[Optimizing biopsies of the oral mucosa]. / Optimisation des biopsies des muqueuses buccales.

We had for aim to describe and illustrate the artefacts observed in biopsies of the oral mucosa, as well as the impact of sending non-representative histological material to a laboratory. This article was based on an international literature review, as well as on our experience. We analysed the problems raised, for the pathologists and the histology lab-technicians, by these artefacts as well as their impact on the pathology report patient management. We suggest simple solutions.

Bcl-2 expression and its possible influence on malignant transformation of oral lichen planus.

PURPOSE: The aim of this study was to investigate the malignant potential of oral lichen planus (OLP) on the basis of the expression of the Bcl-2 marker in healthy individuals (H), patients with OLP and patients with squamous cell carcinoma (SCC), and to establish potential interdependence between expression of Bcl-2 and the different clinical and histopathological parameters in H, OLP, and SCC. METHODS: Immunohistochemistry for Bcl-2 was carried out using an avidin-biotin peroxidase complex method. Tissue sections were treated with mouse monoclonal antibody against Bcl-2 (124, DAKO A/S, Denmark; dilution 1/100). Immunohistochemical parameters measured included total tissue area, total stained area and intensity of stain. RESULTS: Keratinocytes were not Bcl-2 positive in H and were sparsely positive in OLP. The number of lymphocytes stained with Bcl-2 was significantly lower in H as compared to patients with SCC and OLP. Bcl-2 staining was weak to moderate in OLP, and moderate to intense in SCC. Bcl-2-positive lymphocytes were more expressed in older OLP patients (>55 years), and in OLP specimens with orthokeratinized epithelium, less developed acanthosis and highest grade of lymphocyte expression. CONCLUSION: Bcl-2 may not serve as a prognostic biomarker in oral cancer development from OLP, but it could help in selecting patients with higher need of follow up to prevent malignancy.

Expression of Fas/FasL in patients with oral lichen planus.

PURPOSE: To investigate the malignant potential of oral lichen planus (OLP) on the basis of expression of the Fas/FasL markers in healthy individuals (H), OLP patients and patients with squamous cell carcinoma (SCC). PATIENTS AND METHODS: 40 patients with OLP and two control groups were included in this research (H and patients with SCC). Immunohistochemistry for Fas and FasL was carried out using an avidin-biotin peroxidase complex method. RESULTS: Only a low percentage of infiltrating lymphocytes and no keratinocytes were Fas-positive in OLP specimens. The highest percentage of Fas-staining keratinocytes in our survey was identified mostly in H and patients with well-differentiated SCC. In most cases of SCC, OLP and H a high percentage of keratinocytes and lymphocytes were FasL-positive. FasL expression was negatively correlated with the degree of cell differentiation and apoptosis. Taking into consideration that all carcinomas in this survey were highly differentiated, it is not surprising that no statistically significant differences in FasL expression between H, OLP and SCC specimens were detected. CONCLUSION: Downregulation of Fas expression in keratinocytes and lymphocytes of OLP specimens, together with upregulation of FasL, may serve as initial prognostic biomarker in oral cancer development.

[Genic alterations in oral and head and neck squamous cell carcinomas: analysis of international literature]. / Anomalies géniques dans les carcinomes épidermoïdes des voies aérodigestives supérieures : analyse de la littérature internationale.

The development of oral and head and neck squamous cell carcinomas occurs in relation with multiple events including mainly: loss of cycle cell control, evasion from apoptosis, telomerase reactivation. Complex interactions between a set of molecules, cell cycle proteins, tumour suppressor genes, oncogenes and the telomerase, occur in the multiple step process of carcinogenesis. The 2 main ways of control of the cell cycle rely on 2 tumour suppressor genes: the P53 gene and the retinoblastoma gene or RB gene. One of the regulation pathways or the 2 regulation pathways are disabled during the development of oral and head and neck squamous cell carcinomas. Most of the time, the inactivation of the P53 pathway results from a loss of function of the p53 protein, secondary to mutation and/or deletion of the P53 gene; It may also result of the amplification of the MDM2 gene and of the inactivation of the arf protein. The RB pathway leads to cell proliferation by loss of the p16 protein, by amplification of the cyclin D1 gene and less frequently by mutation of the RB gene or loss of the retinoblastoma protein. In India and South-East Asia, the activation of RAS and MYC oncogenes appears to be related with the presence of specific carcinogens in snuff and tobacco. By blocking apoptosis, the Bcl2 protein seems to increase the resistance of tumours to radiotherapy and chemotherapy.

Nuclear DNA content, an adjunct to p53 and Ki-67 as a marker of resistance to radiation therapy in oral cavity and pharyngeal squamous cell carcinoma.

Factors of prognosis and radioresistance in oral cavity and pharyngeal squamous cell carcinoma (OCPSCC) are limited. In the present study, the usefulness of tumor DNA content in predicting radioresistance in patients with OCPSCC has been investigated. Radioresistance has been defined as local recurrence or tumor persistence after radiation therapy. DNA-ploidy analysis was performed by static cytometry on smears of cell suspensions obtained from formalin-fixed paraffin-embedded material and stained with Feulgen. DNA-ploidy was correlated with the proliferation rate (Ki-67) and p53 protein accumulation obtained by immunohistochemistry. The follow-up of patients ranged from 8 to 62 months. Radioresistance was more common in non-diploid tumors; 14/28 (50%) non-diploid tumors recurred, whereas only 3 (10.7%) out of 28 diploid tumors had local failure (P=0.0019). Proliferation rate and p53 accumulation, evaluated by immunohistochemistry, also added prognostic information. Twelve out of 14 failures were from non-diploid tumors with a low proliferation rate (Ki-67<20%), whereas none of 20 p53-negative diploid tumors developed recurrences. This study showed that non-diploid tumors responded poorly to radiotherapy. DNA content appeared, therefore, as a significant prognostic marker for the evaluation of OCPSCC in patients receiving radiation therapy. This study also showed that DNA content adds information to p53 accumulation and the proliferation rate (Ki-67) for the purposes of determining patient management.

Update on mucous membrane pemphigoid: a heterogeneous immune-mediated subepithelial blistering entity.

Most oral involvement in the skin diseases (dermatoses) is related to mucous membrane pemphigoid or lichen planus. Mucous membrane pemphigoid was the subject of a European Symposium held in Turin, Italy, in June 1997. This review is based on that symposium. Mucous membrane pemphigoid is a subepithelial vesiculobullous disorder mainly of late middle age; it has a slight predilection for women. Whereas mucous membrane pemphigoid was formerly considered a single entity, it is now quite evident that a number of subepithelial vesiculobullous disorders may produce similar clinical pictures and also that a range of variants of mucous membrane pemphigoid exists, with antibodies directed against various hemidesmosomal components or components of the epithelial basement membrane. The term immune-mediated subepithelial blistering diseases has therefore been used. Diagnosis and management of immune-mediated subepithelial blistering diseases on clinical grounds alone are impossible; a full history, general and oral examinations, and biopsy with immunostaining are now invariably required, sometimes supplemented with other investigations. Most patients with mucous membrane pemphigoid affecting the mouth manifest desquamative gingivitis, a fairly common complaint typically seen in women who are middle-aged or older. Oral vesicles and erosions may also occur, and there can be a positive Nikolsky sign. Some patients have lesions of other stratified squamous epithelia, presenting as conjunctival, nasal, oesophageal, laryngeal, vulval, penile, or anal involvement. Apart from improving oral hygiene, immunomodulatory-in particular, immunosuppressive-therapy is typically required to control oral lesions in mucous membrane pemphigoid. No single treatment regimen reliably controls all these disorders.

Oral inflammatory pseudotumour: immunohistochemical investigation of a case involving the submandibular gland and review of the literature.

Inflammatory pseudotumours (IPs) are rare lesions. Most commonly reported in the lung, they are almost ubiquitous, but few oral cases have been described. Their rapid growth, local invasiveness and recurrence, and their ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI) aspects are confusing and mimic benign or malignant neoplasms. Their recognition and distinction from malignant tumors is of importance but their histopathological diagnosis may represent a challenge. In the case reported involving the submandibular gland, the spindle cells had the immunohistochemical profile of myofibroblasts, broader cells with a larger nucleus were CD68 and/or Mac387 positive and the dense plasmacytic infiltrate was polyclonal. Histopathology of IPs covers a spectrum of appearances according to the cellularity and the degree of fibrosis. The recognition of a variable mixture of three main cell types: histiocytes or macrophages, myofibroblasts or fibroblasts and abundant plasma cells, with low mitotic activity and absence of cytological abnormalities in an ill circumscribed and rather fibrous lesion is recommended for the diagnosis of oral IP.

In vitro antifungal resistance of oral Candida albicans strains in non-AIDS patients.

Some cases of oral candidosis are refractory to antifungal treatment. This might be related to development of resistant Candida strains, but susceptibility testing is not standardized and not routinely available, and information related to this problem is scarce in non-AIDS patients. In this study, the in vitro antifungal resistance of oral Candida albicans strains was evaluated. The strains were obtained from a cohort of 72 HIV-negative patients with oral yeast carriage and clinical complaint. Laboratory identification revealed C. albicans in 93% of cases. None of these oral C. albicans isolates showed in vitro resistance to polyenes, but they showed varying resistance levels to fluorocytosine and azoles. This study confirms the usefulness of standardizing susceptibility testing so that it could be routinely available and of realizing a mycological diagnosis including an antifungigram when oral candidosis is suspected, whenever antifungal treatment with azoles is planned.

Markers of radioresistance in squamous cell carcinomas of the head and neck: a clinicopathologic and immunohistochemical study.

PURPOSE: The lack of accurate criteria to predict the response to radiotherapy for individual patients with squamous cell carcinoma of the head and neck (HN-SCC) remains a major problem. The purpose of this study was to investigate the role of several biologic tumor markers to complement clinical prognostic factors in the assessment of response to radiotherapy in SCCs. PATIENTS AND METHODS: p53, ki-67, c-erb B-2, heat-shock protein-27 (HSP-27), and glutathione S transferase (GSTpi) were evaluated by immunohistochemistry on biopsies from 101 patients treated for head and neck cancer by radical radiotherapy. Expression of each marker was correlated with local control and survival using Kaplan-Meier curves. A Cox regression multivariate analysis was also performed that included all clinical and immunohistochemical variables. RESULTS: Expression of p53 and low cell proliferation allowed identification of patients whose tumors did not respond to radiation. Patients with p53-expressing tumors displayed a relative risk (RR) of 3.78 for not being controlled by radiotherapy compared with patients with p53-negative tumors. For tumors with a high growth fraction (ki-67 > 20%) the RR was 0.25 compared with tumors with a low growth fraction (ki-67 < 20%). When p53 expression and cell proliferation were considered simultaneously in a Cox model, the association with resistance to radiation was significant (P = .000004). The RR for resistance with one (p53 staining or ki-67 < 20%) or two (p53 staining and ki-67 < 20%) unfavorable markers was, respectively, 3.8 and 14.87. CONCLUSION: Patients whose tumor expressed p53 with low growth fraction (ki-67 < 20%) had a strong probability not to respond to radiation therapy. Similarly, absence of p53 expression with a high cell proliferation predicted an excellent outcome after radiotherapy even for patients with advanced disease. Prediction of the outcome of radiotherapy would eventually facilitate the early choice of an adequate treatment.

[Buccal dryness, xerostomia and measurement of salivary secretory activity]. / Sécheresse buccale, xérostomie et mesure de l'activité sécrétoire salivaire.

Xerostomia is a marked reduction in saliva production and may occur as an early symptom of various systemic diseases. It is also secondary to the administration of numerous drugs. Severity of salivary gland dysfunction cannot be predicted from subjective reports of oral dryness by patient, and accurate assessments of salivary gland function should be managed. Several procedures for saliva samplings and secretory activity measurements have been reported. In normal individuals, the mean values of unstimulated whole saliva was slightly higher in males (0.42 ml/min) than in females (0.37 ml/min). Corresponding mean values for stimulated whole saliva are 1.77 ml/min and 1.38 ml/min. Unstimulated whole saliva collection during 15 min is recommended as the most reliable test for clinical diagnosis. It is found that an unstimulated salivary flow of 0.1 ml/min represents the cut-off limit for the diagnosis of xerostomia.

Oral carriage of Helicobacter pylori: a review.

Helicobacter pylori is a microaerophilic, motile bacterium, especially adapted to life in the human stomach. The presence of H. pylori in the stomach is strongly associated with chronic gastritis and ulcer disease and is a risk factor for gastric cancers. The microorganism may be transmitted orally and has been detected in dental plaque, saliva, and feces, but the hypothesis that oral microflora may be a permanent reservoir of H. pylori is still controversial. A review of the literature suggests that the recovery of H. pylori in the mouth is probably intermittent, associated with gastroesophageal reflux but not with specific oral disease. Nonetheless, the PCR identification of oral H. pylori may become helpful, particularly in cases of gastritis or ulcer relapse after antimicrobial therapy. Eradication of oral H. pylori by local medication or periodontal procedures would rely on the precise identification of its ecological niche. Within family groups, prophylactic methods should be practiced to avoid oral carriage of H. pylori. The risk of iatrogenic transmission during dental care, however, is already circumscribed by standard professional hygiene procedures.

Oral cryptococcosis: case report of salivary gland involvement in an AIDS patient.

Salivary cryptococcosis was disclosed at autopsy in an AIDS patient with disseminated C. neoformans infection. H & E staining was not suitable to demonstrate the occurrence of C. neoformans in many tissues; Alcian blue gave the best results.

[Drug-induced xerostomia]. / Les hyposialies d'origine médicamenteuse.

Hyposalivation is related to decreased salivary flow, with xerostomia as an ultimate degree. Prolonged severe hyposalivation or xerostomia may induce oral pain, poor tolerance to dentures, loss in taste acuity and increased incidence of oral infections: gingivitis, periodontitis, oral candidosis, infectious sialadenitis and multiple dental caries. Most of the time hyposalivation is a reversible drug-induced side-effect. Hyposalivation is frequent, particularly in elderly people with numerous drugs prescribed on a long-term continuous basis, and in psychiatric patients. It remains a neglected clinical problem. Besides the well-known antimuscarinics, antihistaminics, imipraminic antidepressants and phenothiazic neuroleptics, many drugs may induce hyposalivation. This work aims to review drug-induced xerostomia in 1997 (French pharmacopeae), and high-risk associations.

p53 overexpression in head and neck squamous cell carcinoma: review of the literature.

As a tumour suppressor gene, the inactivation of p53 induces the development of numerous human cancers. Mutations of p53 have been implicated in the pathogenesis of head and neck squamous cell carcinoma (HN-SCC) at a high incidence. In premalignant lesions and in situ carcinomas, p53 overexpression is not exclusively restricted to neoplastic cells, but frequently affects the normal appearing keratinocytes adjacent to p53 positive neoplasms or present in dysplastic areas. These results suggest that as contributors to the early phases of HN-SCC development, p53 alterations may be excellent biomarkers that indicate the predisposition of a particular oral cavity premalignant lesion toward malignancy. In most cases, the p53 overexpression status of a tumour metastasis is identical to that of a primary tumour, indicating that a p53 mutation precedes metastatic spread. In patients with multiple primary tumours, multiple foci of p53 overexpression are observed in epithelia distant from the tumour. So the expression of p53 in normal epithelium would indicate an increased risk for transformation to second or third primary cancers. Distinct p53 mutations in different primary tumours of the same patient indicate that these cancers arise as independent events; these results support the existence of multifocal polyclonal processes. Regardless of the aforementioned results that support p53 as a valid tumour biomarker, most studies have shown no relationship between the expression of p53 and clinical and histopathological parameters. The role played by p53 mutations in the progression and vital prognosis of HN-SCC has not yet been demonstrated.

Histological and ultrastructural study of one case of oral bacillary angiomatosis in HIV disease and review of the literature.

Bacillary angiomatosis (BA) is a new clinicopathological entity defined as a pseudo-neoplastic capillary proliferation secondary to an opportunistic infection by one of two Rochalimaea sp.: R. quintana or R. henselae. Although BA is a recently recognised entity, numerous cases have been reported. Most of the patients affected are reported to have low absolute CD4 lymphocyte counts associated with AIDS. Yet, very few oral cases associated or not with cutaneous lesions have been reported or simply identified. Histopathological and ultrastructural features of one case of oral BA with gingival and palatal lesions are presented. Clinical aspects of oral BA do not hold pathognomonic features and the lesions may resemble either a reactive lesion of the gingiva, pyogenic granuloma or Kaposi's sarcoma. The lesion is characteristically composed of circumscribed lobular capillary proliferations and the presence of granular amphophilic material on haematoxylin and eosin sections surrounded by neutrophils and neutrophilic debris is a clue to diagnosis. Demonstration of bacilli in the interstitium by the Warthin-Starry silver method or, better, by electron microscopy is diagnostic. BA may contribute to the death of the patient but erythromycin has proved to be very effective treatment.

Oral leishmaniasis and Kaposi's sarcoma in an AIDS patient.

In the Mediterranean basin area, visceral leishmaniasis is an endemic disease caused by Leishmania donovani infantum. This study describes the clinical and pathological features of one patient with AIDS who had oral (tonsillar) leishmaniasis, caused by a viscerotropic zymodeme, concurrent with a Kaposi's sarcoma and with a CMV infection.

Epstein-Barr virus DNA detection in gingival tissues of patients undergoing surgical extractions.

The main oral manifestation of Epstein-Barr virus (EBV) infection is hairy leukoplakia, a lesion associated with the acquired immunodeficiency syndrome (AIDS) and occasionally in other immunocompromised patients. However, the recent literature describes the presence of viral genome in clinically normal oral tissues. The purpose of this work was to investigate these occult EBV infections in gingival epithelium. The Southern blot method with 32P-radiolabelled DNA probes under stringent conditions was applied to 20 interproximal gingival papillae specimens and revealed homologous EBV sequences in 4 of 10 AIDS patients as well as in 4 of 10 HIV negative patients. In order to determine whether EBV has a predilection for the gingival tissues, samples of nasal, laryngeal and oral mucosa, other than gingival mucosa, were collected from 10 HIV-negative patients undergoing surgical treatment for a variety of clinical conditions. None of these extra-periodontal mucosal specimens contained homologous EBV DNAs, except an edentulous palatal gingival specimen. With the present detection of EBV DNAs in the gingival tissues of patients undergoing surgical extractions, it would be of interest to investigate more systematically these subclinical infections in order to determine their exact implications in oral disease.

Southern blot detection of human papillomaviruses (HPVs) DNA sequences in gingival tissues.

The highly sensitive and specific methods of molecular biology emphasize the frequency of subclinical infections in the genital tract tissues by the human papillomaviruses (HPVs). The purpose of this work was to investigate occult viral infections by the HPV type 6, 11, 16, and 18 in the gingival tissues. The Southern blot method with 32P-radiolabeled DNA probes applied under stringent conditions to 20 interproximal gingival papilla specimens revealed homologous viral sequences in 1 of 6 cases of adult periodontitis (HPV 16), 1 of 2 cases of rapidly progressive periodontitis (RPP) (HPV 6/HPV 11), 2 of 2 cases of acute gingivitis in psychiatric institutionalized patients (HPV 6; HPV 6/HPV 11), and 2 of 10 cases of acute gingivitis in AIDS patients (HPV 6/HPV 11/HPV 16; HPV 6). No periodontal or extra-periodontal specimen hybridized with the HPV 18 probe. Simultaneous hybridization with two or three HPV types was common (3/6 cases). The present detection of HPV 6, 11, 16 DNAs or related-DNAs in periodontal tissues without obvious clinical signs of viral infection suggests that the gingival epithelium may act as a reservoir.

[Gingival hyperplasia secondary to the use of calcium antagonists: analysis]. / Les hyperplasies gingivales secondaires à la prise d'antagonistes calciques: mise au point.

Calcium antagonists are drugs restricting transmembrane calcium delivery. They possess a wide range of action against vasoconstriction and spastic reactions and were therefore initially recommended for the treatment of angina pectoris. With the increasing number and classes of calcium antagonists new therapeutic indications have emerged. Cases of gingival hyperplasia associated with their use are repeatedly reported, therefore the question deserves to be restated. The aim of the present study was to discuss the clinical, pathologic and pathogenetic features bases on an investigation carried out in a Department of Cardiology, on a case observation and on review of published cases in the international literature.