The TSPO Ligands MGV-1 and 2-Cl-MGV-1 Differentially Inhibit the Cigarette Smoke-Induced Cytotoxicity to H1299 Lung Cancer Cells.

TSPO is involved in cigarette smoke (CS)-induced cellular toxicity, which may result in oral and pulmonary diseases and lung cancer. H1299 lung cancer cells were exposed directly to CS. The H1299 cells were pretreated with our TSPO ligands MGV-1 and 2-Cl-MGV-1 (Ki = 825 nM for both) at a concentration of 25 µM 24 h prior to CS exposure. Cell death and apoptotic markers were measured, in addition to TSPO expression levels, ATP synthase activity, generation of reactive oxygen species (ROS), depolarization of mitochondrial membrane potential (ΔΨm), cAMP and LDH levels. Pretreatment with MGV-1 and 2-Cl-MGV-1 (25 µM), 24 h prior to CS exposure, differentially attenuated the CS-induced cellular insult as well as cell death in H1299 lung cancer cells. These protective effects included prevention of ATP synthase reversal, ROS generation, depolarization of the mitochondrial membrane and elevation in LDH. The preventive efficacy of 2-Cl-MGV-1 was superior to that achieved by MGV-1. Both ligands did not prevent the elevation in cAMP. These findings may indicate a mild protective effect of these TSPO ligands in CS-related pulmonary and keratinocyte cellular pathology.

Transarterial Embolization in Maxillofacial Intractable Potentially Life-Threatening Hemorrhage.

PURPOSE: Although transarterial embolization (TAE) of vascular lesions with embolizing agents through angiographic catheters has been used for more than 45 years, reports of life-threatening maxillofacial bleeding are relatively rare and have not been updated. The authors review treatment modalities, present their experience of the past 21 years, and suggest a comprehensive algorithm and guidelines for the use of TAE in the treatment of intractable life-threatening maxillofacial hemorrhage. MATERIALS AND METHODS: This article describes 28 patients treated with TAE for severe bleeding that did not respond to conservative therapies. Of these, 13 had uncontrolled epistaxis, 9 were oncologic patients, 4 were postsurgical patients, and 2 were trauma patients. RESULTS: Details of patients' medical history, failed conservative therapy administered before TAE, imaging results, and blood vessels involved are presented, as are the TAE procedures and materials used, outcome, and complications. All these are discussed in relation to the available updated literature. All 9 oncologic patients (100%) had been treated with chemotherapy before the uncontrolled bleeding, and 7 also had radiotherapy administered to the maxillofacial region. Continuous anticoagulant therapy also seemed to predict such bleeding episodes. TAE resolved the bleeding in all 28 cases and rapidly in 90% of cases. Only in 3 oncologic cases did continued bleeding require 3 to 4 consecutive TAE sessions and combinations of embolizing agents. CONCLUSIONS: The reported high rate of success could be the result of careful techniques, appropriate preoperative imaging, highly professional personnel, and intraoperative and perioperative treatments.

Carbonyl levels and survival rates in oral cancer cells exposed to cigarette smoke.

BACKGROUND: Cigarette smoke (CS) is the main inducer of oral cancer, increasing prevalence 4-7 times. MATERIALS AND METHODS: We examined SCC-25 and SCC-15 suitability for studying CS effects on oral cancer cells, measuring carbonyl levels for free radical-mediated CS effect on survival and time/CS dependence. RESULTS: Protein oxidation increased significantly during CS exposure. At all time points, carbonyl levels increased six-fold (p<0.001) in both cell lines. Cell viability decrease was time-dependent. Longer CS exposure led to higher cell mortality. At 120 min, SCC-25 cell survival reduction was 43.7%, (p<0.01). Propidium iodide (PI) assay results matched the Trypan blue assay showing a time-dependent cell viability decrease following CS exposure. At 120 min, cell survival reduction was 37% (p<0.05). CONCLUSION: Cell death is mediated by CS free radicals with pathological process occurring first. Oral cancer cell models SCC-25 and SCC-15 are suitable for studying CS-induced free radical-related damage, potentially leading to the pathogenesis of oral cancer.

Cigarette smoke-induced effects on the cell cycle in oral cancer cells: reduction of G2/M fraction.

BACKGROUND: Cigarette smoke (CS) is the main inducer of oral cancer, increasing its prevalence by 4-7 times. MATERIALS AND METHODS: We examined the suitability of cell models SCC-25 and SCC-15 for studying effects of CS on oral cancer cells and whether CS significantly affects the cell cycle using fluorescence-activated cell sorting assays. RESULTS: There was significant change in the fraction of SCC-15 cells in pre-G1 state following CS exposure. At 60 and 90 min, increase in the pre-G1 cell fraction was 118% (p<0.05) and 135% (p<0.01) respectively. The G2/M cell fraction was significantly lower following CS exposure. At 90 and 120 min following CS exposure, the G2/M fraction levels had declined by 44% (p<0.05) and 34% (p<0.01) respectively. Results for SCC-25 cells were similar. At 90 and 120 min following CS exposure, the pre-G1 fraction of the cells increased by 230% and 550%, respectively (p<0.01). At 120 min of CS exposure, the fraction of G2/M cells was lower by 47% (p<0.05) compared to controls. CONCLUSION: CS profoundly affects the cell-cycle distribution in both SCC-15 and SCC-25 oral cancer cellular models. Such effects have been associated with DNA damage and carcinogenesis. Both models are useful for studying oral cancer pathogenesis.

DNA fragmentation induced by cigarette smoke in oral cancer cells.

BACKGROUND/AIM: Cigarette smoke (CS) is the main inducer of oral cancer, increasing the prevalence by 4-7 times. We examined induction of apoptosis by CS exposure of SCC-25 and SCC-15 oral cancer cells. MATERIALS AND METHODS: After controlled exposure to CS of various durations and at different time points we measured DNA fragmentation to assay apoptotic levels. RESULTS: SCC-15 cells showed a 70% (p<0.05) increase in apoptotic levels immediately after 30 min of exposure to CS. Twenty-four hours after 30-min CS exposure a further increase in apoptotic levels to 178% (p<0.05) could be observed. However, SCC-15 cells showed a decrease in apoptotic levels immediately after 180-min exposure to CS. CS-exposed SCC-25 cells did not show such CS-related effects. CONCLUSION: SCC-15 and SCC-25 oral cancer cells respond differently to CS regarding apoptotic cell death levels. In this respect, SCC-15 cells are sensitive to CS, while SCC-25 cells are not. Further comparisons between these cells may give insight regarding relationships between CS, apoptosis and invasiveness of oral cancer.

Salivary and serum analysis in children diagnosed with pneumonia.

The aim of the current study was to evaluate specific markers for pneumonia by using a non-invasive assessment of inflammatory/oxidative biomarkers in saliva accompanying a routine serum analysis. No study evaluating saliva of children with pneumonia has been published previously. Salivary analysis was performed in 15 children diagnosed with lobar pneumonia and in a parallel group of 16 children matching in age and gender in whom there was no respiratory illness, and compared to the serum analysis obtained routinely in both groups of children. Salivary flow rate was lower in the patients' group as was uric acid concentration (by 60%). Increase in salivary concentrations of almost all parameters analyzed was found: Ca, P, and Mg concentrations were higher in the patients' group by 23%, 55%, and 33%, respectively, while LDH, total protein amylase and albumin concentrations were higher by 275%, 79%, and 42%, respectively. In the serum, white cell counts and neutrophils were significantly higher, and sodium level significantly lower in the patients' group. Compositional changes were in the range of 3-80% while the saliva alterations were more profound, in the range of 42-275%. The results demonstrated in the current study indicate salivary analysis as a potentially novel tool for children with pneumonia. Human salivary collection and analysis is a non-invasive tool that could provide additional information for diagnosis and follow-up of pneumonia, especially in children. This is especially beneficial for pediatric patients, as salivary collection is simple, non-invasive, and patient-friendly.

The heart team approach to coronary revascularization--have we crossed the lines of evidence-based medicine?

Evidence-based medicine demands considerable time and decision-making skills to navigate through the proliferating data. A hierarchical "pyramid of evidence" has been formulated to help categorize data quality. The hierarchical data are processed into recommendations in Practice Guideline statements. Recently, both American College of Cardiology/American Heart Association/Society for Cardiac Angiography and Interventions and European Society of Cardiology guidelines for percutaneous coronary intervention embraced a new "heart team approach" as the preferred method to optimize revascularization decision making in cases of complex coronary anatomy. This extrapolation of a research method to the broad clinical practice has potential limitations. We suggest that both the need for a new method to optimize patient triage for the various revascularization strategies and the method to optimize decision making should be discussed. Published data suggest only minor deviations from guideline-based indications. Furthermore, traditional clinical judgment may result in a better patient outcome than arbitrary treatment assignment by rigid set of criteria. In conclusion, the need for a new decision-making process in the choice of revascularization strategy should be further explored and supported by scientific evidence.

Salivary biomarker analysis complementing regular clinical examination.

AIM: Tools for the diagnosis and evaluation of the severity and prognosis of periodontitis are currently insufficient. The aim of the current study was to find an array of salivary biomarkers that would be both sensitive and specific enough to be used as a complement to regular clinical examination. Furthermore, we investigated salivary markers of successful periodontal treatment to elucidate the underlying mechanism of disease. PATIENTS & METHODS: Saliva was collected from periodontitis patients during illness and following successful disease termination. Parameters measured included sodium, potassium, chloride, calcium, phosphate, magnesium, total protein, Alb, LDH, amylase, IL-1α, uric acid and carbonyls. RESULTS: Seven of the parameters analyzed were significantly altered when the disease is active and may be considered biomarkers. CONCLUSION: Salivary composition reflects disease severity and response to therapy, suggesting the efficacy of saliva monitoring for periodontal disease status and care.

Is human saliva an indicator of the adverse health effects of using mobile phones?

Increasing use of mobile phones creates growing concerns regarding harmful effects of radiofrequency nonionizing electromagnetic radiation on human tissues located close to the ear, where phones are commonly held for long periods of time. We studied 20 subjects in the mobile-phone group who had a mean duration of mobile phone use of 12.5 years (range 8-15) and a mean time use of 29.6 h per month (range 8-100). Deaf individuals served as controls. We compared salivary outcomes (secretion, oxidative damage indices, flow rate, and composition) between mobile phone users and nonusers. We report a significant increase in all salivary oxidative stress indices studied in mobile phone users. Salivary flow, total protein, albumin, and amylase activity were decreased in mobile phone users. These observations lead to the hypothesis that the use of mobile phones may cause oxidative stress and modify salivary function.

The prognostic role of phospho-Src family kinase analysis in tongue cancer.

BACKGROUND: The up regulation of Phospho-Src family kinase oncogene has been correlated with reduced postoperative survival in various cancers but never in tongue cancer. METHODS: We analyzed phospho-Src family kinase in 39 tongue (mobile) cancer patients by immunohistochemistry, compared these results with similar analysis for TUNEL and c-erbB-2 and with both clinical tumor characteristics and patient survival probability rates. RESULTS: Phospho-Src family kinase overexpression was found in most tongue cancer biopsies (62%), significantly correlating with tumors larger in size (P = 0.05), progression­lymph node metastasis (0.004) and stage (P = 0.05), and correlating with TUNEL (P = 0.01) and c-erbB-2 (P = 0.05) expression rates. At 60 months, survival probability for negative phospho-Src family kinase level (=0) patients was 67%, but 30% for positive phospho-Src family kinase level (>0) patients (P = 0.05). CONCLUSIONS: Inverse correlation between phospho-Src family kinase and patient survival demonstrates the prognostic role of phospho-Src family kinase in tongue cancer. These findings suggest a novel link between phospho-Src family kinase and TUNEL and c-erbB-2 pathways, tilting the balance toward cell proliferation.

Saliva as a tool for oral cancer diagnosis and prognosis.

Saliva testing, a non-invasive alternative to serum testing, may be an effective modality for diagnosis and for prognosis prediction of oral cancer, as well as for monitoring post therapy status, by measuring specific salivary macromolecules, examining proteomic or genomic targets such as enzymes, cytokines, growth factors, metalloproteinases, endothelin, telomerase, cytokeratines, mRNA's and DNA transcripts. Salivary analysis has been shown to be a useful diagnostic tool also for distant malignancies such as breast cancer. In recent years, significant alterations have been demonstrated in the saliva of oral cancer patients in the epithelial tumor markers--Cyfra 21-1, TPS and CA12, various oxidative stress-related salivary parameters as ROS and RNS, biochemical and immunological parameters as IGF and MMP's and RNA transcripts of IL8, IL-1B, DUSP1, HA3, OAZ1, S100P, and SAT. Collectively these accumulated data are predicted to alter the field of oral cancer diagnosis by employing highly sensitive new tools which will enable both medical professionals and the patients themselves to monitor their saliva for diagnosis and prognosis prediction, as they relate to oral cancer. At this point however, the aim of salivary analysis is mainly for screening which may be helpful in the future.

Foreign body granulomas after injection of Bio-alcamid for lip augmentation.

Bio-alcamid is one of the newest agents on the market for soft tissue augmentation. Seven studies were documented in the medical literature that examined the safety of Bio-alcamid (Polymekon, Brindisy, Italy); all reported no cases of tissue migration, foreign body granulomas, allergenicity, or interference with the control of cell proliferation. On 2 separate occasions, a woman who had recently undergone lip augmentation presented at our hospital with submucosal nodules of the lip. Histologic examination revealed multiple foreign body-type granulomas composed of giant cells, epithelioid cells, and chronic inflammation of the lip. Efforts to produce a cosmetic material that fulfills all the criteria as an "ideal" agent has not yet been found because all injectable foreign agents have the potential to induce adverse reactions. Caution must be exercised in all cases and the risks explained to the patient before its use.

The expression and prognostic significance of Cks1 in salivary cancer.

Cks1 is an essential factor in facilitating Skp2-dependent degradation of p27, but its role in salivary malignancies is unknown. Expression of cyclin-dependent kinase subunit 1 (Cks1) was examined in 64 salivary malignancies, compared with p27, S-phase kinase protein 2 (Skp2), Ki-67, p53, and TDT-mediated dutp-biotin nick end labeling (TUNEL) expression, and with THE patient's clinical and pathological parameters. Cks1 expression was markedly increased in 30 patients (47%) and strongly correlated with increased expression of Skp2, Ki-67, p53, and TUNEL, but inversely with p27 levels. High expression of Cks1 WAS strongly associated with lymph node metastases and poor prognosis and survival. Cks1 alterations may have a significant biological role in the pathogenesis of salivary cancer.

The salivary tip of the p53 mutagenesis iceberg: novel insights.

Salivary malignancies are rare, heterogeneous, unpredictable in their clinical behavior and seldom studied. This study focused on examining the expression of mutated p53, the most prevalent mutated gene related to human cancer, in a rather large cohort of salivary malignancies (n = 70) and for a prolonged period (20 years). P53 was found to be a most powerful predictor for poor survival and more so when the tumor concurrently expressed TUNEL and heparanase markers, dramatically dropping the survival probability of the patients to 0! Survival probability at 6 years for patients with tumors stained negatively vs. positively for p53, TUNEL and heparanase was 100% vs. 49% while at 18 years this probability dropped to 67% vs. 0%, respectively (p = 0.023). Significant correlation rates were found between age and poor survival, age and p53, and p53 and other co-existing malignancies. These findings support mutated p53 as a prognostic predictor and a pivotal player in salivary carcinogenesis. Significantly more extensive therapy applied to salivary p53-positive patients did not improve mortality rate, questioning the justification for such extensive therapy and emphasizing the need to understand p53, TUNEL and heparanase biological pathways and develop additional therapeutic tools for fighting salivary cancer.

p27 and salivary cancer.

This study examined p27 expression in a cohort of salivary malignancies (n = 74) for a prolonged period (20 years). Reduction of p27 expression was found to be a most powerful predictor for poor survival and more so when the tumor concurrently expressed high levels of p53, TUNEL and heparanase markers, dramatically dropping the patient survival probability to 0! While no patient whose tumor-staining profile included: p27 > 50%, p53 = 0, TUNEL = 0 and heparanase = 0, died of the disease during the 20-year follow up, the median of survival of the group with p27 0, TUNEL > 0 and heparanase > 0 was only 39 months. The survival probabilities of these two groups at 5 years were 100 and 50%, respectively, and at 20 years they were 100 and 0%, respectively (P = 0.05). Significant p27 reduction also resulted in significantly larger tumor size (T value), higher spread of neck metastasis and extra capsular spread and in more advanced disease (higher stage). Significant correlation rates were found between age and poor survival, age and reduced p27 expression, and reduced p27 expression and other general co-existing malignancies, indicating p27 reduction as part of a general phenomenon-age related mutagenesis. Significantly more extensive therapy applied to patients with salivary reduced-p27 tumors could not prevent the rise in mortality rate, questioning the justification for extensive therapy which is naturally accompanied by higher morbidity. Additional therapeutic tools for fighting salivary cancer, possibly based on the new understanding of the p27, p53, TUNEL and heparanase carcinogenic network, are necessary.

Skp2 and salivary cancer.

Salivary malignancies are rare, heterogeneous, unpredictable in clinical behavior and seldom studied. Skp2 expression was examined in salivary malignancies (n = 75) for a prolonged period (20 years). In 40/75 (53%) cases Skp2 expression rate (staining level) was < or =4% while in the remainder (47%) it was >4%. Correlation between enhanced Skp2 and enhanced p53 staining levels was significant (p = 0.042), as were correlation rates between enhanced Skp2 and reduced p27 staining levels (p = 0.01) and enhanced Skp2 and enhanced TUNEL staining levels (p = 0.008). Survival probability rates dropped when Skp2 expression increased. Median patient survival for reduced-stained-tumor patients (< or =4%) was 143 months and significantly lower, 49 months (p = 0.016), for enhanced-stained-tumor patients (>4%). Survival probability at 5 years was 82% for the former group (< or =4%) and 47% for the latter (>4%). At 20 years, survival dropped to 35% and 18% respectively (p = 0.016). More extensive and aggressive therapy did not reduce mortality in patients with enhanced Skp2-expressing tumors. Significant correlations between poor survival and significantly altered expression rates of Skp2, p27, p53, TUNEL and heparanase in salivary malignancies, suggest a biological role in salivary cancer pathogenesis for these 5 markers. The findings may be used for prognostic and follow-up purposes.

Ki67 and salivary cancer.

We examined Ki67 expression in salivary malignancies of 75 patients with a follow-up period of up to 20 years. Correlations between enhanced Ki67 and enhanced p53 and TUNEL and heparanase staining levels were significant. Median survival for reduced-stained-tumor patients (< or = 5%) was 163 months, dropping significantly to 39 months (p = 0.0005) for enhanced stained tumors (> 5%); 5 year survival probability was 93% and 33%, respectively, 45% and 16%, respectively, (p = 0.0005) at 20 years. Significant correlation between poor survival and concurrently altered expression rates of Ki67 and p53, p27 Skp2, TUNEL and heparanase in the salivary malignancies indicates a biological role in salivary cancer pathogenesis.

TUNEL as a tumor marker of tongue cancer.

Squamous cell carcinoma (SCC) of the tongue has an increasing incidence, a high morbidity rate and a 50% 5-year mortality rate. We analyzed the rate of apoptosis using TDT-mediated dUTP-biotin nick end-labeling (TUNEL), p53 and heparanase in 73 patients with tongue cancer by immonohistochemistry, and tested data for correlation with survival, tumor size, grade and metastasis. TUNEL staining was strong in 54% of the tumors; the remainder lacked staining, as did all healthy control tissues. Significant correlations were found between TUNEL staining level and p53 expression rates (p=0.016) and between TUNEL and heparanase (p=0.039). Moreover, while the cumulative 5-year survival probability for tumors not stained for TUNEL and p53 was 63%, but dropped to 34% with TUNEL staining; for lack of TUNEL and heparanase (=0), 5-year survival rate was 50%, while staining presence (>0) reduced survival to 34%. TUNEL joins other biomarkers in indicating prognosis of understanding pathogenesis of tongue cancer.