity of chaetocin is weak and does not show a high grade of selectivity on clear cell renal cell carcinoma cells. Although the CIK cells show a high grade of selective anti-ccRCC activity, this effect could not be improved by the addition of chaetocin. So chaetocin seems to be no suitable agent for specific targeting ccRCC cells or for the combination therapy with CIK cells in renal cancer.
Lignin ist ein aromatisches Biopolymer, das in den Zellwänden von Pflanzen vorkommt. Es ist hauptsächlich aus drei sogenannten Monolignolen (p-Hydroxyphenyl (H), Guajakol (G) und Syringol (S)) aufgebaut, die über verschiedene Bindungen miteinander verknüpft sein können, und enthält eine Vielzahl an funktionellen Gruppen. Interessant für die Verwendung von Lignin sind dabei insbesondere die vielen phenolischen Hydroxygruppen, die als Ausgangsstoff bei der Synthese neuer Produkte dienen können, daneben aber auch für seine antioxidativen Eigenschaften verantwortlich sind. Da Struktur und Eigenschaften von vielen Faktoren wie Biomasse und Aufschlussprozess abhängen, ist eine detaillierte Charakterisierung der Lignine nötig, um Struktur-Eigenschafts-Beziehungen aufzuklären und so einen Schritt näher an eine mögliche stoffliche Nutzung zu kommen. Mit dieser Arbeit soll der Einfluss der Biomasse inklusive der verwendeten Partikelgröße sowie des Organosolv-Aufschlussprozesses auf die Monomerzusammensetzung, das Molekulargewicht und die Antioxidanz der isolierten Lignine untersucht werden.
Als Rohstoffe zur Ligningewinnung dienen die drei mehrjährigen lignocellulosereichen Low-Input-Pflanzen Miscanthus x giganteus, Silphium perfoliatum und Paulownia tomentosa, die momentan hauptsächlich zur Energiegewinnung genutzt werden. Im Rahmen der Bioökonomiestrategie der Europäischen Union soll der Schwerpunkt zukünftiger Bioraffinerien jedoch auf eine ganzheitliche Nutzung von Biomassen gelegt und so auch die stoffliche Nutzung fokussiert werden. Zusätzlich zu diesen drei Pflanzen werden auch Organosolv-Lignine aus den in der Literatur bereits gut beschriebenen Biomassen Weizenstroh und Buchenholz isoliert, und zwei Nadelholz-Kraft-Lignine als Vergleich herangezogen. Die Ergebnisse zeigen, dass die Art der Biomasse hauptsächlich die Monomerzusammensetzung beeinflusst: Gräser bestehen aus allen drei Monolignolen, Laubhölzer mehrheitlich aus S- und G-Einheiten, während Nadelhölzer nur aus G-Einheiten aufgebaut sind. Die Holzlignine besitzen zudem höhere Molekulargewichte sowie bessere antioxidative Eigenschaften als die Gras- und Krautlignine. Mit der feineren Vermahlung der Biomasse kann die Monomerzusammensetzung beeinflusst werden: der Einsatz kleinerer Partikelgrößen führt zu Ligninen mit einem höheren Gehalt an H-Einheiten, sowohl für Miscanthus als auch für Paulownia. Außerdem kann bei Paulownia die Ausbeute gesteigert und eine Zunahme des Molekulargewichtes beobachtet werden, wenn die kleinste Siebfraktion für den Organosolv-Aufschluss verwendet wird. Einen größeren Einfluss als der Mahlgrad der Biomasse haben die Autohydrolyse sowie der Organosolv-Aufschlussprozess selbst. Die Monomerzusammensetzung ändert sich aufgrund derselben Biomasse zwar kaum, die Bindungstypen zwischen den Monolignolen dagegen schon. Mit höherer Prozessstärke (Zeit, Temperatur, Ethanol-Konzentration) werden Etherbindungen gespalten, was den Anteil an phenolischen Hydroxygruppen und somit die Antioxidanz erhöht. Neben dieser Depolymerisation werden partiell auch Rekondensationsreaktionen beobachtet.
Die erzielten Ergebnisse liefern einen Beitrag zum Verständnis des Zusammenhangs zwischen Ligninquelle und -gewinnung mit der daraus resultierenden Ligninstruktur und Antioxidanz und bieten damit eine Grundlage für den Wandel von der energetischen hin zu einer nachhaltigen stofflichen Nutzung dieses nachwachsenden Biopolymers. Gerade über die Wahl der Aufschlussparameter können Struktur und Antioxidanz gezielt beeinflusst werden, was in zukünftigen Studien weiter fokussiert werden sollte.
This research studies in detail four different assays, namely DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), FRAP (ferric ion reducing antioxidant potential) and FC (Folin-Ciocalteu), to determine the antioxidant capacity of standard substances as well as 50 organosolv lignins, and two kraft lignins. The coefficient of variation was determined for each method and was lowest for ABTS and highest for DPPH. The best correlation was found for FRAP and FC, which both rely on a single electron transfer mechanism. A good correlation between ABTS, FRAP and FC, respectively, could be observed, even though ABTS relies on a more complex reaction mechanism. The DPPH assay merely correlates with the others, implying that it reflects different antioxidative attributes due to a different reaction mechanism. Lignins obtained from paulownia and silphium have been investigated for the first time regarding their antioxidant capacity. Paulownia lignin is in the same range as beech wood lignin, while silphium lignin resembles wheat straw lignin. Miscanthus lignin is an exception from the grass lignins and possesses a significantly higher antioxidant capacity. All lignins possess a good antioxidant capacity and thus are promising candidates for various applications, e. g. as additives in food packaging or for biomedical purposes.
Streptococcus agalactiae is the leading cause of bacterial sepsis and meningitis in neonates and is also the causative agent of several serious infections in immunocompromised adults. S. agalactiae encounters multiple niches during an infection, suggesting that regulatory mechanisms control the expression of specific virulence factors in this bacterium. The present study describes the functional characterization of a gene from S. agalactiae, designated rga, which encodes a protein with significant similarity to members of the RofA-like protein (RALP) family of transcriptional regulators. After deletion of the rga gene in the genome of S. agalactiae, the mutant strain exhibited significantly reduced expression of the genes srr-1 and pilA, which encode a serine-rich repeat surface glycoprotein and a pilus protein, respectively, and moderately increased expression of the fbsA gene, which encodes a fibrinogen-binding protein. Electrophoretic mobility shift assays demonstrated specific DNA binding of purified Rga to the promoter regions of pilA and fbsA, suggesting that Rga directly controls pilA and fbsA. Adherence assays revealed significantly reduced binding of the Δrga mutant to epithelial HEp-2 cells and to immobilized human keratin 4, respectively. In contrast, the adherence of the Δrga mutant to A549 cells and its binding to human fibrinogen was significantly increased. Immunoblot and immunoelectron microscopy revealed that the quantity of pilus structures was significantly reduced in the Δrga mutant compared with the parental strain. The wild-type phenotype could be restored by plasmid-mediated expression of rga, demonstrating that the mutant phenotypes resulted from a loss of Rga function.
Pozzolanic properties of Pennisetum purpureum grass ash were tested on Portland cement. Results show that the ash can be blended with cements without compromising binding strength of the cement. It was found that Portland cement could be blended with Pennisetum purpureum up to a ratio of 3:2 compromising compressive strength of mortar.Mortar with lower cement replacement took longer to set as evidenced by lower compressive strength within the 28-day aging time. Mortar with higher cement replacement had lower water absorption capacity, an indication that the test pozzolan was of smaller particulate size. XRF analysis and the FTIR spectrum showed that the ash has a higher content of silica. The XRD pattern of the ash showed that the ash was predominantly amorphous. SEM images showed that the ash produced at 600 o C had residual carbon material.
3-Hydroxy-3-methylglutaryl-coenzyme A lyase (HMGCL, HMGCL) deficiency is a rare inborn error of ketogenesis. Even if the ketogenic enzyme is fully disrupted, an elevated signal for the ketone body acetoacetic acid is a frequent observation in the analysis of urinary organic acids, at least if derivatization is performed by methylation. We provide an explanation for this phenomenon and trace it back to degradation of the derivatized 3-hydroxy-3-methylglutaric acid and high temperature of the injector of the gas chromatograph.
Major progress occurred in understanding inborn errors of ketone body transport and metabolism between the International Congresses on Inborn Errors of Metabolism in Barcelona (2013) and Rio de Janeiro (2017). These conditions impair either ketogenesis (presenting as episodes of hypoketotic hypoglycemia) or ketolysis (presenting as ketoacidotic episodes); for both groups, immediate intravenous glucose administration is the most critical and (mHGGCS, HMGCS2) effective treatment measure.
In memoriam Willy Lehnert
(2023)
Polymer fibers with liquid crystals (LCs) in the core have potential as autonomous sensors of airborne volatile organic compounds (VOCs), with a high surface-to-volume ratio enabling fast and sensitive response and an attractive non-woven textile form factor. We demonstrate their ability to continuously and quantitatively measure the concentration of toluene, cyclohexane, and isopropanol as representative VOCs, via the impact of each VOC on the LC birefringence. The response is fully reversible and repeatable over several cycles, the response time can be as low as seconds, and high sensitivity is achieved when the operating temperature is near the LC-isotropic transition temperature. We propose that a broad operating temperature range can be realized by combining fibers with different LC mixtures, yielding autonomous VOC sensors suitable for integration in apparel or in furniture that can compete with existing consumer-grade electronic VOC sensors in terms of sensitivity and response speed.
Suprabasal BCL-2 Expression Does Not Sensitize to Chemically-induced Skin Cancer in Transgenic Mice
(2008)
Nitrile-type inhibitors are known to interact with cysteine proteases in a covalent-reversible manner. The chemotype of 3-cyano-3-aza-β-amino acid derivatives was designed in which the N-cyano group is centrally arranged in the molecule to allow for interactions with the nonprimed and primed binding regions of the target enzymes. These compounds were evaluated as inhibitors of the human cysteine cathepsins K, S, B, and L. They exhibited slow-binding behavior and were found to be exceptionally potent, in particular toward cathepsin K, with second-order rate constants up to 52 900 × 103 M–1 s–1.
The human enzymes GLYAT (glycine N-acyltransferase), GLYATL1 (glutamine N-phenylacetyltransferase) and GLYATL2 (glycine N-acyltransferase-like protein 2) are not only important in the detoxification of xenobiotics via the human liver, but are also involved in the elimination of acyl residues that accumulate in the form of their coenzyme A (coA) esters in some rare inborn errors of metabolism. This concerns, for example, disorders in the degradation of branched-chain amino acids, such as isovaleric acidemia or propionic acidemia. In addition, they also assist in the elimination of ammonium, which is produced during the transamination of amino acids and accumulates in urea cycle defects. Sequence variants of the enzymes have also been investigated, which may provide evidence of impaired enzyme activities, from which therapy adjustments can potentially be derived. A modified Escherichia coli strain was chosen for the overexpression and partial biochemical characterization of the enzymes, which may allow solubility and proper folding. Since post-translational protein modifications are very limited in bacteria, we also attempted to overexpress the enzymes in HEK293 cells (human-derived). In addition to characterization via immunoblots and activity assays, intracellular localization of the enzymes was also performed using GFP coupling and confocal laser scanning microscopy in transfected HEK293 cells. The GLYATL2 enzyme may have tasks beyond detoxification and metabolic defects and the preliminary molecular biology work has been performed as part of this project - the enzyme activity determinations were outsourced to a co-supervised bachelor thesis. The enzyme activity determinations with purified recombinant human enzyme from Escherichia coli provided a threefold higher activity of the sequence variant p.(Asn156Ser) for GLYAT, which should be considered as the probably authentic wild type of the enzyme. In addition, a reduced activity of the GLYAT variant p.(Gln61Leu), which is very common in South Africa, was shown, which could be of particular importance in the treatment of isovaleric acidemia, which is also common in South Africa. Intracellularly, GLYAT and GLYATL1 could be localized mitochondrially. As the analyses have shown, sequence variations of GLYAT and GLYATL1 influence their enzyme activity. As an example, the GLYAT variant p.(Gln61Leu) is frequently found in South Africa. In the case of reduced GLYAT activity, patients could be increasingly treated with L-carnitine in the sense of an individualized therapy, since the conjugation of the toxic isovaleryl-coA with glycine is restricted by the GLYAT sequence variation. Activity-reducing variants identified in this project are of particular interest, as they may influence the treatment of certain metabolic defects.
Biomass in general, wood and grasses in particular represent attractive renewable sources for the fabrication of so-called building block chemicals (1). Thus, environmentally benign antimicrobial nanoparticles based on a silver-infused lignin core were recently reported underlying the high potential for valorization of lignin (2). The contribution presents specific correlations regarding the structural differences of lignins depending on both: source (wood vs. grass) and isolation procedure (Kraft vs. Organosolv). Special focus will be drawn on detailed structure deviations caused by Miscanthus genotypes (M. gigantheus, M. robustus, M. sisnensis).