Search Results (21)

Background: Rapid on-site evaluation (ROSE) is crucial for improving the diagnostic yield of fine-needle aspiration (FNA) biopsies. Despite recent advances in ROSE, such as telecytology, the rapid stains used in this process have not seen significant innovation. Diff-Quik (DQ) and Toluidine Blue (TB), the most common ROSE stains, have significant limitations. This study evaluates the optimized Toluidine Blue stain, a mixture of Toluidine Blue, Eosin, and Alcohol (TEA), as a potential alternative to TB or DQ for ROSE. Methods: A comparative study was conducted using fifty remnant body fluid specimens with adequate cellularity, collected at the University of California Davis Medical Center over six months. Two smears were prepared from each specimen. One was stained with TB, and the other with optimized Toluidine Blue (TEA). Digital images of each slide were evaluated by three cytologists and two cytopathologists, blinded to the stain, using five criteria: background staining, cytoplasmic detail, nuclear membrane clarity, chromatin texture, and nucleoli staining. Each criterion was scored on a scale of 1 to 3. Results: Optimized Toluidine Blue (TEA) stain demonstrated superior overall image quality compared to TB. Specifically, optimized Toluidine Blue (TEA) showed significantly less background staining (p < 0.05) and improved nuclear membrane clarity (p < 0.05), chromatin texture (p < 0.05), and nucleoli detail (p < 0.05). There was no significant difference between the two stains in the assessment of cellularity or cytoplasmic detail. Conclusions: The optimized Toluidine Blue (TEA) stain shows promise as a rapid stain for ROSE, offering rapid processing and improved digital image quality. Further evaluation of optimized Toluidine Blue (TEA) stain on FNA specimens is warranted to validate these findings and explore its potential to enhance telecytology in ROSE. Full article

The complexes of negatively charged polysaccharides with lipid vesicles have been shown to have applications in medicine, bioremediation, water purification, and construction of nano-biosensors. This article presents research on the formation of these complexes based on the interactions between three types of liposomes, DOPC liposomes (which contain a lipid bilayer in the liquid-disordered (Ld) state), RAFT liposomes (which contain liquid-ordered (Lo) lipid raft domains surrounded by lipids in the Ld state) and SPH–CHL liposomes (which contain a lipid bilayer in the Lo state), and two selected anionic polysaccharides, polysialic acid (PSA) and polygalacturonic acid (PGA). The analysis was conducted using a toluidine blue (TB) probe and the absorption spectroscopy technique. In contrast to DOPC and SPH–CHL liposomes, binding of negatively charged PSA or PGA chains to RAFT liposomes induced a TB absorption maximum shift from 630 nm to 560 nm. The obtained results indicate that toluidine blue can be applied for monitoring the formation of these nano-complexes, and that the boundaries between Ld/Lo domains within membranes in RAFT liposomes can significantly enhance the binding affinity of negatively charged polysaccharides to the lipid bilayer surface. The observed metachromatic shift in TB absorption suggests that negatively charged PSA and PGA chains interact with the Ld/Lo boundaries within RAFT liposome membranes. Full article

Cassava is a tropical tuberous root crop, feeding over a billion people globally. However, research on the chemical composition and bioactive effects of cassava leaves remains scarce. Two specific varieties of South China No. 9 (green leaves (G.L.)) and South China No. 20 (purple leaves (P.L.)) were investigated in this study. The components of G.L. and P.L. were analyzed under different extraction methods using ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Results showed that cassava leaf extracts are rich in bioactive metabolites such as D-(+)-mannose, trigonelline, rutin, kaempferol-3-O-rutinoside, and oleamide. To assess the anti-inflammatory efficacy of bioactive compounds, animal models were established. Compared to the histamine group (NA), the group treated with the extracts had reduced epidermal thickness in hematoxylin and eosin (HE) staining. Further analysis revealed a drastic reduction in the number of mast cells in toluidine blue (TB) staining and expression levels of inflammatory cytokines (IL-17 and TNF-α) in immunohistochemistry (IHC) staining. The ethanolic extracts from the leaves demonstrated potent anti-inflammatory activities, with the extract from G.L. surpassing that from P.L. Transcriptomic analyses propose that the anti-inflammatory effects of cassava leaves may be related to the modulation of genes involved in mast cell activation, such as Cma1, Cpa3, and Fn1, among others. Network pharmacology unveiled that the extract of cassava leaves modulates pathways associated with apoptosis, inflammation, and metabolism. Molecular docking revealed strong binding interactions between 1-stearoylglycerol and oleamide from cassava leaves extracts and the proteins of AKT1, TNF, and BRAF. Overall, cassava leaf extracts seem to be a promising natural anti-inflammatory agent. Full article

Detecting multiple tumor markers is of great importance. It helps in early cancer detection, accurate diagnosis, and monitoring treatment. In this work, gold nanoparticles–toluidine blue–graphene oxide (AuNPs-TB–GO) and gold nanoparticles–carboxyl ferrocene–tungsten disulfide (AuNPs–FMC–WS₂) nanocomposites were prepared for labeling Carcinoembryonic antigen (CEA) antibody and Carbohydrate antigen 72–4 (CA72-4) antibody, respectively, and used as two kinds of probes with different electrochemical signals. With the excellent magnetic performance of biotin immune magnetic beads (IMBs), the biofunctional IMBs were firmly deposited on the magnetic glassy carbon electrode (MGCE) surface by applying a constant magnetic field, and then the CEA and CA72-4 antibody were immobilized on the IMBs by the avidin–biotin conjugation. The assay was based on the change in the detection peak current. Under the optimum experimental conditions, the linear range of detection of CEA is of the two-component immunosensor is from 0.01 to 120 ng/mL, with a low detection limit of 0.003 ng/mL, and the linear range of detection of CA72-4 is from 0.05 to 35 U/mL, with a detection limit of 0.016 U/mL. The results showed that the proposed immunosensor enabled simultaneous monitoring of CEA and CA72-4 and exhibited good reproducibility, excellent high selectivity, and sensitivity. In particular, the proposed multiplexed immunoassay approach does not require sophisticated fabrication and is well-suited for high-throughput biosensing and application to other areas. Full article

Background: There is only limited research on the coronal cementum of a tooth, and the mechanisms of its forming process are not well-defined. This report presents a coronal cementum on the occlusal surfaces of enamel in an impacted wisdom tooth in a human, which is not nearly the cervical portion. Materials and Methods: The tooth (Tooth #1) was derived from a 46-year-old female. Histological analysis, including hematoxylin and eosin (HE) and toluidine blue (TB) staining, and Scanning Electron Microscopy and Energy Dispersive X-ray Spectrometer (SEM-EDS) analysis of the extracted tooth were conducted. Radiographic examination showed that Tooth #1 was horizontally impacted in the maxilla and had the apex of a single root placed between the buccal and palatal roots of Tooth #2. Results: Coronal cementum was distributed widely on the enamel, and reduced enamel epithelium was also found with enamel matrix proteins histologically. The formation of acellular cementum was observed to be more predominant than that of the cellular cementum in Tooth #1. SEM showed that the occlusal cementum connected directly with enamel. Calcium mapping revealed an almost similar occlusal cementum and enamel. In addition, the spectrum of elements in coronal cementum resembled the primary cementum according to SEM-EDS. Discussion: Thus, coronal cementogenesis in impacted human teeth might be related to the existence of reduced enamel epithelium. Full article

Significant increase in use of color dyes in modern society exerted a great pressure on environmental and water qualities. As such, studies for the removal of color dyes from water have been conducted extensively. In this study, common Earth materials dolomite and calcite were evaluated and contrasted for their removal of toluidine blue (TB), a cationic dye, before and after heat treatment. An increase by a factor of 3 in TB removal capacity from 3.5 to 10 mmol/kg was achieved after dolomite (Dol) was heated to 800 °C (designated as HDol). In contrast, the TB removal capacity increased by more than 100 times from 2 to 220 mmol/kg after calcite (Cal) was heated to 1000 °C (designated as HCal). For Dol and HDol, the TB removal increased as the solution’s pH increased but decreased with increases in the solution’s ionic strength. For Cal and HCal, the influence of the equilibrium solution’s pH and ionic strength on TB removal was negligible. The free energy of TB sorption on Dol, HDol, and Cal were −7 to −15 kJ/mol. The results suggested different removal mechanism for TB by Dol vs. Cal. X-ray diffraction data for Dol showed a slight increase in calcite content after heat treatment. For Cal, CaO was produced after heating, which converted back to calcite after 24 h of mixing with TB solutions. The significant TB removal by HCal could be attributed to its reaction with CaO. Thus, the best solution is to use freshly treated Cal for the removal of TB from solution. Full article

A wide variety of dyes, such as toluidine blue (TB), are used daily for a multitude of purposes. After use, many of these compounds end up in aqueous effluents, reaching natural environments, including marine environments. The removal of these pollutants from marine environments must be considered a priority problem. The search for natural techniques, such as biosorption, is a preferred option to eliminate pollution from natural environments. However, biosorption studies in seawater are scarce. For this reason, the living biomass of the marine microalga Phaeodactylum tricornutum was studied to determine its ability to remove TB from seawater. The kinetics of the biosorption process, the isotherms, and the effect of light and pH were determined. This biomass showed a maximum TB removal capacity of 45 ± 2 mg g⁻¹ in the presence of light. Light had a positive effect on the TB removal capacity of this living biomass. The best fitting kinetics was the pseudo-second order kinetics. The efficiency of the removal process increased with increasing pH. This removal was more effective at alkaline pH values. The results demonstrated the efficacy of P. tricornutum living biomass for the efficient removal of toluidine blue dye from seawater both in the presence and absence of light. Full article

Among the most malignant cancers, oral squamous cell carcinoma (OSCC) stands out as the most common malignant head and neck tumor. Despite advances in the field of treatment, the prognosis of patients with OSCC remains poor. Aiming to overcome the limitations of the currently existing therapies against OSCC, the present work aims to investigate the potential of photodynamic therapy (PDT) with phenothiazine derivatives used alone or in combination. The incorporation of methylene blue (MB) and toluidine blue (TB) was evaluated in OSCC cell lines (HSC-3 and SCC-9) and a nontumor cell line (Hfib). Both compounds exhibited concentration and time-dependent incorporation, with higher rates observed in tumor cells. Regarding dark-phase cytotoxic activity, SCC-9 cells were the most sensitive cell line with an IC₅₀ value of 362.6 µM and 41.4 µM for MB and TB, respectively. Using PDT, all lineages showed greater sensitivity, presenting lower IC₅₀ values when compared to the dark phase values. The combination index values of 0.69 (dark phase) and 0.73 (clear phase) associated with concave isobolograms, in both phases, revealed that MB and TB have synergistic effects when combined against SCC-9 cells. These findings suggest that MB or TB assisted with PDT holds promise for OSCC treatment. Full article

Although the fabrication of controllable three-dimensional (3D) microstructures on substrates has been proposed as an effective solution for SERS, there remains a gap in the detection and manufacturability of 3D substrates with high performance. In this study, photolithography is adopted to obtain a pyramid-like array on a patterned sapphire substrate (PSS), with Al₂O₃ as the dielectric layer. In addition, silver nanoparticles (AgNPs) are used to decorate Au films to obtain mass-producible 3D SRES substrates. In the case of low fluorescence, the substrate realizes the coupling of localized surface plasmon polaritons (LSPs) and surface plasmon polaritons (SPPs), which is consistent with the simulation results obtained using the finite element method. The performance of the SERS substrate is evaluated using rhodamine 6G (R6G) and toluidine blue (TB) as probe molecules with detection limits of 10⁻¹¹ M and 10⁻⁹ M, respectively. The substrate exhibits high hydrophobicity and excellent light-capturing capability. Moreover, it shows self-cleaning ability and long-term stability in practical applications. Allowing for the consistency of the composite substrate in the preparation process and the high reproducibility of the test results, it is considered to be promising for mass production. Full article

Herein, we designed chitosan–coated Fe₃O₄ nanocomposites for the control release of drugs by an alternating magnetic field (AMF). The chitosan-coated Fe₃O₄ nanoparticles (Fe₃O₄@CS) were prepared by a alkaline co-precipitation method, and then, the model drug toluidine blue (TB) was covalently grafted onto the surface of the nanocomposite by a two-step amide reaction with the thermosensitive molecule 4,4′-azobis (4-cyanovaleric acid) (ACVA) as the linker group. The prepared nanocomposites were superparamagnetic and showed high magnetization saturation (about 54.0 emu g⁻¹). In vitro hydrothermal release studies showed that most parts of the TB would be effectively enclosed within the nanocarriers at lower ambient temperatures (23 or 37 °C) due to the molecular bonding of ACVA. The results of kinetic fitting of hydrothermal release data showed that TB released from nanoparticles followed first-order kinetics (R² > 0.99) and the Korsemeyer–Peppas model (R² > 0.99, n < 0.5). Most importantly, a single magnetron release experiment demonstrated an approximately linear relationship between the cumulative release of the drug and the duration of action of AMF (R² = 0.9712). Moreover, the increase in the cumulative release of the drug can be controlled by controlling the switch of the AMF generation device. Therefore, the ACVA-modified Fe₃O₄@CS nanocarrier designed in this study is a promising model for drug delivery that enables the control of drug release dose by AMF. Full article

Lanthanum (La) nanocomposites LaFeO₃, LaNiO₃, and LaCoO₃ were synthesized using a sol-gel method, and different La to-metal (Fe, Ni, or Co) ratios were attained using various concentrations of salts. The resulting composites were calcined at 540 °C and characterized by XRD, SEM-EDX, FT-IR spectroscopy, XPS, thermogravimetric analysis (TGA), and PL spectroscopy. The activity of the lanthanum composites (LaFeO₃, LaNiO₃, and LaCoO₃) was studied using the photocatalytic degradation of methylene blue (MB) and ortho-toluidine blue (o-TB) under visible light with a wavelength below 420 nm. The change in the concentration of dyes was monitored by using the UV-Vis spectroscopy technique. All composites appeared to have some degree of photocatalytic activity, with composites possessing an orthorhombic crystal structure having higher photocatalytic activity. The LaCoO₃ composite is more efficient compared with LaFeO₃ and LaNiO₃ for both dyes. High degradation percentages were observed for the La composites with a 1:1 metal ratio. Full article

Snail shells (Anadora Fulica) calcined at different temperatures were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, thermal analyses (TG-DTG), scanning electron microscopy (SEM) and N₂ adsorption–desorption experiments (surface area measurements were found using the coupled BET/BJH method). The principal objective was to identify different forms of calcium carbonate and calcium hydroxide in snail shells as raw materials. The calcium hydroxide thus obtained was used in the synthesis of the hydroxyapatite/L-lysine (HA/Lys) composite. The composite used to chemically modify a glassy carbon electrode (GCE) was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). It appeared that the developed sensor Lys/HA/GCE facilitated electronic transfer compared to the pristine electrode. In a strongly acid medium, this surface protonated and therefore became positively charged, which allowed it to have a good affinity with [Fe(CN)₆]^3-. An application in toluidine blue (TB) electroanalysis in the phosphate buffer was carried out. Optimal sensor performances were obtained using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The performance of the sensor was determined in the concentration range 1 to 10 µM of TB, and the limit of detection (LOD) obtained by the S/N = 3 method was 2.78 × 10⁻⁷ M. The sensor was also used to detect the TB in spring water at 96.79% recovery. Full article

In this study, we prepared spheroid microstructures of monoclinic bismuth phosphate BiPO₄ by a facile solid-state reaction at 500 °C. The crystal structure was refined using the Rietveld method, where the crystal cell was resolved using a monoclinic system (parameters a, b, c, β) with space group P2₁/n. SEM images showed that the solid catalyst presented homogeneous morphologies. These BiPO₄ microparticles (BiP-500) have been used as photocatalysts to photodegrade, under UV light irradiation, three cationic dyes (Rhodamine B, RhB; Methylene Blue, MB; and Toluidine Blue, TB), three anionic dyes (Congo Red, CR; Orange G, OG; and Methyl Orange, MO) and mixtures of RhB-MB, RhB-OG and MO-OG organic dyes. The photodegradation efficiency of these BiP-500 microparticles is found to be optimal in the case of RhB solutions, RhB-MB and RhB-OG binary mixtures. The BiP-500 catalyst shows a high selectivity for the conversion of the mixture of dyes into CO₂ and H₂O. Total organic carbon analysis of an anionic dye and a cationic dye (RhB, OG and RhB-MB) confirms the mineralization of the pollutants in the presence of BiP-500 particles. The photocatalytic efficiency of our BiP-500 photocatalyst has been confirmed, with a view to facilitate applications in the field of the depollution of wastewater in the agricultural environment by the degradation of parathion-methyl (PM) as a pollutant. Full article

Mast cells (MCs) have relevant participation in inflammatory and vascular hyperpermeability events, responsible for the action of the kallikrein–kinin system (KKS), that affect patients inflicted by the severe form of COVID-19. Given a higher number of activated MCs present in COVID-19 patients and their association with vascular hyperpermeability events, we investigated the factors that lead to the activation and degranulation of these cells and their harmful effects on the alveolar septum environment provided by the action of its mediators. Therefore, the pyroptotic processes throughout caspase-1 (CASP-1) and alarmin interleukin-33 (IL-33) secretion were investigated, along with the immunoexpression of angiotensin-converting enzyme 2 (ACE2), bradykinin receptor B1 (B1R) and bradykinin receptor B2 (B2R) on post-mortem lung samples from 24 patients affected by COVID-19. The results were compared to 10 patients affected by H1N1pdm09 and 11 control patients. As a result of the inflammatory processes induced by SARS-CoV-2, the activation by immunoglobulin E (IgE) and degranulation of tryptase, as well as Toluidine Blue metachromatic (TB)-stained MCs of the interstitial and perivascular regions of the same groups were also counted. An increased immunoexpression of the tissue biomarkers CASP-1, IL-33, ACE2, B1R and B2R was observed in the alveolar septum of the COVID-19 patients, associated with a higher density of IgE⁺ MCs, tryptase⁺ MCs and TB-stained MCs, in addition to the presence of intra-alveolar edema. These findings suggest the direct correlation of MCs with vascular hyperpermeability, edema and diffuse alveolar damage (DAD) events that affect patients with a severe form of this disease. The role of KKS activation in events involving the exacerbated increase in vascular permeability and its direct link with the conditions that precede intra-alveolar edema, and the consequent DAD, is evidenced. Therapy with drugs that inhibit the activation/degranulation of MCs can prevent the worsening of the prognosis and provide a better outcome for the patient. Full article

Bacterial infections in wounded skin are associated with high mortality. The emergence of drug-resistant bacteria in wounded skin has been a challenge. Toluidine blue (TB) is a safe and inexpensive photosensitizer that can be activated and used in near-infrared photodynamic therapy to effectively kill methicillin-resistant Staphylococcus aureus (MRSA). However, its aggregation-induced quenching effect largely affects its clinical applications. In this study, TB nanoparticles (NPs) were synthesized using an ultrasound-assisted coating method. Their physicochemical and biological properties were studied and evaluated by scanning electron microscopy and Fourier-transform infrared spectroscopy. The TBNPs had a broad-spectrum antibacterial activity against Gram-positive bacteria (MRSA) and Gram-negative bacteria (E. coli). In addition, MTT, hemolysis, and acute toxicity tests confirmed that TBNPs had good biocompatibility. The TBNPs exhibited a high photodynamic performance under laser irradiation and efficiently killed E. coli and MRSA through generated reactive oxygen species, which destroyed the cell wall structure. The potential application of TBNPs in vivo was studied using an MRSA-infected wound model. The TBNPs could promote wound healing within 7 days, mainly by reducing the inflammation and promoting collagen deposition and granulation tissue formation. In conclusion, the TBNPs offer a promising strategy for clinical applications against multiple-drug resistance. Full article