65 research outputs found
Quantitative performance characterization of three-dimensional noncontact fluorescence molecular tomography
© 2016 The Authors.Fluorescent proteins and dyes are routine tools for biological research to describe the behavior of genes, proteins, and cells, as well as more complex physiological dynamics such as vessel permeability and pharmacokinetics. The use of these probes in whole body in vivo imaging would allow extending the range and scope of current biomedical applications and would be of great interest. In order to comply with a wide variety of application demands, in vivo imaging platform requirements span from wide spectral coverage to precise quantification capabilities. Fluorescence molecular tomography (FMT) detects and reconstructs in three dimensions the distribution of a fluorophore in vivo. Noncontact FMT allows fast scanning of an excitation source and noninvasive measurement of emitted fluorescent light using a virtual array detector operating in free space. Here, a rigorous process is defined that fully characterizes the performance of a custom-built horizontal noncontact FMT setup. Dynamic range, sensitivity, and quantitative accuracy across the visible spectrum were evaluated using fluorophores with emissions between 520 and 660 nm. These results demonstrate that high-performance quantitative three-dimensional visible light FMT allowed the detection of challenging mesenteric lymph nodes in vivo and the comparison of spectrally distinct fluorescent reporters in cell culture
A Limited Role for the Cell Cycle Regulator Cyclin A1 in Murine Leukemogenesis
The quest for novel therapeutic targets in acute myeloid leukemia (AML) is still ongoing. One of such targets, cyclin A1, was shown to be overexpressed in AML including AML stem cells. However, the function of cyclin A1 in AML is largely unknown, and the data on its impact on patientsŽ survival remain controversial. Therefore, we developed a transgenic mouse model of stem cell-directed inducible cyclin A1 overexpression and crossed these mice with PML-RARα-knockin mice, which develop an AML M3-like phenotype. To observe the effects of cyclin A1 loss-of-function, we also crossed PML-RARα-knockin mice to cyclin A1-knockout mice. Neither overexpression nor loss of cyclin A1 significantly altered leukemogenesis in PML-RARα-knockin mice. These findings imply that upregulation of cyclin A1 is not essential for leukemogenesis. Our data suggest that cyclin A1 does not represent a suitable target for AML therapy
Detrital garnet petrology challenges Paleoproterozoic ultrahigh-pressure metamorphism in western Greenland
Modern-style plate tectonics is characterised by the
global operation of cold and deep subduction involving blueschist facies and
ultrahigh-pressure metamorphism. This has been a common process since the
Neoproterozoic, but a couple of studies indicate similar processes were
active in the Paleoproterozoic, at least on the local scale. Particularly
conspicuous are extreme ultrahigh-pressure conditions of âŒâ7âGPa at thermal gradients <â150ââCâGPaâ1 proposed for
metamorphic rocks of the Nordre StrĂžmfjord shear zone in the western part
of the Paleoproterozoic Nagssugtoqidian Orogen of Greenland. By acquiring a
large dataset of heavy minerals (nâ=â52â130) and garnet major-element
composition integrated with mineral inclusion analysis (n=2669) from
modern sands representing fresh and naturally mixed erosional material from
the metamorphic rocks, we here intensely screened the area for potential
occurrences of ultrahigh-pressure rocks and put constraints on the
metamorphic evolution. Apart from the absence of any indications pointing to
ultrahigh-pressure and low-temperatureâhigh-pressure metamorphism, the
results are well in accordance with a common Paleoproterozoic
subductionâcollision metamorphic evolution along a Barrovian-type
intermediate temperature and pressure gradient with a pressure peak at the
amphiboliteâgranuliteâeclogite-facies transition and a temperature peak
at medium- to high-pressure granulite-facies conditions. In addition, we
discuss that all âevidenceâ for ultrahigh-pressure metamorphism proposed
in the literature for rocks of this area is equivocal. Accordingly, the
Nordre StrĂžmfjord shear zone is not an example of modern-style plate
tectonics in the Paleoproterozoic or of very low thermal gradients and
extreme pressure conditions in general.</p
Clinical Processes - The Killer Application for Constraint-Based Process Interactions?
For more than a decade, the interest in aligning information
systems in a process-oriented way has been increasing. To enable operational
support for business processes, the latter are usually specified in
an imperative way. The resulting process models, however, tend to be too
rigid to meet the flexibility demands of the actors involved. Declarative
process modeling languages, in turn, provide a promising alternative in
scenarios in which a high level of flexibility is demanded. In the scientific
literature, declarative languages have been used for modeling rather simple
processes or synthetic examples. However, to the best of our knowledge,
they have not been used to model complex, real-world scenarios
that comprise constraints going beyond control-flow. In this paper, we
propose the use of a declarative language for modeling a sophisticated
healthcare process scenario from the real world. The scenario is subject to
complex temporal constraints and entails the need for coordinating the
constraint-based interactions among the processes related to a patient
treatment process. As demonstrated in this work, the selected real process
scenario can be suitably modeled through a declarative approach.Ministerio de EconomĂa y Competitividad TIN2016-76956-C3-2-RMinisterio de EconomĂa y Competitividad TIN2015-71938-RED
Tetracycline Inducible Gene Manipulation in Serotonergic Neurons
The serotonergic (5-HT) neuronal system has important and diverse physiological functions throughout development and adulthood. Its dysregulation during development or later in adulthood has been implicated in many neuropsychiatric disorders. Transgenic animal models designed to study the contribution of serotonergic susceptibility genes to a pathological phenotype should ideally allow to study candidate gene overexpression or gene knockout selectively in serotonergic neurons at any desired time during life. For this purpose, conditional expression systems such as the tet-system are preferable. Here, we generated a transactivator (tTA) mouse line (TPH2-tTA) that allows temporal and spatial control of tetracycline (Ptet) controlled transgene expression as well as gene deletion in 5-HT neurons. The tTA cDNA was inserted into a 196 kb PAC containing a genomic mouse Tph2 fragment (177 kb) by homologous recombination in E. coli. For functional analysis of Ptet-controlled transgene expression, TPH2-tTA mice were crossed to a Ptet-regulated lacZ reporter line (Ptet-nLacZ). In adult double-transgenic TPH2-tTA/Ptet-nLacZ mice, TPH2-tTA founder line L62-20 showed strong serotonergic ÎČ-galactosidase expression which could be completely suppressed with doxycycline (Dox). Furthermore, Ptet-regulated gene expression could be reversibly activated or inactivated when Dox was either withdrawn or added to the system. For functional analysis of Ptet-controlled, Cre-mediated gene deletion, TPH2-tTA mice (L62-20) were crossed to double transgenic Ptet-Cre/R26R reporter mice to generate TPH2-tTA/Ptet-Cre/R26R mice. Without Dox, 5-HT specific recombination started at E12.5. With permanent Dox administration, Ptet-controlled Cre-mediated recombination was absent. Dox withdrawal either postnatally or during adulthood induced efficient recombination in serotonergic neurons of all raphe nuclei, respectively. In the enteric nervous system, recombination could not be detected. We generated a transgenic mouse tTA line (TPH2-tTA) which allows both inducible and reversible transgene expression and inducible Cre-mediated gene deletion selectively in 5-HT neurons throughout life. This will allow precise delineation of serotonergic gene functions during development and adulthood
Designing a Process Mining-Enabled Decision Support System for Business Process Standardization in ERP Implementation Projects
Process standardization allows to optimize ERP systems and is a nec-essary step prior to ERP implementation projects. Traditional approaches to standardizing business processes are based on manually created "de-jure" process models, which are distorted, error-prone, simplistic, and often deviating from process reality. Theoretically embedded in the organizational contingency theory as kernel theory, this paper employs a design science approach to design a process mining-enabled decision support system (DSS) which combines bottom-up process mining models with manually added top-down standardization infor-mation to recommend a suitable standard process specification from a repository. Extended process models of the as-is process are matched against a repository of best-practice standard process model using an attributebased process similarity matching algorithm. Thus, the DSS aims to reduce the overall costs of process standardization, to optimize the degree of fit between the organization and the implemented processes, and to minimize the degree of organizational change re-quired in standardization and ERP implementation projects. This paper imple-ments a working prototype instantiation in the open-source process analytics platform Apromore based on a real-life event log and standardization attributes for the Purchase-to-Pay and Order-to-Cash processes from three SAP R/3 ERP systems at the industry partner
Inducible Gene Manipulations in Brain Serotonergic Neurons of Transgenic Rats
The serotonergic (5-HT) system has been implicated in various physiological processes and neuropsychiatric disorders, but in many aspects its role in normal and pathologic brain function is still unclear. One reason for this might be the lack of appropriate animal models which can address the complexity of physiological and pathophysiological 5-HT functioning. In this respect, rats offer many advantages over mice as they have been the animal of choice for sophisticated neurophysiological and behavioral studies. However, only recently technologies for the targeted and tissue specific modification of rat genes - a prerequisite for a detailed study of the 5-HT system - have been successfully developed. Here, we describe a rat transgenic system for inducible gene manipulations in 5-HT neurons. We generated a Cre driver line consisting of a tamoxifen-inducible CreERT2 recombinase under the control of mouse Tph2 regulatory sequences. Tissue-specific serotonergic Cre recombinase expression was detected in four transgenic TPH2-CreERT2 rat founder lines. For functional analysis of Cre-mediated recombination, we used a rat Cre reporter line (CAG-loxP.EGFP), in which EGFP is expressed after Cre-mediated removal of a loxP-flanked lacZ STOP cassette. We show an in-depth characterisation of this rat Cre reporter line and demonstrate its applicability for monitoring Cre-mediated recombination in all major neuronal subpopulations of the rat brain. Upon tamoxifen induction, double transgenic TPH2-CreERT2/CAG-loxP.EGFP rats show selective and efficient EGFP expression in 5-HT neurons. Without tamoxifen administration, EGFP is only expressed in few 5-HT neurons which confirms minimal background recombination. This 5-HT neuron specific CreERT2 line allows Cre-mediated, inducible gene deletion or gene overexpression in transgenic rats which provides new opportunities to decipher the complex functions of the mammalian serotonergic system
Afrikaans as Standaard Gemiddelde Europees:Wanneer ân lid uit sy taalarea beweeg
A recent trend in the study of Standard Average European is the extraterritorial perspective of examining the extent to which non-European languages have converged with this Sprachbund as a result of contact with one or more of its members. The present article complements this line of research in that it investigates the extent to which a European language has diverged from Standard Average European after leaving the linguistic area. The focus is on Dutch, a nuclear member of the Sprachbund, and Afrikaans, its colonial offshoot. The two languages are compared with respect to twelve of the most distinctive linguistic features of Standard Average European. Afrikaans is found to share ten of them with Dutch, including anticausative prominence and formally distinguished intensifiers and reflexives, and could therefore still be considered a core member of the Sprachbund, despite deviations in the expression of negative pronouns and the grammaticality of external possessor constructions. This relatively low degree of divergence may be attributed to the continuity from Settler Dutch to at least the variety of Afrikaans on which the standard language is based and to the important role that Dutch continued to play in the history of Afrikaans
New Mouse Lines for the Analysis of Neuronal Morphology Using CreER(T)/loxP-Directed Sparse Labeling
BACKGROUND: Pharmacologic control of Cre-mediated recombination using tamoxifen-dependent activation of a Cre-estrogen receptor ligand binding domain fusion protein [CreER(T)] is widely used to modify and/or visualize cells in the mouse. METHODS AND FINDINGS: We describe here two new mouse lines, constructed by gene targeting to the Rosa26 locus to facilitate Cre-mediated cell modification. These lines should prove particularly useful in the context of sparse labeling experiments. The R26rtTACreER line provides ubiquitous expression of CreER under transcriptional control by the tetracycline reverse transactivator (rtTA); dual control by doxycycline and tamoxifen provides an extended dynamic range of Cre-mediated recombination activity. The R26IAP line provides high efficiency Cre-mediated activation of human placental alkaline phosphatase (hPLAP), complementing the widely used, but low efficiency, Z/AP line. By crossing with mouse lines that direct cell-type specific CreER expression, the R26IAP line has been used to produce atlases of labeled cholinergic and catecholaminergic neurons in the mouse brain. The R26IAP line has also been used to visualize the full morphologies of retinal dopaminergic amacrine cells, among the largest neurons in the mammalian retina. CONCLUSIONS: The two new mouse lines described here expand the repertoire of genetically engineered mice available for controlled in vivo recombination and cell labeling using the Cre-lox system
Assessment of ecosystem services of an urbanized tropical estuary with a focus on habitats and scenarios
Tropical estuaries are one of the most valuable ecosystems on the planet because of the number of ecosystem services they provide. The increasing anthropogenic pressure to which these estuaries are subject has caused a reduction in their natural capital stock. Therefore, the application of a pragmatic and rational ecosystem-based management approach to sustainably manage the multiple ecosystem services provided by this ecosystem is necessary. The aim of our study is to present an approach that combines prospective scenarios with habitat-based perspective to assess the supply capacity of ecosystem services, plus determine the impact of protected areas in an urbanized tropical estuary. The current situation and two scenarios were generated to evaluate the capacity of habitats to supply ecosystem services. This type of assessment will allow the decision makers to visualize the effect of their choices or the occurrence of events which might produce significant changes in the estuary. Thus, over time, measures can be taken to sustain the supply of ecosystem services. We determined that the establishment of protected areas have a positive impact; however, the effect is not the same for all of them. Consequently, indicating that actions such as community participation, research, education, management planning and infrastructure development must accompany the development of a protected area
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