Design Rule for Constructing Buckling-Free Polymeric Stencil with Microdot Apertures.
Ontology highlight
ABSTRACT:
SUBMITTER: Kim M
PROVIDER: S-EPMC8709208 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:We report on a simple and effective process that allows fabricating polymeric dual-scale nanoimprinting molds. The key for the process is the use of a thin flexible SU-8 stencil membrane, which was fabricated by either photolithography or thermal nanoimprint lithography (NIL). The stencil membrane with microscale pores was assembled into a nanopatterned substrate, producing a dual-scale structure. The assembled structure was used as a template to produce polymeric imprinting molds via UV-NIL. With this method, we demonstrated dual-scale nanoimprint molds having nano-pillars of 251 nm diameter and 146 nm high on top of microscale square protrusions of 5 μm wide and 3.6 μm high. The resin mold with the dual-scale structure was successfully used to produce a freestanding membrane with dual-scale perforated pores via UV-NIL. After metal coating and integrated into microfluidic devices, this freestanding membrane can potentially be used as a substrate for surface plasmon resonance sensors.
| S-EPMC6474418 | biostudies-literature
Project description:When pollen grains become exposed to the environment, they rapidly desiccate. To protect themselves until rehydration, the grains undergo characteristic infolding with the help of special structures in the grain wall-apertures-where the otherwise thick exine shell is absent or reduced in thickness. Recent theoretical studies have highlighted the importance of apertures for the elastic response and the folding of the grain. Experimental observations show that different pollen grains sharing the same number and type of apertures can nonetheless fold in quite diverse fashions. Using the thin-shell theory of elasticity, we show how both the absolute elastic properties of the pollen wall and the relative elastic differences between the exine wall and the apertures play an important role in determining pollen folding upon desiccation. Focusing primarily on colpate pollen, we delineate the regions of pollen elastic parameters where desiccation leads to a regular, complete closing of all apertures and thus to an infolding which protects the grain against water loss. Phase diagrams of pollen folding pathways indicate that an increase in the number of apertures leads to a reduction of the region of elastic parameters where the apertures close in a regular fashion. The infolding also depends on the details of the aperture shape and size, and our study explains how the features of the mechanical design of apertures influence the pollen folding patterns. Understanding the mechanical principles behind pollen folding pathways should also prove useful for the design of the elastic response of artificial inhomogeneous shells.
| S-EPMC7604445 | biostudies-literature
Project description:The self-assembled bilayer lipid membrane (BLM) is the basic component of the cell membrane. The reconstitution of ion channel proteins in artificially formed BLMs represents a well-defined system for the functional analysis of ion channels and screening the effects of drugs that act on them. However, because BLMs are unstable, this limits the experimental throughput of BLM reconstitution systems. Here we report on the formation of mechanically stable solvent-free BLMs in microfabricated apertures with defined nano- and micro-tapered edge structures. The role of such nano- and micro-tapered structures on the stability of the BLMs was also investigated. Finally, this BLM system was combined with a cell-free synthesized human ether-a-go-go-related gene channel, a cardiac potassium channel whose relation to arrhythmic side effects following drug treatment is well recognized. Such stable BLMs as these, when combined with a cell-free system, represent a potential platform for screening the effects of drugs that act on various ion-channel genotypes.
| S-EPMC5735097 | biostudies-literature
Project description:BackgroundShort hairpin RNA (shRNA) encoded within an expression vector has proven an effective means of harnessing the RNA interference (RNAi) pathway in mammalian cells. A survey of the literature revealed that shRNA vector construction can be hindered by high mutation rates and the ensuing sequencing is often problematic. Current options for constructing shRNA vectors include the use of annealed complementary oligonucleotides (74 % of surveyed studies), a PCR approach using hairpin containing primers (22 %) and primer extension of hairpin templates (4 %).ResultsWe considered primer extension the most attractive method in terms of cost. However, in initial experiments we encountered a mutation frequency of 50 % compared to a reported 20-40 % for other strategies. By modifying the technique to be an isothermal reaction using the DNA polymerase Phi29, we reduced the error rate to 10 %, making primer extension the most efficient and cost-effective approach tested. We also found that inclusion of a restriction site in the loop could be exploited for confirming construct integrity by automated sequencing, while maintaining intended gene suppression.ConclusionIn this study we detail simple improvements for constructing and sequencing shRNA that overcome current limitations. We also compare the advantages of our solutions against proposed alternatives. Our technical modifications will be of tangible benefit to researchers looking for a more efficient and reliable shRNA construction process.
| S-EPMC1343552 | biostudies-literature
Project description:Aerobic anoxygenic phototrophs: their role in carbon utilization under light in freshwater ecosystems
| PRJEB41596 | ENA
Project description:The strain-promoted azide-alkyne cycloaddition represents an optimal metal-free method for the modular coupling of amphiphilic polymer blocks. Hydrophilic poly(oxazoline) (PMOXA) or poly(ethylene glycol) (PEG) A-blocks were coupled with a hydrophobic poly(siloxane) B-block to provide triblock copolymers capable of self-assembling into vesicular nanostructures. Stealth properties investigated via a complement activation assay revealed the superior in vitro stealth attributes of polymeric vesicles synthesized via a metal-free approach to those coupled via the widely used copper-catalyzed click method. Furthermore, the ability to change a single parameter, such as the hydrophilic block, allowed the direct comparison of the biocompatibility properties of triblock copolymers containing PMOXA or PEG. Our studies convincingly demonstrate the need for a metal-free approach, both in preventing cytotoxicity while imparting optimal stealth properties for potential biomedical applications.
| S-EPMC3817994 | biostudies-literature
Project description:In this paper, we discuss the design study for a brain SPECT imaging system, referred to as the HelmetSPECT system, based on a spherical synthetic compound-eye (SCE) gamma camera design. The design utilizes a large number ( ∼ 500 ) of semiconductor detector modules, each coupled to an aperture with a very narrow opening for high-resolution SPECT imaging applications. In this study, we demonstrate that this novel system design could provide an excellent spatial resolution, a very high sensitivity, and a rich angular sampling without scanning motion over a clinically relevant field-of-view (FOV). These properties make the proposed HelmetSPECT system attractive for dynamic imaging of epileptic patients during seizures. In ictal SPECT, there is typically no prior information on where the seizures would happen, and both the imaging resolution and quantitative accuracy of the dynamic SPECT images would provide critical information for staging the seizures outbreak and refining the plans for subsequent surgical intervention.We report the performance evaluation and comparison among similar system geometries using non-conventional apertures, such as micro-ring and micro-slit, and traditional lofthole apertures. We demonstrate that the combination of ultrahigh-resolution imaging detectors, the SCE gamma camera design, and the micro-ring and micro-slit apertures would offer an interesting approach for the future ultrahigh-resolution clinical SPECT imaging systems without sacrificing system sensitivity and FOV.
| S-EPMC8711775 | biostudies-literature
Project description:Plant synthetic biology requires software tools to assist on the design of complex multi-genic expression plasmids. Here a vector design strategy to express genes in plants is formalized and implemented as a grammar in GenoCAD, a Computer-Aided Design software for synthetic biology. It includes a library of plant biological parts organized in structural categories and a set of rules describing how to assemble these parts into large constructs. Rules developed here are organized and divided into three main subsections according to the aim of the final construct: protein localization studies, promoter analysis and protein-protein interaction experiments. The GenoCAD plant grammar guides the user through the design while allowing users to customize vectors according to their needs. Therefore the plant grammar implemented in GenoCAD will help plant biologists take advantage of methods from synthetic biology to design expression vectors supporting their research projects.
| S-EPMC4492961 | biostudies-literature
Project description:BackgroundSalmonella enterica serotype Typhimurium is a nontyphoidal and common foodborne pathogen that causes serious threat to humans. There is no licensed vaccine to prevent the nontyphoid bacterial infection caused by S. Typhimurium.MethodsTo develop conjugate vaccines, the bacterial lipid-A free lipopolysaccharide (LFPS) is prepared as the immunogen and used to synthesize the LFPS-linker-protein conjugates 6a-9b. The designed bifunctional linkers 1-5 comprising either an o-phenylenediamine or amine moiety are specifically attached to the exposed 3-deoxy-D-manno-octulosonic acid (Kdo), an α-ketoacid saccharide of LFPS, via condensation reaction or decarboxylative amidation. In addition to bovine serum albumin and ovalbumin, the S. Typhimurium flagellin (FliC) is also used as a self-adjuvanting protein carrier.ResultsThe synthesized conjugate vaccines are characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and fast performance liquid chromatography (FPLC), and their contents of polysaccharides and protein are determined by phenol-sulfuric acid assay and bicinchoninic acid assay, respectively. Enzyme-linked immunosorbent assay (ELISA) shows that immunization of mouse with the LFPS-linker-protein vaccines at a dosage of 2.5 μg is sufficient to elicit serum immunoglobulin G (IgG) specific to S. Typhimurium lipopolysaccharide (LPS). The straight-chain amide linkers in conjugates 7a-9b do not interfere with the desired immune response. Vaccines 7a and 7b derived from either unfractionated LFPS or the high-mass portion show equal efficacy in induction of IgG antibodies. The challenge experiments are performed by oral gavage of S. Typhimurium pathogen, and vaccine 7c having FliC as the self-adjuvanting protein carrier exhibits a high vaccine efficacy of 74% with 80% mice survival rate at day 28 post the pathogen challenge.ConclusionsThis study demonstrates that lipid-A free lipopolysaccharide prepared from Gram-negative bacteria is an appropriate immunogen, in which the exposed Kdo is connected to bifunctional linkers to form conjugate vaccines. The decarboxylative amidation of Kdo is a novel and useful method to construct a relatively robust and low immunogenic straight-chain amide linkage. The vaccine efficacy is enhanced by using bacterial flagellin as the self-adjuvanting carrier protein.
| S-EPMC7443816 | biostudies-literature
Project description:Polymeric materials have been used in a range of pharmaceutical and biotechnology products for more than 40 years. These materials have evolved from their earlier use as biodegradable products such as resorbable sutures, orthopaedic implants, macroscale and microscale drug delivery systems such as microparticles and wafers used as controlled drug release depots, to multifunctional nanoparticles (NPs) capable of targeting, and controlled release of therapeutic and diagnostic agents. These newer generations of targeted and controlled release polymeric NPs are now engineered to navigate the complex in vivo environment, and incorporate functionalities for achieving target specificity, control of drug concentration and exposure kinetics at the tissue, cell, and subcellular levels. Indeed this optimization of drug pharmacology as aided by careful design of multifunctional NPs can lead to improved drug safety and efficacy, and may be complimentary to drug enhancements that are traditionally achieved by medicinal chemistry. In this regard, polymeric NPs have the potential to result in a highly differentiated new class of therapeutics, distinct from the original active drugs used in their composition, and distinct from first generation NPs that largely facilitated drug formulation. A greater flexibility in the design of drug molecules themselves may also be facilitated following their incorporation into NPs, as drug properties (solubility, metabolism, plasma binding, biodistribution, target tissue accumulation) will no longer be constrained to the same extent by drug chemical composition, but also become in-part the function of the physicochemical properties of the NP. The combination of optimally designed drugs with optimally engineered polymeric NPs opens up the possibility of improved clinical outcomes that may not be achievable with the administration of drugs in their conventional form. In this critical review, we aim to provide insights into the design and development of targeted polymeric NPs and to highlight the challenges associated with the engineering of this novel class of therapeutics, including considerations of NP design optimization, development and biophysicochemical properties. Additionally, we highlight some recent examples from the literature, which demonstrate current trends and novel concepts in both the design and utility of targeted polymeric NPs (444 references).
| S-EPMC3684255 | biostudies-other