Seema Hallikerimath

Activation of latent TGF‐β1 by low‐power laser in vitro correlates with increased TGF‐β1 levels in laser‐enhanced oral wound healing

The term Laser “Photobiomodulation” was coined to encompass the pleiotropic effects of low‐power lasers on biological processes. The purpose of this study was to investigate whether transforming growth factor (TGF)‐β had a role in mediating the biological effects of low‐power far‐infrared laser irradiation. We assayed for in vitro activation using various biological forms of cell‐secreted, recombinant, and serum latent TGF‐β using the p3TP reporter and enzyme‐linked immunosorbent assays. We demonstrate here that low‐power lasers are capable of activating latent TGF‐β1 and ‐β3 in vitro and, further, that it is capable of “priming” these complexes, making them more amenable to physiological activation present in the healing milieu. Using an in vivo oral tooth extraction‐healing model, we observed an increased TGF‐β1, but not β3, expression by immunohistochemistry immediately following laser irradiation while TGF‐β3 expression was increased after 14 days, concomitant with an increased inflammatory infiltrate. All comparisons were performed between laser‐irradiated wounds and nonirradiated wounds in each subject essentially using them as their own control (paired T‐test p<0.05). Low‐power laser irradiation is capable of activating the latent TGF‐β1 complex in vitro and its expression pattern in vivo suggests that TGF‐β play a central role in mediating the accelerated healing response.

Evaluation of myofibroblasts in oral submucous fibrosis: correlation with disease severity

BACKGROUND Oral submucous fibrosis (OSMF) is a chronic debilitating disease and a premalignant condition of the oral cavity characterized by generalized submucosal fibrosis. Myofibroblasts are contractile cells expressing α-smooth muscle actin (α-SMA) and are considered primary producers of extracellular matrix after injury. Their accumulation has been established as a marker of progressive fibrosis in organs like lungs, liver, kidney and skin. This study aims to evaluate the presence of myofibroblasts in various histological stages of OSMF. MATERIALS AND METHOD Seventy cases of OSMF, which were further categorized histologically into early (35 cases) and advanced (35 cases), were subjected to immunohistochemistry using α-SMA antibody for detection of myofibroblasts. Fifteen normal oral mucosa specimens were also stained as controls. RESULTS The number of α-SMA-stained myofibroblasts in OSMF was significantly increased when compared to that of the normal controls (P<0.001). Additionally, a statistically significant increase in the myofibroblasts population between early and advanced stages was observed (P=0.000). CONCLUSIONS Our results corroborate the possibility that OSMF actually represents an abnormal healing process in response to chronic mechanical and chemical irritation because of areca nut chewing as demonstrated by the increased incidence of myofibroblasts in this disease. Furthermore, the progressive increase in myofibroblasts from early to advanced stages suggests their potential use as markers for evaluating the severity of OSMF.

Genetic polymorphism of drug metabolizing enzymes (GSTM1 and CYP1A1) as risk factors for oral premalignant lesions and oral cancer

AIMS Polymorphisms in the genes that code for metabolic enzymes involved in either the activation (Phase I) or detoxication (Phase II) of chemical carcinogens in tobacco, may alter expression or function of carcinogenic compounds and hence alter risk of oral cancer. The present study investigates whether polymorphisms at CYP1A1 and GSTM1 gene loci act as risk factors for oral precancerous lesions and cancer. METHODS For the present study, histopathologically confirmed cases of 90 oral precancerous lesions, 150 oral squamous cell carcinoma (SCC) and 150 control subjects were selected. Polymerase chain reaction and restriction fragment length polymorphism were performed using DNA from blood samples to determine the polymorphic genotypes at CYP1A1 and GSTM1 loci. RESULTS CYP1A1 C (m2/m2) genotype conferred a 12.0 fold-increased risk (OR=12.0; 95% CI, 2.40-60.05) to oral SCC. GSTM1 null showed no significant association but the frequency was higher in oral SCC cases. Patients with genotype C and/or GSTM1 deficiency developed carcinoma after less tobacco consumption than those of other genotypes though the difference was not statistically significant. The frequency of the combined genotypes C and GSTM1 null was found to be 14% among oral SCC patients. On comparing the susceptibility of intraoral sites it was found that in the majority of cases (64%) in the study groups they were the buccal mucosa. CONCLUSION Hence it was concluded that metabolic enzymes reported in the present study: CYP1A1 significantly alter oral cancer risk. GSTM1 null and CYP1A1 C (m2m2) show a predisposition to premalignant lesions and cancer of the buccal mucosa than other sites.

Age estimation by pulp/tooth area ratio in canines: Cameriere's method assessed in an Indian sample using radiovisiography

Age estimation of an individual whether living or dead is an intimidating task in forensic investigations. Since teeth are more resistant to most peri- and post-mortem changes, they are frequently used for identification and age estimation when skeletal remains are in poor condition. However, most methods are destructive and warrant extraction of teeth which is not feasible in living individuals. Camerieres et al. put forth a radiographic method of age estimation by pulp to tooth area ratio (AR) in canines and revealed a linear regression between age and the AR. In the present study, we estimated the AR in 456 canines (upper, lower and both) in an Indian sample (114 males and 114 females) using radiovisiography technique. Linear regression equations were derived for upper canine, lower canine and both using the AR to estimate chronological age. Additionally, the efficacy of these equations was also evaluated in younger age group (<45 years). The formulas derived, i.e., age=96.795-513.561x(1) (Eq. (1)) for upper canine, age=88.308-458.137x(2) (Eq. (2)) for lower canine and age=99.190-283.537x(1)-306.902x(2)+400.873x(1)x(2) (Eq. (3)) for both the canines were applied to predict the chronological age. The mean value of residuals using these regression equations ranged from 4.28 to 6.39 years with upper canine equation generally giving a precise result. When these equations were applied for younger ages (<45 years), the regression equation derived from both canines gave a better result (mean residual 2.70 years). Overall these equations were better able to predict the age in younger ages, i.e., up to 45 years.

Reverse transcriptase polymerase chain reaction study to evaluate dissemination of cancer cells into circulation after incision biopsy in oral squamous cell carcinoma.

BACKGROUND Tissue manipulation by incisions, radiotherapy, and palpation may lead to dissemination of cancer cells into circulation. Circulating cancer cells in blood play a central role in metastatic process. Their numbers can be very small and for their detection,reverse transcriptase polymerase chain reaction (RT-PCR) has been successfully used in this study. MATERIALS AND METHODS To examine whether cancer cell dissemination results from incision biopsy, we tried to detect oral squamous cell carcinoma (OSCC) cells in the peripheral blood sample before and after incision biopsy by CK19 RT-PCR. The study group consisted of 25 OSCC patients and the control group consisted of five patients with oral submucos fibrosis and five with leukoplakia. Five ml of blood collected before and twice (15 and 30 min) after incision were used for CK19 RT-PCR. RESULTS Four (16%) of 25 cases of OSCC were positive for CK19 transcripts in their peripheral blood drained 15 min after incision. CK19 transcripts were not detected in the control group. CONCLUSION Surgical invasion, in the form of incisional biopsy, causes dissemination of cancer cells into circulation, resulting in increased risk of metastasis.

Assessment of frontal sinus dimensions to determine sexual dimorphism among Indian adults.

Background: Sex identification of unknown individuals is important in forensic sciences. At times when only skull remains are found and other means of identification fails, radiographs of frontal sinus can be used for identification. Frontal sinus morphology is unique to individual and can be used effectively in person identification; whereas its use in determining sexual dimorphism is limited. Aim: To determine sexual dimorphism among Indians by evaluating frontal sinus pattern using postero-anterior radiograph. Materials and Methods: The right and left areas, maximum height and width of frontal sinus were determined in 300 digital postero-anterior view radiographs obtained from 150 males and 150 females aged between 18-30 years. The measurements were carried out by transferring the image to Adobe® Photoshop® CS3 extended. Comparison of values were done using students t-test and accuracy of sex determination was assessed through Logistic regression analysis. Results and Conclusions: One hundred and forty seven males and 142 females show presence of frontal sinus with seven individuals showing unilateral/bilateral absence of frontal sinuses. The mean values of the frontal sinus height, width and area are greater in males. Right frontal sinus is larger than the left sinus in both the sex. The mathematical model based on logistic regression analysis gives an average concordance index for sex determination of 64.6%. Thus, frontal sinus provides average accuracy in sex determination among Indian population. This may be due to its greater variation in morphology.

Sex assessment using odontometry and cranial anthropometry: evaluation in an Indian sample

Crania and teeth are considered to be useful adjuncts for sex assessment and in construction of a postmortem profile, however, there is very little information regarding the relationship between tooth and cranial size. The purpose of this study was to demonstrate the extent of sexual dimorphism of teeth and cranial size in an adult Indian population and their potential in sex estimation using logistic regression analysis. The sample consisted of 200 subjects (96 males and 104 females; age range of 18–30 years) of Indian origin. Cranial anthropometric measurements i.e. maximum head length and head breadth were measured and cephalic index was calculated. Tooth size (maximum mesiodistal and buccolingual dimensions) was measured for all the permanent teeth of the right side of the maxillary and mandibular arches, except the third molars. To ascertain the usefulness of absolute measurements of crania and teeth and the combination of both these parameters in sex prediction, logistic regression analysis was done. The cranial anthropometric measurements gave a sex assessment accuracy ranging from 53.5 to 79.9%, with head length giving the best accuracy. The sex classification accuracy of the odontometric measurements ranged from 61.5 to 76%, with combination of maxillary and mandibular teeth giving better accuracy. The use of both these parameters together vastly improved the accuracy to 88.4%. This study demonstrates that cranial anthropometry along with odontometry could be used fordetermining the gender of adult Indians in a forensic context. Among all the parameters, head length gave a superior sex prediction alone (79.9%) as well as in combination with odontometry (88.4%).

Tumor Budding Is an Independent Prognostic Factor for Prediction of Lymph Node Metastasis in Oral Squamous Cell Carcinoma

Objective. Despite the enormous advances in diagnostic and management modalities of oral squamous cell carcinoma (OSCC), the mortality rates have remained stagnant with a 5-year survival rate of <50% challenging the available methods of prognostic assessment. Presence of tumor budding has been associated with aggressive behavior and is correlated with lymph node metastasis, recurrence, distant metastasis, and decreased survival in several cancers. However, the prognostic significance of this apparently simple to evaluate parameter is sparse in OSCC. Methods. A total of 75 cases of surgically excised OSCC were analyzed for tumor budding along with other clinicopathologic parameters. Tumor budding was graded as high and low intensity based on presence and absence of ≥10 or <10 budding foci in hematoxylin and eosin–stained sections. An association between the clinicopathological parameters, lymph node metastases with the budding index was examined using univariate and multivariate analyses. Results. Tumor budding was evident in 89% of cases with around 45.3% of the cases demonstrated high-intensity budding. High-intensity tumor budding was significantly associated with lymph node metastasis and depth of invasion. Multivariate analysis demonstrated that tumor budding and depth of invasion were significant independent predictors for lymph node metastasis. Conclusion. Tumor budding is frequently encountered histologic marker in OSCC. High-intensity tumor budding is a strong independent prognostic factor for prediction of lymph node metastasis.

Cytomorphometric Analysis and Assessment of Periodic Acid Schiff Positivity of Exfoliated Cells from Apparently Normal Buccal Mucosa of Type 2 Diabetic Patients

Background: In recent years, important advances have been made in the diagnosis of diabetes mellitus, and new strategies have been put forward for its treatment. The purpose of this study was to analyze the cytomorphometric changes and glycogen content in exfoliated cells of oral mucosa as an adjunct in the diagnosis of diabetes mellitus. Method: The smears were taken from buccal mucosa of 30 type 2 diabetes mellitus patients (study group) and 30 healthy individuals (control group). One smear was stained with rapid Papanicolaou stain and the other with periodic acid Schiff stain (PAS). In Papanicolaou stain smears, the nuclear area, cytoplasmic area and cytoplasmic to nuclear ratio were evaluated from 50 cells in each smear using Image analysis software (Q Win Standard, Leica™) and a research microscope (DM 2500, Leica). PAS-stained smears were analyzed for the presence of glycogen in exfoliated cells. Results: The results showed that the mean nuclear area was significantly higher (p < 0.001) in the study group whereas the mean cytoplasmic area did not exhibit a statistically significant difference (p > 0.001). The mean cytoplasmic to nuclear ratio was significantly lower in the study group (p < 0.001). There was a significant increase in the count of PAS-positive exfoliated cells of the study group as compared with the control group (p < 0.001). Conclusion: The results associated with clinical observations suggest that type 2 diabetes mellitus can produce morphologic and functional alterations in oral epithelial cells, detectable by microscopic and cytomorphometric analysis using exfoliative cytology which can be used in the diagnosis of the disease.

Anthropometric and anthroposcopic analysis of different shapes of faces in group of Indian population: A pilot study

Focus of facial identification research is to isolate features that can be considered a factor of individualization. Anthropometry and Anthroposcopy have been the most important research tools in the field of forensic anthropology. These two methods of observation and data collection are made both on the living and on skeletonized human remains. Anthropometry is a systematic study of human measurements. Morphological analysis of unmeasurable features usually is the area of anthroposcopy. Hence, this study aims to assess different human faces using anthropometry and anthroposcopy. A total of 100 subjects were included in the study, prosopic index given by Martin and Saller was used to classify facial types. An anthropometric measurement was used to assess the facial proportions and anthroposcopy for facial morphology. A significant number of subjects were of hyperleptoprosopic facial type, and with a significant sexual dimorphism observed among the Indian population. Anthroposopic analysis showed common morphological features in each prosopic indices. The findings in our study may throw light on anthropometric and anthroposcopic features of Indians and help significantly in the field of forensic anthropology.