Highly efficient enzyme-functionalized porous zirconia microtubes for bacteria filtration

In contrast to polymer membranes, ceramic membranes offer considerable advantages for safe drinking water provision due to their excellent chemical, thermal, and mechanical endurance. In this study, porous ceramic microtubes made of yttria stabilized zirconia (YSZ) are presented, which are conditioned for bacteria filtration by immobilizing lysozyme as an antibacterial enzyme. In accordance with determined membrane pore sizes of the nonfunctionalized microtube of ≤200 nm, log reduction values (LRV) of nearly 3 (i.e., bacterial retention of 99.9%) were obtained for bacterial retention studies using gram-positive model bacterium Micrococcus luteus. Immobilization studies of lysozyme on the membrane surface reveal an up to six times higher lysozyme loading for the covalent immobilization route as compared to unspecific immobilization. Antibacterial activity of lysozyme-functionalized microtubes was assessed by qualitative agar plate test using Micrococcus luteus as substrate showing that both the unspecific and the covalent lysozyme immobilization enhance the microtubes' antibacterial properties. Quantification of the enzyme activity at flow conditions by photometric assays reveals that the enzyme activities of lysozyme-functionalized microtubes depend strongly on applied flow rates. Intracapillary feeding of bacteria solution and higher flow rates lead to reduced enzyme activities. In consideration of different applied flow rates in the range of 0.2-0.5 mL/min, the total lysozyme activity increases by a factor of 2 for the covalent immobilization route as compared to the unspecific binding. Lysozyme leaching experiments at flow conditions for 1 h show a significant higher amount of washed-out lysozyme (factor 1.7-3.4) for the unspecific immobilization route when compared to the covalent route where the initial level of antibacterial effectiveness could be achieved by reimmobilization with lysozyme. The presented platform is highly promising for sustainable bacteria filtration.     

Direct determination of polymerised triacylglycerides in deep-frying vegetable oil by near infrared spectroscopy using Partial Least Squares regression

A green method for the determination of polymerised triacylglyceride (PTG) in deep-frying vegetable oils of different botanic origin has been developed employing near infrared (NIR) spectroscopy and Partial Least Squares (PLS) regression. Four different types of oil were heated during several hours, with and without the addition of foodstuff. NIR transmission spectra were obtained directly from sample aliquots stored in glass vials, thus avoiding the consumption of solvents and minimising waste generation. Variables employed for building the PLS models were selected applying interval PLS (iPLS) as well as Uninformative Variable Elimination-PLS (UVE-PLS). A global PLS model using spectra of all four types of oils was compared to PLS models established for each oil type. Due to the small differences observed in the NIR spectra that can be related to the different botanic origin and results obtained from the PLS model comparison, the use of a global PLS model is recommended leading to prediction errors of 2.28% (w/w) for the determination of PTG in oils employed for frying different kinds of foods.     

Analysis of Phenolic Composition of Noble Muscadine (Vitis rotundifolia) by HPLC‐MS and the Relationship to Its Antioxidant Capacity

Abstract: Phenolic compounds and anthocyanins in muscadines have attracted much attention due to their diverse biological activities. With bioassays of antioxidant activities in terms of total phenolic content (TPC), total anthocyanin content (TAC), total procyanidin content (TPA), oxygen radical absorbance capacity (ORAC), and ferric reducing antioxidant power (FRAP) of different parts of the Noble muscadine, the butanol (BuOH) extract of the muscadine skin showed the highest TPC (317.91 ± 1.83 mg GAE/100 g FW), which might be ascribed to its high TAC of 227.06 ± 1.29 mg/100 g fresh weight (FW). The ethyl acetate (EtOAc) extract of the muscadine seed contained the highest TPA (55.30 ± 0.63 mg CE/100 g FW). Correlation analyses demonstrated a significant linear relationship of TPC and TAC compared to their ORAC and FRAP values within the range of R² from 0.9283 to 0.9936, which suggested that phenolics and anthocyanins in the extracts contributed significantly to their antioxidant potential. Nineteen individual phenolics and 5 anthocyanins were identified by HPLC‐MS, which indicated different chemical profiles of anthocyanins and other phenolics in the muscadine extracts. Practical Application: The paper has provided rich information of bioactive phytochemical profiles in different solvent extracts and their correlation with the antioxidant activity in the muscadine that is a very special regional fruit in U.S. Its high content of phenolic compounds demonstrates that muscadine could be beneficial to human health.     

Structural changes,chemical composition and antioxidant activity of cherry tomato fruits (cv. Micro‐Tom) stored under optimal and chilling conditions

BACKGROUND: Cherry tomato fruits cv. Micro‐Tom, a model plant for tomato genetics, were analysed in order to determine the main structural and chemical changes under optimal and chilling storage conditions. The comparison of Micro‐Tom to standard tomato cultivars will give an insight into suitability of this dwarf cultivar as a model for studying the influence of low‐temperature conditions on tomato fruits. RESULTS: During chilling, fruit tissue was progressively destroyed due to the collapse of the deep layers of the pericarp. Chilling lowered the typical tomato kinetics of ripening in sugars (glucose, fructose, sucrose), organic acids (tartaric, malic, citric, ascorbic and succinic) and the antioxidants phenol and lycopene, while carotenoid synthesis seemed to be blocked. Glutathione level was elevated and the activity of ROS scavenging enzymes was altered. Essentially, Micro‐Tom fruits showed a quality evolution that was similar to that described for standard tomato cultivars. CONCLUSION: The cherry tomato cultivar Micro‐Tom could be used as a model for studying the influence of low temperature on biochemical and structural changes taking place during chilling injury conditions. Copyright © 2009 Society of Chemical Industry     

Simulations of microfluidic droplet formation using the two-phase level set method

Microdroplet formation is an emerging area of research due to its wide-ranging applications within microfluidic based lab-on-a-chip devices. Our goal is to understand the dynamics of droplet formation in a microfluidic T-junction in order to optimize the operation of the microfluidic device. Understanding of this process forms the basis of many potential applications: synthesis of new materials, formulation of products in pharmaceutical, cosmetics and food industries. The two-phase level set method, which is ideally suited for tracking the interfaces between two immiscible fluids, has been used to perform numerical simulations of droplet formation in a T-junction. Numerical predictions compare well with experimental observations. The influence of parameters such as flow rate ratio, capillary number, viscosity ratio and the interfacial tension between the two immiscible fluids is known to affect the physical processes of droplet generation. In this study the effects of surface wettability, which can be controlled by altering the contact angle, are investigated systematically. As competitive wetting between liquids in a two-phase flow can give rise to erratic flow patterns, it is often desirable to minimize this phenomenon as it can lead to a disruption of the regular production of uniform droplets. The numerical simulations predicted that wettability effects on droplet length are more prominent when the viscosity ratio λ (the quotient of the viscosity of the dispersed phase with the viscosity of the continuous phase) is O(1), compared to the situation when λ is O(0.1). The droplet size becomes independent of contact angle in the superhydrophobic regime for all capillary numbers. At a given value of interfacial tension, the droplet length is greater when λ is O(1) compared to the case when λ is O(0.1). The increase in droplet length with interfacial tension, σ, is a function of with the coefficients of the regression curves depending on the viscosity ratio.     

Primary Cilia Structure Is Prolonged in Enteric Neurons of 5xFAD Alzheimer’s Disease Model Mice

Neurodegenerative diseases such as Alzheimer’s disease (AD) have long been acknowledged as mere disorders of the central nervous system (CNS). However, in recent years the gut with its autonomous nervous system and the multitude of microbial commensals has come into focus. Changes in gut properties have been described in patients and animal disease models such as altered enzyme secretion or architecture of the enteric nervous system. The underlying cellular mechanisms have so far only been poorly investigated. An important organelle for integrating potentially toxic signals such as the AD characteristic A-beta peptide is the primary cilium. This microtubule-based signaling organelle regulates numerous cellular processes. Even though the role of primary cilia in a variety of developmental and disease processes has recently been recognized, the contribution of defective ciliary signaling to neurodegenerative diseases such as AD, however, has not been investigated in detail so far. The AD mouse model 5xFAD was used to analyze possible changes in gut functionality by organ bath measurement of peristalsis movement. Subsequently, we cultured primary enteric neurons from mutant mice and wild type littermate controls and assessed for cellular pathomechanisms. Neurite mass was quantified within transwell culturing experiments. Using a combination of different markers for the primary cilium, cilia number and length were determined using fluorescence microscopy. 5xFAD mice showed altered gut anatomy, motility, and neurite mass of enteric neurons. Moreover, primary cilia could be demonstrated on the surface of enteric neurons and exhibited an elongated phenotype in 5xFAD mice. In parallel, we observed reduced β-Catenin expression, a key signaling molecule that regulates Wnt signaling, which is regulated in part via ciliary associated mechanisms. Both results could be recapitulated via in vitro treatments of enteric neurons from wild type mice with A-beta. So far, only a few reports on the probable role of primary cilia in AD can be found. Here, we reveal for the first time an architectural altered phenotype of primary cilia in the enteric nervous system of AD model mice, elicited potentially by neurotoxic A-beta. Potential changes on the sub-organelle level—also in CNS-derived neurons—require further investigations.     

Sequence Features of Drosha and Dicer Cleavage Sites Affect the Complexity of IsomiRs

The deep-sequencing of small RNAs has revealed that different numbers and proportions of miRNA variants called isomiRs are formed from single miRNA genes and that this effect is attributable mainly to imprecise cleavage by Drosha and Dicer. Factors that influence the degree of cleavage precision of Drosha and Dicer are under investigation, and their identification may improve our understanding of the mechanisms by which cells modulate the regulatory potential of miRNAs. In this study, we focused on the sequences and structural determinants of Drosha and Dicer cleavage sites, which may explain the generation of homogeneous miRNAs (in which a single isomiR strongly predominates) as well as the generation of heterogeneous miRNAs. Using deep-sequencing data for small RNAs, we demonstrate that the generation of homogeneous miRNAs requires more sequence constraints at the cleavage sites than the formation of heterogeneous miRNAs. Additionally, our results indicate that specific Drosha cleavage sites have more sequence determinants in miRNA precursors than specific cleavage sites for Dicer and that secondary structural motifs in the miRNA precursors influence the precision of Dicer cleavage. Together, we present the sequence and structural features of Drosha and Dicer cleavage sites that influence the heterogeneity of the released miRNAs.     

From ELF to Compressibility in Solids

Understanding the electronic nature of materials’ compressibility has always been a major issue behind tabulation and rationalization of bulk moduli. This is especially because this understanding is one of the main approaches to the design and proposal of new materials with a desired (e.g., ultralow) compressibility. It is well recognized that the softest part of the solid will be the one responsible for its compression at the first place. In chemical terms, this means that the valence will suffer the main consequences of pressurization. It is desirable to understand this response to pressure in terms of the valence properties (charge, volume, etc.). One of the possible approaches is to consider models of electronic separability, such as the bond charge model (BCM), which provides insight into the cohesion of covalent crystals in analogy with the classical ionic model. However, this model relies on empirical parametrization of bond and lone pair properties. In this contribution, we have coupled electron localization function (ELF) ab initio data with the bond charge model developed by Parr in order to analyze solid state compressibility from first principles and moreover, to derive general trends and shed light upon superhard behavior.     

Quantitative Analysis of PMLA Nanoconjugate Components after Backbone Cleavage

Multifunctional polymer nanoconjugates containing multiple components show great promise in cancer therapy, but in most cases complete analysis of each component is difficult. Polymalic acid (PMLA) based nanoconjugates have demonstrated successful brain and breast cancer treatment. They consist of multiple components including targeting antibodies, Morpholino antisense oligonucleotides (AONs), and endosome escape moieties. The component analysis of PMLA nanoconjugates is extremely difficult using conventional spectrometry and HPLC method. Taking advantage of the nature of polyester of PMLA, which can be cleaved by ammonium hydroxide, we describe a method to analyze the content of antibody and AON within nanoconjugates simultaneously using SEC-HPLC by selectively cleaving the PMLA backbone. The selected cleavage conditions only degrade PMLA without affecting the integrity and biological activity of the antibody. Although the amount of antibody could also be determined using the bicinchoninic acid (BCA) method, our selective cleavage method gives more reliable results and is more powerful. Our approach provides a new direction for the component analysis of polymer nanoconjugates and nanoparticles.