Automated universal chip platform for fluorescence based cellular assays

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.The advantage of cell based assays used as biosensors is the direct access to hardly obtainable parameters like toxicity, mutagenicity and pharmacological effectiveness. Within the last few years we established a micro fluidic platform including a peristaltic micro pump as well as several valves, manifolds and micro channels [1]. For optical online monitoring the micro fluidic system is bonded to a glass slide. Furthermore the biochip is fixed on an electrically heated support. The pneumatically actuated peristaltic pump as well as the temperature control is performed by a control device. For the fluorescence based online monitoring a robotic guided fluorescence measurement module was developed, which supports the detection of fluorescence in microtiter plates and microfluidic systems. This measurement module allows the fluorescence detection of two different excitation / detection wavelengths (480 / 530 nm and 570 / 620 nm) and was successfully characterised using EGFP and Rhodamine 6G. Additionally three cell based assays with bacterial, yeast and human cells were characterized

Soils potentially vulnerable to phosphorus losses: speciation of inorganic and organic phosphorus and estimation of leaching losses

Eutrophication is an important threat to aquatic ecosystems world-wide, and reliable identification of areas vulnerable to phosphorus (P) losses from diffuse sources is essential for high efficiency of mitigation measures. In this three-step study we investigated (i) relationships between the agronomic (Olsen-P and P-AL) and environmental soil P tests (P-CaCl2) with molecular techniques (P-31 NMR and XANES) followed by (ii) rainfall simulation experiment on topsoil lysimeters and (iii) comparison to long-term field measurements of water quality. Soil samples were collected from seven sites indicated to be vulnerable to nutrient losses due to underlying geology. High P release correlated to standard agronomic P tests (Olsen P, r = 0.67; and P-AL, r = 0.74) and low P sorption capacity (r = - 0.5). High content of iron-bound P compounds indicated more labile P and higher release of dissolved P (r = 0.67). The leaching experiment showed that three out of four soils with high initial soil P status had both higher P leaching concentrations before fertilization (0.83-7.7 mg P l(-1)) compared to soil with low initial soil P status (0.007-0.23 mg P l(-1)), and higher increase in P concentrations after fertilization. Higher soil P sorption capacity reduced P leaching losses. Finally, long-term monitoring data show no significant trends in P losses in a field with low initial P content and moderate P fertilization rates whereas high and over time increasing P losses were recorded in a field with high initial soil P content and repetitively high P fertilization rates

A modular in vitro flow model to analyse blood-surface interactions under physiological conditions

Newly developed materials for blood-contacting devices need to undergo hemocompatibility testing to prove compliance with clinical requirements. However, many current in vitro models disregard the influence of flow conditions and blood exchange as it occurs in vivo. Here, we present a flow model which allows testing of blood-surface interactions under more physiological conditions. This modular platform consists of a triple-pump-chip and a microchannel-chip with a customizable surface. Flow conditions can be adjusted individually within the physiological range. A performance test with whole blood confirmed the hemocompatibility of our modular platform. Hemolysis was negligible, inflammation and hemostasis parameters were comparable to those detected in a previously established quasi-static whole blood screening chamber. The steady supply of fresh blood avoids secondary effects by nonphysiological accumulation of activation products. Experiments with three subsequently tested biomaterials showed results similar to literature and our own experience. The reported results suggest that our developed flow model allows the evaluation of blood-contacting materials under physiological flow conditions. By adjusting the occurring wall shear stress, the model can be adapted for selected test conditions

The effect of sacubitril/valsartan compared to olmesartan on cardiovascular remodelling in subjects with essential hypertension: the results of a randomized, double-blind, active-controlled study

Aims: Progressive aortic stiffening eventually leads to left ventricular (LV) hypertrophy and heart failure if left untreated. Anti-hypertensive agents have been shown to reverse this to some extent. The effects of sacubitril/valsartan (LCZ696), a dual-action angiotensin receptor blocker (ARB), and neprilysin inhibitor, on arterial stiffness and LV remodelling have not been investigated. Methods and results: This was a randomized, multi-centre, double-blind, double-dummy, active-controlled, parallel group, study to compare the effects on cardiovascular remodelling of sacubitril/valsartan with those of olmesartan in patients with hypertension and elevated pulse pressure. Magnetic resonance imaging scans were used to assess LV mass and local aortic distensibility, at baseline and at 12 and 52 weeks after initiation of treatment. Central pulse and systolic pressure were determined using a SphymoCor® XCEL device at each time point. A total of 114 patients were included, with 57 in each treatment group. The mean age was 59.8 years, and 67.5% were male. Demographic characteristics did not vary between the two sets of patients. Left ventricular mass index decreased to a greater extent in the sacubitril/valsartan group compared to the olmesartan group from baseline to 12 weeks (−6.36 vs. −2.32 g/m2; P = 0.039) and from baseline to 52 weeks (−6.83 vs. −3.55 g/m2; P = 0.029). These differences remained significant after adjustment for systolic blood pressure (SBP) at follow-up (P = 0.036 and 0.019 at 12 and 52 weeks, respectively) and similar signals (though formally non-significant) were observed after adjusting for changes in SBP (P = 0.0612 and P = 0.0529, respectively). There were no significant differences in local distensibility changes from baseline to 12 or 52 weeks between the two groups; however, there was a larger reduction in central pulse pressure for the sacubitril/valsartan group compared to the olmesartan group (P = 0.010). Conclusion: Since LV mass change correlates with cardiovascular prognosis, the greater reductions in LV mass indicate valuable advantages of sacubitril/valsartan compared to olmesartan. The finding that LV mass index decrease might be to some extent independent of SBP suggests that the effect of the dual-acting agent may go beyond those due to its BP-lowering ability

Automated substance testing for lab-on-chip devices : From 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better Health Lille, France. 23-26 June 2013

First published by BioMed Central: Kloke, Lutz ; Schimek, Katharina ; Brincker, Sven ; Lorenz, Alexandra ; Jänicke, Annika ; Drewell, Christopher ; Hoffmann, Silke ; Busek, Mathias ; Sonntag, Frank ; Danz, Norbert ; Polk, Christoph ; Schmieder, Florian ; Borchanikov, Alexey ; Artyushenko, Viacheslav ; Baudisch, Frank ; Bürger, Mario ; Horland, Reyk ; Lauster, Roland ; Marx, Uwe : Automated substance testing for lab-on-chip devices : From 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better Health Lille, France. 23-26 June 2013. - In: BMC Proceedings. - ISSN 1753-6561 (online). - 7 (2013), suppl. 6, P28. - doi:10.1186/1753-6561-7-S6-P28

The genomes of two key bumblebee species with primitive eusocial organization

Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats. Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits. Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation

Определение природных и техногенных радионуклидов в бальнеологических объектах

Quantitative detection of angiogenic biomarkers provides a powerful tool to diagnose cancers in early stages and to follow its progression during therapy. Conventional tests require trained personnel, dedicated laboratory equipment and are generally time-consuming. Herein, we propose our developed biosensing platform as a useful tool for a rapid determination of Angiopoietin-2 biomarker directly from patient plasma within 30 minutes, without any sample preparation or dilution. Bloch surface waves supported by one dimensional photonic crystal are exploited to enhance and redirect the fluorescence arising from a sandwich immunoassay that involves Angiopoietin-2. The sensing units consist of disposable and low-cost plastic biochips coated with the photonic crystal. The biosensing platform is demonstrated to detect Angiopoietin-2 in plasma samples at the clinically relevant concentration of 6 ng/mL, with an estimated limit of detection of approximately 1 ng/mL. This is the first Bloch surface wave based assay capable of detecting relevant concentrations of an angiogenic factor in plasma samples. The results obtained by the developed biosensing platform are in close agreement with enzyme-linked immunosorbent assays, demonstrating a good accuracy, and their repeatability showed acceptable relative variations

Globally distributed iridium layer preserved within the Chicxulub impact structure

The Cretaceous-Paleogene (K-Pg) mass extinction is marked globally by elevated concentrations of iridium, emplaced by a hypervelocity impact event 66 million years ago. Here, we report new data from four independent laboratories that reveal a positive iridium anomaly within the peak-ring sequence of the Chicxulub impact structure, in drill core recovered by IODP-ICDP Expedition 364. The highest concentration of ultrafine meteoritic matter occurs in the post-impact sediments that cover the crater peak ring, just below the lowermost Danian pelagic limestone. Within years to decades after the impact event, this part of the Chicxulub impact basin returned to a relatively low-energy depositional environment, recording in unprecedented detail the recovery of life during the succeeding millennia. The iridium layer provides a key temporal horizon precisely linking Chicxulub to K-Pg boundary sections worldwide