Plant melanin, a natural pigment, has gained significant attention recently due to its potential therapeutic and industrial applications. In this study, melanin pigments were extracted from Nigerseed hulls (NH) via alkali and acid extraction methods, followed by acid hydrolysis, organic solvent treatment, and repeated precipitation. The solubility of NH melanin was assessed, revealing solubility in alkali and dimethyl sulfoxide (DMSO) but insolubility in other common organic solvents. High-performance liquid chromatography (HPLC) was employed to measure the purity of NH melanin in comparison to standard melanin, while elemental analysis indicated a similarity between melanin extracted from nigerseed hulls and the standard counterpart. LC-MS data revealed a molecular weight of NH melanin. Furthermore, the stability of melanin was evaluated under varying conditions including temperature, oxidants, reducing agents, light exposure, and metal ion presence. Results demonstrated significant effects of Mg2+, Cu2+, and Fe2+ metal ions on melanin stability, with a minor effect observed for Ca2+, while sodium hyposulfite was found to destabilize the pigments. Our findings suggest that nigerseed hulls hold promise as a novel source for efficient melanin production, with potential applications in the food sector, food packaging, and biomedical fields.

Owing to the growing need for natural materials in different fields, studying melanin production from biological sources is imperative. In the current study, the extracellular melanin pigment was produced by the fungus Curvularia soli AS21 ON076460. The factors that affect the production of melanin were optimized by the Plackett-Burman design (P-BD). The effect of gamma irradiation on melanin productivity was investigated. The maximum melanin yield (3.376 mg/L) was elicited by a stimulus of gamma irradiation at 1.0 kGy. The results evoked that, Curvularia soli AS21 ON076460 melanin exhibited excellent antimicrobial activity against all tested bacteria and fungi. Klebsiella pneumoniae ATCC 13883 and P. digitatum were mostly affected by melanin registering the inhibition zone diameters of 37.51 ± 0.012 and 44.25 ± 0.214 mm, respectively. Moreover, Curvularia soli AS21 ON076460 melanin indicated a significant antiviral efficacy (77% inhibition) of Herpes simplex virus (HSV1). The melanin pigment showed antioxidant activities with IC50 of 42 ± 0.021 and 17 ± 0.02 µg/mL against DPPH and NO, respectively. Melanin had cytotoxic action against human breast cancer and skin cancer cell lines (Mcf7and A431) as well as exerting a low percentage of cell death against normal skin cell lines (Hfb4). Melanin was effective in wound management of human skin cells by 63.04 ± 1.83% compared with control (68.67 ± 1.10%). The novelty in the study is attributed to the possibility of using gamma rays as a safe method in small economic doses to stimulate melanin production from the fungi that have been isolated. In summary, melanin produced from fungi has significant biological activities that encourage its usage as a supportive medical route.

Introduction. The difficulty in diagnosis of severe melanocytic lesions is a problem to be overcome in pathological practice. Melanin bleaching is an effective approach to ameliorate melanin disturbances in severely pigmented lesions. Although various methods for improving melanin pigmentation in immunohistochemical staining have been reported, these depigmentation methods still need to be optimized and standardized. In this study, the coloring efficiency of 3,3\'-diaminobenzidine (DAB) and alkaline phosphatase (AP) after melanin depigmentation was compared under the automatic immunohistochemical staining platform. Methods. The applicability of the optimized depigmentation method was validated in 10 formalin-fixed paraffin-embedded (FFPE) blocks of ocular melanoma tissues. Specimens were demelaninized with 10% hydrogen peroxide at 60°C for immunohistochemical staining (Melan-A and SOX10), and tissue chromogenic staining was performed with DAB and AP detection systems, respectively. Results. The optimized depigmentation method including immunohistochemistry (IHC) could be completed in 3 h, effectively preserving cell morphology and immunoreactivity. Among these, the color-rendering effect and contrast of AP are better than DAB. Conclusion. This optimized method can effectively remove melanin and improve the accuracy of IHC staining interpretation. AP staining has better visibility and readability without the interference of residual melanin. The comparison results showed that after melanin depigmentation, the immunohistochemical staining agent was replaced with red AP, which avoided the misjudgment caused by brown DAB when melanin depigmentation was incomplete. This improved method can be applied to future histopathological and immunohistochemical staining of melanin-deposited tissues.

In the central nervous system, the presence of pigment in astrocytic tumors is rare. Only nine cases were reported in the English literature: one ganglioglioma, one pilocytic astrocytoma, and seven cases of pleomorphic xanthoastrocytoma (PXA). Though squash cytology is a common and useful tool for intraoperative diagnosis during brain tumor surgeries, the cytologic findings of pigmented PXA have not been recorded previously. We present a 32-year-old woman with a mass in her medial right temporal lobe. Intraoperative squash cytology examination demonstrated pleomorphic tumor cells containing cytoplasmic pigment granules. The subsequent tissue section and additional workup revealed a PXA with melanosomal melanin pigment deposits. While conventional PXA has to be differentiated from high-grade tumors such as glioblastoma, in the pigmented variant the presence of pigment can cause further diagnostic confusion with other pigmented tumors, in particular melanoma. It should be added into the differential diagnoses when a brain tumor smear shows nuclear pleomorphism and intracytoplasmic pigment particles.

Building the biological profile of a deceased person represents a pivotal step in order to achieve the victim\'s identification. Specifically with regard to ancestry, the melanin distribution pattern in the dermal-epidermal layers has been poorly explored in the forensic field as a potential useful tool. In particular, nothing has been reported about the reliability of such method in bodies in active decay or in advanced state of decomposition. In this study fragments of skin sampled from bodies of known ancestry, both in good and in poor states of preservation, were subjected to histological analysis. We selected 15 subjects, which were divided into three groups: group A (5 white Europeans), group B (5 black Africans) and group C (5 Orientals). A double skin sample was performed on all the bodies, one from the abdomen and the other one from the right forearm. After histological processing and staining with hematoxylin and eosin (H&E) and Masson-Hamperl trichrome technique, the cutaneous melanin distribution pattern was assessed using a semi-quantitative score. The melanin distribution patterns observed both in fresh and in putrefied cadaveric skin were found to be in all cases consistent with the victims\' known ancestry. Moreover no differences were observed between abdominal and forearm skin samples and all the histological findings highlighted by H&E were confirmed by the Masson-Hamperl trichrome staining. We demonstrated that the histological analysis aimed at assessing the melanin distribution pattern may be a valuable useful tool in the assessment of ancestry.

Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg2+ ions and the insoluble PPM is dominated by Ca2+ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.

Only one case of melanoma arising from melanin-producing medullary thyroid carcinoma (MTC) has been reported previously. In the present study, a second such case was reported and compared with the previous one. The patient was an 86-year-old male who presented with a right anterior neck mass. Ultrasound revealed a nodule measuring 49x48x40 mm in the right lobe of the thyroid. The levels of serum calcitonin (2,298 pg/ml) and carcinoembryonic antigen (CEA; 27.0 ng/ml) were markedly elevated. Aspiration cytology revealed suspected malignant anaplastic thyroid carcinoma and total thyroidectomy without neck nodal dissection was performed. On gross observation, the nodule was well encapsulated, soft, solid and black. Light microscopy indicated that the nodule was composed mainly of large, occasionally huge, pleomorphic cells with a solid or alveolar growth pattern. On immunohistochemistry, these cells were positive for melan-A and S-100 protein, and negative for thyroid transcription factor 1, calcitonin, chromogranin A and CEA. In the subcapsular area, melanin-producing MTC was intimately intermingled with the pleomorphic cells. No primary site of the melanoma was detectable in other organs. At three years after surgery, the patient died due to metastasis of the melanoma to the brain. The previously reported case had no detectable recurrence or distant metastasis up to 11 years after surgery. In comparison with that case, the present case had a similar morphology but the outcome was poorer. Thus, the prognosis of melanoma that transforms from MTC appears to remain uncertain.

Eosinophilic solid and cystic renal cell carcinoma (ESC-RCC) is an emerging entity in renal neoplasia with distinctive histopathological findings and a generally favorable prognosis. The presence of melanin pigment in a renal tumor typically prompts the observer to consider the microphthalmia-associated transcription family translocation renal cell carcinomas. We present a renal tumor occurring in a 19-year-old male patient which had the typical morphology of ESC-RCC but showed the additional finding of focal melanin pigment. This tumor showed strong and diffuse positive immunolabeling with paired box gene 8 and cytokeratin 20, and was negative with epithelial membrane antigen, carbonic anhydrase 9, CD117, cytokeratin 7, and transcription factor E3. Human melanoma black-45 showed focal positivity, but Melan-A was negative. Next-generation sequencing revealed a mutation in the TSC2 gene (c.4490C > G, p.[Pro1497Arg] and c.1257 + 1del) and break apart fluorescence in-situ hybridization with TFE3 and TFEB probes was negative. In this case report, we present the novel finding of melanin pigment occurring in a genetically proven and otherwise typical ESC-RCC, and briefly discuss the differential diagnostic considerations.

Fungal melanins have been considered as potential biosorbents due to their metal-binding properties, stability, and scalability. Previous studies established scalable fungal melanin production methods with promising strains, however, their applicability for metal-contaminated effluents treatment has not been sufficiently reported. Herein, melanin pigment derived from Amorphotheca resinae was produced and characterized using microscopy and spectroscopy techniques. Adsorptive properties towards Cu(II), Pb(II), Cd(II), and Zn(II) were evaluated using batch tests. Melanin pigment was composed of aggregates of nanosized particles with indole-based constituents. Adsorption capacities increased with the pH of solution, especially at pH > 4.0. Maximum binding capacities of Cu(II), Pb(II), Cd(II), and Zn(II) on melanin were 69.18, 103.23, 24.31, and 13.57 mg/g, respectively. The competitive adsorption experiments elucidated affinity as Cu(II)>Pb(II)≫Cd(II)>Zn(II). Adsorption time generally required <2.5 h to reach equilibrium; the pseudo-second-order kinetic model well described the kinetics. Chelating ability of free radicals in pigment was considered as a possible mechanism for adsorption. Initial adsorption capacities remained almost intact even after 5 consecutive adsorption-desorption cycles. Complete removal of Cu(II), Pb(II), and Cd(II) from metal-contaminated effluent was confirmed. Consequently, melanin pigment derived from A. resinae can be used as a biosorbent suitable for the treatment of metal-contaminated aqueous solutions.

OBJECTIVE: This study proposes a technique for visualizing the effect of facial massage using stereo-image correlation with melanin pigment.
METHODS: In this method, the melanin pigment of a subject\'s face is made visible by using an ultraviolet light and utilized as a random pattern for stereo-image correlation. Stereo-pair images of the face with the melanin pigment before and after facial massage are recorded using a desk-sized measurement equipment. Then, the deformation of the face by the massage can be obtained based on the principle of stereovision. The effectiveness of the proposed method is demonstrated by applying it to the massage effect evaluation of eight subjects (females in their 40s).
RESULTS: The results show that the massage effect can be visualized from the displacement and strain distributions across the face obtained by the proposed method. In addition, it is observed that the face is displaced significantly by the massage and individual differences between the subjects can be captured.
CONCLUSIONS: The proposed method is effective for evaluating the effect of a facial massage when the painted pattern disappears due to the applied cream during the massage.