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The development of biofilm by pure SRB culture on carbon steel, and its role on corrosion were investigated using microscopic, spectroscopic, electrochemical and surface characterization techniques. Tubercle biofilm and irregularly shaped pits were observed on steel surfaces in high-nutrient biotic solution. Owing to development of a protective FeS film in 72 h immersion, corrosion resistance improved. In nutrient-deficient medium, a greater bacterial density attached to the metal surface as the consequence of starved bacteria seeking energy sources from metal. However, electrochemical non-homogeneity developed at the locations of their attachment, that gradually grew over the entire surface. [ABSTRACT FROM AUTHOR]

Frost formation poses challenges in various sectors, impacting efficiency and safety. Active methods involve energy-consuming processes like heating, while passive approaches utilize protective coatings or hydrophobic surfaces to minimize frost aggregation. However, low thermal conductivity remains a concern. In previous research, lotus leaves inspired micro-meter-sized asperities were effective in controlling frost growth. Applying this concept to plastic, particularly low-cost PVC, raised challenges due to its low thermal conductivity. The study explores utilizing carbon black (CB) waste from hard coating processes to enhance PVC's thermal conductivity, forming a micro-plastic texture (MPT). The research investigates the influence of CB amount and AC voltage frequency on MPT features and thermal conductivity, aiming for a sustainable solution to address frost-related issues. Incorporating 0.9 vol.% CB into PVC solution under a 2 kV AC electric field (6 Hz) resulted in an extensively distributed MPT on the surface. This MPT, formed under the 6 Hz condition, exhibited no in-plane bias, making it an ideal surface for preventing frost formation. Adding CB to PVC within the range of 0 to 2.1 vol.% and applying a 2 kV, 6 Hz AC electric field led to independent MPT formation at concentrations between 0.4 to 1.2 vol.%. Conducting cooling tests in atmospheric and vacuum conditions, we found that MPT with 0.9 vol.% CB displayed a thermal conductivity of approximately 0.19 kW/mK in atmospheric conditions, similar to aluminum alloy. However, with 1.2 vol.% or more CB, it decreased to about 0.15 kW/mK. In vacuum conditions, MPT with 1.2 vol.% CB exhibited a higher thermal conductivity of 0.063 kW/mK compared to the 0.055 kW/mK observed in MPT with 0 vol.% CB.

The development of biofilm by pure SRB culture on carbon steel, and its role on corrosion were investigated using microscopic, spectroscopic, electrochemical and surface characterization techniques. Tubercle biofilm and irregularly shaped pits were observed on steel surfaces in high-nutrient biotic solution. Owing to development of a protective FeS film in 72 h immersion, corrosion resistance improved. In nutrient-deficient medium, a greater bacterial density attached to the metal surface as the consequence of starved bacteria seeking energy sources from metal. However, electrochemical non-homogeneity developed at the locations of their attachment, that gradually grew over the entire surface.

Oxalic acid (C2H2O4) and Calcium (Ca2+) react to form the salt Calcium oxalate (CaOx), which crystallises into a variety of topologically diverse crystals. CaOx crystals have been found in at least 215 plant groups, which corresponds to numerous species. Crystals can be found in vascular, epidermal, ground, and other tissues in addition to roots, stems, leaves, flowers, fruits, and seeds. They develop in crystal idioblasts, specialised cells, in their vacuoles. According to recent studies, CaOx crystals are in fact useful tools that are crucial, especially in stressful conditions. As plants lack an excretory system, the Ca component regulates the cytosolic concentration levels and immobilises excess amounts of this element. Oxalates operate as a dynamic carbon store and set off an alert during photosynthesis, which results in the production of CO2. The article aims to provide readers with a greater understanding of Portulacaria afra's CaOx crystals and the projected crystal disintegration that would liberate carbon and supply the photosynthetic cycles with it as defence against salinity and drought stress.

Heterogeneous nucleation of water vapor on particles to promote particulate abatement is widely used in the fields of industrial dust removal pretreatment technology. Water vapor competition is a very important phenomenon in the process of particles nucleation, but due to the limitations of experimental methods, this phenomenon is difficult to observe and rarely received attention. Therefore, in order to reveal the influence of water vapor competition on particles nucleation, a direct visualization method based on the Environmental Scanning Electron Microscope (ESEM) is proposed in this work to obtain the water vapor competition process during particles nucleation at a microscopic scale. Firstly the effect of particle's surface structure on water vapor competition in heterogeneous nucleation of water vapor on the particles is visualized by the ESEM. The results show that the embryo appears randomly at any edge of the smooth spherical particle because water vapor competition at each point on the particle's surface is equal. While the embryo always appears on the protuberance of the convex spherical particle since water vapor competition is strongest there. Then the influence of particle's surface energy on water vapor competition is also visualized. Water vapor preferentially nucleates on the particle with smaller contact angle, which means the higher the surface energy, the more conducive it is for water vapor competition to form embryos. Finally, the impact of particle's aggregation modes on water vapor competition is also visualized by the ESEM. The results show that when the vapor supersaturation is constant, water vapor only nucleates on the two-close and three-circled particles while particles nucleation on the single particle is never observed. This is due to the strong water vapor competition at the particle's junction, where the embryos will collide and coalesce to form an embryo larger than the critical size to activate particles nucleation. The results of this study will deepen the microscopic understanding of water vapor competition in the process of particles nucleation. [ABSTRACT FROM AUTHOR]

This study is part of a comprehensive investigation that was performed in regard to a case of alterations on a carbographic ribbon used in a typewriter that was found and seized by inner security operations of the Arma dei Carabinieri, Italy. Thirty‐six coded scripts possessing potentially and criminally liable content were present on the tape; however, only the 6th and 7th scripts exhibited alterations of an uncertain nature. The study included sampling that was performed under sterile conditions of a large surface area of carbographic ribbons. A protocol based on physico‐chemical, microbiological, and biomolecular tools was established. Preliminary results revealed the presence of fungal contamination that was primarily located on the inner surface of the 6th and 7th scripts on the black carbographic ribbon. One fungal strain was isolated and identified by universal ITS‐PCR primer and rDNA sequencing as Alternaria infectoria strain NIS4. Fungal growth was monitored for 3 weeks in the laboratory under different environmental conditions (temperature, open‐closed system, and substrate). The A. infectoria NIS4 strain exhibited the best growth at 28°C under a closed system with RH near saturation. We also noted that the fungal growth was abundant at 15°C. Moreover, this fungus (a potential human pathogen) possessed the ability to colonize the surface of the new carbographic ribbon even when using mineral medium; however, this only occurred in a closed system environment and not in open systems due to rapid desiccation. Under our experimental conditions, the A. infectoria NIS4 strain could degrade gelatin as an organic matter present in trace amounts that are often used as a binder in a carbographic ribbon emulsions. The results revealed that the isolated microorganism was the major biological candidate capable of altering the investigated carbographic ribbon; however, these alterations could only occur under favourable environmental conditions. Aims: Identify the cause of microbial alterations on a carbographic ribbon in a typewriter used in a hypogean Italian criminal house named "covo." Methods and Results: The isolation and identification of biodeteriogens (Alternaria infectoria NIS4) were performed using both culture‐dependent and‐independent methods, including ITS regions‐primed PCR and rDNA techniques. Environmental scanning electron microscopy (ESEM) and optical observations were also performed. Growth tests and biodeterioration simulation tests on carbographic ribbons at the lab scale were performed under different environmental conditions. The A. infectoria NIS4 strain exhibited biodeterioration activity on carbographic ribbons under environmental conditions that were extremely favourable for growth. A high ability to colonize carbographic ribbon surfaces with fast and abundant growth at both 15°C and 28°C under lab‐scale conditions at RH near saturation was observed. Conclusions: In this forensic case study, the ability of the isolated micromycetes A. infectoria NIS4 strain to colonize and induce alterations and degradation in a carbographic ribbon stored under indoor environmental conditions was examined. When favourable conditions change over time, the risk of microbial colonization and the damage produced by the fungal biodeterioration processes on the synthetic material objects has been confirmed. Significance and Impact of Study: The current study contributes to the knowledge of biodeterioration processes in carbographic ribbon and the responsible agents, and our study provides an example of how environmental microbiology can also aid in forensic studies. [ABSTRACT FROM AUTHOR]

Microscopic, electrochemical and surface characterization techniques were used to investigate the effects of iron reducing bacteria (IRB) biofilm on carbon steel corrosion for 72 and 168 h under batch conditions. The organic nutrient availability for the bacteria was varied to evaluate biofilms formed under nutritionally rich, as compared to nutritionally deficient, conditions. Focused ion beam-scanning electron microscopy (FIB-SEM) was used to investigate the effect of subsurface biofilm structures on the corrosion characteristics of carbon steel. Hydrated biofilms produced by IRB were observed under environmental scanning electron microscope (ESEM) with minimal surface preparation, and the elemental composition of the biofilms was investigated using energy dispersive spectroscopy (EDX). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) was used to provide information on the organic and inorganic chemical makeup of the biofilms. Electrochemical techniques employed for assessing corrosion, by open circuit potential, linear polarization and potentiodynamic polarization tests indicated no significant difference in the corrosion resistance for carbon steel in IRB-inoculated, compared to the abiotic solutions of common Postgate C after 72 and 168 h. However, the steel was found to be more susceptible to corrosion when the yeast extract was removed from the biotic environment for the 168 h test. In the absence of yeast nutrient, it is postulated that IRB received energy by transforming the protective film of Fe3+ into more soluble Fe2+ products. [ABSTRACT FROM AUTHOR]

The clinker mineral tricalcium aluminate (C3A) determines largely the early hydration characteristics of Portland cement. Therefore, understanding the influence of the polymer on the hydration of C3A is contributed to clarify the mechanism of polymer on the early hydration of cement-based material. The hydration environment of C3A in Portland cement was simulated by using C3A mineral, gypsum ( C S ¯ · H 2 ) and calcium hydroxide (CH). The specimens, whose water cement ratio (mw/mc) was 0.4 and carboxylated styrene–butadiene (SB) copolymer cement ratio (mp/mc) was 0, 5, 10, 15 and 20%, were cured at 20 ℃/RH90% for 6 h, 12 h, 1 day, 3 days, 7 days and 28 days, respectively. The effect of SB copolymer on the system of C3A– C S ¯ · H 2 –CH was investigated by the combination of isothermal calorimetry, X-ray diffraction (XRD) and environmental scanning electron microscopy (ESEM). It was found that the SB copolymer retards C3A hydration and reduces the heat of hydration. The XRD results revealed that addition of SB copolymer inhibits the consumption of C3A, delays the conversion of ettringite (AFt) to calcium monosulfate aluminate hydrate (AFm), promotes the formation of hydrated calcium aluminate (C4AH13), and reduces the crystallinity of AFm. [ABSTRACT FROM AUTHOR]

Microscopic, electrochemical and surface characterization techniques were used to investigate the effects of iron reducing bacteria (IRB) biofilm on carbon steel corrosion for 72 and 168 h under batch conditions. The organic nutrient availability for the bacteria was varied to evaluate biofilms formed under nutritionally rich, as compared to nutritionally deficient, conditions. Focused ion beam-scanning electron microscopy (FIB-SEM) was used to investigate the effect of subsurface biofilm structures on the corrosion characteristics of carbon steel. Hydrated biofilms produced by IRB were observed under environmental scanning electron microscope (ESEM) with minimal surface preparation, and the elemental composition of the biofilms was investigated using energy dispersive spectroscopy (EDX). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) was used to provide information on the organic and inorganic chemical makeup of the biofilms. Electrochemical techniques employed for assessing corrosion, by open circuit potential, linear polarization and potentiodynamic polarization tests indicated no significant difference in the corrosion resistance for carbon steel in IRB-inoculated, compared to the abiotic solutions of common Postgate C after 72 and 168 h. However, the steel was found to be more susceptible to corrosion when the yeast extract was removed from the biotic environment for the 168 h test. In the absence of yeast nutrient, it is postulated that IRB received energy by transforming the protective film of Fe3+ into more soluble Fe2+ products.

PREMISE: Biological invasions increasingly threaten native biodiversity and ecosystem services. One notable example is the common reed, Phragmites australis, which aggressively invades North American salt marshes. Elevated atmospheric CO2 and nitrogen pollution enhance its growth and facilitate invasion because P. australis responds more strongly to these enrichments than do native species. We investigated how modifications to stomatal features contribute to strong photosynthetic responses to CO2 and nitrogen enrichment in P. australis by evaluating stomatal shifts under experimental conditions and relating them to maximal stomatal conductance (gwmax) and photosynthetic rates. METHODS: Plants were grown in situ in open‐top chambers under ambient and elevated atmospheric CO2 (eCO2) and porewater nitrogen (Nenr) in a Chesapeake Bay tidal marsh. We measured light‐saturated carbon assimilation rates (Asat) and stomatal characteristics, from which we calculated gwmax and determined whether CO2 and Nenr altered the relationship between gwmax and Asat. RESULTS: eCO2 and Nenr enhanced both gwmax and Asat, but to differing degrees; gwmax was more strongly influenced by Nenr through increases in stomatal density while Asat was more strongly stimulated by eCO2. There was a positive relationship between gwmax and Asat that was not modified by eCO2 or Nenr, individually or in combination. CONCLUSIONS: Changes in stomatal features co‐occur with previously described responses of P. australis to eCO2 and Nenr. Complementary responses of stomatal length and density to these global change factors may facilitate greater stomatal conductance and carbon gain, contributing to the invasiveness of the introduced lineage. [ABSTRACT FROM AUTHOR]

Processes of water condensation and desublimation on solid surfaces are ubiquitous in nature and essential for various industrial applications, which are crucial for their performance. Despite their significance, these processes are not well understood due to the lack of methods that can provide insight at the nanolevel into the very first stages of phase transitions. Taking advantage of synchrotron grazing-incidence wide-angle X-ray scattering (GIWAXS) and environmental scanning electron microscopy (ESEM), two pathways of the frosting process from supersaturated vapors were studied in real time for substrates with different wettabilities ranging from highly hydrophilic to superhydrophobic. Within GIWAXS, a fully quantitative structural and orientational characterization of the undergoing phase transition reveals the information on degree of crystallinity of the new phase and determines the ordering at the surfaces and inside the films at the initial stages of water/ice nucleation from vapor onto the substrates. The diversity of frosting scenarios, including direct desublimation from the vapor and two-stage condensation-freezing processes, was observed by both GIWAXS and ESEM for different combinations of substrate wettability and vapor supersaturations. The classical nucleation theory straightforwardly predicts the pathway of the phase transition for hydrophobic and superhydrophobic substrates. The case of hydrophilic substrates is more intricate because the barriers in Gibbs free energy for nucleating both liquid and solid embryos are close to each other and comparable to thermal energy k B T . At that end, classical nucleation theory allows concluding a relation between contact angles for ice and water embryos on the basis of the observed frosting pathway.

Heterogeneous condensation of water vapor on particles is widely used in the fields of atmospheric cloud physics and industrial particulate abatement. Particles surface wettability plays a fundamental role in water vapor condensation, but a microscopic insight of the impact of particles surface wettability on vapor condensation is lacked. Therefore, the Environmental Scanning Electron Microscope (ESEM) is adopted to directly visualize vapor condensation on particles with different surface wettability at a microscopic scale. Firstly heterogeneous condensation of water vapor on the hydrophilic and hydrophobic particles is obtained by the ESEM. The results show that when water vapor condenses on the hydrophilic particles, a spherical cap-shaped embryo first appears and subsequently develops into a spherical droplet to finally wrap the single particle. While on the hydrophobic particles, a spherical cap-shaped embryo will first appear and continue to grow into a spherical droplet to finally detach from the particle. Then the wetting coefficient is introduced to characterize the wetting degree of the droplet on the particle. The wetting coefficient will increase with the decrease of the contact angle, so the wettability of water vapor on the hydrophilic particles is better than that on the hydrophobic particles. Meanwhile based on classical nucleation theory, the geometrical factor and the critical supersaturation will decrease when the contact angle decreases, making it easier for water vapor to condense on the hydrophilic particles. Finally the line tension is investigated to explain the condensation mechanism. On the hydrophilic particles, when the droplet has crossed the equatorial line of the particle, the positive line tension promotes the spreading movement of the droplet over the particle to form a wrapped spherical droplet. Therefore, there is a transition from a spherical cap-shaped embryo before crossing the equatorial line of the particle to a wrapped spherical droplet after crossing the equatorial line. While on the hydrophobic particles, the negative line tension suppresses the spreading movement of the droplet over the particle to form a detached spherical droplet. So there is a transition from a spherical cap-shaped embryo before crossing the equatorial line of the particle to a detached spherical droplet after crossing the equatorial line. • Heterogeneous condensation of water vapor on hydrophilic and hydrophobic particles is directly visualized. • A spherical cap-shaped embryo appears on the hydrophilic particle and develops into a spherical droplet to wrap the particle. • A spherical cap-shaped embryo appears on the hydrophobic particle and grows into a spherical droplet to detach from the particle. • The positive line tension promotes the spreading movement of the droplet over the hydrophilic particle. • The negative line tension suppresses the spreading movement of the droplet over the hydrophobic particle. [ABSTRACT FROM AUTHOR]

Results of positron annihilation lifetime spectroscopy (PALS) and microscopic studies on simple microorganisms, brewing yeasts, are presented. Lifetime of ortho-positronium (o-Ps) were found to change from 2.4 to 2.9 ns (longer-lived component) for lyophilized and aqueous yeasts, respectively. Also hygroscopicity of yeasts in time was examined, allowing to check how water – the main component of the cell – affects PALS parameters, thus lifetime of o-Ps were found to change from 1.2 to 1.4 ns (shorter-lived component) for the dried yeasts. The time sufficient to hydrate the cells was found below 10 hours. In the presence of liquid water, an indication of reorganization of yeast in the molecular scale was observed. Microscopic images of the lyophilized, dried, and wet yeasts with best possible resolution were obtained using inverted microscopy (IM) and environmental scanning electron microscopy (ESEM) methods. As a result, visible changes to the surface of the cell me mbrane were observed in ESEM images.

Diatomite, a rock formed by the accumulation of opaline diatom frustules, is a preferred rawmaterial for themanufacturing of filters. Its uniqueness relies on the high porosity and inertness of the frustules. The presence of carbonates in some diatomite ores hinders these properties. The purpose of this study was to identify the type of carbonates and their association with the ore in a diatomite deposit, and to assess the suitability of determining the quality of the ore using techniques with potential for in-pit implementation. For this, run-of-mine samples were analysed using environmental scanning electron microscopy (ESEM) and infrared spectroscopy. The ESEM images showed that carbonate is present as cement and laminae. The infrared data revealed that the carbonate minerals correspond to aragonite and calcite, and that their occurrence is linked to the total amount of carbonate in the sample. By using a portable spectral instrument that uses diffuse reflectance, it was possible to classify the spectra of the ore samples based on the carbonate content. These results indicate that infrared technology could be used on-site for determining the quality of the ore, thus providing relevant information to assist the optimisation of mining and beneficiation activities. [ABSTRACT FROM AUTHOR]

The research was focused on the separation of single hemp (Cannabis sativa L.) fibre cells with low fineness from mechanically extracted fibre bundles of high fineness. The fiber bundles were treated with enzymes, namely panzym, pectinase, hemicellulase, and cellulase, along with a combination of panzym and ultrasonic treatments. Changes in the fiber structure were followed at molecular and microscopic levels by means of NIR FT Raman spectroscopy and Environmental Scanning Electron Microscopy (ESEM). Buffer-panzym treatments of hemp fibers had a prominent effect in loosening of the fiber cells. The best of refining was achieved when the fiber bundles were treated with buffer-panzym solution in combination with ultrasonic treatment.

Treatment of wood with aqueous dispersions of silica that have been modified with aluminum oxychloride (AlOCl) can impart wood water repellence and increased resistance to fungal decay. This study is a comparative survey on the effects of treatment with modified and non-modified silica dispersions in terms of the combustion behavior of the Scots pine (Pinus sylvestris L.) to evaluate the fire risk of this wood utilized as a construction material. The thermogravimetric results showed that treatments with the silica dispersions did not change the pyrolysis temperature of wood polymers, i.e. there was no synergetic interaction between silica and cell wall polymers during pyrolysis. Cone calorimetry (CONE) indicated that the silica-treated wood required longer time for ignition than the untreated control, but wood chars were similar. Treatments with both silica dispersions led to considerable reduction in the heat release (HR) and smoke production, but the cationically modified silica was more efficient than the unmodified silica. These findings were interpreted that incorporation of modified silica did not substantially influence the pyrolysis of cell wall polymers because they have not penetrated the cell wall; they have rather reduced the fire risk via forming a barrier against oxygen access and a thermal protection shield. [ABSTRACT FROM AUTHOR]

Environmental scanning electron microscopy (ESEM) permits to analyze samples in their native-hydrated state, allowing a broad spectrum of biological applications. In this study, ESEM equipped with energy dispersive X-ray spectrometer (EDS) was used as a fast method to analyze tissue morphology and to investigate metal distribution in the Ni hyperaccumulator Noccaea caerulescens , an established model to study the adaptation of plants to metalliferous soils. The low vacuum and wet mode operative conditions required the proper choice of experimental parameters both for morphological and compositional characterization of plant tissues. The calibration strategy for semi-quantitative analysis involved the use of Ni fortified agar as standard and signal normalization respect to endogenous carbon, chosen as internal standard. The obtained results are in accordance with present literature, showing a preferential Ni distribution in the epidermal cells respect to near the stomata for leaves and in the cotyledon epidermidis respect to cotyledon parenchyma area for seeds. Thanks to the absence of any time consuming sample treatment steps, ESEM-EDS technique can be proposed as valid strategy for in vivo high-throughput analysis of plant tissues and for a rapid screening and identification of other hyperaccumulator plants in a selected contaminated area. [ABSTRACT FROM AUTHOR]

In latex products manufacturing, protein has always been a problem due to the prevalence of latex protein allergy. Thus deproteinisation has been introduced to eliminate or reduce rubber latex protein content. However, the investigation on particles size and arrangement of particles in the rubber latex state before and after deproteinisation process has not been thoroughly explored. In this study, the particles size, arrangement and morphology of the film formed from modified latexes were monitored using Environmental Scanning Electron Microscopy (ESEM) and Field Emission Scanning Electron Microscopy (FESEM) for various latexes treated with either urea or sodium dodecyl sulphate and combination of both as deproteinisation agent. The size of latex particles changed after deproteinisation and the arrangement of particles when films are formed revealed interesting packing occurrence between the deproteinised latexes. These occurrences were shown to be associated to chemicals or surfactant used in the deproteinisation. The use of urea in combination with sodium dodecyl sulphate in the deproteinisation contributed to good latex particles packing during film formation attributed to the appearance of soft shell latex particles. Clearly, the latex particles and film morphologies is profoundly influenced by the type of deproteinisation agents used. [ABSTRACT FROM AUTHOR]

The current work investigates the redox behaviour of peak performing Ni–BaZr 0.85 Y 0.15 O 3−δ (Ni-BZY) cermet anodes for protonic ceramic fuel cells (PCFCs) by electrochemical impedance measurements, scanning electron microscopy (SEM) and X-ray diffraction (XRD). Peak performing PCFC cermet anodes are documented to require much lower porosity levels than those needed in oxide-ion conducting counterparts. The polarisation behaviour of these optimised PCFC anodes is shown to be drastically impaired by redox cycling, with depletions in performance that correspond to around 80% of the original resistance values noted after the first redox cycle. The ohmic resistance (Rohmic) is also shown to be increased due to delamination at the electrode/electrolyte interface, as confirmed by postmortem microstructural analysis. In-situ measurements by environmental scanning electron microscopy (ESEM) reveal that degradation proceeds due to volume expansion of the nickel phase during the re-oxidation stage of redox cycling. The present study reveals degradation to be very fast for peak performing Ni-BZY cermets of low porosity. Hence, methods to improve redox stability can be considered to be essential before such anodes can be implemented in practical devices. [ABSTRACT FROM AUTHOR]

Abstract: The use of cyclodextrins (CDs) for controlled delivery of drugs is largely presented in the literature. However, the question of whether CDs themselves linked to a polymeric network are able to sustain the release of drugs still persists. Here, CD immobilization within dextran microspheres is reported, and CD-dextran complexes were packed in a glass column and then, the retention time of different drugs and drug model compounds was determined by liquid chromatography. The release profiles of drugs and of drug model compounds (indole, 3-nitrophenol, p-hydroxybenzoic acid, diclofenac), characterized by different values of the retention time (high, moderate or low), were investigated. The release rates were quite high even for drugs that exhibit very high retention time (high association equilibrium constant). Moreover, the volume of the release fluid strongly influences the rate of drug release. As a whole, “the sink conditions” must be continuously maintained, since at each drug concentration in the release medium, equilibrium occurs between the free and the CD-bound drug. [Copyright &y& Elsevier]

Highlights: [•] Synthesis and characterization of nanowires. [•] Nanowires are unique 1-D nanostructures for chemical functionalization. [•] Nanowires for electrochemical sensing and biosensing. [•] Nanowires and electrochemical microfluidics. [•] Nanowires have plenty of possibilities on the horizon. [Copyright &y& Elsevier]

Abstract: This work examines the effect of the particle size of durum wheat semolina on the water diffusion coefficient and the evolution of the microstructure during hydration. Durum wheat semolina was sieved in 5 size categories. Water–particle interaction of powders (5 fractions and raw semolina) was studied using dynamic vapour sorption. Water sorption isotherms were fitted to G.A.B.. and Y&N models. Monolayer values as well as multilayer properties were calculated and compared. Absorption properties were determined by Y&N model. A sorption curve was modelled with the Fickian diffusion equation adapted to water diffusion into solid particles. A dynamic Fickian water diffusion coefficient was calculated for each particle size after conditioning at different RHs. The surface structure of conditioned fractions was observed by ESEM. The microstructure of semolina presented an evolution with the increase of size and RH. Starch grains presented an increased size and protein matrix had a different texture. After correlation of isotherm data and microstructure, the resulting state and properties of the components were a key to understand diffusion on food powder systems. [Copyright &y& Elsevier]

Abstract: Poly(N-isopropylacrylamide-co-N-hydroxyethylacrylamide) (poly(NIPAAm-co-HEAAm)) was prepared as a new thermosensitive copolymer possessing a sharp phase transition around the human body temperature. The effect of the copolymer concentration on the lower critical solution temperature (LCST) was determined under physiological conditions by cloud point (CP) and differential scanning calorimetric (DSC) methods. Then, thermosensitive microspheres were prepared from preformed copolymers by chemical cross-linking of hydroxyl groups with glutaraldehyde at a temperature situated slightly below LCST of the copolymer solution. The volume phase transition temperature (VPTT) of corresponding cross-linked microspheres was determined from swelling degree-temperature curve. The microspheres were loaded with model drug indomethacin by the solvent evaporation method. The DSC analysis proved that the drug is molecularly dispersed in the polymer network. Finally, the influence of the microsphere size on drug release was investigated. It was established that microspheres with the diameter ranging between 5 and 60μm release the drug with almost the same rate below (in the swollen state) and above the VPTT (in the collapsed state). On the contrary, microspheres with the diameter ranging between 125 and 220μm release a significantly higher amount of indomethacin below than above the VPTT. This different behavior is enough to assure a pulsatile release mechanism when the temperature changes cyclically below and above the VPTT. However, both small and large microspheres release a large amount of the drug during the collapsing process. [Copyright &y& Elsevier]

Highlights: [•] The EDX technique correctly predicts the surface composition of pure milk components powders and of model milk powders. [•] The technique was applied to whole and skim milk powders but differences in composition between surface and core were not observed. [•] The EDX technique coupled with XPS was successfully applied to obtain information on composition gradients between particle surface and core. [ABSTRACT FROM AUTHOR]

Abstract: This study developed a novel oral insulin formulation centered on microspheres consisting of a blend of biodegradable poly(ester amide) (PEA). In the formulation, l-lysine-/l-leucine-based PEA with pendant COOH groups (PEA-COOH) was used as a pH-responsive material for the protection of insulin from the harsh environmental conditions of the stomach. Arginine-based PEA (Arg-PEA) was introduced to improve the intestinal absorption of the drug. The influence of both the hydrophobicity of PEA-COOH and the content of Arg-PEA was investigated in detail on microsphere surface morphology, drug loading, and the in vitro release profile of insulin. The PEA-COOH/Arg-PEA blend microspheres protected the loaded insulin in simulated gastric fluid and released insulin in a fast and sustained manner in simulated intestinal fluid. The in vivo test demonstrated that the oral administration of insulin-loaded PEA blend microspheres could effectively suppress the blood glucose level in diabetic rats for 10h, and the oral bioavailability was improved to 5.89+1.84% in healthy rats. These results indicate that the PEA blend microspheres are promising vehicles for the oral delivery of insulin. [Copyright &y& Elsevier]

Abstract: New hyaluronic acid (HA)–itaconic acid (IT) films have been previously synthesized and used as potential topical drug delivery systems (DDS) for ocular administration. In this study we explored homogeneous and heterogeneous crosslinking reactions of HA using glutaraldehyde (GTA) and polyethylene glycol diglycidyl ether (PEGDE) in the presence of IT, a naturally occurring compound that is non-toxic and readily biodegradable. We have studied the morphology, mechanical properties and in vitro biocompatibility between these new materials and ocular surface cells (human corneal epithelial cell line) and evaluated the biopharmaceutical performance of the designed formulations. Although all the synthesized materials exhibited good mechanical properties, the PEGDE modified films exhibited the best biocompatibility, with in vivo assays showing good adhesive performance and minimal irritation. PEGDE films were also tested for their effects in the treatment of intraocular pressure (IOP) in rabbits using timolol maleate (TM) as the model drug. These results may be useful for further design of novel bioadhesive matrix containing drugs by topical application in ophthalmology. [Copyright &y& Elsevier]

This study explores the interaction between crack initiation and nanomechanical properties in the crack-tip fracture process zone of Carrara marble. Specimens with preexisting cracks were loaded in a uniaxial testing machine until the process zone appeared at the tips of the preexisting cracks. ESEM analysis reveals an increase in microcrack density in the process zone with increased loading of the specimen. Nanoindentation testing comprised of lines and grids of single nanoindentations located both near and far from the process zone shows a decrease in both indentation modulus and indentation hardness near grain boundaries in intact material, and with closeness to the process zone. Ultimately, the study confirms that the crack-tip process zone manifests itself as an area of reduced indentation hardness and indentation modulus in marble. [ABSTRACT FROM AUTHOR]

Laboratory water uptake tests are performed at the Belgian Nuclear Research Centre SCK•CEN to obtain insight into the hydromechanical behavior of Eurobitum bituminized radioactive waste under geological disposal conditions. Small nonradioactive and radioactive Eurobitum samples are hydrated in restricted swelling conditions (i.e., nearly constant volume conditions and constant stress conditions). Microfocus X-ray computer tomography (μCT) proves to be a very suitable technique to follow up the ingress of water in the samples. μCT analyses demonstrate that, under the studied hydration conditions, the water uptake by Eurobitum samples is a diffusion controlled process. A characterization of the partially leached samples with environmental scanning electron microscopy (ESEM) shows that the hydration of salt crystals and the subsequent dilution of the salt solution result in an increase in pore size that is limited to a few tens of μm in restricted swelling conditions. The μCT and ESEM analyses allow improvement in the understanding of water uptake by Eurobitum in restricted swelling conditions. In this article we discuss the μCT and ESEM analyses of nonradioactive Eurobitum samples that were hydrated for 2 to 4 years at a constant stress of 1, 22, 33, and 44 bar or in nearly constant volume conditions. [ABSTRACT FROM PUBLISHER]

Abstract: The potential application of a hydrogel-based strong anion-exchange (Q) membrane to purify plasmid DNAs was evaluated. The maximum binding capacity of plasmid DNA was estimated to be 12.4mg/ml of membrane volume with a plasmid recovery yield of ∼90%. The effect of the inherent properties of plasmid DNA, membrane adsorbent, and the ionic environment on membrane performance was systematically investigated. Plasmid DNAs with smaller tertiary structure tended to have a better recovery than those with larger tertiary structure. Environmental Scanning Electron Microscopy (ESEM) revealed that the hydrogel structure is more porous on one side of membrane than the other. Membrane pre-treatment significantly improved pore distribution and increased membrane porosity resulting in a better adsorption, recovery, and higher flux. The selection of proper operating pH led to further improvement. The relative contribution of these factors to improve membrane chromatography of plasmid DNAs was analyzed using statistical modeling. It was found that the adsorption of plasmid DNA was mainly affected by the available adsorptive area associated with membrane porosity, whereas the recovery of plasmid DNAs was mainly affected by the environmental pH. [Copyright &y& Elsevier]

Environmental scanning electron microscopy (ESEM) enables the investigation of hydrated and uncoated plant samples and the in situ observation of dynamic processes. Water vapor in the microscope chamber takes part in secondary electron detection and charge prevention. Two ESEM modes are available and offer a broad spectrum of applications. The environmental or wet mode prevents sample dehydration by the combination of sample cooling (5°C) and a vapor pressure of 4-6 Torr. In the low vacuum mode, the maximum chamber pressure is limited to 1 Torr (corresponding to about 5% relative humidity in the chamber) and allows the simultaneous use of a backscattered electron detector for imaging material contrast. A selection of characteristic plant samples and various applications are presented as a guide to ESEM for plant scientists. Leaf surfaces, trichomes, epicuticular waxes, and inorganic surface layers represent samples being comparatively resistant to dehydration, whereas callus cells and stigmatic tissue are examples for dehydration- and beam-sensitive samples. The potential of investigating dynamic processes in situ is demonstrated by studying anther opening, by tensile testing of leaves, and by performing hydration/dehydration experiments by changing the vapor pressure. Additionally, automated block-face imaging and serial sectioning using in situ ultramicrotomy is presented. The strengths and weaknesses of ESEM are discussed and it is shown that ESEM is a versatile tool in plant science. [ABSTRACT FROM AUTHOR]

In situ high-temperature healing of cracks in composites made of glass and vanadium boride (VB) particles was observed using an environmental scanning electron microscope equipped with a high-temperature chamber (HT-ESEM). HT-ESEM is an adequate tool for studying the self-healing property of these materials. The change in crack length as a function of redox atmospheric conditions is reported. No self-healing behaviour was observed under reducing conditions, while a complete and rapid healing of the cracks was measured under oxidizing conditions. HT-ESEM image analyses enabled the monitoring of the healing effect. The self-healing mechanism was identified as a consequence of the VB active particles oxidation and subsequent pouring of fluid oxides into the cracks. These innovative composites offer an interesting potential in the domain of solid oxide fuel cell sealants. [ABSTRACT FROM AUTHOR]

Abstract: Introduction: ProRoot mineral trioxide aggregate (MTA) and calcium silicate cements are able to set in a moist environment. The aim of the study was to examine the surface structure and composition of a cement paste under wet conditions and in real time during setting by environmental scanning electron microscopy connected with energy dispersive x-ray analysis (ESEM-EDX) and micro-Raman techniques. Methods: White ProRoot MTA and experimental white tetrasilicate cement (wTC) and wTC containing bismuth oxide (wTC-Bi) were studied. Cement disks were analyzed 10 minutes after powder-liquid mixing (freshly prepared samples) and after immersion in Dulbecco phosphate-buffered saline at 37°C for 24 hours (24-hour-aged samples). Results: Freshly prepared wet cements at ESEM-EDX exposed an irregular surface (displaying calcium, silicon, aluminum, chlorine reflexes, and bismuth traces in MTA and wTC-Bi) with needle-like and cubic-hexagonal shaped crystals. Aggregates of spheroidal Ca-P–rich crystals (spherulites) appeared on the surface of 24-hour-aged samples. The starting unhydrated powders displayed the typical Raman bands of Portland cement components: alite, belite, and calcium sulfate (only as anhydrite in MTA and as both anhydrite and gypsum in wTC and wTC-Bi). MTA powder showed higher amount of calcium carbonate and lower quantities of anhydrite and higher crystallinity of the silicate component, leading to a slower hydration reaction. Products/markers of hydration reactions were present on fresh samples; ettringite formed on the surface of all the cements; calcium hydroxide (portlandite) was detected only on the surface of wTC, but no conclusion can be drawn on wTC-Bi and MTA because of the interference of bismuth oxide. Calcium phosphate and calcite/aragonite bands were detected on all 24-hour-aged cements; portlandite was no longer detected on wTC. Conclusions: ESEM and micro-Raman are powerful and suitable techniques to investigate endodontic calcium silicate hydrated cements in real time and in their humid state without inducing artifacts by sample preparation. The formation of apatite spherulites on calcium silicate cements might have clinical relevance. [Copyright &y& Elsevier]

Abstract: Groundwater and core sediments of two boreholes (to a depth of 50m) from the Chapai-Nawabganj area in northwestern Bangladesh were collected for arsenic concentration and geochemical analysis. Groundwater arsenic concentrations in the uppermost aquifer (10–40m of depth) range from 2.8μgL−1 to 462.3μgL−1. Groundwater geochemical conditions change from oxidized to successively more reduced, higher As concentration with depth. Higher sediment arsenic levels (55mgkg−1) were found within the upper 40m of the drilled core samples. X-ray absorption near-edge structure spectroscopy was employed to elucidate the arsenic speciation of sediments collected from two boreholes. Environmental scanning electron microscopy and transmission X-ray microscopy were used to investigate the characteristics of FeOOH in sediments which adsorb arsenic. In addition, a pH–Eh diagram was drawn using the Geochemist''s Workbench (GWB) software to elucidate the arsenic speciation in groundwater. The dominant groundwater type is Ca–HCO3 with high concentrations of As, Fe and Mn but low levels of NO3 − and SO4 2−. Sequential extraction analysis reveals that Mn and Fe hydroxides and organic matter are the major leachable solids carrying As. High levels of arsenic concentration in aquifers are associated with fine-grained sediments. Fluorescent intensities of humic substances indicate that both groundwater and sediments in this arsenic hotspot area contain less organic matter compared to other parts of Bengal basin. Statistical analysis clearly shows that As is closely associated with Fe and Mn in sediments while As is better correlated with Mn in groundwater. These correlations along with results of sequential leaching experiments suggest that reductive dissolution of MnOOH and FeOOH mediated by anaerobic bacteria represents an important mechanism for releasing arsenic into the groundwater. [Copyright &y& Elsevier]

Three water soluble surfactants based on nonyl phenol were prepared, namely: ethoxylated nonyl phenol (e.o = 9), E(9)NP, ethoxylated nonyl phenol sodium phosphate, E(9)NPSP, and ethoxylated nonyl phenol phosphate diamide, E(9)NPPA. The inhibition efficiency of these surfactants on H-11 carbon steel in 1 M HCL was investigated using the weight loss technique. The surface active and thermodynamic properties of the investigated inhibitors have been studied. The surface morphology of carbon steel in absence and presence of different concentrations of E(9)NPPA was examined by environmental scanning electron microscopy (ESEM). The maximum inhibition efficiency (93.21%) was obtained by E(9)NPPA at 300 ppm and 298°K. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this work was to study the behaviour of zinc-doped phosphosilicate glasses based on Bioglass® 45S5. In vitro (in simulated body fluid), the reactivity was analysed by means of inductively coupled plasma spectrometry, environmental scanning electron microscopy–energy-dispersive spectroscopy (ESEM–EDS) and X-ray diffraction. In vivo (a rat implanted with glass), the reactivity and the tissue behaviour were analysed by conventional histology, histochemistry, microradiography and ESEM–EDS. The in vivo behaviour matches that in vitro perfectly; they show comparable glass degradation processes and rates, ruled by the amount of zinc in the glass. The reaction mechanism for the formation of a polymerized silica layer superimposed with a peripheral calcium phosphate layer is clearly substantiated by ESEM–EDS investigations. The crystallization of a biologically active hydroxyapatite (HA) layer is observed in both cases; the in vitro experiment shows the presence of HA after 4 days. [Copyright &y& Elsevier]

Abstract: Fine particulate matter with aerodynamic diameter less than 2.5μm (PM2.5) was collected at urban and suburban sites in Shanghai from April 2005 to March 2006. Average mass concentrations of PM2.5 ranged from 43.5 to 149μgm−3 in the urban area and 21.7 to 159μgm−3 in suburban area. The mass levels of PM2.5 sampled at urban and suburban sites showed seasonal variation with much higher values in winter and spring, lower values in summer, and the lowest in autumn. The results of environmental scanning electron microscopy (ESEM) showed that Shanghai PM2.5 was consisted of soot aggregates, coal fly ashes, minerals, bio-particles and unidentified particles. Inductively coupled plasma atomic mission spectrum (ICP-AES) results showed total elements in Shanghai PM2.5 increased gradually from summer to winter and remained at a relatively high level in spring. There was a significant difference in the mass of elements in PM2.5 collected in urban and in suburban atmosphere. Enrichment factor (EF) analysis results demonstrated that K, Na, Ca, Mg, Al, Fe, Ba and Sr originated from natural sources, while As, Cu, Zn, Pb, Cd, Mn, Ni and Se were emitted from anthropogenic sources. The plasmid DNA assay showed that potential toxicity of Shanghai PM2.5 collected at urban and suburban sampling sites, and in different seasons, varied greatly. Toxicity of the Shanghai urban winter PM2.5 sample was much stronger compared to any of the other samples. Heavy metals in Shanghai PM2.5, including Cu, Zn, Pb, Cd, Cr, Mn, and Ni, might have synergic-effects on plasmid DNA damage. [Copyright &y& Elsevier]

Abstract: Activated carbon adsorbents were produced from biomass locally available in the Rocky Mountain West, e.g. Lodgepole Pine (Pinus contorta), by vacuum pyrolysis at moderate temperatures followed by steam activation, for use as metal adsorbents for acid mine drainage (AMD). Wood cubes from fresh cut Lodgepole Pine (P. contorta) with different sizes, 3 and 12mm, were made. Sawdust was also used to study the effect of sample size as well as sample material. We applied chemical pretreatment with potassium hydroxide before charring to improve the quality of the activated carbons. We compared the characteristics of the activated carbons, which were chemically pretreated, before and after washing with water. After washing, the BET surface area was found to increase and diffuse reflectance infrared spectroscopy showed changes in the carbon matrix. We then tested the samples for metal adsorption from AMD sampled from AMD sites in Colorado, Clear Creek County and the Leadville mine drainage tunnel, along with a commercial activated carbon for comparison. We used a batch method to measure maximum metal adsorption of the activated carbons. The metals chosen to be monitored were copper, cadmium, manganese, nickel, lead, and zinc, because they are the principal metals of interest for the test areas, and metal concentrations were determined by ion coupled plasma–atomic emission spectroscopy. The samples produced in this work outperformed the commercial activated carbon in two AMD water treatment tests and for the six metals monitored. This metal adsorption data indicate that locally produced inexpensive activated carbons can be used as adsorbents for AMD successfully. [Copyright &y& Elsevier]

Abstract: In this study we have investigated the chemical–physical and morphological properties of intact and atelocollagen sponges used for tissue engineering. The porous sponges were prepared by lyophilization and their physico-chemical characteristics (water binding capacity, denaturing temperature, amino group content) were investigated. Considering the importance of the “in vivo” interactions between these sponges and the tissue, our attention was addressed (a) to clarify the relationships between the morphology and the amount of water absorbed and (b) to evaluate the influence of pepsin-alkaline treatment on the reorganization of the atelocollagen fibres. Conventional scanning electron microscopy (SEM) and environmental scanning electron microscopy (ESEM) were employed to study the morphology and wetting behaviour of the intact and atelocollagen sponges. The observations by SEM indicated remarkable differences both in the structure and dimension of the pores between intact and atelocollagen sponges. At the data are related to a different water binding capacity. However, the ESEM observations, achieved by changing the relative humidity in the operative chamber, demonstrated that the water adsorbed can be removed with major difficulty from atelocollagen sponges than from intact ones. [Copyright &y& Elsevier]

Abstract: The mechanism of the reaction between a silica sample coming from acid treatment of sepiolite (denominated Silsep) and an organosilane, namely bis(triethoxysilylpropyl)tetrasulfane (TESPT), has been evaluated by solid state NMR spectroscopy, being compared with the silanization reaction of a commercial silica. The effect of the silane concentration and temperature on the course of the reaction was considered. Experimental results indicate that the silanization reaction is more effective in the case of Silsep, favoring both the reaction of silane molecules with the filler surface and the reaction between neighboring silane molecules. This different behavior is attributed to structural factors, moisture, and number of acid centers on silica surface. Environmental scanning electron microscopy (ESEM) was used to deposit micrometric water drops on the surface of these samples and to evaluate the proportion and distribution of the organophylization process. [Copyright &y& Elsevier]

Abstract: Prosthetic meshes are used as the standard of care in abdominal wall hernia repair. However, hernia recurrences and side effects remain unsolved problems. The demand by health care providers for increasingly efficient and cost-effective surgery encourages the development of newer strategies to improve devices and outcomes. Here, we evaluated whether l-arginine administration was able to ameliorate long-term polypropylene prostheses incorporation into the abdominal wall of Sprague–Dawley rats. Meshes were placed on-lay and continuous l-arginine was administered. In vivo biocompatibility was studied at 7, 25 and 30 days post-implantation. Effectively, l-arginine administration in combination with mesh triggered subtle changes in ECM composition that impinged on critical biochemical and structural features. Lastly, tensile strength augmented and stiffness decreased over the control condition. This could help to restructure the mechanical load transfer from the implant to the brittle surrounding tissues, i.e., impact load and fatigue load associated with mechanical tensions could be distributed between the mesh and the restored tissue in a more balanced manner, and ultimately help to reduce the incidence of loosening, recurrences, and local wound complications. Since the newly formed tissue is more mechanically stable, this approach could eventually be introduced to human hernia repair. [Copyright &y& Elsevier]

Characterization of protein-fouled membranes is crucial to understand the fouling mechanisms and optimization of fouling control and cleaning. This paper reviews current developments in membrane characterization as applied to protein-fouled membranes and difficulties still encountered. The potential of a number of new techniques yet to be fully exploited are also discussed. [Copyright &y& Elsevier]

Two potash-lime-silica model glasses with compositions similar to those of medieval stained glass were exposed at 26 test sites all over Europe and in North America for 3–6 years. The objectives of this large-scale field exposure programme were: (a) a qualitative analysis of the weathering products formed on the surface of the samples in the (environmental) scanning electron microscope in combination with energy dispersive X-ray microanalysis (ESEM/EDX or SEM/EDX, respectively); (b) a statistical evaluation of the weathering phenomena by applying a multiple linear regression (MLR) analysis to find correlations between the degree of weathering, which was measured in terms of surface coverage with reaction products, and environmental parameters such as the concentration (c) of the acidifying gases SO2, NO2 and O3, the temperature (T) and the relative humidity (RH); and (c) the calculation of a time-dependence function of the weathering process of these glasses. Mainly sulfates of calcium and potassium such as gypsum (CaSO4·2 H2O), arcanite (K2SO4) and syngenite (CaSO4·K2SO4·H2O) could be identified in the SEM and ESEM. Carbonates, nitrates and many particles deposited on the glass surface were found as well. MLR calculations exhibit significant dependencies of the degree of weathering on T, RH, c(NOx), c(SO2) and c(O3). Applying a time-dependence function of the general form y=a·tb (t=time) results in a value of approximately 0.42 for the exponent b, which comes close to values expected from various studies in the literature. [ABSTRACT FROM AUTHOR]

The application of a micro-sphere plate (MSP) to secondary electron detection in environmental scanning electron microscopy (ESEM) and possible ways of using the MSP is discussed and experimental measurements made of significant MSP parameters. In the preferred solution the MSP plays the role of a semi-permeable wall separating a standard scintillator detector from the low vacuum chamber. The gas conductance measured for an MSP of dimensions 1.4 mm thick and 12 mm diameter was 0.03 dm3/s, which is sufficiently low and suitable for this application. A survey of the MSP multiplication coefficient for various gases and vapours revealed that the coefficient increases at increasing gas pressures and has a value of the order of 102–103 in the pressure range 10−2–10−1 mbar. The results suggest that the MSP may be suitable as a secondary electron detector for ESEM. [Copyright &y& Elsevier]

Summary • The association of plants with arbuscular mycorrhizal (AM) fungi is widespread in nature, but little is known about molecular aspects of this symbiosis. Particularly during the early stages of the AM symbiosis, it is difficult to monitor growth of the two partners, to dissect gene expression patterns and to correlate them with plant, fungal or symbiosis development. • A new system, the ‘mini-mycorrhizotron’, was established to cultivate seedlings of Medicago truncatula in mycorrhizal symbiosis with Glomus intraradices under gnotobiotic conditions. This system allows natural growth of the symbiotic partners and permits the continuous noninvasive observation of the development of plant and fungus under a microscope. • The mini-mycorrhizotron was used to determine the stage of induction of a mycorrhiza-related gene detected by differential display-reverse transcription-PCR, namely a novel chalcone synthase (Mt-chs1). The gene is induced in roots at the stage of the first fungal contact. • The mini-mycorrhizotron allowed identification and cloning of a symbiosis-related gene, and the correlation between its expression and the developmental stage of the symbiosis was established. This provides a useful tool for molecular and developmental studies of the early stages of AM symbioses. [ABSTRACT FROM AUTHOR]

[Display omitted] Human respiratory mucus lining the airway epithelium forms a challenging barrier to inhalation therapeutics. Therefore, structural elucidation of hydrated mucus is essential for an efficient drug delivery development. The structure of mucus has been primarily investigated by conventional electron microscopy techniques, which operate under vacuum conditions and require sample preparation steps that might alter the structure of mucus. In this study we investigated the impact of dehydration on mucus and analyzed the structure of mucus in its hydrated state. Cryo-scanning electron microscopy (Cryo-SEM) analysis of mucus showed, that during the process of sublimation, non-porous structure of mucus is transformed into a porous network. Similarly, images acquired by environmental scanning electron microscopy (ESEM), revealed a non-porous structure of hydrated mucus, while further observation at decreasing pressure demonstrated the strong influence of dehydration on mucus structure. We could successfully visualize the structural organization of the major gel forming mucin MUC5B in its hydrated state by employing stimulated emission depletion (STED) microscopy, which allowed resolving the nano-scale patterns of mucin macromolecules within the essentially pore-free mucus structure. The general structural organization of mucus components was addressed by confocal laser scanning microscopy (CLSM), which revealed the heterogeneous and composite structure of mucus. These results provide a novel view on the native structure of mucus and will affect drug delivery development. [ABSTRACT FROM AUTHOR]