Changing mechanical properties of photopolymerized, dityrosine-crosslinked protein-based hydrogels

Hydrogels based on renewable resources are a promising class of materials for future applications in pharmaceutics, drug delivery and personalized medicine. Thus, optional adjustments of mechanical properties such as swelling behavior, elasticity and network strength are desired. In this context, hydrogels based on the biological raw materials bovine serum albumin and casein were prepared by dityrosine-crosslinking of their tyrosine residues through visible light-induced photopolymerization. Changing the tyrosine accessibility by urea addition before photopolymerization increased the storage modulus of the hydrogels by 650% while simultaneously being more elastic. Furthermore, contributions of the buffer system composition, variation of protein concentration and storage medium towards mechanical properties of the hydrogel such as storage moduli, elasticity, fracture strain, compressive strength and relative weight swelling ratio are discussed. It could be shown, that changes in precursor solution and storage medium characteristics are crucial parameters towards tuning the mechanical properties of protein-based hydrogels

Diclofenac and other non-steroidal anti-inflammatory drugs (NSAIDs) are competitive antagonists of the human P2X3 receptor

Introduction: The P2X3 receptor (P2X3R), an ATP-gated non-selective cation channel of the P2X receptor family, is expressed in sensory neurons and involved in nociception. P2X3R inhibition was shown to reduce chronic and neuropathic pain. In a previous screening of 2000 approved drugs, natural products, and bioactive substances, various non-steroidal anti-inflammatory drugs (NSAIDs) were found to inhibit P2X3R-mediated currents.Methods: To investigate whether the inhibition of P2X receptors contributes to the analgesic effect of NSAIDs, we characterized the potency and selectivity of various NSAIDs at P2X3R and other P2XR subtypes using two-electrode voltage clamp electrophysiology.Results: We identified diclofenac as a hP2X3R and hP2X2/3R antagonist with micromolar potency (with IC50 values of 138.2 and 76.7 µM, respectively). A weaker inhibition of hP2X1R, hP2X4R, and hP2X7R by diclofenac was determined. Flufenamic acid (FFA) inhibited hP2X3R, rP2X3R, and hP2X7R (IC50 values of 221 µM, 264.1 µM, and ∼900 µM, respectively), calling into question its use as a non-selective ion channel blocker, when P2XR-mediated currents are under study. Inhibition of hP2X3R or hP2X2/3R by diclofenac could be overcome by prolonged ATP application or increasing concentrations of the agonist α,β-meATP, respectively, indicating competition of diclofenac and the agonists. Molecular dynamics simulation showed that diclofenac largely overlaps with ATP bound to the open state of the hP2X3R. Our results suggest a competitive antagonism through which diclofenac, by interacting with residues of the ATP-binding site, left flipper, and dorsal fin domains, inhibits the gating of P2X3R by conformational fixation of the left flipper and dorsal fin domains. In summary, we demonstrate the inhibition of the human P2X3 receptor by various NSAIDs. Diclofenac proved to be the most effective antagonist with a strong inhibition of hP2X3R and hP2X2/3R and a weaker inhibition of hP2X1R, hP2X4R, and hP2X7R.Discussion: Considering their involvement in nociception, inhibition of hP2X3R and hP2X2/3R by micromolar concentrations of diclofenac, which are rarely reached in the therapeutic range, may play a minor role in analgesia compared to the high-potency cyclooxygenase inhibition but may explain the known side effect of taste disturbances caused by diclofenac

Clinical and virological characteristics of hospitalised COVID-19 patients in a German tertiary care centre during the first wave of the SARS-CoV-2 pandemic: a prospective observational study

Purpose: Adequate patient allocation is pivotal for optimal resource management in strained healthcare systems, and requires detailed knowledge of clinical and virological disease trajectories. The purpose of this work was to identify risk factors associated with need for invasive mechanical ventilation (IMV), to analyse viral kinetics in patients with and without IMV and to provide a comprehensive description of clinical course. Methods: A cohort of 168 hospitalised adult COVID-19 patients enrolled in a prospective observational study at a large European tertiary care centre was analysed. Results: Forty-four per cent (71/161) of patients required invasive mechanical ventilation (IMV). Shorter duration of symptoms before admission (aOR 1.22 per day less, 95% CI 1.10-1.37, p < 0.01) and history of hypertension (aOR 5.55, 95% CI 2.00-16.82, p < 0.01) were associated with need for IMV. Patients on IMV had higher maximal concentrations, slower decline rates, and longer shedding of SARS-CoV-2 than non-IMV patients (33 days, IQR 26-46.75, vs 18 days, IQR 16-46.75, respectively, p < 0.01). Median duration of hospitalisation was 9 days (IQR 6-15.5) for non-IMV and 49.5 days (IQR 36.8-82.5) for IMV patients. Conclusions: Our results indicate a short duration of symptoms before admission as a risk factor for severe disease that merits further investigation and different viral load kinetics in severely affected patients. Median duration of hospitalisation of IMV patients was longer than described for acute respiratory distress syndrome unrelated to COVID-19

Novel loci for childhood body mass index and shared heritability with adult cardiometabolic traits

The genetic background of childhood body mass index (BMI), and the extent to which the well-known associations of childhood BMI with adult diseases are explained by shared genetic factors, are largely unknown. We performed a genome-wide association study meta-analysis of BMI in 61,111 children aged between 2 and 10 years. Twenty-five independent loci reached genome-wide significance in the combined discovery and replication analyses. Two of these, located nearNEDD4LandSLC45A3, have not previously been reported in relation to either childhood or adult BMI. Positive genetic correlations of childhood BMI with birth weight and adult BMI, waist-to-hip ratio, diastolic blood pressure and type 2 diabetes were detected (R(g)ranging from 0.11 to 0.76, P-values Author summary Although twin studies have shown that body mass index (BMI) is highly heritable, many common genetic variants involved in the development of BMI have not yet been identified, especially in children. We studied associations of more than 40 million genetic variants with childhood BMI in 61,111 children aged between 2 and 10 years. We identified 25 genetic variants that were associated with childhood BMI. Two of these have not been implicated for BMI previously, located close to the genesNEDD4LandSLC45A3. We also show that the genetic background of childhood BMI overlaps with that of birth weight, adult BMI, waist-to-hip-ratio, diastolic blood pressure, type 2 diabetes, and age at menarche. Our results suggest that the biological processes underlying childhood BMI largely overlap with those underlying adult BMI. However, the overlap is not complete. Additionally, the genetic backgrounds of childhood BMI and other cardio-metabolic phenotypes are overlapping. This may mean that the associations of childhood BMI and later cardio-metabolic outcomes are partially explained by shared genetics, but it could also be explained by the strong association of childhood BMI with adult BMI.Peer reviewe

Nutzung regionaler abfall- und abwasserstämmiger Bioressourcen am Beispiel des Hamburger Demonstrationsvorhabens „Jenfelder Au“ : Inventur, Lagerung, Aufbereitung, Vergärung und Gärrestverwertung

Im vorliegenden Forschungsbericht werden Arbeiten und Ergebnisse vorgestellt, die im Rahmen des vom BMBF geförderten Verbundprojektes KREIS (Kopplung von regenerativer Energiegewinnung mit innovativer Stadtentwässerung) erstellt wurden (Förderkennzeichen 033L047C). KREIS bereitete die Umsetzung des von Hamburg Wasser entwickelten Hamburg Water Cycle® (HWC) in einem Neubaugebiet in Hamburg vor. Der HWC sieht die getrennte Erfassung von Regen-, Grau- und Schwarzwasser und eine dezentrale Behandlung der drei Abwasserströme vor. Die Arbeiten der TUHH umfassten dabei den Bereich der Schwarzwasserverwertung in Verbindung mit weiteren regionalen Reststoffen. Konzentriertes Schwarzwasser aus Vakuumtoiletten wurde mit Co-Substraten anaerob zu Biogas umgesetzt, welches Strom sowie Wärme für den Stadtteil liefert. Verwertungsmöglichkeiten für den verbleibenden Gärrest wurden betrachtet. Zunächst wurde eine Inventur mögliche regionaler Co-Substrate durchgeführt. Rasenschnitt und Fettwässer wurden ausgewählt und hinsichtlich Trockenrückstand, Organik, Gesamt-Stickstoff (N), -Kohlenstoff (C) und -Phosphor (P) charakterisiert. Untersuchungen zur Lagerung und Aufbereitung von Rasenschnitt zeigten, dass eine ganzjährige Nutzung dieser Bioressource ohne Qualitätsverluste möglich ist und sowohl als Suspension oder in Form eines Presssaftes einer Nassvergärung zugeführt werden kann. Die Biogaspotenziale des Rasenschnitts hingen stark von dessen Herkunft und Vorgeschichte ab und lagen für Presssaft bei 500-900 nL/kgoTR und für Rasenschnittsuspension bei 300-500 nL/kgoTR. In beiden Fällen wirkte sich die Zugabe positiv auf die Vergärung aus. Sowohl im Hinblick auf Prozessstabilität als Biogaserträge sind Mischungen von Schwarzwasser mit Fettwasser und einer Rasenschnittvariante gut für die Anforderungen im betrachteten Neubaugebiet geeignet. Der Gärrest der Substratmischungen beinhaltet hohe Nährstoffgehalte, die für eine Verwertung relevant sind. Untersuchungen zur Fest-Flüssig-Trennung wurden als Grundlage für eine Gärrestverwertung durchgeführt. Es wurden 3 Beschickungsszenarien für das untersuchte Gebiet betrachtet und hinsichtlich Biogasgewinnung und Nährstoffgehalten im Gärrest bewertet. Eine Substratmischung aus Schwarzwasser (40 % FM), Fettwasser (30 % FM) und Rasenschnittsilage (30 % FM) lieferte in allen Bereichen die besten Ergebnisse. Diese Mischung erreichte mit ca. 37 m³ Biogas/tFM die höchsten Erträge. Auch der Gärrest zeigte mit 0,7 g Phosphor und 2,6 g Stickstoff pro Liter Gärrest die höchsten Nährstoffgehalte. Der Gärrest hatte einen Trockenrückstand von 6,5 % FM. Er eignet sich somit gut für eine Fest-Flüssig-Trennung mittels Schneckenpresse. In einem Vergleich verschiedener Gärrestverwertungszenarien wurde der Düngewert unterschiedlicher Gärrestprodukte abgeschätzt. Die Betrachtungen zeigten, dass durch eine Kombination von energetischen und stofflichen Rückgewinnungsverfahren die Verwertungseffizienz der Reststoffe verbessert werden kann.In this research report work and results are presented, which were created as part of the BMBF funded joint project KREIS (Kopplung von regenerativer Energiegewinnung mit innovativer Stadtentwässerung) (funding code 033L047C). The aim of KREIS was to prepare for the implementation of Hamburg Water Cycle® (HWC) developed by Hamburg Wasser in a new housing estate in Hamburg. The HWC focuses on separate collection of rain, grey and blackwater and decentralized treatment of the three wastewater streams. The work carried out by the TUHH covered the research field of blackwater treatment together with regional biogenic residues. Vacuum toilets are used to collect concentrated blackwater for anaerobically treatment with waste-derived co-substrates and convert them into biogas, which is intended to supply electricity and heat for the district. Treatment possibilities for the remaining digestate were also considered. An inventory of potential co-substrates was conducted. Lawn cuttings and greasy water were selected and characterized in terms of dry residue, organics, total nitrogen (TN), total carbon (TC) and total phosphorus (TP) contents. Studies on the storage and preparation of lawn cuttings showed that this bioresource is available all year round without quality losses and can be used as a suspension or in the form of a press juice. The biogas potential depends heavily on the history of the grass with further variation between lawn cuttings and its pressed juice with a measure of 300-500 nL / kgoTR and 500-900 nL /kgoTR respectively. The addition of either lawn cuttings or press juice was observed to have a positive effect on fermentation process. In regards to process stability and biogas yields, mixtures of the blackwater with greasy water and a lawn cutting variant were well suited to the requirements in the considered living area. The digestate of the substrate mixtures contained high nutrient contents that are relevant for recovery. Investigations into the solid-liquid separation were carried out as the basis for digestate recovery. The observations showed that a combination of energetic and material recovery processes can improve the recycling efficiency of the researched residues. Three feed scenarios for the investigated area were considered and evaluated with regard to biogas production and nutrient content of the digestate. A substrate mixture of blackwater (40% FM), greasy water (30% FM) and lawn silage (30% FM) provided the best results in all areas. Firstly, this mixture achieved the highest yields with approx. 37 m³ Biogas / tFM and secondly, the digestate also contained the highest nutrient contents with 0.7 g / L phosphorus and 2.6 g / L nitrogen. Finally, the fermentation residue had a dry matter content of 6.5 % FM and is therefore well suited for a solid-liquid separation by means of screw press. In a comparison of various digestate recovery scenarios, the fertilization value of different digestate products was estimated, showing that a full recovery of resources can be achieved while simultaneously obtaining valuable nutrient products using a holistic treatment method.Bundesministerium für Bildung und Forschung (Projekt-Nr. BMBF: 033L047C

Biogas and nutrients from blackwater, lawn cuttings and grease trap residues : experiments for Hamburg’s Jenfelder Au district

Background: The project KREIS focuses on a new combination of renewable energy provision with innovative wastewater treatment, called the “Hamburg Water Cycle®” (HWC) which will be applied in Hamburg’s neighbourhood Jenfelder Au. HWC includes a separate collection of rainwater, greywater and blackwater. Vacuum toilets are used to concentrate the blackwater. Biogas will be produced from the blackwater in an anaerobic digestion process together with co-substrates. The blackwater will be transported to the anaerobic pre-treatment facility via a vacuum system. Construction of water systems started in 2013, and commercialization of houses is planned to be finished in 2018. Methods: The article focuses on research work accompanying the demonstration project. Blackwater and the co-substrates, lawn cuttings and grease trap residues from restaurants and canteens will be considered as bioresources, not as residues. To evaluate the utilization efficiency, three investigation steps were carried out: inventory to determine substrate quantities and qualities, anaerobic digestion to determine biogas production, and evaluation of digestate utilization options. Results: The daily amount of blackwater in Jenfelder Au is calculated to be about 12 m3 (dry matter (DM) 0.6 %; organic dry matter (oDM) 65 % DM; nitrogen (N) 28 % DM; phosphorus (P) 2.7 %). To increase the biogas production, co-substrates will be added. Grease trap residues (averages: DM 2 %; oDM 85 % DM; N 2.5 % DM; P 0.6 % DM) and lawn cuttings (averages: DM 30 %; oDM 80 % DM; N 2.6 % DM; P 0.3 % DM) were selected. The inventory study showed a sufficient potential of lawn cuttings within a 5-km radius. The lawn cuttings must be pre-treated for wet fermentation. Two options were investigated: press juice preparation and wet shredding of the fresh and silage lawn. Batch test was used to determine the biogas potential of the substrates with the following average results: blackwater 500 nl/kg oDM, grease trap residues 1000 nl/kg oDM, lawn cuttings 400 nl/kg oDM and lawn juice 500 nl/kg oDM. The effects of the composition of the substrate mixture and of the retention time in the reactor on biogas quantity and process stability were studied in semi-continuous operating reactors. Experiments showed that a stable process with an average biogas production of 800 nl/kg oDM is, e.g., possible with a mixture of blackwater, press juice of lawn cuttings and grease trap residues in a fresh mass ratio of 1:1:1. Furthermore, the N and P contents in digestates were determined. These nutrients are valuable for fertilization. Conclusion: It has been shown that blackwater combined with local waste streams can be used for biogas generation and that it has a potential as fertilizer. The experiments have shown that co-digestion has a positive effect on biogas yields and lawn cuttings are suitable as co-substrate. Lawn cuttings can be applied as lawn juice or lawn suspension. Ways of an integral utilization and the potential of nutrient recovery are shown in this work.Projektträger Jülich (PTJ) and the Federal Ministry of Education and Research (BMBF)

MEST mediates the impact of prenatal bisphenol A exposure on long-term body weight development

Abstract Background Exposure to endocrine-disrupting chemicals can alter normal physiology and increase susceptibility to non-communicable diseases like obesity. Especially the prenatal and early postnatal period is highly vulnerable to adverse effects by environmental exposure, promoting developmental reprogramming by epigenetic alterations. To obtain a deeper insight into the role of prenatal bisphenol A (BPA) exposure in children’s overweight development, we combine epidemiological data with experimental models and BPA-dependent DNA methylation changes. Methods BPA concentrations were measured in maternal urine samples of the LINA mother-child-study obtained during pregnancy (n = 552), and BPA-associated changes in cord blood DNA methylation were analyzed by Illumina Infinium HumanMethylation450 BeadChip arrays (n = 472). Methylation changes were verified by targeted MassARRAY analyses, assessed for their functional translation by qPCR and correlated with children’s body mass index (BMI) z scores at the age of 1 and 6 years. Further, female BALB/c mice were exposed to BPA from 1 week before mating until delivery, and weight development of their pups was monitored (n ≥ 8/group). Additionally, human adipose-derived mesenchymal stem cells were treated with BPA during the adipocyte differentiation period and assessed for exposure-related epigenetic, transcriptional and morphological changes (n = 4). Results In prenatally BPA-exposed children two CpG sites with deviating cord blood DNA-methylation profiles were identified, among them a hypo-methylated CpG in the promoter of the obesity-associated mesoderm-specific transcript (MEST). A mediator analysis suggested that prenatal BPA exposure was connected to cord blood MEST promoter methylation and MEST expression as well as BMI z scores in early infancy. This effect could be confirmed in mice in which prenatal BPA exposure altered Mest promoter methylation and transcription with a concomitant increase in the body weight of the juvenile offspring. An experimental model of in vitro differentiated human mesenchymal stem cells also revealed an epigenetically induced MEST expression and enhanced adipogenesis following BPA exposure. Conclusions Our study provides evidence that MEST mediates the impact of prenatal BPA exposure on long-term body weight development in offspring by triggering adipocyte differentiation

Novel loci for childhood body mass index and shared heritability with adult cardiometabolic traits

The genetic background of childhood body mass index (BMI), and the extent to which the well-known associations of childhood BMI with adult diseases are explained by shared genetic factors, are largely unknown. We performed a genome-wide association study meta-analysis of BMI in 61,111 children aged between 2 and 10 years. Twenty-five independent loci reached genome-wide significance in the combined discovery and replication analyses. Two of these, located near NEDD4L and SLC45A3, have not previously been reported in relation to either childhood or adult BMI. Positive genetic correlations of childhood BMI with birth weight and adult BMI, waist-to-hip ratio, diastolic blood pressure and type 2 diabetes were detected (Rg ranging from 0.11 to 0.76, P-values <0.002). A negative genetic correlation of childhood BMI with age at menarche was observed. Our results suggest that the biological processes underlying childhood BMI largely, but not completely, overlap with those underlying adult BMI. The well-known observational associations of BMI in childhood with cardio-metabolic diseases in adulthood may reflect partial genetic overlap, but in light of previous evidence, it is also likely that they are explained through phenotypic continuity of BMI from childhood into adulthood

Additional file 5: of MEST mediates the impact of prenatal bisphenol A exposure on long-term body weight development

Figure S4. Summary scheme: results overview and hypothesis indicating the influence of prenatal BPA exposure on MEST methylation and expression that is associated with adipocyte differentiation and overweight development in infant offspring. (PNG 19 kb