Assessing the Quality of Long-Term Stored Tissues in Formalin and in Paraffin-Embedded Blocks for Histopathological Analysis.

Introduction: Formalin is the most commonly used fixative which enables for long-term storage of specimens and preserves morphologic features allowing the microscopic evaluation for future research analysis. Archival collections of the tissue serve as a reliable tool for diagnostic research purpose. They have an important role in on-going patient care, allows for evaluation of recurrent cases for diagnostic purpose and rare case specimens can also be used as an educational tool as well as for further biomedical research purposes. However, studies assessing quality and their usefulness for such purposes are scanty. Hence, the present study is aimed at evaluating and comparing the tissue changes after long-term storage in formalin as well as in paraffin-embedded blocks. Methodology: Three study groups include specimens stored in formalin for a minimum of 5 years (long-term fixed tissue) and their corresponding paraffin-embedded old tissue blocks along with freshly fixed tissues taken as controls which were subjected to routine histopathological procedures and were assessed for macroscopic and microscopic evaluation.Chi-square test and Z-proportion tests were considered for statistical analysis. Results: Prolonged storage of the tissues in formalin showed variation in color and consistency, difficulty in cutting during grossing with inadequate sectioning characters, loss of tissue integrity and architecture, and inadequate nuclear and cytoplasmic details. Conclusion: On histological analysis, prolonged formalin-stored specimens showed deleterious effects than archival blocks. Hence, it can be proposed that tissues are better preserved in paraffin blocks rather than in formalin for further biomedical research purposes.

Age Estimation with Cemental Annulation Using Light, Phase Contrast and Polarized Microscopy.

INTRODUCTION: In forensic science, the determination of age plays a vital role in the identification of bodies and persons associated with crimes. Teeth are frequently better conserved than any other human remains, so their use for identifying age at death is vital. The root portion of the teeth is covered by a thin calcified layer called cementum, the annulations of which is considered to be helpful in age estimation. The objective of the study was to ascertain and compare the accuracy and efficiency of age estimation between light, polarized, and phase-contrast microscopy in the ground and decalcified sections of the tooth stained with hematoxylin and eosin by light microscopy and picrosirius red (PSR) by polarized microscopy. MATERIALS AND METHODS: Fifty extracted teeth were collected and stored in a solution of 10% formalin. The middle one-third portion of the root was used, later sectioned into two halves using carborundum disc. One-half of it was used to prepare ground sections, which was studied with light, polarized, and phase-contrast microscopy. The other half was decalcified with 10% formic acid, processed, and two sections of 5 -µm thickness were prepared. One was stained with PSR stain and the second section was stained with hematoxylin and eosin (H and E) stain, One was stained with PSR stain and the second section was stained with hematoxylin and eosin (H and E) stain. Statistical analysis was performed using Z-test and Karl Pearson's correlation coefficient. RESULTS: No statistically significant difference was observed between actual and calculated age in the ground sections, while there was a statistically significant difference observed between actual and calculated age in decalcified sections stained with H and E and PSR. A strong positive correlation was observed between actual and calculated age by Karl Pearson correlation coefficient test. CONCLUSION: Cemental annulation and phase contrast microscopy can be reliably utilized in forensic science to establishing age, especially among young and middle age group individuals.

Comparison of staining adequacy between tissues stored in formalin and paraffin embedded blocks for prolonged duration.

INTRODUCTION: Tissues submitted for histopathological diagnoses are routinely fixed in formaldehyde fixative which facilitates tissue storage for the extended period of time for further investigation or research purpose. The literature has reliable data relating to immunohistochemistry and molecular studies done using tissue fixed in formalin for longer duration and in paraffin-embedded tissue blocks. We have attempted to evaluate staining adequacy of archival specimens using hematoxylin and eosin (H&E) and trichrome stains. METHODOLOGY: We compared staining adequacy in tissues stored in formalin for a minimum period of 5 years labeled as long term fixed tissues (LFT) and corresponding 5-year-old paraffin-embedded tissue blocks of same cases available in the archives designated as old tissue blocks (OTB), along with freshly fixed tissues (FFT) used as controls. Fifty-one tissue sections in each study group were stained with H&E, Mallory's trichrome and Van Giesons' and microscopic evaluation was carried out using Research Microscope B × 51. RESULTS: Staining adequacy of the tissue sections was found to be superior in FFT, followed by OTB and LFT using all three stains. Staining for H&E, was found to be superior as compared to trichrome stains. CONCLUSION: Trichrome stains were poorly demonstrated in LFT indicating the possible alteration in protein structure and tissue architecture in LFT whereas H&E, stained LFT specimens showed relatively good staining character. Hence, OTB proved to be better as compared to LFT in terms of staining quality and hence archiving tissue in paraffin embedded tissue blocks serve as valuable bio-bank and source for prospective studies.

Sexual dimorphism of enamel area, coronal dentin area, bicervical diameter and dentinoenamel junction scallop area in longitudinal ground section.

INTRODUCTION: Sex determination plays an important role in forensics; several studies done using radiographs, model cast and animal teeth have shown that males possess larger tooth crown and more dentin than that of the females. As physical sectioning of tooth provides more accurate measurement of enamel thickness when compared with other methods, the present study was done to evaluate and compare enamel area (EA), coronal dentin area (CDA), bi-cervical diameter (BCD), average enamel thickness (AET) and dentinoenamel junction scallop area (DEJ-SA) in longitudinal ground sections of first premolars between males and females. MATERIALS AND METHODS: A total of 60 extracted first premolar teeth were used for the study, of which 30 were from male and 30 were from female. A longitudinal ground section of 15 maxillary and 15 mandibular premolars of approximately 50 µm was prepared buccolingually at the center of each tooth and mounted on the slide. Multiple photomicrographs were captured with the help of Image Analysis System Software-Progres, Speed XT core 3. EA, CDA, length of the dentinoenamel junction and BCD were measured. Average DEJ-SA and AET were also calculated. RESULTS: Mann-Whitney U- test was used for statistical analysis. It was found that EA and AET were significantly greater in females than in males. On the contrary, it was found that CDA was significantly greater in males than in females. However, no significant difference was found in BCD and DEJ-SA between males and females. CONCLUSION: Permanent first premolar can be reliably used in the field of forensic in establishing gender of individuals by measuring its EA, CDA and AET.

Chronic Recurrent Non-specific Parotitis: A Case Report and Review.

BACKGROUND: Chronic recurrent non-specific parotitis is characterized by recurrent episodes of swelling and pain of unknown etiology in the parotid gland. Sialography is a hallmark in the diagnosis of salivary gland disorders; newer imaging modalities like CT-Sialography, sialoendoscopy and MRI can be used. Various treatment modalities have been tried, from conservative approach to surgical excision depending on the recurrence rate and severity of the condition. Although symptomatic treatment with antibiotics and analgesic, injection of intraductal medicament, aggressive treatment like duct ligation or excision of gland are some of the treatment modalities, there is no established algorithm as to which treatment method should be opted in such clinical situation. CASE DETAIL: A 20 years old male patient reported with pain and salty taste in the mouth that had began before a week. Examination revealed an elevated right parotid papilla; ropy, cloudy appearing saliva was oozing out on milking the gland. Unstimulated and stimulated whole salivary flow rate was assessed using drooling method. Sialography was used as a diagnostic and a therapeutic aid. In our case, sialography as a treatment showed a good response with no recurrence after two years of follow-up. We highlighted the role of sialography as a therapeutic aid. CONCLUSION: Recurrent attacks significantly affect the quality of life and also lead to progressive gland destruction. Preventing or reducing the frequency of recurrence remains the goal of therapeutic procedure. Hence, conventional sialography is useful in the diagnosis and also effective as a therapeutic aid in recurrent parotitis.

Picrosirius Red and Polarization Microscopy - A Tool for Gender Differentiation.

INTRODUCTION: Forensic dentistry is a branch of dentistry which in collaboration with legal profession serves an important role to maintain justice system of a country. Forensic dentists play a major role in identification of an individual. Within the literature various methods have been found to be useful in gender differentiation. An attempt was made for differentiation of gender using picrosirius red and polarization microscopy. AIM: To evaluate picrosirius red and polarization microscopy as a tool for gender differentiation by observing birefringence pattern and distribution of thick and thin collagen fibers in males and females. MATERIALS AND METHODS: Labial mucosal tissue obtained from 30 deceased individuals (18 male and 12 female) during autopsy was fixed in 10% formalin at 12th hour. Tissue was processed, sectioned and stained using picrosirius red stain and the birefringence pattern of collagen fibers were studied with polarization microscope. The results were statistically analyzed using Z-test and one-way ANOVA to draw the significance. RESULTS: The proportion of thick and thin fibres among males and females were compared. It was found that there was statistically significant difference in proportion of thick and thin fibers between male and female. Thick fibres in males were (78.13%) more than females (65.74%) and thin fibres were more in females (34.24%) than males (21.32%). CONCLUSION: Picrosirius red and polarization microscopy may be used as a tool for gender differentiation. Yet the manner of death has to be considered during gender differentiation using this method, as in the present study out of 30 cases studied three cases of death due to debilitating diseases and poison consumption showed altered collagen distribution.

Study of Collagen Birefringence in Different Grades of Oral Squamous Cell Carcinoma Using Picrosirius Red and Polarized Light Microscopy.

Objectives. The present study was done to evaluate birefringence pattern of collagen fibres in different grades of oral squamous cell carcinoma using Picrosirius red stain and polarization microscopy and to determine if there is a change in collagen fibres between different grades of oral squamous cell carcinoma. Materials and Methods. Picrosirius red stained 5 µm thick sections of previously diagnosed different grades of squamous cell carcinoma and normal oral mucosa were studied under polarization microscopy for arrangement as well as birefringence of collagen fibres around tumour islands. Results. It was found that thin collagen fibres increased and thick collagen fibres decreased with dedifferentiation of OSCC (P < 0.0001). It was observed that there was change in polarization colours of thick fibres from yellowish orange to greenish yellow with dedifferentiation of OSCC indicating loosely packed fibres (P < 0.0001). Conclusion. There was a gradual change of birefringence of collagen from yellowish orange to greenish yellow from well to poorly differentiated squamous cell carcinoma, indicating that there is a change from mature form of collagen to immature form as tumour progresses. Studying collagen fibres with Picrosirius red for stromal changes around tumour islands along with routine staining may help in predicting the prognosis of tumour.

Association of root caries with oral habits in older individuals attending a rural health centre of a dental hospital in India.

BACKGROUND: Many risk factors can compromise an older adult's systemic health. Among the oral ailments in the elderly, root caries is a significant one which causes tooth loss in them. Hence, there is a need to have a baseline data for understanding problem of root caries in elderly population and factors which affect its prevalence. AIMS: a)To asses the prevalence of root caries in older individuals in a rural health centre in India. b) To asses the relationship of oral habits with root caries. MATERIALS AND METHODS: The study included 210 elderly dentate and consenting individuals (123-females, 87-males) aged 55 to 75 y and above. Demographic and health behaviour data were collected through personal interviews. The subjects were examined for root caries. Statistical analyses of the data were done using chi-square and multiple logistic regression analysis. RESULTS: Out of all, 94.76% of elderly patients examined had gingival recession in one or more teeth. The prevalence of root caries was 41.9%. The prevalence of root caries was significantly associated with age, perceived dryness of mouth, smoking, smoking and tobacco chewing and tobacco chewing only (p<0.05). There was significantly higher root caries in the age group of 75 years and above (OR-3.67). CONCLUSION: It was evident from our study that root caries prevalence was high in elderly population. Age, root surfaces with recession, deleterious oral habits such as smoking, tobacco chewing, and dryness of mouth had a definite effect on the prevalence of root caries.

Diabetes and Oral Health: A Case-control Study.

BACKGROUND: Diabetes mellitus, according to World Health Organization (WHO) is a silent epidemic which affects large number of people around the world and is directly related to the oral health status of the patients. OBJECTIVES: To know the prevalence of common dental diseases such as dental caries, periodontal diseases (pyorrhea), and treatment needs in a group of adult diabetic patients in private medical establishments of Tumkur city, south India, in comparison with non-diabetic patients. To create awareness among general medical practitioners about the common oral manifestations of diabetes and the importance of periodical dental check up for diabetics. METHODS: A group of 300 diabetic patients (males = 186, females = 114) and a control group of 300 non-diabetics (males = 180, females = 120) matched by age and sex were examined according to WHO criteria, for a period of eight months. RESULTS: The prevalence of dental caries was comparatively more in non-diabetics (32.3%) than in diabetics (13.6%). However, the prevalence of periodontal diseases (pyorrhea) was more in diabetics (92.6%) when compared to non-diabetics (83%). CONCLUSIONS: Oral health is an integral part of general health. Though dental caries was comparatively low in diabetics, periodontal status was compromised. Complex treatment needs was more in the diabetics (58%) when compared to controls (41%). Regular follow-up of dental problems of the diabetics and oral health education is much required.