Aetiology and therapeutics of burning mouth syndrome: an update

doi: 10.1111/j.1741‐2358.2010.00384.x Aetiology and therapeutics of burning mouth syndrome: an update Objective: To provide a review on the aetiology and therapeutic options for the management of patients with burning mouth syndrome (BMS). Background: BMS is a chronic disorder that frequently affects women and is characterised by burning symptoms of the oral mucosa without clinical signs. This syndrome has a complex and multifactorial characteristics, but its aetiology remains unknown and this makes it difficult with regard to the treatment and management of such patients. Despite not being accompanied by evident organic changes and not presenting risks to health, BMS can significantly reduce the quality of life for patients. Methods and materials: The article reviews the literature regarding aetiologic factors, clinical implications and treatment of BMS. Conclusion: The involvement of neurological, emotional and hormonal alterations is proposed in BMS aetiology. However the mechanisms of its development are complex and not completely understood. Tricyclic antidepressants, benzodiazepines and antipsychotic drugs are the most accepted options in treatment and show variable results. The correct diagnosis of BMS and the exclusion of possible local or systemic factors that can be associated with the symptoms are fundamental. It is also important to evaluate the quality of life for these patients to recognise the potential impact of this condition on their lives.     

Burning mouth syndrome (BMS): sialometric and sialochemical analysis and salivary protein profile

Objective: The aim of the present study was to analyse the characteristics of salivary production and its composition in individuals with burning mouth syndrome (BMS). Study Design: Salivary flow rate, concentrations of potassium, iron, chloride, thiocyanate, magnesium, calcium, phosphorus, glucose, total protein and urea, as well as the expression profile of salivary proteins were analysed by SDS‐PAGE. Results: The mean salivary flow rate among control patients was lower than that of BMS patients. Chloride, phosphorus and potassium levels were elevated in patients with BMS (p = 0.041, 0.001 and 0.034, respectively). Total salivary protein concentration was reduced in individuals with BMS (p = 0.223). Analysis of the expression of salivary proteins by Coomassie blue SDS‐PAGE revealed a lower expression of low molecular weight proteins in individuals with BMS compared to healthy controls. Conclusions: These results indicate that the identification and characterisation of low molecular weight salivary proteins in BMS may be important in understanding BMS pathogenesis, thus contributing to its diagnosis and treatment.     

Usefulness of new wetness tester for diagnosis of dry mouth in disabled patients

Objectives: The condition of dry mouth is an influential factor in the incidence of caries, periodontal disease, fungal infections, masticatory dysfunctions and denture function. Bedridden elderly and disabled persons often suffer from oral dryness and the aim of this study was to evaluate the usefulness of measuring the amount of moisture in the oral mucosa for clinical diagnosis of dry mouth in this group. Subjects and methods: The subjects were 20 elderly bedridden individuals, age range 65–89 years old, living in a nursing home and six healthy laboratory researchers, aged 20–46 years old, used as controls. Tongue dorsum moisture measurements were performed using a newly developed wetness tester (L‐SALIVO^®), in which the wet portion was measured after 10 s. Further, clinical diagnosis of dry mouth was carried out using a clinical classification scale of the tongue mucosa (grade range, 0–3). Results: It was possible to measure tongue dorsum moisture in all subjects with the wetness tester. The average moisture value was 0.1 ± 0.2 mm in elderly subjects with a dry mouth grade of 2 (n = 8) or 3 (n = 12), while the average moisture value in the control subjects was 3.67 ± 1.75 mm with a dry mouth grade of 0 (n = 4) or 1 (n = 2). Tester values and clinical classification showed a positive co‐relationship (r = 0.31, p < 0.05). Conclusions: Our results show that this new tester could be useful for evaluating oral dryness and diagnosing dry mouth.     

A morphometric study of the protective effect of cryotherapy on oral mucositis in cancer patients treated with 5-fluorouracil

We investigated cytological changes in oral mucosa smears from patients treated with cryotherapy to determine whether cryotherapy prevented mucositis caused by 5-fluorouracil (5-FU) therapy. Patients with gastrointestinal malignancies were divided into four groups; control patients before 5-FU therapy, patients after 5-FU therapy without cryotherapy, patients with cryotherapy before 5-FU therapy and patients with cryotherapy after 5-FU therapy. Oral mucosa samples from all patients were assessed at the beginning and on day 14 of chemotherapy. We used exfoliative cytology to evaluate cellular changes in the oral mucosa that were caused by 5-FU. Smears from each patient were stained using the Papanicolaou method and analyzed using stereology. Smears were taken from each group before and after 5-FU infusion. We found that nuclear volume was decreased significantly in cells of the 5-FU therapy after cryotherapy patients compared to the 5-FU therapy before cryotherapy patients. We also found significantly decreased cytoplasmic volumes in the 5-FU therapy after cryotherapy patients compared to the 5-FU therapy before cryotherapy patients. The results of cytomorphometric estimations revealed that cryotherapy may be used to prevent damage to oral tissue and may decrease the frequency and duration of oral mucositis caused by 5-FU.     

Cytologic analysis of alterations induced by Smoking and by alcohol consumption

OBJECTIVE: To analyze cytologically the buccal mucosa of smoking and nonsmoking volunteers to determine what cellular changes are induced by cigarettes and alcohol consumption. STUDY DESIGN: In order to evaluate cellular changes induced by smoking and alcohol consumption, exfoliative cytology was used for the analysis of mucosal smears obtained from the buccal mucosa of 25 smokers and 25 nonsmokers. The number of cigarettes consumed, duration of smoking, presence or absence of alcohol ingestion, ingested alcohol dose and frequency of consumption, and most frequently used type of alcoholic beverage were determined using a questionnaire. Three smears from each individual stained by the Papanicolaou method were analyzed quantitatively and qualitatively under a light microscope by 2 experienced examiners in terms of inflammatory and dysplastic alterations and of the degree of epithelial maturation. RESULTS: Although numerous alterations were observed in smokers they corresponded up to only Papanicolaou class II and were not significantly different from nonsmokers (Mann-Whitney and chi2 tests, p < 0.05). A higher proportion of inflammatory cells (polymorphonuclear and mononuclear cells) were obtained from smokers as compared to nonsmokers, while the proportion of bacteria was similar in the 2 groups. CONCLUSION: The findings indicate that even after a short period of cigarette use and alcohol consumption, inflammatory alterations were detectable on exfoliative cytology of the buccal mucosa in a young group, demonstrating the usefulness of cytology for early detection in smokers.     

Cytologic and DNA-cytometric early diagnosis of oral cancer.

OBJECTIVE: The aim of this prospective study was to report on the diagnostic accuracy of conventional oral exfoliative cytology taken from white-spotted, ulcerated or other suspicious oral lesions in our clinic. In addition we checked DNA-image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA-aneuploidy is a sensitive and specific marker for the early identification of tumor cells in oral brushings. STUDY DESIGN: 251 cytological diagnoses obtained from exfoliative smears of 181 patients from macroscopically suspicious lesions of the oral mucosa and from clinically seemingly benign oral lesions which were excised for establishing histological diagnoses were compared with histological and/or clinical follow-ups of the respective patients. Additionally nuclear DNA-contents were measured after Feulgen restaining using a TV image analysis system. RESULTS: Sensitivity of our cytological diagnosis on oral smears for the detection of cancer cells was 94.6%, specificity 99.5%, positive predictive value 98.1% and negative predictive value 98.5%. DNA-aneuploidy was assumed if abnormal DNA-stemlines or cells with DNA-content greater 9c were observed. On this basis the prevalence of DNA-aneuploidy in smears of oral squamous cell carcinomas in situ or invasive carcinomas was 96.4%. Sensitivity of DNA-aneuploidy in oral smears for the detection of cancer cells was 96.4%, specificity 100%, positive predictive value 100% and negative 99.0%. The combination of both techniques increased the sensitivity to 98.2%, specificity to 100%, positive predictive value to 100% and negative to 99.5%. CONCLUSIONS: Brush cytology of all visible oral lesions, if they are clinically considered as suspicious for cancer, are an easily practicable, cheap, non-invasive, painless, safe and accurate screening method for detection of oral precancerous lesions, carcinoma in situ or invasive squamous cell carcinoma in all stages. We conclude that DNA-image cytometry is a very sensitive, highly specific and objective adjuvant tool for the early identification of neoplastic epithelial cells in oral smears.     

Cytobrush and wooden spatula for oral exfoliative cytology. A comparison.

The Cytobrush has been used frequently in cervical cytology, but as yet its value in oral exfoliative cytology has not been assessed. A study was undertaken to compare the efficiency of the Cytobrush with that of the wooden tongue spatula. For 26 patients, two smears were collected from clinically normal mucosa from four sites in the oral cavity (buccal mucosa, dorsal tongue, ventral tongue and hard palate). The smears were graded for cell yield and dispersion on a three-point scale. The results were analyzed using the chi 2 test. The Cytobrush was found to be significantly more efficient than the wooden spatula, in terms of both cell yield (P less than .005) and cell dispersion (P less than .005). When each site was examined separately, the Cytobrush produced significantly better dispersion for the dorsal tongue, ventral tongue and buccal mucosa and a better cell yield for the tongue surfaces. No significant difference for cell yield or dispersion was found for the hard palate. The study showed that the Cytobrush is an effective instrument for use in exfoliative cytology of normal oral mucosa.     

Prediction of recurrence using exfoliative cytology and melanoma‐associated antigen‐A mRNA analysis following wide excision of oral squamous cell carcinoma: short report

N. Mollaoglu, P. Metzler, J. Zenk, E. Nkenke, F. W. Neukam and J. Ries
Prediction of recurrence using exfoliative cytology and melanoma‐associated antigen‐A mRNA analysis following wide excision of oral squamous cell carcinoma: short report Background: Oral squamous cell carcinoma (OSCC) is the sixth most common cancer. The local recurrence of OSSC might result from the existence of occult cancer cells around tumour margins. Exfoliative cytology has lately gained great importance as a method for obtaining RNA samples from suspicious oral mucosal lesions in order to carry out molecular diagnosis. In addition, melanoma associated‐A antigens (MAGE‐A) are expressed in various tumours and their detection is a highly accurate sign that cancer cells are present. Objective: The prediction of a recurrence using MAGE‐A mRNA expression analysis to follow‐up OSCC cases using a newly established molecular diagnostic technique applied to cytological materials. Methods: RNA was extracted from three recurrent OSCC cases and from 20 healthy volunteers as a control group using a cytobrush. The expression of MAGE‐A3, A4, A6, A10 and A12 was investigated in these specimens using quantitative real‐time (RT‐PCR). Results: There was no expression of MAGE‐A in the specimens of normal oral mucosa. However, the expression analysis of five different MAGE‐A genes indicated a high potential for malignant change in biopsy‐proven recurrent OSCC cases. Except for MAGE‐A10, the rest of the genes were expressed in different ratios by the three recurrent cases, which had been determined on histopathology to be OSCC or carcinoma in situ. Conclusion: It is suggested that analysis of MAGE‐A expression may be used as a risk prediction method in the diagnosis of recurrence after wide excision of OSCC to enhance the accuracy of exfoliative cytology, which has limitations due to false negative and false positive results.     

Salivary tests associated with elderly people’s oral health

doi: 10.1111/j.1741‐2358.2012.00627.x Salivary tests associated with elderly people’s oral health Introduction: The saliva constitutes essential condition for the individual’s health. Aim: Identify the relation of the salivary flow and saliva pH with medicine use and oral discomfort in elderly. Methods and materials: Cross‐sectional study with 68 elderly living in a long staying institution. Salivary tests were performed based on Bo Krasse’s methodology. For pH, the Universalindikator – Merck tape was used. A questionnaire was applied, organising data through Software SPSS version 17. Pearson’s qui‐square distribution, Fisher’s exact test and t‐test for paired data were used, with significance level of 5% and confidence interval of 95%. Results: Among the 68 elderly (average of 70.4 years, SD ± 7.27), 80% showed normal pH. The rate of salivary flow was as follows: very low, 32.3%; lowered, 41.2%; and normal, 25.5%; 30.9% reported dry mouth; 22.1% problems with taste; 17.6%, dysphagia; and 14.7%, burning mouth. 76.5% used medicines. There was statistical significance between medicine use and dry mouth (p = 0.015). They showed an association between salivary flow and medicine use (p = 0.048), feels dry mouth (0.018) and difficulty to swallow (p = 0.046), and saliva pH without stimulation and feels dry mouth (p = 0.003), difficulty to swallow (p = 0.006) and burning mouth sensation (p = 0.014). Conclusion: Low salivary flow and saliva pH interfere on elderly people’s health and medicine use influences on results.     

Quantitative analysis of AgNOR proteins in exfoliative cytology specimens of oral mucosa from smokers and nonsmokers

OBJECTIVE: To determine the cell proliferation rate and possible effects of cigarette smoking on the oral mucosa lining through analysis of silver-stained nucleolar organizer regions (AgNORs) in exfoliative cytology specimens. STUDY DESIGN: Exfoliative cytology was performed on the left side of the border of the tongue and of the floor of the mouth in 25 smoking patients and 25 nonsmoking patients. The inclusion criterion for smokers was the consumption of more than 20 cigarettes per day for a minimum of 30 years. RESULTS: The slides were stained by histochemical AgNOR method. In the nonsmoking group the mean number of AgNORs per nucleus was 2.732 +/- 0.236 in the tongue border and 2.918 +/- 0.195 in the floor of the mouth. In smoking patients the mean number of AgNORs per nucleus was 3.372 +/- 0.375 in the tongue border and 3.245 +/- 0.237 in the floor of the mouth. CONCLUSION: The results suggest higher cell proliferation quantified by the histochemical AgNOR technique in exfoliative cytology specimens obtained from the oral mucosa lining of smokers presenting no clinical alterations.     

Comparison of the Cytobrush®, dermatological curette and oral CDx® brush test as methods for obtaining samples of RNA for molecular analysis of oral cytology

M. D. Reboiras‐López, M. Pérez‐Sayáns, J. M. Somoza‐Martín, P. Gayoso‐Diz, F. Barros‐Angueira, J. M. Gándara‐Rey and A. García‐García Comparison of the Cytobrush^®, dermatological curette and oral CDx^® brush test as methods for obtaining samples of RNA for molecular analysis of oral cytology Objective: Interest in oral exfoliative cytology has increased with the availability of molecular markers that may lead to the earlier diagnosis of oral squamous cell carcinoma. This research aims to compare the efficacy of three different instruments (Cytobrush, curette and Oral CDx brush) in providing adequate material for molecular analysis. Methods: One hundred and four cytological samples obtained from volunteer healthy subjects were analysed using all three instruments. The clinical and demographical variables under study were age, sex and smoking habits. The three instruments were compared for their ability to obtain adequate samples and for the amount of RNA obtained using quantitative real‐time polymerase chain reaction (PCR‐qRT) analysis of the Abelson (ABL) housekeeping gene. Results: RNA of the ABL gene has been quantified by number of copies. Adequate samples were more likely to be obtained with a curette (90.6%) or Oral CDx (80.0%) than a Cytobrush (48.6%); P < 0.001. Similarly, the RNA quantification was 17.64 ± 21.10 with a curette, 16.04 ± 15.81 with Oral CDx and 6.82 ± 6.71 with a Cytobrush. There were statistically significant differences between the Cytobrush and curette (P = 0.008) and between the Cytobrush and OralCDx (P = 0.034). There was no difference according to the demographical variables. Conclusions: Oral exfoliative cytology is a simple, non‐invasive technique that provides sufficient RNA to perform studies on gene expression. Although material was obtained with all three instruments, adequate samples were more likely to be obtained with the curette or Oral CDx than with a Cytobrush. The Oral CDx is a less aggressive instrument than the curette, so could be a useful tool in a clinical setting.     

Cytologic and cytometric analysis of oral mucosa in Alzheimer's disease

OBJECTIVE: To analyze cytomorphologically the buccal mucosa of patients with Alzheimer's disease (AD). STUDY DESIGN: Brush biopsies were obtained from 10 patients with AD and 9 age-matched controls without neurologic symptoms from 3 distinct oral sites. RESULTS: A significant reduction in partially keratinized intermediate (red) cells was observed in the buccal mucosa of the AD group. In the AD group, parabasal cells from the floor of the mouth (p = 0.017) and buccal mucosa (p = 0.058) and red cells,from the tongue dorsum (p = 0.013) and buccal mucosa (p = 0.002), exhibited significantly greater nuclear areas. With regard to the nuclear to cytoplasmic (N:C) ratio, intermediate (red) cells from the buccal mucosa and tongue dorsum of AD individuals showed a decrease in this parameter (p <0.0001), while superficial (yellow) cells (from buccal mucosa) (p= 0.042) and parabasal (blue) cells (from the tongue dorsum) (p = 0.003) exhibited an increased N:C ratio. No significant differences were detected in the cells from the floor of the mouth. CONCLUSIONS: Our findings indicate that cytologic and cytometric changes were detectable in the exfoliative cytology of the buccal mucosa and tongue in the AD group.     

Exfoliated oral mucosa cells as bioindicators of short- and long-term systemic titanium contamination

BackgroundHumans are exposed to exogenous sources of titanium-containing particles that can enter the body mainly by inhalation, ingestion, or dermal absorption. Given the widespread use of biomaterials in medicine, the surface of a titanium (Ti) biomedical device is a potential endogenous source of Ti ions and/or Ti-containing particles, such as TiO₂ micro-(MPs) and nano-particles (NPs), resulting from biotribocorrosion processes. Ti ions or Ti-containing particles may deposit in epithelial cells of the oral mucosa, and the latter may therefore serve as bioindicators of short and long-term systemic Ti contamination. The aim of the present study was to histologically and quantitatively evaluate the presence of Ti traces in cells exfoliated from the oral mucosa as possible bioindicators of systemic contamination with this metal at short and long-term experimental time pointsMethodsThirty Wistar rats were intraperitoneally injected with a suspension of titanium dioxide (TiO₂) (0.16 g/100 g body weight of TiO₂ in 5 ml of NaCl 0.9%) using 5 nm NPs (Group: TiO₂-NP5; n = 10), 45 µm MPs (Group: TiO₂-MP45; n = 10), or vehicle alone (Control group; n = 10). At one and six months post-injection, right-cheek mucosa cells were obtained by exfoliative cytology using a cytobrush; they were spray fixed and stained using Safranin or the Papanicolaou technique. The smears were cytologically evaluated (light microscopy) to determine the presence of particulate material, which was also analyzed microchemically (SEM-EDS). Left-cheek mucosa cells were similarly obtained and re-suspended in 5 ml of PBS (pH: 7.2–7.4); the samples corresponding to each group were pooled together and analyzed spectrometrically (ICP-MS) to determine Ti concentration in each of the studied groups. Blood samples were obtained for histological determination of the presence of particulate material on Safranin-stained blood smears and determination of plasma concentration of Ti by ICP-MSResultsDifferent size and shape metal-like particles were observed inside and outside epithelial cells in TiO₂-NP5 and TiO₂-MP45 cytological smears at both one and six months post-injection. EDS analysis showed the presence of Ti in the particles. ICP-MS revealed higher Ti concentrations in both TiO₂ injected groups compared to the control group. In addition, Ti concentration did not vary with time or particle size. Monocytes containing particles were observed in blood smears of TiO₂-exposed animals one- and six-months post-injection. Plasma levels of Ti were significantly higher in TiO₂-NP5- and TiO₂-MP45- exposed animals than in controls (p < 0.05), and Ti concentration was significantly higher at one month than at six months in both TiO₂-exposed groups (p < 0.05).ConclusionsCells exfoliated from the oral mucosa could be used as bioindicators of short- and long-term systemic contamination with Ti. Exfoliative cytology could be used as a simple, non-invasive, and inexpensive diagnostic method for monitoring biotribocorrosion of Ti implants and patient clinical follow-up.     

Significance of saliva for the denture‐wearing population

This paper summarises a series of studies already published in German and presents new data related to the aetiology of the dry mouth' and its associated problems. Aims: to study factors affecting mucous and serous salivary gland secretion, the aetiology of the ‘dry mouth’ and its associated problems, causative factors for hyposalivation and it's treatment Setting : two university dental hospitals. Subjects: 587 denture wearers and 521 control subjects, and autopsy material Interventions : exercise, chewing, water, oestrogen, pilocarpine, and anetholtrithion theiapy, biopsy of the minor glands Main outcome measures : Palatal secretion (PAL, μL/cm²/min) and parotid salivary flow (PAR), subjective complaints and clinical findings. Results: resting flow rates for PAL between 0 and 65 μl/cm²/min were seen in every age group. The flow rates of PAR (0 to 3.7 ml/10 min) were not correlated with PAL. Most patients with a resting flow rate of PAL≤6.0 μl/cm² suffer from a ‘dry mouth’ and Burning Mouth Syndrome (BMS) or oral dysaesthesia (OD) with or without chronic lesions of the oral mucosa. Etiological factors for the incidence of reduced PAL and associated problems include xerostomic drugs, oestrogen deficiency, ladiotherapy, thyroid dysfunction, smoking or continuous wearing of complete upper dentures. PAL also correlated with the retention of upper complete dentures. PAL was correlated with the water content of epithelial tissues. PAL and PAR were both increased by drinking ample fluid, improving their circulation by physical exercises, chewing intensively, or taking oestrogens, pilocarpine, anetholtrithion. Conclusions: Variation in palatal salivary secretion occurs and is clinically important.     

Diagnostic value of nucleolar organizer regions (AgNORs) in brush biopsies of suspicious lesions of the oral cavity.

OBJECTIVE: The aim of this retrospective study was to report on the diagnostic accuracy of AgNOR-analysis as an adjunctive diagnostic tool of conventional oral exfoliative cytology taken from suspicious lesions in our clinic. STUDY DESIGN: Cytological diagnoses obtained from brush biopsies of macroscopically suspicious lesions of the oral mucosa from 75 patients (final diagnoses: 53 histologically proven squamous cell carcinomas, 11 leukoplakias and other inflammatory oral lesions) and from 11 patients with normal mucosa as a negative control group were compared with histological and/or clinical follow-ups. Five smears were doubtful and seven suspicious for tumor cells in the cytologic report. Number of AgNOR's were counted in 100 squamous epithelial cell-nuclei per slide after silver-restaining. RESULTS: Sensitivity of our cytological diagnosis alone on oral smears for the detection of squamous carcinomas was 92.5%, specificity 100%, positive predictive value was 100% and negative 84.6%. The best cut-off value of the mean number of AgNOR dots per nucleus distinguishing benign from malignant cells was 4.8. The percentage of nuclei with more than three AgNORs had a cut-off level of 70%. Applying these methods to twelve doubtful or suspicious cytological diagnoses we were able to correctly establish the diagnosis of malignancy in ten cases of histologically proven cancers and to reveal benignity in two histologically proven cases. Thus we achieved a positive and negative predictive value of 100% each. CONCLUSIONS: Smears from brushings of visible oral lesions, if clinically considered as suspicious for cancer, are an easily practicable, non-invasive, painless, safe and accurate screening method for detection of oral cancerous lesions. We conclude that AgNOR-analysis may be a useful adjunct to other methods in routine cytological diagnosis of oral cancer that can help to solve cytologically suspicious or doubtful cases.     

Immunocytochemical detection of Ki‐67 in Diff‐Quik‐stained cytological smears of canine mammary gland tumours

U.S. Choi and D.Y. Kim Immunocytochemical detection of Ki‐67 in Diff‐Quik‐stained cytological smears of canine mammary gland tumours Objective: To investigate whether Diff‐Quik stained fine needle aspirate smears can be used to evaluate Ki‐67 expression by immunocytochemistry. Methods: Both cytological and histological samples were obtained from 24 dogs with spontaneously developed mammary gland tumours. The cytological and histological specimens were examined by Diff‐Quik and H&E stains, respectively. After examination, both samples were immunostained using the same Ki‐67 antibody. The % Ki‐67 values were calculated based on the percentage of positively stained tumour cells per 500 and 1000 tumour cells in cytology and histology specimens, respectively. Results: Ki‐67 staining was successful in 17/24 smears (71%) and 19/23 sections (83%). The correlation coefficient between the percentage of Ki‐67‐positive cells in cytological smears and in the histological sections was 0.677 (P < 0.01). These values were significantly different between histologically benign and malignant tumour groups both in cytology and histology samples (P < 0.001). The threshold value of the percentage of Ki‐67‐positive cells for distinguishing benign from malignant tumours was set at 4.85% with 90.9% sensitivity and 92.3% specificity by Receiver Operating Characteristic (ROC) curve using histopathology as the gold standard. Conclusion: Diff‐Quik‐stained cytology smears can be used to detect the presence of Ki‐67 antigen when histology sections are not available.     

Analysis of silver binding nucleolar organizer regions in exfoliative cytology smears of potentially malignant and malignant oral lesions

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation.     

Quantitative assessment of various fixatives on nuclear and cytoplasmic areas in oral cytology

Summary Both nuclear and cytoplasmic areas are parameters known to be of significance in the diagnosis of malignancy. However, few studies have assessed the effect of fixation on exfoliative cytology and none has looked at such influences upon oral smears. Hence the method of fixation may influence directly diagnostic cytology. The effect of three methods of fixation upon the nuclear and cytoplasmic areas of cells removed from the buccal mucosa was quantitatively assessed. The three methods employed, prior to Papanicolaou staining, were: direct immersion in diethylether and ethanol (11 v/v), spray fixation (Vale Smear Fix) and air drying. Three smears from each of 21 patients were used, each slide being allocated randomly a, method of fixation. After 24h all smears were processed for Papanicolaou's stain.The nuclear and cytoplasmic areas were calculated using semi-automated image analysis. No significant differences were found in the two areas whichever method of fixation was used.     

Natural Killer Cell Receptor+ T‐Lymphocytes in Normal and Helicobacter pylori‐Infected Human Gastric Mucosa

Background: Helicobacter pylori infection is associated with development of chronic inflammation and infiltration of immune cells into the gastric mucosa. As unconventional T‐lymphocytes expressing natural killer cell receptors are considered to play central roles in the immune response against infection, a study investigating their frequencies in normal and H. pylori‐infected gastric mucosa was undertaken. Materials and Methods: Flow cytometry was used to quantify T‐cells expressing the natural killer cell markers CD161, CD56, and CD94 in freshly isolated lymphocytes from the epithelial and lamina propria layers of gastric mucosa. Thirteen H. pylori‐positive and 24 H. pylori‐negative individuals were studied. Results: CD94⁺ T‐cells were the most abundant (up to 40%) natural killer receptor‐positive T‐cell population in epithelial and lamina propria layers of H. pylori‐negative gastric mucosa. CD161⁺ T‐cells accounted for about one‐third of all T‐cells in both compartments, but the lowest proportion were of CD56⁺ T‐cells. Compared with H. pylori‐negative mucosa, in H. pylori‐infected mucosa the numbers of CD161⁺ T‐cells were significantly greater (p = .04) in the epithelium, whereas the numbers of CD56⁺ T‐cells were lower (p = .01) in the lamina propria. A minor population (< 2%) of T‐cells in both mucosal layers of H. pylori‐negative subjects were natural killer T‐cells, and whose proportions were not significantly different (p > .05) to those in H. pylori‐infected individuals. Conclusions: The predominance, heterogeneity, and distribution of natural killer cell receptor‐positive T‐cells at different locations within the gastric mucosa reflects a potential functional role during H. pylori infection and warrants further investigation.     

The impact of liquid-based oral cytology on the diagnosis of oral squamous dysplasia and carcinoma

OBJECTIVE: Even though diagnostic oral exfoliative cytology is a useful, economical and practical tool in the diagnosis of oral dysplasia and carcinoma, it is not yet extensively used. The results of conventional exfoliative and liquid-based diagnostic cytology in oral potentially malignant lesions (PML) are herein reported and compared with the histological diagnosis. METHODS: Either conventional (89) or liquid-based (384) exfoliative cytology was used for the diagnosis of oral dysplasia/carcinoma in 473 subjects and the results were compared with scalpel biopsy histology. Cells were collected using a Cytobrush device for conventional smears and with a dermatological curette for the liquid-based cytology. The 'curette technique' also allowed for the collection of 'accidental' tissue fragments, utilized as microbiopsies. RESULTS: Histological diagnosis was squamous carcinoma in 96 of 473 cases, high-grade dysplasia (oral intraepithelial neoplasia two to three) in 24 and other lesions in 353 cases. The smears in the conventional cytology group were inadequate in 12.4%, with an 85.7% sensitivity and a 95.9% specificity. There were 8.8% of inadequate specimens in the liquid-based cytology group; sensitivity was 95.1% and specificity was 99.0%. CONCLUSIONS: Although conventional cytology is useful when diagnosing oral PML (better sensitivity and predictive positive value if compared with the cervical smear test with similar specificity) and can improve the accuracy of histological diagnosis, liquid-based cytology gives better results, as it not only enhances both sensitivity and specificity, but also provides material for further investigation (AgNORs, DNA, microbiopsies, etc.).