Srikant Natarajan

Prognostic Efficacy of Nuclear Morphometry at Invasive Front of Oral Squamous Cell Carcinoma: An Image Analysis Microscopic Study

Background. Oral cancer is revisited on a pathologist perspective with advanced imaging technique. Objective. The present study assessed the new malignancy grading system at tumor proper (TP) and Brynes grading system at invasive tumor front (ITF), morphometric features using software, to clarify their associations with prognosis of oral cancers. Methods. Histologically confirmed oral squamous cell carcinoma (OSCC) with 5-year follow-up was assessed morphometrically using image analysis at TP and ITF, correlated with the prognosis of patient. Results. On comparison of grading systems, a moderate agreement between both (Bryne and Anneroth) was seen. Among all histological parameters, we noted inverse correlation between degree of mitosis at invasive front and decrease in lymphoplasmacytic infiltrate at ITF with increase rate of recurrence and event of death. An increase in nuclear area, diameter, and perimeter along with decrease circularity in advancing OSCC was seen. Correlation of parameters showed higher values for maximum nuclear diameter, perimeter, and circularity at TP and ITF with recurrence. Conclusion. This study, while limited in sample size, concluded that a combined assessment of clinical TNM staging, histopathological grading system {excluding the parameter “mitotic activity” (due to its inverse relation)}, and nuclear morphometry at the ITF are better prognosticators. This combination proved to be an accurate predictive factor in eliciting the varied molecular characteristics of tumor heterogeneity.

A novel technique to assess chromatin texture using pixel optical densitometry in oral squamous cell carcinoma

Background: The early detection of neoplasia is the prime aim of a diagnostician. Altered chromatin distribution is the earliest microscopic change observed in malignant transformation making it a valuable morphometric parameter. This study was aimed to assess and correlate the progressive changes in chromatin texture from normal to varying grades of premalignancies to malignancy of the oral cavity. Methods: Thirty‐four archival tissue specimens categorized as normal buccal mucosa (5), low‐grade epithelial dysplasia (11), high‐grade epithelial dysplasia (7), and squamous cell carcinoma (11) were stained with Feulgen reaction. Pixel optical densitometry histograms were obtained from analysis of an average of 300 cells/case using ImageJ software. Nine histogram curve characteristics (including area under the curve, area integer percentage, center, centroid) were then analyzed statistically for differences between the four groups. Results: Area integer percentage (P = 0.002), center (P = 0.038), and centroid (P = 0.021) were statistically significant within the four groups The parameters showed a dip in their value from normal to low‐grade dysplasia but showed a steady increase in high‐grade dysplasia and carcinoma. Conclusion: The optical density characteristics had the ability to differentiate the progression of neoplastic changes by extracting information of the chromatin distribution not quantifiable by routine microscopy. The early clumping of chromatin in the periphery in low‐grade dysplasia followed by progressively increasing chromatin aggregates in high‐grade dysplasia and carcinoma correlated well with the shift observed in the histogram center and centroid. The novel pixel optical densitometry technique efficiently predicted malignant transformation. Microsc. Res. Tech. 75:1119–1123, 2012.

Morphometric Analysis of Nuclear Features and Volume-Corrected Mitotic Index in the Prognosis of Oral Squamous Cell Carcinoma

Abstract In the present study, an objective and reproducible evaluation of mitotic activity and nuclear morphometric factors was attempted in 30 patients of oral squamous cell carcinoma with a view to predicting local relapse and survival. Various nuclear parameters and volume-corrected mitotic index were calculated and compared with the recurrence and death of the study group (n = 30). Volume-corrected mitotic index (M/V, index) was the single best prognosticator for recurrence of oral squamous cell carcinoma (p = 0.008 for recurrence; p=0.015 for death). The combination of M/V and SD of nuclear area (forward stepwise regression) was a more efficient and better predictor of survival (Log rank test, Kaplan-Meiers survival analysis; χ 2 = 17.46, p=0.00001). The present study proved the effectiveness of the M/V index ill predicting the biological behavior and the outcome of oral squamous cell carcinoma patients.

Driving the Mineral out Faster: Simple Modifications of the Decalcification Technique.

INTRODUCTION Quicker decalcification is essential for faster diagnosis of hard tissue pathology. Heat and mechanical agitation are known to hasten decalcification. AIM To compare the rate of decalcification, cellular and staining characteristics of decalcified specimens of bone and teeth by using the conventional method (10% formal formic acid), heating to 45(o)C and by physical agitation with magnetic stirrer. MATERIALS AND METHODS Weight-matched samples of caprine-origin bone (n=15) and teeth (n=15) were decalcified using three methods namely: a) Gooding and Stewarts fluid; b) Gooding and Stewarts fluid heated to 45(o)C for 6 hours daily; and c) Gooding and Stewarts fluid agitated using a magnetic stirrer for 6 hours daily. Non-lesional skin tissue samples were placed along with each specimen. End point of decalcification (chemical test) was noted; 4 micron sections were taken and stained with H&E. STATISTICAL ANALYSIS Differences in rate of decalcification and staining characteristics were assessed by Kruskal Wallis test and chi-square test respectively. RESULTS Hard tissues decalcified faster with stirring and heating methods. The amount of osteocyte retraction noted in bone was significantly reduced in the stirring method. In tooth specimens, modified techniques resulted in poorer nuclear-cytoplasmic contrast of pulp cells. Heating affected the odontoblast layer. Soft tissues exhibited higher eosinophilia in stirring and conventional methods, whereas nuclear-cytoplasmic contrast and chromatin staining was poorest in heating and conventional methods. CONCLUSION Physical agitation of decalcifying fluid may be recommended while maintaining satisfactory quality of tissue morphology and staining.

Tumour-Associated Tissue Eosinophilia in Oral Squamous Cell Carcinoma- A Boon or a Bane?

INTRODUCTION The infiltration of tumour stroma by eosinophils, Tumour-Associated Tissue Eosinophilia (TATE) is known to modulate the evolution of Oral Squamous Cell Carcinoma (OSCC). Identification of eosinophils in the inflammatory stroma has been proven to be an important factor in prognostication of malignant tumours including cancers of mouth, oesophagus, larynx, pharynx, breast, lung, intestine and genitourinary tract. AIM Our study aimed to assess the role of TATE as a prognosticator in OSCC as visualized by Haematoxylin and Eosin (H&E) and congo red staining. MATERIALS AND METHODS Thirty histologically-proven cases of OSCC were retrieved from the archives of Department of Oral Pathology, Manipal College of Dental Sciences, Mangalore, Manipal University, Karnataka, India. Two serial sections of 4μm thickness were made and subjected to routine staining with H&E and modified congo red staining, where eosinophil granules stained red and nuclei stained blue. In 40x magnification, 10 HPF at invasive tumour front were assessed for counting eosinophils by placing a 49 square grid (measuring 0.0289 sq mm). STATISTICAL ANALYSIS The TATE was compared with the prognosticators using Mann-Whitney U-test. The grades of carcinoma were correlated with TATE using Kruskal-Wallis test followed by Post-hoc Bonferronis correction. Agreement of the number of eosinophils counted in the two staining techniques (H&E and Congo red) in OSCC was achieved using interclass correlation coefficient, and Friedmans test. A value of p< 0.05 was considered statistically significant. RESULTS Our results showed that tissue eosinophil counts were higher in well-differentiated cases of OSCC, cases with lymph node involvement, decreased survival, without margin involvement and in cases that did not recur. H&E stain showed significantly better visualization of eosinophils resulting in higher eosinophil counts than when seen with Congo red (p=0.008). CONCLUSION Thus, TATE can be used as a surrogate marker in prediction of survival and recurrence in OSCC. H&E proved to be a better stain for evaluation of eosinophils.

Palatal Swelling in a Patient Suffering from Filariasis

Sclerosing mucoepidermoid carcinoma (SMEC) is a distinct but an uncommon salivary gland neoplasm with only 19 cases reported in English literature till date. Densely collagenous sclerotic stroma, resemblance to other benign lesions and rarity of this tumour often makes the diagnosis of SMEC challenging. Here we report a case of SMEC in a 73-year old female patient suffering from filariasis. Clinical, radiological, fine needle aspiration biopsy, gross and histopathological features are discussed with detailed review of literature and probable pathogenesis.

rac-2-Hy­droxy-2-(2-oxocyclo­pent­yl)-1H-indene-1,3(2H)-dione

In the title compound, C(14)H(12)O(4), the indene unit is essentially planar [r.m.s. deviation = 0.0309 (1) Å] and the cyclo-penta-none ring adopts a twist form. In the crystal, mol-ecules are joined via pairs of O-H⋯O hydrogen bonds into centrosymmetric dimers.

Understanding patterns of invasion: a novel approach to assessment of podoplanin expression in the prediction of lymph node metastasis in oral squamous cell carcinoma.

Cellular motility is considered to be central to the process of metastasis, and podoplanin expression can be explored as a prospective link, owing to its ability to modulate the actin cytoskeleton. We aimed to evaluate the tumoral expression of D2‐40 (monoclonal antibody against podoplanin) in pathologically neck‐node‐negative/positive cases (pN0/N+) to characterise the pattern of invasion, potentially explaining the role of various patterns of invasion in causing tumour metastasis.

Surgical management of complex odontoma associated with agenesis of a molar

Odontomas are mixed odontogenic tumors composed of mineralized tissue of ectomesenchymal origin. These are usually detected in routine radiographs and may have a variable etiology such as local trauma, genetic mutation, or infection. They are classified into two main types: complex and compound. Complex odontomas are rare and they rarely erupt into the oral cavity as they lack periodontal ligament. This is the report of one such case of complex odontoma where an 11-year-old girl reported with an asymptomatic swelling, in relation to the left angle of the mandible. The case report highlights the clinical and radiographic findings, investigations done to arrive at the diagnosis and the treatment rendered. This report is of clinical importance because of its rare association with the agenesis of a molar tooth.

Odontogenic Cyst with Verrucous Proliferation Exhibiting Melanin Pigmentation

Verrucous proliferation arising from odontogenic cysts is a rare entity. We report an unusual case of an infected odontogenic cyst with verrucous proliferation and melanin pigmentation in a 13-year-old male patient who presented with an intraoral swelling in relation to impacted teeth 26 and 27. The enucleated lesion was diagnosed as an odontogenic keratocyst and the patient died within two years of presentation due to multiple recurrences. The clinical, radiological, and microscopic features of the lesion are presented with an attempt to discuss the etiopathogenesis. The case hereby reported is uncommon with only eight cases reported in the literature.