The influence of solvent refractive index on the photoluminescence decay of thick-shell gradient-alloyed colloidal quantum dots investigated in a wide range of delay times.

Colloidal semiconductor quantum dots have many potential optical applications, including quantum dot light-emitting diodes, single-photon sources, or biological luminescent markers. The optical properties of colloidal quantum dots can be affected by their dielectric environment. This study investigated the photoluminescence (PL) decay of thick-shell gradient-alloyed colloidal semiconductor quantum dots as a function of solvent refractive index. These measurements were conducted in a wide range of delay times to account for both the initial spontaneous decay of excitons and the delayed emission of excitons that has the form of a power law. It is shown that whereas the initial spontaneous PL decay is very sensitive to the refractive index of the solvent, the power-law delayed emission of excitons is not. Our results seem to exclude the possibility of carrier self-trapping in the considered solvents and suggest the existence of trap states inside the quantum dots. Finally, our data show that the average exciton lifetime significantly decreases as a function of the solvent refractive index. The change in exciton lifetime is qualitatively modeled and discussed.

Structural effects of charge destabilization and amino acid substitutions in amyloid fragments of CsgA.

The most studied functional amyloid is the CsgA, major curli subunit protein, which is produced by numerous strains of Enterobacteriaceae. Although CsgA sequences are highly conserved, they exhibit species diversity, which reflects the specific evolutionary and functional adaptability of the major curli subunit. Herein, we performed bioinformatics analyses to uncover the differences in the amyloidogenic properties of the R4 fragments in Escherichia coli and Salmonella enterica and proposed four mutants for more detailed studies: M1, M2, M3, and M4. The mutated sequences were characterized by various experimental techniques, such as circular dichroism, ATR-FTIR, FT-Raman, thioflavin T, transmission electron microscopy and confocal microscopy. Additionally, molecular dynamics simulations were performed to determine the role of buffer ions in the aggregation process. Our results demonstrated that the aggregation kinetics, fibril morphology, and overall structure of the peptide were significantly affected by the positions of charged amino acids within the repeat sequences of CsgA. Notably, substituting glycine with lysine resulted in the formation of distinctive spherically packed globular aggregates. The differences in morphology observed are attributed to the influence of phosphate ions, which disrupt the local electrostatic interaction network of the polypeptide chains. This study provides knowledge on the preferential formation of amyloid fibrils based on charge states within the polypeptide chain.

Detailed Insight into Photocatalytic Inactivation of Pathogenic Bacteria in the Presence of Visible-Light-Active Multicomponent Photocatalysts.

The use of heterogeneous photocatalysis in biologically contaminated water purification processes still requires the development of materials active in visible light, preferably in the form of thin films. Herein, we report nanotube structures made of TiO2/Ag2O/Au0, TiO2/Ag2O/PtOx, TiO2/Cu2O/Au0, and TiO2/Cu2O/PtOx obtained via one-step anodic oxidation of the titanium-based alloys (Ti94Ag5Au1, Ti94Cu5Pt1, Ti94Cu5Au1, and Ti94Ag5Pt1) possessing high visible light activity in the inactivation process of methicillin-susceptible S. aureus and other pathogenic bacteria-E. coli, Clostridium sp., and K. oxytoca. In the samples made from Ti-based alloys, metal/metal oxide nanoparticles were formed, which were located on the surface and inside the walls of the NTs. The obtained results showed that oxygen species produced at the surface of irradiated photocatalysts and the presence of copper and silver species in the photoactive layers both contributed to the inactivation of bacteria. Photocatalytic inactivation of E. coli, S. aureus, and Clostridium sp. was confirmed via TEM imaging of bacterium cell destruction and the detection of CO2 as a result of bacteria cell mineralization for the most active sample. These results suggest that the membrane ruptures as a result of the attack of active oxygen species, and then, both the membrane and the contents are mineralized to CO2.

Single Gold Nanobipyramids Sensing the Chirality of Amyloids.

Plasmonic nanoparticles, due to their sensitivity to small changes in their closest environment and plasmon resonance, can sense the chirality of the surrounding molecules. Therefore, plasmonic nanoparticles can be applied as a next-generation biosensor for peptides or proteins. In this work, we explore the interaction between chiral, ordered protein aggregates (amyloids) and small gold nanobipyramids. We show how the morphology, structure, and chiroptical properties of amyloids induce circular dichroism in the plasmon resonance wavelengths from individual plasmonic nanoparticles upon binding to the chiral amyloid template. Moreover, using the data from microscopic and spectroscopic analyses of formed heterostructures, we propose the most probable mechanism behind the induction of chirality in this system and discuss which specific feature of insulin protein aggregates is sensed by nanobipyramids.

Chitin scaffolds derived from the marine demosponge Aplysina fistularis stimulate the differentiation of dental pulp stem cells.

The use of stem cells for tissue regeneration is a prominent trend in regenerative medicine and tissue engineering. In particular, dental pulp stem cells (DPSCs) have garnered considerable attention. When exposed to specific conditions, DPSCs have the ability to differentiate into osteoblasts and odontoblasts. Scaffolds are critical for cell differentiation because they replicate the 3D microenvironment of the niche and enhance cell adhesion, migration, and differentiation. The purpose of this study is to present the biological responses of human DPSCs to a purified 3D chitin scaffold derived from the marine demosponge Aplysina fistularis and modified with hydroxyapatite (HAp). Responses examined included proliferation, adhesion, and differentiation. The control culture consisted of the human osteoblast cell line, hFOB 1.19. Electron microscopy was used to examine the ultrastructure of the cells (transmission electron microscopy) and the surface of the scaffold (scanning electron microscopy). Cell adhesion to the scaffolds was determined by neutral red and crystal violet staining methods. An alkaline phosphatase (ALP) assay was used for assessing osteoblast/odontoblast differentiation. We evaluated the expression of osteogenic marker genes by performing ddPCR for ALP, RUNX2, and SPP1 mRNA expression levels. The results show that the chitin biomaterial provides a favorable environment for DPSC and hFOB 1.19 cell adhesion and supports both cell proliferation and differentiation. The chitin scaffold, especially with HAp modification, isolated from A. fistularis can make a significant contribution to tissue engineering and regenerative medicine.

The application of the hierarchical approach for the construction of foldameric peptide self-assembled nanostructures.

In this paper, we show that a hierarchical approach for the construction of nanofibrils based on α,ß-peptide foldamers is a rational method for the design of novel self-assembled nanomaterials based on peptides. Incorporation of a trans-(1S,2S)-2-aminocyclopentanecarboxylic acid residue into the outer positions of the model coiled-coil peptide led to the formation of helical foldamers, which was determined by circular dichroism (CD) and vibrational spectroscopy. The oligomerization state of the obtained peptides in water was established by analytical ultracentrifugation (AUC). The thioflavin T assay and Congo red methods showed that the obtained α,ß-peptides possess a strong tendency to aggregate, leading to the formation of self-assembled nanostructures, which were assessed by microscopic techniques. The location of the ß-amino acid in the heptad repeat of the coiled-coil structure proved to have an influence on the secondary structure of the obtained peptides and on the morphology of the self-assembled nanostructures.

Photocatalytic hydrogen evolution from glycerol-water mixture under visible light over zinc indium sulfide (ZnIn2S4) nanosheets grown on bismuth oxychloride (BiOCl) microplates.

ZnIn2S4 (ZIS) is one of the widely studied photocatalyst for photocatalytic hydrogen evolution applications due to its prominent visible light response and strong reduction ability. However, its photocatalytic glycerol reforming performance for hydrogen evolution has never been reported. Herein, the visible light driven BiOCl@ZnIn2S4 (BiOCl@ZIS) composite was synthesized by growth of ZIS nanosheets on a template-like hydrothermally pre-prepared wide-band-gap BiOCl microplates using simple oil-bath method to be used for the first time for photocatalytic glycerol reforming for photocatalytic hydrogen evolution (PHE) under visible light irradiation (λ > 420 nm). The optimum amount of BiOCl microplates in the composite was found 4 wt% (4% BiOCl@ZIS) in the presence of in-situ 1 wt% Pt deposition. Then, the in-situ Pt photodeposition optimization studies over 4% BiOCl@ZIS composite showed the highest PHE rate of 674 µmol g-1h-1 with the ultra-low platinum amount (0.0625 wt%). The possible mechanisms behind this improvement can be ascribed to the formation of Bi2S3 low-band-gap semiconductor during BiOCl@ZIS composite synthesis resulting in Z-scheme charge transfer mechanism between ZIS and Bi2S3 upon visible light irradiation. This work expresses not only the photocatalytic glycerol reforming over ZIS photocatalyst but also a solid proof of the contribution of wide-band-gap BiOCl photocatalysts to enhancement of ZIS PHE performance under visible light.

Light-Induced In Situ Transmission Electron Microscopy─Development, Challenges, and Perspectives.

Transmission electron microscopy is a basic technique used for examining matter at the highest magnification scale available. One of its most challenging branches is in situ microscopy, in which dynamic processes are observed in real time. Among the various stimuli, like strain, temperature, and magnetic or electric fields, the light-matter interaction is rarely observed. However, in recent years, a significant increase in the interest in this technique has been observed. Therefore, I present a summary and critical review of all the in situ experiments performed with light, various technical possibilities for bringing radiation inside the transmission electron microscope, and the most important differences between the effects of light and electrons on the studied matter. Finally, I summarize the most promising directions for further research using light excitation.

Affordable Open-Source Quartz Microbalance Platform for Measuring the Layer Thickness.

The layer thickness measurement process is an indispensable companion of vacuum sputtering and evaporation. Thus, quartz crystal microbalance is a well-known and reliable method for monitoring film thickness. However, most commercial devices use very simple signal processing methods, offering only a readout of the frequency change value and an approximate sputtering rate. Here, we show our concept of instrument, to better control the process parameters and for easy replication. The project uses open-source data and its own ideas, fulfilling all the requirements of a measuring system and contributing to the open-source movement due to the added value and the replacement of obsolete technologies with contemporary ones. The device provides an easy way to expand existing sputtering machines with a proper controller based on our work. The device described in the paper can be easily used in need, being a proven project of a fast, inexpensive, and reliable thin-film thickness monitor.

Light-induced In Situ Transmission Electron Microscopy for Observation of the Liquid-Soft Matter Interaction.

The current protocol describes the modifications of the transmission electron microscope (TEM) setup for in situ light-induced observations. A glass optical fiber inserted into the electron column above the objective lens polepiece, and a laser, an adjustable light source, was used to fabricate the device. After the illuminator has been calibrated using an external measuring system, it allows one to adjust the intensity of the lighting to the needs of the observed process. This lighting system was utilized to image antimicrobial photodynamic therapy phenomena, which are currently the subject of intense research. The sample was prepared by spotting a suspension of bacteria on a carbon, graphene, or silicon nitride substrate, blotting the excess solution, spotting the photosensitizer solution, blotting the excess liquid again, and then assembling the liquid cell with a second substrate or graphene film. The process of the imaging experiment itself includes choosing the right place for observation with the use of low magnification and a minimum dose of electrons, and then cyclical activation of the light source to capture subsequent images at specified intervals with the minimum amount of electrons necessary. The electron dose of each exposure and the time and intensity of lighting used need to be carefully recorded due to the complexity of the observed phenomena as, at the same time, the process is both light- and electron-driven. After the actual experiment is performed, additional control observations must be made, in which the same doses of electrons are used but without additional light influence and smaller doses of electrons are used for higher doses of light. This makes it possible to distinguish light-induced microstructural effects from those caused by electrons in both the fields of life and materials science.

Molecular Level Characterisation of the Surface of Carbohydrate-Functionalised Mesoporous silica Nanoparticles (MSN) as a Potential Targeted Drug Delivery System via High Resolution Magic Angle Spinning (HR-MAS) NMR Spectroscopy.

Atomistic level characterisation of external surface species of mesoporous silica nanoparticles (MSN) poses a significant analytical challenge due to the inherently low content of grafted ligands. This study proposes the use of HR-MAS NMR spectroscopy for a molecular level characterisation of the external surface of carbohydrate-functionalised nanoparticles. MSN differing in size (32 nm, 106 nm, 220 nm) were synthesised using the sol-gel method. The synthesised materials displayed narrow particle size distribution (based on DLS and TEM results) and a hexagonal arrangement of the pores with a diameter of ca. 3 nm as investigated with PXRD and N2 physisorption. The surface of the obtained nanoparticles was functionalised with galactose and lactose using reductive amination as confirmed by FTIR and NMR techniques. The functionalisation of the particles surface did not alter the pore architecture, structure or morphology of the materials as confirmed with TEM imaging. HR-MAS NMR spectroscopy was used for the first time to investigate the structure of the functionalised MSN suspended in D2O. Furthermore, lactose was successfully attached to the silica without breaking the glycosidic bond. The results demonstrate that HR-MAS NMR can provide detailed structural information on the organic functionalities attached at the external surface of MSN within short experimental times.

N'-terminal- and Ca2+-induced stabilization of high-order oligomers of full-length Danio rerio and Homo sapiens otolin-1.

Otolin-1 is a C1q family member and a major component of the organic matrix of fish otoliths and human otoconia. To date, the protein molecular properties have not been characterized. In this work, we describe biochemical characterization and comparative studies on saccular-specific otolin-1 derived from Danio rerio and Homo sapiens. Due to the low abundance of proteins in the otoconial matrix, we developed a production and purification method for both recombinant homologues of otolin-1. Danio rerio and Homo sapiens otolin-1 forms higher-order oligomers that can be partially disrupted under reducing conditions. The presence of Ca2+ stabilizes the oligomers and significantly increases the thermal stability of the proteins. Despite the high sequence coverage, the oligomerization of Danio rerio otolin-1 is more affected by the reducing conditions and presence of Ca2+ than the human homologue. The results show differences in molecular behaviour, which may be reflected in Danio rerio and Homo sapiens otolin-1 role in otolith and otoconia formation.

Amyloid fibrils in superstructures - local ordering revealed by polarization analysis of two-photon excited autofluorescence.

Protein misfolding products - amyloids - tend to form distinct fibrillar structures of the characteristic fold for a given neurodegenerative disease or pathology. Moreover, amyloids (also in the intermediate or distorted state) can act as secondary nuclei for de novo fibrillation. Such secondary nucleation amplifies plaque development correlated with various diseases. Therefore, a versatile and non-destructive method of detection and differentiation between distinct fibrillar structures is of great importance. Amyloids exhibit unique optical properties, i.e. green-blue autofluorescence, which can also be induced by two-photon excitation. Herein, we use this label-free technique to resolve local fibrillar ordering in amyloid superstructures - spherulites. With polarization-dependent two-photon excited amyloid autofluorescence, we resolved fibrillar orientation in the spherulite corona and discussed the presence of amorphous aggregates, distorted fibrils or amyloid intermediate species within the spherulite core. Our polarization sensitive two-photon microscopy investigations are supported by TEM imaging and provide a promising tool for the detection and differentiation between well-developed amyloid fibrils and amorphous/distorted structures present at different stages of the formation of amyloid superstructures and plaques.

A Method for Underwater Wireless Data Transmission in a Hydroacoustic Channel under NLOS Conditions.

Wireless data transmission in the hydroacoustic channel under non-line-of-sight (NLOS) propagation conditions, for example, during a wreck penetration, is difficult to implement reliably. This is mostly due to the multipath propagation, which causes a reduction in the quality of data reception. Therefore, in this work an attempt has been made to develop a reliable method of wireless underwater communication test it under the NLOS conditions. In our method, we used multiple frequency-shift keying (MFSK) modulation, sending a single bit on two carriers, and diversity combining. The method was tested in laboratory conditions which simulated underwater signal propagation during the penetration of the wreck. The propagation conditions were investigated by determining the impulse responses at selected measurement points using the correlation method. Additionally, for comparison, the data transmission quality was determined by the bit error rate (BER) under the same conditions using direct sequence spread spectrum (DSSS) and binary phase shift keying (BPSK) modulation. The obtained results confirmed the usefulness of the application of the developed method for wireless data transmission in a hydroacoustic channel under NLOS conditions.

Fiber-optic sample illuminator design for the observation of light induced phenomena with transmission electron microscopy in situ: Antimicrobial photodynamic therapy.

Antibacterial photodynamic therapy is a promising treatment for problematic infections caused by bacteria and fungi. Despite its undoubted effectiveness, the ultrastructural mechanism of microbial death remains not fully described and distinct organisms respond to the treatment with different efficacy. For this reason, it was decided to try imaging the process using the in situ transmission electron microscopy method. To conduct an observational experiment, the microscope was significantly modified. Liquid cell methods were used, electron doses and their influence on the sample were estimated, and a fiber-optic sample illuminator was designed and built. The modifications allowed for the light-induced characterization of photosensitizer-bacteria interaction. Microscope modification is a promising platform for further studies of light-induced phenomena in both life and material science.

Nucleobindin-2 consists of two structural components: The Zn2+-sensitive N-terminal half, consisting of nesfatin-1 and -2, and the Ca2+-sensitive C-terminal half, consisting of nesfatin-3.

Nucleobindin-2 (Nucb2) is a protein that has been suggested to play roles in a variety of biological processes. Nucb2 contains two Ca2+/Mg2+-binding EF-hand domains separated by an acidic amino acid residue-rich region and a leucine zipper. All of these domains are located within the C-terminal half of the protein. At the N-terminal half, Nucb2 also possesses a putative Zn2+-binding motif. In our recent studies, we observed that Nucb2 underwent Ca2+-dependent compaction and formed a mosaic-like structure consisting of intertwined disordered and ordered regions at its C-terminal half. The aim of this study was to investigate the impact of two other potential ligands: Mg2+, which possesses chemical properties similar to those of Ca2+, and Zn2+, for which a putative binding motif was identified. In this study, we demonstrated that the binding of Mg2+ led to oligomerization state changes with no significant secondary or tertiary structural alterations of Nucb2. In contrast, Zn2+ binding had a more pronounced effect on the structure of Nucb2, leading to the local destabilization of its N-terminal half while also inducing changes within its C-terminal half. These structural rearrangements resulted in the oligomerization and/or aggregation of Nucb2 molecules. Taken together, the results of our previous and current research help to elucidate the structure of the Nucb2, which can be divided into two parts: the Zn2+-sensitive N-terminal half (consisting of nesfatin-1 and -2) and the Ca2+-sensitive C-terminal half (consisting of nesfatin-3). These results may also help to open a new discussion regarding the diverse roles that metal cations play in regulating the structure of Nucb2 and the various physiological functions of this protein.

CdS Nanoplates Modification as a Platform for Synthesis of Blue-Emitting Nanoparticles.

In this paper, the study of surface modification of two-dimensional (2D), non-luminescent CdS nanoplates (NPLs) by thiol-containing ligands is presented. We show that a process of twophase transfers with appropriate ligand exchange transforms non-luminescent NPLs into spherical CdS nanoparticles (NPs) exhibiting a blue photoluminescence with exceptionally high quantum yield ~90%. In the process, transfer from inorganic solvent to water is performed, with appropriately selected ligand molecules and pH values (forward phase transfer), which produces NPs with modified size and shape. Then, in reverse phase transfer, NPs are transferred back to toluene due to surface modification by combined Cd (OL)2 and Cd (Ac)2. As a result, spherical NPs are formed (average diameter between 4 and 6 nm) with PL QY as high as 90%. This is unique for core only CdS NPs without inorganic shell.

Naturally Formed Chitinous Skeleton Isolated from the Marine Demosponge Aplysina fistularis as a 3D Scaffold for Tissue Engineering.

Tissue engineering (TE) is a field of regenerative medicine that has been experiencing a special boom in recent years. Among various materials used as components of 3D scaffolds, naturally formed chitinous materials seem to be especially attractive because of their abundance, non-toxic and eco-friendly character. In this study, chitinous skeleton isolated from the marine sponge Aplysina fistularis (phylum: Porifera) was used for the first time as a support for the cultivation of murine fibroblasts (Balb/3T3), human dermal fibroblasts (NHDF), human keratinocyte (HaCaT), and human neuronal (SH-SY5Y) cells. Characterization techniques such as ATR FTIR, TGA, and µCT, clearly indicate that an interconnected macro-porous, thermostable, pure α-chitin scaffold was obtained after alkali-acid treatment of air-dried marine sponge. The biocompatibility of the naturally formed chitin scaffolds was confirmed by cell attachment and proliferation determined by various microscopic methods (e.g., SEM, TEM, digital microscopy) and specific staining. Our observations show that fibroblasts and keratinocytes form clusters on scaffolds that resemble a skin structure, including the occurrence of desmosomes in keratinocyte cells. The results obtained here suggest that the chitinous scaffold from the marine sponge A. fistularis is a promising biomaterial for future research about tissues regeneration.

Light-induced in situ transmission electron microscopy: Novel approach for antimicrobial photodynamic therapy imaging.

The novel approach for imaging of antimicrobial photodynamic therapy processes presented in this work is based on transmission electron microscopy methods. With the use of liquid cell, illumination system, and lowered electron dose the successful light-induced in-situ observations on Staphylococcus aureus encapsulated with methylene blue were performed. Results showed that with specified imaging parameters it is possible to conduct reliable research on bacteria in electron microscope despite the unfavorable damaging effect of the highly energetic electron beam used for imaging. This approach differs from the common methods, as it provides direct observations of the processes occurring upon light illumination. The effects obtained with the proposed method are very promising and may serve to answer why different microorganisms respond to the therapy differently.

Variability of Amyloid Propensity in Imperfect Repeats of CsgA Protein of Salmonella enterica and Escherichia coli.

CsgA is an aggregating protein from bacterial biofilms, representing a class of functional amyloids. Its amyloid propensity is defined by five fragments (R1-R5) of the sequence, representing non-perfect repeats. Gate-keeper amino acid residues, specific to each fragment, define the fragment's propensity for self-aggregation and aggregating characteristics of the whole protein. We study the self-aggregation and secondary structures of the repeat fragments of Salmonella enterica and Escherichia coli and comparatively analyze their potential effects on these proteins in a bacterial biofilm. Using bioinformatics predictors, ATR-FTIR and FT-Raman spectroscopy techniques, circular dichroism, and transmission electron microscopy, we confirmed self-aggregation of R1, R3, R5 fragments, as previously reported for Escherichia coli, however, with different temporal characteristics for each species. We also observed aggregation propensities of R4 fragment of Salmonella enterica that is different than that of Escherichia coli. Our studies showed that amyloid structures of CsgA repeats are more easily formed and more durable in Salmonella enterica than those in Escherichia coli.