Development of Automated Risk Stratification for Sporadic Odontogenic Keratocyst Whole Slide Images with an Attention-Based Image Sequence Analyzer.

(1) Background: The categorization of recurrent and non-recurrent odontogenic keratocyst is complex and challenging for both clinicians and pathologists. What sets this cyst apart is its aggressive nature and high likelihood of recurrence. Despite identifying various predictive clinical/radiological/histopathological parameters, clinicians still face difficulties in therapeutic management due to its inherent aggressive nature. This research aims to build a pipeline system that accurately detects recurring and non-recurring OKC. (2) Objective: To automate the risk stratification of OKCs as recurring or non-recurring based on whole slide images (WSIs) using an attention-based image sequence analyzer (ABISA). (3) Materials and methods: The presented architecture combines transformer-based self-attention mechanisms with sequential modeling using LSTM (long short-term memory) to predict the class label. This architecture leverages self-attention to capture spatial dependencies in image patches and LSTM to capture sequential dependencies across patches or frames, making it suitable for this image analysis. These two powerful combinations were integrated and applied on a custom dataset of 48 labeled WSIs (508 tiled images) generated from the highest zoom level WSI. (4) Results: The proposed ABISA algorithm attained 0.98, 1.0, and 0.98 testing accuracy, recall, and area under the curve, respectively, whereas VGG16, VGG19, and Inception V3, standard vision transformer attained testing accuracies of 0.80, 0.73, 0.82, 0.91, respectively. ABISA used 58% fewer trainable parameters than the standard vision transformer. (5) Conclusions: The proposed novel ABISA algorithm was integrated into a risk stratification pipeline to automate the detection of recurring OKC significantly faster, thus allowing the pathologist to define risk stratification faster.

Building Automation Pipeline for Diagnostic Classification of Sporadic Odontogenic Keratocysts and Non-Keratocysts Using Whole-Slide Images.

The microscopic diagnostic differentiation of odontogenic cysts from other cysts is intricate and may cause perplexity for both clinicians and pathologists. Of particular interest is the odontogenic keratocyst (OKC), a developmental cyst with unique histopathological and clinical characteristics. Nevertheless, what distinguishes this cyst is its aggressive nature and high tendency for recurrence. Clinicians encounter challenges in dealing with this frequently encountered jaw lesion, as there is no consensus on surgical treatment. Therefore, the accurate and early diagnosis of such cysts will benefit clinicians in terms of treatment management and spare subjects from the mental agony of suffering from aggressive OKCs, which impact their quality of life. The objective of this research is to develop an automated OKC diagnostic system that can function as a decision support tool for pathologists, whether they are working locally or remotely. This system will provide them with additional data and insights to enhance their decision-making abilities. This research aims to provide an automation pipeline to classify whole-slide images of OKCs and non-keratocysts (non-KCs: dentigerous and radicular cysts). OKC diagnosis and prognosis using the histopathological analysis of tissues using whole-slide images (WSIs) with a deep-learning approach is an emerging research area. WSIs have the unique advantage of magnifying tissues with high resolution without losing information. The contribution of this research is a novel, deep-learning-based, and efficient algorithm that reduces the trainable parameters and, in turn, the memory footprint. This is achieved using principal component analysis (PCA) and the ReliefF feature selection algorithm (ReliefF) in a convolutional neural network (CNN) named P-C-ReliefF. The proposed model reduces the trainable parameters compared to standard CNN, achieving 97% classification accuracy.

Inverse correlation of CD3 & vascular endothelial growth factor in incisional oral squamous cell carcinoma biopsies predicts nodal metastasis & poor survival of patients.

Background & objectives: Oral squamous cell carcinoma (OSCC) is widely prevalent in the Indian subcontinent mainly due to habit-associated aetiologies. Immune regulation and angiogenesis are the part of tumourigenesis that play a crucial role in metastasis and survival. However, the concurrent expression of vascular endothelial growth factor (VEGF) and CD3 (immune regulator receptor on T-lymphocyte) in the same OSCC tissue samples has not been reported in the Indian population. The present study evaluated the expression of CD3+ T-cells and VEGF in OSCC tissue samples and studied the clinicopathological correlation and survival analysis in an Indian population. Methods: This was a retrospective study conducted on 30 formalin-fixed and paraffin embedded sections which were histologically diagnosed as OSCC cases comprising of 15 metastatic OSCC and 15 non-metastatic OSCC with available clinical data and survival status. Results: Reduced expression of CD3+ T-cells and increased VEGF expression were observed in metastatic OSCC samples. The correlation of expression of CD3+ T-cells and VEGF with clinicopathological parameters showed a significant association between these markers with age, nodal status, site of the lesion and survival. Interpretation & conclusions: Reduced expression of CD3+ T-cells in OSCC was found to be associated with a significantly poor survival. VEGF was found to be over expressed in metastatic OSCC as compared to that in non-metastatic OSCC. The study findings suggest that the evaluation of CD3 and VEGF in incisional OSCC biopsies can be considered for predicting the survival outcome and metastasis.

Expansion microscopy: A revolution in diagnostic pathology.

Expansion microscopy (ExM) is a recent discovery in diagnostic pathology and microbiology that promotes a physical magnification of the tissue specimen instead of optical magnification. It not only improves the resolution of the specimen but also enhances the diagnostic precision, and permits nanoscale imaging of the specimen. Optical microscopy is routinely used in histopathology and microbiology for ages. Due to its simplicity, compatibility with different types of specimens, and ease of operation it is accepted by pathologists. ExM and its variants have been widely tested in different types of tissue specimens and microbiological specimens and yielded brilliant results.

Ensemble Deep-Learning-Based Prognostic and Prediction for Recurrence of Sporadic Odontogenic Keratocysts on Hematoxylin and Eosin Stained Pathological Images of Incisional Biopsies.

(1) Background: Odontogenic keratocysts (OKCs) are enigmatic developmental cysts that deserve special attention due to their heterogeneous appearance in histopathological characteristics and high recurrence rate. Despite several nomenclatures for classification, clinicians still confront challenges in its diagnosis and predicting its recurrence. This paper proposes an ensemble deep-learning-based prognostic and prediction algorithm, for the recurrence of sporadic odontogenic keratocysts, on hematoxylin and eosin stained pathological images of incisional biopsies before treatment. (2) Materials and Methods: In this study, we applied a deep-learning algorithm to an ensemble approach integrated with DenseNet-121, Inception-V3, and Inception-Resnet-V3 classifiers. Around 1660 hematoxylin and eosin stained pathologically annotated digital images of OKC-diagnosed (60) patients were supplied to train and predict recurrent OKCs. (3) Results: The presence of SEH (p = 0.004), an incomplete epithelial lining, (p = 0.023), and a corrugated surface (p = 0.049) were the most significant histological parameters distinguishing recurrent and non-recurrent OKCs. Amongst the classifiers, DenseNet-121 showed 93% accuracy in predicting recurrent OKCs. Furthermore, integrating and training the traditional ensemble model showed an accuracy of 95% and an AUC of 0.9872, with an execution time of 192.9 s. In comparison, our proposed model showed 97% accuracy with an execution time of 154.6 s. (4) Conclusions: Considering the outcome of our novel ensemble model, based on accuracy and execution time, the presented design could be embedded into a computer-aided design system for automation of risk stratification of odontogenic keratocysts.

Histopathologic Feature of Hyalinization Predicts Recurrence of Conventional/Solid Multicystic Ameloblastomas.

The histologic properties of tumors seem to affect their biological behavior, and the same holds good for solid multicystic ameloblastoma (SMA), a benign, locally destructive lesion. Hyalinization is one such histological factor that has been demonstrated to correlate with the biological behavior of neoplasms. The present study aimed to analyze the correlation between the severity of hyalinization (SOH) and the recurrence potential of SMAs. The study was performed on formalin-fixed, paraffin-embedded (FFPE) diagnosed archival cases of SMA, follicular SMA (n = 35) and plexiform SMA (n = 25). The cases were evaluated for SOH and scored from 0-3, and the correlation between SOH and recurrence was analyzed for statistical significance. The clinical parameters of the lesion were analyzed for statistical correlation with recurrence. The SOH significantly correlated with the recurrence of SMA (p = 0.001). The histologic type did not influence the biological behavior of SMA. The location of SMA in the body of the mandible (p = 0.036), multilocular radiolucency (p = 0.001) and root resorption (p = 0.002) also showed strong statistical correlation with recurrence. It is evident from the present study that hyalinization strongly correlates with the biological behavior of SMA. Future studies with advanced investigations could validate the presence of hyalinization and identify the origin of the hyalinized product in SMAs.

Erosive Effect of Acidic Beverages and Dietary Preservatives on Extracted Human Teeth-An In Vitro Analysis.

OBJECTIVES: Frequent consumption of acidic beverages and dietary preservatives in younger generation, diet-conscious (celebrities), and obese individuals have a rapid impact on demineralization of the teeth. An attempt was made to analyze the erosive potential of various acidic beverages. MATERIALS AND METHODS: One hundred and ninety extracted human permanent teeth were sectioned longitudinally, pre-weighed, randomly grouped, and placed in nine acidic beverages (200 mL) with predetermined pH, i.e., three commercially available fruit juices, three carbonated drinks, and three dietary preservatives. STATISTICAL ANALYSIS: The sectioned specimens (n = 10) were analyzed at time intervals of 12, 24, 48, and 96 days. Mean weight loss was calculated, and surface changes were assessed under a stereomicroscope. The demineralization pattern and microscopic changes were observed under a compound and polarizing microscope. One-way analysis of variance test followed by Tukey's post-hoc analysis was employed. RESULTS: Overall the maximum demineralizing effect was caused by vinegar and apple cider. In the fruit juices category, lemon juice induced significant changes, while in the carbonated drinks category Coca-Cola induced the maximum changes and in the category of food preservatives vinegar induced the maximum changes. Severe discoloration was seen with respect to Coca-Cola followed by Mountain Dew (carbonated drink). CONCLUSION: The present study is unique as three different types of microscopes have been employed and both dentin and enamel of permanent teeth have been analyzed. In addition, the effect of dietary preservatives on hard tissues was evaluated. Oral health educators can reinforce important practices such as decreasing the frequency of consumption and time duration of beverage contact with the teeth. Also, the use of mouth rinses and buffering agents after the consumption of dietary beverages can be advocated along with regular fluoride application for those who are regular consumers.

Subepithelial Hyalinisation Predicts Recurrence of Unicystic Ameloblastomas.

The inductive effect of hyalinisation and its influence on the biologic behaviour of ameloblastoma variants represent a scarcely researched domain of oral pathology. The complexity of the induction effects within the odontogenic apparatus, with the involvement of both ectodermal and mesodermal tissues, is responsible for diverse histopathological characteristics, hyalinisation being the major feature. The present study aims to deduce for the first time the correlation between the severity of hyalinisation (SOH) and recurrence in three unicystic ameloblastoma (UA) variants, namely, intra-luminal (UA-IL), luminal (UA-L) and mural (UA-M). Retrospectively diagnosed archival cases of UA-IL (n = 08), UA-L (n = 22) and UA-M (n = 30) were assessed for SOH and its correlation with recurrence. A subgroup comparison (between UA-IL/UA-L and UA-M) was also performed. The clinical parameters of the patients were also analysed from files for clinicopathological correlation with recurrence. Results: sub-epithelial hyalinisation (SEH) significantly correlated with the recurrence of UA-L and UA-M (p = 0.001). When the histologic types (UA-L and UA-IL vs. UA-M) were grouped and the correlation of SOH with recurrence was checked, it was observed that both groups (p = 0.001) showed strong statistical correlation. UA-M lesions with multilocular radiolucency (p = 0.001) also showed significant correlation with recurrence. SOH can be a reliable histological predictor of recurrence and of aggressive biologic behaviour in UA. The present study shows a significant association of hyalinisation with the biologic behaviour of UA. Further studies with immunohistochemical investigations could validate the presence of hyalinisation and identify the origin of the hyalinised product in UAs.

Efficacy of Csaba stain to demonstrate mature and immature mast cells in oral inflammatory lesions: An in-vitro study.

Background: Mast cells (MCs) are immune cells of the myeloid lineage and are present in connective tissues throughout the body. Mastocytosis regulates many physiologic processes and affects the pathogenesis of allergic conditions, anaphylactic reactions, autoimmune disorders and leukemias. Toluidine blue is commonly employed as a special stain for MCs that uniformly imparts blue color to both immature and mature cells. Csaba stain that distinguishes mature from immature MCs has not been widely explored. However, its use in differentiating mature and immature cells has not been reported in the literature. The identification of mature and immature MCs may provide a major clue for the diagnosis of any unrecognized systemic pathologies. Objectives: To evaluate the staining efficacy of Csaba stain, toluidine blue and Leishman's stains in the identification of MCsTo compare the effectiveness of Csaba stain, toluidine blue and Leishman's stains to identify MCs in inflammatory pathologic lesions. Materials and Methods: Fifty sections of inflammatory lesions were subjected to Csaba stain, toluidine blue and Leishman's stains each for studying MCs. The staining efficacy of the three stains was compared for parameters such as homogeneity, specificity, staining intensity, granular staining property and differentiation between mature and immature cells. Results: The Csaba, toluidine blue and Leishman's stains showed statistically insignificant P = 0.50, 0.95, 0.29, 0.48 for homogeneity, specificity, staining intensity and granular staining property respectively, but Csaba stain showed statistically significant P = 0.0001* in differentiating mature from immature cells. Conclusion: Csaba stain is more effective in differentiating mature from immature MCs compared to the other special stains.

Behavioral and Psychosocial Risk Factors that Lead Individuals to Initiate and/or Maintain Betel Quid Chewing in Developed and Developing Countries: A Systematic Review.

BACKGROUND: Betel quid (BQ) is the fourth most commonly used psychoactive substance and its use is highly prevalent among southeast Asian countries due to the influence of psychosocial, behavioral, and environmental factors. As a result, even young children and women are becoming addicted and find it difficult to achieve long-term abstinence. Systematic research addressing the influence of behavioral and psychosocial factors on BQ chewing is scarce, hence pointing to the need for understanding the interplay of these factors to develop tailored intervention strategies for BQ cessation. OBJECTIVE: The objective of this systematic review is to assess various behavioral and psychosocial risk factors from pre-existing literature that lead individuals to initiate and/or maintain BQ chewing in developed and developing countries. MATERIALS AND METHODS: The electronic retrieval systems and databases PUBMED [MEDLINE], EMBASE, SCOPUS, WEB OF SCIENCE, GOOGLE SCHOLAR, and SCIENCE DIRECT were searched independently by two authors for relevant articles from January 2011 to July 2021 using combinations of keywords. Primary studies published in English focusing on the behavioral and psychosocial risk factors for BQ chewing were included. RESULTS: Out of the initial 264 articles searched, 12 articles met the selection criteria. Included studies addressed the influence of behavioral and psychosocial factors toward the initiation and/or maintenance of BQ chewing. Included studies reported that dependent BQ was not able to quit BQ chewing due to habituation, addiction, and withdrawal symptoms. Few social BQ chewers in Southeast Asian countries were not willing to initiate quitting as they considered AN as an important social and cultural identifier and claimed that they could not resist chewing in a peer group, and it has become part of their social life. CONCLUSION: In the future, studies should take into consideration behavioral and psychosocial risk factors, which are major barriers toward successful quitting. These factors should be integrated into areca nut cessation guidelines, and there is a need to develop more comprehensive culture-specific intervention approaches to achieve long-term abstinence.

SARS-CoV-2 infection in heart transplant recipients: a systematic literature review of clinical outcomes and immunosuppression strategies.

The impact of SARS-CoV-2 infection on heart transplant recipients is unknown. Literature is limited to case reports and series. The purpose of this study is to identify the clinical features, outcomes, and immunosuppression strategies of heart transplant recipients with COVID-19 infection. A systematic review was conducted using the search term "Coronavirus" or COVID," "SARS-CoV-2," "cardiac transplantation," and "heart transplant." Case reports and retrospective studies were gathered by searching Medline/PubMed, Google Scholar, CINAHL, Cochrane CENTRAL, and Web of Science. Thirty-three articles were selected for review. We identified 74 cases of SARS-CoV-2 infection in heart transplant and heart-kidney transplant recipients. The mean age was 60.5 ± 15.8 years, and 82.4% were males with median time from transplant of 6.5 years. Commonest symptoms were fever, cough, and dyspnea, but new left ventricular (LV) dysfunction was rare. Leukocytosis, lymphopenia, elevated inflammatory markers, and bilateral ground-glass opacities were common. Mortality was high, with particularly poor survival in patients who required intensive care unit (ICU) admission and older patients. Immunosuppression involved discontinuation of antimetabolites and steroids. COVID-19 infection in heart transplant (HT) recipients presents similarly to the general population, but new onset of LV dysfunction is uncommon. Immunosuppression strategies include increase in corticosteroids and discontinuation of antimetabolites.

P-Cadherin and WNT5A expression in assessment of lymph node metastasis in oral squamous cell carcinoma.

BACKGROUND: Oral cancer progression is a multi-step process in which adhesion molecules play a pivotal role in the development of recurrent, invasive, and distant metastasis. The aim of this study was to adopt prognostic biomarkers to assess the lymph node metastasis of OSCC that will facilitate in deciding the treatment modality by the surgeons. OBJECTIVES: The objectives of the study were to assess the biological behaviour of OSCC by correlating the expression levels of P-Cadherin and WNT5A immunohistochemically. METHODS: A total of 60 selected OSCCs cases (lymph node metastasis n = 30, non-metastatic n = 30) and 10 normal healthy controls were quantitatively and qualitatively analysed by immunohistochemistry for P-Cadherin and WNT5A. A survival analysis was also performed. RESULTS: The expression levels of P-Cadherin and WNT5A in OSCC groups were statistically significant between metastatic and non-metastatic groups (p < 0.001). P-Cadherin and WNT5A expression in metastatic (lymph node metastasis) and non-metastatic cases showed a significant correlation coefficient of 0.753 at (p < 0.01). The present study also found that the aberrant expression (high) of P-Cadherin was associated with diminished survival of patients with metastatic OSCC. CONCLUSION: The present study demonstrated the aberrant expression of P-Cadherin and WNT5A could serve as important prognosticator in OSCC. CLINICAL RELEVANCE: P-Cadherin and WNT5A could be used as significant predictors of disease outcome.

Deep Learning-Based Microscopic Diagnosis of Odontogenic Keratocysts and Non-Keratocysts in Haematoxylin and Eosin-Stained Incisional Biopsies.

BACKGROUND: The goal of the study was to create a histopathology image classification automation system that could identify odontogenic keratocysts in hematoxylin and eosin-stained jaw cyst sections. METHODS: From 54 odontogenic keratocysts, 23 dentigerous cysts, and 20 radicular cysts, about 2657 microscopic pictures with 400× magnification were obtained. The images were annotated by a pathologist and categorized into epithelium, cystic lumen, and stroma of keratocysts and non-keratocysts. Preprocessing was performed in two steps; the first is data augmentation, as the Deep Learning techniques (DLT) improve their performance with increased data size. Secondly, the epithelial region was selected as the region of interest. RESULTS: Four experiments were conducted using the DLT. In the first, a pre-trained VGG16 was employed to classify after-image augmentation. In the second, DenseNet-169 was implemented for image classification on the augmented images. In the third, DenseNet-169 was trained on the two-step preprocessed images. In the last experiment, two and three results were averaged to obtain an accuracy of 93% on OKC and non-OKC images. CONCLUSIONS: The proposed algorithm may fit into the automation system of OKC and non-OKC diagnosis. Utmost care was taken in the manual process of image acquisition (minimum 28-30 images/slide at 40× magnification covering the entire stretch of epithelium and stromal component). Further, there is scope to improve the accuracy rate and make it human bias free by using a whole slide imaging scanner for image acquisition from slides.

Role of cancer stem cells in head-and-neck squamous cell carcinoma - A systematic review.

Targeting cancer stem cell (CSC) subpopulation within the tumor remains an obstacle for specific therapy in head-and-neck squamous cell carcinoma (HNSCC). Few studies in the literature describe a panel of stem cell makers, however a distinct panel has not been put forth. This systematic review aims to enhance the knowledge of additional markers to accurately relate their expression to tumorigenesis, metastasis, and therapy resistance. Databases, including PubMed, Google Scholar, Ebsco, and Science Direct, were searched from 2010 to 2017 using various combinations of the following keywords: "Stem cell markers in HNSCC" and "chemoresistance and radioresistence in HNSCC." Original experimental studies (both in vitro and in vivo) published in English considering stem cell markers in HNSCC, were considered and included. We excluded articles on tumors other than HNSCC, reviews, editorial letters, book chapters, opinions, and abstracts from the analyses. Forty-two articles were included, in which 13 types of stem cell markers were identified. The most commonly expressed CSC markers were CD44, aldehyde dehydrogenase, and CD133, which were responsible for tumorigenesis, self-renewal, and therapy resistance, whereas NANOG, SOX-2, and OCT-4 were involved in metastasis and invasion. Identification of an accurate panel of CSC markers is the need of the hour as nonspecificity of the current markers poses a problem. Further studies with a large sample size would help validate the role of these CSC markers in HNSCC. These CSC proteins can be developed as therapeutic targets for HNSCC therapy, making future treatment modality more specific and effective.

A Histopathology-based Assessment of Biological Behavior in Oral Hyalinizing Extraosseous Lesions by Differential Stains.

AIM: The assessment of hyalinization to determine aggressive behavior in oral pathological lesions is a scarcely researched field that requires further exploration. The current study aims to predict the biological behavior of oral hyalinizing extraosseous lesions (OHEOL) by employing four differential stains with clinicopathologic correlation. MATERIALS AND METHODS: The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of salivary gland tumors (SGTs) (n = 13), benign soft tissue (BST) lesions (n = 24), and oral submucous fibrosis (OSMF) (n = 53). The hematoxylin and eosin-stained sections were analyzed for the severity of hyalinization (SOH). Differential stains periodic acid Schiff (PAS), Alcian blue, safranin O, and picrosirius red with polarizing microscopy were used to assess the components of hyalinized tissue. The SOH was correlated with differential staining characteristics and clinicopathologic features to analyze possible correlation with aggressive potential in BST, advancement of disease in OSMF, and recurrence in SGT. RESULTS: Intensity of picrosirius red stain significantly correlated with SOH of SGTs (p = 0.044). The intensity of PAS stain (p = 0.040), picrosirius red polarizing greenish-yellow color (p = 0.002), and pattern of distribution of picrosirius red (p = 0.023) significantly correlated with recurrence of SGTs. The intensity of differential stains increased with the SOH in BST lesions indicating their correlation with SOH. The intensity (p = 0.008) and pattern (p = 0.010) of Alcian blue staining and intensity of safranin O stain (p = 0.003) significantly correlated with SOH in OSMF. Picrosirius red polarizing color reddish and yellowish red (p = 0.002) significantly correlated with SOH distinguishing early and advanced OSMF. CONCLUSION: Picrosirius red and PAS stains are reliable indicators of SOH and recurrence potential in SGT. Alcian blue, safranin O, and picrosirius red polarizing colors enable detection of SOH and accurately distinguish early from advanced OSMF. CLINICAL SIGNIFICANCE: SOH can be considered as a histological predictor of aggressive biologic behavior in OHEOL. These findings will result in appropriate management protocols.

Natural stain (Kumkum) formulated by the extract of Curcuma aromatica and slaked lime in histostaining of oral tissues: An observational study.

BACKGROUND: The pharmacological actions of Curcuma aromatica (wild turmeric) such as anti-inflammatory, antitumor, antifungal, antimicrobial and wound healing have been recognized since ages. However, its role as a natural histological stain has not been explored till date. AIM: To evaluate the efficacy of natural substance-Kumkum prepared from the extract of C. aromatica and slaked lime in staining the biopsied oral tissues. MATERIALS AND METHODS: A cohort study that used 60 formalin fixed paraffin embedded soft and hard tissue specimens from institutional archives were subjected to sectioning and stained using Kumkum and hematoxylin and eosin (H and E). The slides were evaluated for their staining efficacy and results were statistically analyzed using Wilcoxon signed-rank test and Independent 't' test. RESULTS: The mean of the overall parameters assessed for staining efficacy did not show statistically significant difference between the study groups in normal and pathological specimens for tooth (P = 0.410 and 0.484), bone (P = 0.133 and 0.157) and soft tissues (P = 0.186 and 0.113), respectively. This suggests that Kumkum staining efficacy is equivalent to that of routine H and E for oral tissues. Structures such as dentinoenamel junction, dentinal tubules, incremental lines of cementum, reversal and resting lines, osteocytic canaliculi, mature and immature bone could be appreciated better in Kumkum stained slides, thereby rendering a special staining property to Kumkum stain. CONCLUSION: To our knowledge, this study is the first of its kind to have used Kumkum stain obtained from C. aromatica for the differentiation of the components of tooth, bone and soft tissue structures in histostaining of oral tissues. The naturally prepared Kumkum stain possesses dual staining property both in routine and differential staining. This facilitates diagnosis of fibro-osseous lesions, bony, collagen and muscular pathologies. The natural stain also finds application in forensic odontology for age estimation.

A Histopathology-based Assessment of Biological Behavior in Oral Hyalinizing Odontogenic Tumors and Bone Lesions by Differential Stains.

AIM: Odontogenic tumors (OTs) and bone lesions of the oral cavity present diverse histological features and varying clinical behavior that makes predicting their biologic behavior difficult. The research undertaken in the current study aims to predict the biological behavior of oral hyalinizing odontogenic and bone lesions (OHO-BL) for the first time by employing four differential stains with clinicopathologic correlation. MATERIALS AND METHODS: The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of OTs (n = 53) and bone lesions (n = 10). The severity of hyalinization (SOH) was assessed from stained tissue sections. Polarizing microscopy was used to analyze hyalinization in tissues stained with differential special stains, namely periodic acid-Schiff (PAS), Safranin-O, Alcian Blue, and Picrosirius red. SOH was also analyzed for possible correlation with recurrence and clinicopathologic correlation in OHO-BL. RESULTS: Intense staining was observed with PAS, Alcian Blue, and Safranin-O in OTs with increased SOH with a statistical significance. Polarizing greenish yellow color correlated significantly with the recurrence potential of the OT group. Recurrence in individual lesions of the OT group showed a statistically significant association with SOH. Such individual correlation was not observed in bone diseases. CONCLUSION: PAS, Alcian Blue, Safranin-O, and Picrosirius red are reliable stains to assess hyalinization in OHO-BL. Picrosirius red-polarizing microscopy is a dependable tool for identifying recurrent odontogenic lesions. CLINICAL SIGNIFICANCE: SOH can be considered a histological predictor of aggressive biologic behavior in oral hyalinizing odontogenic lesions that can enable the surgeon to arrive at an appropriate management protocol.

Proliferative Verrucous Leukoplakia Revisited: A Retrospective Clinicopathological Study.

(1) Objective: To review the criteria proposed by Cerero-Lapiedra et al. and to retrospectively identify the under-diagnosed disease in patients diagnosed with proliferative verrucous leukoplakia. (2) Materials and methods: In this study, we included patients who were diagnosed with leukoplakia (histological label consistent with the clinical diagnosis, n = 95), and cases with a final diagnosis within the spectrum of proliferative verrucous leukoplakia (n = 110) as defined by Batsakis et al. We applied the criteria proposed by Cerero-Lepiedra et al. to screen for the possible cases of proliferative verrucous leukoplakia. (3) Results: Although many of our patients satisfied specific isolated criteria, only 11 cases satisfied specific combinations of the guidelines to satisfy a diagnosis of proliferative verrucous leukoplakia. However, due to the lack of follow-up data, the disease is not confirmed in these 11 cases. (4) Conclusion: A limited number of cases of proliferative verrucous leukoplakia were diagnosed using the criteria given by Cerero-Lapiedra et al. The true natural history of the disease could not be studied due to the lack of follow-up data. (5) Clinical relevance: Proliferative verrucous leukoplakia presenting as hyperkeratosis or mild epithelial dysplasia are often not followed up, and they subsequently transform into carcinoma. Thus, clinicians must be vigilant whenever they encounter leukoplakia, especially with multifocal presentations. In such cases, the follow-up data are the key to understanding the true nature of the disease entity.

Lipid metabolism in cancer: A systematic review.

Preclinical studies and clinical trials have emphasized the decisive role of lipid metabolism in tumor proliferation and metastasis. This systematic review aimed to explore the existing literature to evaluate the role and significance of the genes and pathways most commonly involved in the regulation of lipid metabolism in cancer. The literature search was performed as per Preferred Reporting Items for Systematic Reviews and Meta-analyses. Approximately 2396 research articles were initially selected, of which 215 were identified as potentially relevant for abstract review. Upon further scrutiny, 62 of the 215 studies were reviews, seminars, or presentations, and 44 were original study articles and were thus included in the systematic review. The predominant gene involved in lipid metabolism in cancer was stearoyl-coenzyme A desaturase 1 (SCD1), followed by fatty acid synthase (FASN). The pathway most commonly involved in lipid metabolism in cancer was the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, followed by the mitogen activated protein kinase (MAPK) pathway. SCD1 and FASN play significant roles in the initiation and progression of cancer and represent attractive targets for potentially effective anti-cancer treatment strategies. The regulation of cancer metabolism by the Akt kinases will be an interesting topic of future study.

Hyalinization as a histomorphological risk predictor in oral pathological lesions.

BACKGROUND: Hyalinization is a process of conversion of stromal connective tissue into a homogeneous, acellular translucent material. Nevertheless, hyalinization could provide insights into the biologic behaviour and prognosis of pathological lesions. Few studies with limited sample size have intended to assess the correlation of hyalinization and biologic behaviour in oral lesions. AIM: The current review aims to comprehensively appraise the mechanism of hyalinization in pathological oral hyalinizing lesions (OHL) and its clinical implications with emphasis on differential stains employed. METHODS: An electronic search was performed in the PubMed database (from year 2000-2020) using the keywords "special stains in oral hyalinizing lesions", "significance of hyalinization in oral lesions" and "hyalinization and biologic behaviour". Original research articles analyzing the effect of hyalinization on biologic characteristics of the lesion were evaluated in this review. Narrative review articles that provided insights into the mechanism of hyalinization and maturity of collagen fibers were also considered for analysis. CONCLUSION: The presence of hyalinization does seem to have a significant effect on the biologic behaviour of pathological lesions. There is substantial scope to further investigate the process of hyalinization on larger samples and its correlation with the aggressive behaviour of OHLs. Special stains and advanced investigations such as immunohistochemistry for stromal markers would define the nature of hyalinized material and validate the correlation. CLINICAL SIGNIFICANCE: The prediction of the biologic behaviour of a lesion established through assessment of hyalinization would prevent unwanted over or under treatment leading to a better prognosis.