Cellulose extraction of Alstonia scholaris: A comparative study on efficiency of different bleaching reagents for its isolation and characterization.

The incredible benefits of Alstonia scholaris are piquing researchers' attention in extracting its cellulose and utilizing it in further therapeutic applications. This study is based on cellulose extraction from its stalks and processed through chemical pre-treatments to manifest its cellulose content by using different bleaching reagents. A comparison was made on efficiencies of three reagents and it is found that the hydrogen peroxide exposed maximum cellulose than sodium hypochlorite and sodium chlorite. The experimental results revealed that A. scholaris possess 68-70% cellulose content. FTIR spectrum shows that OH- and CH- vibrations of cellulose appeared at 3320 cm-1 & 2892 cm-1 respectively whereas SEM images show fibrillation, rough surface, and lumens in bleached fiber that attributes to the removal of lignin and hemicelluloses and confirms cellulose extraction. The XRD pattern certifies the crystalline nature and compactness of cellulose whereas tensile properties and TGA help in understanding its flexibility, mechanical strength, and thermal stability at 370 °C respectively.

Evaluating the Antioxidants, Whitening and Antiaging Properties of Rice Protein Hydrolysates.

Plant-derived protein hydrolysates have potential applications in nutrition. Rice protein hydrolysates (RPHs), an excellent source of proteins, have attracted attention for the development of cosmeceuticals. However, few studies have reported the potential application of RPH in analysis, and this study examined their antioxidant activities and the inhibitory activities of skin aging enzymes. The results indicated that the total phenolic and flavonoid concentrations were 2.06 ± 0.13 mg gallic acid equivalent/g RPHs and 25.96 ± 0.52 µg quercetin equivalent/g RPHs, respectively. RPHs demonstrated dose-dependent activity for scavenging free radicals from 1,1-diphenyl-2-picrylhydrazyl [half-maximal inhibitory concentration (IC50) = 42.58 ± 2.1 mg/g RPHs] and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (IC50 = 2.11 ± 0.88 mg/g RPHs), dose-dependent reduction capacity (6.95 ± 1.40 mg vitamin C equivalent/g RPHs) and oxygen radical absorbance capacity (473 µmol Trolox equivalent/g RPHs). The concentrations of the RPH solution required to achieve 50% inhibition of hyaluronidase and tyrosinase activities were determined to be 8.91 and 107.6 mg/mL, respectively. This study demonstrated that RPHs have antioxidant, antihyaluronidase, and antityrosinase activities for future cosmetic applications.

Partial exchange of ozone by electron beam irradiation shows better viscosity control and less oxidation in cellulose upgrade scenarios.

Electron beam irradiation (EBI) is an alternative treatment for intrinsic viscosity (IV) control in cellulose pulps, but has never been integrated in full bleaching sequences for comparison to conventional methods. Both eucalyptus kraft (EK) paper pulp and beech sulfite (BS) dissolving pulp were subjected to totally chlorine free (TCF) bleaching sequences comprising either EBI, ozone, or both for IV control. Additionally, effects of EBI on hexenuronic acid (HexA) and xylan were investigated. IV was adjusted to 450-500 mL g-1 and properties including carbonyl content, kappa, brightness, alkali-resistance and sugar composition were compared. Pulps produced with EBI had a higher alkali-resistance, uniformity and less cellulose oxidation. However, the degree of bleaching (DoB) was low without the use of ozone. HexA content in a birch pulp was halved by EBI. Isolated xylans were more resistant to irradiation than cellulose with little decrease of molar masses and moderate oxidation.

All-biobased transparent-wood: A new approach and its environmental-friendly packaging application.

Transparent-wood (TW) is an emerging research topic that can be applied to biobased products. However, it is necessary to fill pores in TW with natural substances to prepare all-biobased TW. This paper reports an all-biobased TW by infiltrating cellulose nanofiber (CNF) and chitosan (CTS) suspensions into the bleached wood. CNF was isolated by combining the chemical and physical methods, and CTS was dissolved in acetic acid, and they were infiltrated into the pores of the bleached Fir veneer wood using a vacuum jar. The CNF and chitosan effects on the mechanical properties of the TW were studied, and the morphologies, crystallinity index, water contact angle, antioxidant, thermal degradation, and UV-shielding properties were investigated. The prepared TW showed 80 % total transmittance and 30-60 % haze, suitable for solar cell application. The all-biobased TW showed good thermal stability up to 315 °C and excellent UV shielding property for UV-B and UV-C. The antioxidant property of the CTS-TW significantly increased as compared to the original wood. The CNF-TW showed considerable tensile strength and yield strength of more than 200 % improved from the original wood. The potential for environment-friendly packaging applications was demonstrated by making a bag, medicine packaging, and straw for a drink.

Immobilization of peroxidase on textile carrier materials and their application in the bleaching of colored whey.

Textiles represent promising support materials for enzymes. The goal of the present work was to investigate the immobilization of commercial peroxidase on a polyester needle felt and the repeated use in the gentle degradation of norbixin in whey from dairy cheese as a practical application. High enzyme loads were obtained by a 2-step immobilization procedure. First, the number of functional groups on the textile surface was increased by a modification with amino-functional polyvinylamine. Second, the enzyme was immobilized by using 2 types of crosslinking agents. Due to the iron content of peroxidase, inductively coupled plasma-optical emission spectrometry was used for the quantitative determination of the enzyme load on the textile. The enzyme activity was evaluated using common 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) assay for peroxidases. By the variation of enzyme input and crosslinker concentration, a maximal enzyme load of 80 mg/g of textile was achieved, and a maximum specific activity of 57 U/g of textile. For the visualization of the enzyme on the fiber surface, fluorescence microscopy as well as scanning probe microscopy were used. The immobilized peroxidase showed significant activity, even after 50 reuse cycles. In addition, the potential of the new support and enzyme combination in commercial whey bleaching was demonstrated successfully on a 10-L scale.

Optimisation of the Production and Bleaching Process for a New Laccase from Madurella mycetomatis, Expressed in Pichia pastoris: from Secretion to Yielding Prominent.

Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these "green catalysis" enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were heterologously expressed in the methylotrophic yeast host Pichia pastoris. The high yield of the active recombinant protein in P. pastoris demonstrates the efficiency of a reliably constructed plasmid to express the laccase gene. The optimal biochemical conditions for the successfully expressed MmLac enzyme were identified. Detailed structural properties of the recombinant laccase were determined, and its utility in decolourisation and textile bleaching applications was examined. MmLac demonstrates good activity in an acidic pH range (4.0-6.0); is stable in the presence of cationic metals, organic solvents and under high temperatures (50-60 °C); and is stable for long-term storage at - 20 °C and - 80 °C for up to eight weeks. The structural analysis revealed that the catalytic residues are partially similar to other laccases. MmLac resulted in an increase in whiteness, whilst demonstrating high efficiency and stability and requiring the input of fewer chemicals. The performance of this enzyme makes it worthy of investigation for use in textile biotechnology applications, as well as within environmental and food technologies.

A DyP-Type Peroxidase of Pleurotus sapidus with Alkene Cleaving Activity.

Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete Pleurotus sapidus (PsaPOX) cleaved the aryl alkene trans-anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn2+-oxidation site on its surface. The gene was expressed in Komagataella pfaffii GS115 yielding activities up to 142 U/L using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H2O2 and 25 mM Mn2+. PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn2+ to Mn3+. In addition, bleaching of ß-carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes (E)-methyl isoeugenol, α-methylstyrene, and trans-anethole, which was increased almost twofold in the presence of Mn2+. The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.

Effects of 16% Carbamide Peroxide Bleaching on the Surface Properties of Glazed Glassy Matrix Ceramics.

OBJECTIVE: To determine the influence of the home bleaching agent, Opalescence PF, on the surface roughness and microhardness of glazed glassy matrix CAD-CAM ceramics. Materials and Methods. The 28 sintered leucite- and lithium disilicate-reinforced ceramic specimens (IPS Empress CAD and IPS e.max CAD) were divided into control and bleached groups. The home bleaching agent was applied to specimens of bleached groups for 7 days. The surface roughness and microhardness of all specimens were measured. A scanning electron microscope was used to evaluate the surface properties. The data were statistically analyzed by two-way ANOVA. RESULTS: The control e.max CAD showed the lowest surface roughness values. For both Empress and e.max CAD, surface roughness was significantly higher for the bleached group (p < 0.05). No significant differences in microhardness were observed. CONCLUSIONS: According to our study, patients should be careful when using home bleaching agents because whitening agents can affect the mechanical properties of full ceramic restorations like e.max CAD and Empress CAD. Ceramic polishing may be required in clinical situations where ceramic restorations are accidentally exposed to bleaching gels.

The effect of bleaching agents on the DNA analysis of bloodstains on different floor coverings.

Blood at crime scenes is one of the most significant traces of evidence in investigation proceedings. Cleaning up these traces with household cleaning products, often containing bleaching agents, inhibits or complicates the detection of DNA. In this study, human blood was applied onto different floor coverings (carpet, laminate, parquet, PVC, tile) and subsequently cleaned with water and bleaching agents (hydrogen peroxide, sodium hypochlorite, DanKlorix®, Vanish Oxi Action®) at different times. Samples have been collected afterwards from the floors. The samples underwent a quantitative and qualitative DNA analysis. Cleaning smooth surfaces with water is usually sufficed to prohibit retrieving a DNA profile in most of the cases. Cleaning carpets was more difficult due to their absorbent surface whereas the use of bleaching agents caused an additional reduction of verifiable DNA concentrations. Retrieving partial or complete profiles after the use of bleaching agents was only possible when cleaning with low concentrations of 3% hydrogen peroxide.

Xylanases from marine microorganisms: A brief overview on scope, sources, features and potential applications.

Global economic growth often leads to depletion of raw materials and generation of greenhouse gases, as industry manufactures goods at ever increasing levels to keep up with the demand. The currently implemented production processes mostly rely on non-renewable resources, they suffer from high energy consumption, and generate waste that often has a negative environmental impact. Eco-friendly production methods are therefore intensely searched for. Among them, enzyme-based processes are appealing, because of their high substrate and reaction specificity and the relatively mild operation conditions required by these catalysts. In addition, renewable raw materials that allow sustainable production processes are also widely explored. Marine xylanases, which catalyze the hydrolysis of xylan, the major component of lignocellulose, are promising biocatalysts. Since they are produced by microorganisms that thrive in a wide variety of environmental conditions, the enzymes may be active at widely different ranges of pH, temperature, and salt concentrations. These properties are important for their successful application in various industrial processes, such as production of bioethanol, bleaching of paper and pulp, and in the food and feed sector. The present work gives a brief overview of marine sources of xylanases, their classification and features, and of the potential applications of these marine enzymes, especially in sustainable processes in the scope of circular economy.

Size-dependent whitening activity of enzyme-degraded fucoidan from Laminaria japonica.

Fucoidan from Laminaria japonica is a kind of sulfate polysaccharide with high molecular weight (MW) and broad bioactivities. This study was performed to investigate the relationship between MW and whitening activity of fucoidan and to exploit a novel functional ingredient for whitening cosmetics. High sulfate content fucoidan was enzymic degraded by Flavobacterium RC2-3 produced fucoidanase. Two hours were enough for the enzyme degradation to achieve degraded fucoidan with favorable tyrosinase inhibitory ability. The whitening activity of different MW fucoidan fractions were evaluated by their tyrosinase inhibitory ability, antioxidant activity and cellular melanogenesis inhibitory ability. Results showed that in the MW range above 5 kDa, the smaller MW of fucoidan were related to the better whitening activity. The fucoidan fraction with the MW between 5-10 kDa, presented the best tyrosinase inhibitory activity (62.0%), antioxidant activity (48.3%) and excellent anti-melanogenesis ability in B16 cells, which could be applied as the whitening factor in cosmetics development.

Effect of Polymerization Time and Home Bleaching Agent on the Microhardness and Surface Roughness of Bulk-Fill Composites: A Scanning Electron Microscopy Study.

OBJECTIVE: The aim of this study is to evaluate the microhardness and surface roughness of two different bulk-fill composites polymerized with light-curing unit (LCU) with different polymerization times before and after the application of a home bleaching agent. MATERIALS-METHODS: For both microhardness and surface roughness tests, 6 groups were prepared with bulk-fill materials (SonicFill, Filtek Bulk Fill) according to different polymerization times (10, 20, and 30 s). 102 specimens were prepared using Teflon molds (4 mm depth and 5 mm diameter) and polymerized with LCU. 30 specimens (n = 5) were assessed for microhardness. Before home bleaching agent application, the bottom/top (B/T) microhardness ratio was evaluated. After bleaching agent application, the microhardness measurements were performed on top surfaces. Roughness measurements were performed in 72 specimens (n = 12) before and after bleaching application. Additionally, for SEM analyses, two specimens from all tested groups were prepared before and after bleaching agent application. The data B/T microhardness ratio before bleaching was analyzed by two-way ANOVA and Tukey's HSD test. The data from the top surface of specimens' microhardness before and after bleaching were analyzed using Wilcoxon signed-rank test, Kruskal-Wallis, Mann-Whitney U tests. The data from surface roughness tests were statistically analyzed by multivariate analysis of variance and Bonferroni test (p < 0.05). RESULTS: The B/T microhardness ratio results revealed no significant differences between groups (p > 0.05). Comparing the microhardness values of the composites' top surfaces before and after bleaching, a significant decrease was observed exclusively in FB30s (p < 0.05). No significant differences in surface roughness values were observed when the groups were compared based on bulk-fill materials (p > 0.05) while the polymerization time affected the surface roughness of the SF20s and SF30s groups (p < 0.05). After bleaching, surface roughness values were significantly increased in the SF20s and SF30s groups (p < 0.05). CONCLUSION: The clinicians should adhere to the polymerization time recommended by the manufacturer to ensure the durability of the composite material in the oral environment.

Quantification of carbonate radical formation by the bicarbonate-dependent peroxidase activity of superoxide dismutase 1 using pyrogallol red bleaching.

Carbonate radicals (CO3-) are generated by the bicarbonate-dependent peroxidase activity of cytosolic superoxide dismutase (Cu,Zn-SOD, SOD-1). The present work explored the use of bleaching of pyrogallol red (PGR) dye to quantify the rate of CO3- formation from bovine and human SOD-1 (bSOD-1 and hSOD-1, respectively). This approach was compared to previously reported methods using electron paramagnetic resonance spin trapping with DMPO, and the oxidation of ABTS (2,2-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid). The kinetics of PGR consumption elicited by CO3- was followed by visible spectrophotometry. Solutions containing PGR (5-200 µM), SOD-1 (0.3-3 µM), H2O2 (2 mM) in bicarbonate buffer (200 mM, pH 7.4) showed a rapid loss of the PGR absorption band centered at 540 nm. The initial consumption rate (Ri) gave values independent of the initial PGR concentration allowing an estimate to be made of the rate of CO3- release of 24.6 ±â€¯4.3 µM min-1 for 3 µM bSOD-1. Both bSOD-1 and hSOD-1 showed a similar peroxidase activity, with enzymatic inactivation occurring over a period of 20 min. The single Trp residue (Trp32) present in hSOD-1 was rapidly consumed (initial consumption rate 1.2 ±â€¯0.1 µM min-1) with this occurring more rapidly than hSOD-1 inactivation, suggesting that these processes are not directly related. Added free Trp was rapidly oxidized in competition with PGR. These data indicate that PGR reacts rapidly and efficiently with CO3- resulting from the peroxidase activity of SOD-1, and that PGR-bleaching is a simple, fast and cheap method to quantify CO3- release from bSOD-1 and hSOD-1 peroxidase activity.

Semi-automated standardisation of melanin bleaching procedures of heavily pigmented melanocytic lesions for immunohistochemical analysis on an automated platform.

Background: The diagnosis of heavily pigmented melanocytic lesions is problematic. This is often compounded by lack of visibility of nuclear detail of tumour cells due to physical masking by melanin pigment. Similarly, there can be colour merging of chromogenic final reaction products with melanin, making an evidence of antigenic localisation problematic. There are a number of melanin bleaching techniques available for immunohistochemical assessments.Material and methods: All methods to date have involved the bleaching of melanin as a manually performed primary step before loading subsequently bleached slides onto automated immunohistochemical platforms. Here we define a semi-automated bleaching procedure that allows full integration on one of the most widely employed automated IHC staining platforms (Roche Ventana BenchMark Ultra). The bleaching protocol was defined on the BenchMark Ultra and involved the assessment of 24 histological cases of heavily pigmented malignant melanoma lesions (13 cutaneous and 11 metastatic) routinely fixed processed and paraffin wax embedded.Results: Completion of the bleaching was assessed on H&E preparations performed following the semi-automated bleaching step and employing the Roche Ventana BenchMark Ultra machine for 60 min at 42°C. Complete immunohistochemical staining was achieved on the automated platform within 5-6 h including the bleaching step. Results were consistent across all tissue evaluated.Discussion: This data provides evidence that the hydrogen peroxide bleaching procedure can be adapted for integration on one of the most widely employed automated IHC staining platforms and as a result, improve the efficiency and reproducibility of the technique.

Deionized water can substitute common bleaching agents for nonvital tooth bleaching when treated with non-thermal atmospheric plasma.

The bleaching efficacy of common bleaching agents and deionized water treated with non-thermal atmospheric pressure plasma in the pulp chamber for nonvital tooth bleaching was evaluated. A total of 120 extracted human maxillary first incisors were stained using human blood. Teeth were randomly divided into eight groups (n = 15). In the first four groups, teeth were bleached using 35% hydrogen peroxide gel, 37% carbamide peroxide gel, 2:1 (w/v) sodium perborate paste, and deionized water for 30 min. In the remaining groups, bleaching agents were treated with non-thermal atmospheric plasma for 5 min inside the pulp chamber. Overall color changes (∆E) were determined using Commission Internationale de L'Eclairage Lab Colour System. The plasma-assisted tooth bleaching has not increased tooth temperature beyond 37°C. Bleaching efficacies of bleaching agents were significantly improved when treated with non-thermal atmospheric plasma compared to their application (P < 0.05). A remarkable bleaching effect was obtained when bleaching agents were substituted with water and when treated with non-thermal atmospheric plasma. Non-thermal atmospheric plasma treatment could be a novel tool for activation of bleaching agents in the pulp chamber for nonvital tooth bleaching procedure. Moreover, water could be used as a novel bleaching agent when treated with the non-thermal atmospheric plasma to eliminate possible risks which might arise from peroxide-containing agents.

L-765,314 Suppresses Melanin Synthesis by Regulating Tyrosinase Activity.

Although melanin production is a key self-defense mechanism against ultraviolet radiation (UVR)-induced skin damage, uneven or excessive deposition of melanin causes hyperpigmentary disorders. Currently available whitening agents are unsatisfactory because of issues with efficacy and safety. To develop more effective depigmenting agents, we performed high-throughput melanin content assay screening using the B16F10 melanoma cell line and identified L-765,314 as a drug that suppressed melanin production in cultured melanocytes in a dose-dependent manner as well as cAMP- or 12-O-tetradecanoylphorbol 13-acetate (TPA)-stimulated melanin production without cytotoxicity. Interestingly, melanogenic gene expression was not altered by L-765,314. Rather, diminished melanin production by L-765,314 appeared to be caused by downregulation of tyrosinase activity via inhibition of protein kinase C (PKC). Because L-765,314 did not show any adverse effect in melanocytes, altogether our data suggest that L-765,314 could be a potential therapeutic candidate for skin hyperpigmentary disorders and further discovery of selective inhibitors targeting PKC might be a promising strategy for the development of depigmenting agents to treat hyperpigmentary disorders.

Novel colour additive for bleach disinfectant wipes reduces corrosive damage on stainless steel.

Bleach disinfectant wipes are corrosive to hospital surfaces and equipment. This study measured the effect of two widely used bleach wipes, with and without Highlight® colour additive, on stainless steel to quantify the rate of corrosion and to determine the effect of Highlight® on reducing surface damage caused by bleach wipes. The two bleach wipes alone caused severe corrosion [>5 mils per year (mpy), where 1 mil = 0.001 inch], while the addition of Highlight® reduced the rate of corrosion significantly (<2 mpy) and prevented discolouration of the metal. These results indicate that Highlight® reduces the deleterious corrosive effects of bleach wipes, thus improving their viability for cleaning healthcare surfaces.

The presence of fecal test soil protects bacteria during cleaning/disinfection.

Reusable medical devices (RMDs) must be reprocessed between uses to render them safe for each use and each patient. Cleaning used devices removes organic and inorganic soil making them either safe for reuse or ready for disinfection/sterilization depending on the device. Although cleaning is an important step in a RMD's life cycle, it is not always a priority during device design. In addition, when performing cleaning validation, it is recommended that the manufacturer takes into consideration, what the most appropriate or worst case conditions are in terms of type of soil or the presence of bacteria. This study compared the ability of three different cleaning/disinfecting agents (water, alcohol, and bleach) to remove bacteria and fecal test soil from two different polymers: polypropylene and ultrahigh molecular weight polyethylene (UHMWPE) with two different roughness. There were some differences in the effects of the cleaning/disinfecting agents, the materials, and the roughness depending on the particular circumstances. However, the most consistent effect on the removal of bacteria was the presence of soil, which protected the bacteria from being removed. Conversely, the presence of bacteria played little role in the removal of soil. Although the interactions between material type and roughness, soil type, and bacteria are complicated, they should be taken into account during device design and reprocessing validation to create a device that is easy and safe to use. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1706-1710, 2019.

Effects of household bleach on sputum smear microscopy to concentrate acid fast bacilli for the diagnosis of pulmonary tuberculosis.

Tuberculosis (TB) is one of the major public health problem among contagious diseases in Pakistan. TB diagnosis mainly depends on sputum smear microscopy. The main objective of this study was to evaluate the effects of household bleach on sputum smear microscopy to concentrate acid fast bacilli for the diagnosis of pulmonary tuberculosis. Sputum specimens of 200 suspected TB patients were collected for the study. Smears were prepared from the purulent part of sputum sample before and after bleach treatment, heat fixed and stained with the ZN technique. The obtained data were analyzed by chi-squared test using SPSS software. Out of 200 isolates, 22 (11%) patients had positive smears for acid fast bacilli (AFB) by direct ZN staining. After treatment with household bleach (NaOCL) and centrifugation, the number of AFB positive patients were increased from 22 (11%) to 37 (18.5%). The bleach-concentration method for sputum samples significantly increased the TB detection rate as compared to direct sputum smear microscopy. Thus, a shift from direct sputum microscopy to bleach-concentration technique should be considered a better method for detection of AFB in sputum through smear microscopy.

Melanin Bleaching With Warm Hydrogen Peroxide and Integrated Immunohistochemical Analysis: An Automated Platform.

OBJECTIVE: Diagnosing melanocytic lesions is among the most challenging problems in the practice of pathology. The difficulty of physically masking melanin pigment and the similarity of its color to commonly used chromogens often complicate examination of the cytomorphology and immunohistochemical staining results for tumor cells. Melanin bleach can be very helpful for histopathological diagnosis of heavily pigmented melanocytic lesions. Although various depigmentation methods have been reported, no standardized methods have been developed. This study developed a fully automated platform that incorporates hydrogen peroxide-based melanin depigmentation in an automated immunohistochemical analysis. METHODS AND MATERIALS: The utility of the method was tested in 1 cell block of malignant melanoma cells in pleural effusion, 10 ocular melanoma tissue samples, and 10 cutaneous melanoma tissue samples. Our results demonstrated that the proposed method, which can be performed in only 3 hours, effectively preserves cell cytomorphology and immunoreactivity. RESULTS: The method is particularly effective for removing melanin pigment to facilitate histopathological examination of cytomorphology and for obtaining an unmasked tissue section for immunohistochemical analysis.