Maintaining the structure and functionality of proteins is crucial in applications ranging from food preservation to pharmaceutical formulation. Ethanol, while commonly used as a solvent and preservative, can induce structural changes in proteins depending on its concentration and the specific structure of the protein itself. This study investigates the structural effects of ethanol on three types of model proteins, namely bovine serum albumin (BSA), β-Lactoglobulin (β-Lg), and β-Casein (β-Cn), by using UV-Vis spectroscopy and fluorescence spectroscopy. The conformational responses of proteins in water-EtOH solutions of various ethanol concentrations (0-10 %, v/v) were analyzed through absorbance and emission spectral changes. At increasing ethanol concentration, UV absorption data showed distinct protein-dependent spectral changes. β-Lg and β-Cn exhibited strong hypochromism (an absorbance decrease of ∼25 %) and red-shifting at 222 nm and 220 nm, respectively, indicating partial unfolding and solvent exposure of aromatic residues. BSA demonstrated subtle changes, and consistent quenching in fluorescence with a continuous blue-shifting to 330 nm, suggesting a moderate overall stability and local rearrangements in its structure. β-Cn exhibited red-shifted fluorescence and quenching, reflecting its flexible, disordered structure and heterogeneous response to solvent conditions. Statistical analysis revealed that while fluorescence spectroscopy was highly sensitive to the intrinsic differences between proteins (p < 0.001), the ethanol-induced conformational changes were too subtle to be detected as a statistically significant treatment effect. The consistency of these trends indicates a rational underlying mechanism of interaction. This reflects the subtle nature of the effect at the tested concentrations rather than the absence of an effect. Moreover, these results unveil the protein-specific effects of ethanol and strongly emphasize the importance of solvent composition in maintaining protein integrity. Ethanol concentrations up to 5 % may offer protein stability whereas high ethanol levels (≥ 5-10 %) promote structural perturbations. These results will be useful for both basic scientific research, such as biophysical studies and the advancement of optical techniques, and various industrial uses.

RufO is a Cytochrome P450 enzyme involved in synthesising Rufomycin, a circular peptide with antibacterial activity. Herein, we present structural and biophysical analyses to resolve the ambiguity of RufO's substrate specificity. The structure of unliganded RufO, alongside a series of computational and biophysical studies investigating its substrate specificity in the presence of ferredoxin, which is known to serve as an effector of the redox activities of several P450 enzymes. Contrary to reports on RufO's catalytic activity, monomeric L-tyrosine was not recognized by RufO in our isothermal titration calorimetry (ITC) experiments. Instead, RufO recognizes a range of putative substrates, particularly those containing methyl and nitro groups, suggesting a broader substrate scope. Additionally, we see that RufO binds to its redox partner CamB with micromolar affinity, and its interaction significantly enhances the putative substrate binding by ∼10-fold. Our crystal structure of RufO reveals similarities and differences in putative substrates and ferredoxin binding regions compared to other CYP450 enzymes. Our findings establish RufO might be a substrate-promiscuous enzyme with potential applications in the biocatalytic nitration of industrially relevant compounds.

Vitamin B and its vitamers are essential in bacteria. Many are auxotrophic for B vitamers and require salvage pathways and membrane uptake systems. Despite the importance of the uptake systems, very few transporters have been structurally characterized. The structure of the periplasmic binding protein (P5PA) of an ABC uptake transporter from the pathogen Actinobacillus pleuropneumoniae has been recently solved in complex with pyridoxal 5'-phosphate (PLP). Another close homolog from the same organism, AfuA, had been structurally characterized as a complex with glucose-6-phosphate (G6P). To study the molecular recognition of PLP by P5PA, a comparative approach has been applied. The heterologous complexes P5PA-G6P and AfuA-PLP have been generated by docking. Systematic molecular dynamics simulations have been applied to the native and heterologous complexes. Binding energies and molecular interactions have been compared for all the complexes. The results suggest the selective binding of the ligand is achieved by a combination of structural factors specific to each protein, including shape of the binding site, steric hindrance, hydrogen bonding, electrostatic and hydrophobic interactions. No single residue is uniquely responsible for ligand specificity although a few side chains play a significant role. Heterologous ligands are subject to destabilizing interactions that provoke the distortion of the ligand itself and the alteration of the protein dynamics. Residue Q267 appears to provide a significant stabilization contribution in the P5PA-PLP complex but not in AfuA-PLP. Likewise, D207 provides stabilization in the AfuA-G6P complex and not in AfuA-PLP. The indications obtained suggest strategies for the design of specific inhibitors.

The impact of solution conditions on the P isotropic chemical shifts of DNA phosphates and therefore the sequence-dependent backbone conformational equilibrium has not been well-documented. There are no previous studies of DNA backbone equilibrium in the presence of crowding agents, nor any on mismatched DNA. A systematic study of several experimental conditions (Na concentration, K concentration, Mg concentration, pH, the presence of PEG molecular crowders) was performed and the effect quantified in mismatched DNA compared to a canonical control sequence. Na concentration, pH and crowding agents have only a minimal effect on the backbone equilibrium (<5 % perturbation on backbone populations). But in the mismatched DNA, both K and Mg shift the backbone equilibrium on both DNA strands but most significantly perturb the phosphates in proximity to the mismatch. This indicates a possible role of counterions in mismatch recognition or nucleotide flipping, and suggests knowledge of solutions conditions continue to be relevant for conformational processes.

The interaction of protein with other biomolecules is central to all cellular processes. In particular, protein-lipid interactions play an essential role in regulating soluble and membrane protein function, structure, and dynamics. However, probing these interactions remains challenging due to the complexity and heterogeneity of membranes. Various methods have been developed to characterize protein-membrane interaction, each presenting advantages and limitations. This study presents a robust methodology based on continuous-wave Electron Paramagnetic Resonance (CW-EPR) spectroscopy to characterize protein-membrane interactions. We focused on the protein Tau, an intrinsically disordered protein associated with neurodegenerative diseases. We show that the interaction of labelled Tau with lipids gives rise to a very distinct lineshape, which can be used to quantify the fraction of bound protein. This allows to obtain the apparent binding mode and affinity through titration experiments. In addition, we show that a single measurement provides the absolute concentration of free and bound protein. We argue that this information, which is rarely obtained by other methods providing relative signals, is very useful for mechanistic studies. Furthermore, we developed a minimal-data approach and demonstrated that a single EPR measurement can be used to estimate an apparent binding constant. The approach is applied to the Tau-membrane interaction occurring in different conditions affecting the binding behavior. The presented methodology is expected to be applicable to other proteins.

Dopamine interaction with DNA has been studied using square-wave voltammetry (SWV), fluorescence, and absorption spectroscopy. The effect of dopamine on the binding of classical intercalating agents, such as acridine orange (AO), ethidium bromide (EtBr) as well as classical groove binding ligand Hoechst 33258 (H33258) was evaluated. The obtained results clarify the molecular mechanisms of dopamine interaction with DNA and reveal its potential competition for intercalation and minor groove binding sites which show that dopamine interacts with DNA by multimodal modes. On the basis of fluorescence measurements, the values of the binding constant (K) and the number of base pairs (n) per binding site were determined. It was revealed that the values of these binding parameters (K and n) depend on the ionic strength of the solution. Based on the changes of the binding parameters of AO, EtBr and H33258 with DNA in the absence and presence of dopamine, it was shown that the presence of the neurotransmitter reduces the affinity of these ligands toward DNA. The obtained data indicate that dopamine binds to DNA via intercalation and minor groove binding mechanisms, which leads to a decrease in the binding constants of these ligands with DNA. In the case of dopamine interaction with DNA in the presence of intercalators, the effect is especially pronounced for the strong (intercalation) binding mode of EtBr, as its binding constant with DNA in the presence of dopamine is significantly lower than that of AO.

Staphylococcus aureus (S. aureus) is a Gram-positive pathogenic bacterium and a major cause of nosocomial infections. Between 20 % and 50 % of S. aureus strains are resistant to a wide range of antibiotics. DMS-DA6-NH (DA6) is a novel antimicrobial peptide (AMP) that exhibits high efficacy against various bacterial strains, particularly S. aureus, by disrupting its membrane through an as-yet-unknown mechanism. We employed in vivoH solid state Nuclear Magnetic Resonance (NMR) to investigate the mode of action of AMPs on deuterated bacteria. This technique provides insights into membrane order and its changes with increasing AMP concentration. Our results enabled us to compare the mechanism of DA6 with those of AMPs with established modes of action. We found that DA6 induces pore formation in the membrane of S. aureus. This protocol serves as a template for determining the mechanisms of action of other peptides, an essential step for developing and patenting such drugs for the treatment of human diseases.

Recent work by Ben Abu and Wolfson introduces the concept of 'energetic memory' in ion channels, providing a mechanistic framework for ultrasound neuromodulation. This discussion examines how K2P (two-pore domain potassium) channels serve as primary mediators of mechanosensitive memory due to their small size (0.5 μm radius), constitutive activity, and critical physiological roles. In contrast, larger Kv channels (5 μm) show intermediate sensitivity while Na channels (50 μm) remain largely unaffected, creating size-dependent responses. Nanoindentor experiments demonstrate sustained membrane hyperpolarization following mechanical compression, validating the theoretical predictions. The energetic memory model explains ultrasound therapy's clinical efficacy through preferential K2P channel compression, energy system adaptation, and prolonged recovery phases. This framework enables rational optimization of therapeutic protocols and extends to other mechanically-based interventions, fundamentally expanding our understanding of neural plasticity beyond traditional electrical mechanisms.

Holy basil (Ocimum tenuiflorum) is primarily found in Nepal and India. In Ayurveda, it is commonly used as a traditional medicine to reduce pain, swelling, and various diseases. It has gained significant attention for its potential anti-inflammatory properties. One of the key targets associated with inflammation is Cyclooxygenase-2 (COX-2), an enzyme responsible for prostaglandin synthesis during the inflammatory response. In this study, we selected twenty flavonoids in the Holy Basil plant. These compounds were screened through Lipinski's Rule of Five, followed by ADMET prediction. Virtual screening was conducted on the selected compounds against the COX-2 enzyme as a receptor using molecular docking techniques. Molecular docking study provides valuable insights at the molecular level into the interactions between Holy Basil compounds and COX-2. Furthermore, density functional computations were carried out utilizing the B3LYP method with the 6-311G basis, which is set to gain insight into the structural and electronic properties of the compounds. This study showcases the potential of flavonoids such as rhamnetin, Luteolin and kaempferol to act as anti-inflammatory agents, sparking further interest and research in this area.

Nuclear receptors (NRs) are multidomain, ligand-activated transcription factors that play critical physiological roles. While the structured DNA-binding domain (DBD) and ligand-binding domain (LBD) have been well characterized, the function of intrinsically disordered regions-such as the hinge between the LBD and DBD-remains unclear. To illuminate the role of the hinge, we conducted five-microsecond molecular dynamics simulations of thyroid hormone receptor (TRα) alone versus bound to DNA. We reveal that DNA binding induces a significant structural change in the hinge region (helical to unwound coil), with a potentially important role in the regulation of TRα activity. Previously, hinge helicity has been reported to drive oligomerization and the consequent inhibition of coactivator binding, and such DNA-induced transition may promote TR activation. Protein-DNA binding is found to be multivalent and contains the direct interaction of the hinge with the DNA minor groove in addition to the canonical recognition helix of the DBD with the major groove. Furthermore, the poly-Arg segment of the hinge has a direct and significant influence on DNA conformation. This interaction promotes a bent DNA phosphate backbone, which might further contribute to the protein-DNA recognition. On a global scale, DNA binding induces a "closed-to-open" conformational change thus reducing direct DBD-LBD interactions, which corroborates previous calorimetric binding studies. Overall, our results provide insight into the mechanism of DNA recognition and the resulting conformational dynamics of the TRα-DNA complex.

Sodium dodecyl sulfate (SDS) is one of the most widely used detergents. Here, we discuss current knowledge regarding applications of SDS and its modes of interaction with proteins, particularly at low concentrations. SDS at 1-2 %, which is well above the critical micelle concentration, is commonly used to extract fully denatured and dissociated proteins and SDS polyacrylamide gel electrophoresis (SDS-PAGE) in various applications, especially proteomics. In contrast, low concentration SDS may have been relatively underutilized. Here, we demonstrate the use of 0.1 % SDS for decellularization and protein fractionation. Why is 0.1 % SDS unique? The interaction between SDS and proteins is complex and depends on both the conditions and the proteins involved. At 0.1 %, the effects of SDS appear to be intermediate between negligible and extensive binding, highlighting its potential for novel applications. Two milder anionic detergents, Sarkosyl and sodium N-lauroyglutamate, whose effects are similar in certain applications to those of low concentration SDS, were briefly discussed.

A comprehensive understanding of the molecular mechanism underlying the Liquid-Liquid Phase Separation (LLPS) pathway of LCD-TDP43 remains a challenge in the context of its neuropathogenesis. The primary driving force behind the TDP-43 LLPS is the interplay of hydrophobic interactions reinforced by aromatic residues. This study presents a novel, convenient, sensitive, and probe-free approach using excitation-emission matrix (EEM) fluorescence to monitor the microenvironment of aromatic residues and π-π stacking interactions during different stages of the LLPS pathway. Protein local structuring and the alterations in the positions of aromatic residues, individually and collectively, were detected by this life-time 3D fingerprinting. A new intermediate state with a unique α-sheet structure in the liquid droplet state and other transient species up to amyloid fibrils was discovered by CD and FTIR analyses. This structure with an inherent tendency for transition to β-amyloids, has not previously been reported in the context of LCD-TDP43 nor other LLPS-prone proteins. Mapping of hydrophobic clustering during phase separation revealed a continuous increase, accompanied by different surrounding polarities. The formation of distinct protein species within the LLPS pathway (from monomer to fibril), along with the amyloidogenic nature of TDP-43 fibrillation, was also confirmed by AFM analysis and ThT assay. To conclude, the 3D fluorescence method introduced in this study provides an effective and straightforward approach to critical valuable insights into the key π-π interactions in the LLPS-dependent aggregation pathway of LCD-containing IDPs. The novel identification of the α-sheet non-fibrilar intermediates may provide a new perspective for elucidating the aggregation mechanism of these proteins.

Silver nanoparticles (AgNPs) synthesized through green chemistry approaches offer a sustainable alternative to conventional methods, with potential applications in various biological fields. In this study, we report the synthesis of AgNPs using terpenoids derived from Ipomoea hederifolia L. (Convolvulaceae). The AgNPs (AgNPs-T) were characterized using UV-Vis spectroscopy, which revealed a surface plasmon resonance (SPR) peak at 452 nm, confirming successful synthesis. Fourier-transform infrared spectroscopy (FTIR) analysis identified functional groups such as hydroxyl and carbonyl that facilitated the reduction of silver ions and acted as stabilizing agents. Transmission electron microscopy (TEM) showed that the AgNPs-T were spherical in shape, with sizes ranging from 4 to 20 nm, and were well-dispersed due to the presence of capping agents from the plant extract. The biological activities of AgNPs-T were evaluated, showcasing potent antibacterial activity against several human pathogenic bacteria. Additionally, AgNPs-T exhibited significant antibiofilm and anti-quorum sensing activities, disrupting biofilm formation and inhibiting bacterial communication. The nanoparticles also demonstrated strong antioxidant properties by scavenging DPPH radicals in a dose-dependent manner. Moreover, cytotoxicity studies using the MTT assay revealed that AgNPs-T exerted dose-dependent anticancer effects against breast cancer (MCF-7) cells. These findings suggest that Ipomoea hederifolia-derived AgNPs possess multifunctional biological activities, making them promising candidates for applications in antimicrobial, antioxidant, and anticancer therapies.

There has been a growing concern regarding antibiotic resistance which has led to the design of new antibacterial nanocomposite with high efficiency and low cytotoxicity. This report demonstrates an approach for the design of a nanocomposite comprising of histidine tagged silver nanoparticles (His@AgNPs) embedded it in a carboxymethyl chitosan hydrogel (CHG). Carboxymethyl chitosan was used as a cationic polymer substrate to reinforce the antibacterial activity of the embedded silver nanoparticles. ICP-MS showed sufficient release of silver ions which results in enhanced antibacterial activity. In addition, the hydrogel with embedded silver nanoparticles exhibited excellent antibacterial performance against gram-negative Escherichia coli and gram-positive Staphylococcus aureus through a "kill-release" stratagem, which was mainly due to the synergistic effect of CHG and Ag ions release. The results showed that the cross-linking may enhance antibacterial activities of nanocomposite (CHG-His@AgNPs) by approximately 4-folds in comparison to His@AgNPs. A strong signal observed in confocal microscopy confirmed the successful internalization of the nanocomposite in tested microorganisms. The change in the morphology of the bacterial cells upon the treatment with nanocomposite was observed through FE-SEM microscopy which confirmed the antibacterial activity of the nanocomposite. The synthesized nanocomposite can thereby serve as an excellent candidate for tackling antibiotic resistance.

The rubber antioxidant, N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6-PPD), as an emerging pollutant, is receiving more and more attention recently. This study investigated the intermolecular interactions of 6-PPD with two key biological macromolecules, human serum albumin (HSA) and alpha-glucosidase (AG), to understand the transport and toxic effects of 6-PPD. Using multiple spectroscopic methods and molecular docking technology, the results demonstrated that 6-PPD could bind to both HSA and AG, thereby inducing fluorescence quenching and conformational changes in both proteins. The binding constants were determined to be (5.93 ± 0.20) × 10 and (3.17 ± 0.15) × 10 L mol respectively for HSA-6-PPD and AG-6-PPD systems at 298 K, revealing strong binding affinities. Molecular docking identified specific binding sites and non-covalent interactions of the two systems. MD and Energy decomposition analysis revealed the dynamics conformational changes of the complexes and identified van der Waals and electrostatic interactions as primary binding drivers for both systems, while polar solvation energy impeded complex formation. TYR161, ILE142, and TYR138 dominated HSA-6-PPD stabilization, whereas AG-6-PPD was driven by hydrophobic interactions with TRP1369 and VAL1373, with ARG1377 incurring substantial desolvation penalties. Synchronous fluorescence and circular dichroism spectroscopy indicated that 6-PPD binding did not disrupt the microenvironment of Tyr and Trp residues in HSA and AG, while induced structural alterations in HSA and AG that could affect their physiological function. In-vitro tests showed that 6-PPD inhibited AG activity in a dose-dependent manner, with an IC of 8.22 ± 0.44 μmol L. ADMET and PASS online tools was used to predict physicochemical properties and multiorgan toxicity. This work provided insights into the transport and molecular toxicity of 6-PPD, highlighting the adverse biological effects associated with this common rubber additive.

Cationic peptides have emerged as promising candidates in anticancer therapy due to their ability to directly target the plasma membrane of cancer cells, a mechanism that could potentially bypass traditional drug resistance pathways. In this study, we evaluated the cytotoxic activity and cell-membrane binding properties of three amphiphilic cationic peptides from the G(IIKK)ₙI-NH₂ family (n = 2-4) against human melanoma cells (SK-MEL-28). By performing MTT assays and tracking the propidium iodide (PI) uptake throughout peptide-cell interaction, we evaluated peptides' cytotoxicity. Assessment of the interaction dynamics was conducted by fluorescence spectroscopy assays with FPE, a surface potential sensitive probe. This evaluation indicated that an increase in net positive charge was relatable to a lower dissociation constant (K). Notably, G(IIKK)₄I-NH₂ showed the highest cytotoxicity, significant morphological alterations, rapid membrane permeabilization, and the lowest K, indicating a stronger membrane affinity when compared to the other peptides. G(IIKK)I-NH₂, in the same manner as G(IIKK)₄I-NH₂, revealed a cooperative binding behavior, evidenced by a Hill coefficient > 1. An inverse correlation between peptide-cell membrane dissociation constants and cytotoxicity was established, supporting the notion that membrane interaction is a critical determinant of anticancer activity. In addition, we used a cell surface membrane potential probe to possibly anticipate the in vitro activity of cationic peptides. Altogether, these findings provide mechanistic insights into peptide-cell membrane interactions and may offer a basis for the rational design of novel anticancer peptides targeting melanoma.

Diabetes and the related comorbidities have been associated with elevated levels of advanced glycation end products (AGEs). The biochemical process of advanced glycation, is believed to be playing a pivotal role in the development of complications. Since there exists a great deal of promise for natural products offering antidiabetic potential, we studied advanced glycation inhibition and anti-diabetic profile of Phytolacca latbenia (Moq). on fractions and the sorted compounds based on the QSAR and molecular docking analysis. The top two bioactive compounds; Kaempferol and Esculentoside G, were further evaluated for the MD simulation studies at 150 ns run, compared with the standard. Among the tested compounds, Kaempferol presented significant binding energies in MM-GBSA (-48.63 Kcal/mol) and MD simulation studies (73 %) with transcriptional regulator 4F5S. Molecular docking studies revealed that kaempferol formed three hydrogen bonds with Val342, Ser343 and Ser453, along with Pi-Pi stacking and Pi-cation interactions with Trp213 and Arg217 residues of the 4F5S protein. Kaempferol also displayed significant α-glucosidase inhibition (IC 0.042 ± 2.31 μg/ml) compared to the acarbose (IC 0.036 ± 0.31 μg/ml). Almost all of the selected compounds demonstrated adherence to the safety requirements established by ADMET investigation. Liquid-liquid partitioning of the crude methanolic extract with solvents of increasing polarity yielded five solvent fractions;the ethyl acetate fraction (ETOA) obtained by liquid-liquid partitioning of the crude extract with ethyl acetate and water proclaimed substantial results in both the non-oxidative (61 %) and oxidative (58 %) antiglycation assays for thiol group estimation. The ethylacetae fraction (ETOA) demonstrated comparatively strong antioxidant activity, with an IC₅₀ value of 13.25 ± 0.69 μg/ml as determined by the DPPH assay. In α-glucosidase assay, Aqueous fraction demonstrated a considerable inhibition with IC value of 0.108 ± 0.32 μg /ml compared to the standard (IC 0.083 ± 0.43 μg/ml). The safety assessment revealed a slight decline in HeLa cell viability, dropping from 82 % at a 2.5 % concentration to 69 % at a 10 % concentration over 24 h, relative to the control.Therefore, Phytolacca latbenia (Moq). and its phytocompounds tested inhibit α-glucosidase and Advanced glycation end product-the process that underlie diabetic complications and may therefore holds great promise as therapeutic agent, with no toxicity concern,against diabetes and related comorbidities.

Caveolin-1 (Cav1) is an integral membrane protein essential for the formation of caveolae, plasma microdomains implicated in signal transduction and mechanoprotection. Cav1 is comprised of three major alpha helices, but the topology these helices adopt remains unclear. Proline 110 is located between helix 1 and helix 2, and is hypothesized to enable Cav1 to adopt an intramembrane turn crucial for the cytosolic topology of Cav1. To assess the structural role of Proline 110, we utilized Förster resonance energy transfer (FRET) between native tryptophan (W128) and site-specifically labeled dansyl fluorophores to monitor conformational changes induced by the mutation of Proline 110 to Alanine (P110A). Static light scattering confirmed that all FRET constructs behaved monomerically, ensuring intramolecular energy transfer measurements. Our results show a significant decrease in FRET efficiency upon the P110A mutation consistent with a large conformational change. These findings support the critical role of P110 in maintaining the native topology of Cav1 and highlights the structural sensitivity of the intramembrane turn.

Bovine Serum Albumin (BSA) is a globular, water-soluble protein widely used as a model system due to its stability, binding capacity, and structural similarity to human serum albumin (HSA). Cold atmospheric plasma (CAP) has emerged as a versatile tool for biomolecule modification, sterilization, food preservation, and wound healing. This study explores the effects of CAP on glycated BSA, focusing on structural and self-assembly processes. SEM analysis reveals that CAP induces distinct protein self-assemblies depending on treatment duration. Thioflavin assays show increased fluorescence intensity in CAP-treated glycated BSA compared to native and glycated BSA, indicating an enhancement in β-sheet content and self-assembly. These findings offer valuable insights into CAP's role in modulating protein structures, with implications for biomaterials, disease mechanisms, and protein engineering.

AFM-IR combinates atomic force microscopy and infrared spectroscopy to compensate the limitations of both techniques taken separately. It has been reviewed for a large application field like polymers, geology and life sciences. In biology, it is an important tool to study amyloids and protein aggregation processes. Indeed, misfolding can appear under various circumstances in the process of globular proteins folding. In the case of amyloidosis, fibrillar aggregates are deposited in intracellular inclusions or in tissues as extracellular plaques. These aggregates (oligomers or fibrils) are characterized by high β-sheet content which can be analyzed in AFM-IR thanks to specific absorption band. The main progresses and developments of this technique are summarized since its creation in 2005. The evolution of laser sources and new measurement modes has led to the development of new instruments. They are always more efficient, allowing faster analysis, a wider sample range or more sensitive in order to give more (chemical) information about the sample. An overview of the progress made in photothermal AFM-IR in characterization of amyloids and amyloid aggregation processes is also described. The tapping and resonance-enhanced contact AFM-IR are the most commonly used modes. Generally, the label-free analysis of the conformation of the oligomers and/or fibrils at micromolar concentration is described, either in an aggregation kinetic study or in analysis of fibrils in ex vivo study. The coaggregation of two amyloids is also realized using C-labeled peptide to distinguish both two spectral signatures.

This study reports a microwave-assisted green synthesis of cobalt and gadolinium dual doped ZnO nanoparticles using Phyllanthus emblica extract as a natural bio-stabilizer. The plant-derived phytochemicals enabled an eco-friendly process and improved nanoparticle stability. Structural, morphological, and optical analyses (TEM, XRD, SEM, EDX, UV-Vis) confirmed the successful formation of uniformly dispersed nanoparticles. The crystallite size obtained from XRD decreased from 26.59 to 20.59 nm with increasing Gd content, while TEM analysis showed slightly larger particle sizes ranging from 28.39 to 23 nm, validating the nanoscale dimensions and doping-induced size reduction. The (Co, Gd) dual doped ZnO nanoparticles exhibited strong antibacterial and antioxidant activity, demonstrating their potential to mitigate oxidative stress. Their photocatalytic efficiency was further confirmed through up to 97% degradation of Methyl Orange and Methylene Blue dyes. These combined results demonstrate that green synthesis and dual doping synergistically enhance the functional properties of ZnO nanoparticles, positioning them as promising candidates for environmental remediation, healthcare applications, sunscreen formulations, and active food packaging systems.