Enzyme-functionalized hydrogel film for extracorporeal uric acid reduction.

Enzyme replacement therapy for hyperuricemia treatment has been proven effective for critical state hyperuricemia patients. Still, direct administration of recombinant uricase can induce several fatal side effects. To circumvent this drawback, hydrogel protein carriers can be used in platforms for extracorporeal treatment such as microscale-based devices. In this work, calcium alginate and poly-(vinyl alcohol) hydrogel films were studied for their urate oxidase immobilization and uric acid reduction, which could be implemented in microscale-based extracorporeal devices. A mathematical model was developed in conjunction with uric acid reduction experiments to evaluate the influence of mass transfer and reaction parameters in the Michaelis-Menten kinetic expression. Alginate hydrogels prepared with crosslinker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-(hydroxysuccinimide) offered superior diffusivity of uric acid in the gel matrix at the maximum value of D g , UA ≈ $$ {D}_{\mathrm{g},\mathrm{UA}}\approx $$ 1.98 × 10-11 m2 /s compared with alginate prepared solely from ionic crosslinking with D g , UA ≈ $$ {D}_{\mathrm{g},\mathrm{UA}}\approx $$ 5.31 × 10-12 m2 /s at the same alginate concentration. The maximum value of νmax was experimentally determined at 7.78 × 10-5 mol/(m3 s). A 3% sodium alginate hydrogel with crosslinkers yielded the highest reduction of uric acid at 92.70%. The mathematical model demonstrated an excellent prediction of uric acid conversion suggesting potential use of the model for formulation and maximizing the therapeutic performance of functionalized hydrogels.

Correlation Theory of the Maxillary Central Incisor, Face and Dental Arch Shape in the Serbian Population.

Background and Objectives: According to the modified Williams theory, the shape of the maxillary central incisor corresponds to the shape of the maxillary alveolar ridge (dental arch) and the shape of the face. Moreover, the standards of beauty suggest that the center of the face of an individual with a full set of teeth should match the center of the maxillary and mandibular dental arches. The purpose of this study is to conduct a comparative cross-sectional study on the matching of the shape of the face, maxillary central incisor and maxillary dental arch as well as the matching of the midfacial line and dental arches in subjects with complete dentition. Materials and Methods: The study included 90 subjects of both sexes. The matching of the shape of the face and midfacial line, i.e., dental arches and maxillary incisors, was determined by analysing photographs, whereas the shape of the dental arch was determined by analysing plaster models. Results: No significant gender-related differences were found either in the shape of the maxillary central incisor that matched the shape of the maxillary dental arch (p = 0.349) or in the shape of the dental arch that matched the shape of the face (p = 0.697). However, a significant difference was noted in the shape of the teeth that matched the shape of the face (p = 0.043), which was more significantly impaired in men. In addition, the matching of the mid-face and the mid-dental arch was significantly greater in women (p = 0.016). Conclusions: The modified Williams theory was confirmed in most subjects, thus it can be considered a relevant guideline when determining the shape of teeth after their loss. The highest percentage of matching in both sexes was with the shape of the face and dental arch. There was no positive correlation between the middle of the maxillary and mandibular dental arch in most cases.

Longevity Demonstration of Methane to C2 via a Nonthermal Plasma Microreactor.

Hydrocarbon processing using plasmas has tremendous potential, yet there still exist many uncertainties pertaining to practical operation over long durations. Previously, it has been demonstrated that a nonthermal plasma operating in a DC glow regime can transform methane into C2 species (acetylene, ethylene, ethane) in a microreactor. Using a DC glow regime in a microchannel reactor allows for lower power consumption, at the expense of greater consequence of fouling. Since biogas can be a source of methane, a longevity study was undertaken to understand how the microreactor system would change over time with a feed mixture of simulated biogas (CO2, CH4) and air. Two different biogas mixtures were used, one of which contained 300 ppm H2S, while the other had no H2S. Potential difficulties observed from previous experiments included carbon deposition on the electrodes, which could interfere with the electrical characteristics of the plasma discharge as well as material deposition in the microchannel, which could affect gas flow. It was found that raising the temperature of the system to 120 °C helped prevent hydrocarbon deposition in the reactor. Purging the reactor periodically with dry air was also found to have positive effects as it removed carbon buildup on the electrodes themselves. Successful operation over a 50 h time period without any significant deterioration was demonstrated.

Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants.

Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25-43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1-24, the truncation at position 31 is predicted to change the structure within aa 15-31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.

Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides.

AIMS: The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. METHODS AND RESULTS: To improve the pMAL expression vector, we introduced the His6 tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His6 -Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His6 tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His6 and His6 -enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human ß-defensin. CONCLUSIONS: We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. SIGNIFICANCE AND IMPACT OF THE STUDY: Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.

The route to transcription initiation determines the mode of transcriptional bursting in E. coli.

Transcription is fundamentally noisy, leading to significant heterogeneity across bacterial populations. Noise is often attributed to burstiness, but the underlying mechanisms and their dependence on the mode of promotor regulation remain unclear. Here, we measure E. coli single cell mRNA levels for two stress responses that depend on bacterial sigma factors with different mode of transcription initiation (σ70 and σ54). By fitting a stochastic model to the observed mRNA distributions, we show that the transition from low to high expression of the σ70-controlled stress response is regulated via the burst size, while that of the σ54-controlled stress response is regulated via the burst frequency. Therefore, transcription initiation involving σ54 differs from other bacterial systems, and yields bursting kinetics characteristic of eukaryotic systems.

Comparative antiradical activity and molecular Docking/Dynamics analysis of octopamine and norepinephrine: the role of OH groups.

Octopamine is a neurotransmitter in invertebrates and a phenol analog of norepinephrine. The crystallographic and spectral (UV-visUV, and NMR) characteristics of octopamine were investigated experimentally and theoretically by applying appropriate level of theory, B3LYP-D3BJ/6-311++G(d,p), which reproduced well the experimental bond lengths and angles. The intramolecular interactions governing the stability of conformers were described by NBO and QTAIM analyses. The antiradical potencies of octopamine and norepinephrine towards DPPH and ABTS+ were examined with special emphasis on the preferred mechanism and effect of catechol moiety. Several techniques were used to distinguish Hydrogen Atom Transfer (HAT) and Proton Coupled Electron Transfer (PCET) mechanisms for reaction with DPPH. The calculated rate constants of the reactions with both radicals showed that Sequential Proton Loss Electron Transfer (SPLET) mechanism was dominant both thermodynamically and kinetically, with values of thermodynamic functions and rate constants clearly proving the importance of the second hydroxyl group in structure. The Molecular Docking and afterward Molecular Dynamics calculations of formed complexes between octopamine/norepinephrine with ß1- and ß2- adrenergic receptors examined in details the interactions that lead to the formation of stable complexes. The number of strong interactions of amino acids with norepinephrine was higher, but the absence of hydroxyl group in octopamine did not lead to a significant change in the type of interactions and stability. The formed complexes showed higher flexibility of amino acids, similar compactness of structure as proteins and increased interatomic distances of the backbone when compared to pure proteins.

Effects of coenzyme Q10 encapsulated in nanoliposomes on wound healing processes after tooth extraction.

BACKGROUND/PURPOSE: Tooth extraction is often followed by a number of different complications that demand additional treatment. In order to accelerate healing processes and decrease the complication occurrence various agents, growth factors, natural and synthetic antioxidants (e.g coenzyme Q10-CoQ10), are applied. Due to the partially known health-promoting effects of CoQ10 we decided to assess potential of it's encapsulated in nanoliposomes form on wound healing process following tooth extraction. MATERIALS AND METHODS: Effects of free and encapsulated form of CoQ10 on wound healing processes after tooth extraction in rats, 3 and 7 days following surgical procedure, was studied by means of tissue biochemical (myeloperoxidase activity and nitric oxide (NO) concentrations) and pathohistological analysis. RESULTS: The obtained results indicate that the encapsulated form of CoQ10 compared to control and CoQ10 treated animals statistically significantly decreases inflammatory process estimated through myeloperoxidase activity and NO concentrations, as well as based on histopathological analysis 3 and 7 days following surgery. CONCLUSION: The results of this study unequivocally prove that the encapsulation of CoQ10 in nanoliposomes enhances CoQ10 activity by accelerating wound healing process after tooth extraction.

Structure and function of PspA and Vipp1 N-terminal peptides: Insights into the membrane stress sensing and mitigation.

The phage shock protein (Psp) response maintains integrity of the inner membrane (IM) in response to extracytoplasmic stress conditions and is widely distributed amongst enterobacteria. Its central component PspA, a member of the IM30 peripheral membrane protein family, acts as a major effector of the system through its direct association with the IM. Under non-stress conditions PspA also negatively regulates its own expression via direct interaction with the AAA+ ATPase PspF. PspA has a counterpart in cyanobacteria called Vipp1, which is implicated in protection of the thylakoid membranes. PspA's and Vipp1's conserved N-terminal regions contain a putative amphipathic helix a (AHa) required for membrane binding. An adjacent amphipathic helix b (AHb) in PspA is required for imposing negative control upon PspF. Here, purified peptides derived from the putative AH regions of PspA and Vipp1 were used to directly probe their effector and regulatory functions. We observed direct membrane-binding of AHa derived peptides and an accompanying change in secondary structure from unstructured to alpha-helical establishing them as bona fide membrane-sensing AH's. The peptide-binding specificities and their effects on membrane stability depend on membrane anionic lipid content and stored curvature elastic stress, in agreement with full length PspA and Vipp1 protein functionalities. AHb of PspA inhibited the ATPase activity of PspF demonstrating its direct regulatory role. These findings provide new insight into the membrane binding and function of PspA and Vipp1 and establish that synthetic peptides can be used to probe the structure-function of the IM30 protein family.

A data comparison between a traditional and the single-step ß-galactosidase assay.

This article describes reproducibility of a single-step automated ß-galactosidase, and the equivalence of its data to the traditional assay ("Experiments in Molecular Genetics" [1]). This was done via a pairwise comparison of both methods using strains with Miller Unit [MU] values ranging from 0 to over 2000. The data presented in this article is associated with the research article entitled "A single-step method for mid to high throughput ß-galactosidase assays in Escherichia coli using a microplate reader" [2].

Transcription Regulation and Membrane Stress Management in Enterobacterial Pathogens.

Transcription regulation in a temporal and conditional manner underpins the lifecycle of enterobacterial pathogens. Upon exposure to a wide array of environmental cues, these pathogens modulate their gene expression via the RNA polymerase and associated sigma factors. Different sigma factors, either involved in general 'house-keeping' or specific responses, guide the RNA polymerase to their cognate promoter DNAs. The major alternative sigma54 factor when activated helps pathogens manage stresses and proliferate in their ecological niches. In this chapter, we review the function and regulation of the sigma54-dependent Phage shock protein (Psp) system-a major stress response when Gram-negative pathogens encounter damages to their inner membranes. We discuss the recent development on mechanisms of gene regulation, signal transduction and stress mitigation in light of different biophysical and biochemical approaches.

Single-step method for ß-galactosidase assays in Escherichia coli using a 96-well microplate reader.

Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ß-d-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%.

Effect of PEO coating on bubble behavior within a polycarbonate microchannel array: A model for hemodialysis.

Obstruction of fluid flow by stationary bubbles in a microchannel hemodialyzer decreases filtration performance and increases damage to blood cells through flow maldistribution. A polyethylene oxide (PEO)-polybutadiene (PB)-polyethylene oxide surface modification, previously shown to reduce protein fouling and water/air contact angle in polycarbonate microchannel hemodialyzers, can improve microchannel wettability and may reduce bubble stagnation by lessening the resistive forces that compete with fluid flow. In this study, the effect of the PEO-PB-PEO coating on bubble retention in a microchannel array was investigated. Polycarbonate microchannel surfaces were coated with PEO-PB-PEO triblock polymer via radiolytic grafting. Channel obstruction was measured for coated and uncoated microchannels after injecting a short stream of air bubbles into the device under average nominal water velocities of 0.9 to 7.2 cm/s in the channels. The presence of the PEO coating reduced obstruction of microchannels by stationary bubbles within the range of 1.8 to 3.6 cm/s, average nominal velocity. Numerical simulations based on the lattice Boltzmann method indicate that beneficial effects may be due to the maintenance of a lubricating, thin liquid film around the bubble. The determined effective range of the PEO coating for bubble management serves as an important design constraint. These findings serve to validate the multiutility of the PEO-PB-PEO coating (bubble lubrication, biocompatibility, and therapeutic loading). © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 941-948, 2016.

Membrane Stored Curvature Elastic Stress Modulates Recruitment of Maintenance Proteins PspA and Vipp1.

UNLABELLED: Phage shock protein A (PspA), which is responsible for maintaining inner membrane integrity under stress in enterobacteria, and vesicle-inducting protein in plastids 1 (Vipp1), which functions for membrane maintenance and thylakoid biogenesis in cyanobacteria and plants, are similar peripheral membrane-binding proteins. Their homologous N-terminal amphipathic helices are required for membrane binding; however, the membrane features recognized and required for expressing their functionalities have remained largely uncharacterized. Rigorously controlled, in vitro methodologies with lipid vesicles and purified proteins were used in this study and provided the first biochemical and biophysical characterizations of membrane binding by PspA and Vipp1. Both proteins are found to sense stored curvature elastic (SCE) stress and anionic lipids within the membrane. PspA has an enhanced sensitivity for SCE stress and a higher affinity for the membrane than Vipp1. These variations in binding may be crucial for some of the proteins' differing roles in vivo. Assays probing the transcriptional regulatory function of PspA in the presence of vesicles showed that a relief of transcription inhibition occurs in an SCE stress-specific manner. This in vitro recapitulation of membrane stress-dependent transcription control suggests that the Psp response may be mounted in vivo when a cell's inner membrane experiences increased SCE stress. IMPORTANCE: All cell types maintain the integrity of their membrane systems. One widely distributed membrane stress response system in bacteria is the phage shock protein (Psp) system. The central component, peripheral membrane protein PspA, which mitigates inner membrane stress in bacteria, has a counterpart, Vipp1, which functions for membrane maintenance and thylakoid biogenesis in plants and photosynthetic bacteria. Membrane association of both these proteins is accepted as playing a pivotal role in their functions. Here we show that direct membrane binding by PspA and Vipp1 is driven by two physio-chemical signals, one of which is membrane stress specific. Our work points to alleviation of membrane stored curvature elastic stress by amphipathic helix insertions as an attractive mechanism for membrane maintenance by PspA and Vipp1. Furthermore, the identification of a physical, stress-related membrane signal suggests a unilateral mechanism that promotes both binding of PspA and induction of the Psp response.

Is the cellular and molecular machinery docile in the stationary phase of Escherichia coli?

The bacterial cell envelope retains a highly dense cytoplasm. The properties of the cytoplasm change with the metabolic state of the cell, the logarithmic phase (log) being highly active and the stationary phase metabolically much slower. Under the differing growth phases, many different types of stress mechanisms are activated in order to maintain cellular integrity. One such response in enterobacteria is the phage shock protein (Psp) response that enables adaptation to the inner membrane (IM) stress. The Psp system consists of a transcriptional activator PspF, negative regulator PspA, signal sensors PspBC, with PspA and PspG acting as effectors. The single molecule imaging of the PspF showed the existence of dynamic communication between the nucleoid-bound states of PspF and membrane via negative regulator PspA and PspBC sensors. The movement of proteins in the cytoplasm of bacterial cells is often by passive diffusion. It is plausible that the dynamics of the biomolecules differs with the state of the cytoplasm depending on the growth phase. Therefore, the Psp response proteins might encounter the densely packed glass-like properties of the cytoplasm in the stationary phase, which can influence their cellular dynamics and function. By comparing the properties of the log and stationary phases, we find that the dynamics of PspF are influenced by the growth phase and may be controlled by the changes in the cytoplasmic fluidity.

Phosphorelay of non-orthodox two component systems functions through a bi-molecular mechanism in vivo: the case of ArcB.

Two-component systems play a central part in bacterial signal transduction. Phosphorelay mechanisms have been linked to more robust and ultra-sensitive signalling dynamics. The molecular machinery that facilitates such a signalling is, however, only understood in outline. In particular the functional relevance of the dimerization of a non-orthodox or hybrid histidine kinase along which the phosphorelay takes place has been a subject of debate. We use a combination of molecular and genetic approaches, coupled to mathematical and statistical modelling, to demonstrate that the different possible intra- and inter-molecular mechanisms of phosphotransfer are formally non-identifiable in Escherichia coli expressing the ArcB non-orthodox histidine kinase used in anoxic redox control. In order to resolve this issue we further analyse the mathematical model in order to identify discriminatory experiments, which are then performed to address cis- and trans-phosphorelay mechanisms. The results suggest that exclusive cis- and trans-mechanisms will not be operating, instead the functional phosphorelay is likely to build around a sequence of allosteric interactions among the domain pairs in the histidine kinase. This is the first detailed mechanistic analysis of the molecular processes involved in non-orthodox two-component signalling and our results suggest strongly that dimerization facilitates more discriminatory proof-reading of external signals, via these allosteric reactions, prior to them being further processed.

In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription.

Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ(54)), in a purified system. We include the complete transcription system, binding interactions between σ54 and its activators, as well as the self-assembly and the critical ATPase activity of the cognate activators which serve to remodel the closed promoter complexes. We also present in vivo methodologies that are used to study the impact of physiological processes, metabolic states, global signalling networks, and cellular architecture on the control of σ(54)-dependent gene expression.

Anionic lipids and the cytoskeletal proteins MreB and RodZ define the spatio-temporal distribution and function of membrane stress controller PspA in Escherichia coli.

All cell types must maintain the integrity of their membranes. The conserved bacterial membrane-associated protein PspA is a major effector acting upon extracytoplasmic stress and is implicated in protection of the inner membrane of pathogens, formation of biofilms and multi-drug-resistant persister cells. PspA and its homologues in Gram-positive bacteria and archaea protect the cell envelope whilst also supporting thylakoid biogenesis in cyanobacteria and higher plants. In enterobacteria, PspA is a dual function protein negatively regulating the Psp system in the absence of stress and acting as an effector of membrane integrity upon stress. We show that in Escherichia coli the low-order oligomeric PspA regulatory complex associates with cardiolipin-rich, curved polar inner membrane regions. There, cardiolipin and the flotillin 1 homologue YqiK support the PspBC sensors in transducing a membrane stress signal to the PspA-PspF inhibitory complex. After stress perception, PspA high-order oligomeric effector complexes initially assemble in polar membrane regions. Subsequently, the discrete spatial distribution and dynamics of PspA effector(s) in lateral membrane regions depend on the actin homologue MreB and the peptidoglycan machinery protein RodZ. The consequences of loss of cytoplasmic membrane anionic lipids, MreB, RodZ and/or YqiK suggest that the mode of action of the PspA effector is closely associated with cell envelope organization.

Removal of the phage-shock protein PspB causes reduction of virulence in Salmonella enterica serovar Typhimurium independently of NRAMP1.

The phage-shock protein (Psp) system is believed to manage membrane stress in all Enterobacteriaceae and has recently emerged as being important for virulence in several pathogenic species of this phylum. The core of the Psp system consists of the pspA-D operon and the distantly located pspG gene. In Salmonella enterica serovar Typhimurium (S. Typhimurium), it has recently been reported that PspA is essential for systemic infection of mice, but only in NRAMP1(+) mice, signifying that attenuation is related to coping with divalent cation starvation in the intracellular environment. In the present study, we investigated the contribution of individual psp genes to virulence of S. Typhimurium. Interestingly, deletion of the whole pspA-D set of genes caused attenuation in both NRAMP1(+) and NRAMP1(-) mice, indicating that one or more of the psp genes contribute to virulence independently of NRAMP1 expression in the host. Investigations of single gene mutants showed that knock out of pspB reduced virulence in both types of mice, while deletion of pspA only caused attenuation in NRAMP1(+) mice, and deletion of pspD had a minor effect in NRAMP1(-) mice, while deletions of either pspC or pspG did not affect virulence. Experiments addressed at elucidating the role of PspB in virulence revealed that PspB is dispensable for uptake to and intracellular replication in cultured macrophages and resistance to complement-induced killing. Furthermore, the Psp system of S. Typhimurium was dispensable during pIV-induced secretin stress. In conclusion, our results demonstrate that removal of PspB reduces virulence in S. Typhimurium independently of host NRAMP1 expression, demonstrating that PspB has roles in intra-host survival distinct from the reported contributions of PspA.

Preparation and evaluation of PEO-coated materials for a microchannel hemodialyzer.

The marked increase in surface-to-volume ratio associated with microscale devices for hemodialysis leads to problems with hemocompatibility and blood flow distribution that are more challenging to manage than those encountered at the conventional scale. In this work stable surface modifications with pendant polyethylene oxide (PEO) chains were produced on polydimethylsiloxane (PDMS), polycarbonate microchannel, and polyacrylonitrile membrane materials used in construction of microchannel hemodialyzer test articles. PEO layers were prepared by radiolytic grafting of PEO-polybutadiene-PEO (PEO-PB-PEO) triblock polymers to the material surfaces. Protein repulsion was evaluated by measurement of surface-bound enzyme activity following contact of uncoated and PEO-coated surfaces with ß-galactosidase. Protein adsorption was decreased on PEO-coated polycarbonate and PDMS materials to about 20% of the level recorded on the uncoated materials. Neither the triblocks nor the irradiation process was observed to have any effect on protein interaction with the polyacrylonitrile membrane, or its permeability to urea. This approach holds promise as a means for in situ application of safe, efficacious coatings to microfluidic devices for blood processing that will ensure good hemocompatibility and blood flow distribution, with no adverse effects on mass transfer.