Decoding the Rarity: A Unique Case of Clear-Cell Variant of Oral Squamous Cell Carcinoma.

The clear-cell variant of oral squamous cell carcinoma is an extremely rare histological variant and an incompletely understood entity. Clear cell appearance in squamous cell carcinoma may be attributed to hydropic degeneration of neoplastic cells. We report a case of a 32-year-old male patient who presented with an ulceroproliferative growth in the left maxillary posterior region on the hard palate and gingiva, obliterating the buccal vestibule. Histopathologic examination revealed thick anastomosing strands of round to ovoid neoplastic cells with predominantly clear cytoplasm and marked cellular and nuclear pleomorphism infiltrating into the fibro-cellular connective tissue stroma. Special staining and immunohistochemistry (IHC) were performed to rule out the differentials of clear-cell variants of different sites such as salivary gland, odontogenic origin, and metastatic tumors. The clear cells were negative for periodic acid Schiff (PAS) and mucicarmine. The malignant clear cells showed positive reactions with IHC markers pan-cytokeratin and P63 and yielded negative results for S100 and CD10, confirming the diagnosis as a clear-cell variant of oral squamous cell carcinoma. We emphasize the importance of prompt and comprehensive diagnostic work-up to identify this rare, aggressive, and possibly fatal neoplasm.

Myxomatous Mysteries: Decoding the Oral Focal Mucinosis.

Oral focal mucinosis (OFM) is a unique benign lesion of the oral cavity with uncertain etiology which is analogous to cutaneous focal mucinosis. It mainly affects women in their fourth and fifth decades of life. The diagnosis of this condition is based on histopathological examination, as it lacks characteristic clinical and radiographic features. Its pathophysiology is associated with fibroblasts producing excessive amounts of hyaluronic acid, which causes localized myxomatous changes. Here, we describe the occurrence of this rare entity in a 54-year-old female patient involving attached gingiva of the left posterior mandibular region along with emphasis on its histopathological and histochemical findings to differentiate it from clinically and microscopically look-alike lesions.

Chemical speciation and health risk assessment of potentially toxic elements in playground soil of bell metal commercial town of Eastern India.

Contaminated playground soils can expose players to harmful pollutants, increasing the risk of respiratory, skin, and gastrointestinal issues and potentially impacting long-term health and development. This study investigated the chemical forms and the human health risks associated with potentially toxic elements (PTEs) found in playground soil samples from Khagra, a historic town known for its bell metal industry, located in the Murshidabad district of eastern India. Sequential extraction techniques were employed to analyze the distribution of PTEs such as As, Cd, Co, Cu, Mn, Pb, Ni, Sn, and Zn among different fractions: exchangeable (F1), bound to carbonate phase (F2), bound to iron and manganese oxides (F3), bound to organic matter (F4), and residual (F5). The playground soil showed the highest contamination with Sn, with an IPOLL value of 3.14, indicating moderate to heavy contamination, while Cd, Cu, Mn, Pb, and Zn exhibit moderate contamination. The mean concentration of PTEs in all fractions (F1-F5) follows the order: Fe > Zn > Cu > Mn > Pb > Sn > Ni > Co > As > Cd. The maximum affinity of PTEs and their percentages are as follows: Fe (F5, 80.6%), As (F5, 55.31%), Cd (F5, 48.8), Co (F5, 64.9%), Mn (F3, 44%), Ni (F5, 53.2%), Pb (F3, 44.7%), Zn (F3, -43.19%), Sn (F3, 55%), Cu (F5 -42.18). As, Cd, Co, Cu, Fe, and Ni have a high affinity for F5, indicating geogenic source, while Mn, Pb, Sn, and Zn have a high affinity for F3, indicating anthropogenic source. Fe-Mn oxide partition was dominant for nearly all PTEs due to elevated sorption of cations onto Fe-Mn oxides at high pH. The risk assessment code for Cd, Cu, Mn, Ni, Sn, and Zn in playground soil is categorized under moderate risk, below 30%, while other elements showed no risk. Also, mobility factors were calculated for each PTEs, suggesting the degree of mobility that PTEs can easily migrate and be taken up, absorbed, or adsorbed by the human body. The mobility factor in playground soil was higher for Sn (59.89%) followed by Mn (54.24%) > Pb (52.91%) > Zn (52.01%) > Cd (39.49%) > Ni (33.20%) > As (30.39%) > Co (26.56%) > Cu (21.24%) > Fe (11.20%). Risk hazard quotients for children and adults were found to follow the order: Pb (0.263; 0.040), Cu (0.098; 0.015) > As(0.056; 0.008) > Mn (0.045; 0.009) > Zn(0.36; 0.05) > Cd(0.006; 0.001) > Ni (0.004; 0.001) > Co (0.001; 0.0). PTEs detected in the environment result from atmospheric deposition from small-scale metallurgical industries (bell metal and brass), coal and oil combustion, civil works, municipal waste incineration, and fugitive emissions from road dust. The human non-carcinogenic health risk for PTEs from ingestion and dermal contact was higher than that from inhalation. In the context of carcinogenic risk, As shows the highest health risk of 2.51E-05, followed by Cd (1.02E-09) and Co (8.14E-09). This study uniquely assesses the chemical speciation of PTEs in playground soils, revealing their geogenic and anthropogenic sources, and evaluates associated health risks. Policy intervention is vital for monitoring and remediating PTEs in playgrounds to protect children's health.

Oral Granulomatous Disorders: A Diagnostic Insight.

Granulomatous inflammation represents a unique pattern of chronic inflammation observed in a restricted form of infectious and certain non-infectious diseases. The formation of granulomas typically involves immune responses. Granulomatous disorders encompass a broad spectrum of conditions that share the common histological feature of granuloma formation. Their involvement in the oral soft and hard tissues is quite infrequent; however, their manifestation can pose a diagnostic challenge due to the diverse range of potential causes and the relatively non-specific appearance of the individual lesions. The ultimate outcome of a complex entails the formation of a granuloma, resulting from the interplay among an invading pathogen or antigen, chemical substance, medication, or other irritant, persistent presence of antigens in the bloodstream, activation of macrophages, initiation of Th1 cell response, B-cell overactivity, presence of circulating immune complexes, and a wide range of biological signaling molecules, ultimately leading to the development of fibrosis attributed to the actions of transforming and platelet-derived growth factor. This article emphasizes the clinicopathological diagnostic criteria of oral granulomatous disorders as a guide for treatment and management.

Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy.

Extracellular matrix diseases like fibrosis are elusive to diagnose early on, to avoid complete loss of organ function or even cancer progression, making early diagnosis crucial. Imaging the matrix densities of proteins like collagen in fixed tissue sections with suitable stains and labels is a standard for diagnosis and staging. However, fine changes in matrix density are difficult to realize by conventional histological staining and microscopy as the matrix fibrils are finer than the resolving capacity of these microscopes. The dyes further blur the outline of the matrix and add a background that bottlenecks high-precision early diagnosis of matrix diseases. Here we demonstrate the multiple signal classification method-MUSICAL-otherwise a computational super-resolution microscopy technique to precisely estimate matrix density in fixed tissue sections using fibril autofluorescence with image stacks acquired on a conventional epifluorescence microscope. We validated the diagnostic and staging performance of the method in extracted collagen fibrils, mouse skin during repair, and pre-cancers in human oral mucosa. The method enables early high-precision label-free diagnosis of matrix-associated fibrotic diseases without needing additional infrastructure or rigorous clinical training.

Fibroblastic osteosarcoma: A perplexing entity.

Osteosarcoma (OS), also referred to as osteogenic sarcoma, is the most common primary malignant tumour affecting long bones, characterised by the proliferation of osteoblastic precursor cells and the production of osteoid or immature bone. OSs of the head and neck region have unique biology, exhibiting a clinical behaviour and a natural history that are distinct from OSs of the trunk and extremities. Similarly, their radiological appearance and histological growth pattern can be quite diverse proving to be a challenge to histopathologists to arrive at a diagnosis. Hence, OSs of the jaw remain enigmatic, and a number of difficulties related to their diagnosis and treatment are yet to be resolved. This article reports on a case of advanced OS of the mandible in a 45-year-old woman who came for the evaluation of swelling. This case illustrates the various modalities of diagnosis, such as radiology, histopathology and immunohistochemistry for the confirmation of the variants of OS, leading to an enormously improved quality of life by offering an improved opportunity for cure and treatment.

Portable, handheld, and affordable blood perfusion imager for screening of subsurface cancer in resource-limited settings.

Precise information on localized variations in blood circulation holds the key for noninvasive diagnostics and therapeutic assessment of various forms of cancer. While thermal imaging by itself may provide significant insights on the combined implications of the relevant physiological parameters, viz. local blood perfusion and metabolic balance due to active tumors as well as the ambient conditions, knowledge of the tissue surface temperature alone may be somewhat inadequate in distinguishing between some ambiguous manifestations of precancer and cancerous lesions, resulting in compromise of the selectivity in detection. This, along with the lack of availability of a user-friendly and inexpensive portable device for thermal-image acquisition, blood perfusion mapping, and data integration acts as a deterrent against the emergence of an inexpensive, contact-free, and accurate in situ screening and diagnostic approach for cancer detection and management. Circumventing these constraints, here we report a portable noninvasive blood perfusion imager augmented with machine learning-based quantitative analytics for screening precancerous and cancerous traits in oral lesions, by probing the localized alterations in microcirculation. With a proven overall sensitivity >96.66% and specificity of 100% as compared to gold-standard biopsy-based tests, the method successfully classified oral cancer and precancer in a resource-limited clinical setting in a double-blinded patient trial and exhibited favorable predictive capabilities considering other complementary modes of medical image analysis as well. The method holds further potential to achieve contrast-free, accurate, and low-cost diagnosis of abnormal microvascular physiology and other clinically vulnerable conditions, when interpreted along with complementary clinically evidenced decision-making perspectives.

Collagen deposition correlates with loss of E-cadherin and increased p63 expression in dysplastic conditions of oral submucous fibrosis.

This paper focuses on the status of epithelial markers, E-cadherin, and p63 in the backdrop of an abnormal amount of collagen in the sub-mucosa of dysplastic and non-dysplastic grades of OSF. Histologically confirmed OSF and normal oral mucosa samples were procured. Samples were stained by Van Gieson's stain (VG) and immunohistochemistry. The captured images were analyzed by ImageJ software to quantify their grayscale intensities. There was a gradual increase in the intensity of VG stain from normal to non-dysplastic and dysplastic OSF and the differences in their mean grayscale values were found to be significant (p < 0.00001). The intensity of E-cadherin was found to be the highest in non-dysplastic conditions and lowest in dysplastic conditions. The intensity difference of E-cadherin between normal and non-dysplastic OSF was found to be significant (p < 0.00001). The grayscale scale intensity values for p63 in whole epithelium depicted significant differences between normal and diseased conditions but for its intensity, in basal cells, significant differences were found between non-dysplastic and other classes of tissues. There was a positive correlation observed between VG and p63 staining intensity. The diseased oral epithelium demonstrated greater deposition of sub-epithelial collagen fibers along with subsequent loss of E-cadherin and an increased p63 expression.

Comprehensive Evaluation of PAXgene Fixation on Oral Cancer Tissues Using Routine Histology, Immunohistochemistry, and FTIR Microspectroscopy.

The choice of tissue fixation is critical for preserving the morphology and biochemical information of tissues. Fragile oral tissues with lower tensile strength are challenging to process for histological applications as they are prone to processing damage, such as tissue tear, wrinkling, and tissue fall-off from slides. This leads to loss of morphological information and unnecessary delay in experimentation. In this study, we have characterized the new PAXgene tissue fixation system on oral buccal mucosal tissue of cancerous and normal pathology for routine histological and immunohistochemical applications. We aimed to minimize the processing damage of tissues and improve the quality of histological experiments. We also examined the preservation of biomolecules by PAXgene fixation using FTIR microspectroscopy. Our results demonstrate that the PAXgene-fixed tissues showed significantly less tissue fall-off from slides. Hematoxylin and Eosin staining showed comparable morphology between formalin-fixed and PAXgene-fixed tissues. Good quality and slightly superior immunostaining for cancer-associated proteins p53 and CK5/6 were observed in PAXgene-fixed tissues without antigen retrieval than formalin-fixed tissues. Further, FTIR measurements revealed superior preservation of glycogen, fatty acids, and amide III protein secondary structures in PAXgene-fixed tissues. Overall, we present the first comprehensive evaluation of the PAXgene tissue fixation system in oral tissues. This study concludes that the PAXgene tissue fixation system can be applied to oral tissues to perform diagnostic molecular pathology experiments without compromising the quality of the morphology or biochemistry of biomolecules.

Pathophysiological relationship between hypoxia associated oxidative stress, Epithelial-mesenchymal transition, stemness acquisition and alteration of Shh/ Gli-1 axis during oral sub-mucous fibrosis and oral squamous cell carcinoma.

Oral sub-mucous fibrosis (OSF) is a pathophysiological state of oral cavity or oropharynx having a high chance of conversion to oral squamous cell carcinoma (OSCC). It involves fibrotic transformation of sub-epithelial matrix along with epithelial abnormalities. The present work aims to unveil the mechanistic domain regarding OSF to OSCC conversion exploring the scenario of hypoxia associated oxidative stress, epithelial-mesenchymal transition (EMT), metastasis and stemness acquisition. The study involves histopathological analysis of the diseased condition along with the exploration of oxidative stress status, assessment of mitochondrial condition, immunohistochemical analysis of HIF-1α, E-cadherin, vimentin, ERK, ALDH-1, CD133, Shh, Gli-1 and survivin expressions in the oral epithelial region together with the quantitative approach towards collagen deposition in the sub-epithelial matrix. Oxidative stress was found to be associated with type-II EMT in case of OSF attributing the development of sub-epithelial fibrosis and type-III EMT in case of OSCC favoring malignancy associated metastasis. Moreover, the acquisition of stemness during OSCC can also be correlated with EMT. Alteration of Shh and Gli-1 expression pattern revealed the mechanistic association of hypoxia with the phenotypic plasticity and disease manifestation in case of OSF as well as OSCC. Shh/ Gli-1 signaling can also be correlated with survivin mediated cytoprotective phenomenon under oxidative stress. Overall, the study established the correlative network of hypoxia associated oxidative stress, EMT and manifestation of oral pre-cancerous and cancerous condition in a holistic approach that may throw rays of hope in the therapeutic domain of the concerned diseases.

Multifractal Alterations in Oral Sub-Epithelial Connective Tissue During Progression of Pre-Cancer and Cancer.

Bright-field microscopy (BFM) encrypts the optical transillumination profile of the transmitted light attenuated by the complex micro-structural tissue convolutions, manifested by the dense and compact regions of the specimen under examination. The connotations of idiosyncratic tissue interaction dynamics with the onset of pre-cancerous activity are encoded in the BFM acquired oral mucosa histopathological images (OMHI). In the present study, our analysis is focused on the sub-epithelium region of the oral mucosa, which has high clinical significance but sparsely explored in the literature from the textural domain. Histopathology being the gold-standard technique till date, we have used the light microscopic histopathology images for tissue characterization. The tissue-index transmission patches (TITP) from the sub-epithelium region are cropped under the guidance of oral onco-pathologists. After that, the TITPs are characterized for its multi-scale spatial-deformation dynamics, while keeping the intrinsic anisotropic geometry, and local contour connectivity within tolerable limits. With recent studies exhibiting multifractal's potency in diverse biological system analysis, here, we exploit the 2D multifractal detrended fluctuation analysis (2D-MFDFA) on TITPs for exploring a discriminative set of multifractal signatures for healthy, oral potentially malignant disorders and oral cancer tissue sample. The predictive model's competency is validated on an experimentally collected corpus of TITP samples and substantiated via confirmatory data statistics and analysis, showing its inter-class segregation efficacy. Moreover, the 2D-MFDFA analysis evinces the complex multifractal patterns in TITPs, which is due to the presence of composite long-range correlations in the oral mucosa tissue fabric.

Epithelial Distribution of E-Cadherin, p63, and Mitotic Figures in ApoTome Images to Determine the Oncogenic Potentiality of Oral Submucous Fibrosis.

The exact process of the malignant conversion of oral submucous fibrosis (OSF) to oral cancer is not fully understood. This study aimed to detect and analyze E-cadherin expression, p63 expression, and number of mitotic figures, all correlated to cancer development, in ApoTome images of oral tissues to determine the oncogenic potentiality of OSF. ApoTome images of the study groups (6 normal, 16 OSF with dysplasia, and 10 OSF without dysplasia) were recorded. Cytoplasmic and membranous E-cadherin expression, breakages of the cell membrane, and p63 expression were detected in MATLAB 2016b. The number of mitotic figures detected by MATLAB was correlated with the number of chromosomes detected by ImageJ. A Mann­Whitney U test was done to determine a significant difference between the study groups for cytoplasmic and membranous E-cadherin distribution points. Statistical significant differences were found for cytoplasmic E-cadherin distribution between normal and OSF (with dysplasia) (p = 0.0278). There was an increase in mitotic figures, p63 expression, and cytoplasmic E-cadherin expression and a decrease in membranous E-cadherin expression from normal to diseased condition. Hence, automated detection and quantification of E-cadherin, p63, and mitotic figures in ApoTome images of oral biopsies can help in determining the oncogenic potentiality of OSF.

Malignant potentiality assessment of oral submucous fibrosis through semi-quantitative approach.

BACKGROUND: In the context of early diagnosis and prevention of oral cancer, precise assessment of malignant potentiality of the oral potentially malignant disorders, particularly oral submucous fibrosis (OSF) is crucial. Till date, the assessment of malignant potentiality suffers from predictive ambiguity due to the lack of precision in the gold standard techniques. This can be addressed by integrating heuristic domain knowledge with quantitative analysis. AIM: The aim of this study is to propose an index for enhancing accuracy in malignant potentiality evaluation. MATERIALS AND METHODS: The present study analyzes important histomorphometric attributes (epithelial thickness, basal cell nuclear size, nuclear-to-cytoplasmic area ratio of basal cells, chromaticity of basal cell nucleus, thickness of basement membrane, ratio of vasculature in juxta-epithelial connective tissue [i.e., area covered by blood vessels/total area], collagen density in the lamina propria) of oral mucosa in dysplastic and nondysplastic OSF in association with relevant oncopathological appreciations (weightage of different features as suggested by oral pathologists) toward proposing a "Malignant Potentiality Index" (MPI). RESULTS: Analysis of variance and notch box plot analysis depict statistically significant differences (P < 0.0001) in the histopathological features among different study groups (normal oral mucosa, OSF without dysplasia, OSF with dysplasia). Histopathological observation of one OSF patient with calculated MPI is shown. CONCLUSION: This newly proposed diagnostic cum prognostic decision-making parameter, the "MPI" may bring a value addition to the conventional diagnostic gold standard.

A miscellany of cribriform pattern, squamous metaplasia and clear cells in pleomorphic adenoma of upper lip: A diagnostic paradox.

Pleomorphic Adenoma (PA), a benign neoplasm of glandular origin most commonly involves major salivary glands. It is rare in minor salivary glands such as hard palate, upper lip and buccal mucosa, frequently affecting middle aged females. PA comprises diverse histopathologic features of epithelial, myoepithelial and mesenchymal components. Aberrant histopathologic features in Pleomorhic Adenoma thus calls for judicious discrimination from alike entities which facilitates appropriate surgical management. Here we present a case report of PA in upper lip in a 25 year old female patient showing uncommon findings like clear cells, squamous metaplasia and cribriform pattern.

Rhabdomyosarcoma involving maxilla mimicking gingival enlargement: A diagnostic challenge.

Rhabdomyosarcoma (RMS) is a rare, rapidly growing, highly aggressive malignant neoplasm, originating from undifferentiated mesenchymal cells that retain their ability to differentiate into skeletal muscle. It mainly affects children, accounts for <1% of all adult malignancies and has varied clinical presentations. The head and neck region accounts for 35%-40% of all RMS cases, of which 10%-12% cases involve the oral cavity. This report deals with a case of RMS in a 40-year-old woman, primarily involving maxillary gingiva for which she underwent excision with subsequent recurrences. The uniqueness of this case is that it reminds us of the essential clinical dictum that 'every growth we encounter, no matter how benign it appears clinically, should be looked upon with suspicion'. Hence, proper integration of history, clinical examination and investigation is required to reach a correct diagnosis enabling early treatment, thereby preventing functional and aesthetic loss and psychological trauma.

Elucidation of Differential Nano-Textural Attributes for Normal Oral Mucosa and Pre-Cancer.

Computational analysis on altered micro-nano-textural attributes of the oral mucosa may provide precise diagnostic information about oral potentially malignant disorders (OPMDs) instead of an existing handful of qualitative reports. This study evaluated micro-nano-textural features of oral epithelium from scanning electron microscopic (SEM) images and the sub-epithelial connective tissue from light microscopic (LM) and atomic force microscopic (AFM) images for normal and OPMD (namely oral sub-mucous fibrosis, i.e., OSF). Objective textural descriptors, namely discrete wavelet transform, gray-level co-occurrence matrix (GLCM), and local binary pattern (LBP), were extracted and fed to standard classifiers. Best classification accuracy of 87.28 and 93.21%; sensitivity of 93 and 96%; specificity of 80 and 91% were achieved, respectively, for SEM and AFM. In the study groups, SEM analysis showed a significant (p < 0.01) variation for all the considered textural descriptors, while for AFM, a remarkable alteration (p < 0.01) was only found in GLCM and LBP. Interestingly, sub-epithelial collagen nanoscale and microscale textural information from AFM and LM images, respectively, were complementary, namely microlevel contrast was more in normal (0.251) than OSF (0.193), while nanolevel contrast was more in OSF (0.283) than normal (0.204). This work, thus, illustrated differential micro-nano-textural attributes for oral epithelium and sub-epithelium to distinguish OPMD precisely and may be contributory in early cancer diagnostics.

Segmentation and analysis of surface characteristics of oral tissues obtained by scanning electron microscopy to differentiate normal and oral precancerous condition.

Abnormal epithelial stratification is a sign of oral dysplasia and hence evaluation of surface characteristics of oral epithelial region can help in detection of cancerous progression. Surface characteristics can be better visualised by Scanning Electron Microscopy (SEM) in comparison to light microscopy. In our study we have developed automated image processing algorithms i.e. Gaussian with median filtering and Gradient filtering, using MATLAB 2016b, to segment the surface characteristics i.e. the ridges and pits in the SEM images of oral tissue of normal (13 samples) and Oral Submucous Fibrosis (OSF) (36 samples) subjects. After segmentation, quantitative measurement of the parameters like area, thickness and textural features like entropy, contrast and range filter of ridges as well as area of pit and the ratio of area of ridge vs. area of pit was done. Statistical significant differences were obtained in between normal and OSF study groups for thickness (p=0.0107), entropy (p<0.00001) and contrast of ridge (p<0.00001) for Gaussian with median filtering and for all the parameters except thickness of the ridge(p=1.386), for Gradient filtering. Thus, computer aided image processing by Gradient filter followed by quantitative measurement of the surface characteristics provided precise differentiation between normal and precancerous oral condition.

Genome-wide miRNA methylome analysis in oral cancer: possible biomarkers associated with patient survival.

AIM: The methylome associated with miRNA loci was investigated in oral cancer to explore tobacco specific methylation and potential biomarkers for patient survival. METHODS: Methylome data was generated from 16 pairs of cancer-normal tissues by reduced representation bisulfite sequencing method. Differentially methylated regions were identified using the DMAP pipeline. In silico validation and Kaplan-Meier survival analyses were performed on The Cancer Genome Atlas data based on our miRNA methylome data. RESULTS: A total of 4310 unique differentially methylated regions, mapping to 144 miRNA loci, were identified. Three distinct groups of miRNAs were differentially methylated in cancer tissues from smokers, chewers and mixed habitués. Hypermethylation of miR-503, miR-200a/b, miR-320b and miR-489 was associated with worse 5-year survival. CONCLUSION: Differential methylation patterns in miRNA loci are associated with poor survival underscoring their potential as predictive and prognostic biomarkers in oral cancer.