Shaping Giant Membrane Vesicles in 3D-Printed Protein Hydrogel Cages

Giant unilamellar phospholipid vesicles are attractive starting points for constructing minimal living cells from the bottom-up. Their membranes are compatible with many physiologically functional modules and act as selective barriers, while retaining a high morphological flexibility. However, their spherical shape renders them rather inappropriate to study phenomena that are based on distinct cell shape and polarity, such as cell division. Here, a microscale device based on 3D printed protein hydrogel is introduced to induce pH-stimulated reversible shape changes in trapped vesicles without compromising their free-standing membranes. Deformations of spheres to at least twice their aspect ratio, but also toward unusual quadratic or triangular shapes can be accomplished. Mechanical force induced by the cages to phase-separated membrane vesicles can lead to spontaneous shape deformations, from the recurrent formation of dumbbells with curved necks between domains to full budding of membrane domains as separate vesicles. Moreover, shape-tunable vesicles are particularly desirable when reconstituting geometry-sensitive protein networks, such as reaction-diffusion systems. In particular, vesicle shape changes allow to switch between different modes of self-organized protein oscillations within, and thus, to influence reaction networks directly by external mechanical cues

Molecular dynamics force probe simulations of antibody/antigen unbinding: Entropic control and non-additivity of unbinding forces.

Unbinding of a spin-labeled dinitrophenyl (DNP) hapten from the monoclonal antibody AN02 Fab fragment has been studied by force probe molecular dynamics (FPMD) simulations. In our nanosecond simulations, unbinding was enforced by pulling the hapten molecule out of the binding pocket. Detailed inspection of the FPMD trajectories revealed a large heterogeneity of enforced unbinding pathways and a correspondingly large flexibility of the binding pocket region, which exhibited induced fit motions. Principal component analyses were used to estimate the resulting entropic contribution of ∼6 kcal/mol to the AN02/DNP-hapten bond. This large contribution may explain the surprisingly large effect on binding kinetics found for mutation sites that are not directly involved in binding. We propose that such “entropic control” optimizes the binding kinetics of antibodies. Additional FPMD simulations of two point mutants in the light chain, Y33F and I96K, provided further support for a large flexibility of the binding pocket. Unbinding forces were found to be unchanged for these two mutants. Structural analysis of the FPMD simulations suggests that, in contrast to free energies of unbinding, the effect of mutations on unbinding forces is generally nonadditive

Membrane-coated 3D architectures for bottom-up synthetic biology dagger

One of the great challenges of bottom-up synthetic biology is to recreate the cellular geometry and surface functionality required for biological reactions. Of particular interest are lipid membrane interfaces where many protein functions take place. However, cellular 3D geometries are often complex, and custom-shaping stable lipid membranes on relevant spatial scales in the micrometer range has been hard to accomplish reproducibly. Here, we use two-photon direct laser writing to 3D print microenvironments with length scales relevant to cellular processes and reactions. We formed lipid bilayers on the surfaces of these printed structures, and we evaluated multiple combinatorial scenarios, where physiologically relevant membrane compositions were generated on several different polymer surfaces. Functional dynamic protein systems were reconstituted in vitro and their self-organization was observed in response to the 3D geometry. This method proves very useful to template biological membranes with an additional spatial dimension, and thus allows a better understanding of protein function in relation to the complex morphology of cells and organelles.We also thank the Biochemistry Core Facility of the Max Planck Institute of Biochemistry for assistance with protein purification, and the Imaging Core Facility of the same institution for assistance on the 4D image visualisation

Parent-Child Interaction Therapy: Theory and Research to Practice

This chapter will focus on the theory behind and research on Parent-Child Interaction Therapy (PCIT), a treatment developed for young children with disruptive behavior problems. We will describe and provide details about PCIT, which is based on both attachment and social learning models, and incorporates an innovative approach to treatment in which therapists coach caregivers “live” via a wireless headset while each caregiver interacts with their child. In addition, we will review research that has examined PCIT with a variety of diverse populations (e.g., children with developmental delay, physical abuse histories, anxiety and depression, and children from underrepresented racial and ethnic minority families), settings (e.g., clinic, home, school) and formats (e.g., individual, group, intensive). Finally, we will present a case study of PCIT with a child younger than 2 years to demonstrate the effectiveness of PCIT and highlight some common challenges and pitfalls that clinicians may face in clinical practice

Concatenated permutation block codes based on set partitioning for substitution and deletion error-control

A new class of permutation codes is presented where, instead of considering one permutation as a codeword, codewords consist of a sequence of permutations. The advantage of using permutations, i.e. their favourable symbol diversity properties, is preserved. Additionally, using sequences of permutations as codewords, code rates close to the optimum rate can be achieved. Firstly, the complete set of permutations is divided into subsets by using set partitioning. Binary data is then mapped to permutations from these subsets. These permutations, together with a parity permutation, will form the codeword. Two constructions will be presented: one capable of detecting and correcting substitution errors and the other capable of detecting and correcting either substitution or deletion errors

Effects of Extended Grape Ripening With or Without Must and Wine Alcohol Manipulations on Cabernet Sauvignon Wine Sensory Characteristics

This study attempts to clarify the consequences for wine flavour that result from harvesting fruit atdifferent maturities. The grapes were harvested from a single vineyard in Paso Robles, and the samplesspanned maturity levels from what would be considered early harvest (about 21 °Brix) to late harvest(about 30 °Brix). The wines made from these grapes were analysed using descriptive analysis to investigatethe relationships between fruit maturity and wine sensory attributes. In addition, musts and/or wines werechaptalised and/or fortified or watered back to determine the effect of these manipulations on wine sensoryproperties. This research showed that the sensory attributes of wines made from grapes at different stagesof maturation, from about 20 to 30 °Brix, varied in a systematic fashion. Specifically, the wines made fromthe grapes with a lower Brix were more sour and had more fresh vegetative flavours, while the wines madefrom the fruit with a high Brix were more hot and bitter and in some cases had more dark fruit flavoursand sweetness. Fortifying wines made from lower Brix musts changed the perceptions of the wine sensoryprofiles more than chaptalising the musts. On the other hand, adding water to higher °Brix musts to mimic24 °Brix musts resulted in wines with similar sensory profiles to wines made from grapes picked at a sugarcontent of close to 24 °Brix. This study shows that wine sensory attributes differ more when grapes arepicked early in ripening rather than after 24 °Brix

Long-term analysis of carbon dioxide and methane column-averaged mole fractions retrieved from SCIAMACHY

Carbon dioxide (CO<sub>2</sub>) and methane (CH<sub>4</sub>) are the two most important anthropogenic greenhouse gases contributing to global climate change. SCIAMACHY onboard ENVISAT (launch 2002) was the first and is now with TANSO onboard GOSAT (launch 2009) one of only two satellite instruments currently in space whose measurements are sensitive to CO<sub>2</sub> and CH<sub>4</sub> concentration changes in the lowest atmospheric layers where the variability due to sources and sinks is largest. <br><br> We present long-term SCIAMACHY retrievals (2003–2009) of column-averaged dry air mole fractions of both gases (denoted XCO<sub>2</sub> and XCH<sub>4</sub>) derived from absorption bands in the near-infrared/shortwave-infrared (NIR/SWIR) spectral region focusing on large-scale features. The results are obtained using an upgraded version (v2) of the retrieval algorithm WFM-DOAS including several improvements, while simultaneously maintaining its high processing speed. The retrieved mole fractions are compared to global model simulations (CarbonTracker XCO<sub>2</sub> and TM5 XCH<sub>4</sub>) being optimised by assimilating highly accurate surface measurements from the NOAA/ESRL network and taking the SCIAMACHY averaging kernels into account. The comparisons address seasonal variations and long-term characteristics. <br><br> The steady increase of atmospheric carbon dioxide primarily caused by the burning of fossil fuels can be clearly observed with SCIAMACHY globally. The retrieved global annual mean XCO<sub>2</sub> increase agrees with CarbonTracker within the error bars (1.80±0.13 ppm yr<sup>−1</sup> compared to 1.81±0.09 ppm yr<sup>−1</sup>). The amplitude of the XCO<sub>2</sub> seasonal cycle as retrieved by SCIAMACHY, which is 4.3±0.2 ppm for the Northern Hemisphere and 1.4±0.2 ppm for the Southern Hemisphere, is on average about 1 ppm larger than for CarbonTracker. <br><br> An investigation of the boreal forest carbon uptake during the growing season via the analysis of longitudinal gradients shows good agreement between SCIAMACHY and CarbonTracker concerning the overall magnitude of the gradients and their annual variations. The analysis includes a discussion of the relative uptake strengths of the Russian and North American boreal forest regions. <br><br> The retrieved XCH<sub>4</sub> results show that after years of stability, atmospheric methane has started to rise again in recent years which is consistent with surface measurements. The largest increase is observed for the tropics and northern mid- and high-latitudes amounting to about 7.5±1.5 ppb yr<sup>−1</sup> since 2007. Due care has been exercised to minimise the influence of detector degradation on the quantitative estimate of this anomaly

The effectiveness of interventions in workplace health promotion as to maintain the working capacity of health care personal

Background: The increasing proportion of elderly people with respective care requirements and within the total population stands against aging personnel and staff reduction in the field of health care where employees are exposed to high load factors. Health promotion interventions may be a possibility to improve work situations and behavior. Methods: A systematic literature search is conducted in 32 databases limited to English and German publications since 1990. Moreover, internet-searches are performed and the reference lists of identified articles are scanned. The selection of literature was done by two reviewers independently according to inclusion and exclusion criteria. Data extraction and tables of evidence are verified by a second expert just like the assessment of risk of bias by means of the Cochrane Collaboration’s tool. Results: We identified eleven intervention studies and two systematic reviews. There were three randomized controlled trials (RCT) and one controlled trial without randomization (CCT) on the improvement of physical health, four RCT and two CCT on the improvement of psychological health and one RCT on both. Study duration ranged from four weeks to two years and the number of participants included from 20 to 345, with a median of 56. Interventions and populations were predominantly heterogeneous. In three studies intervention for the improvement of physical health resulted in less complaints and increased strength and flexibility with statistically significant differences between groups. Regarding psychological health interventions lead to significantly decreased intake of analgesics, better stress management, coping with workload, communication skills and advanced training. Discussion: Taking into consideration the small to very small sample sizes, other methodological flaws like a high potential of bias and poor quality of reporting the validity of the results has to be considered as limited. Due to the heterogeneity of health interventions, study populations with differing job specializations and different lengths of study durations and follow-up periods, the comparison of results would not make sense. Conclusions: Further research is necessary with larger sample sizes, with a sufficient study duration and follow-up, with a lower risk of bias, by considering of relevant quality criteria and with better reporting in publications