Impact of combined mycoplasmataceae and HPV co-infection on females with cervical intraepithelial neoplasia and carcinoma.

PURPOSE: The concurrent prevalence investigation of human papillomavirus (HPV), Mycoplasma hominis (Mh) and Ureaplasma urealyticum (Uu) in women in order to estimate the association of co-infection with cervical lesions. METHODS: The study cohort comprised 120 women with no cervical lesions (control group) and 62 women with abnormal cytological findings from the cervix (cervical intraepithelial lesion/neoplasia) as study group. A combination of molecular analyses was implemented. RESULTS: The presence of HPV infection was shown in 52/62 (83.9%) of women with abnormal cytology. Women with cervix cytological findings were shown to have 17.6 times higher risk for Mh and Uu co-infection (p=0.001). HPV and Uu co-infection were detected with a higher prevalence among women with CIN 3 and invasive cancer. CONCLUSION: These findings are consistent with the notion that microbial co-infections may play an important role in persistent inflammation and progression of cervical lesions.

Extraoral surgical removal of an ectopic impacted third molar of the mandible. Report of a case.

Intraoral approach for the removal of impacted third molars represents a common surgical procedure for the specialized clinician. However, in some cases such as ectopic third molars, extraoral surgical removal seems to be inevitable. We present a step by step case of a 56 year old woman with an ectopic third molar of the lower jaw along with a cystic lesion, which were surgically removed by a submandibular approach. Postoperative clinical course was uneventful and there were no signs of facial nerve paresis. In such cases, appropriate preoperative planning must be made based on careful study of radiographic imaging and clinical examination. The more conservative technique that would minimize adjacent anatomic structures risk should be the surgical technique of choice. Key words:Ectopic third molar, mandible, cyst, extraoral approach.

Assessment of TLR4 and TLR9 signaling and correlation with human papillomavirus status and histopathologic parameters in oral tongue squamous cell carcinoma.

OBJECTIVES: Toll-like receptors (TLRs) may promote or inhibit tumor progression. The aim of this study was to assess the expression of TLR4 and TLR9 and their downstream targets in oral tongue squamous cell carcinoma (OTSCC) in correlation with histopathologic parameters and human papillomavirus (HPV) status. STUDY DESIGN: OTSCC (fully or superficially invasive and in situ) were studied. Immunohistochemical expression of TLR4, TLR9, nuclear factor-κΒ (NF-κΒ/p65), and interferon-ß (IFN-ß) was evaluated in tumor and inflammatory cells and in adjacent morphologically normal mucosa. HPV status was also determined. RESULTS: TLR4 showed increased expression levels in tumor and infiltrating inflammatory cells compared with adjacent mucosa, especially in fully invasive cases; a negative correlation between TLR4 levels in inflammatory cells and tumor grade was observed. TLR9 was upregulated in tumor and infiltrating inflammatory cells compared with the adjacent mucosa; its expression in inflammatory cells was higher in well differentiated tumors. NF-κΒ and IFN-ß were elevated in cancerous tissues, especially in fully invasive cases, and positively correlated with TLR4 and/or TLR9. HPV positivity (detected in 15.9% of the cases) demonstrated positive correlation with TLR9 and NF-κΒ levels. CONCLUSIONS: TLR4 and TLR9 are upregulated in OTSCC and its microenvironment and, by affecting important downstream molecules, such as NF-κB and IFN-ß, may play a role in oral cancer development and progression.

Expression of NF-κB and IL-6 in oral precancerous and cancerous lesions: An immunohistochemical study.

BACKGROUND: The purpose of this study was to evaluate the immunohistochemical expression of NF-κB and IL-6 in oral premalignant and malignant lesions and to investigate their possible correlation with the presence of subepithelial inflammation. MATERIAL AND METHODS: Thirty two oral premalignant lesions, clinically compatible with leukoplakia or erythroplakia, were investigated. Microscopically, 11 of them showed hyperkeratosis and acanthosis (epithelial hyperplasia) and 21 showed dysplasia of varying degrees. Nine cases of OSCC and four control cases of normal oral mucosa were also included in the study. Immunohistochemical staining with NF-κB (p65) and IL-6 was performed. IL-6 and nuclear NF-κB staining were assessed as positive or negative. For cytoplasmic localization of NF-κB, a total score combining intensity and percentage of positive epithelial cells was additionally calculated. The presence of inflammation was also recorded. RESULTS: Intensity and total scores for NF-κΒ cytoplasmic immunostaining showed a statistically significant gradual increase from normal mucosa to OSCC (p=0.012 and p=0.026 respectively). Non-statistically significant increased NF-κΒ nuclear localization was detected in dysplasias and OSCCs. Positive statistical correlation was detected between the presence of inflammation and IL-6 expression (p=0.015). No correlation between NF-κΒ and IL-6 was detected. CONCLUSIONS: NF-κΒ is activated in the early stages of oral carcinogenesis. IL-6 may have an NF-κΒ-independent role, possibly through regulation of the inflammatory response.

Phosphorylated signal transducer and activator of transcription-1 immunohistochemical expression is associated with improved survival in patients with oral squamous cell carcinoma.

PURPOSE: To estimate whether the immunohistochemical (IHC) expression patterns of the tumor suppressor gene signal transducer and activator of transcription-1 (STAT1) and its active phosphorylated form (PSTAT1) serve as potential prognostic and predictive markers in patients with oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: STAT1 and PSTAT1 protein expressions were examined immunohistochemically in OSCC tumor tissues and adjacent normal mucosa from 49 patients who underwent primary surgery. The IHC scores were correlated with all available clinicopathologic parameters that were obtained from a maximum of 7 years of follow-up, including survival and response to adjuvant therapy treatment. RESULTS: There was a shift toward lower percentages of cells with STAT1 (P < .014) and PSTAT1 (P < .001) detected in OSCC tumors compared with adjacent normal tissue sites. No association with patients' clinicopathologic characteristics was shown. However, for the group of patients who received adjuvant chemotherapy, increased PSTAT1 intensity of staining in OSCC tumors was strongly associated with better overall survival (P = .008). CONCLUSIONS: This is the first study to concurrently evaluate STAT1 and PSTAT1 IHC expression patterns and their prognostic significance in patients with OSCC, highlighting the potential role of PSTAT1 as a biomarker in therapeutic decision making. Large prospective studies are needed to verify these findings.

Effect of thrombosis-related gene polymorphisms upon oral cancer: a regression analysis.

It is well-known that there is an interplay between hemostasis, thrombosis and cancer. Functional DNA polymorphisms in genes encoding factors related to thrombosis have been associated with increased risk for oral squamous cell carcinoma (OSCC). The present study investigated the possible combinatory effect of 10 such polymorphisms as primary risk predictors for OSCC in a European population. Two groups including 160 patients with OSCC and 168 healthy controls of Greek and German origin were studied. The patient and control groups were comparable regarding ethnicity, age and gender. For all studied individuals, 10 genotypes of functional polymorphisms were investigated: 5,10-methylene tetrahydrofolate reductase (MTHFR) C677T, coagulation factor V (F5) Leiden, coagulation factor II (F2, also known as prothrombin) G20210A, coagulation factor XII (F12) C46T, coagulation factor XIII A1 subunit (F13A1) Val34Leu, serpine1 (SERPINE1, also known as plasminogen activator inhibitor-1) 4G/5G, protein Z (PROZ) -A13G, angiotensin I-converting enzyme (ACE) I/D, angiotensinogen (AGT) Met325Thr, and carboxypeptidase B2 (CPB2, also known as thrombin-activatable fibrinolysis inhibitor) C1040T. Multivariate logistic regression models were used in order to evaluate the relation and contribution of homozygous and heterozygous variant polymorphisms upon overall, early and advanced stages of OSCC. Five out of the studied polymorphisms, influencing the expression of SERPINE1 and ACE genes, as well as the activity of CPB2, F12 and F13 proteins, were recognized as significant predictive factors for OSCC. The 'mode of inheritance' regression model, in particular, revealed the low expression I allele of ACE to be a primary predictor in overall, early and advanced stages of oral cancer. Comparing the present findings with previous knowledge, possible interactions of these factors and their relation to the risk for OSCC development are discussed.

Prevalence of human papillomavirus in saliva and cervix of sexually active women.

OBJECTIVE: Human papillomavirus (HPV) is strongly associated with cervical cancer and possibly with some oropharyngeal cancers. However, the relation between oral and cervical HPV infection is not fully understood. This study evaluates the prevalence rate and type-concordance of HPVs in these areas. METHODS: HPV DNA typing was performed in saliva and cervical specimens of 43 sexually active women, with the use of general consensus PCR and nested PCR (NPCR) tests. RESULTS: The prevalence rate of HPV DNA in cervical and saliva samples was 51.2% and 11.6% with general PCR, and 60.5% and 44.2% with NPCR, respectively. The probability of HPV DNA detection with general PCR in saliva was about 8 times lower compared to the cervix (P<0.001, OR: 0.13, 95% CI: 0.04-0.37), but showed no difference when the more sensitive NPCR method was applied (P=0.139, OR: 0.52, 95% CI: 0.22-1.21). The distribution of HPV variants according to their oncogenic potential revealed no statistically significant difference, regardless to the PCR method used for both sites. All general PCR HPV DNA positive saliva specimens belonged to women with cytology findings (n=5). These women had also 8.5 times higher risk for presenting with positive HPV detection in saliva with the NPCR method (P=0.009, OR=8.50, 95% CI: 1.74-39.70). CONCLUSIONS: Women with genital HPV infection are at higher risk for asymptomatic oral HPV infection. Prophylactic HPV-vaccination may reduce the burden of HPV-related diseases in both cervix and oropharynx.

Association of polymorphisms in Tumor Necrosis Factor Alpha and Beta genes with increased risk for oral cancer.

BACKGROUND: In light of the recently found contribution of inflammation-related factors to oral oncogenesis, the possible correlation of tumor necrosis factor alpha and beta genes (TNF-alpha and TNF-beta) with risk of oral cancer was investigated. MATERIALS AND METHODS: DNA samples of 160 German and Greek patients with oral squamous cell carcinoma and 153 healthy controls of equivalent age, gender and ethnicity were studied. The functional polymorphisms TNF-alpha (-308 G/A) and TNF-beta (252 G/A), which affect gene expression, were investigated by restriction fragment length polymorphism analysis. RESULTS: The frequencies of high expression A2 (-308A) TNF-alpha allele and high expression B1 (252G) TNF-beta allele were significantly increased in cancer patients compared to controls (respectively: 62.2% versus 14.7%, p<0.0001; OR 8.65, 95% CI 5.74-13.04 and 66.9% versus 15.7%, p<0.0001; OR 10.92, 95% CI 7.4-16.2). Three combined TNF-alpha/TNF-beta genotypes (A2A2/B1B1, A1A2/B1B2, A1A2/B1B1) were over-represented in cancer patients (p<0.001). No significant differences in allele frequencies were detected among most subgroups of patients divided in regard to cancer stage, family history for cancer or thrombosis, smoking or heavy alcohol consumption habits. CONCLUSION: This study showed a strong association of TNF-alpha and TNF-beta high expression alleles with increased risk of oral cancer. These findings are in accordance with previously observed high TNF-alpha levels in serum of patients with oral cancer in comparison to healthy controls.

Association of leptin -2548G/A and leptin receptor Q223R polymorphisms with increased risk for oral cancer.

PURPOSE: We investigated the possible association of DNA polymorphisms -2548G/A and Q223R in the leptin (LEP) and leptin receptor (LEPR) genes, respectively, which both affect the amount of circulating cytokine-type hormone leptin, with risk for development of oral cancer. METHODS: Polymerase chain reaction-based restriction analysis was performed in DNA samples of 150 patients with oral squamous cell carcinoma (OSCC) and 152 healthy control subjects of equivalent gender, age, and ethnicity (Greeks and Germans). RESULTS: Compared to controls, the homozygous high gene expression genotype A/A of the LEP -2548G/A polymorphism was significantly increased in the subgroups of patients with advanced cancer stages (P = 0.0001; OR 9.0, 95% CI 2.62-30.89), with a positive family history of cancer (P = 0.0346; OR 3.55, 95% CI 1.15-11.01), without tobacco abuse (P = 0.0051; OR 9.69, 95% CI 1.03-91.24), and without alcohol abuse (P = 0.0472; OR 2.16, 95% CI 0.87-5.37). The homozygous low-leptin-binding genotype G/G of the LEPR Q223R polymorphism was strongly associated with an increased risk for OSCC for all patients (P = 0.0028; OR 4.11, 95% CI 1.30-12.97) as well for most of the patient subgroups. CONCLUSIONS: The above findings are consistent with the growth-promoting role of leptin in cancer and its induction effect on angiogenesis and metastasis. This is the first study indicating the association of these LEP and LEPR gene polymorphisms with increased risk for OSCC.

Orbital cellulitis, orbital subperiosteal and intraorbital abscess: report of three cases and review of the literature.

INTRODUCTION: Orbital cellulitis is usually a complication of paranasal sinus infection. Either the infection may dissect under the periosteum and lead to subperiosteal abscess (SPA) or intraorbital abscess may be formed secondary to a progressive and localized cellulitis. Without appropriate treatment orbital infection may lead to serious complications, even death. REPORT OF CASES: Three cases are described, one of orbital cellulitis, one of SPA and one of intraorbital abscess and the literature is being reviewed. CONCLUSION: Prompt treatment is mandatory to avoid visual loss or intracranial complications. Initially, IV antibiotics may be administered, but if no improvement appears within 48h, surgical drainage of the orbit and the affected sinuses must be performed. In medial or medial-inferior SPA a transnasal approach is used, but in superior orbital abscess an external incision is required.

Angiotensinogen polymorphism is associated with risk for malignancy but not for oral cancer.

BACKGROUND: In light of the recently found contribution of angiogenic and inflammation-related factors to malignancies, this study investigated the possible association of the angiotensinogen gene (AGT) with increased risk of oral cancer. MATERIALS AND METHODS: The M235T polymorphism, which influences AGT gene expression, was evaluated by restriction fragment length polymorphism analysis in the DNA samples of 163 German and Greek patients with oral squamous cell carcinoma (OSCC) and 124 healthy controls of equivalent gender, ethnicity and age. RESULTS: No significant difference of the mutant (235T) allele, which results in higher AGT gene expression, was observed in the whole patient group in comparison with the normal controls. Similarly, compared to the controls no significant difference of either allele or carrier frequency was detected in almost every subgroup of patients. Only in the subgroup of patients with a positive family history of cancer was a significant increase of mutant T allele and carrier frequencies observed, compared to the controls (50% vs. 36.7% and 79.3% vs. 61.3%, respectively, p < 0.05 in both cases). In this particular subgroup of patients the odds ratio for OSCC of TT homozygotes was 3.57 (CI 95% 1.2-10.62), while for the MT heterozygotes it was 2.41 (CI 95% 1.06-5.49). CONCLUSION: This study did not reveal an association of the AGT M235T polymorphism with oral oncogenesis, but certainly suggested a possible association of this specific polymorphism with other types of cancer. The present findings support a previous suggestion that the pathway of oral oncogenesis is probably based on angiotensin-converting enzyme and bradykinin interaction and not on AGT and angiotensin peptides.

Delayed appearance of diplopia due to orbital emphysema after repair of orbital fractures.

A 38-year-old caucasian male, after continuous nose blowing caused by common flu, developed left diplopia, especially in downward gaze, and complained of orbital dull pain during the preceding 48 hours. Clinical signs included left lower eyelid crepitant edema, mild exophthalmos and restriction of the eye movement. The patient had been operated on for orbital fractures 12 days previously. Open reduction had been performed, stabilization had been achieved with osteosynthesis miniplates, and the orbital floor defect was reconstructed with a piece of Liodura. Orbital computerized tomographic scan demonstrated a large air collection, diagnostic of emphysema, without displacement of the fractures. Under local anesthesia, aspiration-decompression was performed using a 25-gauge needle. On the next day, the patient was free of pain, and 2 days after surgery the diplopia and exophthalmos resolved with no sequelae.

Gene expression polymorphisms of interleukins-1 beta, -4, -6, -8, -10, and tumor necrosis factors-alpha, -beta: regression analysis of their effect upon oral squamous cell carcinoma.

PURPOSE: Functional DNA polymorphisms affecting gene expression and serum or saliva levels of interleukins IL-1 beta,-4,-6,-8,-10 and tumor necrosis factors TNF-alpha,-beta have been associated with increased risk for the development of oral squamous cell carcinoma (OSCC). The present retrospective case-control study examines possible interactions between seven cytokine genotype polymorphisms and their combinatory effect in predicting the occurrence of OSCC in Caucasians. METHODS: Three hundred and thirty Greeks and Germans were studied, consisting of 162 OSCC cases and 168 healthy controls of comparable age, gender, and ethnicity. A series of multivariate logistic regression models, adjusted for age and gender, was constructed in order to assess the contribution of homozygous or heterozygous variant genotypes of polymorphisms IL-1 beta (+3953C/T), IL-4 (-590C/T), IL-6 (-174G/C), IL-8 (-251A/T), IL-10 (-1082A/G), TNF-alpha (-308G/A) and TNF-beta (+252G/A) upon overall, early and advanced stages of OSCC development. RESULTS: The contribution of TNF-alpha and IL-6 was consistent and robust in almost all models constructed. Furthermore, when the mode of inheritance of each variant allele was taken into account in a "biological" multivariate logistic regression model, four polymorphisms emerged as primary predictors for overall stages of OSCC: TNF-alpha (OR = 15.27; 95% CI = 7.30-31.96), IL-6 (OR = 8.33; 95% CI = 3.95-17.58), IL-8 (OR = 3.54; 95% CI = 1.69-7.43) and IL-10 (OR = 2.65; 95% CI = 1.28-5.46). Finally, IL-1 beta, IL-4 and TNF-beta polymorphisms were not primary predictors of OSCC development in all constructed models. CONCLUSIONS: This study revealed the highly significant contributions of two out of seven studied cytokines (IL-6 and TNF-alpha) in the occurrence of OSCC. Based on these findings and previous reports, possible stoichiometrical interactions of cytokines leading to OSCC development are discussed.

High gene expression of matrix metalloproteinase-7 is associated with early stages of oral cancer.

BACKGROUND: In the light of the recently found contribution of matrix metalloproteinases (MMPs) to oral oncogenesis, the correlation of MMP-7 with risk for oral cancer was investigated. MATERIALS AND METHODS: The MMP-7 -181A/G polymorphism in 159 German and Greek patients with oral squamous cell carcinoma and 120 healthy controls of equivalent gender and ethnicity was studied. RESULTS: The detected carrier frequency of the high expression G allele was significantly higher in patients compared to controls (74.8% versus 61.7%, p = 0.0257). This significant difference was more pronounced in patients with early stages of cancer and absent in those with advanced stages. A/G heterozygotes have a double relative risk (OR 2.07, 95%, CI 1.17-3.67) of developing early stages of oral cancer than low expression A/A homozygotes. CONCLUSION: MMP-7 gene expression is associated with increased risk only for early stages of oral cancer, possibly due to the inhibitory effect of MMP-7 in angiogenesis.

Strong association of interleukin-4 (-590 C/T) polymorphism with increased risk for oral squamous cell carcinoma in Europeans.

OBJECTIVE: The present study was performed to investigate the possible association of -590 C/T polymorphism in the interleukin-4 (IL-4) gene which affects its expression with the risk for development of oral cancer. STUDY DESIGN: Polymerase chain reaction-based restriction analysis was performed in DNA samples of 156 patients with oral squamous cell carcinoma (OSCC) and 162 healthy control subjects of equivalent gender, age, and ethnicity (Greek and German). Statistical analyses were performed conducting Fisher exact test. RESULTS: The T/T genotype was associated with an increased risk for the development of OSCC (P = .018; OR 1.47, 95% CI 0.66-3.28), especially for early stages of this malignancy (P < .0001; OR 3.17, 95% CI 1.31-7.65). CONCLUSIONS: The above findings are consistent with the growth-promoting role of IL-4 in head and neck cancer and its inhibitory effect on neoangiogenesis and metastasis. The present study in Europeans is not in accordance with a previous report of unclear association of this polymorphism in a Chinese population.

Strong association of the tissue inhibitor of metalloproteinase-2 polymorphism with an increased risk of oral squamous cell carcinoma in Europeans.

The present study was performed in order to investigate the possible association of the -418 G/C polymorphism in the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene, which affects its expression, with the risk of developing oral cancer. PCR-based restriction analysis was performed in DNA samples from 158 patients with oral squamous cell carcinoma (OSCC) and 168 healthy controls of equivalent sex, age and ethnicity (Greeks and Germans). Statistical analyses were performed with Fisher's exact test and the calculation of odds ratios with a 95% confidence interval (CI). The frequency of the low C allele expression was ten times greater in the patients than the controls (31% vs 2.7%, respectively; P<0.001). The C/C and G/C genotypes were associated with an increased risk of developing OSCC (P<0.001, OR=40.88, 95% CI=2.24-744.40, and P<0.001, OR=21.31, 95%=9.82-46.21, respectively). The same pattern of significant differences with the controls was also observed in the subgroups of patients in regard to the initial or advanced stages of oral cancer, family history of any type of cancer or thrombosis, and smoking habits or alcohol abuse. These findings are consistent with the reduced levels of TIMP-2 in the presence of the low expression C allele, which are insufficient to inhibit the matrix metalloproteinase-driven degradation of the extracellular matrix (leading to cancer invasion) and mitogen-driven neoangiogenesis (leading to tumor growth and metastasis). In conclusion, the studied TIMP-2 polymorphism is strongly associated with an increased risk of OSCC in Europeans carrying the low C allele expression. These results indicate that this polymorphism could serve as a genetic marker for the susceptibility of cancer in the oral cavity.