DNA replication machinery: Insights from in vitro single-molecule approaches.

The replisome is the multiprotein molecular machinery that replicates DNA. The replisome components work in precise coordination to unwind the double helix of the DNA and replicate the two strands simultaneously. The study of DNA replication using in vitro single-molecule approaches provides a novel quantitative understanding of the dynamics and mechanical principles that govern the operation of the replisome and its components. 'Classical' ensemble-averaging methods cannot obtain this information. Here we describe the main findings obtained with in vitro single-molecule methods on the performance of individual replisome components and reconstituted prokaryotic and eukaryotic replisomes. The emerging picture from these studies is that of stochastic, versatile and highly dynamic replisome machinery in which transient protein-protein and protein-DNA associations are responsible for robust DNA replication.

Anion Exchange Membranes with Dynamic Redox-Responsive Properties.

Redox-responsive anion exchange membranes were developed using photoinitiated free-radical polymerization and reversible oxidation and reduction of viologen. The membranes were formulated using poly(ethylene glycol diacrylate) and diurethane dimethacrylate oligomers, dipentaerythritol penta-/hexa-acrylate cross-linker, photoinitiators, and 4-vinylbenzyl chloride as precursors for functionalization. In the membrane, 4,4'-bipyridine reacted with the 4-vinylbenzyl chloride residues, and subsequently, unreacted amines were methylated with iodomethane to obtain viologen as both the ion carrier and redox-responsive group. Upon oxidation, viologen supports two cations, where the reduced form only contains one cation. Thus, the redox responsiveness changed the membrane ionicity by a factor of 2. The area-specific resistance of the membranes in the oxidized, +2, state was lower than in the reduced, +1, state. The resistance increased between 40.6 ± 0.1 and 111.6 ± 0.1%, depending on membrane thickness, with the most significant increment being a resistance change from 4.88 × 10-4 Ω m2 in the oxidized state to 1.03 × 10-3 Ω m2 in the reduced state. Membrane permselectivity in the reduced, +1, state was between 15.9 ± 0.1 and 26.5 ± 0.01% lower than in the oxidized, +2, state, with no change in water uptake, spanning an average of 0.87 ± 0.02 in the oxidized state to an average of 0.7 ± 0.01 in the reduced state. Upon reduction, membrane ion-exchange capacity decreases, increasing ionic resistance and decreasing membrane permselectivity due to a reduction in fixed charge concentration without a measurable change in water uptake. This trend is not generally observed for ion-exchange membranes and explains that the changes in transport properties result from changes in ionicity, not water uptake or domain size. The reversibility and stability of the stimuli responsiveness were confirmed by the absence of transport property changes after redox cycling.

Duckweed as an Agricultural Amendment: Nitrogen Mineralization, Leaching, and Sorghum Uptake.

Excessive N and P in surface waters can promote eutrophication (algae-dominated, low-O waters), which decreases water quality and aquatic life. Duckweed (Lemnaceae), a floating aquatic plant, rapidly absorbs N and P from water and its composition shows strong potential as a soil amendment. Therefore, it may be used to transfer N and P from eutrophic water bodies to agricultural fields. In this work, dried duckweed was incorporated into agricultural soil in microcosm, column, and field tests to evaluate biological N cycling, nutrient retention, and crop yield compared with compost, diammonium phosphate (DAP), and an amendment-free control. In microcosm tests, 25 ± 13% of duckweed N was mineralized, providing on average less mineral N than DAP (107 ± 21%), but more than compost (11 ± 12%). In columns, duckweed treatments leached only 2% of the N added, significantly less than DAP, which leached 60% of its N. Compared with the control, DAP leached significantly more phosphate (78%), whereas duckweed and compost treatments leached less (56 and 27%, respectively). Crop yield, as well as runoff N and P, were measured in field tests growing forage sorghum [ (L.) Moench.]. Although less total N was applied to duckweed plots than to DAP plots (75 vs. 130 kg ha, respectively), duckweed was found to retain 30% more total mineral N in a tilled agricultural field than DAP, while supporting a comparable yield. These tests indicate that duckweed may provide a sustainable source of N and P for agriculture.

Resistance and Permselectivity of 3D-Printed Micropatterned Anion-Exchange Membranes.

It has been demonstrated that a micropatterned surface can decrease the resistance of anion-exchange membranes (AEMs) and can induce desirable flow properties in devices, such as mixing. Previously, a model that related the resistance of flat and patterned membranes with the same equivalent thickness was proposed, which used the patterned area and thickness ratio of the features to describe the membrane resistance. Here, we explored the validity of the parallel resistance model for a variety of membrane surface designs and area ratios. We demonstrated that the model can predict the resistance of a wide range of patterned AEMs. We showed that the resistance is independent of the spatial ordering of the design by examining random patterns, which is relevant for applications that require, for example, increased mixing in multilayered devices. Some experimental values of resistance obtained for patterned membranes presented deviations from the model. Scanning electron microscopy (SEM) images of the patterned membranes revealed resolution variations and pattern replication errors due to the stereolithographic process. A geometric correction of the target ratios improved the fit of the modeled data to the experimental values, showing that light bleeding during curing was a source of error. Two additional experimental factors were not accounted for in the model: a distinct interface between the bottom and top layer and overcuring of the bottom layer during successive steps. These sources of error were investigated by examining the resistance of single- and double-layered membranes, as well as single-layer membranes with different curing times. The differences obtained in the resistances for control samples demonstrated that both the interface and the overcuring influenced the resistance of the membrane. The results obtained in this study enlighten the discussion relating membrane-surface morphology and transport properties, as well as the optimization of 3D-printed membranes using a stereolithography process.

Randomized Controlled Trial Examining Effects of Web-Based Information on Patient Satisfaction and Image Quality in 18F-FDG PET/CT Examinations.

Our aim was to compare the effect that having access, versus not having access, to web-based patient information on 18F-FDG PET/CT has on image quality and on patient satisfaction with their care during and knowledge about the examination, as well as to explore whether patients utilized and were satisfied with the web-based information. Methods: We recruited 148 patients between October 2015 and December 2016 and randomly assigned them to a standard-care group or an intervention group. Both groups received standard information about the 18F-FDG PET/CT examination, but the intervention group also received access to web-based information. A questionnaire was used to evaluate patient satisfaction with, knowledge about, and discomfort during the examination, and a masked assessment of image quality was conducted. Results: Overall satisfaction was high in both groups. The lowest satisfaction was with information about how the patients would receive the results of the examination. More patients in the intervention group than in the standard-care group knew how the 18F-FDG PET/CT examination would be conducted. Descriptive data suggest that image quality was slightly better in the intervention group than in the standard-care group, but none of the outcomes significantly differed between the groups. However, several obstacles were encountered during recruitment that led to insufficient power to detect differences. Also, only 54 of 75 patients (72%) in the intervention group utilized the web-based information. However, those who did utilize the information were satisfied with it and found it helpful. Conclusion: The effects of web-based information need to be investigated in a larger sample of patients. Having access to improved information before undergoing 18F-FDG PET/CT may help patients prepare for and undergo the examination. It may also improve image quality. However, this possibility needs to be investigated using image quality as the primary outcome. The results may be used to improve patient information and care and thereby optimize the 18F-FDG PET/CT procedure.

INVO procedure: minimally invasive IVF as an alternative treatment option for infertile couples.

Intravaginal culture (IVC), also called INVO (intravaginal culture of oocytes), is an assisted reproduction procedure where oocyte fertilization and early embryo development are carried out within a gas permeable air-free plastic device, placed into the maternal vaginal cavity for incubation. In the present study we assessed the outcome of the INVO procedure, using the recently designed INVOcell device, in combination with a mild ovarian stimulation protocol. A total of 125 cycles were performed. On average 6.5 oocytes per cycle were retrieved, and a mean of 4.2 were placed per INVOcell device. The cleavage rate obtained after the INVO culture was 63%. The procedure yielded 40%, 31.2%, and 24% of clinical pregnancy, live birth, and single live birth rates per cycle, respectively. Our results suggest that the INVO procedure is an effective alternative treatment option in assisted reproduction that shows comparable results to those reported for existing IVF techniques.