Detection and typing of human papillomaviruses (HPV) in malignant, dysplastic, nondysplastic and normal oral epithelium by nested polymerase chain reaction, immunohistochemistry and transitional electron microscopy in patients of northern Greece.

OBJECTIVES: To evaluate the role of HPV in oral carcinogenesis, we examined the prevalence of HPV in malignant, potentially malignant and normal oral epithelium and studied the relation of HPV prevalence with other factors obtained from the patient's records. MATERIALS AND METHODS: Our material consisted of 291 tissue specimens from 258 individuals. From every individual formalin fixed and paraffin embedded tissues were examined by nested Polymerase Chain Reaction (NPCR) for the detection of HPV DNA and by immunohistochemistry (IHC) for the in situ detection of HPV L1 protein. Positive PCR products were sequenced in order to type HPVs. Also 33 fresh tissues were obtained, fixed and used to detect HPV particles by transitional electron microscopy (TEM). RESULTS: HPV was detected in 32.9% of the tissue specimens by NPCR, in 4.7% by immunohistochemistry and in 28.1% by TEM. In detail, by nested PCR HPV L1 DNA was detected in 40% of normal tissues, 40% of fibromas, 35.8% of non-dysplastic leukoplakias, 31.6% of dysplastic leukoplakias and 22.2% of oral squamous cell carcinomas. The HPV viral load of 96.5% of the samples was very low (1 viral copy per 10(2)-10(4) cells). HPV16 prevails in all histological groups in 89-100%. CONCLUSION: We conclude that HPV does not seem, from the specific sample examined, to play a substantial role in oral carcinogenesis. However, it cannot be excluded that HPV could be involved in oral carcinogenesis only in cases with high viral load or at early stages of carcinogenesis possibly through the hit-and-run mechanism.

VEGF expression from human dysplastic or malignant oral epithelium may be related to mast cell density and the subsequent angiogenetic phenomena.

Vascular endothelial growth factor (VEGF) is an angiogenic cytokine and mast cells play a role in neoangiogenesis in various malignancies. The aim of the present study was to elucidate the role of VEGF and mast cells in the early stages of tumorigenesis in oral squamous cell carcinoma (OSCC). Immunohistochemistry was conducted to study VEGF expression and microvessel density (MVD) in 49 tissue samples, 31 OSCCs, 13 leukoplakias (8 with and 5 without dysplasia) and 5 samples from normal oral tissue. Counterstaining with tolouidine blue was conducted to reveal mast cells. The number of microvessels and mast cells were counted at the same optical field. A gradually increased VEGF expression was observed from normal oral epithelium to leukoplakia and OSCC. MVD was found to increase significantly between normal oral tissue and OSCC (p=0.000). The number of mast cells was found to increase significantly between normal oral tissue, dysplasia (p=0.012) and OSCC (p=0.000). In the early stages of tumorigenesis in OSCC, VEGF, which is secreted by the epithelium, is gradually increased immediately affecting the population of mast cells, which are then related to the increase of microvessels.

The role of reelin gene polymorphisms in the pathogenesis of Alzheimer's disease in a Greek population.

Reelin is an extracellular signaling glycoprotein, which plays a significant role in cytoarchitectonic pattern formation of different brain areas during development. Reelin gene is located on chromosome 7q22. The aim of this study is to investigate the possible association of the following reelin polymorphisms SNP Intron12A/C (rs727531), SNP Exon15A/G (rs2072403), SNP Intron15G/T (rs2072402), SNP Exon22c/g (rs362691), SNP Intron41G/T (rs362719) and SNP Intron59C/T (rs736707) in the pathogenesis of Alzheimer 's disease and the frequency of these polymorphisms in the population of Northern Greece. The study included two groups, A and B. Group A consisted of 50 patients with Alzheimer 's disease and group B of 70 healthy controls. Genomic DNA isolated from blood was used for PCR and subsequent RFLP analysis. According to our results, the exon 22 C/G marker of Reelin is significantly associated with Alzheimer 's disease in the Greek population but the Likelihood Ratio Test shows that the GT haplotype ++ this polymorphism does not affect the phenotype of group A in relation to Group B. This is the first report on a Greek population-based approach.

Isoenzymes of adenosine deaminase and metalloproteinases as biomarkers in in vitro fertilization and embryo transfer.

It is estimated that approximately 1 percent of babies born per year result from in vitro fertilization and embryo transfer, and other assisted reproductive technologies. In humans, the exact mechanisms that lead to embryonic attachment to the endometrial epithelium and invasion into the endometrial stroma have not been fully characterized. The aim of the study is to estimate serum total adenosine deaminase and isoenzymes ADA1, ADA2, as well as MMP-2, MMP-3, MMP-13 and MIP-1a as parameters for pregnancy following IVF-ET. The study group comprised seventeen women who conceived (Group A) and nineteen women aged 21-42 years who did not conceive (Group B) after IVF-ET. Blood samples were collected between 09.00 and 10.00 a.m. during IVF-ET treatment at two different periods. The first blood sample was collected before ET and the second sample 14 days after ET. All serum samples were assayed for the MMP-2, MMP-3 MMP-13 and MIP-1a concentrations with ELISA assay. Serum tADA activity was measured by a spectrophotometer using adenosine as the substrate (Method by Giusti). According to our results it was demonstrated that women who successfully conceived after IVF-ET showed significantly lower serum concentrations of ADA1, MMP-2, MMP-3 and higher serum concentration of MMP-13 at 14 days following ET. In conclusion, ADA1 may play a protective role at the hemochorial interface. Thus, our results suggest that ADA1 may have a modulatory role in the implantation and duration of the pregnancy. In women with successful or unsuccessful pregnancy compared with normal women the levels of ADA and MMPs may be affected by the exogenous hormone therapy according to the protocol of ovarian stimulation during IVF-ET.

HLA alleles and fissured tongue.

The aim of this study was to investigate the linkage between HLA and fissured tongue. Sixty- nine individuals with fissured tongue and 125 healthy volunteers were typed for HLA-DRB1*. The results showed increased frequency of HLA-DRB1*08 (P < 0.001), HLA-DRB1*14 (P < 0.01), HLA-DRB1*11 (P < 0.05) and HLA-DRB1*16 (P < 0.05), while HLA-DRB1*03 and HLA-DRB1*07 frequency was decreased (P < 0.05).

Sarcoidosis: oral and perioral manifestations.

A 46-year-old white woman with lesions on the lower lip, perioral area and in the soft tissues of the oral cavity (gingivae and palate) was examined. The clinical signs were recorded, and incisional biopsies from the oral lesions were taken. The diagnosis of sarcoidosis was established by the histopathological evidence of typical non-caseating granulomas from tissue biopsy, supported by serum ACE- 57.9 U/L, blood calcium 16.83 mEq/L and 24-hrs urine calcium 600 mg). Oral lesions may be the first or the only sign of sarcoidosis in an otherwise healthy patient.

Coexistence of mucous retention cyst and basal cell adenoma arising from the lining epithelium of the cyst. Report of two cases.

OBJECTIVE: To report 2 cases of coexisting mucous retention cyst and basal cell adenoma arising from the lining epithelium of the cyst. CLINICAL PRESENTATION AND INTERVENTION: Two cases of painless swellings, well-demarcated, soft to palpation, and located in the submucosa of the upper lip were clinically examined with the provisional diagnosis of mucocele or salivary gland tumor. Histological examination showed the presence of a large unilocular cystic cavity in many parts surrounded by single or bilayered lining epithelium composed of flattened to cuboidal cells, and in other parts surrounded by projections of cells arranged in a trabecular pattern far into the cystic cavity. The trabeculae were composed of basal and low columnar cells that sometimes formed small duct-like structures. Immunohistochemistry showed that the lining epithelium of the cystic cavity and the cells of the projections expressed cytokeratin 7 and high-molecular-weight cytokeratins. The cells of the projections were weakly positive for S-100 protein and negative for vimentin and alpha-smooth muscle actin. Based on the results, a diagnosis of coexisting mucous retention cysts and basal cell adenomas arising from the lining epithelium of cysts was made. CONCLUSION: The coexistence of mucous retention cysts and basal cell adenomas arising from the lining epithelium of the cyst is reported.

Mast cells and angiogenesis in oral malignant and premalignant lesions.

UNLABELLED: Mast cell contribution to neoangiogenesis during tumorigenesis in oral squamous cell carcinoma is not determined yet. OBJECTIVES: To associate numerical mast cell density (MCD) to numerical microvessel density (MVD) during the progression of oral leukoplakia without dysplasia and leukoplakia with dysplasia to squamous cell carcinoma (OSCC). MATERIALS AND METHODS: MVD was analysed immunohistochemically (mouse monoclonal anti-human CD34) in 49 paraffin-embedded specimens, 35 OSCCs, 9 leukoplakias and 5 normal oral tissues. Toluidine blue counterstaining revealed mast cells. MCD and MVD were assessed at the same optical field. RESULTS: MVD increased between: normal oral mucosa, dysplasia (p=0.004), OSCC (p=0.001), leukoplakia and OSCC (p=0.041). MCD increased between: normal oral mucosa, dysplasia (p=0.003), OSCC (p=0.000), leukoplakia and OSCC (p=0.007). MVD was found to depend on MCD (p=0.000) in a percent 28.3% (power curve fit model). CONCLUSIONS: Mast cells are attracted at the lesion site and may turn on an angiogenic switch during tumorigenesis in OSCC.

Detection of T cells secreting type 1 and type 2 cytokines in the peripheral blood of patients with oral lichen planus.

OBJECTIVE: The purpose of this study was to detect and enumerate T cells secreting type 1 and 2 cytokines in the peripheral blood of patients with oral lichen planus (OLP) and in healthy controls. SUBJECTS AND METHODS: The study group consisted of 80 OLP patients and 80 healthy individuals. Cytokine secreting T cells were detected using ELISPOT assay. RESULTS: There was a statistically significant decrease (p<0.05) in the number of IFN- and IL-12 secreting cells in the peripheral blood of patients with OLP compared to the controls. No statistical difference was observed in the number of IL-2 and TNF-a secreting cells between OLP patients and controls (p>0.05). Also there was no significant difference in the numbers of IFN-gamma, IL-12, IL-2 and TNF-a secreting cells between reticular and erosive forms of OLP (p>0.05). As regards type 2 cytokines, the number of IL-5 and IL-10 secreting cells was significantly decreased in OLP patients compared to the healthy control group (p<0.05). No statistical difference was observed in the number of IL-6 secreting cells between OLP patients and control group (p>0.05). Similarly, no statistical difference was observed in the number of IL-4 secreting cells between OLP patients and controls (p>0.05). No significant difference was also found in the numbers of IL-4, IL-5, IL-10 and IL-6 secreting cells between reticular and erosive OLP group. CONCLUSION: These data suggest decreased type 1 and type 2 cytokine production (except IL-4) in OLP patients.

The role of Candida in inflammatory papillary hyperplasia of the palate.

The objective of this study was to investigate the expression by immunohistochemistry of the major basement membrane (BM) components (laminin, collagen type IV, fibronectin) in specimens from the palatal mucosa lesions of patients with complete dentures and diagnosis of inflammatory papillary hyperplasia of the palate (IPHP). Furthermore to evaluate the potential role of candidal infection in patients with IPHP. Biopsies of palatal mucosa were obtained from patients with IPHP, generally healthy/orally healthy patients with dentures, and healthy subjects. Immunohistochemical studies performed with specific antibodies to BM proteins. Scrapings and swaps of oral lesions from all patients and control groups were taken from the palate, and Candida species colonization was assessed with mycology tests. Immunohistochemical expression of BM components revealed thin linear staining in the BM of healthy palatal mucosa. In IPHP discontinuities or disruptions in BM were observed at the interface between epithelium and the underlying connective tissue in the areas of severe inflammatory response. Our findings suggest an interaction between the expression of BM components and Candida involvement in the development of IPHP, a disorder involving inflammatory reaction and modification of soft tissues.

Verruciform xanthoma coexisting with oral discoid lupus erythematosus.

We describe an unusual case of verruciform xanthoma on the lingual surface of the gum that coexisted with oral discoid lupus erythematosus.

Bilateral aplasia of parotid glands correlated with accessory parotid tissue.

Congenital absence of major salivary glands, especially the parotid gland, is a rare entity. It is usually monolateral and is not correlated with accessory salivary gland tissue. Aplasia of parotid gland may occur alone or in association with abnormalities of other salivary glands, first branchial arch developmental disturbances or other congenital anomalies.We report an interesting case of bilateral aplasia of the parotid glands together with bilateral accessory parotid tissue, without other congenital or developmental anomalies, and we describe the clinical and radiological findings.

Coexistence of lymphoepithelial and epidermoid cysts on the floor of the mouth: report of a case.

Lymphoepithelial and epidermoid cysts in the oral cavity are uncommon. The coexistence of lymphoepithelial and epidermoid cysts in the oral cavity is extremely rare. Only one case of lymphoepithelial cyst associated with two epidermoid cysts on the floor of the mouth has been reported in the literature and the present report describes a second case where a lymphoepithelial cyst coexisted with an epidermoid cyst on the floor of the mouth. It is likely an accidental trauma that was accompanied by inflammation produced the development of implantation-keratinizing epidermoid and lymphoepithelial cysts.

Adenoid cystic carcinoma and polymorphous low-grade adenocarcinoma of minor salivary glands: a comparative immunohistochemical study using the epithelial membrane and carcinoembryonic antibodies.

OBJECTIVE: The purpose of this study was to investigate immunohistochemically the expression of epithelial membrane antigen (EMA) and carcinoembryonic antigen (CEA) in adenoid cystic carcinoma (AdCC) and polymorphous low-grade adenocarcinoma (PLGA) in an attempt to assess the ability of these markers to distinguish AdCC from PLGA when the histological features on routine hematoxylin and eosin are equivocal. MATERIALS AND METHODS: Fourteen specimens of AdCC, 10 PLGA, and five normal minor salivary glands fixed in 10% formalin and embedded in paraffin, were retrieved from the files of our department and were retrospectively studied with the streptavidin-biotin complex method using the epithelial membrane and carcinoembryonic antibodies. RESULTS: The immunoreactivities and the expression patterns of EMA and CEA in AdCC and PLGA were similar. CONCLUSIONS: The results of this study suggest that the immunostaining of AdCC and PLGA with EMA and CEA could not offer an adjunctive aid in differential diagnosis between these two tumors.

Oral Candida isolates in patients undergoing radiotherapy for head and neck cancer: prevalence, azole susceptibility profiles and response to antifungal treatment.

Oral pseudomembranous candidiasis and mucositis were assessed in 39 patients receiving a total dose of 39-70 Gy radiotherapy for head and neck cancer. Mucositis was scored using the Radiation Therapy Oncology Group criteria, and oral candidiasis was diagnosed on the basis of clinical evaluation and quantitative laboratory findings. Radiation-induced mucositis was observed in 9/39 patients. Only 3/39 patients discontinued radiotherapy due to acute severe mucosal effects. Candidiasis (colony-forming units 35 to > or = 60/lesion) associated with mucositis was diagnosed in 30/39 patients: the most frequent aetiology of the infection was Candida albicans (n = 23), followed by Candida glabrata (n = 3), Candida krusei (n = 2), Candida tropicalis (n = 1) and Candida kefyr (n = 1). Patients with confirmed oral pseudomembranous candidiasis were treated with either fluconazole 200 mg/day or itraconazole 200 mg/day for 2 weeks. Clinical improvement and concomitant negative Candida cultures (mycologic cure) were the criteria determining a response to antifungal treatment. Etest revealed very low voriconazole MICs (0.004-0.125 microg/ml) for all isolates, and fluconazole resistance for eight C. albicans strains (MIC > 64 microg/ml) and for the C. krusei isolates (MIC > 32 microg/ml). The same strains showed itraconazole susceptibility dose dependence (MIC 0.5 microg/ml). Despite the itraconazole susceptible dose dependent MIC readings, all patients with oral pseudomembranous candidiasis caused by these strains responded to antifungal treatment with 200 mg/day itraconazole. Oral mycologic surveillance of patients undergoing radiotherapy for head and neck malignancies and susceptibility testing of isolates may be indicated in cases with mucositis-associated confirmed oral pseudomembranous candidiasis to ensure prompt administration of targeted antifungal treatment. On the basis of the low MIC values found, clinical evaluation of voriconazole is indicated for management of oral pseudomembranous candidiasis refractory to other azoles.

Langerhans' cell histiocytosis restricted to the oral mucosa.

Langerhans' cell histiocytosis comprises a group of disorders with Langerhans' cell proliferation as a common feature. The clinical presentation might be highly varied. Typically, there is bone involvement and, less frequently, lesions might be found in other organs, particularly the lungs, liver, lymph nodes, skin, and mucosae. Lesions limited to the oral mucosa are rare. We describe two patients with unifocal eosinophilic granuloma exclusively limited to the oral mucosa. Triamcinolone acetonide infiltration of a palatal lesion yielded a good result in one patient. Six months later, a similar lesion developed in the mandibular gingival mucosa, but it responded to the same treatment. The lesion in the other patient responded to local radiotherapy.

Facial and perioral molluscum contagiosum as a manifestation of HIV infection.

Molluscum contagiosum (MC) is a self-limiting viral disease of the skin and the mucous membranes. Facial and perioral MC is seen with increasing frequency in human immunodeficiency (HIV) infection, particularly in HIV infected homosexual men. The purpose of this study was to describe clinical observations of facial and perioral MC in HIV infected patients. One hundred and eighty HIV-positive individuals (160 males and 20 females) were examined over a period of five years. Fifty-eight were homosexual men and 19 were bisexual men. Fifty-one of 180 patients at the time of the first examination had CD+4 count < 200 cells/mm3 and another 63 presented loss of CD+4 cells in this level, during this period. Three HIV infected patients (two homosexual and one bisexual) were affected with facial and perioral MC. At the time of MC diagnosis the CD+4 count was less than 200 cells/mm3 for all three patients. One patient died nine months after MC diagnosis and the other two are still alive. It is remarkable that in this study no clinical lesions were observed on other sites of the skin.

Proliferating cell nuclear antigen (PCNA) immunostaining in Helicobacter pylori infection: impact of eradication.

Despite the fact that the association of Helicobacter pylori (H. pylori)with an increased risk of gastric cancer has been well documented, the exact mechanisms of this association have not been fully elucidated. The aim of the present prospective study was to contribute to the exploration of these mechanisms by studying the relationship between H. pylori infection and proliferating cell nuclear antigen (PCNA) immunostaining in endoscopic biopsies in gastric antrum. Furthermore, we examined the impact of H. pylorieradication on this relationship. We studied 28 H. pyloripositive patients and the results were compared with 22 endoscopically and histologically normal H. pylorinegative patients (control group) who were comparable to the H. pyloripositive group for age and sex. In addition all H. pyloripositive patients were examined before and after treatment aiming to eradicate H. pylori. In the H. pylori(+) patients the median PCNA index was 35 (range 8-58) and this was significantly higher than the respective number in the control group [5.5 (2 14), p<0.001]. In patients studied before and after successful eradication of H. pylori(n=10) the corresponding numbers were 35 (8-56) and 7 (4 13) (p<0.01) the latter not being significantly different from the control group of H. pylori(-)patients. On the contrary, in patients without successful H. pylori eradication (n=18) the PCNA indices before and after treatment were similar [35.5 (21-58) vs 31.5 (20-56)]. It is concluded that H. pyloriinfection alters the replication cycle of the gastric mucosa inducing hyperproliferation, which return towards normal after successful H. pylori eradication.

Concurrence of torus palatinus with palatal and buccal exostoses: case report and review of the literature.

Exostoses, also known as hyperostoses, are localized bony protuberances that arise from the cortical plate. These developmental anomalies, or hamartomas, frequently affect the skeletal jaw. Different types of exostoses have been described. Torus palatinus and torus mandibularis are two of the most common intraoral exostoses. Other types of exostoses, affecting the palatal aspect of the maxilla (palatal exostoses) or the buccal aspects of the jaws (buccal exostoses), are less commonly encountered. Concurrence of different forms of exostoses in the same individual is very rare. A 48-year-old woman manifesting excessive palatal exostoses, torus palatinus, and buccal exostoses is described. We present the clinical and histopathologic features and applied therapy and provide a comprehensive review of the current features of exostoses.