A novel ACE2-Based electrochemical biosensor for sensitive detection of SARS-CoV-2.

SARS-CoV-2 emerged in late 2019 and quickly spread globally, resulting in significant morbidity, mortality, and socio-economic disruptions. As of now, collaborative global efforts in vaccination and the advent of novel diagnostic tools have considerably curbed the spread and impact of the virus in many regions. Despite this progress, the demand remains for low-cost, accurate, rapid and scalable diagnostic tools to reduce the influence of SARS-CoV-2. Herein, the angiotensin-converting enzyme 2 (ACE2), a receptor for SARS-CoV-2, was immobilized on two types of electrodes, a screen-printed gold electrode (SPGE) and a screen-printed carbon electrode (SPCE), to develop electrochemical biosensors for detecting SARS-CoV-2 with high sensitivity and selectivity. This was achieved by using 1H, 1H, 2H, 2H-perfluorodecanethiol (PFDT) and aryl diazonium salt serving as linkers for SPGEs and SPCEs, respectively. Once SARS-CoV-2 was anchored onto the ACE2, the interaction of the virus with the redox probe was analyzed using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Aryl diazonium salt was observed as a superior linker compared to PFDT due to its consistent performance in the modification of the SPCEs and effective ACE2 enzyme immobilization. A distinct pair of redox peaks in the cyclic voltammogram of the biosensor modified with aryl diazonium salt highlighted the redox reaction between the functional groups of SARS-CoV-2 and the redox probe. The sensor presented a linear relationship between the redox response and the logarithm of SARS-CoV-2 concentration, with a detection limit of 1.02 × 106 TCID50/mL (50% tissue culture infectious dose). Furthermore, the biosensor showed remarkable selectivity towards SARS-CoV-2 over H1N1virus.

Effects of flow-induced electromagnetic field and surface roughness on antifouling activity of phenolic compounds.

Microbial fouling involves the physicochemical interactions between microorganisms and solid surfaces. An electromagnetic field (EMF) may change the diffusion rates of microbial cells and the electrical double layer around the cells and contacting surfaces. In the current study, polycardanol exhibiting antibiofouling activity was modified with ferromagnetic iron oxide (IO) to investigate the EMF effects on bacterial adhesion. When there was a flow of electrolyte that contained bacterial cells, flow-induced EMF was generated according to Faraday's principle. It was observed that the IO-ionic solution (IS)-modified surfaces, with an induced current of 44, 53, 66 nA, showed decreases in the adhesion of bacteria cells more than the unmodified (polycardanol) and IO-nanoparticles-modified ones. In addition to the EMF effects, the nano-scale uniform roughness of the modified surfaces appeared to play an important role in the reduction of cell adhesion. The results demonstrated that the IOIS-modified surface (3.2 × 10-6 mM IO) had the highest antibiofouling activity.

Adsorption Kinetics of Glycated Hemoglobin on Aptamer Microarrays with Antifouling Surface Modification.

Aptamers are oligonucleotides that bind with high affinity to target molecules of interest. One such target is glycated hemoglobin (gHb), a biomarker for assessing glycemic control and diabetes diagnosis. By the coupling of aptamers with surface plasmon resonance (SPR) sensing surfaces, a fast, reliable and inexpensive assay for gHb can be developed. In this study, we tested the affinity of SPR-sensing surfaces, composed of aptamers and antifouling self-assembled monolayers (SAMs), to hemoglobin (Hb) and gHb. First, we developed a gHb-targeted aptamer (GHA) through a modified Systematic Evolution of Ligands by EXponential (SELEX) enrichment process and tested its affinity to gHb using the Nano-Affi protocol. GHA was used to produce three distinct SAM-SPR-sensing surfaces: (Type-1) a SAM of GHA directly attached to a sensor surface; (Type-2) GHA attached to a SAM of 11-mercaptoundecanoic acid (11MUA) on a sensor surface; (Type-3) GHA attached to a binary SAM of 11MUA and 3,6-dioxa-8-mercaptooctan-1-ol (DMOL) on a sensor surface. Type-2 and Type-3 surfaces were characterized by cyclic voltammetry and electrochemical impedance spectroscopy to confirm that GHA bound to the underlying SAMs. The adsorption kinetics for Hb and gHb interacting with each SPR sensing surface were used to quantify their respective affinities. The Type-1 surface without antifouling modification had a dissociation constant ratio (KD,Hb/KD,gHb) of 9.7, as compared to 809.3 for the Type-3 surface, demonstrating a higher association of GHA to gHb for sensor surfaces with antifouling modifications than those without. The enhanced selectivity of GHA to gHb can likely be attributed to the inclusion of DMOL in the SAM-modified surface, which reduced interference from nonspecific adsorption of proteins. Results suggest that pairing aptamers with antifouling SAMs can significantly improve their target affinity, potentially allowing for the development of novel, low cost, and fast assays.

Mini-GAGR, an intranasally applied polysaccharide, activates the neuronal Nrf2-mediated antioxidant defense system.

Oxidative stress triggers and exacerbates neurodegeneration in Alzheimer's disease (AD). Various antioxidants reduce oxidative stress, but these agents have little efficacy due to poor blood-brain barrier (BBB) permeability. Additionally, single-modal antioxidants are easily overwhelmed by global oxidative stress. Activating nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2) and its downstream antioxidant system are considered very effective for reducing global oxidative stress. Thus far, only a few BBB-permeable agents activate the Nrf2-dependent antioxidant system. Here, we discovered a BBB-bypassing Nrf2-activating polysaccharide that may attenuate AD pathogenesis. Mini-GAGR, a 0.7-kDa cleavage product of low-acyl gellan gum, increased the levels and activities of Nrf2-dependent antioxidant enzymes, decreased reactive oxygen species (ROS) under oxidative stress in mouse cortical neurons, and robustly protected mitochondria from oxidative insults. Moreover, mini-GAGR increased the nuclear localization and transcriptional activity of Nrf2 similarly to known Nrf2 activators. Mechanistically, mini-GAGR increased the dissociation of Nrf2 from its inhibitor, Kelch-like ECH-associated protein 1 (Keap1), and induced phosphorylation and nuclear translocation of Nrf2 in a protein kinase C (PKC)- and fibroblast growth factor receptor (FGFR1)-dependent manner. Finally, 20-day intranasal treatment of 3xTg-AD mice with 100 nmol of mini-GAGR increased nuclear p-Nrf2 and growth-associated protein 43 (GAP43) levels in hippocampal neurons, reduced p-tau and ß-amyloid (Aß) peptide-stained neurons, and improved memory. The BBB-bypassing Nrf2-activating polysaccharide reported here may be effective in reducing oxidative stress and neurodegeneration in AD.

Peptide Nanotube Encapsulated Enzyme Biosensor for Vapor Phase Detection of Malathion, an Organophosphorus Compound.

This study explores the use of a butyrylcholinesterase (BChE)-based, reversible reaction biosensor using screen-printed electrodes (SPEs) having a smaller working surface area than the single-use electrodes previously studied. Previous research demonstrated the prospective application of a single-use biosensor fabricated with an acetylcholinesterase (AChE) enzyme encapsulated in peptide nanotubes (PNTs) and enhanced with horseradish peroxidase (HRP) to detect organophosphorus compounds (OPCs) in aqueous and gas phases. In the current study, potential improvements to the biosensor are investigated. BChE-based biosensors were fabricated using PNTs, HRP, and Nafion in combination to increase the reactive surface area, enhance sensitivity, and maintain enzyme stability. Cyclic voltammetry (CV) was used along with the new modified sensor to measure malathion concentration in the gas phase. The results show that a BChE-based biosensor could reliably measure gas phase malathion concentrations between 6-25 ppbv by CV with the extent of inhibition linearly proportional to the malathion concentration (R2 = 0.941). This research demonstrated that fabricated BChE-based biosensors could be stored without cold storage requirement for up to six weeks with minimal performance degradation. Moreover, the sensor electrodes were each reused several times, and were still useable at the conclusion of the research. This research demonstrates the potential of fabricating a reusable, inexpensive biosensor that is capable of OPC detection with high sensitivity and a low detection limit without a long-term cold storage requirement.

Revaprazan-loaded surface-modified solid dispersion: physicochemical characterization and in vivo evaluation.

The purpose of this research was to develop a novel revaprazan-loaded surface-modified solid dispersion (SMSD) with improved drug solubility and oral bioavailability. The impact of carriers on aqueous solubility of revaprazan was investigated. HPMC and Cremophor A25 were selected as an appropriate polymer and surfactant, respectively, due to their high drug solubility. Numerous SMSDs were prepared with various concentrations of carriers, using distilled water, and the drug solubility of each was assessed. Moreover, the physicochemical properties, dissolution and pharmacokinetics of selected SMSD in rats were assessed in comparison to revaprazan powder. Of the SMSDs assessed, the SMSD composed of revaprazan/HPMC/Cremophor A25 at the weight ratio of 1:0.28:1.12 had the most enhanced drug solubility (∼6000-fold). It was characterized by particles with a relatively rough surface, suggesting that the carriers were attached onto the surface of the unchanged crystalline revaprazan powder. It had a significantly higher dissolution rate, AUC and Cmax, and a faster Tmax value in comparison to revaprazan powder, with a 5.3-fold improvement in oral bioavailability of revaprazan. Therefore, from an environmental perspective, this SMSD system prepared with water, and without organic solvents, should be recommended as a revaprazan-loaded oral pharmaceutical alternative.

Novel revaprazan-loaded gelatin microsphere with enhanced drug solubility and oral bioavailability.

To develop a novel revaprazan-loaded gelatine microsphere with enhanced solubility and oral bioavailability, numerous gelatine microspheres were prepared using a spray-drying technique. The impact of gelatine amount on drug solubility in the gelatine microspheres was investigated. The physicochemical properties of the selected gelatine microsphere, such as shape, particle size and crystallinity, were evaluated. Moreover, its dissolution and pharmacokinetics in rats were assessed in comparison with revaprazan powder. Amongst the gelatine microspheres tested, the gelatine microsphere consisting of revaprazan and gelatine (1:2, w/w), which gave about 150-fold increased solubility, had the most enhanced drug solubility. It provided a spherical shape, amorphous drug and reduced particle size. Furthermore, it gave a higher dissolution rate and plasma concentration than did revaprazan powder. Particularly, it gave about 2.3-fold improved oral bioavailability in comparison with revaprazan powder. Therefore, this novel gelatine microsphere system is recommended as an oral pharmaceutical product of poorly water-soluble revaprazan.

Development of a novel celecoxib-loaded nanosuspension using a wet media milling process.

To develop a novel celecoxib (CXB)-loaded drug delivery system, numerous nanosuspensions were prepared with various polymers and surfactants using a wet media milling process, and their particle sizes were subsequently determined. A 24 full factorial design was used to identify the most appropriate preparation conditions. Pharmacokinetics of the selected nanosuspension were performed in rats and compared with those of a drug powder and a commercial CXB-loaded product. Among the carriers investigated, copovidone and sodium lauryl sulphate gave the smallest particle size of the drug in the nanosuspension. In particular, the nanosuspension prepared with 5% CXB, 4% copovidone, and 0.1% sodium lauryl sulphate, under the appropriate conditions, showed a particle size of approximately 190 nm, which was physically stable for at least 8 weeks. This nanosuspension provided a significantly higher plasma concentration and AUC in rats as compared with the drug powder and the commercial product. Thus, this novel CXB-loaded nanosuspension is a promising candidate with excellent stability and enhanced oral bioavailability.

Life cycle assessment of superheated steam drying technology as a novel cow manure management method.

Common methods of managing dairy manure are directly applying it to the farm field as a fertilizer. For direct application without any type of treatment, the majority of nutrients in the manure run off to the local river and lake during precipitation periods. The algae bloom is one of the environmental outcomes due to eutrophication of the lakes, which may jeopardize the quality of drinking water. In this study, superheated steam drying (SSD) technology is investigated as an alternative manure management method. Rapidly dried cow manure can be used as alternative fuel. Evaluations of energy payback time (EPBT) and life cycle assessment (LCA) of the SSD technology are presented in the SSD scenario and the results are compared with those of the direct field application (FA) of fresh manure and anaerobic digestion (AD). The heat required for the generation of superheated steam in the SSD scenario is provided from combustion of the dry manure to reduce energy costs. The results for the SSD process show 95% and 70% lower eutrophication and global warming potential in comparison to the FA scenario. Acidification potential for SSD turned out to be 35% higher than FA. The comparison of SSD with AD for their EPBT and normalized impacts indicated that the proposed SSD scenario has higher environmental sustainability than AD (70% lower impact), and is likely an economically better choice compared to conventional AD method (87% lower EPBT) for the future investment.

Influence of polyvinylpyrrolidone quantity on the solubility, crystallinity and oral bioavailability of fenofibrate in solvent-evaporated microspheres.

The objective of this study is to explore the influence of polyvinylpyrrolidone (PVP) quantity on the solubility, crystallinity and oral bioavailability of poorly water-soluble fenofibrate in solvent-evaporated microspheres. Numerous microspheres were prepared with fenofibrate, sodium lauryl sulphate (SLS) and PVP using the spray-drying technique. Their aqueous solubility, dissolution, physicochemical properties and pharmacokinetics in rats were assessed. The drug in the solvent-evaporated microspheres composed of fenofibrate, PVP and SLS at the weight ratio of 1:0.5:0.25 was not entirely changed to the amorphous form and partially in the microcrystalline state. However, the microspheres at the weight ratio of 1:4:0.25 provided the entire conversion to the amorphous form. The latter microspheres, with an improvement of about 115 000-fold in aqueous solubility and 5.6-fold improvement in oral bioavailability compared with the drug powder, gave higher aqueous solubility and oral bioavailability compared with the former. Thus, PVP quantity played an important role in these properties of fenofibrate in the solvent-evaporated microspheres.

Reduced Glutathione-Modified Electrode for the Detection of Hydroxyl Free Radicals.

Hydroxyl radicals (•OH) are known as essential chemicals for cells to maintain their normal functions and defensive responses. However, a high concentration of •OH may cause oxidative stress-related diseases, such as cancer, inflammation, and cardiovascular disorders. Therefore, •OH can be used as a biomarker to detect the onset of these disorders at an early stage. Reduced glutathione (GSH), a well-known tripeptide for its antioxidant capacity against reactive oxygen species (ROS), was immobilized on a screen-printed carbon electrode (SPCE) to develop a real-time detection sensor with a high selectivity towards •OH. The signals produced by the interaction of the GSH-modified sensor and •OH were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The CV curve of the GSH-modified sensor in the Fenton reagent exhibited a pair of well-defined peaks, demonstrating the redox reaction of the electrochemical sensor and •OH. The sensor showed a linear relationship between the redox response and the concentration of •OH with a limit of detection (LOD) of 49 µM. Furthermore, using EIS studies, the proposed sensor demonstrated the capability of differentiating •OH from hydrogen peroxide (H2O2), a similar oxidizing chemical. After being immersed in the Fenton solution for 1 hr, redox peaks in the CV curve of the GSH-modified electrode disappeared, revealing that the immobilized GSH on the electrode was oxidized and turned to glutathione disulfide (GSSG). However, it was demonstrated that the oxidized GSH surface could be reversed back to the reduced state by reacting with a solution of glutathione reductase (GR) and nicotinamide adenine dinucleotide phosphate (NADPH), and possibly reused for •OH detection.

Polysaccharide-Based Composite Scaffolds for Osteochondral and Enthesis Regeneration.

The rotator cuff and Achilles tendons along with the anterior cruciate ligament (ACL) are frequently injured with limited healing capacity. At the soft-hard tissue interface, enthesis is prone to get damaged and its regeneration in osteochondral defects is essential for complete healing. The current clinical techniques used in suturing procedures to reattach tendons to bones need much improvement for the generation of the native interface tissue, that is, enthesis, for patients to regain their full functions. Recently, inspired by the composite native tissue, much effort has been made to fabricate composite scaffolds for enthesis tissue regeneration. This review first focuses on the studies that used composite scaffolds for the regeneration of enthesis. Then, the use of polysaccharides for osteochondral tissue engineering is reviewed and their potential for enthesis regeneration is presented based on their supporting effects on osteogenesis and chondrogenesis. Gellan gum (GG) is selected and reviewed as a promising polysaccharide due to its unique osteogenic and chondrogenic activities that help avoid the inherent weakness of dissimilar materials in composite scaffolds. In addition, original preliminary results showed that GG supports collagen type I production and upregulation of osteogenic marker genes. Impact Statement Enthesis regeneration is essential for complete and functional healing of tendon and ligament tissues. Current suturing techniques to reattach the tendon/ligament to bones have high failure rates. This review highlights the studies on biomimetic scaffolds aimed to regenerate enthesis. In addition, the potential of using polysaccharides to regenerate enthesis is discussed based on their ability to regenerate osteochondral tissues. Gellan gum is presented as a promising biopolymer that can be modified to simultaneously support bone and cartilage regeneration by providing structural continuity for the scaffold.

Rheological properties and decomposition rates of Gellan gum/hyaluronic acid/ß-tricalcium phosphate mixtures.

The effects of ß-tricalcium phosphate (TCP) on the mixture of low acyl gellan gum (LA-GAGR) and hyaluronic acid (HA) were investigated for the rheological properties and decomposition rates. All the tested mixture samples exhibited shear-thinning and typical viscoelastic behaviors. The sample made with 1.0% TCP and 0.30% LA-GAGR had the highest viscosity and loss and storage moduli and displayed gel-like behavior with the highest swelling capacity. The same mixture also exhibited the lowest average cumulative decomposition rate. High concentrations of LA-GAGR and TCP increased the degree of cross-linking of the polysaccharides, and as a result, the mixture was more elastic and less fluidic and decomposed slower. The samples prepared by gradual mixing of LA-GAGR and TCP decomposed slower than the sample prepared by sudden mixing, which indicates the well-dispersed TCP enhanced cross-linking of the polymers. This study demonstrates the possible applicability of natural polysaccharide-based shear-thinning gels for biomedical applications.

Detection Technologies for Reactive Oxygen Species: Fluorescence and Electrochemical Methods and Their Applications.

Reactive oxygen species (ROS) have been found in plants, mammals, and natural environmental processes. The presence of ROS in mammals has been linked to the development of severe diseases, such as diabetes, cancer, tumors, and several neurodegenerative conditions. The most common ROS involved in human health are superoxide (O2•-), hydrogen peroxide (H2O2), and hydroxyl radicals (•OH). Organic and inorganic molecules have been integrated with various methods to detect and monitor ROS for understanding the effect of their presence and concentration on diseases caused by oxidative stress. Among several techniques, fluorescence and electrochemical methods have been recently developed and employed for the detection of ROS. This literature review intends to critically discuss the development of these techniques to date, as well as their application for in vitro and in vivo ROS detection regarding free-radical-related diseases. Moreover, important insights into and further steps for using fluorescence and electrochemical methods in the detection of ROS are presented.

Molecular weight effects of low acyl gellan gum on antioxidant capacity and rheological properties.

The current study investigated the antioxidant capacity of enzymatically cleaved low acyl gellan gum (LA-GAGR) fragments, named midi-GAGR (MWv : 1.2 × 105  Da) and mini-GAGR (MWv : 2.5 × 104  Da). Three different methods-hydroxide assay, superoxide assay, and DPPH assay-were used to measure the antioxidant capacity of the low acyl gellan gum fragments. Both mini-GAGR and midi-GAGR showed similar antioxidant capacities, 27.1% and 25.6%, respectively, for hydroxide radicals, whereas ascorbic acid showed 9.8%. For superoxide radicals, the fragments scavenged 41.7% (mini) and 35.6% (midi) of free radicals compared to 10.6% removal by ascorbic acid. Mini- and midi-GAGR displayed modest scavenging capabilities with DPPH radicals (8.5% and 6.6%, respectively) as compared to ascorbic acid (96.3%). Both midi- and mini-GAGR showed less gel-like behaviors than LA-GAGR. Midi-GAGR was observed to have a transition from liquid to gel at 63 rad/s. PRACTICAL APPLICATION: The results in the manuscript are helpful when gellan gum and its derivatives are directly applied to food processing as a dietary fiber supplement or a stabilizer for functional beverages. The antioxidant capacity results can be used to promote the functionality of gellan gum as a food additive and for controlling cell adhesion and growth on gellan gum scaffolds. The rheology results will be useful for synthesis of scaffolds for bone tissue generation and facilitating clinical treatments when gellan gum is injected as an adsorbent or a filler for treating bone fractures. In the pharmaceutical industry, they are useful when controlling the therapeutic effects of drug delivery systems.

Supersensitive CeOx-based nanocomposite sensor for the electrochemical detection of hydroxyl free radicals.

It is well known that an excess of hydroxyl radicals (˙OH) in the human body is responsible for oxidative stress-related diseases. An understanding of the relationship between the concentration of ˙OH and those diseases could contribute to better diagnosis and prevention. Here we present a supersensitive nanosensor integrated with an electrochemical method to measure the concentration of ˙OH in vitro. The electrochemical sensor consists of a composite comprised of ultrasmall cerium oxide nanoclusters (<2 nm) grafted to a highly conductive carbon deposited on a screen-printed carbon electrode (SPCE). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to analyze the interaction between cerium oxide nanoclusters and ˙OH. The CV results demonstrated that this electrochemical sensor had the capacity of detecting ˙OH with a high degree of accuracy and selectivity, achieving a consistent performance. Additionally, EIS results confirmed that our electrochemical sensor was able to differentiate ˙OH from hydrogen peroxide (H2O2), which is another common reactive oxygen species (ROS) found in the human body. The limit of detection (LOD) observed with this electrochemical sensor was of 0.6 µM. Furthermore, this nanosized cerium oxide-based electrochemical sensor successfully detected in vitro the presence of ˙OH in preosteoblast cells from newborn mouse bone tissue. The supersensitive electrochemical sensor is expected to be beneficially used in multiple applications, including medical diagnosis, fuel-cell technology, and food and cosmetic industries.

Comparative study between high-pressure homogenisation and Shirasu porous glass membrane technique in sildenafil base-loaded solid SNEDDS: Effects on physicochemical properties and in vivo characteristics.

The purpose of this study was to compare two types of emulsification techniques in a solid self-nanoemulsifying drug delivery system (SNEDDS); high-pressure homogenisation (HPH) and Shirasu porous glass membrane (SPG). Those two emulsification processes enhanced the solubility, dissolution and oral bioavailability of poorly water-soluble sildenafil base (SB) by producing fine and well-dispersed nanoemulsion droplet. The liquid SNEDDS consisting of Labrasol/Transcutol HP/coconut oil at the weight of 72/18/10, gave the smallest emulsion droplet size among the prepared liquid SNEDDS formulations. Then, the SB-loaded liquid SNEDDS was dissolved in the deionised water and applied to HPH or SPG techniques. Aerosil 200 was suspended as a mesoporous carrier and spray-dried, producing an SB-loaded solid SNEDDS. The emulsion droplet size, solubility and dissolution of each emulsification process were compared to the solid SNEDDS fabricated without any treatment of additional emulsification. Moreover, the physicochemical properties of all formulations were compared. The crystalline state of the drug in all products was converted to the amorphous state. The solid SNEDDS, subjected to HPH technique, provided fine and well-dispersed nanoemulsion. Additionally, it increasingly improved the drug solubility and dissolution as compared to the others, including SB powder, non-treated (NT) and SPG. Furthermore, it gave improved Cmax and increased AUC compared to SB powder and SPG, indicating HPH enhanced the oral bioavailability of SB the most. Thus, this solid SNEDDS with HPH would be strongly suggested as an oral SB-loaded pharmaceutical product.

Comparison of Three Different Aqueous Microenvironments for Enhancing Oral Bioavailability of Sildenafil: Solid Self-Nanoemulsifying Drug Delivery System, Amorphous Microspheres and Crystalline Microspheres.

BACKGROUND: The purpose of this study was to screen various drug delivery systems for improving the aqueous solubility and oral bioavailability of sildenafil. Three representative techniques, solid self-nanoemulsifying drug delivery systems (SNEDDS), amorphous microspheres and crystalline microspheres, were compared. METHODS: Both microspheres systems contained sildenafil:Labrasol:PVP at a weight ratio of 1:1:6. The amorphous microspheres were manufactured using ethanol, while crystalline microspheres were generated using distilled water. Liquid SNEDDS was composed of sildenafil:Labrasol:Transcutol HP:Captex 300 in the ratio of 1:70:15:15 (w:w:w:w). The solidification process in SNEDDS was performed using HDK N20 Pharma as a solid carrier. RESULTS: The amorphous microspheres appeared spherical with significantly decreased particle size compared to the drug powder. The crystalline microspheres exhibited a rough surface with no major particle-size difference compared with sildenafil powder, indicating that the hydrophilic excipients adhered to the sildenafil crystal. Solid SNEDDS presented a smooth surface, assuming that the oily liquid was adsorbed to the porous solid carrier. According to the physicochemical evaluation, the crystalline state maintained in crystalline microspheres, whereas the crystal state changed to amorphous state in other formulations. Amorphous microspheres, crystalline microspheres and solid SNEDDS produced about 79, 55, 82-fold increased solubility, compared to drug powder. Moreover, the prepared formulations provided a higher dissolution rate (%) and plasma concentration than did the drug powder (performance order; solid SNEDDS ≥ amorphous microspheres ≥ crystalline microspheres > drug powder). Among the formulations, solid SNEDDS demonstrated the highest improvement in oral bioavailability (AUC; 1508.78 ± 343.95 h·ng/mL). CONCLUSION: Therefore, solid SNEDDS could be recommended as an oral dosage form for enhancing the oral bioavailability of sildenafil.

Detection of Hydroxyl Radicals Using Cerium Oxide/Graphene Oxide Composite on Prussian Blue.

A composite sensor consisting of two separate inorganic layers of Prussian blue (PB) and the composite of cerium oxide nanoparticles (CeNPs) and graphene oxide (GO), is tested with •OH radicals. The signals from the interaction between the composite layers and •OH radicals are characterized using cyclic voltammetry (CV). The degradation of PB in the presence of H2O2 and •OH radicals is observed and its impact on the sensor efficiency is investigated. The results show that the composite sensor differentiates between the solutions with and without •OH radicals by the increase of oxidation current in the presence of •OH radicals. The redox response shows a linear relation with the concentration of •OH radicals where the limit of detection, LOD, is found at 60 µM (100 µM without the PB layer). When additional composite layers are applied on the composite sensor to prevent the degradation of PB layer, the PB layer is still observed to be degraded. Furthermore, the sensor conductivity is found to decrease with the additional layers of composite. Although the CeNP/GO/PB composite sensor demonstrates high sensitivity with •OH radicals at low concentrations, it can only be used once due to the degradation of PB.

Effects of phenol and natural phenolic compounds on biofilm formation by Pseudomonas aeruginosa.

A biofilm is formed as a result of adhesion of microorganisms to various surfaces with the production of extracellular polymers (polysaccharides and proteins). Biofilms cause serious problems in the chemical, medical and pharmaceutical industries. Recent findings indicate that some natural phenolic compounds found in plants have an anti-biofouling effect on biofilm formation by Gram-negative bacteria. The anti-biofouling activities of 14 selected phenol and natural phenolic compounds were tested against Pseudomonas aeruginosa, using a microtiter-plate. A modified microtiter-plate assay was used because it enabled indirect measurement of bacterial cells attached to the surface of the wells. This assay involved fixing the bacterial film with methanol, staining with crystal violet dye and then releasing the bound dye with 33% glacial acetic acid. The optical density (OD) of the solution was measured at 570 nm by using an automated ICN Flow Titertek Multiscan Plus reader. Phenol and natural phenolic compounds except ethyl linoleate and tocopherol showed a significant reduction in biofilm formation by P. aeruginosa.