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Abstract The transcription factor regulatory factor X 7 (RFX7) has been identified as a tumor suppressor that is recurrently mutated in lymphoid cancers and appears to be dysregulated in many other cancers. RFX7 is activated by the well-known tumor suppressor p53 and regulates several other known tumor suppressor genes. However, what other factors regulate RFX7 and its target genes remains unclear. Here, reporter gene assays were used to identify that RFX7 regulates the tumor suppressor gene PDCD4 through direct interaction with its X-box promoter motif. We utilized mass spectrometry to identify factors that bind to DNA together with RFX7. In addition to RFX7, we also identified RFX5, RFXAP, RFXANK, and ANKRA2 that bind to the X-box motif in the PDCD4 promoter. We demonstrate that ANKRA2 is a bona fide direct p53 target gene. We used transcriptome analyses in two cell systems to identify genes regulated by ANKRA2, its sibling RFXANK, and RFX7. These results revealed that ANKRA2 functions as a critical cofactor of RFX7, whereas RFXANK regulates largely distinct gene sets.

This paper presents a first conceptualization for decentralized project delivery through the combination of blockchain technology and common pool resource governance theory, also known as the ”crypto commons”. While previous literature on project delivery models identifies the use of decentralized governance, there is not yet a holistic conceptualization nor a specific overview of governance applications that can be applied. To develop the concept, we use a literature review to synthesize fourteen blockchain governance mechanisms useful for managing the crypto-commons. Subsequently, we use systematic combining to identify twenty-two specific applications for decentralized project delivery in the construction industry, one of the largest project-based industries. Each application is briefly reviewed, and examples of their relevance to realize decentralized project delivery models are provided. We discuss the potential for novel forms of project delivery, but also the need for future research on the applications, as well as on the system level implications, the system design challenges, and the implementation barriers in the specific context of the construction industry. Overall, the concepts and mechanisms presented provide a theoretical foundation upon which future researchers can use to design novel decentralized project delivery models.

Abstract Rapid renovation of Europe’s inefficient buildings is required to reduce climate change. However, evaluating buildings at scale is challenging because every building is unique. In current practice, the energy performance of buildings is assessed during on-site visits, which are slow, costly, and local. This paper presents a building point cloud dataset that promotes a data-driven, large-scale understanding of the 3D representation of buildings and their energy characteristics. We generate building point clouds by intersecting building footprints with geo-referenced LiDAR data and link them with attributes from UK’s energy performance database via the Unique Property Reference Number (UPRN). To mimic England’s building stock’s features well, we select one million buildings from a range of rural and urban regions, of which half a million are linked to energy characteristics. Building point clouds in new regions can be generated with our published open-source code. The dataset enables novel research in building energy modeling and can be easily expanded to other research fields by adding building features via the UPRN or geo-location.

The role of long non‐coding RNAs (lncRNAs) in p53‐mediated tumor suppression has become increasingly appreciated in the past decade. Thus, the identification of p53‐regulated lncRNAs can be a promising starting point to select and prioritize lncRNAs for functional analyses. By integrating transcriptome and transcription factor‐binding data, we identified 379 lncRNAs that are recurrently differentially regulated by p53. Dissecting the mechanisms by which p53 regulates many of them, we identified sets of lncRNAs regulated either directly by p53 or indirectly through the p53‐RFX7 and p53‐p21‐DREAM/RB:E2F pathways. Importantly, we identified multiple p53‐responsive lncRNAs that are co‐regulated with their protein‐coding host genes, revealing an important mechanism by which p53 may regulate lncRNAs. Further analysis of transcriptome data and clinical data from cancer patients showed that recurrently p53‐regulated lncRNAs are associated with patient survival. Together, the integrative analysis of the landscape of p53‐regulated lncRNAs provides a powerful resource facilitating the identification of lncRNA function and displays the mechanisms of p53‐dependent regulation that could be exploited for developing anticancer approaches.

Abstract Recurrently mutated in lymphoid neoplasms, the transcription factor RFX7 is emerging as a tumor suppressor. Previous reports suggested that RFX7 may also have a role in neurological and metabolic disorders. We recently reported that RFX7 responds to p53 signaling and cellular stress. Furthermore, we found RFX7 target genes to be dysregulated in numerous cancer types also beyond the hematological system. However, our understanding of RFX7’s target gene network and its role in health and disease remains limited. Here, we generated RFX7 knock-out cells and employed a multi-omics approach integrating transcriptome, cistrome, and proteome data to obtain a more comprehensive picture of RFX7 targets. We identify novel target genes linked to RFX7’s tumor suppressor function and underscoring its potential role in neurological disorders. Importantly, our data reveal RFX7 as a mechanistic link that enables the activation of these genes in response to p53 signaling.

Driving requires continuous decision making from a driver taking into account all available relevant information. Automating driving tasks also automates the related decisions. However, humans are very good at dealing with bad quality, fuzzy, informal and incomplete information, whereas machines generally require solid quality information in a formalized format. Therefore, the development of automated driving functions relies on the availability of machine-usable information. A digital twin contains quality controlled information collected and augmented from different sources, ready to be supplied to such an automated driving function. An information model that describes all conceivably relevant information is necessary. To this end, a list of requirements that such an information model should meet is proposed and each requirement is argued for. Based on the anticipated services and applications that such a system should support, a collection of requirements for system architecture is derived. Information modeling is performed for selected relevant information groups. A system architecture has been proposed and validated with three different implementations, addressing several different applications to support decisions at a highway tunnel construction site in Austria and throughout the Test Bed Lower Saxony in Germany.

Abstract Cellular senescence is a stable cell cycle arrest that normal cells undergo after a finite number of divisions, in response to a variety of intrinsic and extrinsic stimuli. Although senescence is largely established and maintained by the p53/p21WAF1/CIP1 and pRB/p16INK4A tumour suppressor pathways, the downstream targets responsible for the stability of the growth arrest are not known. We have employed a stable senescence bypass assay in conditionally immortalised human breast fibroblasts (CL3EcoR) to investigate the role of the DREAM complex and its associated components in senescence. DREAM is a multi-subunit complex comprised of the MuvB core, containing LIN9, LIN37, LIN52, LIN54, and RBBP4, that when bound to p130, an RB1 like protein, and E2F4 inhibits cell cycle-dependent gene expression thereby arresting cell division. Phosphorylation of LIN52 at Serine 28 is required for DREAM assembly. Re-entry into the cell cycle upon phosphorylation of p130 leads to disruption of the DREAM complex and the MuvB core, associating initially to B-MYB and later to FOXM1 to form MMB and MMB-FOXM1 complexes respectively. Here we report that simultaneous expression of MMB-FOXM1 complex components efficiently bypasses senescence with LIN52, B-MYB, and FOXM1 as the crucial components. Moreover, bypass of senescence requires non-phosphorylated LIN52 that disrupts the DREAM complex, thereby indicating a central role for assembly of the DREAM complex in senescence.

Although the prognosis in Guillain-Barré syndrome (GBS) is generally good, protracted and incomplete courses of recovery can be a heavy burden. Animal studies suggest growth hormone (GH) treatment could stimulate myelin repair and thus accelerate functional recovery in acute polyneuropathy. We report on the first use of GH in GBS. Our objective was to monitor safety and tolerability as well as to evaluate the effect of an off-label GH therapy during recovery from GBS in 1 patient. A 28-year-old male with flaccid tetraparesis caused by pure motor GBS was treated off-label with GH (1 mg/day) for 10 weeks. Muscle strength was measured regularly before, during, and after the treatment over a total span of 330 days. Serum levels of IGF-I were assessed before, during, and after GH treatment. Changes in strength gain were used as the main parameter of efficacy. No side effects of GH treatment were observed. Serum IGF-I increased from 177 ng/mL at baseline to an average of 342 ng/mL (normal range 78–270 ng/mL) during treatment. Prior to GH administration, strength (R2 = 0.99, p < 0.01) was associated with time, representing the natural course of recovery. During GH treatment, the slope of strength gain increased (Glass’ ∆ = 1.08, p < 0.01). The association between alterations of strength gain and IGF-I serum levels reached trend level (R2 = 0.36, p = 0.09). In this single case, GH treatment seemed to be associated with faster muscular strength gain. Controlled studies are needed in order to establish GH as a potential therapeutic approach in motor GBS.

Objectives: In the microbiological diagnosis of periprosthetic joint infection (PJI), there is much discussion about the methodology of obtaining proper specimens, the processing technique, and suitable culture media. This retrospective study was conducted to analyse the accuracy of our culture techniques. Methods: Tissue samples and components from 258 patients after revision arthroplasty of the hip, knee, and shoulder were investigated, and the results of tissue cultures (TC) were compared to those of sonicate fluid cultures (SFC). Furthermore, an evaluation was performed of the influence of different culture media on the detection rate. Results: PJI was confirmed in 186 patients. The overall sensitivity of TC was no different to that of SFC (91.3% vs 90.8%, P = 1). In 153 cases (82.3%), TC and SFC showed concordant positive results. Results were discordant in 33 cases (17.7%). When differentiated according to the type of infection, TC showed significantly better results than SFC in detecting polymicrobial infections (97.0% vs 67.0%, P = 0.004). There were also significant differences between the culture media regarding the yield of microorganisms. Conclusions: TC was more effective in detecting co-infections. The best results were obtained using both TC and SFC. The choice of culture media has a significant influence on the quality of results.

Septins are part of the cytoskeleton and polymerize into non-polar filaments of heteromeric hexamers or octamers. They belong to the class of P-loop GTPases but the roles of GTP binding and hydrolysis on filament formation and dynamics are not well understood. The basic human septin building block is the septin rod, a hetero-octamer composed of SEPT2, SEPT6, SEPT7, and SEPT9 with a stoichiometry of 2:2:2:2 (2-6-7-9-9-7-6-2). Septin rods polymerize by end-to-end and lateral joining into linear filaments and higher ordered structures such as rings, sheets, and gauzes. We purified a recombinant human septin octamer from E. coli for in vitro experimentation that is able to polymerize into filaments. We could show that the C-terminal region of the central SEPT9 subunit contributes to filament formation and that the human septin rod decreases the rate of in vitro actin polymerization. We provide further first kinetic data on the nucleotide uptake- and exchange properties of human hexameric and octameric septin rods. We could show that nucleotide uptake prior to hydrolysis is a dynamic process and that a bound nucleotide is exchangeable. However, the hydrolyzed γ-phosphate is not released from the native protein complex. We consequently propose that GTP hydrolysis in human septins does not follow the typical mechanism known from other small GTPases.

miR-132 was shown to drive pathological cardiac remodeling, a hallmark of heart failure. Here, the authors show that an antisense inhibitor of miR-132 has favourable pharmacokinetics, safety-tolerability and preclinical efficacy in mouse and porcine models of heart failure.

Often, research on the efficacy of postsecondary workforce programs does not convey their impact on true social mobility. The purpose of this study is to investigate project-based Career and Technical Education (CTE) workforce pathways in Silicon Valley. This study takes a step towards better understanding what constitutes the metrics that explain functioning pathways. In contributing to Project-Based Learning (PBL) theory, Amaral et al. (2015) found that seven PBL essentials form good learning outcomes; Creghan and Adair-Creghan (2015) then showed a measurable outcome of PBL is higher attendance, to which Plasman and Gottfried (2020), using a case of Applied STEM CTE (AS-CTE), framed attendance as a predictor of the efficacy of a workforce pathway. Recommendation: Through ethnography, the investigators observed that when social mobility was added as a metric of high quality PBL with AS-CTE in a predictive ontology framework of education success, an improved level of attendance was observed. The authors conclude that using the seven essentials and social mobility as a metric of PBL helps explain the observation of PBL’s improved efficacy. Hence, social mobility should be a metric of PBL AS-CTE program outcomes.

Compared with other industries, construction has maintained one of the lowest growth rates in productivity and innovation over the last decades. Moreover, low compliance with schedule targets has remained a common issue throughout projects. Therefore, although methodologies developed have benefited productivity in design, construction and operation, meeting client and project objectives becomes challenging without properly integrating organization, information, systems and processes. This perspective introduces the Virtual Design and Construction (VDC) framework. First, this paper studies and reviews VDC since its creation at the Center for Integrated Facility Engineering (CIFE) of Stanford University in 2001. After that, a comprehensive bibliographical review of the VDC framework is presented and updated with examples of its application in construction projects, delving into its elements: Client Objectives, Project Objectives, Production Objectives, Controllable Factors, Integrated Concurrent Engineering (ICE), Building Information Modeling (BIM) and Project Production Management (PPM). Finally, the main benefits and project outcomes when applying the VDC framework are presented and discussed, followed by the conclusions and recommendations for future research.

The tumor suppressor p53 and its oncogenic sibling p63 (ΔNp63) direct opposing fates in tumor development. These paralog proteins are transcription factors that elicit their tumor suppressive and oncogenic capacity through the regulation of both shared and unique target genes. Both proteins predominantly function as activators of transcription, leading to a paradigm shift away from ΔNp63 as a dominant negative to p53 activity. The discovery of p53 and p63 as pioneer transcription factors regulating chromatin structure revealed new insights into how these paralogs can both positively and negatively influence each other to direct cell fate. The previous view of a strict rivalry between the siblings needs to be revisited, as p53 and p63 can also work together toward a common goal.

Summary: Aiming at clinical translation, robust directed differentiation of human pluripotent stem cells (hPSCs), preferentially in chemically defined conditions, is a key requirement. Here, feasibility of suspension culture based hPSC-cardiomyocyte (hPSC-CM) production in low-cost, xeno-free media compatible with good manufacturing practice standards is shown. Applying stirred tank bioreactor systems at increasing dimensions, our advanced protocol enables routine production of about 1 million hPSC-CMs/mL, yielding ∼1.3 × 108 CM in 150 mL and ∼4.0 × 108 CMs in 350–500 mL process scale at >90% lineage purity. Process robustness and efficiency is ensured by uninterrupted chemical WNT pathway control at early stages of differentiation and results in the formation of almost exclusively ventricular-like CMs. Modulated WNT pathway regulation also revealed the previously unappreciated role of ROR1/CD13 as superior surrogate markers for predicting cardiac differentiation efficiency as soon as 72 h of differentiation. This monitoring strategy facilitates process upscaling and controlled mass production of hPSC derivatives. : Directed differentiation of human pluripotent stem cell cardiomyocytes (CMs) is required for clinical translation. Here, xeno-free suspension culture production in stirred bioreactors is shown to enable routine generation of ∼1.3 × 108 CMs/150 mL and ∼4.0 × 108 CMs/500 mL scale at >90% purity. Uninterrupted chemical WNT pathway control and the previously unappreciated role of ROR1/CD13 as predictive differentiation markers facilitate large-scale production of ventricular-like CMs. Keywords: human pluripotent stem cells, chemically defined process, cardiac differentiation, cardiomyocyte, suspension culture, stirred tank bioreactor, ROR1, WNT, process development, upscaling

Abstract Background Non-specific chronic low back pain (nscLBP) has a high socio-economic relevance due to its high incidence, prevalence and associated costs. Therefore, it is essential to evaluate effective therapeutic strategies. This study examines the effects of moderate mountain exercise and spa therapy on orthopedic and psychophysiological parameters. Based on a three-armed randomized controlled trial, guided mountain hiking tours and balneotherapy in thermal water were compared to a control group. Methods Eighty patients with diagnosed nscLBP were separated into three groups: The two intervention groups GE (green exercise) and GEBT (green exercise and balneotherapy) undertook daily mountain hiking tours, whereas the GEBT group got an additional treatment with baths in Mg-Ca-SO4 thermal water. The third group (CO) received no intervention. GE and GEBT group were treated for 6 days; all groups were followed up for 120 days. Results Compared to GE and CO group, the GEBT treatment showed significant improvements of pain, some orthopedic parameters, health-related quality of life and mental well-being in patients with nscLBP. Conclusions The results of this study confirmed a benefit of mountain hiking combined with Mg-Ca-SO4 spa therapy as a multimodal treatment of patients with nscLBP. Further studies should focus on long-term-effects of this therapeutic approach. Trial registration ISRCTN, ISRCTN99926592. Registered 06. July 2018 - Retrospectively registered.

Abstract AiKomPass is a web-based tool to make informal and non-formal competences within the electrical and metal industries visible. It is based on an interview study with potential users and a task inventory, which covers tasks of work areas like process planning, production, maintenance and logistics within production processes. The tool itself was developed within a participative design process. It enables employees and people in search of employment to self-assess their ability to fulfil working tasks and thus visualizes their skills and knowledge. In addition it helps to analyse biographical aspects of competences (competences from spheres outside work and outside the electrical and metal industries and related certificates). The online tool focuses especially on people who are not formally qualified for the labour market. AiKomPass is available online (www.aikompass.de) and can be used free of charge.

Medical faculties have the responsibility to train tomorrow's doctors and in a crisis face the challenge of delivering students into the workforce promptly and safely. Worldwide, medical faculties have faced unprecedented disruptions from viral outbreaks and pandemics including SARS, Ebola, H1N1 and COVID-19 which bring unique challenges. Currently there is worldwide disruption to medical faculties and medical education due to COVID-19. Despite close links with clinical medicine and the known risks of pandemics, many medical faculties have been caught off guard without pandemic planning in place, to deal with an exponential rise in infections and deaths, overwhelmed health services and widespread community risk of transmission. Assessing transmission risk of COVID-19 in teaching, clinical and community attachments and continuing medical education is paramount as medical faculties face subsequent pandemics waves. Consensus statements based on best available evidence and international expertise from medical faculties in Asia, Australia and Europe were developed to help guide the protection of staff and students, priorities on teaching activities and further educational development. Infection prevention, infection control, contact tracing and medical surveillance are detailed to minimise transmission and to enhance safety. Recommendations on teaching activities planning can enhance responsiveness of medical faculties to tackle subsequent waves of COVID-19 infection. A global approach and dialogue are encouraged.

The transcription factor p53 is the best-known tumor suppressor, but its sibling p63 is a master regulator of epidermis development and a key oncogenic driver in squamous cell carcinomas (SCC). Despite multiple gene expression studies becoming available, the limited overlap of reported p63-dependent genes has made it difficult to decipher the p63 gene regulatory network. Particularly, analyses of p63 response elements differed substantially among the studies. To address this intricate data situation, we provide an integrated resource that enables assessing the p63-dependent regulation of any human gene of interest. We use a novel iterative de novo motif search approach in conjunction with extensive ChIP-seq data to achieve a precise global distinction between p53-and p63-binding sites, recognition motifs, and potential co-factors. We integrate these data with enhancer:gene associations to predict p63 target genes and identify those that are commonly de-regulated in SCC representing candidates for prognosis and therapeutic interventions.

Judging by the small number of published cases, periprosthetic joint infections (PJI) caused by Mycoplasma species are regarded as unusual. This is not surprising as special growth conditions are necessary for diagnosis and therefore the laboratory must be informed of any clinical suspicion. However, surgeons are generally not aware of the risk factors associated with certain microorganisms causing an infection. Our laboratory therefore decided to adopt a new strategy: first, to address specific questions concerning the medical history of the patient and second, to make diagnosis of rare and fastidious microorganisms part of routine investigation, even if detailed information is not available. Keywords: Mycoplasma hominis, Periprosthetic joint infection, Microbiological diagnosis, Antimicrobial susceptibility

Aim of the study was to investigate dynamic radio-ulnar incongruence (dRUI) in the canine elbow joint comparing orthopedic healthy and dysplastic dogs in a prospective in vivo study design. In 6 orthopedic sound elbow joints (5 dogs, median age 17 months & mean body weight 27.9 kg) and 7 elbow joints with medial coronoid disease (6 dogs, median age 17.5 months & mean body weight 27.6 kg) 0.8 mm Ø tantalum beads were surgically implanted into radius, ulna and humerus for dynamic radiosteriometric analysis (RSA) using high-speed biplanar fluoroscopy with the dogs walking on a treadmill. dRUI, in the form of proximo-distal translation of the radius relative to the ulna, was measured for the first third of stance phase and compared between groups using unpaired t-testing. Healthy elbow joints exhibited a relative radio-ulnar translation of 0.7 mm (SD 0.31 mm), while dysplastic joints showed a translation of 0.5 mm (SD 0.30 mm). No significant difference between groups was detected (p = 0.2092, confidence interval -0.6 – 0.2). Based on these findings dRUI is present in every canine elbow joint, as part of the physiological kinematic pattern. However, dysplastic elbow joints do not show an increased radio-ulnar translation, and therfore dRUI cannot be considered causative for medial coronoid disease.

Abstract High risk human papilloma viruses cause several types of cancer. The HPV oncoproteins E6 and E7 are essential for oncogenic cell transformation. E6 mediates the degradation of the tumor suppressor p53, and E7 can form complexes with the retinoblastoma pRB tumor suppressor. Recently, it has been shown that HPV E7 can also interfere with the function of the DREAM transcriptional repressor complex. Disruption of DREAM-dependent transcriptional repression leads to untimely early expression of central cell cycle regulators. The p53-p21-DREAM pathway represents one important means of cell cycle checkpoint activation by p53. By activating this pathway, p53 can downregulate transcription of genes controlled by DREAM. Here, we present a genome-wide ranked list of genes deregulated by HPV E7 expression and relate it to datasets of cell cycle genes and DREAM targets. We find that DREAM targets are generally deregulated after E7 expression. Furthermore, our analysis shows that p53-dependent downregulation of DREAM targets is abrogated when HPV E7 is expressed. Thus, p53 checkpoint control is impaired by HPV E7 independently of E6. In summary, our analysis reveals that disruption of DREAM through the HPV E7 oncoprotein upregulates most, if not all, cell cycle genes and impairs p53’s control of cell cycle checkpoints.

Abstract Background Focusing with a 4 π parabolic mirror allows for concentrating light from nearly the complete solid angle, whereas focusing with a single microscope objective limits the angle cone used for focusing to half solid angle at maximum. Increasing the solid angle by using deep parabolic mirrors comes at the cost of adding more complexity to the mirror’s fabrication process and might introduce errors that reduce the focusing quality. Methods To determine these errors, we experimentally examine the focusing properties of a 4 π parabolic mirror that was produced by single-point diamond turning. The properties are characterized with a single 174Yb + ion as a mobile point scatterer. The ion is trapped in a vacuum environment with a movable high optical access Paul trap. Results We demonstrate an effective focal spot size of 209 nm in lateral and 551 nm in axial direction. Such tight focusing allows us to build an efficient light-matter interface. Conclusion Our findings agree with numerical simulations incorporating a finite ion temperature and interferometrically measured wavefront aberrations induced by the parabolic mirror. We point at further technological improvements and discuss the general scope of applications of a 4 π parabolic mirror.

Subtype-specific human cardiomyocytes (CMs) are valuable for basic and applied research. Induction of cardiomyogenesis and enrichment of nodal-like CMs was described for mouse pluripotent stem cells (mPSCs) in response to 1-ethyl-2-benzimidazolinone (EBIO), a chemical modulator of small-/intermediate-conductance Ca2+-activated potassium channels (SKs 1–4). Investigating EBIO in human pluripotent stem cells (PSCs), we have applied three independent differentiation protocols of low to high cardiomyogenic efficiency. Equivalent to mPSCs, timed EBIO supplementation during hPSC differentiation resulted in dose-dependent enrichment of up to 80% CMs, including an increase in nodal- and atrial-like phenotypes. However, our study revealed extensive EBIO-triggered cell loss favoring cardiac progenitor preservation and, subsequently, CMs with shortened action potentials. Proliferative cells were generally more sensitive to EBIO, presumably via an SK-independent mechanism. Together, EBIO did not promote cardiogenic differentiation of PSCs, opposing previous findings, but triggered lineage-selective survival at a cardiac progenitor stage, which we propose as a pharmacological strategy to modulate CM subtype composition.

We investigate native nitrogen vacancy (NV) and silicon vacancy (SiV) color centers in a commercially available, heteroepitaxial, wafer-sized, mm thick, single-crystal diamond. We observe single, native NV centers with a density of roughly 1 NV per μm3 and moderate coherence time (T2 = 5 μs) embedded in an ensemble of SiV centers. Using low temperature luminescence of SiV centers as a probe, we prove the high crystalline quality of the diamond especially close to the growth surface, consistent with a reduced dislocation density. Using ion implantation and plasma etching, we verify the possibility to fabricate nanostructures with shallow color centers rendering our material promising for fabrication of nanoscale sensing devices. As this diamond is available in wafer-sizes up to 100 mm, it offers the opportunity to up-scale diamond-based device fabrication.

There is currently a large demand for aluminum components to measure the mechanical and thermal loads to which they are subjected. With structural health monitoring, components in planes, vehicles, or bridges can monitor critical loads and potentially prevent an impending fatigue failure. Externally attached sensors need a structural model to obtain knowledge of the mechanical load at the point of interest, whereas embedded sensors can be used for direct measurement at the point of interest. To produce an embedded sensor, which is automatically encapsulated against environmental influence, the sensor must be able to withstand the boundary conditions of the host component’s manufacturing process. This embedding process is particularly demanding in the case of casting. Previous work showed that silicon-based sensors have a high failure rate when embedded in cast aluminum parts and that using aluminum as a substrate is preferable under these circumstances. In the present paper, we present the fabrication process for the combination of a thick-film insulation and a thin-film strain gauge sensor, on such an aluminum substrate. The sensor is capable of withstanding high temperatures of at least 600 °C for over 20 min and a subsequent embedding in a gravity die casting process.

A implementação de tecnologias da informação na construção civil, como Virtual Design and Construction / Building Information Modeling (VDC/BIM) representam uma nova perspectiva para melhores práticas na AECO (Arquitetura; Engenharia; Construção; Operação). Contudo, no cenário brasileiro, estas práticas carecem de métodos para medição e análise de desempenho processual da informação na gestão do processo de projeto, demonstrando a deficiência da documentação de critérios e seleção de indicadores que contribuam para melhoria do setor da construção civil. Trata-se de uma pesquisa descritiva sobre a exploração do método VDC Scorecard e a adaptação à realidade brasileira, tendo por base pesquisa bibliográfica e documental existentes em bases de dados internacionais e nacionais explorando os seguintes aspectos: benefícios registrados; dificuldades na implementação; boas práticas na gestão do projeto. A pesquisa valida a importância dos métodos de desempenho que se adequem ao contexto nacional, viabilizando a integração e controle da informação entre agentes envolvidos num modelo colaborativo de edificação virtual e apropriação de dados por meio dos modelos de desempenho configurados através da análise contínua dos processos, da organização e do produto. Esta apropriação possibilita a documentação de um banco de dados de práticas eficientes para gestão de processos de projeto configurando o desempenho do modelo.

To harness the potential of human pluripotent stem cells (hPSCs), an abundant supply of their progenies is required. Here, hPSC expansion as matrix-independent aggregates in suspension culture was combined with cardiomyogenic differentiation using chemical Wnt pathway modulators. A multiwell screen was scaled up to stirred Erlenmeyer flasks and subsequently to tank bioreactors, applying controlled feeding strategies (batch and cyclic perfusion). Cardiomyogenesis was sensitive to the GSK3 inhibitor CHIR99021 concentration, whereas the aggregate size was no prevailing factor across culture platforms. However, in bioreactors, the pattern of aggregate formation in the expansion phase dominated subsequent differentiation. Global profiling revealed a culture-dependent expression of BMP agonists/antagonists, suggesting their decisive role in cell-fate determination. Furthermore, metallothionein was discovered as a potentially stress-related marker in hPSCs. In 100 ml bioreactors, the production of 40 million predominantly ventricular-like cardiomyocytes (up to 85% purity) was enabled that were directly applicable to bioartificial cardiac tissue formation.

We present a technique based on high resolution imaging to measure the absolute temperature and the heating rate of a single ion trapped at the focus of a deep parabolic mirror. We collect the fluorescence light scattered by the ion during laser cooling and image it onto a camera. Accounting for the size of the point-spread function and the magnification of the imaging system, we determine the spatial extent of the ion, from which we infer the mean phonon occupation number in the trap. Repeating such measurements and varying the power or the detuning of the cooling laser, we determine the heating rate induced by any kind of effect other than photon scattering. In contrast to other established schemes for measuring the heating rate, the ion is always maintained in a state of thermal equilibrium at temperatures close to the Doppler limit.

Substitution of degenerated dopaminergic (DA) neurons by intrastriatally transplanted ventral mesencephalon (VM)-derived progenitor cells has been shown to improve motor functions in parkinsonian patients and animal models, whereas investigations of electrophysiological properties of the grafted DA neurons have been rarely performed. Here we show electrophysiological properties of grafted VM progenitor cells at different time intervals up to 12 weeks after transplantation measured in acute brain slices using eGFP-Flag transfection to identify the graft. We were able to classify typical DA neurons according to the biphasic progression (voltage “sag”) to hyperpolarizing current injections. Two types of DA-like neurons were classified. Whereas type 1 neurons were characterized by delayed action potentials after hyperpolarization and irregular spontaneous firing, type 2 neurons displayed burst firing after hyperpolarization, spontaneous bursts, and regular firing. Comparison to identified DA neurons in vitro indicates a high integration of the intrastriatally grafted neurons, since in vitro cultures displayed regular firing spontaneously, whereas grafted identified DA neurons showed irregular firing. Additionally, type 1 and type 2 neurons exhibited a slight increase in the spontaneous firing frequency over time intervals after grafting, which might reflect a progressive integration of the grafted DA neurons. Our results provide evidence of the differentiation of grafted VM progenitor cells into mature integrated DA neurons, which are shown to replace the missing DA neurons functionally early after grafting.

Artificial nerve grafts are needed to reconstruct massive defects in the peripheral nervous system when autologous nerve grafts are not available in sufficient amounts. Nerve grafts containing Schwann cells display a suitable substrate for long-distance regeneration. We present here a comprehensive analysis of the in vivo effects of different isoforms of fibroblast growth factor-2 (FGF-2) on peripheral nerve regeneration across long gaps. FGF-2 isoforms were provided by grafted, genetically modified Schwann cells over-expressing 18-kDa-FGF-2 and 21-/23-kDa-FGF-2, respectively. Functional tests evaluated motor and sensory recovery. Additionally, morphometrical analyses of regenerated nerves were performed 3 and 6 months after grafting. Distinct regeneration promoting effects of the different FGF-2 isoforms were found. 18-kDa-FGF-2 mediated inhibitory effects on the grade of myelination of regenerating axons, whereas 21-/23-kDa-FGF-2 mediated early recovery of sensory functions and stimulation of long-distance myelination of regenerating axons. The results contribute to the development of new therapeutic strategies in peripheral nerve repair.

Scientific reasoning and scientific argumentation are highly valued outcomes of K-12 and higher education. In this article, we first review main topics and key findings of three different strands of research, namely research on the development of scientific reasoning, research on scientific argumentation, and research on approaches to support scientific reasoning and argumentation. Building on these findings, we outline current research deficits and address five aspects that exemplify where and how research on scientific reasoning and ar-gumentation needs to be expanded. In particular, we suggest to ground future research in a conceptual frame-work with three epistemic modes (advancing theory building about natural and social phenomena, artefact-centred scientific reasoning, and science-based reasoning in practice) and eight epistemic activities (problem identification, questioning, hypothesis generation, construction and redesign of artefacts, evidence generation, evidence evaluation, drawing conclusions as well as communicating and scrutinizing scientific reasoning and its results). We further propose addressing the domain specificities and domain generalities of scientific reasoning and approaches to its facilitation as well as investigating the role of epistemic emotions in scientific reasoning, the social context of SRA, and the influence of digital technologies on scientific reasoning and argumentation.

The microtubule-dependent molecular motor KIF23 (Kinesin family member 23) is one of two components of the centralspindlin complex assembled during late stages of mitosis. Formation of this complex is known as an essential step for cytokinesis. Here, we identified KIF23 as a new transcriptional target gene of the tumor suppressor protein p53. We showed that p53 reduces expression of KIF23 on the mRNA as well as the protein level in different cell types. Promoter reporter assays revealed that this repression results from downregulation of KIF23 promoter activity. CDK inhibitor p21(WAF1/CIP1) was shown to be necessary to mediate p53-dependent repression. Furthermore, we identified the highly conserved cell cycle genes homology region (CHR) in the KIF23 promoter to be strictly required for p53-dependent repression as well as for cell cycle-dependent expression of KIF23. Cell cycle- and p53-dependent regulation of KIF23 appeared to be controlled by differential binding of DREAM and MMB complexes to the CHR element. With this study, we describe a new mechanism for transcriptional regulation of KIF23. Considering the strongly supporting function of KIF23 in cytokinesis, its p53-dependent repression may contribute to the prevention of uncontrolled cell growth.

We study single silicon vacancy (SiV) centres in chemical vapour deposition (CVD) nanodiamonds on iridium as well as an ensemble of SiV centres in a high-quality, low-stress CVD diamond film by using temperature-dependent luminescence spectroscopy in the temperature range 5–295 K. We investigate in detail the temperature-dependent fine structure of the zero-phonon line (ZPL) of the SiV centres. The ZPL transition is affected by inhomogeneous as well as temperature-dependent homogeneous broadening and blue shifts by about 20 cm ^−1 upon cooling from room temperature to 5 K. We employ excitation power-dependent g ^(2) measurements to explore the temperature-dependent internal population dynamics of single SiV centres and infer mostly temperature-independent dynamics.