Marking the Micro Details in Potentially Malignant Disorders of Oral Cavity

Background- The malignancy of the oral cavity constitutes the most important group of malignancies in South and Southeast Asia.The risk increases with the use of tobacco products. Materials & Methods- Micronuclei frequency scoring was used as a biomarker to identify different potentially malignant disorders. Results- Mean micronuclei index was found higher using Hematoxylin and Eosin stain than Papanicolaou's stainand May Grunwald's stain. Conclusion- We concluded that the micronuclei frequencies in oral exfoliated epithelial cells using three different stains- Hematoxylin and Eosin stain, Papanicolaou's stain, May Grunwald's stain may be useful in predicting the malignant potential of premalignant lesions.

Cytomorphologic Analysis of Wet and Spray Fixation Methods in Oral Exfoliative Cytology

@#For a long time, oral exfoliative cytology (OEC) has been implemented as an effective preliminary diagnostic tool for pathological lesions and various methods for fixation of the cytology specimens have been studied. The present study was undertaken to compare the efficacy between the wet and spray type of fixation methods for Papanicolaou (PAP) stained oral cytosmears. The study comprised of 45 healthy subjects in the age group of 20-25 yrs. For each subject, two smears were collected from the buccal mucosa and subjected to wet and spray fixation methods respectively. Both the smears were stained using a commercial Rapid Pap Kit. Smears were observed microscopically and evaluated for cytomorphological features involving uniformity of staining, cellular morphology, nuclear morphology, cellular staining, nuclear staining and presence of impurities. Comparisons were made between the two methods of fixation and statistically analysed using McNemar non-parametric test. Cells were evenly distributed in wet-fixed smears (n=38, 95%) compared to spray fixed smears (n=19, 47.5%). Wet-fixed smears showed lesser impurities (n=13, 32.5%) than spray fixed smears (n=27, 67.5%). However, other parameters such as cytological and nuclear morphology, staining of cytoplasm and nucleus were found to be not significant when compared between the two methods of fixation (p<0.05). The study shows that wet-fixed smears have better cellular distribution and relatively fewer impurities when compared to the spray fixed smears. The method of wet-fixed smears may be used as an alternative to spray fixed smears. A larger sample size may be required for further validation.

A Comparative Study of Micronuclei in Oral Exfoliated Epithelial Cells in Potentially Malignant and Oral Squamous Cell Carcinoma

Background Micronuclei are small, additional nuclei formed as a result of exclusion of chromosome fragments or the whole-chromosome lagging at mitosis. Micronuclei indirectly reflect the chromosomal breakage or impairment of mitotic apparatus. Micronuclei in exfoliated oral epithelial cells are widely used as biomarkers of chromosomal damage, genome instability and cancer risk in humans. Micronuclei scoring can be used as a biomarker to identify different preneoplastic conditions much earlier than manifestations of clinical features and might specifically be exploited in screening of high-risk population for a specific cancer. Aim To correlate frequency of micronuclei in oral exfoliated cells in clinically diagnosed cases of potentially malignant disorders (PMDs) and oral squamous cell carcinoma. Material and Methods The study subjects consisted of clinically and histopathologically diagnosed cases of oral squamous cell carcinoma, oral sub mucous fibrosis and leucoplakia. Healthy subjects without any tobacco consumption habits formed the control group. The cytosmear from all the four groups were stained with Papanicolaou stain. Micronuclei were identified according to the criteria given by Tolbert et al. (1992). Result The frequency of micronuclei was higher in patients with oral squamous cell carcinoma as compared to the other subject groups and the difference was found to be highly significant. Conclusion This study concluded that there is gradual increase in micronuclei counts from normal oral mucosa to PMDs to oral carcinoma.

To evaluate the role of sputum in the diagnosis of lung cancer in south Indian population.

Background: The main advantage of sputum cytology is its simplicity, non-invasiveness and minimal discomfort to the patient. Though, the sputum is evaluated in the diagnosis of lung cancer, the report on the same in the South Indian population was lacking. Therefore, the present study has been undertaken to evaluate the role of sputum in the diagnosis of lung cancer in South Indian population. Methods: The material consisted of sputum samples from 133 patients and was collected in clean wide mouthed disposable plastic containers. Patients were asked to collect sputum the next morning after washing the mouth properly. The sputum was immediately brought to the laboratory and poured into a watch glass. Four smears were prepared from each sample, out of which two smears were immediately fixed in methanol and the other two were air-dried. The methanol fixed smears were stained with Papanicolaou stain. Out of the two air dried smears, one was stained with May Grunwald Giemsa and the other with Gabbot's method for AFB. The smears were screened for malignant cells and a cytological diagnosis was made. The cytological diagnosis was correlated with the histopathological diagnosis. The data obtained were represented as mean percentages. Results: The observation of sputum smears showed numerous pleiomorphic keratinized squamous cells, keratinized squamous cell with hyper chromatic nucleus in well differentiated squamous cell carcinoma, pleiomorphic cells having vacuolated cytoplasm and vesicular nucleus with prominent nucleoli as in adenocarcinoma of the lung, cells arranged in small clusters and having scanty cytoplasm in small cell carcinoma and cells are slightly larger than lymphocyte with scanty cytoplasm and hyper chromatic, grooved nuclei in small cell carcinoma. Conclusion: Cytology of sputum is extremely useful and highly sensitive. The diagnostic accuracy is directly proportional to the number of samples. Sputum cytology is highly sensitive for the centrally located squamous cell carcinoma rather than the peripherally located adenocarcinoma. Properly collected, simple sputum examination alone can give results similar to other highly expensive methods like bronchoscopic material for the diagnosis of lung cancer.

Detection of Trichomonas vaginalis, Gardnerella vaginalis, and Candida Species in Affirm VPIII, Papanicolaou Smear Test and Gram Stain / 대한임상미생물학회지

BACKGROUND: Infectious vaginitis is caused primarily by three different groups of microbial pathogens (Trichomonas vaginalis, Candida spp., and Gardnerella vaginalis). The objective of this study was to compare the Affirm VPIII assay using a DNA hybridization technique with the Papanicolaou (Pap) smear test and the Gram stain in the detection and identification of these three organisms. METHODS: A total of 300 vaginal samples were collected from women that were either symptomatic for vaginitis or asymptomatic women that were being seen for routine obstetric or gynecological care. The presence of T. vaginalis, Candida spp., and G. vaginalis was evaluated by using the Affirm VIII assay (Becton Dickinson, USA), Pap smear test, and Gram stain method, respectively. RESULTS: With the Affirm VPIII assay, 1 (0.3%) patient tested positive for T. vaginalis, 99 (33.0%) patients were positive for G. vaginalis, and 18 (6.0%) were positive for Candida spp. The detection rates of Trichomonas infection, bacterial vaginosis and candidiasis by the Pap smear test and Gram stain method were 0.7% versus 0%, 16.3% versus 35.7%, and 1.7% versus 9.7%, respectively. The differences between the detection rates of the above three organisms between the Pap smear test and the Gram stain method were statistically significant (p<0.05). CONCLUSION: The Affirm VPIII assay was more sensitive than the Pap smear test and more specific than the Gram stain method for the detection and identification of these three organisms. In addition, the results of the Affirm VPIII assay are quick to obtain and are simple and easy to interpret.

Detection for Chlamydia trachomatis by Vidas Chlamydia Test and Direct Smear / 감염

BACKGROUND: Because Chlamydia trachomatis infection is the most frequent etiological agent in sexually transmitted diseases, accurate diagnostic methods are essential. The enzyme-linked fluorescence assay and Papanicolaou smear were evaluated for detection of C. trachomatis in patients with vaginitis. METHODS: Endocervical swabs from 78 women with symptoms of vaginal discharge, itching or lower abdominal pain in the obstetrics/gynecology department of Gyeongsang National University Hospital were tested by Vidas Chlamydia assay. Slides were stained with Papanicolaou and Giemsa stain to demonstrate inclusion body. RESULTS: Five (6.4%) of 78 endocervical specimens were positive by Vidas Chlamydia assay. Only one case was positve for C. trachomatis with Papanicolaou stain. We could not detect any C. trachomatis by Giemsa stain. CONCLUSION: The Vidas Chlamydia test allows sensitive and high-volume testing for chlamydiae and could be useful for the diagnosis of infection with C. trachomatis in patients with vaginitis and pelvic inflammatory disease.

Sperm Morphology Analysis Using Strict Criteria in Assessing Normal and Amorphus Sperm / 대한남성과학회지

PURPOSE: We evaluated the validity and reproducibility of sperm morphology analysis using strict criteria to compare the difference in the percentages of normal and amorphous morphology as seen by two staining methods (Diff-Quik and Papanicolaou)with the eventual goal of developing a computer-assisted sperm morphology assessment system. MATERIALS AND METHODS: Semen samples were obtained from 10 volunteers and stained by the Diff-Quik and Papanicolaou staining methods. Under 1000-fold oil-immersion magnification, the slides were examined by two experienced observers, and the screens were transferred to the monitor using a CCD camera. A calibrated transparent overlay was used to distinguish between normal and abnormal morphology according to strict criteria. One hundred spermatozoa for each sample were analyzed by two experienced observers. RESULTS: In Diff-Quik staining, intraobserver and interobserver correlation coefficients for normal and amorphous sperm were 0.862 (p<0.05), 0.916 (p<0.05) and 0.857 (p<0.05), 0.890 (p<0.05), respectively. In Papanicolaou staining, intraobserver correlation coefficients for normal and amorphous sperm were 0.832 (p<.05), 0.877 (p<0.05) and 0.846 (p<0.05) and 0.865 (p<0.05), respectively. CONCLUSIONS: Assessing normal and abnormal spem abnormal sperm morphology using strict criteria produces reproducible results between and within observers. Also, there were good correlations between the two staining methods in assessing normal and amorphous morphology. This study presents the basic data for the development of computer-assisted sperm morphology assessment system. New techniques should be developed to analyze amorphous sperm.

Sperm Morphology Analysis Using Strict Criteria in Assessing Normal and Amorphus Sperm / 대한남성과학회지

PURPOSE: We evaluated the validity and reproducibility of sperm morphology analysis using strict criteria to compare the difference in the percentages of normal and amorphous morphology as seen by two staining methods (Diff-Quik and Papanicolaou)with the eventual goal of developing a computer-assisted sperm morphology assessment system. MATERIALS AND METHODS: Semen samples were obtained from 10 volunteers and stained by the Diff-Quik and Papanicolaou staining methods. Under 1000-fold oil-immersion magnification, the slides were examined by two experienced observers, and the screens were transferred to the monitor using a CCD camera. A calibrated transparent overlay was used to distinguish between normal and abnormal morphology according to strict criteria. One hundred spermatozoa for each sample were analyzed by two experienced observers. RESULTS: In Diff-Quik staining, intraobserver and interobserver correlation coefficients for normal and amorphous sperm were 0.862 (p<0.05), 0.916 (p<0.05) and 0.857 (p<0.05), 0.890 (p<0.05), respectively. In Papanicolaou staining, intraobserver correlation coefficients for normal and amorphous sperm were 0.832 (p<.05), 0.877 (p<0.05) and 0.846 (p<0.05) and 0.865 (p<0.05), respectively. CONCLUSIONS: Assessing normal and abnormal spem abnormal sperm morphology using strict criteria produces reproducible results between and within observers. Also, there were good correlations between the two staining methods in assessing normal and amorphous morphology. This study presents the basic data for the development of computer-assisted sperm morphology assessment system. New techniques should be developed to analyze amorphous sperm.