Coinfection with Epstein-Barr Virus (EBV), Human Papilloma Virus (HPV) and Polyoma BK Virus (BKPyV) in Laryngeal, Oropharyngeal and Oral Cavity Cancer.

Most research providing evidence for the role of oncogenic viruses in head and neck squamous cell carcinoma (SCC) development is focused on one type of virus without analyzing possible interactions between two or more types of viruses. The aim of this study was to analyse the prevalence of co-infection with human papillomavirus (HPV), Epstein-Barr virus (EBV) and polyoma BK virus (BKPyV) in oral, oropharyngeal and laryngeal squamous cell carcinomas in Polish patients. The correlations between viral infection, SCC, demographic parameters, evidence of metastases and grading were also investigated. Fresh-frozen tumour tissue samples were collected from 146 patients with laryngeal, oropharyngeal and oral cancer. After DNA extraction, the DNA of the studied viruses was detected using polymerase chain rection (PCR) assay. Males (87.7%) with a history of smoking (70.6%) and alcohol abuse (59.6%) prevailed in the studied group. Histological type G2 was recognized in 64.4% cases. The patients were most frequently diagnosed with T2 stage (36.3%) and with N1 stage (45.8%). Infection with at least two viruses was detected in 56.2% of patients. In this group, co-infection with HPV/EBV was identified in 34.1% of cases, EBV/BKV in 23.2%, HPV/BKV in 22.0%, and HPV/EBV/BKV in 20.7%. No difference of multiple infection in different locations of cancer was observed. The prevalence of poorly differentiated tumours (G3) was more frequent in co-infection with all three viruses than EBV or BKV alone. A significant correlation was observed between tumour dimensions (T) and lymph-node involvement (N) in co-infected patients compared to single infection. Further studies are necessary to clarify whether co-infection plays an important role in the initiation and/or progression of oncogenic transformation of oral, oropharyngeal and laryngeal epithelial cells.

Lipids and lipoproteins and inflammatory markers in patients with chronic apical periodontitis.

BACKGROUND: Since chronic apical periodontitis (CAP) appears to be a risk factor for coronary heart disease, the aim of the study was to determine the relationship between the size of CAP lesion and inflammatory markers (hsCRP, IL-6, TNF-α), as well as lipids and lipoproteins (LpPLA2, apoAI, apoB level) in blood serum of patients with CAP. METHODS: The patients studied (n = 43) were divided into groups: patients under 50 and over 50 years of age, and a separate subgroup of the oldest age with the largest size of CAP lesions. Apolipoprotein AI (apoAI) above 150 mg/dL and below 150 mg/dL was used as an important criterion for the division of patients into groups. The CAP lesion size was measured using the Kodak digital imaging system software. The control group consisted of clinically healthy volunteers (n = 20) without CAP. Lipids were measured on a Siemens analyzer (Germany), apoAI, apoB, hsCRP levels were determined by immunonephelometric method, using the Health Care Diagnostic Product (Siemens GmbH, Germany), and IL-6, TNF-α and LpPLAG7 assay kits (ELISA, R&D Systems) were used. RESULTS: The findings suggested that in patients with CAP and their age increase, the CAP lesion size, the concentration of inflammatory markers and LpPLA2 mass increased. Correlations between the CAP lesion size and LpPLA2 mass and between the CAP lesion size and TG level in patients with apoAI 150 ≤ mg/dL showed increase TG in atherogenic apoB-containing triglyceride-rich lipoprotein and TC in cholesterol-rich lipoprotein. The patients with a low apoAI and high LpPLA2 level can have a higher risk of odontogenic disease and progression of atherosclerosis and coronary heart disease. CONCLUSION: We have found a positive correlation between apoAI level and the CAP lesion size and a negative correlation between LpPLA2 level and the CAP lesion size. The results suggest that apoAI and LpPLA2 in HDL particles have antiinflammatory action and together can limit the CAP lesion size in patient with a higher apoAI level. The literature data on the distribution of lipoprotein particles in subjects are still insufficient, so this problem requires further studies.

[Television and Internet as sources of women knowledge of tobacco smoking, alcohol consumption and energy drinks impact on health]. / Telewizja i internet jako zródla wiedzy kobiet o wplywie palenia papierosów, spozywania alkoholu i napojów energetyzujacych na stan zdrowia.

INTRODUCTION: Accessibility of the Internet allows obtaining information on different areas of life, including the impact of smoking, alcohol consumption and energy drinks on health. Environmental exposure to tobacco smoke and active smoking are a serious risk for women's health, especially for women in reproductive age and children at any time in their lives. Alcohol is a risk factor for the development of general diseases, and consumed by pregnant women has a toxic effect on the body of women and a child in the prenatal period. Due to the increased consumption of energy drinks containing among others nervous system stimulants and carbohydrates, their consumption should be a conscious choice of the consumers. Knowledge of the health risks resulting from the lifestyle can be a decisive factor for the implementation of health behaviour. The aim of the study was to determine the sources from which men and women acquire information concerning the effects of cigarette smoking, alcohol consumption and energy drinks on health. The respondents interest in the above mentioned subjects was also evaluated. MATERIAL AND METHODS: The survey study was carried out in a group of 160 persons (114 women and 46 men), aged 19-60 years, randomly selected from the patients presenting to the Department of Conservative Dentistry with Endodontics of the Medical University of Lublin. An author's questionnaire was prepared for this research. The data were analyzed statistically with the use of Pearson's X2 test. Statistically significant test values were those with p<0.05. RESULTS: The internet was a source of information about the impact of smoking cigarettes on health for 52.63% women and 56.52% men, about the alcohol effect on health for 57.02% women and 45.65% men, while about energy drinks for 61.40 % of women and 47.83% men. Differences between sex of the respondents and indicated source of information were not statistically significant. Obtaining information from television programmes on the impact of smoking on health reported 70.18% of women and 63.04% of men, about alcohol consumption - 66.67% women and 58.70% men respectively. There was no statistically significant correlation between sex of the respondents and obtaining information concerning cigarette smoking and alcohol consumption. Women showed significantly more frequently TV programmes as a source of information about energy drinks (61.40%) compared to males (43.48%) (X2 = 4.28, p <0.05). Interest in the subject of the impact of smoking, alcohol consumption and energy drinks on health was not dependent on sex of the respondents. CONCLUSIONS: Most women acquire knowledge about the effects of cigarette smoking, alcohol consumption and energy drinks on health from TV programmes, although the internet is an increasingly important source of information in this respect.

[Chewing gum as an additional agent in maintaining oral hygiene versus smoking status--preliminary study]. / Guma do zucia jako srodek dodatkowy w utrzymaniu higieny jamy ustnej a status palenia--badanie wstepne.

Nowadays chewing gum is widely used in different age groups, so complying with proper duration and frequency of chewing is an important factor influencing the state of masticatory system. The study involved 112 dental students of the Medical University of Lublin. Everyday use of chewing gum declared 47,32% of cases. Chewing time up to 10 minutes was stated in 23,08% of respondents, 11-20 minutes in 40,38% of interviewees. Among the examined students 17,3% smoked cigarettes. In smokers group 83,33% of questioned chewed the gum every day, while among non-smokers - 43,37%. Chewing time shorter than 10 minutes declared 22,22% of smokers and 23,26% of non-smokers, while chewing time between 11-20 minutes - 27,78% i 44,35% of smokers and non-smokers respectively. Obtained results indicate the need of carrying out further studies aimed at the nicotine influence on saliva parameters with respect to development of diseases of hard tooth tissues.

Serum and Tissue Level of TLR9 in EBV-Associated Oropharyngeal Cancer.

The Epstein-Barr virus (EBV) is associated with the development of various epithelial malignancies including cancer in the head and neck region. Several studies have shown that Toll-like receptors (TLRs) are required for an innate immune response to infection with human DNA viruses, e.g., EBV. During viral infections, TLR response may influence the transformation to malignancy. The aim of the study was to assess TLR9 serum and tissue level in EBV(+) and EBV(-) oropharyngeal cancer patients. The study involved 78 patients: 42 EBV(+) and 36 EBV(-). EBV DNA was detected in fresh frozen tumor tissue. TLR9 level was measured in homogenate of tumor tissue and in serum. Moreover, in serum samples IL-10, VEGF, TGFß, TNFα and antibodies against EBV were detected using ELISA test. TLR9 level was significantly lower in EBV(+) patients, both in tissue and serum, while EBVCA, EBNA and VEGF level was statistically higher in EBV(+) patients. An increase in EBVCA and EBNA antibodies titer was correlated with a TLR9 level decrease. TLR9 level was higher in poorly-differentiated tumors (G3), in tumor of larger dimensions (T3-T4) and with lymph nodes involvement (N3-N4) but without statistical significance. High levels of anti-EA antibodies in the majority of EBV(+) patients may point to the reactivation of EBV infection.

[Tobacco smoking-related attitudes of dental students of the Medical University of Lublin]. / Postawy studentów stomatologii Uniwersytetu Medycznego w lublinie wobec problemu palenia tytoniu.

Despite the growing consciousness of negative tobacco smoking consequences there is still a need of propagating healthy lifestyle among the society. Physicians and dentists as well as medical and dental students have a particular role in this process due to their daily contact with smokers. The aim of the study was evaluation of tobacco smoking prevalence among dental students of the Medical University of Lublin as well as recognizing their knowledge and attitudes regarding negative tobacco smoking consequences. The questionnaire survey was carried out among 112 dentistry students, tobacco smoking was reported in 16.07%. The students smoked most frequently 5-10 cigarettes per day for 4.3 years. 44.44% of students started smoking during their studies. The main reason of smoking was relaxation and pleasure or in respond to stressful situations. The half of the respondents made one or more attempts of quitting smoking with different results. The most often potential cause of smoking cessation was pregnancy. Almost 75% of students could not indicate any tobacco dependent disease or they could indicate only one - lung cancer.

[Oral hygiene habits among tobacco-smoking and non-smoking students of the Medical University of Lublin--chosen aspects]. / Nawyki dotyczace higieny jamy ustnej wsród palacych tyton i niepalacych studentów Uniwersytetu Medycznego w Lublinie--wybrane aspekty.

Among several etiologic factors for dental caries and periodontal diseases we can find dental plaque that forms on the teeth surfaces and prosthetic appliances. Elimination of dental plaque by proper oral hygiene procedures is crucial in caries and periodontal disease prevention. The aim of the study was evaluation of tobacco smoking prevalence among dental students of the Medical University of Lublin and the comparative analysis of oral hygiene habits among smoking and nonsmoking students. A questionnaire survey was carried out among 112 students of the Medical University of Lublin during the second, third, fourth and fifth year of their studies. The students were 20-28 years of age. The questions concerned cigarettes smoking habit and the ways of maintaining oral hygiene. Respondents were divided into smoking and non-smoking group. Statistical analysis was carried out. Obtained results were sent to statistical analysis. Cigarette smoking was reported by 16.67% of surveyed students. No significant differences between smoking and non-smoking students were stated in frequency of brushing, changing the toothbrush, density of toothbrush filaments, using manual and power toothbrush, using whitening toothpastes and frequency of using dental floss and toothpicks. Statistically significant difference was noted in gum chewing habit--smoking students chewed the gum more frequently (83.33%) than non-smoking students (40%). Significant differences occurred also in frequency of professional removal of dental deposits. Calculus removal performed twice a year was reported by 50% of smoking students, comparing with 17.8% of nonsmoking students. 37.78% of nonsmoking students declared professional teeth cleaning performed more often than twice a year comparing with 11.11% of respondents from the smokers group (p < 0.05).

Glutathione Peroxidase (GPx) and Superoxide Dismutase (SOD) in Oropharyngeal Cancer Associated with EBV and HPV Coinfection.

Recent reports have pointed to the link between persistent inflammation, oxidative stress, and carcinogenesis; however most of the studies concerning the role of viruses in head and neck cancer (HNC) are focused mainly on one type of virus. Our present study aimed to study the relationship between Epstein-Barr virus/human papilloma virus (EBV/HPV) coinfection and glutathione peroxidase (GPx) and superoxide dismutase (SOD) level in oropharyngeal cancer. Fresh-frozen tumor tissue samples were collected from 128 patients with oropharyngeal cancer infected with EBV or HPV or with EBV/HPV coinfection. After DNA extraction, EBV and HPV DNA was detected using a polymerase chain reaction (PCR) assay. GPx and SOD activity was determined in homogenates of cancer tissue using diagnostic kits produced by Randox Laboratories. Both GPx and SOD activity was statistically lower in patients with EBV/HPV coinfection than in a single EBV or HPV infection. Analysis of GPx and SOD activity in relation to histological grading and tumor, node (TN) classification revealed that in poorly-differentiated tumors, the level of antioxidant enzymes was lower compared with well-differentiated lesions and in cases with greater tumor dimensions and lymph-node involvement, both GPx and SOD activity was decreased. Further studies are necessary to clarify the influence of interplay between EBV, HPV, and oxidative stress on malignant transformation of upper aerodigestive tract epithelial cells.

Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity in patients with diabetes mellitus type 2 infected with Epstein-Barr virus.

Oxidative stress is suggested to be the crucial factor in diabetes mellitus type 2 (DM2) pathogenesis and in the development of diabetic complications. Patients with DM2 may be more susceptible to infections due to hyperglycaemia-induced virulence of various microorganisms. Several studies pointed that Epstein-Barr virus (EBV) infection is associated with reactive oxygen species (ROS) production and/or activation of signalling pathways connected with ROS. The present study analyzed serum activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD) in DM2 patients with and without EBV infection. Blood and saliva were collected from 120 patients with DM2. EBV DNA was detected in the saliva using nested-PCR technique. Spectrophotometric methods were implemented to determine serum GPx and SOD activity with the use of diagnostic kits produced by Randox Laboratories. GPx and SOD activity was decreased in diabetic patients, with the lowest values in DM2 EBV-positive patients. There was correlation between GPx and SOD activity-with increased value of GPx, SOD activity was also rised. In patients with DM2 history longer than 10 years as well as in DM2 patients with obesity, antioxidant enzymes activity was decreased. Determination of examined parameters may be useful in diabetic patients with EBV infection and could be important prognostic factor.

[Cigarette smoking and alcohol consumption in patients with oral cancer]. / Palenie papierosów i spozywanie alkoholu w grupie chorych z nowotworami zlosliwymi jamy ustnej.

The study aim was to analyse the frequency of simultaneous smoking and alcohol consumption in 142 patients with oral cancer and in the control group. A connection between the prevalence of these habits and epidemiological traits of the patients and features of malignant lesions was evaluated. Statistical analysis revealed that simultaneous smoking and alcohol abuse were significantly more frequent in the cases group. A strong influence of these habits on the development of oral squamous cell carcinoma was noted.

Prevalence of Epstein-Barr virus, human papillomavirus, cytomegalovirus and herpes simplex virus type 1 in patients with diabetes mellitus type 2 in south-eastern Poland.

A microbiota is a complex ecosystem of microorganisms consisting of bacteria, viruses, protozoa, and fungi living in different niches of the human body, which plays an essential role in many metabolic functions. Modifications in the microbiota composition can lead to several diseases, including metabolic disorders. The aim of this study was to analyze the prevalence of four viruses which can cause persistent infections-Epstein-Barr virus (EBV), human papillomavirus (HPV), cytomegalovirus (CMV), and herpes simplex virus type 1 (HSV-1) in patients with diabetes mellitus type 2 (DM2). Blood, saliva and oral swabs were collected from all the study participants. The nested-PCR technique was used to detect the viral DNA. DNA of at least one virus was detected in 71.1% of diabetic patients and in 30% of individuals without diabetes. In patients with diabetes EBV DNA was detected the most frequently (25.4%), followed by HPV- 19.1%, HSV- 10.4% and CMV- 5.2%. A higher percentage of EBV+HPV co-infection was found among men (30.8%). EBV DNA was statistically more often detected in patients living in rural areas (53.7%), while HPV (91.5%) and EBV+HPV co-infection (22.2%) prevailed among patients from urban areas. In patients with a DM2 history longer than 10 years viral infection was detected more frequently. The prevalence of EBV, HPV and the EBV+HPV co-infection was significantly higher in diabetic patients than in individuals without diabetes. The frequency of these infections depended on the duration of the disease (DM2).

Total Antioxidant Status (TAS), Superoxide Dismutase (SOD), and Glutathione Peroxidase (GPx) in Oropharyngeal Cancer Associated with EBV Infection.

A growing number of studies reveal that oxidative stress is associated with viral infections or cancer development. However, there are few reports assessing the relationships between oxidative stress, viral infection, and carcinogenesis. The present study analyzed the level of total antioxidant status (TAS) as well as the activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) in patients with oropharyngeal cancer both Epstein-Barr virus (EBV)-positive and EBV-negative in comparison with the control group. The correlations between these parameters and EBV type (wild-type LMP1 (wt-LMP1) or LMP1 with deletion (del-LMP1)), level of antibodies against EBV, the degree of tumor differentiation, and TNM classification were also investigated. Fresh frozen tumor tissue samples collected from 66 patients with oropharyngeal squamous cell carcinoma were tested using nested PCR assay for EBV DNA detection. Spectrophotometric methods were used to measure TAS values as well as SOD and GPx activities in homogenates of tissue, using diagnostic kits produced by Randox Laboratories. Sera from all individuals were investigated using ELISA method to detect the presence of Epstein-Barr virus capsid antigen (EBVCA) IgM and IgG, Epstein-Barr virus nuclear antigen (EBNA) IgG, and early antigen (EA) IgG antibodies. The level of TAS and activities of antioxidant enzymes (GPx and SOD) were significantly decreased in tissues with oropharyngeal cancer, particularly in EBV-positive cases. In 82.3% of patients, wt-LMP1 was detected. Significantly lower TAS, GPx, and SOD values were stated in patients infected with wild-type EBV. The presence of antibodies against early antigen (anti-EA) was detected in over 80% of patients, which suggests reactivation of EBV infection. The correlation between the degree of tumor differentiation and TN classification, especially in EBV-positive patients, was also observed. Determination of these parameters may be useful in evaluating tumor burden in patients with various stages of oropharyngeal cancer and could be an important prognostic factor. Future studies are needed to understand the role of EBV lytic reactivation induced by oxidative stress.

Serum EBV antibodies and LMP-1 in Polish patients with oropharyngeal and laryngeal cancer.

BACKGROUND: The association between Epstein-Barr virus (EBV) and the development of head and neck cancer was reported by many researchers. The aim of the present study was to detect EBV DNA and EBV antibodies in 110 Polish patients with oropharyngeal and laryngeal cancer compared to 40 healthy individuals. METHODS: Frozen tumor tissue fragments were tested using nested PCR assay for EBV DNA detection. Sera from all individuals were investigated using ELISA tests to detect the presence of VCA IgM and IgG, EBNA IgG, EA IgG. RESULTS: EBV DNA was detected in 52.7% of the patients (25% in controls). EBVCA were detected in 94.5%, EBNA in 96.4% and EA in 94.5% of patients. The significantly higher level of EA in the patients suggests EBV reactivation. The majority of patients (83%) were infected with wild-type EBV. CONCLUSION: Our study showed that this variant seems to be associated with oropharyngeal and laryngeal cancer in the Polish population.

Salivary and serum IL-10, TNF-α, TGF-ß, VEGF levels in oropharyngeal squamous cell carcinoma and correlation with HPV and EBV infections.

BACKGROUND: Each year approximately 6,000 new cases of head and neck cancer are registered in Poland. Human papillomavirus (HPV) and Epstein-Barr virus (EBV) have been associated with tumour formation. Cytokines have been shown to play an important role both in inflammation and carcinogenesis and they can be detected in saliva and serum with ELISA assays. Salivary biomarkers may be used as markers of early cancer detection. The aim of this study was the analysis of the serum and salivary levels of IL-10, TNF-α, TGF-ß and VEGF in patients with oropharyngeal cancer and in healthy individuals. The level of these biomarkers was also analyzed in HPV- and EBV-related cases. METHODS: The study involved 78 patients with histopathologically confirmed oropharyngeal squamous cell carcinoma and 40 healthy controls. Serum and salivary levels of IL-10, TNF-α, TGF-ß and VEGF were analyzed both in patients and in healthy individuals by ELISA method using Diaclone SAS commercially available kits (France). EBV DNA was detected by the nested PCR for amplification of EBNA-2. HPV detection and genotyping was performed using the INNO-LiPA HPV Genotyping Extraassay (Innogenetics N. V, Gent, Belgium). The obtained results were subjected to statistical analysis using Mann-Whitney and Kruskal Wallis tests. Test values of p < 0.05 were considered statistically significant. RESULTS: The level of tested cytokines was higher in patients than in controls both in serum (IL-10: 2.3 pg/ml vs 1.65 pg/ml, p = 0.0003; TGF-ß: 11.3 ng/ml vs 7.8 ng/ml, p = 0.0005; VEGF: 614 pg/ml vs 210 pg/ml, p = 0.0004; TNF-α: 15.0 ng/ml vs 12.90 ng/ml, p = 0.1397) as well as in saliva (IL-10: 5.9 pg/ml vs 2.5 pg/ml, p = 0.00002; TGF-ß: 24.1 ng/ml vs 14.8 ng/ml, p = 0.00002; VEGF: 4321 pg/ml vs 280 pg/ml, p = 0.0000; TNF-α: 23.1 ng/ml vs 11.3 ng/ml, p = 0.00002). EBV DNA was detected in 51.3 % of patients and 20 % of controls, HPV DNA was present in 30.8 % of patients and 2,5 % of controls. The level of IL-10 was statistically higher in patients infected with EBV, HPV and co-infected with EBV/HPV. The level of TNF-α was significantly higher in patients infected with EBV, while TGF-ß in patients with HPV infection and EBV/HPV co-infection. CONCLUSION: Detection of salivary cytokines may be very helpful in early diagnosis, treatment and prognosis of OSCC.