Biochemical, Biophysical and Functional Characterization of an Insoluble Iron Containing Hepcidin–Ferritin Chimeric Monomer Assembled Together with Human Ferritin H/L Chains at Different Molar Ratios

Hepcidin and ferritin are key proteins of iron homeostasis in mammals. In this study, we characterize a chimera by fusing camel hepcidin to a human ferritin H-chain to verify if it retained the properties of the two proteins. The construct (HepcH) is expressed in E. coli in an insoluble and iron-containing form. To characterize it, the product was incubated with ascorbic acid and TCEP to reduce and solubilize the iron, which was quantified with ferrozine. HepcH bound approximately five times more iron than the wild type human ferritin, due to the presence of the hepcidin moiety. To obtain a soluble and stable product, the chimera was denatured and renatured together with different amounts of L-ferritin of the H-chain in order to produce 24-shell heteropolymers with different subunit proportions. They were analyzed by denaturing and non-denaturing PAGE and by mass spectroscopy. At the 1:5 ratio of HepcH to H- or L-ferritin, a stable and soluble molecule was obtained. Its biological activity was verified by its ability to both bind specifically cell lines that express ferroportin and to promote ferroportin degradation. This chimeric molecule showed the ability to bind both mouse J774 macrophage cells, as well as human HepG2 cells, via the hepcidin–ferroportin axis. We conclude that the chimera retains the properties of both hepcidin and ferritin and might be exploited for drug delivery.

Electrochemical oxidation of ferricyanide

We report the electrochemical oxidation of ferricyanide, [FeIII(CN)6]3− and characterised the oxidation product by in-situ FTIR and XAS spectroelectrochemistry methods. Oxidation of [FeIII(CN)6]3− is proposed to proceed via a tentative Fe(IV) intermediate that undergoes reduction elimination to give cis-[FeIII(CN)4(CH3CN)2]1− as stable product in acetonitrile. Speciation of the oxidation product by DFT calculations is underpinned by good agreement to experimental data.

Contact-Mediated Nucleation of Subcooled Droplets in Melt Emulsions: A Microfluidic Approach

The production of melt emulsions is mainly influenced by the crystallization step, as every single droplet needs to crystallize to obtain a stable product with a long shelf life. However, the crystallization of dispersed droplets requires high subcooling, resulting in a time, energy and cost intensive production processes. Contact-mediated nucleation (CMN) may be used to intensify the nucleation process, enabling crystallization at higher temperatures. It describes the successful inoculation of a subcooled liquid droplet by a crystalline particle. Surfactants are added to emulsions/suspensions for their stabilization against coalescence or aggregation. They cover the interface, lower the specific interfacial energy and form micelles in the continuous phase. It may be assumed that micelles and high concentrations of surfactant monomers in the continuous phase delay or even hinder CMN as the two reaction partners cannot get in touch. Experiments were carried out in a microfluidic chip, allowing for the controlled contact between a single subcooled liquid droplet and a single crystallized droplet. We were able to demonstrate the impact of the surfactant concentration on the CMN. Following an increase in the aqueous micelle concentrations, the time needed to inoculate the liquid droplet increased or CMN was prevented entirely.

Study on the Product Formation of the Reaction between Criegee Compound and Propargyl Radical

Mechanism of the reaction between Criegee compound (CH2OO) and Propargyl radical (C3H3) has been studied by using the density functional theory DFT/M06-2X in conjunction with the 6-311++G(3df,2p) basis set for both optimization and single-point energy calculations. The calculated results indicate that mechanism of the C3H3 + CH2OO reaction can occur in two different directions: H-atom abstraction and/or addition. As a result, 11 various products have been created from this reaction; in which, P10 (OCHCHCHCHO + H) is the most thermodynamically stable product and the reaction path leading to the P7 (CH2-[cyc-CCHCHOO] + H) product is the most energetically and kinetically favorable channel.

Formulation and Safety Tests of a Wickerhamomyces anomalus–Based Product: Potential Use of Killer Toxins of a Mosquito Symbiotic Yeast to Limit Malaria Transmission

Wickerhamomyces anomalus strain WaF17.12 is a yeast with an antiplasmodial property based on the production of a killer toxin. For its symbiotic association with Anopheles mosquitoes, it has been proposed for the control of malaria. In an applied view, we evaluated the yeast formulation by freeze-drying WaF17.12. The study was carried out by comparing yeast preparations stored at room temperature for different periods, demonstrating that lyophilization is a useful method to obtain a stable product in terms of cell growth reactivation and maintenance of the killer toxin antimicrobial activity. Moreover, cytotoxic assays on human cells were performed, showing no effects on the cell viability and the proinflammatory response. The post-formulation effectiveness of the killer toxin and the safety tests indicate that WaF17.12 is a promising bioreagent able to impair the malaria parasite in vector mosquitoes.

Fate of Functional Bacterial and Eukaryotic Community Regulated by Earthworms during Vermicomposting of Dewatered Sludge, Studies Based on the 16S rDNA and 18S rDNA Sequencing of Active Cells

DNA sequencing of active cells involved in vermicomposting can clarify the roles of earthworms in regulating functional microorganisms. This study aimed to investigate the effect of earthworms on functional microbial communities in sludge by comparing biodegradation treatments with and without earthworms. PCR and high throughput sequencing based on pretreatment of propidium monoazide (PMA) were used to detect the changes in active bacterial 16S rDNA and eukaryotic 18S rDNA during vermicomposting. The results showed that the nitrate in sludge vermicomposting and control were significantly different from day 10, with a more stable product at day 30 of vermicomposting. Compared with the control, the Shannon indexes of active bacteria and eukaryotes decreased by 1.9% and 31.1%, respectively, in sludge vermicompost. Moreover, Proteobacteria (36.2%), Actinobacteria (25.6%), and eukaryotic Cryptomycota (80.3%) were activated in the sludge vermicompost. In contrast, the control had Proteobacteria (44.8%), Bacteroidetes (14.2%), Cryptomycota (50.00%), and Arthropoda (36.59%). Network analysis showed that environmental factors had different correlations between active bacterial and eukaryotic community structures. This study suggests that earthworms can decrease the diversity of bacterial and eukaryotic communities, forming a specific-functional microbial community and thus accelerating organic matter decomposition during vermicomposting of dewatered sludge.

Techno-functional, textural and sensorial properties of frankfurters as affected by the addition of bee pollen powder

The objective of this study was to determine whether the addition of different pollen powder concentrations (0.5; 1.0 and 1.5 g/100 g) had an influence on techno-functional, textural and sensorial traits of frankfurters. Examining the techno-functional characteristics of pollen, a conclusion was reached that the higher the concentration, the higher the emulsification and better techno-functional properties. Also, FTIR-ATR analysis has shown that specific pollen molecules provided good emulsifying properties of sausages. On the other hand, sensory analysis showed that sausages with the addition of 1.0% and 1.5% of pollen powder have a more pronounced floral odor. Warner-Bratzler shear force test has shown that the incorporation of pollen caused a more stable product throughout sixty days of storage than the control sample. It could be explained by the formation of more protein-protein interactions due to the addition of non-meat proteins in the formulation of frankfurters and obtaining a more stable product than the control one. All things considered, it can be concluded that pollen exhibits good techno-functional properties and could be utilized in the formulation of frankfurters with improved and steady techno-functional properties during two months of refrigerated storage.

Development of cefdinir loaded Functionalized carbon Nanotubes dry powder Inhaler for the Treatment of cystic Fibrosis

A dry-powder inhaler (DPI) carries medication to lungs as a dry powder, useful against respiratory diseases. The current research was endeavoured to examine the capabilities of Multi-walled carbon nanotubes (MWCNT) as a pulmonary transporter for directing cefdinir to cystic fibrosis (CF). Functionalized MWCNTs were loaded with cefdinir to formulate DPI (F-CEF FMWCNTs DPI) having efficient treatment against lung infections and were evaluated successfully. The outcomes demonstrated that cefdinir loaded FMWCNTs were non-toxic and accomplished 79.73 % entrapment with better flow properties. The optimized formulation had Mass Median Aerodynamic Diameter (MMAD), Fine particle fraction (FPF), and particle size of 3.45±0.09 μm, 58.52±1.06%, 5.25 ± 0.03 μm (CEF FMWCNT DPI) and 4. 29±0.16μm 38.74±1.02%, 7.54 ± 0.02 μm (C-DPI) respectively. The loaded nanotubes showed 72. 63 % release after 15 hours in a controlled manner. The outcome of work recognized a unique, simple, and stable product having improved drug loading and increased dispersibility of carbon nanotubes (CNTs) thus improved bioavailability at a lung infection place with less adverse actions.

The Impact of Drying and Rehydration on the Structural Properties and Quality Attributes of Pre-Cooked Dried Beans

Fresh common beans can be made ‘instant’ to produce fast-cooking beans by first soaking and cooking the beans before drying to create a shelf-stable product that can be rehydrated at the time of use. This study investigated the interplay between the drying process (air, vacuum and freeze drying), the microstructure and functional attributes of rehydrated pre-cooked beans. The microscopic study revealed that the three different drying techniques resulted in distinctly different microstructures, with the freeze drying process resulting in highly porous materials, while the air- and vacuum-dried samples underwent shrinkage. Additionally, the rehydration behavior (modeled using empirical and diffusion models) demonstrates that the high rehydration rate of freeze-dried beans is due to capillarity, while rehydration, in the case of air- and vacuum-dried beans, is primarily diffusion-controlled. Irrespective of the drying technique, the high rehydration capacity supports little to no structural collapse or damage to the cell walls. The color and texture of the rehydrated beans did not differ greatly from those of freshly cooked beans. The total peak area of the volatiles of rehydrated beans was significantly reduced by the drying process, but volatiles characteristic of the cooked bean aroma were retained. This new understanding is beneficial in tailoring the functional properties of pre-cooked dry convenient beans requiring short preparation times.