Formation of self-assembled fibril aggregates of trypsin-controllably hydrolyzed soy protein and its regulation on stability of high internal phase Pickering emulsions.

The mechanisms of trypsin hydrolysis time on the structure of soy protein hydrolysate fibril aggregates (SPHFAs) and the stability of SPHFAs-high internal phase Pickering emulsions (HIPPEs) were investigated. SPHFAs were prepared using soy protein hydrolysate (SPH) with different trypsin hydrolysis time (0 min-120 min) to stabilize SPHFAs-HIPPEs. The results showed that moderate trypsin hydrolysis (30 min, hydrolysis degree of 2.31 %) induced SPH unfolding and increased the surface hydrophobicity of SPH, thereby promoting the formation of flexible SPHFAs with maximal thioflavin T intensity and ζ-potential. Moreover, moderate trypsin hydrolysis improved the viscoelasticity of SPHFAs-HIPPEs, and SPHFAs-HIPPEs remained stable after storage at 25 °C for 80 d and heating at 100 °C for 1 h. Excessive trypsin hydrolysis (> 30 min) decreased the stability of SPHFAs-HIPPEs. In conclusion, moderate trypsin hydrolysis promoted the formation of flexible SPHFAs with high surface charge by inducing SPH unfolding, thereby promoting the stability of SPHFAs-HIPPEs.

Fabrication of bamboo nanocellulose fibril-based food packaging with dual-antimicrobial property.

The development of cellulose-based packaging films with excellent antimicrobial properties and biocompatibility has garnered significant attention. In this work, nanocellulose fibrils (NCFs) derived from from bamboo parenchyma cells were utilized to fabricate nanocomposite film with antimicrobial properties. This system exhibited distinct release behaviors for two antimicrobial agents, with the slow release of Ag nanoparticle (AgNP) in the initial stage contributed to delaying food spoilage, while the subsequent pH change in the microenvironment facilitated the release of essential oil of sour orange blossoms (SEO) for secondary antimicrobial activity. Additionally, the composite film demonstrated improved thermal stability and UV blocking capacity. Moreover, AgNP has been proven to enhance the mechanical properties, with the tensile strength of the novel composite film increasing by 34.85 % compared to control group. The water vapor permeability and oxygen permeability of the novel composite film were reduced, which could potentially reduce weight loss and slow down the rate of after-ripening. Following the acidification treatment, the films containing EO@MPN (essential oil encapsulated with metal-polyphenol network) component performed different antimicrobial patterns, indicating their pH-responsive antimicrobial capabilities, and they are effective against both Gram-positive and Gram-negative bacteria. After a 24-h exposure to a food simulant, the release amount of Ag was measured at 67.6 µg/dm2, within the acceptable limit, and the release profile of Ag was characterized. Cytotoxicity and Live/Dead staining tests confirmed that the novel composite film film had no significant toxicity, thus making it safe for application in food preservation. Furthermore, in a 15-day preservation experiment with mangoes, the novel composite film demonstrated the best performance, underscoring its potential as a sustainable antimicrobial packaging material.

Amyloid nomenclature 2024: update, novel proteins, and recommendations by the International Society of Amyloidosis (ISA) Nomenclature Committee.

The ISA Nomenclature Committee met at the XIX International Symposium of Amyloidosis in Rochester, MN, 27 May 2024. The in-person event was followed by many electronic discussions, resulting in the current updated recommendations. The general nomenclature principles are unchanged. The total number of human amyloid fibril proteins is now 42 of which 19 are associated with systemic deposition, while 4 occur with either localised or systemic deposits. Most systemic amyloidoses are caused by the presence of protein variants which promote misfolding. However, in the cases of AA and ATTR the deposits most commonly consist of wild-type proteins and/or their fragments. One peptide drug, previously reported to create local iatrogenic amyloid deposits at its injection site, has been shown to induce rare instances of systemic deposition. The number of described animal amyloid fibril proteins is now 16, 2 of which are unknown in humans. Recognition of the importance of intracellular protein aggregates, which may have amyloid or amyloid-like properties, in many neurodegenerative diseases is rapidly increasing and their significance is discussed.

TDP-43 Amyloid Fibril Formation via Phase Separation-Related and -Unrelated Pathways.

Intrinsically disordered regions (IDRs) in proteins can undergo liquid-liquid phase separation (LLPS) for functional assembly, but this increases the chance of forming disease-associated amyloid fibrils. Not all amyloid fibrils form through LLPS however, and the importance of LLPS relative to other pathways in fibril formation remains unclear. We investigated this question in TDP-43, a motor neuron disease and dementia-causing protein that undergoes LLPS, using thioflavin T (ThT) fluorescence, NMR, transmission electron microscopy (TEM), and wide-angle X-ray scattering (WAXS) experiments. Using a fluorescence probe modified from ThT strategically designed for targeting protein assembly rather than ß-sheets and supported by TEM images, we propose that the biphasic ThT signals observed under LLPS-favoring conditions are due to the presence of amorphous aggregates. These aggregates represent an intermediate state that diverges from the direct pathway to ß-sheet-dominant fibrils. Under non-LLPS conditions in contrast (at low pH or at physiological conditions in a construct with key LLPS residues removed), the protein forms a hydrogel. Real-time WAXS data, ThT signals, and TEM images collectively demonstrate that the gelation process circumvents LLPS and yet still results in the formation of fibril-like structural networks. We suggest that the IDR of TDP-43 forms disease-causing amyloid fibrils regardless of the formation pathway. Our findings shed light on why both LLPS-promoting and LLPS-inhibiting mutants are found in TDP-43-related diseases.

Relationship between depth-wise refractive index and biomechanical properties of human articular cartilage.

Significance: Optical properties of biological tissues, such as refractive index (RI), are fundamental properties, intrinsically linked to the tissue's composition and structure. We hypothesize that, as the RI and the functional properties of articular cartilage (AC) are dependent on the tissue's structure and composition, the RI of AC is related to its biomechanical properties. Aim: This study aims to investigate the relationship between RI of human AC and its biomechanical properties. Approach: Human cartilage samples ( n = 22 ) were extracted from the right knee joint of three cadaver donors (one female, aged 47 years, and two males, aged 64 and 68 years) obtained from a commercial biobank (Science Care, Phoenix, Arizona, United States). The samples were initially subjected to mechanical indentation testing to determine elastic [equilibrium modulus (EM) and instantaneous modulus (IM)] and dynamic [dynamic modulus (DM)] viscoelastic properties. An Abbemat 3200 automatic one-wavelength refractometer operating at 600 nm was used to measure the RI of the extracted sections. Similarly, Spearman's and Pearson's correlation coefficients were employed for non-normal and normal datasets, respectively, to determine the correlation between the depth-wise RI and biomechanical properties of the cartilage samples as a function of the collagen fibril orientation. Results: A positive correlation with statistically significant relations ( p - values < 0.05 ) was observed between the RI and the biomechanical properties (EM, IM, and DM) along the tissue depth for each zone, e.g., superficial, middle, and deep zones. Likewise, a lower positive correlation with statistically significant relations ( p - values < 0.05 ) was also observed for collagen fibril orientation of all zones with the biomechanical properties. Conclusions: The results indicate that, although the RI exhibits different levels of correlation with different biomechanical properties, the relationship varies as a function of the tissue depth. This knowledge paves the way for optically monitoring changes in AC biomechanical properties nondestructively via changes in the RI. Thus, the RI could be a potential biomarker for assessing the mechanical competency of AC, particularly in degenerative diseases, such as osteoarthritis.

Properties of Skin Collagen from Southern Catfish (Silurus meridionalis) Fed with Raw and Cooked Food.

The southern catfish (Silurus meridionalis) is an economically important carnivorous freshwater fish in China. In this study, we compared the properties of skin collagen from southern catfish fed with raw food (RF) and cooked food (CF). The skin collagen yield in the RF group (8.66 ± 0.11%) was significantly higher than that of the CF group (8.00 ± 0.27%). SDS-PAGE, circular dichroism spectroscopy, and FTIR analyses revealed that the collagen extracted from southern catfish skin in both groups was type I collagen, with a unique triple helix structure and high purity. The thermal denaturation temperature of collagen in the RF group (35.20 ± 0.11 °C) was significantly higher than that of the CF group (34.51 ± 0.25 °C). The DPPH free radical scavenging rates were 68.30 ± 2.41% in the RF collagen and 61.78 ± 3.91% in the CF collagen, which was higher than that found in most fish collagen. Both the RF and CF groups had high ability to form fibrils in vitro. Under the same conditions, the CF group exhibited faster fibril formation and a thicker fibril diameter (p < 0.05). In addition, the RF group exhibited significantly higher expression of col1a1 compared to the CF group. These results indicated that feeding southern catfish raw food contributed to collagen production, and the collagen from these fish may have potential in biomaterial applications.

Morphological and Biophysical Study of S100A9 Protein Fibrils by Atomic Force Microscopy Imaging and Nanomechanical Analysis.

Atomic force microscopy (AFM) imaging enables the visualization of protein molecules with high resolution, providing insights into their shape, size, and surface topography. Here, we use AFM to study the aggregation process of protein S100A9 in physiological conditions, in the presence of calcium at a molar ratio 4Ca2+:S100A9. We find that S100A9 readily assembles into a worm-like fibril, with a period dimension along the fibril axis of 11.5 nm. The fibril's chain length extends up to 136 periods after an incubation time of 144 h. At room temperature, the fibril's bending stiffness was found to be 2.95×10-28 Nm2, indicating that the fibrils are relatively flexible. Additionally, the values obtained for the Young's modulus (Ex=6.96×105 Pa and Ey=3.37×105 Pa) are four orders of magnitude lower than those typically reported for canonical amyloid fibrils. Our findings suggest that, under the investigated conditions, a distinct aggregation mechanism may be in place in the presence of calcium. Therefore, the findings reported here could have implications for the field of biomedicine, particularly with regard to Alzheimer's disease.

Biophysical Studies of Amyloid-Binding Fluorophores to Tau AD Core Fibrils Formed without Cofactors.

Tau is an intrinsically disordered protein involved in several neurodegenerative diseases where a common hallmark is the appearance of tau aggregates in the brain. One common approach to elucidate the mechanisms behind the aggregation of tau has been to recapitulate in vitro the self-assembly process in a fast and reproducible manner. While the seeding of tau aggregation is prompted by negatively charged cofactors, the obtained fibrils are morphologically distinct from those found in vivo. The Tau AD core fragment (TADC, tau 306-378) has emerged as a new model and potential solution for the cofactor-free in vitro aggregation of tau. Here, we use TADC to further study this process combining multiple amyloid-detecting fluorophores and fibril bioimaging. We confirmed by transmission electron microscopy that this fragment forms fibrils after quiescent incubation at 37 °C. We then employed a panel of eight amyloid-binding fluorophores to query the formed species by acquiring their emission spectra. The results obtained showed that nearly all dyes detect TADC self-assembled species. However, the successful monitoring of TADC aggregation kinetics was limited to three fluorophores (X-34, Bis-ANS, and pFTAA) which yielded sigmoidal curves but different aggregation half-times, hinting to different species being detected. Altogether, this study highlights the potential of using multiple extrinsic fluorescent probes, alone or in combination, as tools to further clarify mechanisms behind the aggregation of amyloidogenic proteins.

Understanding Osaka mutation polymorphic Aß fibril response to static and oscillating electric fields: insights from computational modeling.

Alzheimer's disease (AD) is a prevalent neurodegenerative disorder, impacting millions of individuals worldwide. Among its defining characteristics is the accumulation of senile plaques within the brain's gray matter, formed through the self-assembly of misfolded proteins contributing to the progressive symptoms of AD. This study investigates a polymorphic Aß fibril under static and oscillating electric fields using molecular dynamics simulation. Specifically, we utilized a polymorphic fibrillar complex composed of two intertwined pentamer-strands of the Aß1-40 peptide with the Osaka mutation (E22Δ), known for its toxicity and stable structure. Our findings demonstrate that a 0.3 and 0.4 V/nm electric field combined with a 0.20 GHz frequency effectively disrupts the polymorphic conformation of Aß fibrils. Furthermore, we elucidate the molecular mechanisms underlying this disruption, providing insights into the potential therapeutic use of oscillating electric fields for AD. This research offers valuable insights into novel therapeutic approaches for combating AD pathology.

Study of Insulin Aggregation and Fibril Structure under Different Environmental Conditions.

Protein amyloid aggregation is linked with widespread and fatal neurodegenerative disorders as well as several amyloidoses. Insulin, a small polypeptide hormone, is associated with injection-site amyloidosis and is a popular model protein for in vitro studies of amyloid aggregation processes as well as in the search for potential anti-amyloid compounds. Despite hundreds of studies conducted with this specific protein, the procedures used have employed a vast array of different means of achieving fibril formation. These conditions include the use of different solution components, pH values, ionic strengths, and other additives. In turn, this variety of conditions results in the generation of fibrils with different structures, morphologies and stabilities, which severely limits the possibility of cross-study comparisons as well as result interpretations. In this work, we examine the condition-structure relationship of insulin amyloid aggregation under a range of commonly used pH and ionic strength conditions as well as solution components. We demonstrate the correlation between the reaction solution properties and the resulting aggregation kinetic parameters, aggregate secondary structures, morphologies, stabilities and dye-binding modes.

Amyloid Fibril Formation on Neuronal Cells in the Coexistence of Aß40 and Aß42.

The abnormal aggregation and subsequent deposition of amyloid ß-protein (Aß) in the brain are considered central to the pathogenesis of Alzheimer's disease. The two major species of Aß are Aß40 and Aß42, present at an approximate ratio of 9: 1. Accumulating evidence suggests that neuronal membranes are an important platform of amyloidogenesis by Aß. However, information on the aggregational behaviors of coexistent Aß40 and Aß42 on membranes is lacking. In this study, the aggregation and resultant cytotoxicity of coexistent Aß40 and Aß42 at a physiologically relevant ratio were investigated by fluorescence techniques. We found that the degree of coexistence of both Aßs in aggregates increased as the assembly proceeded, and reached a maximum in fibrils. However, the cytotoxicity of the mixed fibrils was weaker than that of Aß42 fibrils, indicating that Aß40 attenuates the toxicity of Aß42 by forming mixed fibrils. In contrast, the degree of coexistence was significantly lower in aqueous phase aggregation, highlighting different aggregation mechanisms between in membranes and in the aqueous phase.

Cryo-EM and solid state NMR together provide a more comprehensive structural investigation of protein fibrils.

The tropomyosin 1 isoform I/C C-terminal domain (Tm1-LC) fibril structure is studied jointly with cryogenic electron microscopy (cryo-EM) and solid state nuclear magnetic resonance (NMR). This study demonstrates the complementary nature of these two structural biology techniques. Chemical shift assignments from solid state NMR are used to determine the secondary structure at the level of individual amino acids, which is faithfully seen in cryo-EM reconstructions. Additionally, solid state NMR demonstrates that the region not observed in the reconstructed cryo-EM density is primarily in a highly mobile random coil conformation rather than adopting multiple rigid conformations. Overall, this study illustrates the benefit of investigations combining cryo-EM and solid state NMR to investigate protein fibril structure.

Advancing sustainable packaging through self-assembly induced amyloid fibrillization of soy and pea protein nanofilms.

This study explored protein fibrillization and characterization, demonstrating significant enhancements in the structural, mechanical, and functional properties of soy and pea protein fibrils for biodegradable food packaging. The fibrillizationprocess increased ß-sheet alignment by 1.3-fold for soy protein fibrils (SPF) and 1.2-fold for pea protein fibrils (PPF). ThT fluorescence assays revealed higher ß-sheet alignment in SPF compared to PPF. Structural analysis showed flexible, worm-like fibrils in SPF and PPF. Mechanical tests indicated significant improvements: tensile strength increased to 4.88 MPa for SPF and 3.83 MPa for PPF films, with elongation at break reaching 221 % for SPF and 101.62 % for PPF films. Amyloid fibrillation reduced water solubility and water vapor permeability while increasing the swelling degree of protein films. Optical analysis revealed decreased lightness, intensified green and yellow hues, and increased transparency. These findings highlight the potential of amyloid fibrillation to enhance protein films for sustainable packaging applications.

Fiber complex-stabilized high-internal-phase emulsion for allicin encapsulation: microstructure, stability, and thermal-responsive properties.

BACKGROUND: Stimuli-responsive emulsions have garnered significant attention for their ability to enhance sensory qualities and control the release of encapsulated nutrient in emulsion-based products. However, the characteristics of synthetic materials of fabricating stimuli-responsive emulsions have been a crucial limitation in the food industry. Regulating the behavior of molecules at the interface could potentially achieve the desired stimuli-responsive behavior, but currently there is limited information available. RESULTS: High-internal-phase emulsions (HIPEs) were fabricated for the encapsulation of allicin, stabilized by a complex of 20 g kg-1 whey protein amyloid fibrils (WPF) and 20 g kg-1 glycyrrhizin fibers (GA). The intermolecular interactions between WPF and GA in the fiber complexes were predominantly governed by hydrophobic and electrostatic forces. These complexes adsorbed and stacked around the oil droplets, forming a protective interfacial film that enhanced droplet stability. An increased proportion of WPF (WPF = 3:1 or 4:1) surrounding the oil droplets enhanced the accelerated storage stability of HIPEs, with instability indexes approaching 0.2. Additionally, HIPEs displayed a temperature-dependent modulus, with the emulsion stabilized by a WPF ratio of 3:1 showing the highest modulus at 85 °C. The encapsulation efficiency of allicin in HIPEs ranged from 88.69 ± 6.62% to 101 ± 1.37% at 25 °C, and from 31.95 ± 1.92% to 78.69 ± 4.63% after incubation at 85 °C for 8 h. The release profile of allicin from the HIPEs exhibited thermal responsiveness, depending on the interfacial content of GA. CONCLUSION: These findings indicated that the thermal-responsive properties of HIPEs can be strategically engineered by manipulating their interfacial characteristics. © 2024 Society of Chemical Industry.

Kilogram-scale production of strong and smart cellulosic fibers featuring unidirectional fibril alignment.

Multifunctional fibers with high mechanical strength enable advanced applications of smart textiles, robotics, and biomedicine. Herein, we reported a one-step degumming method to fabricate strong, stiff, and humidity-responsive smart cellulosic fibers from abundant natural grass. The facile process involves partially removing lignin and hemicellulose functioning as glue in grass, which leads to the separation of vessels, parenchymal cells, and cellulosic fibers, where cellulosic fibers are manufactured at kilogram scale. The resulting fibers show dense and unidirectional fibril structure at both micro- and nano-scales, which demonstrate high tensile strength of ∼0.9 GPa and Young's modulus of 72 GPa, being 13- and 14-times higher than original grass. Inspired by stretchable plant tendrils, we developed a humidity-responsive actuator by engineering cellulosic fibers into the spring-like structures, presenting superior response rate and lifting capability. These strong and smart cellulosic fibers can be manufactured at large scale with low cost, representing promising a fiber material derived from renewable and sustainable biomass.

Understanding alpha-synuclein aggregation propensity in animals and humans.

Alpha-synuclein (α-syn) aggregation plays a critical role in the pathogenicity of Parkinson's Disease (PD). This study aims to evaluate the aggregation propensity of α-syn fragment peptides designed using the variability found in humans and animals. Thioflavin T (ThT) and transmission electron microscopy (TEM) were used to validate the formation of fibrils to identify important amino acid residues. Human α-syn fragments 51-75, 37-61, 62-86, 76-100, and 116-140 demonstrate a significantly higher tendency to aggregate compared to fragments 1-25, 26-50, and 91-115. All species analyzed of the α-syn 37-61 and 62-86 regions were shown to form fibrils on both ThT and TEM. The α-syn 37-61 and 62-86 fragment regions exhibited a high susceptibility to aggregation, with fibril formation observed in all species. The A53T mutation in several α-syn 37-61 fragments may enhance their propensity for aggregation, suggesting a correlation between this mutation and the capacity for fibril formation. Furthermore, the presence of the non-amyloid-ß component (NAC) region, specifically in α-syn 62-86, was consistently observed in several fragments that displayed fibril formation, indicating a potential correlation between the NAC region and the process of fibril formation in α-syn. Finally, the combination of a high quantity of valine and a low quantity of acidic amino acids in these fragments may serve as indicators of α-syn fibril formation.

All-aqueous synthesis of alginate complexed with fibrillated protein microcapsules for membrane-bounded culture of tumor spheroids.

Water-in-water (W/W) emulsions provide bio-compatible all-aqueous compartments for artificial patterning and assembly of living cells. Successful entrapment of cells within a W/W emulsion via the formation of semipermeable capsules is a prerequisite for regulating on the size, shape, and architecture of cell aggregates. However, the high permeability and instability of the W/W interface, restricting the assembly of stable capsules, pose a fundamental challenge for cell entrapment. The current study addresses this problem by synthesizing multi-armed protein fibrils and controlling their assembly at the W/W interface. The multi-armed protein fibrils, also known as 'fibril clusters', were prepared by cross-linking lysozyme fibrils with multi-arm polyethylene glycol (PEG) via click chemistry. Compared to linear-structured fibrils, fibril clusters are strongly adsorbed at the W/W interface, forming an interconnected meshwork that better stabilizes the W/W emulsion. Moreover, when fibril clusters are complexed with alginate, the hybrid microcapsules demonstrate excellent mechanical robustness, semi-permeability, cytocompatibility and biodegradability. These advantages enable the encapsulation, entrapment and long-term culture of tumor spheroids, with great promise for applications for anti-cancer drug screening, tumor disease modeling, and tissue repair engineering.

Structural Reorganization Mechanism of the Aß42 Fibril Mediated by N-Substituted Oligopyrrolamide ADH-353.

The inhibition of amyloid-ß (Aß) fibrillation and clearance of Aß aggregates have emerged as a potential pharmacological strategy to alleviate Aß aggregate-induced neurotoxicity in Alzheimer's disease (AD). Maity et al. shortlisted ADH-353 from a small library of positively charged N-substituted oligopyrrolamides for its notable ability to inhibit Aß fibrillation, disintegrate intracellular cytotoxic Aß oligomers, and alleviate Aß-induced cytotoxicity in the SH-SY5Y and N2a cells. However, the molecular mechanism through which ADH-353 interacts with the Aß42 fibrils, leading to their disruption and subsequent clearance, remains unclear. Thus, a detailed molecular mechanism underlying the disruption of neurotoxic Aß42 fibrils (PDB ID 2NAO) by ADH-353 has been illuminated in this work using molecular dynamics simulations. Interestingly, conformational snapshots during simulation depicted the shortening and disappearance of ß-strands and the emergence of a helix conformation, indicating a loss of the well-organized ß-sheet-rich structure of the disease-relevant Aß42 fibril on the incorporation of ADH-353. ADH-353 binds strongly to the Aß42 fibril (ΔGbinding= -142.91 ± 1.61 kcal/mol) with a notable contribution from the electrostatic interactions between positively charged N-propylamine side chains of ADH-353 with the glutamic (Glu3, Glu11, and Glu22) and aspartic (Asp7 and Asp23) acid residues of the Aß42 fibril. This aligns well with heteronuclear single quantum coherence NMR studies, which depict that the binding of ADH-353 with the Aß peptide is driven by electrostatic and hydrophobic contacts. Furthermore, a noteworthy decrease in the binding affinity of Aß42 fibril chains on the incorporation of ADH-353 indicates the weakening of interchain interactions leading to the disruption of the double-horseshoe conformation of the Aß42 fibril. The illumination of key interactions responsible for the destabilization of the Aß42 fibril by ADH-353 in this work will greatly aid in designing new chemical scaffolds with enhanced efficacy for the clearance of Aß aggregates in AD.

Islet amyloid polypeptide fibril catalyzes amyloid-ß aggregation by promoting fibril nucleation rather than direct axial growth.

Aberrant aggregation of amyloid-ß (Aß) and islet amyloid polypeptide (IAPP) into amyloid fibrils underlies the pathogenesis of Alzheimer's disease (AD) and type 2 diabetes (T2D), respectively. T2D significantly increases AD risk, with evidence suggesting that IAPP and Aß co-aggregation and cross-seeding might contribute to the cross-talk between two diseases. Experimentally, preformed IAPP fibril seeds can accelerate Aß aggregation, though the cross-seeding mechanism remains elusive. Here, we computationally demonstrated that Aß monomer preferred to bind to the elongation ends of preformed IAPP fibrils. However, due to sequence mismatch, the Aß monomer could not directly grow onto IAPP fibrils by forming multiple stable ß-sheets with the exposed IAPP peptides. Conversely, in our control simulations of self-seeding, the Aß monomer could axially grow on the Aß fibril, forming parallel in-register ß-sheets. Additionally, we showed that the IAPP fibril could catalyze Aß fibril nucleation by promoting the formation of parallel in-register ß-sheets in the C-terminus between bound Aß peptides. This study enhances our understanding of the molecular interplay between Aß and IAPP, shedding light on the cross-seeding mechanisms potentially linking T2D and AD. Our findings also underscore the importance of clearing IAPP deposits in T2D patients to mitigate AD risk.

A Cationic Zn-Phthalocyanine Turns Alzheimer's Amyloid ß Aggregates into Non-Toxic Oligomers and Inhibits Neurotoxicity in Culture.

Amyloid ß peptide (Aß) aggregation and deposition are considered the main causes of Alzheimer's disease. In a previous study, we demonstrated that anionic Zn-phthalocyanine (ZnPc) can interact with the Aß peptide and inhibit the fibril-formation process. However, due to the inability of anionic ZnPc to cross the intact blood-brain barrier, we decided to explore the interaction of cationic methylated Zn-phthalocyanine (cZnPc) with the peptide. Using a ThT fluorescence assay, we observed that cZnPc dose-dependently and time-dependently inhibited Aß1-42 fibril levels under in vitro fibril-formation conditions. Electron microscopy revealed that it caused Aß1-42 peptides to form small aggregates. Western blotting and dot immunoblot oligomer experiments demonstrated that cZnPc increased rather than decreased the levels of oligomers from the very early stages of incubation. A binding assay confirmed that cZnPc could bind with the peptide. Docking simulations indicated that the oligomer species of Aß1-42 had a higher ability to interact with cZnPc. ANS fluorescence assay results indicated that cZnPc did not affect the hydrophobicity of the peptide. However, cZnPc significantly increased intrinsic tyrosine fluorescence of the peptide after 8 h of incubation in fibril-formation conditions. Importantly, cell culture experiments demonstrated that cZnPc did not exhibit any toxicity up to a concentration of 10 µM. Instead, it protected a neuronal cell line from Aß1-42-induced toxicity. Thus, our results suggest that cZnPc can affect the aggregation process of Aß1-42, rendering it non-toxic, which could be crucial for the therapy of Alzheimer's disease.