Fibrous Aggregates Research Articles

Comprehensive review of protein imaging with AIEgens conjugated probes: From concentration to conformation

Proteins are an essential component of living organisms, and the study of their structure and function is of great importance in biological and medical research. In recent years, the remarkable phenomenon of aggregation-induced emission (AIE) has been extensively utilized in protein detection. Significant progress has been made in the development of aggregation-induced emission luminogens (AIEgens), which have proven invaluable in protein imaging. This review highlights AIEgen-conjugated probes for imaging proteins in tumor cells through various mechanisms, including physical interactions, ligand binding, and enzymatic cleavage. These probes exploit the AIE effect to enhance the signal-to-noise ratio, providing important tools for protein research. Additionally, these probes can be used to study structural changes in intracellular protein phase separation processes, such as unfolded, misfolded, fibrous, and amorphous aggregates. The above research achievements presented lay the foundation for the widespread application of AIEgen-conjugated probes in the biomedical field and are expected to stimulate further development in related research.

Hydroxyl group dynamics in defernite: Raman spectroscopy studies

A detailed examination of the altered silicate-carbonate xenolith embedded within the ignimbrite of the Upper Chegem Caldera revealed a new occurrence of a rare carbonate mineral known as defernite, with chemical formula Ca6[(CO3)2-x(Si2O7)x/2](OH)7[Cl1-x(H2O)x], where x ≈ 0.4. Defernite crystallizes as colorless to white fibrous aggregates, reaching 100–150 μm diameters. Subsequently, Raman investigations of defernite from the Upper Chegem Caldera were conducted to perform a comprehensive structural analysis and compare it with minerals found in other locations. During this examination, band assignments focused on the carbonate ion vibration (CO32−) with a band at 1085 cm−1 and the hydroxyl group, characterized by a series of strong bands around 3590–3600 cm−1, particularly evident in oriented crystals along the (010) plane. Experimentation involving the alteration of incident laser light polarization highlighted a reduction in the intensity of carbonate and hydroxyl-related bands and the activation of a band around 3390 cm−1. This phenomenon is explained by the formation of hydrogen bonding between hydroxyl groups and chlorine or molecular water, potentially occupying chlorine positions. Lastly, a temperature-dependent experiment demonstrated the instability of the 3390 cm−1 band, which dissipated with increasing temperature. This insight explains the band’s origin around 3590 cm−1, ascribed to non-degenerate hydroxyl groups as a key marker within the defernite structure.

Osmotic stress induces formation of both liquid condensates and amyloids by a yeast prion domain

Liquid protein condensates produced by phase separation are involved in the spatiotemporal control of cellular functions, while solid fibrous aggregates (amyloids) are associated with diseases and/or manifest as infectious or heritable elements (prions). Relationships between these assemblies are poorly understood. The Saccharomyces cerevisiae release factor Sup35 can produce both fluid liquid-like condensates (e. g. at acidic pH) and amyloids (typically cross-seeded by other prions). We observed acidification-independent formation of Sup35-based liquid condensates in response to hyperosmotic shock in the absence of other prions, both at increased and physiological expression levels . The Sup35 prion domain, Sup35N, is both necessary and sufficient for condensate formation, while the middle domain, Sup35M antagonizes this process. Formation of liquid condensates in response to osmotic stress is conserved within yeast evolution. Notably, condensates of Sup35N/NM protein originated from the distantly related yeast Ogataea methanolica can directly convert to amyloids in osmotically stressed S. cerevisiae cells, providing a unique opportunity for real-time monitoring of condensate-to-fibril transition in vivo by fluorescence microscopy. Thus, cellular fate of stress-induced condensates depends on protein properties and/or intracellular environment.

Savelievaite, Mg2CrO2(BO3), the first natural borate with species-defining Cr3+ and the ludwigite–savelievaite isomorphous series

AbstractThe new ludwigite-group mineral savelievaite, ideally Mg2Cr3+O2(BO3), was found in the chromitite body at the Malaya Kharamatalou river valley, Voikar–Syninskiy ultrabasic complex, Polar Urals, Russia. Savelievaite and Cr-enriched ludwigite occur in clinochlore veinlets and are associated with earlier magnesiochromite, spinel, chromite, pargasite, diopside, forsterite, serpentine, magnetite and pentlandite. Savelievaite forms prismatic, acicular or fibrous crystals up to 0.05 × 0.4 mm, usually assembled in radiating or chaotic clusters up to 1 × 1.5 mm across. It is opaque, black to greenish-black. The lustre is vitreous for prismatic crystals and silky for fibrous aggregates. D(calc.) = 3.91 g cm–3. Under the microscope in reflected light, savelievaite is grey, non-pleochroic, with weak bireflectance and anisotropism. The chemical composition (wt.%, EMPA, Fe2+:Fe3+ ratio by stoichiometry) is: MgO 34.88, FeO 10.83, NiO 0.36, B2O3 16.80, Al2O3 2.97, V2O3 0.21, Cr2O3 21.97, Fe2O3 12.40, TiO2 0.43, total 100.85. The empirical formula calculated on the basis of 5 O apfu is (Mg1.72Fe2+0.30Ni0.01)Σ2.03(Cr3+0.57Fe3+0.31Al0.12Ti0.01V3+0.01)Σ1.02B0.96O5. Savelievaite is orthorhombic, space group Pbam, a = 9.2631(6), b = 12.2298(8), c = 3.0104(2) Å, V = 341.04(4) Å3 and Z = 4. The strongest reflections of the powder X-ray diffraction pattern [d,Å(I)(hkl)] are: 5.101(100)(120); 2.551(90)(240); 2.524(88)(201); 2.163(36)(250); and 2.033(55)(321). The crystal structure was solved from single-crystal X-ray diffraction data and refined to R1 = 0.0405. Savelievaite is isostructural with ludwigite, Cr3+ is concentrated at the M4 site. The mineral is named in honour of the Russian petrologist and geologist Dr. Galina Nikolaevna Savelieva (b. 1936). Ludwigite, ideally Mg2Fe3+O2(BO3), and savelievaite form a continuous isomorphous series in which Cr3+ content varies from 0 to 0.60 apfu. Occurrences of Cr-enriched (>1 wt.% Cr2O3) varieties of ludwigite are mainly related to ultrabasic complexes. The Cr-richest (>10 wt.% Cr2O3) ludwigite–savelievaite-series members are found in chromite ores at the Voikar–Syninskiy complex and Volchiegorskoe and Tatishchevskoe deposits, both in the South Urals.

Clinical and Pathological Features of FTDP-17 with MAPT p.K298_H299insQ Mutation.

MAPT is a causative gene in frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), a hereditary degenerative disease with various clinical manifestations, including progressive supranuclear palsy, corticobasal syndrome, Parkinson's disease, and frontotemporal dementia. To analyze genetically, biochemically, and pathologically multiple members of two families who exhibited various phenotypes of the disease. Genetic analysis included linkage analysis, homozygosity haplotyping, and exome sequencing. We conducted tau protein microtubule polymerization assay, heparin-induced tau aggregation, and western blotting with brain lysate from an autopsy case. We also evaluated abnormal tau aggregation by using anti-tau antibody and PM-PBB3. We identified a variant, c.896_897insACA, p.K298_H299insQ, in the MAPT gene of affected patients. Similar to previous reports, most patients presented with atypical parkinsonism. Biochemical analysis revealed that the mutant tau protein had a reduced ability to polymerize microtubules and formed abnormal fibrous aggregates. Pathological study revealed frontotemporal lobe atrophy, midbrain atrophy, depigmentation of the substantia nigra, and four-repeat tau-positive inclusions in the hippocampus, brainstem, and spinal cord neurons. The inclusion bodies also stained positively with PM-PBB3. This study confirmed that the insACA mutation caused FTDP-17. The affected patients showed symptoms resembling Parkinson's disease initially and symptoms of progressive supranuclear palsy later. Despite the initial clinical diagnosis of frontotemporal dementia in the autopsy case, the spread of lesions could explain the process of progressive supranuclear palsy. The study of more cases in the future will help clarify the common pathogenesis of MAPT mutations or specific pathogeneses of each mutation.

Disaggregation of amyloid-beta fibrils via natural metabolites using long timescale replica exchange molecular dynamics simulation studies.

Amyloid fibrils are self-assembled fibrous protein aggregates that are associated with several presently incurable diseases such as Alzheimer's. disease that is characterized by the accumulation of amyloid fibrils in the brain, which leads to the formation of plaques and the death of brain cells. Disaggregation of amyloid fibrils is considered a promising approach to cure Alzheimer's disease. The mechanism of amyloid fibril formation is complex and not fully understood, making it difficult to develop drugs that can target the process. Diacetonamine and cystathionine are potential lead compounds to induce disaggregation of amyloid fibrils. In the current research, we have used long timescale molecular simulation studies and replica exchange molecular dynamics (REMD) for 1000 ns (1 μs) to examine the mechanisms by which natural metabolites can disaggregate amyloid-beta fibrils. Molecular docking was carried out using Glide and with prior protein minimization and ligand preparation. We focused on a screening a database of natural metabolites, as potential candidates for disaggregating amyloid fibrils. We used Desmond with OPLS 3e as a force field. MM-GBSA calculations were performed. Blood-brain barrier permeability, SASA, and radius of gyration parameters were calculated.

Supergene mineral assemblage from sediments affected by contact and hydrothermal metamorphism, locality Dobrá – Staré Město near Frýdek-Místek (Czech Republic)

At locality Dobrá – Staré Město, series of outcrops occur on both banks of the Morávka River. The river cuts through the sediments on the tectonic contact between the Silesian and Subsilesian units of the Outer Western Carpathians. Clayey facies of the Hradiště Formation (Early Cretaceous, Valanginian to Aptian) contains numerous tectonic fragments reaching hundreds meters in size of strongly altered (carbonatized, smectitized) subaquatic volcanic rocks of the Teschenite Association. These volcanic rocks are dominated by fine-grained to aphanitic rocks, possibly of both effusive and intrusive origin. The host sediments affected by the contact metamorphism (contact adinole) also occur. Framboidal pyrite is abundant in majority of sediments of the studied area, giving rise to efflorescences of gypsum, baryte, celestine, and a mineral of alunite group (Figure 1). A more varied assemblage of supergene minerals was recognized on a small rock outcrop at GPS coordinates N 49° 39.992’ E 018° 23.810’ (Figure 2), where it developed relatively recently after the big floods in 2010 that washed away the previous mineralization. During the first years, only gypsum crusts were detectable, while in 2023, we recognized 4 macroscopic and 5 microscopic minerals forming thin botryoidal crusts. These crusts show faint zoning, with gooey, gel-like X-ray amorphous material in the humid upper part , middle zone with abundant fibroferrite and the lower part dominated by gypsum. Gypsum forms well-developed, but frequently corroded crystals with various habitus (Figure 4A, B). Fibroferrite is present as fibrous aggregates with individual crystals of up to ca. 100–150 µm long and <1 µm thick (Figure 4C). It forms directly from the gel-like substance (Figure 4D). Rhombohedral crystals and rossete aggregates of hydroniumjarosite vary from 2 µm to 5 µm in size (Figure 4E), but the most common are dust-like coatings on rock fissures with particle size <1 µm. Rusty coatings of schwertmannite (Figure 4F), in some places accompanied by gypsum, with Fe/S at. % ratios near 4–5 are common. In some cases, the Al/Fe at. % ratio exceed 1, which might indicate presence of yet undescribed Al-analogue of schwertmannite, ill-defined alumogel, or an amorphous precursor of aluminite. Presence of ferrihydrite is possible but not confirmed. The above-mentioned Fe-rich phases are frequently accompanied by diplobacilli-shaped bacteria (Figure 4F). Slavíkite, pickeringite-halotrichite, alunogen, copiapite group mineral, hexahydrite, and melanterite (Figure 5) were also recognized by EDS and PXRD. Slavíkite and copiapite group minerals seem to be alteration products of fibroferrite. Unit-cell parameters refined from the X-ray powder diffraction are presented for all recognized phases (Table 1). Surprisingly, secondary sulfates are present on the metasediments with abundant calcite (Figure 3), which should serve as a neutralization buffer. The reason is the restricted amount of calcite available on the rock fissure surfaces, and water infiltration partially blocked by newly-formed gypsum. The albite in contact adinole (Figure 3) is unaffected by the acid rock drainage, since Na ions are not present in significant quantities in supergene phases. Since there is still enough available unaltered pyrite, we expect a shift towards more stable mineral assemblage in future. The limited amount of secondary sulfates does not present any significant harm to the environment.

Molecular mechanisms involved in the destabilization of two types of R3-R4 tau fibrils associated with chronic traumatic encephalopathy by Fisetin.

Chronic traumatic encephalopathy is a neurodegenerative tauopathy pathologically characterized by fibrillary tau aggregates in the depth of sulci. Clearing fibrous tau aggregates is considered a promising strategy in the treatment of CTE. Fisetin (FS), a natural polyphenolic small molecule, was confirmed to disassociate the tau filaments in vitro. However, the molecular mechanisms of FS in destabilizing the CTE-related R3-R4 tau fibrils remain largely unknown. In this study, we compared the atomic-level structural differences of the two types of CTE-related R3-R4 tau fibrils and explored the influence and molecular mechanisms of FS on the two types of fibrils by conducting multiple molecular dynamics (MD) simulations. The results reveal that the type 1 fibril displays higher structural stability than the type 2 fibril, with a lower root-mean-square-fluctuation value and higher β-sheet structure probability. FS can destabilize both types of fibrils by decreasing the β-sheet structure content, interrupting the mainchain H-bond network, and increasing the solvent accessible surface area and β7-β8 angle of the fibrils. H-bonding, π-π stacking and cation-π are the common interactions driving FS molecules binding on the two types of fibrils, while the hydrophobic interaction occurs only in the type 2 fibril. Due to the relatively short simulation time, our study captures the early molecular mechanisms. However, it does provide beneficial information for the design of drugs to prevent or treat CTE.

MINERAL COMPOSITION AND FORMATION CONDITIONS OF BOTTOM SEDIMENTS OF THE SMALL LAKES IF THE UKOK PLATEAU (ALTAI)<a href="#FN1"><sup>1</sup></a>

Numerous modern lakes appeared in the intermountain basins and on the high-elevation plateaus of Altai within the immediate distribution of glaciation in the Neo-Pleistocene. There is little known about the geochemical processes of autigenic mineral formation in lake systems under conditions of nival sedimentogenesis. The study of cores of bottom sediments of 8 high-altitude lakes of Altai showed that the waters of the lakes are fresh bicarbonate with variations in the cationic composition of Ca–Na. The bottom sediments of lakes are characterized by different ratios of mineral detrital material, autigenic minerals (calcite, gypsum, pyrite, illite) and mortmass of plant residues. In the bottom sediments of lakes located within the same basin (Bertek or Tarkhatinskaya), the absolute concentrations of elements differ within one standard deviation, with the exception of significant variations in the contents of individual elements (Mo, U, Li, Be). The enrichment of bottom sediments of lakes (Argamdzhi, Teply Klyuch, Krasnoe) with these elements is associated with the presence of ore concentrations (Mo, U, Li, Be) in the local catchment areas of a single lake. The mineral associations of the sediments of the studied lakes differ from each other in the composition of autigenic minerals, and from rocks, soils of catchment areas in the composition of layered silicates. In the composition of fine-scaled, tangled fibrous aggregates of the bottom sediment illite, the amount of iron is 2-4 times greater than in the composition of lamellar aggregates of micas, chlorites from rocks, soils of catchment areas. Cryogenic processes have determined the crystallization of gypsum nodules in the lakes Argamdzhi and Small Tarkhatinskoye; and calcite aggregates in Zerlyukol-Nur and Large Tarkhatinskoye in significant volumes for lakes with fresh water composition.

Precise control of aggregation morphology: Effective strategy to tune the properties of ovotransferrin particles

Different aggregation morphologies of ovotransferrin (OVT) aggregates were successfully obtained through precise control, and the effects on structural, physical, liquid–liquid and gas–liquid interfacial characteristics as well as mechanisms were explored for the first time. It was observed that the surface hydrophobicity of OVT fibrils was higher than OVT spheres due to the acid-heat treatment. The exploration of liquid–liquid interface behaviors indicated that OVT fibrils possessed higher adsorption capacity at the interface, revealing the higher surface activity at the oil-water interface. During adsorption process, fibrils exhibited higher diffusion rate, while spheres were easier to penetrate and rearrange at the interface. The interfacial film composed of fibrils possessed more elastic solid-like behaviors owing to the higher surface activity of individual fibrous aggregates and rapid fibril-fibril interactions. The analysis of gas–liquid interface characteristics presented that OVT spheres possessed lower interfacial tension and higher interfacial viscoelasticity, and showed significantly higher FC and FS values in comparation to fibrils. These findings will facilitate the reader's understanding of the relationship between protein aggregate structure and properties, and lay a foundation for broadening the application of OVT and even other proteins.

Chemical-induced phase transition and global conformational reorganization of chromatin

Chemicals or drugs can accumulate within biomolecular condensates formed through phase separation in cells. Here, we use super-resolution imaging to search for chemicals that induce phase transition within chromatin at the microscale. This microscopic screening approach reveals that adriamycin (doxorubicin) — a widely used anticancer drug that is known to interact with chromatin — specifically induces visible local condensation and global conformational change of chromatin in cancer and primary cells. Hi-C and ATAC-seq experiments systematically and quantitatively demonstrate that adriamycin-induced chromatin condensation is accompanied by weakened chromatin interaction within topologically associated domains, compartment A/B switching, lower chromatin accessibility, and corresponding transcriptomic changes. Mechanistically, adriamycin complexes with histone H1 and induces phase transition of H1, forming fibrous aggregates in vitro. These results reveal a phase separation-driven mechanism for a chemotherapeutic drug.

Macrophage membrane-encapsulated nitrogen-doped carbon quantum dot nanosystem for targeted treatment of Alzheimer's disease: Regulating metal ion homeostasis and photothermal removal of β-amyloid

The abnormal aggregation of β-amyloid protein (Aβ) is a major contributor to Alzheimer's disease (AD). Cu2+ homeostasis imbalance can lead to the aggregation of Aβ, resulting in cytotoxic oligomers and fibrous aggregates, causing neuroinflammation and nerve cell damage, ultimately leading to AD. In this study, we synthesized nitrogen-doped carbon quantum dot (CQD), and designed a macrophage membrane (RAW-M) encapsulated CQD nanosystem for the first time. The abundant nitrogen-containing groups on the surface of CQD effectively capture excess Cu2+ and inhibit rapid Aβ aggregation. Additionally, the good photothermal properties of CQD dissolve the formed fiber precipitates under near-infrared light (NIR). In vitro and in vivo studies showed that the nanosystem significantly improved BBB permeability under laser irradiation, enhancing its ability to cross the BBB and overcome traditional anti-AD drug limitations. In vivo investigations conducted on APP/PS1 mice indicate that the nanosystem strongly reduced Aβ deposition, mitigated neuroinflammation, and ameliorates deficits in learning and memory. Overall, our nanocarrier approach adjusts metal ion homeostasis, inhibits abnormal Aβ aggregation, and uses excellent photothermal properties to depolymerize mature Aβ fibrils to protect cells from Aβ neurotoxicity, providing an effective strategy for Aβ-targeted treatment of AD.

Yeast Chaperone Hsp70-Ssb Modulates a Variety of Protein-Based Heritable Elements.

Prions are transmissible self-perpetuating protein isoforms associated with diseases and heritable traits. Yeast prions and non-transmissible protein aggregates (mnemons) are frequently based on cross-β ordered fibrous aggregates (amyloids). The formation and propagation of yeast prions are controlled by chaperone machinery. Ribosome-associated chaperone Hsp70-Ssb is known (and confirmed here) to modulate formation and propagation of the prion form of the Sup35 protein [PSI+]. Our new data show that both formation and mitotic transmission of the stress-inducible prion form of the Lsb2 protein ([LSB+]) are also significantly increased in the absence of Ssb. Notably, heat stress leads to a massive accumulation of [LSB+] cells in the absence of Ssb, implicating Ssb as a major downregulator of the [LSB+]-dependent memory of stress. Moreover, the aggregated form of Gγ subunit Ste18, [STE+], behaving as a non-heritable mnemon in the wild-type strain, is generated more efficiently and becomes heritable in the absence of Ssb. Lack of Ssb also facilitates mitotic transmission, while lack of the Ssb cochaperone Hsp40-Zuo1 facilitates both spontaneous formation and mitotic transmission of the Ure2 prion, [URE3]. These results demonstrate that Ssb is a general modulator of cytosolic amyloid aggregation, whose effect is not restricted only to [PSI+].

Amphipathic peptide–phospholipid nanofibers: Kinetics of fiber formation and molecular transfer between assemblies

Understanding the kinetics of nano-assembly formation is important to elucidate the biological processes involved and develop novel nanomaterials with biological functions. In the present study, we report the kinetic mechanisms of nanofiber formation from a mixture of phospholipids and the amphipathic peptide 18A[A11C], carrying cysteine substitution of the apolipoprotein A-I-derived peptide 18A at residue 11. 18A[A11C] with acetylated N-terminus and amidated C-terminus can associate with phosphatidylcholine to form fibrous aggregates at neutral pH and lipid-to-peptide molar ratio of ∼1, although the reaction pathways of self-assembly remain unclear. Here, the peptide was added to giant 1-palmitoyl-2-oleoyl phosphatidylcholine vesicles to monitor nanofiber formation under fluorescence microscopy. The peptide initially solubilized the lipid vesicles into particles smaller than the resolution of optical microscope, and fibrous aggregates appeared subsequently. Transmission electron microscopy and dynamic light scattering analyses revealed that the vesicle-solubilized particles were spherical or circular, measuring ∼10–20 nm in diameter. The rate of nanofiber formation of 18A with 1,2-dipalmitoyl phosphatidylcholine from the particles was proportional to the square of lipid–peptide concentration in the system, suggesting that the association of particles, accompanied by conformational changes, was the rate-limiting step. Moreover, molecules in the nanofibers could be transferred between aggregates faster than those in the lipid vesicles. These findings provide useful information for the development and control of nano-assembling structures using peptides and phospholipids.

A double point mutation of SOD1 targeting net charge promotes aggregation under destabilizing conditions: Correlation of charge distribution and ALS-provoking mutation

A progressive neurodegenerative illness such as amyotrophic lateral sclerosis (ALS) is characterized by the misfolding and aggregation of human CuZn superoxide dismutase (hSOD1) into amyloid aggregates. Thus, designing strategies for the choice of WT-SOD1 and double mutant (G12D/G138E) with an increased net negative charge can be a good idea to elucidate the pathological mechanism of SOD1 in ALS under some destabilizing conditions. Consequently, we show evidence that protein charge, together with other destabilizing conditions, plays an important role in ALS disease. To achieve this purpose, we use methods, such as spectroscopy and transmission electron microscopy (TEM) to monitor the formation of amyloid aggregation. The specific activity of WT-SOD1 was approximately 1.72 times higher than that of the double mutant. Under amyloidogenic circumstances, structural properties such as local, secondary, oligomeric, and fibrillar structures were explored. The double mutant's far-UV CD spectra displayed a broad minimum peak in the region 213 to 218 nm, suggesting the production of β-rich amyloid fibrils. FTIR spectra of the double mutant samples at different incubation times showed a low-frequency peak around 1630–1640 cm-1, attributed to a parallel β-sheet. Moreover, CR-binding assay and TEM analysis revealed and confirmed that mutation with an increased repulsive charge promotes the formation of fibrous aggregates. Consequently, ALS mutations with a higher repulsive charge are the apparent exceptions that validate the rule. This findings revealed that the double mutant increases protein aggregation through a novel mechanism, likely involving destabilization of structure and a change in the net negative charge.

High-internal-phase emulsions stabilized solely by chitosan hydrochloride: Fabrication and effect of pH on stabilization mechanism

High-internal-phase emulsions (HIPEs) have numerous applications, particularly in the food industry, and are therefore extensively researched. In particular, the discovery of green, simple, and efficient HIPE stabilizers and the elucidation of their structure-activity relationships and stabilization mechanisms are of great practical significance. Herein, chitosan hydrochloride (CHC) was used to stabilize variable-transparency HIPEs under different conditions. At a CHC content of 0.08 wt.%, one-step shear dispersion at pH 6.0–6.7 produced milky white HIPEs with small droplets (∼5 μm), high stability, strong gelation, and good plasticity, whereas highly transparent HIPEs with larger droplets (∼100 μm) were formed at pH 6.8–12.7. The stabilization mechanism depended on pH, corresponding to the formation of (i) three-dimensional (3D) reticular membranes (pH 6.0–6.7) or (ii) fibrous Pickering particles (pH 6.8–12.7). This dependence was ascribed to the effects of pH on CHC structure: CHC mainly existed as a nanofiber network with grids at pH 6.0–6.7, whereas fibrous aggregates were observed at higher pH. Thus, our study presents important insights into the pH-dependent mechanism of HIPE stabilization by CHC and facilitates the development of new emulsifiers for the formulation of variable-transparency HIPEs with diverse applications.

Amyloid Fibrils Enhance the Topical Bio-Adhesivity of Liquid Crystalline Mesophase-Based Drug Formulations.

Despite their distinctive secondary structure based on cross β-strands, amyloid fibrils (AF) are stable fibrous protein aggregates with features similar to collagen, one of the main components of the extracellular matrix, and thus constitute a potential scaffold for enhancing cell adhesion for topical applications. Here, the contribution of AF to skin bio-adhesivity aiming toward topical treatments is investigated. Liquid crystalline mesophase (LCM) based on phytantriol is formulated, with the aqueous phase containing either water or a solution of 4wt% amyloid fibrils. Then resveratrol is added as a model anti-inflammatory molecule. The developed LCM presents a double gyroid Ia3d mesophase. The incorporation of AF into the LCM increases its bio-adhesive properties. In vitro release and ex vivo permeation and retention confirm the controlled release property of the system, and that resveratrol is retained in epidermis and dermis, but is also permeated through the skin. All formulations are biocompatible with L929 cells. The in vivo assay confirms that systems with AF lead to a higher anti-inflammatory effect of resveratrol. These results confirm the hypothesis that the incorporation of AF in the LCM increases the bio-adhesiveness and efficiency of the system for topical treatment, and consequently, the therapeutical action of the encapsulated drug.

Retgersite from mine dump of the Lill shaft, Březové Hory ore district, Příbram (Czech Republic)

Rare sulphate of nickel, retgersite, was identified on the one specimen collected on the dump of the Lill shaft, Březové Hory ore district, Příbram (Czech Republic) in 1990s. The specimen consists of quartz gangue with dominant millerite and sphalerite. Retgersite forms light blue to blue-green curved fibrous microcrystalline aggregates up to 2 mm in size with vitreous lustre, growing on the surface and in small outer vugs of the supergene altered gangue. Retgersite was identified by PXRD and its refined unit-cell parameters (for the tetragonal space group P41212) are: a 6.7838(2) Å, c 18.2951(8) Å and V 841.94(4) Å3. Results of semiquantitative chemical analysis (EDS) as well as vibrational spectra (Raman, infrared) including their tentative assignments are also given. The studied retgersite was formed by (sub)recent weathering of primary nickel mineralisation (mostly millerite) in the mine dump conditions.

Origin of stronger binding of ionic pair (IP) inhibitor to Aβ42 than the equimolar neutral counterparts: synergy mechanism of IP in disrupting Aβ42 protofibril and inhibiting Aβ42 aggregation under two pH conditions.

Fibrous aggregates of beta-amyloid (Aβ) is a hallmark of Alzheimer's disease (AD). Several major strategies of drugs or inhibitors, including neutral molecules, positive or negative ions, and dual-inhibitor, are used to inhibit the misfolding or aggregation of Aβ42, among which a kind of dual-inhibitor composed of a pair of positive and negative ions is emerging as the most powerful candidate. This knowledge lacks the origin of the strong inhibitory effect and synergy mechanisms blocking the development and application of such inhibitors. To this end, we employed 1 : 1 ionic pairs (IP) of oppositely charged benzothiazole molecules (+)BAM1-EG6 (Pos) and (-)BAM1-EG6 (Neg) as well as equimolar neutral BAM1-EG6 (Neu) counterpart at two pH conditions (5.5 and 7.0) to bind Aβ42 targets, Aβ42 monomer (AβM), soluble pentamer (AβP), and pentameric protofibril (AβF) models, respectively, corresponding to the products of three toxic Aβ42 development pathways, lag, exponential and fibrillation phases. Simulated results illustrated the details of the inhibitory mechanisms of IP and Neu for the AβY (Y = M, P, or F) in the three different phases, characterizing the roles of Pos and Neg of IP as well as their charged, hydrophobic groups and linker playing in the synergistic interaction, and elucidated a previously unknown molecular mechanism governing the IP-Aβ42 interaction. Most importantly, we first revealed the origin of the stronger binding of IP inhibitors to Aβ42 than that of the equimolar neutral counterparts, observing a perplexing phenomenon that the physiological condition (pH = 7.0) than the acidic one (pH = 5.5) is more favorable to the enhancement of IP binding, and finally disclosed that solvation is responsible to the enhancement because at pH 7.0, AβP and AβF act as anionic membranes, where solvation plays a critical role in the chemoelectromechanics. The result not only provides a new dimension in dual-inhibitor/drug design and development but also a new perspective for uncovering charged protein disaggregation under IP-like inhibitors.

A liquid-to-solid phase transition of Cu/Zn superoxide dismutase 1 initiated by oxidation and disease mutation

Cu/Zn superoxide dismutase 1 (SOD1) has a high propensity to misfold and form abnormal aggregates when it is subjected to oxidative stress or carries mutations associated with amyotrophic lateral sclerosis. However, the transition from functional soluble SOD1 protein to aggregated SOD1 protein is not completely clear. Here, we propose that liquid-liquid phase separation (LLPS) represents a biophysical process that converts soluble SOD1 into aggregated SOD1. We determined that SOD1 undergoes LLPS invitro and cells under oxidative stress. Abnormal oxidation of SOD1 induces maturation of droplets formed by LLPS, eventually leading to protein aggregation and fibrosis, and involves residues Cys111 and Trp32. Additionally, we found that pathological mutations in SOD1 associated with ALS alter the morphology and material state of the droplets and promote the transformation of SOD1 to solid-like oligomers which are toxic to nerve cells. Furthermore, the fibrous aggregates formed by both pathways have a concentration-dependent toxicity effect on nerve cells. Thus, these combined results strongly indicate that LLPS may play a major role in pathological SOD1 aggregation, contributing to pathogenesis in ALS.