Brown Granules Research Articles

Nuclear factor-κB p65 subunit determines the fate of aging epithelial cells

Nuclear factor (NF)-κB signaling is not only important for the immune and inflammatory responses but also for the normal development of epithelial cells, such as those in the skin and tooth. Here, we generated epithelial cell-specific p65-deficient (p65Δepi−/−) mice to analyze the roles of NF-κB signaling in epithelial cell developent. Notably, p65Δepi−/− mice exhibited no abnormalities in their appearance compared to the control (p65flox/flox) littermates. Furthermore, no major changes were observed in the skin, hair growth, and shape and color of the incisors and molars. However, 65 % of p65Δepi−/− mice exhibited corneal thickening after 8 weeks of age, and 30 % of p65Δepi−/− mice exhibited hair growth from the mandibular incisors around 24 weeks of age. No hair growth was observed at 36 and 42 weeks of age. However, micro-computed tomography images revealed a large cavity below the mandibular incisors extending to the root of the incisor. Histological analysis revealed that the cavity was occupied by a connective tissue containing hair-like structures with many dark brown granules that disappeared after melanin bleaching, confirming the presence of hair. Although inflammatory cells were also observed near the eruption site of the incisor teeth of p65Δepi−/− mice, no major disturbance was observed in the arrangement of enamel epithelial cells. Overall, these results highlight the role of p65 in the maintenance of epithelial cell homeostasis during aging.

Dermoscopic features of Melasma

Melasma, is a chronic acquired disorder of hypermelanosis of skin which is exposed to ultraviolet radiation. The clinical presentation is most often in the form of hyperpigmented patches over the face in three common patterns: Centrofacial, malar and mandibular. It is commonly seen in pigmented skin phenotypes (Fitzpatrick skin types III-V). The disease has an impact on the quality of life of patients. Dermoscopy is an in vivo noninvasive technique used to examine pigmented and amelanotic skin lesions. The technique is performed using a hand-held self-illuminating device called dermatoscope that visualizes features present under the skin surface. : To study dermoscopic (dermatoscopic) features of melasma and to distinguish between epidermal and dermal melasma based on dermoscopic features.: This study was conducted in an OPD in a clinic on 40 patients of melasma. Dermoscopy was done and their dermatoscopic features were recorded, melasma was classified as epidermal or dermal depending on dermatoscopic features. : Dermatoscopic features of melasma seen were – Accentuation of pseudoreticular pigment network, light to dark brown in colour sparing of the periappendageal region (follicular and sweat gland openings), brown granules, blue- gray perifollicular accentuation honeycomb like reticular pattern and arcuate pigmented lines. 18 patients (45%) showed epidermal type 9 patients (22.5%) revealed dermal type of melasma, 13 patients showed mixed features (epidermal and dermal). Melasma with steroid abuse showed marked erythema and telangiectasia. Dermatoscope is a valuable aid to diagnose, classify and to monitor treatment of melasma.

Study based on the acetaldehyde dehydrogenase 2 gene polymorphism and acetaminophen-induced liver injury

Objective: To explore the association between aldehyde dehydrogenase 2 (ALDH2) gene polymorphisms and abnormal liver function-induced by acetaminophen (APAP) drugs. Methods: An ALDH2 gene knockout mouse model was constructed using CRISPR/Cas9 gene editing technology. The obtained heterozygous mice were mated with opposite sex of heterozygotes. Genomic DNA was extracted from the tail of the offspring mouse. The polymerase chain reaction (PCR) method was used to determine the ALDH2 genotype. APAP was further used to induce acute drug-induced liver injury models in wild-type and ALDH2 knockout mice. Blood and liver tissues of mice were collected for liver function index, HE staining, F4/80 immunohistochemistry, and other detections. The intergroup mean was compared using a one-way ANOVA. The LSD- t test was used for pairwise comparison. Results: ALDH2 knockout mice were bred successfully. The genotyping of the offspring was segregated into the wild-type (ALDH2(+/+)), heterozygous mutant (ALDH2(+/-)), and homozygous mutant (ALDH2(-/-)), respectively. Biochemical and histological results after APAP modeling showed that the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bilirubin (TBil) was not significantly increased in the blank control group (P < 0.05), while the ALT, AST,ALP, and TBil were all elevated in the APAP experimental group. The levels of ALT (P = 0.004), AST (P = 0.002), and TBil (P = 0.012) were significantly elevated among the mutant group compared to those in the wild-type group, and the expression levels of these indicators were also significantly elevated among the homozygous mutant group compared to those in the heterozygous mutant group (P = 0.003, 0 and 0.006). In addition, the ALP levels were higher in the heterozygous mutation group than those in the homozygous mutant group (P = 0.085) and wild-type group mice, but the difference was only statistically significant compared to wild-type mice (P = 0.002). HE staining results showed that mice in the APAP experimental group had hepatocyte degeneration, necrosis, and increased inflammatory cell infiltration, which was mostly evident in mutant mice. Simultaneously, the F4/80 immunohistochemical staining results showed that brown granules were visible in the liver tissue of APAP experimental group mice, and its expression levels were significantly enhanced compared to the blank control group. Conclusion: APAP-induced liver function abnormalities were associated with the ALDH2 gene polymorphism. The liver injury symptoms were increased in ALDH2 mutant mice following APAP modeling, and the ALDH2 gene defect may alleviate, to some extent, APAP-induced liver function abnormalities.

Melanin pigmentation variations in the larval cuticle of almond moth, Ephestia cautella caused by gamma radiation

To determine the effects of gamma radiation on the melanization process and phenoloxidase activity, Ephestia cautella larvae were exposed to dosages of 200, 400, 600, 800, and 1000 Gy. After irradiation, the number of non melanized larvae and the number exhibiting a slight melanization usually increased. The degree of melanization in treated larvae differed significantly from untreated larvae. The amount of melanin usually decreases as the dosage increases and as time passes after the treatment. The results of the phenoloxidase assay indicate that the enzyme activity responds differently to radiation. For instance, at doses of 200, 400, and 800 Gy, the enzyme activity remained consistent in both control and irradiated larvae. However, at doses of 600 and 1000 Gy, the enzyme activity increased to 14.92 and 13.37 O.D. units, respectively, compared to 8.81 O.D. units in the control. In order to determine if irradiated larvae have been previously exposed to ionizing radiation, a quick and easy test based on phenoloxidase activity or the melanization response is presented for use in quarantine treatment. Histological changes, specifically in the pigment granules of melanin, were studied using a light microscope. Upon inspection of the unirradiated larvae, it was observed that brown melanin pigment granules were deposited in the epicuticle and exocuticle layers of the cuticle. When gamma radiation dosages were administered to larvae, it was observed that the melanin pigment gradually diminished until it vanished at the highest dose (1000 Gy).

Cutaneous Sarcoidosis Masquerading as Papular Amyloidosis Revealed Post--Hair Removal.

Cutaneous Sarcoidosis Masquerading as Papular Amyloidosis Revealed Post--Hair Removal.

Dermoscopy and reflection confocal microscope features of pigmented prurigo.

Pigmented prurigo (PP) is a chronic and recurrent inflammatory skin disease. PP is not common clinically, but it is easily misdiagnosed because of its diversified clinical manifestations in different stages. We retrospectively analyzed the clinical, histopathological, dermoscopy, and reflectance confocal microscopy (RCM) features of 20 patients diagnosed as PP. The female predominance ratio was revealed with male to female of 1:4. Seven female patients were on a diet (without staple food) and one patient had a history of diabetes. Eight cases were suffered in spring, six cases in winter, three cases in summer, and three cases in autumn. Multiple sites were involved in 13 cases. Four patients had urticarial papules and plaques. Nineteen patients had erythematous papules with reticular distribution, of which 14 cases accompanied reticulate hyperpigmentation, four cases with papulovesicle, and two cases accompanied with pustules. One patient only showed reticulate hyperpigmentation. In the early lesions, dermatoscopy showed pink oval lesions, punctate or linear vessels, and pale yellow rings around the skin lesions. RCM is characterized by spongiosis, spongy vesicle, neutrophils scattered in the epidermis, which was consistent with epidermis spongiosis, neutrophils infiltrating into the upper epidermis and necrotic keratinocytes in histopathology. In the fully developed lesions, dermatoscopy showed pink lesions with brown pigment granules in the center and linear vessels in the edge. RCM showed that demarcation of epidermis and dermis is not clear, and inflammatory cells can be seen in the upper dermis and histopathologically lesions assumed a patchy lichenoid pattern, and the inflammatory cells infiltrating the dermis were dominated by lymphocytes. In the late lesions, dermatoscopy showed grainy grayish-brown or yellowish-brown pigmentation surrounding the hair follicle merging with each other. RCM showed that pigment granules were increased on the ring of basal cells, inflammatory cells were sparsely infiltrated in the dermal papilla and superficial layer, and epidermis slightly hyperplastic, with melanophages and a few lymphocytes infiltrating the superficial dermis in histopathology. PP is easily misdiagnosed and not always occurs in those on a restrictive diet. A combination of dermatoscopy and RCM is helpful for its diagnosis of PP.

Histological, elemental, and ultrastructural analysis of melanin in mantle of Pacific oyster (Crassostrea gigas).

Colorful shell of bivalve is mainly because of the biological pigments, of which melanin plays an important role in shell color formation. More and more studies focus on the genes function involved in melanin synthesis, but relatively few studies address the biochemical character and ultrastructure of melanin in bivalve from microscopic perspective. Here, we investigated the histological structure of mantle of Crassostrea gigas with orange shell color. Distribution of melanin in mantle was verified with histochemical staining. In addition, immunofluorescence technique showed that strongly positive signal of CgTYR was specific to the mantle margin, which is consistence with the location of brown granules in H&E staining. The further result of elementary composition of melanin displayed that metal Ca, Fe, and Zn were detected using scanning transmission electron microscope and energy dispersive spectroscopy mapping methods. Next, based on TEM observations, it was speculated that the series of cellular events leading to the formation and release of melanin. Melanocyte in the primary stage showed many mitochondria and rough endoplasmic reticulum as well as an extensive Golgi complex with numerous vesicles intermingled with melanosome. Subsequently, melanosome was expended and their hue gradually intensified, and Golgi complex and mitochondria were still observed in the cytoplasm. Finally, after melanosome was discharged into intercellular spaces, the disintegration of membranes in some cells, and severe cellular vacuolization. These data enrich the understanding of ultrastructural characteristic and formation of melanin in mantle of bivalve and pave the way for further investigating shell coloration at the cellular level.

Rapid method of aerobic granular sludge bioreactor start-up for domestic wastewater treatment

This study presents a rapid method on how to speed up aerobic granular sludge (AGS) cultivation and ensure excellent and stable removal performance during bioreactor operation for domestic wastewater treatment. This new strategy consists of start-up the bioreactor using only anaerobic granular sludge (AnGS) as a seed and feeding with crude sewage extracted from a full-scale Extended Aeration Plant. This experiment used a 2.5 L lab-scale sequencing batch reactor (SBR). The bioreactor operated at low dissolved oxygen (DO) concentration controlled at the value of 2.0 mg/L and below. After 60 days of operation, it clearly showed that almost 90% of AnGS seeds turned from black color to brown. The physical characterization analysis showed that the average sizes were unchanged, and the granules remained compact. Also, the SBR operation monitored with brown granules showed stable removal performance. Average removal efficiencies during steady-state cycles at room temperature of COD, ammoniacal nitrogen, and phosphate reached 84%, 92%, and 100%, respectively.

Application of the Method of Melanin Bleaching After Immunohistochemical Staining of Melanin-containing Tissues.

The purpose of this research was to investigate the method of melanin bleaching in immunohistochemical staining of melanin-containing tissues. Forty melanin-containing tissue samples were selected, including 30 with malignant melanoma and 10 with blue naevi. The samples were divided into the following 3 groups: the prestaining bleaching group, nonbleaching group, and poststaining bleaching group. Each group was subjected to HMB45 and MelanA immunohistochemical staining. The detection system used was Roche ventana ultraview universal alkaline phosphatase red kit. In the prestaining bleaching group, melanin pigment was mostly removed. However, antigen expression was also affected, which resulted in significantly weaker positive staining and even represented false-negative expression. In the nonbleaching group, positive staining resulted in a red color that could easily be distinguished from the brown granules of melanin pigment. However, some melanin granules covered the tissue, impacting our observation of tissue structure. In the poststaining bleaching group, the results of positive staining were comparable to those in the nonbleaching group. The melanin pigment in these sections was almost completely removed, resulting in clearer staining and easier observation of tissue and cell structure. The method of melanin bleaching after immunohistochemical staining of melanin-containing tissue can almost completely bleach the melanin pigment and do not impact the expression of antigen. In our work, poststaining bleaching allowed for clearer immunohistochemical staining results and more obvious observation of tissue structure. This method also has the advantages of requiring simple reagents, utilizing simple operation procedures, and producing practical staining results, which makes it worthy of promotion!

Spitz Nevus (case study)

Annotation. Spitz nevus is a difficult issue in the diagnosis of skin tumors and in the clinical tactics of management of patients with this disorder. The case of diagnosis of Spitz's nevus in patient R., born in 1978 is given. in which a dermatologist's examination revealed skin tumors on the torso and extremities. Spitz's nevus was suspected. Dermatological status: on the skin of the face, neck, torso and extremities there are spots and papules, up to 6 mm (number 62) maximum, light and dark brown, without signs of inflammation, irritation and trauma. Dermatoscopic picture of pigment formations is represented by melanocyte, non-melanocyte and halo nevi. On the skin of the left breast there is a melanocyte formation asymmetric in shape in the horizontal axis, dermatoscopically corresponding to the Spitz nevus (dark brown unstructured area in the center, illuminated to the periphery with large brown granules and dots located on the periphery of the focus in the form of rays 3.8. A biopsy was performed. Histologically, there is skin with acanthosis and minor hyperortokeratosis, located in the basal layer of symmetrical, well-defined formation with refractive artifacts. The tumor is formed by vertically placed junctional nests of mainly spindle-shaped, as well as epithelioid multinucleated melanocytes with eosinophilic cytoplasm, moderate polymorphism of vesicular nuclei. The latter have numerous prominent small basophilic nucleoli. Mitotic figures and pegetoid growth are absent. Perivascular lymphocytic infiltrates are present in the papillary layer of the dermis. The patient was diagnosed with Nevis Spitz. It is recommended to monitor dynamically with dermatoscopy once a year, to avoid hyperinsolation, the use of sunscreen with SPF factor. The problem of diagnostic and treatment tactics of a patient with Spitz's nevus remains a difficult problem today. Further study of the peculiarities of Spitz's nevus in adulthood seems promising.

Melanocytes and keratinocytes morphological changes in vitiligo patients. A histological, immunohistochemical and ultrastructural analysis

ABSTRACT Vitiligo is an idiopathic acquired chronic stigmatizing disease. It is a pigmentary disorder that affects the skin and the mucous membranes, and it is characterized by well-circumscribed, depigmented milky white macules and patches. It has an estimated prevalence of 0.5–2% of the population worldwide. In the previous studies, several mechanisms such as autoimmune, oxidative stress, genetic factors, melanocytorrhagy, and neural hypothesis have been suggested for vitiligo pathogenesis.We aimed to assess the morphological changes of epidermal melanocytes and keratinocytes in patients with vitiligo. This aim will be fulfilled by histological, ultrastructural, and immunohistochemical analysis of skin biopsies from lesioned and non-lesioned sites in vitiligo patients.The study was carried out on 15 selected patients with stable vitiligo vulgaris but not receiving treatment in the last year and they fulfilled our inclusion criteria.Biopsies were taken from lesioned and non-lesioned sites in the same vitiligo patients, and they are processed for examinations by LM (using Hx & E, and Masson Fontana stain), immunohistochemical analysis (using Melan-A, E-cadherin, and caspase-3), and TEM (to demonstrate the ultra-structures).By LM, staining with Hx & E, lesioned skin in vitiligo patients showed hyperkeratosis, basal vacuolization, acanthosis with an increase in the epidermal thickness, ballooning of keratinocytes, and spongiosis. Regarding melanocytes, we observed a few numbers of melanocytes, also we detected some basal epidermal cells contain brown melanin granules. Using Fontana-Masson stain, we found that the melanin pigment is present in both lesioned and non-lesioned skin of vitiligo patients. We confirmed the presence of melanocytes in the lesioned skin by the immunohistochemical staining with Melan-A. The epidermal cells in lesioned skin of vitiligo patients showed weak positive expression of E-cadherin between them and an increase in the number of apoptotic Caspase-3 positive cells. BY TEM, the lesioned skin in vitiligo patients showed that the keratinocytes and melanocytes had various degenerative changes, disturbance of desmosomes in between keratinocytes, and absence of melanosomes in the keratinocytes. The detected melanocytes were degenerated and contained some melanosomes, melanin granules, and autophagosomes.We concluded that vitiligo pathogenesis is a combination of several factors and cannot be explained by only one mechanism. The pathology in the lesioned vitiliginous skin is a combination of several degenerative changes in keratinocytes, and melanocytes.

Two Novel Species and Two New Records within the Genus Pluteus (Agaricomycetes, Agaricales) from Thailand

Pluteus is a species-rich genus of saprotrophic agaric in the family Pluteaceae and is widely distributed in tropical, subtropical, and temperate areas throughout the world. Some species in this genus are threatened species according to the International Union for Conservation of Nature (IUCN) red list. During investigations of agarics in northern Thailand, four Pluteus taxa were collected. Morphological characteristics and phylogenic analyses were investigated. Two new species, namely P. chandrasikuliae and P. saisamorniae, were introduced. Pluteus chandrasikuliae is characterized by its relatively large basidiomata, an applanate, dark brown scaly pileus with a cutis pileipellis, two types of hymenial cystidia viz. irregular, as well as diverticulate cells and lageniform cells. In accordance with the phylogenetic results, this new species belongs to the Pluteus sect. Celluloderma. Moreover, P. saisamorniae is distinguished by a plano-convex with a broad umbo, dark brown minute squamules pileus, light brown lamellar edges, greyish orange stipe covered with brown granules, subglobose to broadly ellipsoid basidiospores, an abundance of thin- to thick-walled cheilocystidia and pleurocystidia, a trichohymeniderm pileipellis, and brown caulocystidia in clusters. Pluteus saisamorniae is a member of Pluteus sect. Hispidoderma. Additionally, P. losulus and P. septocystidiatus were discovered in Thailand for the first time and they belong to Pluteus sect. Pluteus. Comprehensive descriptions along with illustrations, photographs, phylogenetic trees showing their positions, and a comparison with phenetically similar taxa are provided.

The expression pattern of CD10 and CD31 identifies fine fibrovascular stroma of grade 1-endometrial endometrioid carcinomas in cytology.

The objective of this study was to assess the diagnostic utility of CD10 in the differential diagnosis of grade 1-endometrial endometrioid carcinoma (G1-EEC) and the metaplastic changes associated with the endometrial glandular and stromal breakdown (EGBD) on liquid-based cytological (LBC) samples. (1) The type and distribution of CD10-positive cells in EGBD and G1-EEC patients were evaluated. (2) Based on the results from (1), histological and cytological specimens were double-immunostained with CD31 and CD10 to confirm whether CD10-positive tubular-canalicular material found in (1) was represented by fine threads of endometrial-type fibrovascular stroma. (3) Based on the results from (2), additional immunostaining of histological specimens was performed for CD146 and αSMA as markers of perivascular cells. (1) CD10 positive cells showed two main patterns of expression: cytoplasmic immunoreactivity in the form of dense brown granules in EGBD and tubular-canalicular branching patterns in G1-EEC. (2) The tubular-canalicular material observed in cytological specimens of G1-EEC samples co-expressed CD10 and CD31, and was interpreted as representing fine threads of endometrial fibrovascular stroma in the corresponding histological samples. Conversely, metaplastic changes in EGBD cases, only a few CD31-positive signals were found inside the condensed stromal clusters with CD10-positive. (3) Cells surrounding the CD31-positive vascular endothelial cells expressed CD146 and αSMA; moreover, some of the thin CD10-positive fibrous stromal strands also co-expressed αSMA. CD10 is a very useful immunomarker for distinguishing between G1-EEC and the metaplastic changes of EGBD in LBC samples.

Exploration of mechanisms for calcium phosphate precipitation and accumulation in nitrifying granules by investigating the size effects of granules

Calcium phosphate could be accumulated in aerobic granules, which attracts attention recently for phosphorus removal and recovery from wastewater. In this study, partial nitrifying granules with high calcium precipitate content were sorted into different size groups for characterization and evaluation to reveal the dynamic balance of granules at stead state and relevant calcium phosphate precipitation and accumulation mechanism. It was found that light yellow small granules without calcium precipitates but high microbial activity co-existed with deep brown granules with calcium precipitate of around 91% and low microbial activity. Characterization with specific oxygen uptake rates, specific ammonium oxidation rates, calcium and phosphate removal rates from solution, EPS contents, elemental compositions by energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and scanning electron microscopy (SEM) for different size groups of granules enabled a proposal of a new hypothesized mechanism for calcium precipitation and accumulation. With this proposed mechanism, it is believed that sufficient granule retention time in reactors was critical for the accumulation of calcium precipitates followed by a slow microbial growth rate of biomass due to mass transfer resistance. The co-precipitation of calcium carbonate and calcium phosphate mainly occurred in granules with a size less than 710 µm while calcium phosphate dominant minerals were accumulated in granules larger than 710 µm. The results and conclusions in this study shed light on the mechanisms of calcium phosphate accumulation in granules, which could be used to better operate and control aerobic granular sludge with calcium phosphates for phosphorus removal and recovery.

Leptin expression is substantially correlated with prognosis of urinary bladder carcinoma

ABSTRACT This study examined leptin expression in cases of bladder cancer and its diagnostic and prognostic usefulness in bladder malignancies. A set of 128 urinary bladder cancer cases and 24 normal specimens of bladders were employed for an immunohistochemical investigation of leptin expression in tissue microarrays. Leptin was up-regulated during transformation and was identified as brown cytoplasmic granules in the malignant urothelium of 123 (96%) bladder neoplasms, of which 68 (53.1%) cases showed high levels (moderate to strong) of staining. Strong staining was found to be associated with high stages (P = 0.001), muscularis propria infiltration (P < 0.001), vascular invasion (P < 0.03), lymph node involvement (P < 0.02), metastases (P < 0.05), and mortality (P < 0.03). Furthermore, various important survival distributions were detected with leptin expression in the malignant urothelium (P < 0.03). These pilot results suggest that leptin might be a valid marker for predicting the stage and bad prognoses in bladder carcinoma.

A 90-day Sub-chronic Oral Toxicity Assessment of Mulberry Extract in Sprague Dawley Rats.

Mulberry extract from Fructus Mori contains an anthocyanin pigment and has been widely used as a food additive in China and other Eastern Asian countries. Only few research has been done on toxicological profiling of mulberry extract for its safety evaluation; however, the data is inconclusive. In the current study, mulberry extract of 4200, 1400, or 466 mg/kg were orally administrated to Sprague Dawley rats for 90 consecutive days followed by a recovery period of 28 days. No abnormalities were detected in body weights, food intake, ophthalmological, hematological, coagulation, clinical chemistry, and organ weights parameters. Discoloration of urine (red, purple, and brown) and feces (black), along with bedding material (purple) were observed in the 4200 mg/kg group. Further, microscopic examination revealed brown granules in the renal tubular cells for rats in 4200 and 1400 mg/kg groups. Since these changes were associated with excretory effect of the extract, the No Observed Adverse Effect Level was determined to be 4200 mg/kg, which was equivalent to the 1058.5 mg/kg of anthocyanin.

SIMPLIFIED MYELOPEROXIDASE STAIN USING BENZIDINE DIHYDROCHLORIDE.

THE DEMONSTRATION of leukocyte peroxidase activity has been of practical value to the hematologist since the beginning of the present century. Many technics have been described based on the use of benzidine base as an indicator. 1. , 2. , 3. , 4. , 5. , 6. , 7. This compound when oxidized precipitates as brown or blue insoluble crystals or granules.

The efficacy and safety of topical tranexamic acid (liposomal or lotion with microneedling) versus conventional hydroquinone in the treatment of melasma.

Robust evidence regarding the efficacy of topical tranexamic acid (TA) on melasma in Chinese population is lacking. To evaluate the efficacy and safety of 1.8% liposomal TA and microneedling with 5% TA solution on melasma. Sixty melasma patients were enrolled and randomized to receive 1.8% liposomal TA twice daily, microneedling with 5% TA solution weekly or 2% hydroquinone every night. Objective and subjective assessments were obtained at baseline, 4, 8, and 12weeks. 27.8% of patients of liposomal TA group, 33.3% of microneedling with TA solution group, and 30.0% of hydroquinone group were recognized as "more than 50% improvement." At the endpoint, the melanin index (MI) in all treatment groups was significantly decreased, while the improvement of MI in microneedling with TA solution group and hydroquinone group is higher than liposomal TA group. The erythema index (EI) was significantly diminished in liposomal TA group and microneedling with TA solution group. Dermatoscopy and reflectance confocal microscopy revealed decreased brown granules in all groups and reduced telangiectasia in liposomal TA group and microneedling with TA solution group. 1.8% liposomal TA and microneedling with 5% TA solution are both effective and safe on melasma.

S1774 Primary Anorectal Malignant Melanoma: Rare and Fatal

INTRODUCTION: We present an uncommon diagnosis of primary anorectal malignant melanoma (ARMM) in a patient with anemia and altered bowel habits. Primary ARMM accounts for less than 4% of anorectal malignancies and less than 1.5% of all melanomas. ARMM can often go undiagnosed, which is detrimental to patients due to its aggressive nature. CASE DESCRIPTION/METHODS: 63 y.o. African American male with a history of ascending colon resection for colon adenocarcinoma 8 years prior was referred for change in bowel habits and nausea with eating. He had worsening intermittent constipation, but denied melena or blood in his stool. He had nausea 45 minutes after eating but no vomiting. He denied recent weight loss. His last colonoscopy was a year after colon resection. The ileocolonic anastomosis was at 70 cm from anal verge, there was a 3 mm hyperplastic polyp in the sigmoid and repeat colonoscopy was recommended in a year. Physical exam was negative for skin hyperpigmentation, moles or lesions. Abdominal exam was negative for tenderness, guarding or rebound. Labs showed normocytic anemia, Hgb 8.1 g/dl from 12 g/dl the year prior. Digital rectal exam revealed a firm immobile mass, 3 cm from anal verge. Colonoscopy showed a fungating, ulcerated, non-obstructing 3 cm mass in the rectum. Biopsies were done with cold forceps for histology. Pathology noted the mass had brownish pigmentation and extensive necrosis, consistent with malignant melanoma. Immunohistochemical staining was positive for S-100, HMB-45, Melan-A and SOX-10 supporting the diagnosis. A diagnosis of primary anorectal malignant melanoma was made as patient had no evidence of cutaneous or ocular melanoma. PET scan showed metastasis to a large pelvic lymph node so patient was not a surgical candidate. Oncology started patient on immunotherapy with ipilmumab and nivolumab. DISCUSSION: Primary ARMM is a rare malignancy, especially in African Americans such as our patient, with a poor prognosis. As with our case, presenting symptoms can be unspecific such as anemia. Often patients with ARMM present with rectal bleeding that is attributed to hemorrhoids and endoscopic intervention is not pursued. This is detrimental as it causes a delay in diagnosis. Our patient’s mass was pigmented, but in 29% of cases the mass may be amelanotic resulting in misdiagnosis. Treatment is abdominoperineal resection if there is no evidence of metastasis. Our case highlights the importance of recognizing primary ARMM as a cause of rectal bleeding in patients with an anorectal mass.Figure 1.: Retroflexed view in the rectum revealing the pigmented, ulcerated, non-obstructing 3 cm mass at 3 cm from anal verge.Figure 2.: The mass is composed of melanoma cells that formed irregular nests and cords, and manifested enlarged and hyperchromatic nuclei with irregular nuclear contours and prominent nucleoli. The pigments were finely granular and brownish. (H&E, x200).Figure 3.: Immunohistochemistry demonstrated the tumor cells to be positive for S-100 (Fig. A, cytoplasmic stain, fast red; X100), HMB-45 (Fig. B, cytoplasmic stain, fast red; X100) and SOX-10 (Fig. D, nuclear stain, fast red; X100). They were negative for expression of pan-cytokeratin, the brown granules in the background are melanin pigments (Fig.C, AE1/3, lack of cytoplasmic stain, DAB; X100).

The Correlate between Argyrophilic Nucleolar Organizer Region Stain and Ki-67 Immnochistochemistry in Diagnostic Breast Cancer

Introduction: This experimental study gauged the value of argyrophilic nucleolar organizer region (AgNOR) staining as a possible technique for the estimation of cell kinetics in conventional histology sections, in benign and malignant breast lesions. Methods: With a silver staining technique and immunohistochemistry, we associated the numbers of AgNORs and Ki67 scores in 30 breast carcinomas and 10 benign breast lesions. Results: The mean values of Ag NORs silver stain dots count for normal, benign, grade II and III were 1.28±0.17, 2.83±0.68, 5.23±0.87 and 7.32±0.92, respectively. There was a statistically significant difference (p≤0.001) was noticed between the all individual groups, among the normal and breast lesion as well as among the GII and GIII. Immunohistochemical Results of Ki-67 protein exhibited homogenous golden-brown color in control case and a positive brown granules or diffuse dark brown color in the nuclei of both benign and malignant cases under the 400X magnify examined under the light microscope. Discussion: AgNOR counts performed on routine formalin-fixed paraffin sections could provide substantial kinetic evidence. Additionally, the difference in AgNOR counts between benign and malignant tumors is such that they may be of diagnostic worth.