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A yeast expression plasmid was constructed containing a cardenolide biosynthetic module, referred to as CARD II, using the AssemblX toolkit, which enables the assembly of large DNA constructs. The genes cloned into the vector were (a) a Δ5‐3β‐hydroxysteroid dehydrogenase gene from Digitalis lanata, (b) a steroid Δ5‐isomerase gene from Comamonas testosteronii, (c) a mutated steroid‐5β‐reductase gene from Arabidopsis thaliana, and (d) a steroid 21‐hydroxylase gene from Mus musculus. A second plasmid bearing an ADR/ADX fusion gene from Bos taurus was also constructed. A Saccharomyces cerevisiae strain bearing these two plasmids was generated. This strain, termed “CARD II yeast”, was capable of producing 5β‐pregnane‐3β,21‐diol‐20‐one, a central intermediate in 5β‐cardenolide biosynthesis, starting from pregnenolone which was added to the culture medium. Using this approach, five consecutive steps in cardenolide biosynthesis were realized in baker's yeast.
The computation of such synthetic GFs is computationally and operationally demanding. As a consequence, the onthe-fly recalculation of synthetic GFs in each iteration of an optimisation is time-consuming and impractical. Therefore, the pre-calculation and efficient storage of synthetic GFs on a dense grid of source to receiver combinations enables the efficient lookup and utilisation of GFs in time-critical scenarios. We present a Python-based framework and toolkit - Pyrocko-GF - that enables the pre-calculation of synthetic GF stores, which are independent of their numerical calculation method and GF transfer function. The framework aids in the creation of such GF stores by interfacing a suite of established numerical forward modelling codes in seismology (computational back ends). So far, interfaces to back ends for layered Earth model cases have been provided; however, the architecture of Pyrocko-GF is designed to cover back ends for other geometries (e.g. full 3-D heterogeneous media) and other physical quantities (e.g. gravity, pressure, tilt). Therefore, Pyrocko-GF defines an extensible GF storage format suitable for a wide range of GF types, especially handling elasticity and wave propagation problems. The framework assists with visualisations, quality control, and the exchange of GF stores, which is supported through an online platform that provides many pre-calculated GF stores for local, regional, and global studies. The Pyrocko-GF toolkit comes with a well-documented application programming interface (API) for the Python programming language to efficiently facilitate forward modelling of geophysical processes, e.g. synthetic waveforms or static displacements for a wide range of source models.
The present study focuses on an innovative approach in measuring the mechanical oscillations of pre-loaded Achilles tendon by using Mechanotendography (MTG) during application of a short yet powerful mechanical pressure impact. This was applied on the forefoot from the plantar side in direction of dorsiflexion, while the subject stood on the ball of the forefoot on one leg. Participants with Achilles tendinopathy (AT; n = 10) were compared to healthy controls (Con; n = 10). Five trials were performed on each side of the body. For evaluation, two intervals after the impulse began (0-100ms; 30-100ms) were cut from the MTG and pressure raw signals. The intrapersonal variability between the five trials in both intervals were evaluated using the arithmetic mean and coefficient of variation of the mean correlation (Spearman rank correlation) and the normalized averaged mean distances, respectively. The AT-group showed a significantly reduced variability in MTG compared to the Con-group (from p = 0.006 to p = 0.028 for different parameters). The 95% confidence intervals (CI) of MTG results were disjoint, whereas the 95% CIs of the pressure signals were similar (p = 0.192 to p = 0.601). We suggest from this work that the variability of mechanical tendon oscillations could be an indicative parameter of an altered Achilles tendon functionality.
Cell-free protein synthesis (CFPS) based on eukaryotic Sf21 lysate is gaining interest among researchers due to its ability to handle the synthesis of complex human membrane proteins (MPs). Additionally Sf21 cell-free systems contain endogenous microsomal vesicles originally derived from the endoplasmic reticulum (ER). After CFPS, MPs will be translocated into the microsomal vesicles membranes present in the lysates. Thus microsomal membranes offer a natural environment for de novo synthesized MPs. Despite the advantage of synthesizing complex MPs with post translational modifications directly into the microsomal membranes without any additional solubilization supplements, batch based Sf21 cell-free synthesis suffers from low yields. The bottleneck for MPs in particular after the synthesis and incorporation into the microsomal membranes is to analyze their functionality. Apart from low yields of the synthesized MPs with batch based cell-free synthesis, the challenges arise in the form of cytoskeleton elements and peripheral endogenous proteins surrounding the microsomes which may impede the functional analysis of the synthesized proteins. So careful sample processing after the synthesis is particularly important for developing the appropriate functional assays. Here we demonstrate how MPs (native and batch synthesized) from ER derived microsomes can be processed for functional analysis by electrophysiology and radioactive uptake assay methods. Treatment of the microsomal membranes either with a sucrose washing step in the case of human serotonin transporter (hSERT) and sarco/endoplasmic reticulum Ca2+/ATPase (SERCA) pump or with mild detergents followed by the preparation of proteoliposomes in the case of the human voltage dependent anionic channel (hVDAC1) helps to analyze the functional properties of MPs.
Plants are in permanent contact with various microorganisms and are always impacted by them. To better understand the first steps of a plant’s recognition of soil-borne microorganisms, the early release of volatile organic compounds (VOCs) emitted from roots of Medicago truncatula in response to the symbiont Rhizophagus irregularis or the pathogenic oomycete Aphanomyces euteiches was analysed. More than 90 compounds were released from roots as detected by an untargeted gas chromatography-mass spectrometry approach. Principal component analyses clearly distinguished untreated roots from roots treated with either R. irregularis or A. euteiches. Several VOCs were found to be emitted specifically in response to each of the microorganisms. Limonene was specifically emitted from wild-type roots after contact with R. irregularis spores but not from roots of the mycorrhiza-deficient mutant does not make infections3. The application of limonene to mycorrhizal roots, however, did not affect the mycorrhization rate. Inoculation of roots with A. euteiches zoospores resulted in the specific emission of several sesquiterpenes, such as nerolidol, viridiflorol and nerolidol-epoxyacetate but application of nerolidol to zoospores of A. euteiches did not affect their vitality. Therefore, plants discriminate between different microorganisms at early stages of their interaction and respond differently to the level of root-emitted volatiles.
Understanding the development of spoken language in young children has become increasingly important for advancing basic theories of language acquisition and for clinical practice. However, such a goal requires refined measurements of speech articulation (e.g., from the tongue), which are difficult to obtain from young children. In recent years though, technological advances have allowed developmental researchers to make significant steps in that direction. For instance, movements of the tongue, an articulator that is essential for spoken language, can now be tracked and recorded in children with ultrasound imaging. This technique has opened novel research avenues in (a)typical language acquisition, enabling researchers to reliably capture what has long remained invisible in the speech of young children. Within this context, we have designed an experimental platform for the recording and the processing of kinematic data: SOLLAR (Sonographic and Optical Linguo-Labial Articulatory Recording system). The method has been tailored for children, but it is suitable for adults. In the present article, we introduce the recording environment developed to record over 100 children and 30 adults within SOLLAR. We then describe SOLLAR’s data processing framework, providing examples of data visualization and a summary of strengths and limitations.
URSA-PQ
(2020)
We present a highly flexible and portable instrument to perform pump-probe spectroscopy with an optical and an X-ray pulse in the gas phase. The so-called URSA-PQ (German for ‘Ultraschnelle Röntgenspektroskopie zur Abfrage der Photoenergiekonversion an Quantensystemen’, Engl. ‘ultrafast X-ray spectroscopy for probing photoenergy conversion in quantum systems’) instrument is equipped with a magnetic bottle electron spectrometer (MBES) and tools to characterize the spatial and temporal overlap of optical and X-ray laser pulses. Its adherence to the CAMP instrument dimensions allows for a wide range of sample sources as well as other spectrometers to be included in the setup. We present the main design and technical features of the instrument. The MBES performance was evaluated using Kr M4,5NN Auger lines using backfilled Kr gas, with an energy resolution ΔE/E ≅ 1/40 in the integrating operative mode. The time resolution of the setup at FLASH 2 FL 24 has been characterized with the help of an experiment on 2-thiouracil that is inserted via the instruments’ capillary oven. We find a time resolution of 190 fs using the molecular 2p photoline shift and attribute this to different origins in the UV-pump—the X-ray probe setup.
Social Media, Quo Vadis?
(2020)
Over the past two decades, social media have become a crucial and omnipresent cultural and economic phenomenon, which has seen platforms come and go and advance technologically. In this study, we explore the further development of social media regarding interactive technologies, platform development, relationships to news media, the activities of institutional and organizational users, and effects of social media on the individual and the society over the next five to ten years by conducting an international, two-stage Delphi study. Our results show that enhanced interaction on platforms, including virtual and augmented reality, somatosensory sense, and touch- and movement-based navigation are expected. AIs will interact with other social media users. Inactive user profiles will outnumber active ones. Platform providers will diversify into the WWW, e-commerce, edu-tech, fintechs, the automobile industry, and HR. They will change to a freemium business model and put more effort into combating cybercrime. Social media will become the predominant news distributor, but fake news will still be problematic. Firms will spend greater amounts of their budgets on social media advertising, and schools, politicians, and the medical sector will increase their social media engagement. Social media use will increasingly lead to individuals’ psychic issues. Society will benefit from economic growth and new jobs, increased political interest, democratic progress, and education due to social media. However, censorship and the energy consumption of platform operators might rise.
This study aimed to investigate the relationship between the acute to chronic workload ratio (ACWR), based upon participant session rating of perceived exertion (sRPE), using two models [(1) rolling averages (ACWRRA); and (2) exponentially weighted moving averages (ACWREWMA)] and the injury rate in young male team soccer players aged 17.1 ± 0.7 years during a competitive mesocycle. Twenty-two players were enrolled in this study and performed four training sessions per week with 2 days of recovery and 1 match day per week. During each training session and each weekly match, training time and sRPE were recorded. In addition, training impulse (TRIMP), monotony, and strain were subsequently calculated. The rate of injury was recorded for each soccer player over a period of 4 weeks (i.e., 28 days) using a daily questionnaire. The results showed that over the course of the study, the number of non-contact injuries was significantly higher than that for contact injuries (2.5 vs. 0.5, p = 0.01). There were also significant positive correlations between sRPE and training time (r = 0.411, p = 0.039), ACWRRA (r = 0.47, p = 0.049), and ACWREWMA (r = 0.51, p = 0.038). In addition, small-to-medium correlations were detected between ACWR and non-contact injury occurrence (ACWRRA, r = 0.31, p = 0.05; ACWREWMA, r = 0.53, p = 0.03). Explained variance (r 2) for non-contact injury was significantly greater using the ACWREWMA model (ranging between 21 and 52%) compared with ACWRRA (ranging between 17 and 39%). In conclusion, the results of this study showed that the ACWREWMA model is more sensitive than ACWRRA to identify non-contact injury occurrence in male team soccer players during a short period in the competitive season.
The Jewish family has been the subject of much admiration and analysis, criticism and myth-making, not just but especially in modern times. As a field of inquiry, its place is at the intersection – or in the shadow – of the great topics in Jewish Studies and its contributing disciplines. Among them are the modernization and privatization of Judaism and Jewish life; integration and distinctiveness of Jews as individuals and as a group; gender roles and education. These and related questions have been the focus of modern Jewish family research, which took shape as a discipline in the 1910s.
This issue of PaRDeS traces the origins of academic Jewish family research and takes stock of its development over a century, with its ruptures that have added to the importance of familial roots and continuities. A special section retrieves the founder of the field, Arthur Czellitzer (1871–1943), his biography and work from oblivion and places him in the context of early 20th-century science and Jewish life.
The articles on current questions of Jewish family history reflect the topic’s potential for shedding new light on key questions in Jewish Studies past and present. Their thematic range – from 13th-century Yiddish Arthurian romances via family-based business practices in 19th-century Hungary and Germany, to concepts of Jewish parenthood in Imperial Russia – illustrates the broad interest in Jewish family research as a paradigm for early modern and modern Jewish Studies.