A Novel Biomimetic Tool for Assessing Vitamin K Status Based on Molecularly Imprinted Polymers

Vitamin K was originally discovered as a cofactor required to activate clotting factors and has recently been shown to play a key role in the regulation of soft tissue calcification. This property of vitamin K has led to an increased interest in novel methods for accurate vitamin K detection. Molecularly Imprinted Polymers (MIPs) could offer a solution, as they have been used as synthetic receptors in a large variety of biomimetic sensors for the detection of similar molecules over the past few decades, because of their robust nature and remarkable selectivity. In this article, the authors introduce a novel imprinting approach to create a MIP that is able to selectively rebind vitamin K 1. As the native structure of the vitamin does not allow for imprinting, an alternative imprinting strategy was developed, using the synthetic compound menadione (vitamin K 3) as a template. Target rebinding was analyzed by means of UV-visible (UV-VIS) spectroscopy and two custom-made thermal readout techniques. This analysis reveals that the MIP-based sensor reacts to an increasing concentration of both menadione and vitamin K 1. The Limit of Detection (LoD) for both compounds was established at 700 nM for the Heat Transfer Method (HTM), while the optimized readout approach, Thermal Wave Transport Analysis (TWTA), displayed an increased sensitivity with a LoD of 200 nM. The sensor seems to react to a lesser extent to Vitamin E, the analogue under study. To further demonstrate its potential application in biochemical research, the sensor was used to measure the absorption of vitamin K in blood serum after taking vitamin K supplements. By employing a gradual enrichment strategy, the sensor was able to detect the difference between baseline and peak absorption samples and was able to quantify the vitamin K concentration in good agreement with a validation experiment using High-Performance Liquid Chromatography (HPLC). In this way, the authors provide a first proof of principle for a low-cost, straightforward, and label-free vitamin K sensor

Materials and Textile Architecture Analyses for Mechanical Counter-Pressure Space Suits using Active Materials

Mechanical counter-pressure (MCP) space suits have the potential to improve the mobility of astronauts as they conduct planetary exploration activities. MCP suits differ from traditional gas-pressurized space suits by applying surface pressure to the wearer using tight-fitting materials rather than pressurized gas, and represent a fundamental change in space suit design. However, the underlying technologies required to provide uniform compression in a MCP garment at sufficient pressures for space exploration have not yet been perfected, and donning and doffing a MCP suit remains a significant challenge. This research effort focuses on the novel use of active material technologies to produce a garment with controllable compression capabilities (up to 30 kPa) to address these problems. We provide a comparative study of active materials and textile architectures for MCP applications; concept active material compression textiles to be developed and tested based on these analyses; and preliminary biaxial braid compression garment modeling results.United States. National Aeronautics and Space Administration (OCT Space Technology Research Fellowship Grant NNX11AM62H)MIT-Portugal Progra

Synthesis Paper: Targeted Livestock Grazing: Prescription for Healthy Rangelands

Targeted livestock grazing is a proven tool for manipulating rangeland vegetation, and current knowledge about targeted livestock grazing is extensive and expanding rapidly. Targeted grazing prescriptions optimize the timing, frequency, intensity, and selectivity of grazing (or browsing) in combinations that purposely exert grazing/browsing pressure on specific plant species or portions of the landscape. Targeted grazing differs from traditional grazing management in that the goal of targeted grazing is to apply defoliation or trampling to achieve specific vegetation management objectives, whereas the goal of traditional livestock grazing management is generally the production of livestock commodities. A shared aim of targeted livestock grazing and traditional grazing management is to sustain healthy soils, flora, fauna, and water resources that, in turn, can sustain natural ecological processes (e.g., nutrient cycle, water cycle, energy flow). Targeted grazing prescriptions integrate knowledge of plant ecology, livestock nutrition, and livestock foraging behavior. Livestock can be focused on target areas through fencing, herding, or supplement placement. Although practices can be developed to minimize the impact of toxins contained in target plants, the welfare of the animals used in targeted grazing must be a priority. Monitoring is needed to determine if targeted grazing is successful and to refine techniques to improve efficacy and efficiency. Examples of previous research studies and approaches are presented to highlight the ecological benefits that can be achieved when targeted grazing is applied properly. These cases include ways to suppress invasive plants and ways to enhance wildlife habitat and biodiversity. Future research should address the potential to select more adapted and effective livestock for targeted grazing and the associated animal welfare concerns with this practice. Targeted livestock grazing provides land managers a viable alternative to mechanical, chemical, and prescribed fire treatments to manipulate rangeland vegetation

A Human Neurovascular Unit On-a-Chip

Protection of the central nervous system (CNS) and cerebral homeostasis depend upon the blood–brain barrier (BBB) functions and permeability. BBB restrictive permeability hinders drug delivery for the treatment of several neurodegenerative diseases and brain tumors. Several in vivo animal models and in vitro systems have been developed to understand the BBB complex mechanisms and aid in the design of improved therapeutic strategies. However, there are still many limitations that should be addressed to achieve the structural and chemical environment of a human BBB. We developed a microfluidic-based model of the neurovascular unit. A monolayer of human cerebral endothelial cells (hCMEC-D3) was grown and cocultured with human brain microvascular pericytes (hBMVPC), and human induced pluripotent stem cells differentiated into astrocytes (hiPSC-AC) and neurons (hiPSC-N). To visualize the physiological morphology of each cell type, we used fluorescent cell-specific markers and confocal microscopy. Permeation of fluorescent solutes with different molecular weights was measured to demonstrate that the developed BBB was selectively permeable as a functional barrier

Topographical Vacuum Sealing of 3D-Printed Multiplanar Microfluidic Structures

We demonstrate a novel way of creating three-dimensional microfluidic channels capable of following complex topographies. To this end, substrates with open channels and different geometries were 3D-printed, and the open channels were consecutively closed with a thermoplastic using a low-resolution vacuum-forming approach. This process allows the sealing of channels that are located on the surface of complex multiplanar topographies, as the thermoplastic aligns with the surface-shape (the macrostructure) of the substrate, while the microchannels remain mostly free of thermoplastic as their small channel size resists thermoplastic inflow. This new process was analyzed for its capability to consistently close different substrate geometries, which showed reliable sealing of angles >90°. Furthermore, the thermoplastic intrusion into channels of different widths was quantified, showing a linear effect of channel width and percentage of thermoplastic intrusion; ranging from 43.76% for large channels with 2 mm width to only 5.33% for channels with 500 µm channel width. The challenging sealing of substrate ‘valleys’, which are created when two large protrusions are adjacent to each other, was investigated and the correlation between protrusion distance and height is shown. Lastly, we present three application examples: a serpentine mixer with channels spun around a cuboid, increasing the usable surface area; a cuvette-inspired flow cell for a 2-MXP biosensor based on molecular imprinted polymers, fitting inside a standard UV/Vis-Spectrophotometer; and an adapter system that can be manufactured by one-sided injection molding and is self-sealed before usage. These examples demonstrate how this novel technology can be used to easily adapt microfluidic circuits for application in biosensor platforms

Cost-effective, scalable and smartphone-controlled 3D-Printed syringe pump - From lab bench to point of care biosensing applications

Laboratories all over the word use syringe pumps every day for a multitude of purposes. The market of syringe pumps is limited, as it does not consider the broad range of specifications required by different researchers. In this work, we present a 3D printed syringe pump designed to be affordable, customizable, and extremely user friendly while still maintaining reliability and precision. The pump, thanks to its flexible design and smartphone-controlled interface, can be used in educational settings as well as in biological and chemical laboratories. The presented syringe-pump is used in this work to run a light catalyzed polymerization of butyl methacrylate using visible light, in a continuous flow setup

Biomimetic sensing of Escherichia coli at the solid-liquid interface: From surface-imprinted polymer synthesis toward real sample sensing in food safety

The incorporation of imprinted polymers (IP) into sensing devices has enabled the detection of a wide range of analytes going from molecules to whole cells. Biomimetic platforms for bacteria recognition possess the potential of being a low-cost and on-site testing technology for applications in food safety, where the accurate detection of pathogens is crucial for preventing disease. This work presents the optimization of a label-free thermal IP-based sensor for the detection of E. coli, a microorganism indicator of fecal contamination in drinking water and milk. By modifying the functional monomers employed in the synthesis of imprinted polyurethanes, the production process was sped up by a factor four. The receptor layers, which exhibited enhanced imprint surface-coverage, yielded in bacteria quantification at concentrations in the range of 1000 CFU/mL, an improvement of a full order of magnitude for the thermal sensor. Benchmarking of the results was performed with a commercial impedance analyzer, demonstrating the suitability of the receptors for electrochemical analysis. Furthermore, the results presented assess the selectivity of the thermal device against S. aureus. Finally, a proof of concept of the sensor in milk as complex sample is presented for the first time, confirming that the dynamic range and sensitivity of the device will suffice to detect fecal contamination of milk samples in resource-deprived settings