Clinical and histopathological correlation of oral malignancy and potentially malignant disorders based on a screening program at high-risk population in Tamil Nadu, India.

Background: There is a high incidence of oral cancer and oral potential malignant disorder observed in southeast Asian countries such as India. Our study aimed to assess the correlation between screening and histopathological diagnosis and to predict the specificity and sensitivity of chair-side/field-based assessment of the oral lesion. Materials and methods: A total of 40,852 subjects aged between 20 and 60 years were screened in the 1st phase of the study, suspected lesions were stained with toluidine blue (Manufactured by Otto Chemicals private limited, India) at two time points, those who stained positively during the two points were taken up for biopsy. Provisional diagnosis was later correlated with histopathological diagnosis. Results: Subjects who underwent biopsy had a mean age of (49.01 ± 9.8 years), Leukoplakia (1.5%) was the most common lesion observed among tobacco users, interestingly it had the least correlation (39.6%) in diagnosis, Overall sensitivity (88%) and a positive predictive value (80%) was high for clinical diagnosis of OPMD in our study. Conclusion: Correlation of clinical and histopathological diagnosis observed in our study confirms higher yield of true positives while screening in remote and vulnerable populations, which would assure a better quality of life for these subjects.

Povidone iodine, hydrogen peroxide and chlorhexidine mouthwashes reduce SARS-CoV2 burden in whole mouth fluid and respiratory droplets

SARS-CoV2 is transmitted primarily through oral mouth secretions and respiratory droplets. Commercial mouthwashes, povidone iodine (PI), hydrogen peroxide (HP) and chlorhexidine (CHX) have been tested in cell culture and RT-PCR studies for their efficacy to reduce SARS-CoV2 burden. Here, we evaluated SARS-CoV2 burden in whole mouth fluid (WMF) and respiratory droplets (RD) samples before and after the use of PI, HP or CHX mouthwashes in hospitalized COVID-19 patients using RT-PCR and rapid antigen test (RAT). Thirty-six SARS-CoV2 RT-PCR-positive in-patients were randomly assigned to one of the four groups: 20 and 60 minutes after 1% w/v PI or 1.5% HP; 90 and 180 minutes after 1.5% HP or 0.2% w/v CHX. WMF and RD samples were collected concurrently at baseline and after the two different time points. RD (92%) showed a higher reduction in SARS-CoV2 burden than WMF samples (50%; p=0.008). SARS-CoV2 burden was statistically lower at both 20 minutes (p=0.02) and 60 minutes (p=0.03) with PI; at 20 minutes with HP (p=0.0001); and 90 minutes with CHX (p=0.04). The overall and individual mean logarithmic reductions in the WMF and RD samples were greater than 1.0 at 20, 60 and 90 minutes after PI, HP or CHX. RAT-positive patients at 90 minutes post-treatment (n=3) demonstrated a one log increase in virus copies. Among the three RAT-negative post-treatment patients, SARS-CoV2 burden declined by one log in two while the third patient had a slight increase in RNA copies. In conclusion, we have shown for the first time that the mouthwashes, PI, HP and CHX can reduce the SARS-CoV2 burden in the concurrently collected RD and WMF samples. RAT is more appropriate than RT-PCR to evaluate the efficacy of the mouthwashes.

Implications in the quantitation of SARS-CoV2 copies in concurrent nasopharyngeal swabs, whole mouth fluid and respiratory droplets

ImportanceThe nasopharyngeal swab (NPS) is considered the ideal diagnostic specimen for Covid-19, while WMF is recently promoted due to collection simplicity and importance in disease transmission. There is limited knowledge on the relative viral load in these samples - NPS, whole mouth fluid (WMF) and respiratory droplets (RD; another important source in transmission), on how the loads vary with disease severity and on how much virus is shed. ObjectiveTo quantify and compare SARS-CoV2 copies in the NPS, WMF and RD samples, and correlate with disease severity. DesignCross sectional study. SettingTertiary care multi-speciality hospital with limited resources in a low-to-middle income country. ParticipantsEighty suspected COVID-19 patients were recruited from the COVID-19 out-patient clinic and hospital isolation wards. InterventionConcurrent NPS, WMF and RD samples were collected from all the recruited patients and tested for SARS-CoV2 copies by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Main outcomes and measuresThe main outcome was COVID-19 measured by SARS-CoV2 quantitative RT-PCR in NPS samples. COVID-19 disease severity was determined according to NIH criteria. Virus shedding was defined as the presence of SARS-CoV2 copies in the WMF and RD samples. ResultsSARS-CoV2 was detected in 55/80 (69%) of the NPS samples. Of these 55, WMF and RD samples were positive in 44 (80%) and 17 (31%), respectively. The concordance of WMF with NPS was 84% (p=0.02). SARS-CoV2 copy numbers were comparable in the NPS (median: 8.74x10^5) and WMF (median: 3.07x10^4), but lower in RD samples (median: 3.60x10^2). Patients with mild disease had higher copies in the NPS (median: 3.46x10^6), while patients with severe disease had higher copies in the WMF (median: 1.34x10^6) and RD samples (median: 4.29x10^4). The 25-75% interquartile range of NPS SARS-CoV2 copies was significantly higher in the WMF (p=0.0001) and RD (p=0.01) positive patients. Conclusion and relevanceSARS-CoV2 copies are highest in NPS samples. WMF is a reliable surrogate sample for diagnosis. High copy numbers in the NPS imply initial virological phase and higher risk of virus shedding via WMF and RD. Key pointsO_ST_ABSQuestionC_ST_ABSHow the numbers of SARS-CoV2 copies in nasopharyngeal swab (NPS) samples might reflectvirus shedding from the whole upper aerodigestive tract and indicatedisease severity? FindingsIn this cross-sectional study involving 80 suspected COVID-19 patients, the data indicate higher SARS-CoV2 copies in NPS samples of patients with mild disease,and in the whole mouth fluid (WMF) and respiratory droplet (RD) samples of patients with severe disease. Patients with higher SARS-CoV2 copies in the NPS shed the virus in the WMF and RD samples at statistically higher levels. MeaningHigh SARS-CoV2 copies in NPS samples imply initial virological phase withhigh levels of shedding through both WMF and RD.

Detection of SARS-CoV2 antigen in human saliva may be a reliable tool for large scale screening

IntroductionSARS-CoV2, the aetiological agent of the current COVID-19 pandemic, has been detected in saliva and recently implicated in several oral diseases. Collection of nasopharyngeal swabs (NPS) and detection by reverse transcriptase-polymerase chain reaction (RT-PCR) requires medical / technical expertise. A reliable and easy to handle point-of-care (POC) test is highly desirable, especially to curb transmission. Therefore, in this study, we evaluated a commercially available POC rapid antigen test (RAT) for the detection of SARS-CoV2 antigens in the saliva of RT-PCR confirmed positive and negative patients. MethodsThirty saliva samples of 10 saliva RT-PCR negative and 20 saliva RT-PCR positive patients were tested by RAT. ResultsRAT was negative in 10/10 (100%) RT-PCR-negative samples; positive in 9/20 (45%) RT-PCR-positive samples; concordance was 63% (p=0.001). Patients with positive RAT had higher virus copies in their NPS samples compared to the RAT-negative patients. This difference was also statistically significant (p=0.01). ConclusionThus, the POC RAT may be used to detect SARS-CoV2 as a reliable tool for self-testing, large-scale population screening and emergency medical/dental screening. Patients negative by RAT should be confirmed by RT-PCR.

Evaluating and comparing the morphological and histopathological changes induced by erbium:yttrium-aluminum-garnet laser and diamond bur on enamel, dentin and pulp tissue.

AIM: Lasers are used for different types of dental treatments. Using the erbium:yttrium-aluminum-garnet (Er:YAG) laser to remove dental hard tissue is simple, advantageous and influences the type of cavity preparation, whether conventional or conservative in nature. The aim of the present study was to evaluate and compare the morphological and histopathological changes in the enamel, dentin and pulp tissue of the teeth treated by Er:YAG laser and conventional burs. METHODS: A conventional class I cavity was prepared in orthodontic patients by laser and bur. The teeth were extracted and analyzed for morphological changes using a scanning electron microscope, ground sections and histopathological changes under a light microscope. RESULTS: The time with laser was longer than the conventional methods. The lased cavity showed irregular appearance with absence of smear layer which is suitable for the resin restoration. The ground section and the histopathological study showed no differences between the groups. CONCLUSION: The Er:YAG laser is effective in the removal of dental hard tissue without damaging the pulp when coupled with ideal energy output. It is widely used in different dental fields. It needs time to be accepted by dentist and patients and further studies are required to explore its advantages.

Identification of Helicobacter pylori in Saliva of Patients with and without Gastritis by Polymerase Chain Reaction.

AIMS AND OBJECTIVE: The aim of this study was to identify the presence of Helicobacter pylori in saliva of patients with and without gastritis by polymerase chain reaction (PCR) method. MATERIALS AND METHODS: The study comprised 20 patients in Group I presenting with various symptoms of gastritis and 10 asymptomatic subjects in Group II. The intestinal endoscopy antral biopsies were collected from 20 symptomatic patients with gastroduodenal disorders. The saliva specimens were taken from all patients before endoscopy. PCR was performed using genomic DNA, isolated from the saliva and the biopsies of the patients as the template to detect the presence of the 16S ribosomal RNA gene in H. pylori. RESULTS: In Group I, 10 (50%) cases of clinical gastritis were positive for H. pylori by endoscopy biopsy and 10 (50%) were negative. Of the 10 endoscopy biopsy positive cases for H. pylori, eight were PCR positive in saliva and two were negative. Of the 10 endoscopy biopsy negative cases, three were PCR positive for H. pylori in saliva and seven were negative. In Groups II, four were symptomatic for gastritis and six were negative. Of the six gastritis negative cases, three were PCR positive, four were gastritis positive, and three were PCR positive. Sensitivity and specificity of PCR were found to be 80% and 70%, respectively. The positive predictive and negative predictive values of PCR in saliva were 72.7% and 77.7%, respectively. CONCLUSION: PCR analysis of saliva may be handy in identification of H. pylori and serves as a noninvasive technique to diagnose and monitor the prognosis.

Interference RNA in immune-mediated oral diseases - minireview.

Immune-mediated oral disorders are characterised by their chronicity, and some are refractory to treatment. Interference RNA (iRNA) has been implicated in the underlying mechanism of such immune-mediate oral and refractory inflammatory oral diseases. iRNA-based understanding of the mechanism in these diseases may help to produce non-invasive diagnostic methodologies and treatment modalities of such drug non-responsive diseases. Oral lesions in these immune-mediated diseases can precede the occurrence of lesions in other regions of the body. The early diagnosis and treatment of these drug non-responsive diseases might benefit the patient by reducing chronicity and probably even resolving the disease. This aim of the present minireview is to give an overview of the possible implications of iRNA on the pathogenesis, diagnosis, and treatments of immune-mediated and inflammatory oral diseases. The manuscript can form the framework for research on iRNA in these immune-mediated oral disorders.

Comparative study on the microbiological features of angular cheilitis in HIV seropositive and HIV seronegative patients from South India.

OBJECTIVE: This study was designed to compare the microbiological features of angular cheilitis (AC) in human immunodeficiency virus (HIV) seropositive and HIV seronegative individuals, in a group of south Indians. MATERIALS AND METHODS: Swabs from oral commissures of 46 patients were obtained and inoculated on to Sabouraud's dextrose agar (SDA) supplemented with chloramphenicol, blood agar (BA) and MacConkey's agar (MCA) plates and cultured. α-hemolytic Streptococci, Staphylococcus albus, Staphylococcus aureus, Candida species, Klebsiella species and Pseudomonas species were cultured. Candidal colonies were further speciated by the conventional biotyping technique. RESULTS: In AC of HIV seropositive patients Candida albicans and Staphylococcus aureus were more prevalent than that in HIV seronegative patients. Incidentally in patients with CD4 cell count less than 200 there was an increase in the incidence of Candidal and Staphylococcus aureus colonization when compared to patients with CD4 cell count higher than 200. CONCLUSION: The present study suggests a definite difference in the microbial flora of AC in HIV seropositive patients than that of HIV seronegative population.

Assessment of oral mucosa in normal, precancer and cancer using chemiluminescent illumination, toluidine blue supravital staining and oral exfoliative cytology.

CONTEXT: Carcinoma in an early stage of development is hard to detect clinically because the lesion may not be palpable and color of the lesional tissue is not necessarily different from the color of the surrounding mucosa. In order to improve the efficacy of the diagnosis, techniques are being developed to complement clinical examination and to facilitate the identification of initial carcinomas. AIMS: To find out the efficacy of chemiluminescent illumination (ViziLite™) for the diagnosis in precancer and cancer patients and compare this result to toluidine blue staining and oral exfoliative cytology. MATERIALS AND METHODS: This study was done in 3 groups. Each group consists of 10 cases. Group I consists of normal appearing mucosa. Group II and III consist of clinically diagnosed pre-cancer and clinically suggestive of cancer respectively. Chemiluminescent illumination, toluidine blue supravital staining, oral exfoliative cytology and biopsy were performed in all cases. STATISTICAL ANALYSIS USED: SPSS version 10.05 was used to calculate positive and negative predictive values. RESULTS: In Group I, all 10 patients showed negative result to ViziLite™. 8 patients showed positivity and 2 patients showed negativity to ViziLite™ test in Group II. 9 patients were positive and one patient was negative for ViziLite™. CONCLUSIONS: Chemiluminescent illumination test was sensitive for precancerous and cancerous lesions, which presented as keratotic lesions and red-white lesions. It showed negative result to erosive lesions. Toluidine blue staining test was reliable in precancerous and cancerous lesions, which present as erosive and red-white lesions. It showed negative result to keratotic lesions. Oral exfoliative cytology has diagnostic value in cancer patients than in precancer patients. These Results indicate that chemiluminescent illumination test is relatively reliable and accurate than toluidine blue staining test and useful chair side diagnostic test.

Immunohistochemical detection of human telomerase reverse transcriptase in oral cancer and pre-cancer.

PURPOSE: Telomerase is a specialized ribonucleoprotein complex that stabilizes telomeres by adding "TAG" repeats to the end of chromosomes. The catalytic subunit of telomerase is human telomerase reverse transcriptase (hTERT), whose expression is the critical determinant of telomerase activity. Telomeres and telomerases play an important role in the longevity of cell and are known to conform "immortalization" on neoplastic cells. Although there exists a lot of information on telomerase in oral cancer, very little is known about their expression in leukoplakia and oral submucous fibrosis (OSF). This study addresses this lacuna. MATERIALS AND METHODS: In this preliminary study, immunohistochemistry (IHC) was used to detect the expression of hTERT protein in oral squamous cell carcinoma (OSCC) (n=30), leukoplakia (n=15), OSF (n=15) and normal oral mucosa (n=10). The cellular localization of immunostain, intensity of stain, mean nuclear labeling index (LI) and mean nuclear labeling score (LS) of hTERT protein were studied. A total number of 1000 cells were counted in each slide. All the data were analyzed using SPSS software version 10.0.2. The cellular localization of cytoplasmic/nuclear/both of hTERT stain, staining intensity and LI were compared across the groups using Pearson's χ2 test. The mean LI and LS for OSF, leukoplakia, OSCC and normal were compared using analysis of variance (ANOVA). A P-value <0.05 was considered to be statistically significant. RESULTS: The mean nuclear LI increased from OSF (22.46±4.53), through normal (28.3±12.3) to OSCC (47.56±21.30) (P=0.002) and from normal (28.3±12.3), through leukoplakia (44.06±14.6), to OSCC (47.56±21.30) (P=0.00). The mean nuclear labeling score was observed to increase from OSF (37.8±15), through normal (64.9±30.7), to OSCC samples (106.9±29.77) (P=0.00) and from normal (64.9±30.7), through leukoplakia (85.6±25.1) to OSCC samples (106.9±29.77) (P=0.00). CONCLUSION: There was increased expression of hTERT protein in OSCC and leukoplakia samples when compared to normal oral mucosa. The cellular localization, LI and LS in OSF were significantly different from OSCC and leukoplakia.

Salivary glucose levels and oral candidal carriage in type II diabetics.

OBJECTIVE: The aim of this study was to detect salivary glucose levels in diabetic and nondiabetic subjects, to study the relationship between salivary glucose levels and salivary candidal carriage, and to determine if salivary glucose levels could be used as a noninvasive tool to monitor glycemic control in diabetics. STUDY DESIGN: A total of 150 adults, 100 with type 2 diabetes and 50 without diabetes (control subjects), aged 40-60 years, participated in the study. Diabetic status was determined by estimation of random nonfasting plasma glucose levels and glycosylated hemoglobin levels. Both unstimulated and stimulated saliva were collected and investigated for glucose levels and colony-forming units (CFU) of Candida. Salivary glucose levels were measured using the glucose-oxidase method. RESULTS: Salivary glucose levels were significantly higher in diabetics than nondiabetics. There was a significant positive correlation between salivary and plasma glucose levels. Candidal CFUs were significantly higher in diabetic subjects and showed a significant positive correlation with salivary (unstimulated and stimulated) glucose levels. CONCLUSIONS: These results show that salivary glucose concentration is a potentially useful noninvasive tool to monitor glycemic control in diabetic patients. Increased salivary glucose is associated with increased prevalence of oral Candida in these subjects.

Oral Candida species in healthy and HIV-infected subjects in Chennai, South India

Objective: Candidiasis is the most common fungal infection in human immunodeficiency virus (HIV) - infected individuals. As there is sparse data on the oral <I>Candida</I> species in HIV- infected individuals in India, we characterized <I>Candida</I> species from the oral cavity in two cohorts - with and without HIV infection and with presence or absence of clinical oral candidiasis, in Chennai, South India.<br>Methods: Saliva samples were collected from 147 consecutive study participants by the oral rinse technique. <I>Candidal</I> species were isolated by culturing specimens on Sabouraud‘s dextrose agar. The pure cultures so derived were speciated using the commercially available ID32C system, and the results were interpreted using APILAB plus software.<br>Results: In the HIV seropositive group, the most commonly isolated candida species was <I>C.albicans</I> (86%) followed by <I>C.tropicalis</I> (23%), <I>C.guilliermondi</I> (6%), <I>C.krusei</I> (5%) and others (4%). In the healthy cohort without clinical candidiasis, C.tropicalis was the most commonly isolated species.<br>Conclusion: There appears to be a marked variation in oral <I>Candida</I> species found in HIV-seropositive and seronegative individuals in India. To our knowledge, this is the first attempt to identify oral Candida species in a South Indian population.

Oral lesions of 500 habitual psychoactive substance users in Chennai, India.

The aim of this study was to ascertain the prevalence of oral lesions among 500 psychoactive substance users in a hospital-based population. The study group consisted of 500 consecutive patients attending TTK Hospital, a non-governmental organisation involved in rehabilitation of substance users. Patient history was recorded in a pre-determined format and clinical findings were recorded by a trained physician and dental surgeons. Psychoactive substances used by the patients were alcohol (97%), tobacco (72%), arecanut (57.2%), narcotics (6.8%), cannabis (3.2%) and benzodiazipines (1.8%). Ninety-one percent of patients had one or more oral lesions: dental caries (39%), gingivitis (37.6%), extrinsic stains (24%), oral submucous fibrosis (OSF) (8%), periodontitis (7.4%), leukoplakia (6.6%), melanosis (5.2%), nicotina palatini (2.2%) and erythroplakia (0.6%). For OSF, those using arecanut and alcohol had an odds ratio (OR) of 2.4 [95% confidence intervals (CI) 1.23-4.69, P=0.009], smokers using arecanut products and alcohol had an OR of 3.07 (95% CI 1.59-5.91, P=0.000), and smokers who chewed arecanut products and used drugs had an OR of 23.1 (95% CI 2.05-260, P=0.001) compared with the general population. Those who smoked and used alcohol, arecanut and drugs had a 20.67-fold higher risk of developing leukoplakia compared with those who did not engage in these habits. In conclusion, 91% of patients had one or more oral lesions that needed dental treatment, and most patients were not aware of their oral lesions. The high prevalence of OSF and leukoplakia in substance abusers compared with the general population emphasises the need for regular dental assessments in these patients.