Dysplastic Lesions Research Articles

Application of immunohistochemical staining of PRAME in differential diagnosis between melanoma and melanocytic nevus

Objective: To investigate the diagnostic value of preferentially expressed antigen in melanoma (PRAME) in differential diagnosis of benign and malignant cutaneous melanocytic lesions. Methods: Fifty-nine cases of melanoma (50 cases of skin primary melanoma, and 9 cases of metastatic melanoma) and 48 cases of melanocytic nevus (40 cases of common nevus and 8 cases of dysplastic nevus) were subject to PRAME immunohistochemistry staining.The difference of PRAME expression between melanoma and melanocytic nevus was analyzed. Results: Among the 50 patients with primary cutaneous melanoma, there were 23 males and 27 females ranging in age from 33 to 87 years (average age 62.4 years, median age 64.5 years). Among the 9 metastatic melanoma there were 7 males and 2 females ranging in age from 40 to 82 years (average age 64 years, median age 65 years). Twenty-six cases (26/50, 52.0%) of cutaneous primary melanoma and 4 cases (4/9) of metastatic melanoma showed diffuse positive PRAME staining. 40 cases (40/40, 100%) of common nevus and 8 (8/8) cases of dysplastic nevus were PRAME negative. Compared with melanocytic nevus group, the melanoma group included more cases with diffuse positive PRAME staining (P<0.05). The sensitivity and specificity of using PRAME to differentiate primary cutaneous melanoma from melanocytic nevus in the cohort is 52.0% and 100%. Conclusions: There is a significant difference in the expression of PRAME between melanoma and melanocytic nevus.Thus, PRAME can be used as an auxiliary diagnostic tool for differentiating benign from malignant cutaneous lesions.

Fractal Dimension Analysis of Melanocytic Nevi and Melanomas in Normal and Polarized Light-A Preliminary Report.

Clinical diagnosis of pigmented lesions can be a challenge in everyday practice. Benign and dysplastic nevi and melanomas may have similar clinical presentations, but completely different prognoses. Fractal dimensions of shape and texture can describe the complexity of the pigmented lesion structure. This study aims to apply fractal dimension analysis to differentiate melanomas, dysplastic nevi, and benign nevi in polarized and non-polarized light. A total of 87 Eighty-four patients with 97 lesions were included in this study. All examined lesions were photographed under polarized and non-polarized light, surgically removed, and examined by a histopathologist to establish the correct diagnosis. The obtained images were then processed and analyzed. Area, perimeter, and fractal dimensions of shape and texture were calculated for all the lesions under polarized and non-polarized light. The fractal dimension of shape in polarized light enables differentiating melanomas, dysplastic nevi, and benign nevi. It also makes it possible to distinguish melanomas from benign and dysplastic nevi under non-polarized light. The fractal dimension of texture allows distinguishing melanomas from benign and dysplastic nevi under polarized light. All examined parameters of shape and texture can be used for developing an automatic computer-aided diagnosis system. Polarized light is superior to non-polarized light for imaging texture details.

DNA Fragmentation and mRNA Expression of Bcl-2, Bcl-xL, p53, p21 and HSP70 Genes in Nondysplastic and Dysplastic Oral Lichen Planus.

Background:Oral lichen planus (OLP) is a chronic inflammatory disease. Apoptosis of the basal keratinocytes is a causative factor for OLP pathogenesis but the detailed mechanism of apoptosis among nondysplastic and dysplastic OLP lesions is yet unraveled.Aims:This study aims to evaluate the involvement of cellular DNA fragmentation and alteration in the expression of Bcl-2 and B-cell lymphoma extra-large (Bcl-xL), p53, p21 and heat shock protein 70 (HSP70) in nondysplastic and dysplastic OLP lesions.Materials and Methods:Untreated, fifteen OLP patients each with nondysplastic and dysplastic lesions were enrolled for this study. Their DNA fragmentation was analyzed by the agarose gel electrophoresis method. The mRNA expression of Bcl-2, Bcl-xL, p53, p21 and HSP70 were measured using semi-quantitative reverse transcription-polymerase chain reaction.Results:Elevated DNA fragmentations were found in dysplastic lesions compared to nondysplastic type. Significantly higher expression of Bcl-2, Bcl-xL, p53 and p21 were found in both types of OLP lesion compared to the control. Expression of Bcl-2 and Bcl-xL were significantly elevated in nondysplastic lesions, whereas significantly overexpression of p53 and p21 were found in dysplastic lesions. Anti-stress protein HSP70 was overtly expressed in dysplastic lesions compared to other groups.Conclusion:Reduced expression of Bcl-2 and Bcl-xL, with elevated DNA fragmentation, may be associated with increased apoptosis in dysplastic lesions which aid in the resolution of the chronic inflammatory process. Higher expression of p53 and p21 in dysplastic lesions reflect its malignant potentiality. Overexpression of HSP70 in dysplastic lesions is a useful marker for higher cellular stress.

Recognition of Skin Lesions in Dermoscopic Images using Local Binary Patterns and Multinomial Logistic Regression

Skin cancer, such as melanoma, affects more than 13 million people annually and causes more than 65,000 deaths. As with almost all types of cancer, it can be treated effectively if it's diagnosed in it's early stages. The development of computer-aided diagnostic tools is, therefore, a valuable and urgent need to suport the decision of dermatologists, minimize human error, and accelerate the time to diagnosis and the prognosis of patients. The present work proposes the automatic recognition of three skin lesions, namely Common Nevus, Atypical Nevus, and Melanoma, from dermoscopic images. First, a feature vector representing texture is extracted from the dermoscopic images using the local binary pattern operator (LBP). Then, a multinomial logistic regression (MRL) model is used to exploit the feature vector to perform the multiclass classification. The hyperparameters of the LBP operator, namely the radius and the pixel neighborhoods, were modified to provide the feature set to the MRL model that maximizes the classification performance. Moreover, to assure the generalizability of the model and the reproducibility of the results, a 10-iteration Monte Carlo cross-validation (MCCV) was employed, by dividing the dataset into 70% for training and 30% for testing, with different random seeds in each iteration. The optimal LBP-MRL classification model was finally embeddeb in a graphical user interface (GUI) to facilitate the interaction between the user and the classification system. An average accuracy of 90%, averge recall of 89.23%, 84.75% and 100%, and average precision of 89.95%, 88.12% and 100% were obtained, to identify Common Nevus, Atypical Nevus and Melanoma, respectively, in the MCCV strategy. It should be highlighted that the proposed classification system can perfectly differentiate melanoma from the other two skin lesions.

Increased Risk of Non-conventional and Invisible Dysplasias in Patients with Primary Sclerosing Cholangitis and Inflammatory Bowel Disease.

Patients with primary sclerosing cholangitis and inflammatory bowel disease [termed PSC-IBD] have a higher risk of developing colorectal neoplasia than those with IBD alone. The mechanism by which concomitant PSC increases the risk of colorectal neoplasia remains unknown. Seven distinct non-conventional dysplastic subtypes have been recently described in IBD, including crypt cell dysplasia, hypermucinous dysplasia, goblet cell-deficient dysplasia, dysplasia with increased Paneth cell differentiation [DPD], sessile serrated lesion [SSL]-like dysplasia, traditional serrated adenoma [TSA]-like dysplasia, and serrated dysplasia, not otherwise specified [NOS]. Despite the lack of high-grade morphological features, crypt cell, hypermucinous, and goblet cell-deficient dysplasias often show molecular features characteristic of advanced neoplasia [i.e. aneuploidy and KRAS mutations] and are more frequently associated with advanced neoplasia than conventional dysplasia on follow-up. We aimed to characterise clinicopathological features of dysplasia found in PSC-IBD patients. A cohort of 173 PSC-IBD patients were analysed. All dysplastic lesions were subtyped as either conventional or non-conventional dysplasia. The clinicopathological features of PSC-IBD patients with neoplasia were also compared with those of non-PSC IBD patients with neoplasia. There were 109 [63%] men and 64 [37%] women, with a mean age of 26 years at IBD diagnosis and a long history of IBD [mean duration: 14 years]. Ulcerative colitis was the most common IBD subtype [80%], and the majority of patients [92%] had a history of pancolitis. A total of 153 dysplastic lesions were detected in 54 [31%] patients, 35 [65%] of whom had multifocal dysplasia. One additional patient presented with colorectal cancer [CRC] without a history of dysplasia. Dysplasia was often non-conventional [n = 93; 61%], endoscopically/grossly invisible [n = 101; 66%], and right/proximal-sided [n = 90; 59%]. All seven non-conventional subtypes were identified, including 46 [30%] crypt cell dysplasia, 23 [15%] hypermucinous dysplasia, 12 [8%] goblet cell-deficient dysplasia, seven [5%] DPD, three [2%] TSA-like dysplasia, one [1%] SSL-like dysplasia, and one [1%] serrated dysplasia NOS. Follow-up information was available for 86 lesions, of which 32 [37%] were associated with subsequent detection of advanced neoplasia [high-grade dysplasia or CRC] within a mean follow-up time of 55 months. PSC-IBD patients with neoplasia were more likely to have pancolitis [98%, p = 0.039] and a longer IBD duration [mean: 17 years, p = 0.021] than those without neoplasia [89% and 12 years, respectively]. When compared with a cohort of non-PSC IBD patients with neoplasia, the PSC-IBD group with neoplasia was more often associated with non-conventional [61%, p <0.001], invisible [66%, p <0.001], and right/proximal-sided [59%, p = 0.045] dysplasias [vs 25%, 21%, and 47%, respectively, for the non-PSC IBD group]. The rate of advanced neoplasia was nearly 2-fold higher in the PSC-IBD group [37%] compared with the non-PSC IBD group [22%] [p = 0.035]. Nearly a third of PSC-IBD patients developed dysplasia, which is often associated with non-conventional dysplastic features, invisible endoscopic/gross appearance, right/proximal-sided colon, multifocality, and advanced neoplasia on follow-up. These findings underscore the importance of recognising these non-conventional subtypes by practising pathologists and the need for careful and frequent endoscopic surveillance, with random biopsies, in PSC-IBD patients.

Detection of Human Papillomavirus in Squamous Lesions of the Conjunctiva Using RNA and DNA In-Situ Hybridization.

In-situ hybridization provides a convenient and reliable method to detect human papillomavirus (HPV) infection in formalin-fixed paraffin-embedded tissue. Cases of conjunctival papillomas, conjunctival intraepithelial neoplasia (CIN), conjunctival carcinoma in situ (cCIS), and invasive squamous cell carcinoma (SCC), in which low-risk (LR) and/or high-risk (HR) HPV types were evaluated by RNA or DNA in-situ hybridization, were retrospectively identified. LR HPV types were frequently detected in conjunctival papillomas (25/30, 83%), including 17/18 (94%) with RNA probes, compared to 8/12 (75%) with DNA probes. None of the CIN/cCIS or SCC cases were positive for LR HPV by either method. HR HPV was detected by RNA in-situ hybridization in 1/16 (6%) of CIN/cCIS cases and 2/4 (50%) of SCC cases, while DNA in-situ hybridization failed to detect HPV infection in any of the CIN/cCIS lesions. Reactive atypia and dysplasia observed in papillomas was generally associated with the detection of LR HPV types. Collectively, our findings indicate RNA in-situ hybridization may provide a high-sensitivity approach for identifying HPV infection in squamous lesions of the conjunctiva and facilitate the distinction between reactive atypia and true dysplasia. There was no clear association between HPV infection and atopy in papillomas or dysplastic lesions.

Acetic acid versus toluidine blue as screening tools for oral potentially malignant disorders.

Diagnostic adjuncts such as toluidine blue have been investigated as screening tools that improve visual examination of potentially malignant disorders (PMD) and oral cancer. Acetic acid has been reported to be of value in the early detection of cervical cancers. This study assessed the utility of 5% acetic acid as a diagnostic adjunct in oral PMD and compared the accuracy of acetic acid with toluidine blue in the detection of dysplastic PMD and high-risk lesions. This cross-sectional study was conducted at a dental hospital in a rural setting. Thirty-one patients with oral PMD formed the study group. Five percent acetic acid was applied to the lesions, followed by toluidine blue application and biopsy. Sensitivity, specificity, and positive and negative predictive values were computed considering true positives as stain uptake in dysplastic and high-risk PMD. The sensitivity, specificity, and positive and negative predictive values of acetic acid for identifying dysplastic or malignant lesions were 100%, 13.3%, 51.2%, and 100%, respectively, and that for toluidine blue were 75%, 100%, 100%, and 78.9%, respectively. The corresponding values for identifying high-risk PMD (lesions with moderate and severe dysplasia) using acetic acid were 100%, 9.1%, 25.9%, and 100%, respectively, and for toluidine blue were 85.7%, 81.8%, 60%, and 94.7%, respectively. The utility of acetic acid in detecting dysplasia and high-risk PMD is severely limited due to its poor specificity. Compared with acetic acid, toluidine blue is a superior screening tool.

Reproducibility of the histopathologic diagnosis of melanoma and related melanocytic lesions: Results from a testing study and a reference guide for providers

To the Editor: The alarming increase in melanoma incidence demands the evaluation of biases in screening and histologic diagnosis.1 Absent disease outcome data, no gold standard exists for the accuracy of histopathology.2 We thus examined intraobserver reproducibility among board-certified or fellowship-trained dermatopathologists, the study design we believed a priori would capture the highest experimental reproducibility rates for melanocytic lesion diagnosis. We focused on the conceptual “common” pathway of melanomagenesis, namely nevus, dysplastic nevus, melanoma in situ (MIS), and invasive melanoma, reflecting low cumulative solar damage and representing the most frequently biopsied (∼80%) melanocytic lesions.

FHER2, PR, ER, Ki-67 and Cytokeratin 5/6 Expression in Benign Feline Mammary Lesions.

Simple SummaryFeline mammary neoplasias are highly prevalent in domestic cats and present many similarities to their human counterparts. Since information about benign feline mammary lesions is still scarce and often controversial, studies using a wider panel of oncological biomarkers are necessary to understand their potential contribution to malignant lesions. This study analyzed 47 benign lesions from 27 queens, regarding the expression of estrogen and progesterone receptors (ER and PR, respectively), fHER2 protein and two malignancy indicators (Ki-67 and CK 5/6). Our results showed that most of the lesions were ER positive (91.5%), PR negative (63.8%), fHER2 negative (64.4%), Ck 5/6 negative (76.6%) and with a low Ki-67 index (78.7%). Additionally, significant correlations were found between younger ages and ER positivity and between larger lesions and negative PR status. Our results highlight the importance of estrogen receptors in the development of benign feline mammary lesions, further contributing to the development of preventive and monitoring strategies in feline mammary oncology.Biomarkers are essential in the characterization of neoplastic lesions and aid not only in the classification of the nature of the lesions, but also in the understanding of their ontogeny, development and prognosis. In cats, while mammary carcinomas are increasingly being characterized, information on their benign lesions is still scarce. Indeed, a better characterization of benign lesions could have an important role in unravelling mammary oncogenesis, similar to that in human breast cancer. Thus, in this study, the expression of five markers was analyzed in 47 benign mammary lesions (hyperplasia, dysplasia and benign tumors) collected from 27 queens. Dysplastic and hyperplastic lesions were the most common (41/47, 81.7%). Most of the lesions were classified as ER positive (43/47, 91.5%), PR negative (30/47, 63.8%), fHER2 negative (29/47, 64.4%), CK 5/6 negative (36/47, 76.6%) and with a low Ki-67 index (37/47, 78.7%). Statistical analysis revealed a correlation between younger ages and ER positivity (p = 0.013) and between larger lesions and negative PR status (p = 0.038). These results reinforce the importance of evaluating the expression of the ER status, prevalent in benign lesions, as a putative precursor in cancer progression.

Recognizing Histopathological Simulators of Melanoma to Avoid Misdiagnosis.

Melanocytic lesions have a wide morphological spectrum, ranging from benign nevi to malignant melanoma. In contrast to a diagnosis of a benign nevus, a diagnosis of melanoma could mean intensive treatment, lifetime monitoring, and a worse prognosis. Therefore, melanocytic tumors are notoriously challenging and associated with a high risk of litigation in surgical pathology. After describing the basic features of nevi and melanoma, this article describes the detailed clinical and histological features of those lesions that share many similar features with melanoma. The entities included are Spitz nevi and atypical Spitz tumors (AST), Reed nevus, dysplastic nevus, cellular blue nevus (CBN), deep penetrating nevus, combined nevus, recurrent nevus, irritated nevus, congenital pattern nevus, acral nevus, and nevi of special sites. Knowledge of these imitators can help pathologists distinguish between benign and malignant cases and avoid misdiagnosis.

Abstract 5345: Novel approach to attenuate melanoma initiation and progression

Abstract Background and Purpose: Melanocytic nevi are benign proliferations of melanocytic cells that are positively correlated with susceptibility to melanoma. Polyaromatic hydrocarbons (PAH) are ubiquitous environmental pollutants that are potent mutagens and carcinogens, and the investigators have taken advantage of these properties to investigate the mechanisms by which chemicals cause cancer of the skin and other organs. In comparison to C57Bl/6 strain, C3H/HeN mice are more susceptible to the development of PAH induced tumors. Therefore, a mouse model of nevus initiation and progression was developed in C3H/HeN mice using a modified chemical carcinogenesis protocol. Experimental Design: In the traditional two-stage skin carcinogenesis model, initiation is accomplished by the application of a subcarcinogenic dose of a carcinogen. Subsequently, tumor development is elicited by repeated treatment with a tumor promoting agent. Nevi develop due to DNA damage initiated by 7, 12-dimethylbenz[a]anthracene (DMBA) followed by chronic promotion with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Results: Dysplastic pigmented skin lesions appeared in ~10 wk with 100% penetrance. Studies reported here demonstrating that inhibition or depletion of RLIP results in apoptosis and cancer regression in an exceptionally broad spectrum on neoplasia have suggested an intriguing possibility that RLIP is an anti-apoptotic mechanism required not only for the survival of cancer cells, but also for their very existence. In support of this assertion results presented here that the well-known carcinogens, DMBA and phorbol esters (PMA or TPA) are ineffective in causing neoplasia in C3H/HeN mice treated with RLIP-targeting agent’s (RLIP antibodies and RLIP antisense). Here we demonstrate that DMBA/PMA-induced skin carcinogenesis in C3H/HeN mouse was suppressed completely by depletion of RLIP by antisense or inhibition by antibodies. In addition, C3H/HeN mice treated with RLIP-targeting agent’s, p53, P38, and JNK activation did not occur in response to carcinogens. These findings demonstrate a fundamental role of RLIP in chemical carcinogenesis. Conclusions: We demonstrate that RLIP expression is significantly greater in cancer cells than in non-malignant cells, and RLIP-targeting agent’s (RLIP antibodies and RLIP antisense) treatment was significantly suppress the generation of melanocytic nevi and their progression to melanoma. (This work was supported in part by the Department of Defense grants W81XWH-16-1-0641 and W81XWH-20-1-0362. Funding from the Beckman Research Institute of City of Hope is also acknowledged). Citation Format: Sharad S. Singhal, Prakash Kulkarni, Jyotsana Singhal, David Horne, Sanjay Awasthi, Ravi Salgia. Novel approach to attenuate melanoma initiation and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5345.

Abstract 1553: miR-1 targets Cdk6 and controls cell cycle progression and apoptosis in colitis-associated dysplasia

Abstract Detection of colorectal dysplasia during surveillance colonoscopy is currently the best method of determining risk of colitis-associated colorectal cancer (CAC). An understanding of the early molecular changes associated with the development of these lesions will inform the identification of new biomarkers for earlier detection. miRNAs (miRs), highly conserved noncoding RNAs, show great promise as stable, tissue-specific biomarkers of neoplasia. We previously identified 12 miRs that are differentially-expressed in colitis-associated dysplasias (flat and polypoid) vs. inflamed colonic mucosa from mice treated with AOM/DSS. miR-1, a putative tumor suppressor, was downregulated in colitis-associated dysplasias. Analysis of the mRNA expression profile of AOM/DSS-induced dysplasias and prediction of the interactions between the miRs and their targets led to the selection of the Cdk6 as the target of miR-1 to be further investigated, based on its: 1) upregulation in AOM/DSS-induced dysplasia; and 2) association with cell cycle progression and inflammatory signaling. The goal of the present study was to validate the predicted interaction between miR-1 and Cdk6 and assess the biological function of miR-1 in vitro. The ability of miR-1 to interact with the 3’UTR of Cdk6 mRNA was assessed using a dual luciferase assay. Co-transfection of HCT116 or RKO colon carcinoma cells with Cdk6-WT and miR-1 mimics led to a significant reduction in relative luciferase activity in both cell lines (30%, p=0.0117 and 47%, p=0.0269; respectively). Transfection with the Cdk6-Mut did not alter relative luciferase activity, confirming the Cdk6 binding site was specific for miR-1. The biological function of miR-1 was assessed in HCT116 and RKO cells reverse-transfected with miR-1 and cel-miR-67 (negative control) for 48 hrs. Apoptosis (AnnexinV+ cells) and cell cycle progression (% of cells in G0/G1, G2/M and S phase) were evaluated by flow cytometry, and proliferation by cell count (Trypan Blue). HCT116 and RKO transfected with the miR-1 mimic exhibited a higher proportion of apoptotic cells than the negative control (30% and 20% increase, p=0.0022 and 0.0013, respectively). Cell cycle analyses revealed the miR-1 mimic induced cell cycle arrest (G0/G1) in both cell lines (p&amp;lt;0.05). In HCT116 cells, this was accompanied by a reduction in the percentage of cells in G2/M (p=0.002) and S (p&amp;lt;0.001) phase. No effect of miR-1 on total cell number was observed. These results demonstrate that Cdk6 is a direct target of miR-1, and suggest that downregulation of this miR in dysplastic lesions contributes to CAC by inducing cell cycle progression and inhibiting apoptosis. These data provide novel insight into the early molecular changes that accompany the development of colitis-associated dysplasia and may serve as biomarkers for early detection of neoplasia. Supported by the Timothy P. and Aurora M. Hughes Fund for Colon Cancer Research. Citation Format: Mariana F. Fragoso, Geysson J. Fernandez, Lisa Vanderveer, Harry S. Cooper, Michael Slifker, Margie L. Clapper. miR-1 targets Cdk6 and controls cell cycle progression and apoptosis in colitis-associated dysplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1553.

Incidental Gallbladder Neoplasms: A Growing Global Burden.

BackgroundThe increasing trend of laparoscopic procedures has made cholecystectomies one of the most common surgical specimens received for histopathological evaluation. This has also led to an increasing trend of finding incidental gallbladder malignancies for a presumed benign disease. The present study describes the histopathological spectrum of neoplastic lesions of the gallbladder along with the historadiological correlation with special emphasis on incidental gallbladder carcinomas (IGBC).Materials and methodsAll the cholecystectomies received over a span of two and a half years were studied. Demographic details, imaging findings, gross features, and microscopic findings of premalignant and malignant lesions were noted. Special stains were done as and when required.ResultsOf the 1253 cholecystectomies received during the study period, 50 gallbladders (3.9%) showed neoplastic pathology and were included in the present study. The age range was 40 to 60 years with female predominance. Ultrasonography revealed nonspecific wall thickness in both premalignant and incidental gallbladder carcinomas. Gallstones were seen in 74% of the cases (37/50). Gross and imaging findings in 17 (34%) of the malignant cases were in concordance with microscopic features, whereas the dysplastic lesions (21) and IGBC cases showed evidence of chronic cholecystitis on the same. Microscopic examination revealed focal dysplasia (low and high grade) in 21/50 (42%) cases. Invasive malignancy was seen in 28/50 (56%) of the cases, of which 11 cases (22%) were IGBC. Pancreaticobiliary type of adenocarcinoma was the most common morphology seen in almost all the cases. There was also one case each of intracholecystic papillary neoplasm (ICPN) and carcinosarcoma.ConclusionGBC is an unusual malignancy and its preoperative diagnosis is not so definitive. The incidental form of GBC presents as a radiological disguise and a histopathological surprise. Hence, the present study warrants a complete and scrupulous histopathological examination of all the cholecystectomy specimens for proper and further management of the case.

Identification of Arhgef12 and Prkci as genetic modifiers of retinal dysplasia in the Crb1rd8 mouse model.

Mutations in the apicobasal polarity gene CRB1 lead to diverse retinal diseases, such as Leber congenital amaurosis, cone-rod dystrophy, retinitis pigmentosa (with and without Coats-like vasculopathy), foveal retinoschisis, macular dystrophy, and pigmented paravenous chorioretinal atrophy. Limited correlation between disease phenotypes and CRB1 alleles, and evidence that patients sharing the same alleles often present with different disease features, suggest that genetic modifiers contribute to clinical variation. Similarly, the retinal phenotype of mice bearing the Crb1 retinal degeneration 8 (rd8) allele varies with genetic background. Here, we initiated a sensitized chemical mutagenesis screen in B6.Cg-Crb1rd8/Pjn, a strain with a mild clinical presentation, to identify genetic modifiers that cause a more severe disease phenotype. Two models from this screen, Tvrm266 and Tvrm323, exhibited increased retinal dysplasia. Genetic mapping with high-throughput exome and candidate-gene sequencing identified causative mutations in Arhgef12 and Prkci, respectively. Epistasis analysis of both strains indicated that the increased dysplastic phenotype required homozygosity of the Crb1rd8 allele. Retinal dysplastic lesions in Tvrm266 mice were smaller and caused less photoreceptor degeneration than those in Tvrm323 mice, which developed an early, large diffuse lesion phenotype. At one month of age, Müller glia and microglia mislocalization at dysplastic lesions in both modifier strains was similar to that in B6.Cg-Crb1rd8/Pjn mice but photoreceptor cell mislocalization was more extensive. External limiting membrane disruption was comparable in Tvrm266 and B6.Cg-Crb1rd8/Pjn mice but milder in Tvrm323 mice. Immunohistological analysis of mice at postnatal day 0 indicated a normal distribution of mitotic cells in Tvrm266 and Tvrm323 mice, suggesting normal early development. Aberrant electroretinography responses were observed in both models but functional decline was significant only in Tvrm323 mice. These results identify Arhgef12 and Prkci as modifier genes that differentially shape Crb1-associated retinal disease, which may be relevant to understanding clinical variability and underlying disease mechanisms in humans.

Discriminant models of the possibility of benign nevi occurrence and features in men depending on the characteristics of anthropo-somatotypological indicators

The multifactorial nature of the origin and development of nevi is the subject of debate so far. One way to understand this process and get an answer to this question is to use a constitutional method of research. The purpose of the study is to build and analyze discriminant models of benign nevi occurrence possibility in men depending on the characteristics of the structure and size of the body. For Ukrainian men aged 22 to 35 years with benign nevi (34 with melanocyte benign simple nevi; 27 with melanocyte benign dysplastic nevi; 14 with melanocyte benign congenital nevi; 17 with nonmelanocyte benign nevus) determined anthropometric indicators according to the scheme of Bunak V. V. (1941), components of the somatotype according to the Heath-Carter scheme (1990), as well as indicators of the component composition of body weight according to Matejko formulas (1921). The control group consisted of anthropometric and somatotypological indicators of 82 practically healthy men of the same age group selected from the data bank of the Research Center of National Pirogov Memorial Medical University, Vinnytsya. Discriminant analysis was performed in the licensed statistical package “Statistica 5.5”. With the help of discriminant analysis, reliable models of the possibility of benign nevi depending on the characteristics of anthropometric and somatotypological indicators are built. It was found that healthy and patients with benign nevi of men can reliably interpret the obtained classification indicators between healthy and sick, and between patients with melanocyte simple or dysplastic nevi and other groups of benign nevi (discriminant function covers 75.7 % of cases; Wilks' Lambda statistics=0.125; р&lt;0.001). Between groups of benign nevi, reliable interpretation of the obtained classification indicators is possible only between patients with melanocyte simple or dysplastic nevi and melanocyte congenital or non-melanocyte nevi (discriminant function covers 48.4 % of cases; Wilks' Lambda statistics=0.662; р&lt;0.001), however, the totality of all anthropological variables has little discrimination. The models of healthy and sick men include the skinfold thickness (42.8 %), girth sizes (28.6 %), shoulder width and endomorphic component of the somatotype (14.3 % each); and among men with benign nevi, only girth sizes of the body. The greatest contribution to discrimination in models of healthy and sick men is made by the circumference of the forearm at the top, the width of the shoulders and the skinfold thickness on the side; and among patients with benign nevi – chest girth on inspiration. The obtained results indicate a significant influence of environmental factors on the occurrence of benign nevi.

MLH1, BRAF and p53 - searching for significant markers to predict evolution towards adenocarcinoma in colonic sessile serrated lesions.

Background and Aim: Colonic serrated lesions are premalignant lesions, using an alternative malignization pathway, including multiple genetic and epigenetic alterations, as: mismatch repair deficiency due to MutL homolog 1 (MLH1) promoter methylation, tumor protein p53 (TP53) mutations, activating mutations of v-Raf murine sarcoma viral oncogene homolog B (BRAF) and Kirsten rat sarcoma viral oncogene homolog (KRAS). Our study aims to evaluate MLH1, BRAF and p53 immunohistochemical (IHC) status in sessile serrated lesions (SSLs), with and without dysplasia. Materials and Methods: This is a retrospective case-control study including 20 SSLs with dysplasia and 20 SSLs without dysplasia (matching sex and age). IHC expression of MLH1, BRAF and p53 was evaluated as the percent of nuclear loss of MLH1, cytoplasmic positivity of BRAF and nuclear positivity of p53. Data concerning age, sex, localization of the lesion, dysplasia and IHC results were statistically processed using Microsoft Excel. Results: We had very polymorphous patterns of IHC expression for BRAF, MLH1 and p53, especially in the dysplastic group. Thus, two patients were BRAF+/MLH1-/p53+, three were BRAF+/MLH1-/p53-, one was BRAF+/MLH1+/p53- and six were BRAF+/MLH1+/p53+. Dysplastic lesions without BRAF mutation exhibited the following phenotype: one case BRAF-/MLH1-/p53+, four BRAF-/MLH1-/p53- and three BRAF-/MLH1+/p53+. In the control group (SSLs without dysplasia), there was a more homogenous distribution of cases: eight cases BRAF+/MLH1+/p53-, seven BRAF-/MLH1+/p53-, one BRAF-/MLH1-/p53+, two BRAF-/MLH1-/p53- and two BRAF-/MLH1+/p53+. Conclusions: There are more routes on the serrated pathway, with different mutations and time of acquisition of each genetic or epigenetic lesion with the same morphological result. These lesions should be stratified according to their risk to poor outcome and their need to further surveillance.

Oral squamous cell carcinomas: a histopathological review of multiple cases from Western Romania.

Malignant tumors of the oral cavity have a growing incidence, most being squamous cell carcinomas, generally called oral cancers (OCs), clinically detected at various stages of natural evolution. The increased incidence in Romania in recent years and the lack of conclusive data have led to the development of this study. The main purpose of this study was to assess the molecular profile of tumors, the types of blood vessels associated with the tumor, and expression of tumor immunomarkers. Regarding morphological findings, focal epithelial hyperplasia, dysplastic lesions, typical mitoses, perineural invasion, parakeratosis and keratosis beads, intracytoplasmic keratinization were observed. Microvascular density was higher in the tumor area compared to the peritumoral area. Lymphovascular invasion was identified in 13% of cases, which also presented regional lymph node metastases. Podoplanin expression was identified in 79% of cases which were tested positive for the D2-40 immunomarker. All p53-positive cases co-expressed epidermal growth factor receptor (EGFR), half of the EGFR-positive cases co-expressed p53, and co-expression of CD117 and p63 was identified in 80% of EGFR-positive/cytokeratin 5 (CK5)-positive cases being proposed the basal-like subtype of OCs, defined as EGFR-positive/CK5-positive, CD117-positive and p63-positive. Results support the need for molecular classification of OCs based on of tumor immunomarker expression and gene analysis.

Hide and Seek: Dysplasia Camouflage in a Patient With Barrett’s Esophagus

Question: An 84-year-old man presenting with fatigue was referred for endoscopy owing to anemia. On his esophagogastroduodenoscopy, a segment of Barrett’s esophagus (BE) was seen (Prague classification C2M4). Random biopsies were obtained as per the Seattle protocol. Histology revealed at least one specimen harboring high-grade dysplasia. He had ascending colon adenocarcinoma on colonoscopy. The BE segment was subsequently reassessed using white light, narrow band imaging (NBI), acetic acid chromoendoscopy, and high magnification. In a proximal tongue, there was an area appeared to have an irregular microvasculature. This was presumed to be the dysplastic lesion (Figure A; NBI). A clear demarcation line was noted between the distal nondysplastic segment and the irregular microvasculature of the proximal dysplastic area (Figure B; NBI and near focus). A decision was made to initially resect the BE segment using the endoscopic submucosal dissection (ESD) technique and operate the right-sided colon cancer at the same season in the following weeks. He continued his proton pump inhibitor (PPI) in the interim. While reassessing the lesion before performing ESD 0 month later, the previously identified dysplastic lesion was no longer identified (Figure C; white light). What is the most likely explanation and what is the best way to manage the case? See the Gastroenterology web site () for more information on submitting your favorite image to Clinical Challenges and Images in GI. Further careful examination with NBI and magnification revealed an area of brownish discoloration below re-epithelialized mucosa (Figure D). The area to resect could not be ascertained, but the NBI and magnification images combined with the previously known location of the dysplastic area were used as landmarks to proceed with an ESD (Figure E, F). On histology, the dysplastic BE was buried underneath normal appearing squamous cell epithelium (Figure G; stain: hematoxylin and eosin). The dysplastic glandular proliferation showed focally fused glands formed by atypical, mitotically active epithelial cells without goblet cells. The dysplastic elements did not involve the muscularis mucosae nor the submucosa. This unique case demonstrates that re-epithelialization of a Barrett’s mucosa can occur within a short time interval. While managing BE, endoscopists should be aware of the acceleration of re-epithelization and the potential concealing of dysplastic cells under normal-appearing epithelial tissue such as presented here. Some evidence has shown that acid suppression can lead to this occurrence. A study of 26 patients examining the effect of high-dose lansoprazole on BE found that squamous islands were developed in 77% of the patients by the final visit.1Sampliner R.E. Effect of up to 3 years of high-dose lansoprazole on Barrett’s esophagus.Am J Gastroenterol. 1994; 89: 1844-1848PubMed Google Scholar In the study, squamous re-epithelization and subsequent burying of the dysplasia below the surface could be attributed to high-dose PPI use. It is, thus, paramount to carefully inspect the whole BE segment and beyond its obvious extent, especially in patients on a PPI. Likewise, this observation highlights the utmost importance of careful inspection and precise documentation of the location of dysplasia in BE during endoscopy. Applying advanced imaging techniques in addition to the white light may facilitate the visibility and improve the diagnostic and interventional precision.2Qumseya B. Sultan S. Bain P. et al.ASGE guideline on screening and surveillance of Barrett’s esophagus.GIE Journal. 2019; : 335-359Google Scholar

Evaluation of the Distribution of Candida Species in Patients with Dysplastic and Nondysplastic Oral Lichen Planus Lesions.

Objectives This study is aimed at identifying and determining the distribution of isolated Candida species in patients with dysplastic and nondysplastic oral lichen planus (OLP) lesions in comparison with those of healthy controls. Material and Methods. This study includes patients with OLP, aged (more than 18 years old), who have had informed consent. Samples of the oral, tongue, and buccal mucus by rubbing with a sterile swab and sterilely next to the lamp flame. Demographic information was obtained using patient records to determine the species of Candida in both groups, and two tests of fertile tube production by Candida albicans and dye production in the dye medium were used. A biopsy from OLP lesions has been taken from each patient after swab sampling and was sent to the pathology department for further histopathological analysis. In the end p value, less than 0/05 was considered significant. Result In this study, 40 lichen planus patients were compared with 32 control patients. The female/male ratio in OLP and healthy groups was 22/18 and 17/15, respectively. Among the OLP patients, 23 cases (56%) were dysplastic, and the other 17 (44%) patients were nondysplastic. The mean (±standard deviation (SD)) age of patients was 48.83 (±9.34) years, and the mean age of the control group was 40.21 (±10.32). There were no significant differences based on age (p > 0.05). The highest frequency was related to tongue in both groups (22 (55%)) and buccal mucosa was the least common. There was a significant relationship between the location of the lesion and OLP (p = 0.05). 18 (45%) were erosive, and 22 (55%) were nonerosive. However, no significant difference was observed between erosive and nonerosive types in the OLP group (p = 0/07). Regarding the type of Candida, all cases in the patient's group were related to Candida albicans [40 (100%)], and the correlation was not found in this regard (p > 0/05). About colony count, the mean for the case and control groups was 26.68 and 23.25, respectively. Also, no significant relationship was found between colony count and groups in this study (p = 0.3). There was no significant difference between gender and dysplastic or nondysplastic (p > 0.05). Conclusion According to the statistical studies performed in this study, the presence of Candida in patients with dysplastic and nondysplastic lichen planus is not significantly different, and this rate is not higher than healthy individuals and in cases where the results are positive. The predominant species of Candida is the Candida albicans. In this study, the highest frequency was related to tongue in both groups. There was only a significant relationship between the location of the lesion and OLP.

Multicenter prospective clinical trial of molecular genetic markers for non-invasive differential diagnosis of benign and malignant melanocytic skin lesions.

e21581 Background: Improving the accuracy of non-invasive diagnosis of skin tumors is becoming an urgent problem, given the clear increase in the incidence of melanoma in many countries. Dermoscopy and examination by an experienced dermatologist can reduce the NNE to 5–10 per melanoma (M) detected, however, achieving even this, clearly suboptimal, rate requires a long specialist training. Methods: We report results of prospective nonrandomized trial evaluating sensitivity and specificity of RNA profiling of cytological samples from adhesive transparent patches (ATP) applied to skin lesions. Routine pathology reports were obtained for all surgery samples. RNA extracted from ATP specimens (N = 126) was tested for mRNA expression of PRAME and LINC00518 genes. GAPDH was used as a reference gene. Sample was considered marker-positive if either PRAME or LINC00518 expression was detected. Results: Between June 2021 and November 2021, 126 pts undegoing excisional biopsy of skin lesions were included in the study at 5 centers. On pathology, invasive M was detected in 49 (38.9%) patients (mean Breslow thickness was 2.41 mm [95%CI 1,44 to 3.37]), M in situ in 6 (4.8%), non-melanoma skin cancer in 4 (3.2%) and dysplastic nevus in 56 (44.4%). In the day of preplanned excision ATP was applied to the target lesion for 5–6 min and sent to the central lab. Amplifiable mRNA was found in all 126 ATP samples. 82 samples of 126 (65,1%) tested marker-positive. Sensitivity of RNA profiling for detection of skin cancers reached 88.14% (95% CI 77.07% – 95.09%), while specificity was only 53.85% (41.03% – 66.30%), with PPV of 63.41% (56.74% – 69.61%), NPV of 83.33% (70.65% – 91.22%) and accuracy of 70.16% (61.29% – 78.04%). Among the seven false negative results there were 3 M in situ and 4 invasive M. Conclusions: RNA profiling of skin ATP specimens is feasible and has high sensitivity but rather low specificity. Randomized studies are needed to evaluate if this technique can add anything to dermoscopy or other noninvasive diagnostic modalities. Clinical trial information: NCT04353050. [Table: see text]