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Merkel, Heinze, Hilbert, and Naegele focus on the benefits that modern technology offers for active ageing. The chapter concentrates on three different areas: information and communication technologies (ICT), housing, and mobility. The authors analyse current developments and trends in each of those areas and also focus on challenges. They argue that despite its potential, the adoption, implementation, and diffusion of innovative technologies are behind expectations. Europe is presently facing a deployment gap: research and development efforts on the one hand and only limited market success on the other. The authors explore the reasons for the deployment gap and, moreover, provide examples of good practice and conclude with recommendations for further research as well as implications for policy and practice.

In mammals, the differentiation and maturation of megakaryocytes (MKs) occurs in the bone marrow (BM). The three-dimensional environment influences megakaryopoiesis and platelet release. Thus, imaging MKs within the intact BM is important to understand megakaryopoiesis. Here, we present an optical clearing protocol for intact bones and the subsequent microscopic analysis including image processing to quantitatively assess MK size and distribution. This technique overcomes the limitations of classical sectioning methods as the entire bone can be imaged.

Diagnosen zur gegenwartigen Gesellschaftsformation beschreiben in letzter Zeit eine wachsende Dynamik soziookonomischer Verunsicherungen; allgemein werden oft grundlegende soziale Umbruche konstatiert. Generell beschreiben Konzepte sozialen Wandels Veranderungen der sozialen Strukturen einer Gesellschaft in ihren Basisinstitutionen, Kulturen, subjektiven Deutungsmustern und Lebensformen. Vornehmlich zielt er auf die Veranderung von Familien- und Haushaltsstrukturen, aber auch auf dem Arbeitsmarkt oder hinsichtlich von Wertevorstellungen ab. Richtet man den Blick auf Gesellschaftsdiagnosen, wird immer mehr von einer „Abstiegsgesellschaft“ gesprochen sowie vom „erschopften“ Selbst als gesamtgesellschaftliches Phanomen.

Wilms tumor (WT) is the most common kidney cancer in childhood. Mutations in the microprocessor genes DROSHA and DGCR8 have been identified as putative oncogenic drivers, indicating a critical role of aberrant miRNA processing in WT formation. To characterize the in vivo role of DROSHA mutations during kidney development and their oncogenic potential, we analyzed mouse lines with either a targeted deletion of Drosha or an inducible expression of human DROSHA carrying a tumor-specific E1147K mutation that acts in a dominant negative manner. Both types of mutation induce striking changes in miRNA patterns. Six2-cre mediated deletion of Drosha in nephron progenitors led to perinatal lethality with apoptotic loss of progenitor cells and early termination of nephrogenesis. Mosaic deletions via Wt1-cre

Cation ordering is expected to influence the properties of oxides in the $\mathrm{Zr}{\mathrm{O}}_{2}\text{\ensuremath{-}}\mathrm{YTa}{\mathrm{O}}_{4}$ system, but the ordering patterns and their relative energies are not well understood. This work investigates the stability of zirconium-yttrium-tantalum orderings over the parent $t\text{\ensuremath{-}}\mathrm{Zr}{\mathrm{O}}_{2}$ and $m\text{\ensuremath{-}}\mathrm{Zr}{\mathrm{O}}_{2}$ crystal structures from first principles. The calculations predict a strong tendency for yttrium and tantalum atoms to cluster into checkerboard layers throughout the entire quasibinary. Such checkerboard layers are in fact found to be the building blocks for the lowest-energy $\mathrm{YTa}{\mathrm{O}}_{4}$ structures, e.g., ${M}^{\ensuremath{'}}$, $M$, and $T\text{\ensuremath{-}}\mathrm{YTa}{\mathrm{O}}_{4}$. The stability of these orderings is related to the propensity of each cation to achieve its favored oxygen coordination. The preference to cluster in the alloyed materials suggests a tendency for short-range ordering. It is hypothesized that checkerboard-type short-range order is a global trend in all $\mathrm{Zr}{\mathrm{O}}_{2}$ systems with charge-compensating stabilizers, notably rare-earth tantalates and niobates, and has important implications for ferroelastic toughening mechanisms in these materials. A study of the transformation pathways connecting $M$ and $T\text{\ensuremath{-}}\mathrm{YTa}{\mathrm{O}}_{4}$ predicts that unlike $t\text{\ensuremath{-}}\mathrm{Zr}{\mathrm{O}}_{2},T\text{\ensuremath{-}}\mathrm{YTa}{\mathrm{O}}_{4}$ is dynamically unstable, suggesting that anharmonic vibrations stabilize this phase and must be introduced to rigorously account for temperature effects.

The regulation of immune cell migration throughout the body is essential to warrant immunosurveillance and to maintain immune homeostasis. Marking and tracking of these cells has proven important to study mechanisms of immune cell trafficking and cell interaction in vivo. Photoconversion is a well-suited technique for intravital application because it enables contactless time- and location-specific marking of cells in the tissue without surgically manipulating the microenvironment of the cells in question. However, in dividing cells the converted fluorescent protein may decline quickly. Here, we provide a detailed description of the photoconversion technique and its applicability to tracking highly proliferating T cells from the priming site of T cell activation to peripheral target organs of effector function in a preclinical model. Dendra2+ T cells were photoconverted in the Peyer’s patches during the initiation phase of acute graft-versus-host disease (GvHD) and tracked through the mesenteric lymph nodes and the peripheral blood to the small intestine with flow cytometry and intravital two-photon microscopy. Photoconverted alloreactive T cells preserved the full proliferative capacity, homing, and migration of alloreactive T cells in the intestinal lamina propria. We conclusively proved that photoconversion of highly proliferative alloreactive T cells in the Peyer’s patches is an effective tool to study trafficking of alloreactive T cells under physiologic conditions and to GvHD target tissues. This technique can also be applied to the study of immune cell tracking under inflammatory and non-inflammatory conditions.

Aims\ud To assess the prevalence and management of depressive disorders in people with Type 2 diabetes in different countries.\ud \ud Methods\ud People with diabetes aged 18–65 years and treated in outpatient settings were recruited in 14 countries and underwent a psychiatric interview. Participants completed the Patient Health Questionnaire and the Problem Areas in Diabetes scale. Demographic and medical record data were collected.\ud \ud Results\ud A total of 2783 people with Type 2 diabetes (45.3% men, mean duration of diabetes 8.8 years) participated. Overall, 10.6% were diagnosed with current major depressive disorder and 17.0% reported moderate to severe levels of depressive symptomatology (Patient Health Questionnaire scores >9). Multivariable analyses showed that, after controlling for country, current major depressive disorder was significantly associated with gender (women) (PPPPPConclusions\ud Our international study, the largest of this type ever undertaken, shows that people with diabetes frequently have depressive disorders and also significant levels of depressive symptoms. Our findings indicate that the identification and appropriate care for psychological and psychiatric problems is not the norm and suggest a lack of the comprehensive approach to diabetes management that is needed to improve clinical outcomes.

Background— E193, a heterozygous truncating mutation in the human transcription cofactor Eyes absent 4 (Eya4), causes hearing impairment followed by dilative cardiomyopathy. Methods and Results— In this study, we first show Eya4 and E193 alter the expression of p27 kip1 in vitro, suggesting Eya4 is a negative regulator of p27. Next, we generated transgenic mice with cardiac-specific overexpression of Eya4 or E193. Luciferase and chromatin immunoprecipitation assays confirmed Eya4 and E193 bind and regulate p27 expression in a contradictory manner. Activity and phosphorylation status of the downstream molecules casein kinase-2α and histone deacetylase 2 were significantly elevated in Eya4- but significantly reduced in E193-overexpressing animals compared with wild-type littermates. Magnetic resonance imaging and hemodynamic analysis indicate Eya4-overexpression results in an age-dependent development of hypertrophy already under baseline conditions with no obvious functional effects, whereas E193 animals develop onset of dilative cardiomyopathy as seen in human E193 patients. Both cardiac phenotypes were aggravated on pressure overload. Finally, we identified a new heterozygous truncating Eya4 mutation, E215, which leads to similar clinical features of disease and a stable myocardial expression of the mutant protein as seen with E193. Conclusions— Our results implicate Eya4/Six1 regulates normal cardiac function via p27/casein kinase-2α/histone deacetylase 2 and indicate that mutations within this transcriptional complex and signaling cascade lead to the development of cardiomyopathy.

Colon cancer is one of the most common tumors in the human population. Recent studies have shown a reduced risk for colon cancer in patients given the antidepressant fluoxetine (FLX). The exact mechanism by which FLX might protect from colon cancer remains however controversial. Here, FLX reduced the development of different colon tumor xenografts, as well as proliferation in hypoxic tumor areas within them. FLX treatment also decreased microvessel numbers in tumors. Although FLX did not increase serum and tumor glucose levels as much as the colon chemotherapy gold standard Fluorouracil did, lactate levels were significantly augmented within tumors by FLX treatment. The gene expression of the MCT4 lactate transporter was significantly downregulated. Total protein amounts from the third and fifth mitochondrial complexes were significantly decreased by FLX in tumors. Cell culture experiments revealed that FLX reduced the mitochondrial membrane potential significantly and disabled the reactive oxygen species production of the third mitochondrial complex. Furthermore, FLX arrested hypoxic colon tumor cells in the G0/G1 phase of the cell-cycle. The expression of key cell-cycle-related checkpoint proteins was enhanced in cell culture and in vivo experiments. Therefore, we suggest FLX impairs energy generation, cell cycle progression and proliferation in tumor cells, especially under condition of hypoxia. This then leads to reduced microvessel formation and tumor shrinkage in xenograft models.

In mammals, the differentiation and maturation of megakaryocytes (MKs) occurs in the bone marrow (BM). The three-dimensional environment influences megakaryopoiesis and platelet release. Thus, imaging MKs within the intact BM is important to understand megakaryopoiesis. Here, we present an optical clearing protocol for intact bones and the subsequent microscopic analysis including image processing to quantitatively assess MK size and distribution. This technique overcomes the limitations of classical sectioning methods as the entire bone can be imaged.

Der Begriff der sozialen Innovation ist zwar noch relativ neu, hat allerdings in den letzten Jahren Karriere gemacht und wird in Zielvorstellungen der EU an zentraler Stelle genannt. Dabei wird ein Innovationsbegriff verwandt, der uber naturwissenschaftlich-technische Produkt- oder Marktinnovationen hinausgeht und die Neukonfiguration sozialer Arrangements mit einbezieht. Somit wird der Blick auf heterogene Akteure, Interdisziplinaritat und Reflexivitat gerichtet.

Der Beitrag konzentriert sich auf Techniken, die das selbststandige Wohnen im Alter unterstutzen, mogliche altersbedingte Einschrankungen und damit die Lebensqualitat Alterer positiv beeinflussen konnen. Das Spektrum reicht von Hausnotrufsystemen bis hin zur Telemedizin. Obwohl gerade das Alter pradestiniert fur den Einsatz flexibel unterstutzender Technologien ist, werden in Deutschland jedoch Implementationsprobleme sichtbar.

Total internal reflection fluorescence microscopy (TIRF-M) provides low-invasive high-contrast surface imaging with optical sectioning of typically 100–200 nm. Thus, TIRF-M has become an established tool for imaging surfaces, including cell membranes. For TIRF-M, a homogenous evanescent field of excitation over the whole field of view is generally desired for quantitative microscopy; however, this is not necessarily straightforward to generate with Gaussian beams. In recent years, several improvements on TIRF-M have been developed that have addressed non-uniform scattering fringes and other artifacts. Here, we introduce a cost-effective TIRF setup with a very low degree of complexity and no moving parts, which provides a flattop-like excitation profile. The setup uses a tunable laser ring zoom focus system to generate a full 360° TIRF illumination. Two axicon lenses and one focus lens allow for generation and control of the ring diameter to tune the TIRF excitation angle. We show that 360° laser illumination in combination with a radial polarizer will generate an evanescent Bessel-beam excitation field that exhibits a flattop intensity over an extended part of the field of view, and demonstrate the advantages of this axicon-based Bessel beam illumination for live-cell imaging.

In continuously beating cells like cardiac myocytes, there are rapid alterations of cytosolic Ca

Super-resolution techniques such as direct Stochastic Optical Reconstruction Microscopy (dSTORM) have become versatile and well-established tools for biological imaging over the last century. Here, we theoretically and experimentally show that clever combination of different fluorescence modalities allows further improvements. We found that the interaction of fluorophores with plasmonic surfaces boost super-resolution performance in dSTORM approaches as it allows for tailoring the excitation and emission properties. The strength of the approach is that no further specialized microscope setup is required as the described enhancement solely rely on metal-dielectric coated glass coverslips that are straightforward to fabricate. Such biocompatible plasmonic nanolayers enhance the signal-to-noise ratio of dSTORM, and thus sharpens the localization precision by a factor of two.

This deposit is composed by a publication in which the IGC's authors have had the role of collaboration (it's a collaboration publication). This type of deposit in ARCA is in restrictedAccess (it can't be in open access to the public), and can only be accessed by two ways: either by requesting a legal copy from the author (the email contact present in this deposit) or by visiting the following link: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5373357/ This publication hasn't any creative commons license associated. In recent years, the myocardium has been rediscovered under the lenses of immunology, and lymphocytes have been implicated in the pathogenesis of cardiomyopathies with different etiologies. Aging is an important risk factor for heart diseases, and it also has impact on the immune system. Thus, we sought to determine whether immunological activity would influence myocardial structure and function in elderly mice. Morphological, functional, and molecular analyses revealed that the age-related myocardial impairment occurs in parallel with shifts in the composition of tissue-resident leukocytes and with an accumulation of activated CD4+Foxp3-(forkhead box P3) IFN-γ+T cells in the heart-draining lymph nodes. A comprehensive characterization of different aged immune-deficient mouse strains revealed that T cells significantly contribute to age-related myocardial inflammation and functional decline. Upon adoptive cell transfer, the T cells isolated from the mediastinal lymph node (med-LN) of aged animals exhibited increased cardiotropism, compared with cells purified from young donors or from other irrelevant sites. Nevertheless, these cells caused rather mild effects on cardiac functionality, indicating that myocardial aging might stem from a combination of intrinsic and extrinsic (immunological) factors. Taken together, the data herein presented indicate that heart-directed immune responses may spontaneously arise in the elderly, even in the absence of a clear tissue damage or concomitant infection. These observations might shed new light on the emerging role of T cells in myocardial diseases, which primarily affect the elderly population. Bundesministerium für Bildung und Forschung grants: (BMBF01, EO1004); German Research Foundation grant: (DFG SFB688, TP A10, and B07); Brazilian National Council for Scientific and Technological Development. info:eu-repo/semantics/publishedVersion

The definition of sub-20 nm electronic devices for the newest generation of smart phones, computer and automotive is calling for very innovative FEOL wet chemical cleans. The electronic properties are very sensitive, in respect of the surface morphology on a Si-wafer. Most of the modern wet cleans are based on the RCA-clean [1]. Innovative cleans, like for example the IMEC-clean [2] and modified RCA clean were developed, using ozone-DIW mixture (O3-DIW), in order to improve the cleaning performance [3], [4]. Since several years electrolyzed water (EW) is used in semiconductor manufacturing [5]. An electrochemical reaction is induced by an electrode and a small amount of ammonia hydroxide (NH4OH) or ammonia sulfate and DIW [7].

Ethnopharmacological relevance Agastache mexicana subspecies mexicana (Amm) and xolocotziana (Amx) are used in Mexican traditional medicine to relief cultural affiliation syndromes known as “susto” or “espanto”, for “nervous” condition, and as a sleep aid. Despite its intensive use, neuropharmacological studies are scarce, and the chemical composition of the aqueous extracts has not been described. Aims of the study are: (1) To analyze the chemical composition of aqueous extracts from aerial parts of Amm and Amx. (2) To evaluate the anxiolytic-like, sedative, antidepressant-like effects. (3) Analyze the general toxic effects of different doses. Materials and methods Anxiolytic-like and sedative effects were measured in the avoidance exploratory behavior, burying behavior and the hole-board tests. The antidepressant-like actions were studied in the forced swimming and tail suspension tests. Finally, general activity and motor coordination disturbances were evaluated in the open field, inverted screen and rota-rod tests. The acute toxicity of Amm and Amx was determined by calculating their LD 50 (mean lethal dose). The chemical analyses were performed employing chromatographic, photometric and HPLC–ESI-MS techniques. Results Low doses of Amm and Amx (0.1σ1.0 mg/kg) induced anxiolytic-like actions; while higher doses (over 10 mg/kg) induced sedation and reduced the locomotor activity, exerting a general inhibition in the central nervous system (CNS). Conclusions Results support the use of Amm and Amx in traditional medicine as tranquilizers and sleep inducers. Additionally, this paper contributes to the knowledge of the chemical composition of the aqueous extracts of these plants.

This contribution discusses the hopes associated with the rise of the creative industries and gives explanations for the debates in politics, science and the media. In doing so, our underlying thesis is that the culture and creative economy is a virtual sector and that a uniform promotion—also by means of staged events that attract a lot of media attention—needs to be challenged. In a further step, the scientific implications will also be outlined on the basis of the term's “career” in the public debate. A brief analysis of the empiricism of the creative economy will provide further insight into its real significance and demonstrate possible definitional weaknesses of the term. The creative industries are an essential element of modern economic infrastructure. They will play an important role in the future, especially for cluster strategies. However, the scientific research so far is not able to reach more accurate conclusions regarding the effects of governance on the culture and creative industry. There...

Regulated reorganization of the actin cytoskeleton is a prerequisite for proper platelet production and function. Consequently, defects in proteins controlling actin dynamics have been associated with platelet disorders in humans and mice. Twinfilin 2a (Twf2a) is a small actin-binding protein that inhibits actin filament assembly by sequestering actin monomers and capping filament barbed ends. Moreover, Twf2a binds heterodimeric capping proteins, but the role of this interaction in cytoskeletal dynamics has remained elusive. Even though Twf2a has pronounced effects on actin dynamics in vitro, only little is known about its function in vivo. Here, we report that constitutive Twf2a-deficient mice (Twf2a-/-) display mild macrothrombocytopenia due to a markedly accelerated platelet clearance in the spleen. Twf2a-/- platelets showed enhanced integrin activation and α-granule release in response to stimulation of (hem) immunoreceptor tyrosine-based activation motif (ITAM) and G-protein-coupled receptors, increased adhesion and aggregate formation on collagen I under flow, and accelerated clot retraction and spreading on fibrinogen. In vivo, Twf2a deficiency resulted in shortened tail bleeding times and faster occlusive arterial thrombus formation. The hyperreactivity of Twf2a-/- platelets was attributed to enhanced actin dynamics, characterized by an increased activity of n-cofilin and profilin 1, leading to a thickened cortical cytoskeleton and hence sustained integrin activation by limiting calpain-mediated integrin inactivation. In summary, our results reveal the first in vivo functions of mammalian Twf2a and demonstrate that Twf2a-controlled actin rearrangements dampen platelet activation responses in a n-cofilin- and profilin 1-dependent manner, thereby indirectly regulating platelet reactivity and half-life in mice.

Gesundheitsbezogene Branchen und Gestaltungsfelder haben in den letzten Jahren den wirtschaft lichen Strukturwandel mit gepragt und waren verantwortlich fur die Schaffung vieler Arbeitsplatze im Ruhrgebiet. Allein zwischen 2008 und 2013 entstanden mehr als 32.000 zusatzliche Arbeitsplatze. Die Spannbreite reicht von der ambulanten und stationaren Versorgung (dem traditionellen Gesundheitswesen) uber die Medizintechnik und die Gesundheitshandwerke bis hin zum Service Wohnen fur pflegebedurftige Menschen oder der Medical Wellness.

In mammals, megakaryocytes (MKs) in the bone marrow (BM) produce blood platelets, required for hemostasis and thrombosis. MKs originate from hematopoietic stem cells and are thought to migrate from an endosteal niche towards the vascular sinusoids during their maturation. Through imaging of MKs in the intact BM, here we show that MKs can be found within the entire BM, without a bias towards bone-distant regions. By combining in vivo two-photon microscopy and in situ light-sheet fluorescence microscopy with computational simulations, we reveal surprisingly slow MK migration, limited intervascular space, and a vessel-biased MK pool. These data challenge the current thrombopoiesis model of MK migration and support a modified model, where MKs at sinusoids are replenished by sinusoidal precursors rather than cells from a distant periostic niche. As MKs do not need to migrate to reach the vessel, therapies to increase MK numbers might be sufficient to raise platelet counts., Megakaryocyte maturation is thought to occur as the cells migrate from a vessel-distant (endosteal) niche to the vessel within the bone. Here, the authors show that megakaryocytes represent largely sessile cells in close contact with the vasculature and homogeneously distributed in the bone marrow.

Humans are continuously exposed to airborne spores of the saprophytic fungus Aspergillus fumigatus. However, in healthy individuals pulmonary host defense mechanisms efficiently eliminate the fungus. In contrast, A. fumigatus causes devastating infections in immunocompromised patients. Host immune responses against A. fumigatus lung infections in immunocompromised conditions have remained largely elusive. Given the dynamic changes in immune cell subsets within tissues upon immunosuppressive therapy, we dissected the spatiotemporal pulmonary immune response after A. fumigatus infection to reveal basic immunological events that fail to effectively control invasive fungal disease. In different immunocompromised murine models, myeloid, notably neutrophils, and macrophages, but not lymphoid cells were strongly recruited to the lungs upon infection. Other myeloid cells, particularly dendritic cells and monocytes, were only recruited to lungs of corticosteroid treated mice, which developed a strong pulmonary inflammation after infection. Lymphoid cells, particularly CD4\(^+\) or CD8\(^+\) T-cells and NK cells were highly reduced upon immunosuppression and not recruited after A. fumigatus infection. Moreover, adoptive CD11b\(^+\) myeloid cell transfer rescued cyclophosphamide immunosuppressed mice from lethal A. fumigatus infection but not cortisone and cyclophosphamide immunosuppressed mice. Our findings illustrate that CD11b\(^+\) myeloid cells are critical for anti-A. fumigatus defense under cyclophosphamide immunosuppressed conditions.

Ethnopharmacological relevance Dracocephalum moldavica is used as a tranquilizer and as remedy for nervous conditions relief in the Mexican traditional medicine. Despite its intensive use no literature reported neuropharmacological studies on Dracocephalum moldavica as yet. Aim of the study The sedative, anxiolytic-like and antidepressant-like effects of the aqueous extract of aerial parts of Dracocephalum moldavica (Lamiaceae) (DM) were evaluated in behavioral models in mice. The general toxic effects of DM were evaluated as well as their chemical analysis was performed. Materials and methods DM effects were evaluated on pentobarbital-induced sleeping time (SPT), the hole-board (HBT), and the avoidance exploratory behavior (AEBT) tests and on the forced swimming test (FST). General activity and motor coordination were evaluated in the open field (OFT) and Rota-rod tests, respectively. The acute toxicity of DM was determinate by its LD50 dose. The chemical analyses DM were performed by chromatographic and HPLC–ESI-MS techniques. Results DM prolonged the pentobarbital-induced sleeping time, induced sedation in the HBT, decreased spontaneous activity and produced motor coordination impairment in mice. However, DM did not show anxiolytic effects in the AEBT or HBT and it was not effective in FST. The DM-treatment produced mortalities with LD50 = 470 mg/kg body weight. The HPLC–ESI-MS analysis of DM revealed that (acacetin, apigenin and luteolin)-7-O-β- d -(6″-O-malonyl)-glucoside derivates are the main compounds of DM. Conclusions DM induced sedative actions and a general inhibition of CNS activity observed by the decrease of animals’ general activity, motor coordination and exploration.

Blood platelets play key roles in hemostasis and thrombosis and are the second most abundant cell type in the circulation. Due to their short life span of only a few days, anuclear platelets are continuously replenished and thus provide a classic system to study hematopoiesis. In mammals, platelets are produced by megakaryocytes (MKs) that are predominantly residing in the bone marrow (BM). MKs originate from hematopoietic stem cells and are thought to migrate from an endosteal niche towards the vascular sinusoids during their maturation. Unfortunately, previous studies on megakaryopoiesis were often limited by 2D imaging and cutting artefacts when analyzing bone sections, potentially resulting in underestimation of MK-to-vessel contacts and MK volumes. We studied megakaryopoiesis by visualizing MKs in their 3D environment. To this end, murine bones were simultaneously stained for MKs and endothelial cells, fixed, chemically cleared and imaged by Light Sheet Fluorescence Microscopy (LSFM). Thus, we achieved 3D-reconstructions of the complete and intact bone with subcellular resolution. Through imaging of MKs in the intact BM, we show that MKs can be found within the entire BM, without a bias towards bone-distant regions. We developed and compared different image processing pipelines and simulation scenarios for precise identification of MKs in 3D light-sheet fluorescence microscopy of uncut murine bones. By combining in vivo two-photon microscopy and in situ LSFM with computational simulations, we reveal surprisingly slow MK migration, limited intervascular space, and a vessel-biased MK pool. To complement limited imaging approaches computational simulations represent an important, well-controllable tool. Typically, simulation studies use artificial meshes as templates to minimize the computational effort or due to the lack of experimental data. Unfortunately, such simplified artificial templates for MKs and the vasculature can bias simulations and lead to misinterpretations as we show here. Using the segmented cell and vessel objects of true 3D images can overcome those limitations providing a simulation framework that has the prerequisites to maximally reflect the physiological situation. Thus, imaging and simulations go hand in hand when the respective 3D cell and vessel objects perfectly serve as biological templates for advanced simulations. We demonstrate reliable whole-bone analysis in silico, and found that MKs influence neutrophil and HSC migration as biomechanical restrainers modulating cell mobility and extravasation. These data challenge the current thrombopoiesis model of MK migration and support a modified model, where MKs at sinusoids are replenished by sinusoidal precursors rather than cells from a distant periostic niche (1). Furthermore, we identify MKs as biomechanical restraints for bone marrow cell mobilization. As MKs themselves do not need to migrate to reach the vessel, therapies to increase MK numbers might be sufficient to raise platelet counts. (1) Stegner D, van Eeuwijk JMM, Angay O, Gorelashvili MG, Semeniak D, Pinnecker J, Schmithausen P, Meyer I, Friedrich M, Dütting S, Brede C, Beilhack A, Schulze H, Nieswandt B, Heinze KG. Thrombopoiesis is spatially regulated by the bone marrow vasculature, Nat Commun. 2017 8(1):127. Figure. Figure. Disclosures No relevant conflicts of interest to declare.

Radial and longitudinal strain assessment in the carotid artery wall using speckle tracking

Pseudomonas aeruginosa is well adapted to the hospital setting and can cause a wide array of nosocomial infections that occasionally culminate in recalcitrant outbreaks. In the present study, we describe the first nosocomial outbreak of infection caused by blaVIM-2-positive P. aeruginosa in Germany. In November and December 2007, highly resistant P. aeruginosa isolates were recovered from the urine of 11 patients in the Department of Urology of a University Hospital. Bacterial isolates were typed by multilocus sequence typing and screened for known metallo-β-lactamase (MBL) genes by PCR. Environmental sources of transmission were tested for bacterial contamination using surveillance cultures. Furthermore, a matched case–control study was performed in search of medical procedures significantly associated with case status. Typing of recovered isolates confirmed VIM-2 MBL-producing P. aeruginosa of sequence type 175 in all cases. Surveillance cultures did not lead to the identification of an environmental source of the outbreak strain. Case–control analysis revealed retrograde urography as the only exposure significantly associated with case status. The analyses suggest the transmission of a single clone of VIM-2 MBL-producing P. aeruginosa leading to the infection of 11 patients within 47 days. Events in temporal proximity to retrograde urographies appear to have facilitated infection in the majority of cases. Department-specific infection control measures, including reinforced hygiene procedures during retrograde urography, quickly terminated the outbreak.

Fluorescence microscopy allows specific and selective imaging of biological samples. Unfortunately, unspecific background due to auto-fluorescence, scattering, and non-ideal labeling efficiency often adversely affect imaging. Surface plasmon-coupled emission (SPCE) is known to selectively mediate fluorescence that spatially originates from regions close to the metal interface. However, SPCE combined with fluorescence imaging has not been widely successful so far, most likely due to its limited photon yield, which makes it tedious to identify the exact window of the application. As the strength of SPCE based imaging is its unique sectioning capabilities. We decided to identify its clear beneficial operational regime for biological settings by interrogating samples in the presence of ascending background levels. For fluorescent beads as well as live-cell imaging as examples, we show how to extend the imaging performance in extremely high photon background environments. In a common setup using plasmonic gold-coated coverslips using an objective-based total internal reflection fluorescence microscope (TIRF-M), we theoretically and experimentally characterize our fluoplasmonics (f-Pics) approach by providing general user guidance in choosing f-Pics over TIRF-M or classical wide-field (WF).

Ethnopharmacological relevance The decoction of leaves of Clinopodium mexicanum Benth. Goaverts (Lamiaceae), commonly known as “Toronjil de Monte”, is used in the Mexican traditional medicine to induce sleep, as well as sedative and analgesic remedy. Aim of the study To evaluate the putative depressant effects of an aqueous extract of the medicinal plant Clinopodium mexicanum on the central nervous system (CNS). Materials and methods The effects of the extract (AECM) on mice were tested in several animal paradigms, including sodium pentobarbital-induced sleep, open field tests, and hole-board tests. The effects of AECM on pentylenetetrazole- and picrotoxin-induced convulsions in mice and on the antithermonociceptive response in the hot-plate paradigm were also tested. Additionally, the active extract (AECM) was analyzed with HPLC–ESI-MS techniques. Results Mice acutely treated with AECM at 100, 200, 500 and 1000 mg/kg doses prolonged the sleeping time induced by sodium pentobarbital (42 mg/kg). This extract, at 100 and 200 mg/kg doses, showed a sedative effect in the hole-board paradigm and decreased spontaneous activity in mice. AECM at 10, 100 and 200 mg/kg prolonged the onset of seizures induced by pentylenetetrazole (90 mg/kg) and antagonized tonic convulsions induced by picrotoxin (10 mg/kg). Additionally, AECM inhibited the response to a thermonociceptive stimulus. The intraperitoneal AECM treatment produced mortality with an LD50 = 2154 mg/kg. Chemical analysis showed that the flavanone glycosides neoponcirin, poncirin, and isonaringenin are the main compounds of the active extract. Conclusions This study demonstrates that an acutely administered single dose of an aqueous extract of Clinopodium mexicanum can exert depressant effects on the CNS. These findings are in agreement with the traditional use of Clinopodium mexicanum to induce sleep as well as sedative and analgesic remedy. The chemical analysis of AECM revealed the presence of the flavanone glycosides neoponcirin, poncirin, and isonaringin.

In this work we present the synthesis of branched gold nanoparticles based on the seed mediated growth technique. The materials have been characterized by advanced electron microscopy techniques in order to elucidate morphology, size and internal structure.

In den letzten Jahren ist der Lobbyismus wieder ins Gerede gekommen, manche Beobachter konstatieren eine Zasur hinsichtlich der institutionellen Trennung von Wirtschaft und Staat. Hat sich das Verhaltnis zwischen dem Staat und organisierten Interessen durch Expertenkommissionen und kommerzielle Beratung in der „Berliner Raterepublik“ nun grundlegend gewandelt und werden Berater zu zentralen Akteuren auf der politischen Buhne? Im ersten Schritt wird ein Uberblick uber die Organisationsformen und den Wandel des Lobbyismus, der immer weniger uber Verbande verlauft, gegeben und die unterschiedlichen Formen der Politikberatung vorgestellt. Die Grenzen zwischen legitimer Interessenvertretung durch Verbande und „verdeckte“ Schattenpolitik haben sich in den letzten Jahren gelockert und deshalb ist von einer Pluralisierung und neuen Unubersichtlichkeit der Politikberatung auszugehen. Vor dem Hintergrund eines Steuerungsverlustes der Politik werden die funktionalen Erfordernisse einer Einbeziehung nicht-staatlicher Akteure in das Politikmanagement erortert und auf die grundlegenden Probleme der Politikberatung, deren Reichweite und Grenzen eingegangen.

Between 1977 and 1984 a total of 301 patients with autonomous thyroid adenoma were irradiated with an individually calculated one-time dose of 400 Gy units of 131I. Repeated follow-up tests were made in 217 patients (up to 7.2 years, mean of two years). Pre- and post-treatment diagnosis in all patients consisted of determining T3 and T4, one TRH test, one 131I two-phase test to determine treatment, including quantified scintigraphy (under suppression, if necessary), as well as post-treatment scintigraphy and (post-treatment) 99mTc scintigraphy. The treatment was successful in 98% of patients; there was no difference between compensated and decompensated forms. Euthyroid state was achieved in 87% of patients, with typical findings of compensated T3 oversecretion, as is known to occur with endemic goiter in regions of iodine deficiency. The ability of the thyroid for autoregulatory adaptation to such iodine deficiency is thus preserved. Preclinical hypothyroidism occurred in 11% of patients: it could have been avoided in about half of them. Persistent or recurring autonomous adenoma was observed in 2% of patients as a result of under-dosage. One should thus aim at a dose of 400 Gy, to obtain optimal elimination. Radiation-induced carcinogenesis was not observed: radioiodine treatment and operation are thus of equal value in the causal treatment of autonomous adenoma. Radioiodine treatment is indicated in patients aged over 40 years with additional diseases and increased risk of anaesthesia and operation. It is preferred treatment if there are multiple autonomous adenomas.

Between 1980 and 1991, ionizing radiation was applied for analgesic purposes to 181 patients (97 men, 84 women, mean age 54 [29-81] years) with degenerative-inflammatory skeletal disease. The long-term effects were evaluated by questionnaire. Radiation of 2.5 to 6.0 Gy achieved lasting pain relief in 21 of 30 patients (70%) with arthritis of the shoulder or humeroscapular periarthritis, 15 of 21 (71%) with arthritis of the hip, in 12 of 15 (80%) with heel spurs or Achilles tendon bursitis and 10 of 11 (91%) with epicondylitis. Pain relief lasted for longer than two years in 41 of the 77 patients (53%). There were no side effects at the stated dosage. According to dose measurements the theoretical risk of malignant tumour induction is 20-40/million radiated patients and thus four orders of magnitude below the spontaneous malignant tumour incidence rate. The genetic risk is even lower. Ionizing radiation of degenerative-inflammatory diseases is thus an effective form of treatment with few side effects.

Bislang lauft der Diskurs zur Digitalisierung in Deutschland nach dem traditionellen Muster: Erst werden die Herausforderungen verdrangt, und wenn es fast zu spat fur eine rationale Steuerung des Prozesses ist, setzt Panik ein. Diese Phase ist inzwischen erreicht, und die Debatte schwankt zwischen euphorischen Hoffnungen etwa in die „Industrie 4.0“, die unseren Produktionsstandort vielleicht doch noch retten konnte, bis hin zu den soziologischen KritikerInnen, die im Rahmen ihrer Warnungen vor einem unkontrollierten Finanzkapitalismus und der Okonomisierung der Lebenswelten die Risiken von „Big Data“ oft mit deutlichen Worten markieren.

Fluorescence correlation spectroscopy (FCS) is an ideal analytical tool for studying concentrations, propagation, interactions and internal dynamics of molecules at nanomolar concentrations in living cells. FCS analyzes minute fluorescence-intensity fluctuations about the equilibrium of a small ensemble (10(3)) of molecules. These fluctuations act like a 'fingerprint' of a molecular species detected when entering and leaving a femtoliter-sized optically defined observation volume created by a focused laser beam. In FCS the fluorescence fluctuations are recorded as a function of time and then statistically analyzed by autocorrelation analysis. The resulting autocorrelation curve yields a measure of self-similarity of the system after a certain time delay, and its amplitude describes the normalized variance of the fluorescence fluctuations. By fitting the curves to an appropriate physical model, this method provides precise information about a multitude of measurement parameters, including diffusion coefficients, local concentration, states of aggregation and molecular interactions. FCS operates in real time with diffraction-limited spatial and sub-microsecond temporal resolution. Assessing diverse molecular dynamics within the living cell is a challenge well met by FCS because of its single-molecule sensitivity and high dynamic resolution. For these same reasons, however, intracellular FCS measurements also harbor the large risk of collecting artifacts and thus producing erroneous data. Here we provide a step-by-step guide to the application of FCS to cellular systems, including methods for minimizing artifacts, optimizing measurement conditions and obtaining parameter values in the face of diverse and complex conditions of the living cell. A discussion of advantages and disadvantages of one-photon versus two-photon excitation for FCS is available in Supplementary Methods online.

Assessing the dynamics of individual membrane proteins in living cells is a powerful approach to investigate their assembly, mobility, and function. Here, we describe how to image single G protein-coupled receptors (GPCRs), both in the active and inactive state. This is achieved by combining labeling of GPCRs with bright organic fluorophores and fluorescent imaging by total internal reflection fluorescence microscopy. Using this method, individual tracks of single molecules can be analyzed in parallel with high spatial precision and with frame rates up to 50/s.

Wiskott-Aldrich syndrome (WAS) is caused by mutations in the WAS gene and is characterized by immunodeficiency, eczema and microthrombocytopenia. The molecular link between WAS mutations and microthrombocytopenia is unknown. Profilin1 (Pfn1) is a key actin-regulating protein that, besides actin, interacts with phosphoinositides and multiple proline-rich proteins, including the WAS protein (WASp)/WASp-interacting protein (WIP) complex. Here we report that mice with a megakaryocyte/platelet-specific Pfn1 deficiency display microthrombocytopenia due to accelerated turnover of platelets and premature platelet release into the bone marrow. Both Pfn1-null mouse platelets and platelets isolated from WAS patients contained abnormally organized and hyperstable microtubules. These results reveal an unexpected function of Pfn1 as a regulator of microtubule organization and point to a previously unrecognized mechanism underlying the platelet formation defect in WAS patients.