Standardization of honey as a tissue fixative for histopathology: A morphometric study.

Background: Tissue fixation is a crucial step to preserve the tissues in a life-like state with minimal disruption to its cellular and chemical composition for histopathological examination. The search for an effective alternate tissue fixative to the routinely used formaldehyde has gained interest as constant exposure to formaldehyde has proven to be toxic. Honey, an organic substance with high acidity and hygroscopic nature, exhibits tissue fixative properties and has been used in the present study. The present study aimed to standardize honey as a tissue fixative for histopathology by comparing it with formalin. Materials and Methods: In vitro study Oral tissue samples of goat were fixed in 10% honey and 10% formalin solution, respectively, for 24-48 h, followed by routine tissue processing and microscopic examination of 37 slides per group. 2200 epithelial cells (1100 per group) were selected for the computer-aided morphometric image analysis (Fiji-Image J) by three observers. Cell area (CA), cell perimeter (CP), nuclear area (NA), nuclear perimeter (NP), cytoplasmic area (Cyt A), and nuclear-cytoplasmic ratio were the parameters studied. Mann-Whitney U-test (STATA/IC version 16) for inter-group comparison was done and P < 0.05 was considered statistically significant. Results: The probability of epithelial cells in the honey-fixed group to have greater NA, NP, and N/C ratio was about 50%-60%. The probability of epithelial cells in formalin-fixed tissues to have greater CA, CP, and Cyt A was about 70%. Conclusion: Honey is a better nuclear fixative than formalin. Cytoplasmic shrinkage of epithelial cells should be taken into consideration while fixing tissues with honey.

Role of dry ice in decoverslipping of microscopic slides: A new insight.

Background: The process of decoverslipping is often required in a laboratory to review or examine an older slide which tends to fade over time, making it almost impossible to use it for research or study purposes. The sections then need to be re-stained which can only be done after removing the coverslip. The traditional method of decoverslipping using xylene is a time-consuming process. Various methods have been used in the past; however, none were found to be completely effective. Dry ice, the solid form of carbon dioxide, is an easily available, cheap cooling agent with a low freezing temperature (-78.5°C) which was evaluated for its efficacy in decoverslipping process, as an alternative to xylene. Materials and Method: 64 faded haematoxylin and eosin (H&E)-stained histopathology slides were randomly selected and segregated, according to duration of year, into eight major groups. Each group was further divided into four subgroups according to the time that the slides were subjected for decoverslipping. The slides were placed on dry ice and the time was set. Once the coverslip was removed, the slides were placed in xylene to remove any residual mountant. The tissue sections were evaluated for physical disfigurement followed by re-staining with H&E to check for any change in tissue morphology. Result: The mean time taken for removal of coverslip using dry ice was 35 seconds. Conclusion: This technique is easy, fast, and effective, with no tissue loss or compromise in staining quality, thereby preventing xylene toxicity and its effect on the environment.

Spectrum of salivary gland diseases: A 24-year single-institution retrospective study.

Introduction: Salivary glands are exocrine glands and are classified as major and minor salivary glands. Salivary gland pathologies are classified as neoplastic and non-neoplastic. The neoplasms of salivary glands can be benign or malignant. Aim: The aim of the study was to describe the frequency of various salivary gland diseases reported in our institution from 1997 to 2021. Methodology: This was a 24-year retrospective study of salivary gland lesions processed and reported by the Department of Oral Pathology and Microbiology. Data regarding age, gender, site, and diagnosis were obtained and studied. Results: Amongst the total 5928 biopsied cases reported, 6% were salivary gland pathologies. Two hundred sixty-six were non-neoplastic lesions and 81 were neoplastic. The most common non-neoplastic lesion was mucous extravasation cyst. The most common neoplastic lesion was pleomorphic adenoma. Conclusion: The frequency of occurrence of salivary gland lesions in the last 24 years of this institution is almost similar to that stated in other published studies.

Juvenile ossifying fibroma of maxilla: Concurrent presentation of psammomatoid and trabecular variant.

Juvenile Ossifying Fibroma (JOF) is a type of ossifying fibroma which occurs in younger individuals and manifests as trabecular and psammomatoid variants. The nature and behaviour of these variants vary, and they exhibit characteristic histopathological appearance. The solitary presentation of these subtypes is reported in numbers, but co-occurrence of both these entities is very few. Here, we present a case of JOF with the co-occurrence of both trabecular and psammomatoid variants in relation to an incompletely healed extraction socket.

Estimation and comparison of salivary flow rate and its composition in diabetic patients and nondiabetic patients: A pilot study.

BACKGROUND: Diabetes mellitus (DM) is a chronic metabolic disorder characterized by hyperglycemia irregularities in the metabolism of carbohydrates, lipids and protein. It is often associated with the development of microvascular and macrovascular complications and neuropathies. The health of oral tissues is known to be related to the quality and quantity of saliva both of which may be altered in diabetes. AIM: The aim of the present study was to determine the salivary flow rate, electrolytes and total proteins in saliva of Type II diabetic patients. MATERIALS AND METHODS: A total number of 120 participants were included in this study, in which 80 patients were suffering from Type II DM (which included both controlled and uncontrolled diabetes) and 40 nondiabetic persons (controls). The study population included both the genders, with an age range of 40-70 years. The study population was divided into three groups. RESULTS: The values of total protein, sodium, potassium and salivary flow rate among controls, controlled diabetes and uncontrolled diabetes were collected, formulated and multiple comparisons between the groups using the analysis of variance and post hoc Tukey honestly significant difference analysis were done in version 16.0 of SPSS software. CONCLUSION: Studies with larger sample size are warranted to know the exact pathophysiology of controlled and uncontrolled Type II DM in terms of salivary flow rate, salivary electrolytes and total protein.

Adenosquamous carcinoma of the tongue: A case report and an overview of histogenetic concepts.

Adenosquamous carcinoma (ASC), a rare variant of squamous cell carcinoma, is an aggressive and highly infiltrating epithelial neoplasm, which has a distinct histomorphological features comprising simultaneous areas of squamous cell carcinoma and adenocarcinoma. Some researchers consider it as a controversial neoplasm, as it may arise from the ducts of the minor salivary gland or from the overlying surface epithelium. In the head-and-neck region, commonly affected sites include paranasal sinuses, larynx and oral cavity. Areas of occurrence in the oral cavity include palate, tonsillar pillar areas, tongue and floor of the mouth. The prognosis of the neoplasm is considered poor due to its early recurrence, local and distant metastasis and low survival rate. This article reports a case of ASC affecting the posterior part of the tongue in a 50-year-old male patient and an overview of its histogenetic concepts.

Image-Guided Percutaneous Biopsy for Pancreatic Lesions: 10-Year Experience in a Tertiary Cancer Center.

INTRODUCTION: Image-guided biopsy is an important tool in the diagnosis of solid pancreatic masses. This study aims to evaluate the accuracy and complications associated with this procedure. METHODS: We conducted a retrospective cohort study of all patients referred to interventional radiology for pancreatic biopsy at our institution from July 2004-July 2015. Two radiologists independently reviewed the images, technical aspects, and clinical characteristics. RESULTS: A total of 82 patients underwent pancreatic biopsy using a combination of computed tomography (CT) and/or ultrasound guidance. Mean age was 67.8 years with 56% males. Most lesions were located in the head (63.4%), followed by the tail (19.5%), and the body (17.1%). The procedures were performed using the anterior (79.3%), lateral (7.3%), or posterior (13.4%) approach, with direct access in 85.4% of the patients and indirect access (transgastric or transhepatic) in 14.6% of cases. Core biopsies were obtained in 81.7% of cases, and fine needle aspiration was performed in 18.3% of cases. Needle size varied from 18-22 gauge (median 20 gauge). The median number of samples collected was 3 (range 1-8). A conclusive result was obtained in 82.9% of cases. There were 9 (11%) patients with minor complications: minor bleeding, pain, and inadvertent bowel puncture with no clinical repercussion. There were no deaths or major complications. CONCLUSION: Percutaneous image-guided pancreatic mass biopsy can be performed safely with excellent accuracy. Our results are comparable with endoscopic biopsy/aspiration, and percutaneous biopsy may be used as an alternative, particularly at institutions where endoscopy facilities are limited.

Comparison of cheiloscopy, odontometric, and facial index for sex determination in forensic dentistry.

BACKGROUND: The "gender determination" which is an important human identification procedure not only helps in establishing the biological profile from skeletal and dental remains but also in facial reconstruction of unidentified victims. AIM: The aim of this study is to analyze predominant types of lip prints (cheiloscopy), accuracy of mandibular canine index (MCI) (odontometric), and facial index in the study population and to identify whether any correlation among the above parameters could help forensic dentistry in solving crimes. MATERIALS AND METHODS: A pilot study was conducted in 100 individuals, 50 males and 50 females aged between 20 and 25 years. For each individual, the lip prints, MCI, and facial index measurements were recorded on the same day analyzed by two observers. All the analysis was done using SPSS version 14 assessed using t-test and Chi-square test. RESULTS: Type II pattern of lip prints is observed as common pattern among male and female. There is no significant difference in Odontometric analysis. The mean value of facial index analysis in both genders shows highly significant. CONCLUSION: A large-scale study is required in order to validate our results to arrive at definitive results and value.

Evaluation of Salivary Nitric Oxide Levels in Smokers, Tobacco Chewers and Patients with Oral Lichenoid Reactions.

INTRODUCTION: Nitric oxide (NO), a free radical, acts as a signalling molecule affecting numerous physiological and pathological processes. Role of nitric oxide as a mediator in tobacco related habits and the resultant oral lichenoid reactions was assessed. AIM: The aim of the study is to evaluate and compare the salivary nitric oxide levels in normal patients with that of smokers, tobacco chewers and patients with oral lichenoid reactions. MATERIALS AND METHODS: One hundred and twenty patients were enrolled in the study which included 30 healthy patients without any chronic inflammatory lesion and habit as controls (group I), 30 smokers without the habit of tobacco/betel nut chewing and any oral lesion (group II), 30 tobacco chewers without the habit of smoking and any oral lesion (group III) and 30 histologically confirmed cases of oral lichenoid reaction with the habit of tobacco usage (group IV). Saliva from these patients was collected and the nitrite concentration was assessed. RESULTS: Our results concluded that there was highly significant increase in the nitric oxide levels in smokers, tobacco chewers and patients with oral lichenoid reactions compared to that of controls. Also, there was a significant increase in nitric oxide levels in patients with smoking associated oral lichenoid reactions in comparison with smokers and in patients with lichenoid reactions associated with tobacco chewing in comparison with tobacco chewers. CONCLUSION: Estimation of salivary nitric oxide levels is a simple, non-invasive procedure and could be analysed to suggest the role of nitric oxide in the pathogenesis of these lesions. The increased activity of the enzyme may indicate that nitric oxide has a pathophysiological role in these lesions.

Giant cell fibroma: A clinicopathological study.

OBJECTIVE: Giant cell fibromas (GCF) of the oral cavity are found predominantly in Caucasians and rarely in other races. This retrospective study was done to evaluate the clinicopathological features of GCFs in a sample of Indian population. MATERIALS AND METHODS: 21 oral GCF cases were investigated from the year 1995 to 2010. Clinical data and microscopic features were reviewed and analyzed. RESULTS: The mean age of patients at the time of diagnosis was 39years. Oral GCF occurred in patients between 6 and 67 years of age. The lesions were 4-17 mm in greatest dimension. GCF frequently has the provisional diagnosis of fibroma or papilloma. All tumors were treated by total surgical excision and no recurrence was reported. The consistent and diagnostic feature was the presence of large stellate giant cells, usually with one or two nuclei. Multinucleated giant cells were seen occasionally. These giant cells were most numerous in the connective tissue beneath the epithelium. CONCLUSION: Though there are distinct histopathologic features for GCF, its clinical presentation and prognosis are similar to the conventional fibroma/fibroepithelial polyp.

Plasma cell granuloma of lip.

Plasma cells are medium-sized round-to-oval cells with eccentrically placed nuclei, usually found in the red pulp of the spleen, tonsils, medulla of the lymph nodes, nasal mucosa, upper airway, lamina propria of the gastrointestinal tract, and sites of inflammation. Plasma cell granuloma is a rare reactive tumor-like proliferation composed chiefly of plasmacytic infiltrate. Here, we present a case of plasma cell granuloma of lip in a female patient.

Evaluation of oral submucous fibrosis using ultrasonographic technique: a new diagnostic tool.

AIM: The aim of this study is to evaluate oral submucous fibrosis (OSMF) by clinical and histopathological examination, and compare the results with those from ultrasonographic technique. MATERIALS AND METHODS: 30 clinically diagnosed OSMF patients were subjected to both ultrasonographic and histopathological evaluation before treatment. Later, only ultrasonographical examination was done during 4 th and 8 th week of treatment. Prognosis of the lesion for the treatment was evaluated. Peak systolic velocity (PSV) of blood in the lesional area was statistically analyzed. 10 normal individuals without any mucosal lesions were considered as the control group. RESULTS: In normal individuals, ultrasonography delineates normal mucosa with uniform fine mottled appearance with interspersed hypoechoic areas. Color Doppler and spectral Doppler depicts uniform distribution of blood vessels and normal peak systolic velocity of blood respectively. All OSMF patients were diagnosed upon clinical and histopathological examination. Clinical examination revealed 14 individuals with unilateral palpable fibrotic bands and 16 individuals with bilateral fibrotic bands whereas, ultrasonographic evaluation revealed 6 individuals with unilateral fibrotic bands and 24 individuals were with bilateral fibrotic bands, which was statistically significant. Ultrasonography demonstrated number, length and thickness of the fibrotic bands. Color Doppler and spectral Doppler showed decreased vascularity and PSV in lesional area. Prognosis evaluation revealed 25 cases of good prognosis and 5 cases were showed poor prognosis. Wilcoxon Signed Ranks Test revealed no significant difference of PSV was seen in poor prognosis patients. CONCLUSION: Ultrasonography is a non-invasive diagnostic tool for OSMF. It could be a better diagnostic tool compared to clinical and histopathological examination.

Immunohistochemical evaluation of mast cells and vascular endothelial proliferation in oral submucous fibrosis.

INTRODUCTION: Oral submucous fibrosis (OSMF) is a chronic, progressive, scarring disease that predominantly affects the people of south-east Asian origin. AIM: The present study is aimed at comparing and correlating the mast cell density (MCD) and micro vascular density (MVD) in normal mucosa and different grades of OSMF and to analyze their role in the disease progression. MATERIALS AND METHODS: MCD was assessed immunohistochemically using anti mast cell tryptase and MVD was assessed using anti-factor VIII related von Willebrand factor. RESULTS: The one way comparison of MVD and MCD in normal mucosa and among different grades of OSMF showed a significant increase in MCD and MVD among OSMF cases. Correlation analysis using Pearson correlation coefficient, showed positive correlation between MCD and MVD i.e. as MCD increases there is an exponential increase in MVD. CONCLUSION: The increase in MVD and MCD reveals their role in the pathogenesis of OSMF, a lesion characterized by progressive fibrosis in early stages and failure of degradation or remodeling in the advanced stages.

Measurement of blood E2F3 mRNA in prostate cancer by quantitative RT-PCR: a preliminary study.

The use of serum prostate-specific antigen (PSA) measurements necessitates biopsies for accurate prostate cancer (CaP) diagnosis. Overall efficiency of accurate diagnosis, when PSA levels are used alone, is less than 60%. E2F3 was evaluated as an alternative biomarker using patient blood samples. Expression levels were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and correlated with accurate clinicopathological data. Statistical analysis demonstrated significant differences in E2F3 expression levels (p<0.0001), and high levels of discrimination (receiver operator curve/area under curve analysis values (AUC) >0.88), in particular at early stages of disease development, between benign disease and localized CaP. Limited levels of discrimination were observed at the later stages of disease development, between localized and metastatic disease (p=0.076, AUC=0.633). A cut-off point of 0.34 with high specificity for benign disease (92.3%) and sensitivity for CaP diagnosis (81.0%) was identified. At this cut-off point, 85% patients were correctly diagnosed with either malignant or benign disease. This study demonstrates the strength of E2F3 as a potential marker for discriminating benign and malignant disease, addressing the current limitations of serum PSA measurements.

An unusual case of stroke in a young patient.

The patent foramen ovale (PFO) has been increasingly implicated in the aetiology of stroke, particularly in young patients with no other identifiable cause (cryptogenic stroke). A case of Parinaud's syndrome secondary to thromboembolism from a PFO is reported here.