Search Results (19)

Excessive copper ions in the human body can cause a variety of diseases, such as gastrointestinal disorders, cirrhosis, and Alzheimer’s disease. Techniques like Inductively Coupled Plasma–Mass Spectroscopy and Atomic Absorption Spectroscopy are available for copper detection, but the associated cost issues for sample preparation and labor limit their application for on-site detection. Herein, we are reporting a versatile method for detecting copper ions using a peptide-functionalized gold nanoparticle sensor in combination with various optical spectroscopic techniques. The peptide (CW) exhibited selective sensing ability for Cu(II) with visual colorimetric and optical spectroscopic changes compared to other metal ions tested. CW showed a visual colorimetric response from colorless to light brown color after interaction with Cu(II). Converting CW to a gold nanoparticle appended (CW-AuNPs) nanoplatform enabled a multimodal detection platform for Cu (II), which utilizes colorimetric and optical spectrum changes and surface-enhanced Raman spectroscopy (SERS) to enable highly sensitive sensing of Cu(II), even at extremely low concentrations (76 nms.). CW-AuNPs exhibit a controlled aggregation property in the presence of Cu(II), resulting in the creation of hot spots for SERS-based detection. Moreover, the peptide unit attached to the gold nanoparticles serves both as a binding motif for Cu(II) and as a Raman reporter for Cu(II) sensing. Our comprehensive analysis, including solution-state and dry-mapping Raman spectroscopic studies, demonstrates remarkable picomolar sensitivity of the peptide–gold nanoparticle system for Cu(II) detection. Moreover, we prepared a paper test strip from CW-AuNPs and used it as a visual colorimetric platform for sensitive detection of copper ions. Full article

Studying membrane proteins in a native environment is crucial to understanding their structural and/or functional studies. Often, widely accepted mimetic systems have limitations that prevent the study of some membrane proteins. Micelles, bicelles, and liposomes are common biomimetic systems but have problems with membrane compatibility, limited lipid composition, and heterogeneity. To overcome these limitations, polymersomes and hybrid vesicles have become popular alternatives. Polymersomes form from amphiphilic triblock or diblock copolymers and are considered more robust than liposomes. Hybrid vesicles are a combination of lipids and block copolymers that form vesicles composed of a mixture of the two. These hybrid vesicles are appealing because they have the native lipid environment of bilayers but also the stability and customizability of polymersomes. Gramicidin A was incorporated into these polymersomes and characterized using continuous wave electron paramagnetic resonance (CW-EPR) and transmission electron microscopy (TEM). EPR spectroscopy is a powerful biophysical technique used to study the structure and dynamic properties of membrane proteins in their native environment. Spectroscopic studies of gramicidin A have been limited to liposomes; in this study, the membrane peptide is studied in both polymersomes and hybrid vesicles using CW-EPR spectroscopy. Lineshape analysis of spin-labeled gramicidin A revealed linewidth broadening, suggesting that the thicker polymersome membranes restrict the motion of the spin label more when compared to liposome membranes. Statement of Significance: Understanding membrane proteins’ structures and functions is critical in the study of many diseases. In order to study them in a native environment, membrane mimetics must be developed that can be suitable for obtaining superior biophysical data quality to characterize structural dynamics while maintaining their native functions and structures. Many currently widely accepted methods have limitations, such as a loss of native structure and function, heterogeneous vesicle formation, restricted lipid types for the vesicle formation for many proteins, and experimental artifacts, which leaves rooms for the development of new biomembrane mimetics. The triblock and diblock polymersomes and hybrid versicles utilized in this study may overcome these limitations and provide the stability and customizability of polymersomes, keeping the biocompatibility and functionality of liposomes for EPR studies of membrane proteins. Full article

Background: Protein hydrolysates from insects are recognized for their biological activities. Black soldier fly larvae (BSFL) have drawn attention due to their antioxidant protein hydrolysates. However, research on bioactive peptides derived from these hydrolysates, particularly their cancer chemopreventive potential, remains limited. This study aims to evaluate the antioxidant, anti-inflammatory, antimutagenic, and anticancer activities of BSFL-derived bioactive peptides and explore the molecular mechanisms. Methods: Alkali-soluble BSFL protein (ASBP) was extracted and hydrolyzed using Alcalase and bromelain under optimized conditions. Antioxidant activity was assessed via FRAP, ABTS, and DPPH assays. The hydrolysate with the highest antioxidant activity was fractionated into molecular weight (MW) groups (>30, 10, and <3 kDa). The bioactivity of fractionated peptides was evaluated through antioxidant, anti-inflammatory (nitric oxide production in RAW 264.7 cells), antimutagenic (Ames test), and anticancer (CCK-8 assay on HCT 116, COLO205, Cw-2, and Caco-2 cells) assays. Mechanistic insights were obtained via microarray and Western blot analyses. Peptides were identified by LC-MS/MS. Results: The ASBP-Alcalase hydrolysate (ASBP-AH) showed optimal antioxidant activity at 3% (w/w) for 4 h. The ASBP-AH 30 (MW > 30 kDa) fraction exhibited the highest antioxidant capacity. In contrast, the ASBP-AH3 (MW < 3 kDa) fraction exhibited significant antimutagenic effects, reduced nitric oxide production, and decreased COLO205 cell viability. Treatment with ASBP-AH3 at its LC₅₀ dose modulated the SKP2/p21/cyclin D1 pathways. Mostly peptides from ASBP-AH3 were composed of hydrophobic and charged amino acids. Conclusions: BSFL-derived bioactive peptides exhibit potential as multifunctional agents for cancer chemoprevention. In vivo studies are required to explore their clinical applications. Full article

Bgl2p is a major, conservative, constitutive glucanosyltransglycosylase of the yeast cell wall (CW) with amyloid amino acid sequences, strongly non-covalently anchored in CW, but is able to leave it. In the environment, Bgl2p can form fibrils and/or participate in biofilm formation. Despite a long study, the question of how Bgl2p is anchored in CW remains unclear. Earlier, it was demonstrated that Bgl2p lost the ability to attach in CW and to fibrillate after the deletion of nine amino acids in its C-terminal region (CTR). Here, we demonstrated that a Bgl2p anchoring is weakened by substitution Glu-233/Ala in the active center. Using AlphaFold and molecular modeling approach, we demonstrated the role of CTR on Bgl2p attachment and supposed the conformational possibilities determined by the presence or absence of an intramolecular disulfide bond, forming by Cys-310, leading to accessibility of amyloid sequence and β-turns localized in CTR of Bgl2p for protein interactions. We hypothesized the mode of Bgl2p attachment in CW. Using atomic force microscopy, we investigated fibrillar structures formed by peptide V187MANAFSYWQ196 and suggested that it can serve as a factor leading to the induction of amyloid formation during interaction of Bgl2p with other proteins and is of medical interest being located close to the surface of the molecule. Full article

Mackerel (Auxis thazard), a tropical dark-fleshed fish, is currently a viable resource for the manufacture of surimi, but the optimal washing procedure for more efficient use of this particular species is required right away. Washing is the most critical stage in surimi production to ensure optimal gelation with odorless and colorless surimi. The goal of this study was to set a simple washing medium to the test for making mackerel surimi. Washing was performed three times with different media. T1 was washed with three cycles of cold carbonated water (CW). T2, T3, and T4 were washed once with cold CW containing 0.3%, 0.6%, or 0.9% NaCl, followed by two cycles of cold water. T5, T6, and T7 were produced for three cycles with CW containing 0.3%, 0.6%, or 0.9% NaCl. For comparison, unwashed mince (U) and conventional surimi washed three times in cold tap water (C) were employed. The maximum yield (62.27%) was obtained by washing with T1. When varying quantities of NaCl were mixed into the first washing medium (T2–T4), the yield decreased with increasing NaCl content (27.24–54.77%). When washing with NaCl for three cycles (T5–T7), the yield was greatly decreased (16.69–35.23%). Conventional surimi washing (C) produced a yield of roughly 40%, which was comparable to T3. Based on the results, treatments that produced lower yields than C were eliminated in order to maximize the use of fish resources and for commercial reasons. The maximum NaCl content in CW can be set at 0.6% only during the first washing cycle (T3). Because of the onset of optimal unfolding as reported by specific biochemical characteristics such as Ca²⁺-ATPase activity (0.2 μmol inorganic phosphate/mg protein/min), reactive sulfhydryl group (3.61 mol/10⁸ g protein), and hydrophobicity (64.02 µg of bromophenol blue bound), T3 washing resulted in surimi with the greatest gel strength (965 g.mm) and water holding capacity (~65%), with fine network structure visualized by scanning electron microscope. It also efficiently removed lipid (~80% reduction), myoglobin (~65% reduction), non-heme iron (~94% reduction), and trichloroacetic acid-soluble peptide (~52% reduction) contents, which improves whiteness (~45% improvement), reduces lipid oxidation (TBARS value < 0.5 mg malondialdehyde equivalent/kg), and decreases the intensity of the gel’s fishy odor (~30% reduction). As a result, washing mackerel surimi (A. thazard) with CW containing 0.6% (w/v) NaCl in the first cycle, followed by two cycles of cold water washing (T3), can be a simple method for increasing gel-forming capability and oxidative stability. The mackerel surimi produced using this washing approach has a higher quality than that produced with regular washing. This straightforward method will enable the sustainable use of dark-fleshed fish for the production of surimi. Full article

Wound healing is a complex process involving blood cells, extracellular matrix, and parenchymal cells. Research on biomimetics in amphibian skin has identified the CW49 peptide from Odorrana grahami, which has been demonstrated to promote wound regeneration. Additionally, lavender essential oil exhibits anti-inflammatory and antibacterial activities. Given these considerations, we propose an innovative emulsion that combines the CW49 peptide with lavender oil. This novel formulation could serve as a potent topical treatment, potentially fostering the regeneration of damaged tissues and providing robust antibacterial protection for skin wounds. This study investigates the physicochemical properties, biocompatibility, and in vitro regenerative capacity of the active components and the emulsion. The results show that the emulsion possesses appropriate rheological characteristics for topical application. Both the CW49 peptide and lavender oil exhibit high viability in human keratinocytes, indicating their biocompatibility. The emulsion induces hemolysis and platelet aggregation, an expected behavior for such topical treatments. Furthermore, the lavender-oil emulsion demonstrates antibacterial activity against both Gram-positive and Gram-negative bacterial strains. Finally, the regenerative potential of the emulsion and its active components is confirmed in a 2D wound model using human keratinocytes. In conclusion, the formulated emulsion, which combines the CW49 peptide and lavender oil, shows great promise as a topical treatment for wound healing. Further research is needed to validate these findings in more advanced in vitro models and in vivo settings, potentially leading to improved wound-care management and novel therapeutic options for patients with skin injuries. Full article

The genetic variants of HLAs (human leukocyte antigens) play a crucial role in the virus–host interaction and pathology of COVID-19. The genetic variants of HLAs not only influence T cell immune responses but also B cell immune responses by presenting a variety of peptide fragments of invading pathogens. Peptide cocktail vaccines produced by using various conserved HLA-A2 epitopes provoke substantial specific CD8+ T cell responses in experimental animals. The HLA profiles vary among individuals and trigger different T cell-mediated immune responses in COVID-19 infections. Those with HLA-C*01 and HLA-B*44 are highly susceptible to the disease. However, HLA-A*02:01, HLA-DR*03:01, and HLA-Cw*15:02 alleles show resistance to SARS infection. Understanding the genetic association of HLA with COVID-19 susceptibility and severity is important because it can help in studying the transmission of COVID-19 and its physiopathogenesis. The HLA-C*01 and B*44 allele pathways can be studied to gain insight into disease transmission and physiopathogenesis. Therefore, integrating HLA testing is suggested in the ongoing pandemic, which will help in the rapid identification of highly susceptible populations worldwide and possibly acclimate vaccine development. Therefore, understanding the correlation between HLA and SARS-CoV-2 is critical in opening new insights into COVID-19 therapeutics, based on previous studies conducted. Full article

Wound healing in adult mammals results in scar formation, which prevents recovering the full functionality of the original skin. This area of dermatology is constantly evolving and especially focuses on aging and the design of recovery treatments for skin burns. Because it has been reported that Lavandula angustifolia essential oil and the CW49 peptide present regenerative, healing, and anti-inflammatory effects, we selected these two natural compounds to formulate a topical treatment with potential regenerative capability. This was accomplished by synthesizing oil-in-water (O/W) emulsions at 10:90% w/w with lavender oil and the CW49 peptide. The formulations were characterized physicochemically and evaluated in terms of biocompatibility, antibacterial activity, and wound-healing potential. The results showed that emulsions exhibited a droplet size of about 1 µm, a marked pseudoplastic behavior and a superior shelf stability of over 9 months. Additionally, they induced 35% hemolysis when compared with the positive control (similar to commercially available controls), induced platelet aggregation, and have a potent antibacterial activity against Staphylococcus aureus (20% of growth inhibition). The wound-healing potential was preliminarily evaluated for the CW49 peptide in a 2D scratch wound model of human keratinocytes, demonstrating an effective concentration for closure of 20 μg/mL. Thus far, we established a pipeline to develop and characterize the regenerative potential of bio-based topical treatments, particularly those based on lavender essential oil and the CW49 peptide. Full article

Detecting residual nasopharyngeal carcinoma (rNPC) can be difficult because of the coexistence of occult tumours and post-chemoradiation changes, which poses a challenge for both radiologists and surgeons using current imaging methods. Currently, molecular imaging that precisely targets and visualises particular biomarkers in tumours may exceed the specificity and sensitivity of traditional imaging techniques, providing the potential to distinguish tumours from non-neoplastic lesions. Here, we synthesised a HER2/SR-BI-targeted tracer to efficiently position NPC and guide surgery in living mice. This bispecific tracer contained the following two parts: IRDye 800 CW, as an imaging reagent for both optical and optoacoustic imaging, and a fusion peptide (FY-35), as the targeting reagent. Both in vitro and in vivo tests demonstrated that the tracer had higher accumulation and longer retention (up to 48 h) in tumours than a single-targeted probe, and realised sensitive detection of tumours with a minimum size of 3.9 mm. By visualising the vascular network via a customised handheld optoacoustic scan, our intraoperative fluorescence molecular imaging system provides accurate guidance for intraoperative tumour resection. Integrating the advantages of both optical and optoacoustic scanning in an intraoperative image-guided system, this method holds promise for depicting rNPC and guiding salvage surgery. Full article

The progressive accumulation of misfolded α-synuclein (α-syn) in the brain is widely considered to be causal for the debilitating clinical manifestations of synucleinopathies including, most notably, Parkinson’s disease (PD). Immunotherapies, both active and passive, against α-syn have been developed and are promising novel treatment strategies for such disorders. To increase the potency and specificity of PD vaccination, we created the ‘Win the Skin Immune System Trick’ (WISIT) vaccine platform designed to target skin-resident dendritic cells, inducing superior B and T cell responses. Of the six tested WISIT candidates, all elicited higher immune responses compared to conventional, aluminum adjuvanted peptide-carrier conjugate PD vaccines, in BALB/c mice. WISIT-induced antibodies displayed higher selectivity for α-syn aggregates than those induced by conventional vaccines. Additionally, antibodies induced by two selected candidates were shown to inhibit α-syn aggregation in a dose-dependent manner in vitro. To determine if α-syn fibril formation could also be inhibited in vivo, WISIT candidate type 1 (CW-type 1) was tested in an established synucleinopathy seeding model and demonstrated reduced propagation of synucleinopathy in vivo. Our studies provide proof-of-concept for the efficacy of the WISIT vaccine technology platform and support further preclinical and clinical development of this vaccine candidate. Full article

This study aimed to compare ozone-microbubble-washing (OM) performed by domestic equipment with conventional water-washing (CW) regarding resultant quality attributes of muscle foods. For this purpose, muscle microstructure and lipid and protein oxidation were evaluated in pork and fish samples after OM and CW treatments. The assessment of muscle microstructure showed that OM treatment did not damage the microstructure of muscle fibers in both pork and fish samples. Thiobarbituric acid reactive substances (TBARS) values were not detected in both treatment groups, and they were substantially below the generally acceptable threshold (1 mg MDA/kg). The methylglyoxal (MGO) level of OM-treated fish samples was significantly higher than that of CW-treated fish samples. However, glyoxal (GO) and MGO levels of OM-treated pork samples were significantly lower than that of CW-treated pork samples. Similar types and sites of oxidative modification and similar numbers of modified peptides, as well as no significant difference in the concentration of total and most of the free amino acids (FAA) between treatment groups, indicated that OM treatment did not accelerate protein oxidation. Full article

Disordered molecular solids present a rather broad class of substances of different origin—amorphous polymers, materials for photonics and optoelectronics, amorphous pharmaceutics, simple molecular glass formers, and others. Frozen biological media in many respects also may be referred to this class. Theoretical description of dynamics and structure of disordered solids still does not exist, and only some phenomenological models can be developed to explain results of particular experiments. Among different experimental approaches, electron paramagnetic resonance (EPR) applied to spin probes and labels also can deliver useful information. EPR allows probing small-angle orientational molecular motions (molecular librations), which intrinsically are inherent to all molecular solids. EPR is employed in its conventional continuous wave (CW) and pulsed—electron spin echo (ESE)—versions. CW EPR spectra are sensitive to dynamical librations of molecules while ESE probes stochastic molecular librations. In this review, different manifestations of small-angle motions in EPR of spin probes and labels are discussed. It is shown that CW-EPR-detected dynamical librations provide information on dynamical transition in these media, similar to that explored with neutron scattering, and ESE-detected stochastic librations allow elucidating some features of nanoscale molecular packing. The possible EPR applications are analyzed for gel-phase lipid bilayers, for biological membranes interacting with proteins, peptides and cryoprotectants, for supercooled ionic liquids (ILs) and supercooled deep eutectic solvents (DESs), for globular proteins and intrinsically disordered proteins (IDPs), and for some other molecular solids. Full article

Ferulic acid (FA) is known for its excellent antioxidant properties, which can provide many health benefits. One of its drawbacks is its instability under UVA light, which limits its potency. In this study, the new peptides LW2 (QNKRFYFRKNQ) and CW2 (a cyclic form of LW2) were designed based on bovine serum albumin site IIA conformation. A UVA irradiation experiment was performed to investigate the protective ability of these peptides towards FA against UVA damage. The percentages of FA remaining under UV irradiation due to the protection of CW2 and LW2 were 83% and 76%, respectively. The results showed the importance of the cationic residues and hydrophobic residues included in the peptide sequences. Moreover, the cyclic rigid structure showed greater protective ability as compared to its linear counterpart. Full article

The cell wall (CW) is a dynamic structure extensively remodeled during plant growth and under stress conditions, however little is known about its roles during the immune system priming, especially in crops. In order to shed light on such a process, we used the Phaseolus vulgaris-Pseudomonas syringae (Pph) pathosystem and the immune priming capacity of 2,6-dichloroisonicotinic acid (INA). In the first instance we confirmed that INA-pretreated plants were more resistant to Pph, which was in line with the enhanced production of H₂O₂ of the primed plants after elicitation with the peptide flg22. Thereafter, CWs from plants subjected to the different treatments (non- or Pph-inoculated on non- or INA-pretreated plants) were isolated to study their composition and properties. As a result, the Pph inoculation modified the bean CW to some extent, mostly the pectic component, but the CW was as vulnerable to enzymatic hydrolysis as in the case of non-inoculated plants. By contrast, the INA priming triggered a pronounced CW remodeling, both on the cellulosic and non-cellulosic polysaccharides, and CW proteins, which resulted in a CW that was more resistant to enzymatic hydrolysis. In conclusion, the increased bean resistance against Pph produced by INA priming can be explained, at least partially, by a drastic CW remodeling. Full article

Wetlands and aquatic ecosystems, which are an important part of the ecological system and national resources that need to be well managed, are becoming polluted by toxic heavy metals (HMs) from the industrial mining and smelting of metalliferous ores, and agricultural activities. The loss of wetlands may cause a loss of flora and fauna and thereby decrease biodiversity. Waterlogging resistant plants, their root associated microbes (Arbuscular Mycorrhizal Fungi (AMF)) and plant growth promoting rhizobia (PGPR) can provide potential tools in constructed wetlands (CWs) in order to allow for the Nano-Mycorrhizo-Phytoremediation (NMPR) of HM-polluted natural wetlands and aquatic ecosystems. AMF-CW systems should be considered ideal locations for the technical installations for phytoremediation and need to be optimized for the efficient functioning of phytoremediation in field trials. This presentation will address one of the major hurdles in the production of large quantities of indigenous and stress-adapted AMF inoculum for the purposes of constructing artificial AM-CW systems. The significance and potential role of floating islands of aquatic macrophytes such as Vetiver grass and their root associated microbes (AMF and PGPR) in an environmental cleanup of HM contaminated industrial, municipal, and mining effluents, will be highlighted in the presentation. During the environmental and pollutant stresses, the aquatic macrophytes and their root associated microbes produce nano-molecules of HM-binding cysteine-rich peptides, phytochelators (nano-molecules) forming HM-complexes that sequester HM-ions, protecting the host from contaminants. HM-adapted AMF not only enhance Vetiver grass growth, producing a greater biomass for bio energy production, but also uptake/stabilize HMs, e.g., Nano-Mycorrhizo-Phytoremediation (NMPR). Full article