Expression of mRNA for phospholipase A(2), cyclooxygenases, and lipoxygenases in cultured human umbilical vascular endothelial and smooth muscle cells and in biopsies from umbilical arteries and veins

Arachidonic acid (AA) is released by phospholipase A(2) (PLA(2)) and then converted into vasoactive and inflammatory eicosanoids by cyclooxygenases (COX) and lipoxygenases (LOX). These eicosanoids are important paracrine regulators of vascular permeability, blood flow, local pro- and anticoagulant activity and they play a major role in the local inflammatory response. We have investigated the presence of mRNAs for PLA(2) and for isoforms of COX and LOX in both human endothelial cells (EC) and in human smooth muscle cells (SMC) in culture and in vascular biopsies of human umbilical veins (HUVB) and arteries (HUAB) by using the reversed transcription-polymerase chain reaction (RT-PCR) technique. Results show detectable levels of PLA(2) type IV (cPLA(2)) in cultured EC and SMC and in vascular wall biopsies from HUAB and HUVB. The cultured EC and SMC demonstrate higher levels of both COX-1 and COX-2 with PCR analyses than do vascular wall biopsies from HUAB and HUVB. This indicates a difference in the native expression of COX-1 and COX-2 in cultures of EC and SMC compared to that in biopsies from intact vessel walls. The EC and SMC in culture do not express mRNA for 5-LOX, that was, however, expressed in the vascular wall biopsies. This speaks in favour of a constitutive, i.e, in vivo expression of 5-LOX in SMC in the vascular wall of both umbilical vein and arteries. Thus results from in vitro studies of constitutive COX and LOX expression in EC and vascular SMC in culture cannot simply be extrapolated to represent in vivo conditions

Establishment of a research focus on resilient sustainable climate neutral agricultural production: resilient farming initiative

Efficient and sustainable agricultural production is a pivotal factor in meeting the nutritional needs of an expanding global population. However, it is imperative to optimize national, regional and local effectiveness to support government initiatives towards climate neutrality and resilience, while simultaneously ensuring economic viability for farmers. A significant reduction in the cost of goods must be accompanied by a decrease in their global warming potential contribution to maintain competitiveness in the world market. As such, it is necessary to adopt practices that enhance productivity while minimizing environmental impacts. This paper discusses potential solutions for the sustainable enlargement of botanical product portfolios towards essential oil products and natural extracts for value-added products, such as natural pharmaceuticals, cosmetics, agrochemicals and materials by direct waste valorization. Contributions from the fields of automation and digitalization provide the basic technology for the realization of the approaches presented. Agricultural photovoltaics can contribute to the goal of the reduction of the cost of goods and global warming potential, such as the already established utilization of biogas. The potential of the research initiative described is demonstrated by basic data on key characteristic numbers and costs from the literature. The economic potential for climate neutrality and the reduction of global warming potential contribution is seen in magnitudes of factors 5–10. A research initiative is recommended and exemplified for the industrialization of such integrated processing

XL-SIM: Extending Superresolution into Deeper Layers

Of all 3D-super resolution techniques, structured illumination microscopy (SIM) provides the best compromise with respect to resolution, signal-to-noise ratio (S/N), speed and cell viability. Its ability to achieve double resolution in all three dimensions enables resolving 3D-volumes almost 10x smaller than with a normal light microscope. Its major drawback is noise contained in the out-of-focus-signal, which-unlike the out-of-focus signal itself-cannot be removed mathematically. The resulting "noise-pollution" grows bigger the more light is removed, thus rendering thicker biological samples unsuitable for SIM. By using a slit confocal pattern, we employ optical means to suppress out-of-focus light before its noise can spoil SIM mathematics. This not only increases tissue penetration considerably, but also provides a better S/N performance and an improved confocality. The SIM pattern we employ is no line grid, but a two-dimensional hexagonal structure, which makes pattern rotation between image acquisitions obsolete and thus simplifies image acquisition and yields more robust fit parameters for SIM

Navigating Across Non-Navigable Ecore References via OCL

The Eclipse Modeling Framework (EMF) and its meta-meta model Ecore support uni-directional and bi-directional references. It is quite common that references are defined uni-directionally only because of saving storage space or separating meta models, which is problematic when implementing Object Constraint Language (OCL) constraints that require navigation against the direction of uni-directional references. This is essential for certain approaches, e.g., incremental evaluation of OCL constraints on models shown by Altenhofen et al. that is used in SAP's Modeling Infrastructure (MOIN). In this paper, we present an approach that overcomes the aforementioned issue by providing navigation across non-navigable Ecore references via OCL. We further discuss different alternative solutions and briefly describe the realization that was outcome of a project in cooperation with the SAP AG

Single Bead Labeling Method for Combining Confocal Fluorescence On-Bead Screening and Solution Validation of Tagged One-Bead One-Compound Libraries

SummaryScreening of one-bead one-compound libraries by incubating beads with fluorescently labeled target protein requires isolation and structure elucidation of a large number of primary hit beads. However, the potency of the identified ligands is only revealed after time consuming and expensive larger scale resynthesis and testing in solution. Often, many of the resynthesized compounds turn out to be weak target binders in solution due to large differences between surface and solution binding affinities. For an industry style high-throughput screening (HTS) process a high false positive rate is detrimental. We have therefore combined single bead and single molecule/single cell techniques into an integrated HTS process in which the picomole amount of substance contained on one isolated hit bead is sufficient for quality control, structure determination, and precise affinity determination to the target protein in solution

Opioid agonist treatment in transition: A cross-country comparison between Austria, Germany and Switzerland

Background and aims: OAT is a well developed and successful treatment strategy for opioid dependent patients in Europe. It has significantly contributed to the fight against the HIV and HCV pandemics, leading to an increased life expectancy in this population. Building on the OAT experiences in Austria, Germany, and Switzerland and their models of care, the objective of this study is to analyse experiences and changes in patient structures to identify necessary adaptations for the system of care. Methods: We analysed national register-based data from patients receiving OAT during the period spanning from 2010 to 2020 in Austria, Germany (cases), and Switzerland. We examined and compared OAT policies and practice at national levels through a review of literature and publicly available policy documents. Results: Across these three countries, the life expectancy of OAT patients increased substantially. The mean age increased from 33.0 in 2010 to 39.1 in 2020 in Austria, from 35.6 years to 41.5 years in Germany (cases), and from 39.6 to 47.1 in Switzerland, respectively. In all three countries, the percentage of patients/cases aged 60 years and older increased more than tenfold between 2010 and 2020. Conclusions: Integrated support models, reliable care structures, internationally comparable high treatment coverage, flexible prescribing practices, and a wide range of available OAT medications are successful strategies. The experiences in these countries indicate that it is possible to address the complex and chronic nature of opioid dependence and its concurrent mental and physical health challenges, resulting in an increasing life expectancy of OAT patients

A novel OSA-related model of intermittent hypoxia in endothelial cells under flow reveals pronounced inflammatory pathway activation

Obstructive sleep apnea (OSA) is a common sleep-related breathing disorder characterized by recurrent episodes of upper airway obstruction and subsequent hypoxia. In patients with OSA, severity and number of these hypoxic events positively correlate with the extent of associated cardiovascular pathology. The molecular mechanisms underlying intermittent hypoxia (IH)-driven cardiovascular disease in OSA, however, remain poorly understood—partly due to the lack of adequate experimental models. Here, we present a novel experimental approach that utilizes primary human endothelial cells cultivated under shear stress. Oxygen partial pressure dynamics were adopted in our in vitro model according to the desaturation-reoxygenation patterns identified in polysomnographic data of severe OSA patients (n = 10, with 892 severe desaturations, SpO2<80%). Using western blot analysis, we detected a robust activation of the two major inflammatory pathways ERK and NF-κB in endothelial cells, whereas no HIF1α and HIF2α protein stabilization was observed. In line with these findings, mRNA and protein expression of the pro-inflammatory adhesion and signaling molecule ICAM-1 and the chemokine CCL2 were significantly increased. Hence, we established a novel in vitro model for deciphering OSA-elicited effects on the vascular endothelium. First data obtained in this model point to the endothelial activation of pro-inflammatory rather than hypoxia-associated pathways in OSA. Future studies in this model might contribute to the development of targeted strategies against OSA-induced, secondary cardiovascular disease

Motor control retraining exercises for shoulder impingement: effects on function, muscle activation, and biomechanics in young adults

Objective: Evidence for effective management of shoulder impingement is limited. The present study aimed to quantify the clinical, neurophysiological, and biomechanical effects of a scapular motor control retraining for young individuals with shoulder impingement signs.Method: Sixteen adults with shoulder impingement signs (mean age 22 ? 1.6 years) underwent the intervention and 16 healthy participants (24.8 ? 3.1years) provided reference data. Shoulder function and pain were assessed using the Shoulder Pain and Disability Index (SPADI) and other questionnaires. Electromyography (EMG) and 3 dimensional motion analysis was used to record muscle activation and kinematic data during arm elevation to 90? and lowering in 3 planes. Patients were assessed pre and post a 10-week motor control based intervention, utilizing scapular orientation retraining.Results: Pre-intervention, patients reported pain and reduced function compared to the healthy participants (SPADI in patients 20 ? 9.2; healthy 0 ? 0). Post intervention, the SPADI scores reduced significantly (P < .001) by a mean of 10 points (?4). EMG showed delayed onset and early termination of serratus anterior and lower trapezius muscle activity pre-intervention, which improved significantly post-intervention (P < .05). Pre intervention, patients exhibited on average 4.6-7.4? less posterior tilt, which was significantly lower in 2 arm elevation planes (P < .05) than healthy participants. Postintervention, upward rotation and posterior tilt increased significantly (P <.05) during 2 arm movements, approaching the healthy values.Conclusion: A 10-week motor control intervention for shoulder impingement increased function and reduced pain. Recovery mechanisms were indicated by changes in muscle recruitment andscapular kinematics. The efficacy of the intervention requires further examined in a randomizedcontrol trial

Neutrophils promote venular thrombosis by shaping the rheological environment for platelet aggregation

In advanced inflammatory disease, microvascular thrombosis leads to the interruption of blood supply and provokes ischemic tissue injury. Recently, intravascularly adherent leukocytes have been reported to shape the blood flow in their immediate vascular environment. Whether these rheological effects are relevant for microvascular thrombogenesis remains elusive. Employing multi-channel in vivo microscopy, analyses in microfluidic devices, and computational modeling, we identified a previously unanticipated role of leukocytes for microvascular clot formation in inflamed tissue. For this purpose, neutrophils adhere at distinct sites in the microvasculature where these immune cells effectively promote thrombosis by shaping the rheological environment for platelet aggregation. In contrast to larger (lower-shear) vessels, this process in high-shear microvessels does not require fibrin generation or extracellular trap formation, but involves GPIb alpha-vWF and CD40-CD40L-dependent platelet interactions. Conversely, interference with these cellular interactions substantially compromises microvascular clotting. Thus, leukocytes shape the rheological environment in the inflamed venular microvasculature for platelet aggregation thereby effectively promoting the formation of blood clots. Targeting this specific crosstalk between the immune system and the hemostatic system might be instrumental for the prevention and treatment of microvascular thromboembolic pathologies, which are inaccessible to invasive revascularization strategies

Complexity in a systems engineering organization: An empirical case study

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.ystems Engineering development projects often fail to meet delivery expectations in terms of timescales and cost. Project plans, which set cost and deadline expectations, are produced and monitored within a reductionist paradigm, incorporating a deterministic view of cause and effect. This assumes that the cumulative activities and their corresponding durations that comprise the developed solution can be known in advance, and that monitoring and management intervention can ensure satisfactory delivery of an adequate solution, through implementation of this plan. This paper presents a case study that examines the Systems Engineering function within a Thales UK business line. The focus is the organization. The research is exploratory. It gathers evidence through participant-observation, interviews, documentation, and archival records. It considers two perspectives, a ‘traditional’ predominantly reductionist perspective, and a novel Complex Adaptive System (CAS) perspective. Evidence is analyzed in light of both perspectives to consider how each is able to explain the observations. Research that considers an organization as a CAS is predominately theoretical, rather than empirical. This paper contributes by viewing a systems engineering development organisation as a CAS, and considering the novel insights this perspective brings to the issue of satisfactory project delivery