Glutathione (GSH) is the keystone of the cellular response toward oxidative stress. Elevated GSH content correlates with increased resistance to chemotherapy and radiotherapy of head and neck (HN) tumors. The purpose of the present cross-sectional study was to evaluate whether the expression of glutamate-cysteine ligase (GCL) accounts for the increased GSH availability observed in HN squamous cell carcinoma (SCC). For that purpose, the messenger (m)RNA levels of the modifier (M) and catalytic (C) subunits of GCL and its putative regulators (namely, nuclear factor erythroid 2-related factor 2, heme oxygenase-1 and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha) were monitored in 35 surgical resections of untreated HNSCC. The localization of GCLM was evaluated using in situ hybridization and immunohistochemistry. GCLM expression was significantly increased in tumor samples, compared with normal mucosa, both at the mRNA and protein level (P=0.029), but the pathway of GCLM activation remains to be elucidated. Protein expression of GCLM was detected in the cytoplasm and nucleus. GCLM and the proliferation marker Ki-67 displayed a similar distribution, being both mainly expressed at the periphery of tumor lobules. The present study reported increased expression of GCL and the rate-limiting enzyme of GSH synthesis, within HNSCC. The nuclear localization of GCLM and the concomitant expression of Ki-67 suggested that the localization of GSH synthesis contributes to the protection against oxidative stress within hotspots of cell proliferation.

Lessons Learned Cabazitaxel has activity in squamous cell carcinoma of the head and neck (SCCHN) and taxane-resistant cell lines. For the first time, cabazitaxel was investigated in incurable patients with recurrent SCCHN. Patients were randomly assigned to cabazitaxel every 3 weeks or weekly methotrexate. This phase II study did not meet its primary endpoint. Cabazitaxel has low activity in SCCHN. The toxicity profile in this population also was not favorable owing to the high rate of febrile neutropenia observed (17%). Background. Cabazitaxel is a second-generation taxane that improves the survival of patients with metastatic castrate-resistant prostate cancer following docetaxel therapy. Cabazitaxel has activity in squamous cell carcinoma of the head and neck (SCCHN) and taxane-resistant cell lines. In this randomized phase II trial, we investigated cabazitaxel in patients with recurrent SCCHN. Methods. Patients with incurable SCCHN with progression after platinum-based therapy were randomly assigned to cabazitaxel every 3 weeks (cycle 1, 20 mg/m2, increased to 25 mg/m2 for subsequent cycles in the absence of nonhematological adverse events [AEs] greater than grade 2 and hematological AEs greater than grade 3) or methotrexate (40 mg/m2/week). The patients were stratified according to their performance status and previous platinum-based chemotherapy for palliation versus curative intent. The primary endpoint was the progression-free survival rate (PFSR) at 18 weeks. Results. Of the 101 patients, 53 and 48, with a median age of 58.0 years (range, 41–80), were randomly assigned to cabazitaxel or methotrexate, respectively. The PFSR at 18 weeks was 13.2% (95% confidence interval [CI], 5%–25%) for cabazitaxel and 8.3% (95% CI, 2%–20%) for methotrexate. The median progression-free survival was 1.9 months in both arms. The median overall survival was 5.0 and 3.6 months for cabazitaxel and methotrexate, respectively. More patients experienced serious adverse events with cabazitaxel than with methotrexate (54% vs. 36%). The most common drug-related grade 3–4 AE in the cabazitaxel arm was febrile neutropenia (17.3%). Conclusion. This study did not meet its primary endpoint. Cabazitaxel has low activity in recurrent SCCHN.

Over 90% of head and neck cancers are squamous cell carcinomas (HNSCC) and the overall 5-year survival rate is up to 50%. The redox status of these cancers is an important factor in carcinogenesis and plays a role in radioresistance and therefore locoregional recurrences. However, knowledge of the redox status is rather limited. Glutathione is the major reactive oxygen species scavenger in normal cells. We compared the levels of tissue redox potential in HNSCC tumor tissue and compared them with those of the adjacent, histologically cancer-free, mucosa. A total of 36 patients with HNSCC were included in the study. The redox status of tumor and normal adjacent tissue was measured by the oxidized/reduced glutathione (GSSG/GSH) ratio in capillary electrophoresis. The GSSG/GSH ratio in the tumor tissue was lower compared with adjacent normal tissue in 38% of the patients. Pretherapy HNSCC tumor tissue has variable GSH levels compared with adjacent cancer-free mucosa. This difference was not related to clinical and pathological parameters. Further studies are required to determine whether the GSSG/GSH ratio plays a role in carcinogenesis and could predict radioresistance.

Two cases of severe skin reactions to cetuximab and concomitant radiation therapy are reported in patients treated for locally advanced head-and-neck squamous cell carcinoma and concurrently treated with an antivitamin K drug for cardiovascular diseases. The role of vitamin K in the epidermal growth factor/epidermal growth factor receptor pathway is discussed. A plea for a specific registry is entered.

BACKGROUND:Blood fluidity is maintained by a delicate balance between coagulation and fibrinolysis. The endothelial cell surface is a key player in this equilibrium and cell surface disruptions can upset the balance. We investigated the role of pericellular myeloperoxidase oxidized LDLs (Mox-LDLs) in this balance.METHODS AND RESULTS:We designed a technical device that enabled us to monitor fibrinolysis in real-time at the surface of an endothelial cell line (EA.hy926), and showed that Mox-LDL decreased pericellular fibrinolysis. There were no changes in fibrinolysis when EA.hy926 endothelial cells were exposed to native LDL (24 hours) at doses of 10, 50, 100 and up to 1250 µg/ml. However, treatment of EA.hy926 endothelial cells with 10 and 50 µg/ml of Mox-LDL (physiological serum concentrations) increased the lysis time by 15 and 13%, respectively (p<0.001), although this effect was not present at higher concentrations of 100 µg/ml. This effect was not correlated with any changes in PAI-1 or t-PA or PA Receptor (PAR) expression. No effect was observed at the surface of smooth muscle cells used as controls.CONCLUSION:Our data link the current favorite hypothesis that modified LDL has a causal role in atheroma plaque formation with an old suggestion that fibrin may also play a causal role. Our data help complete the paradigm of atherosclerosis: Modified LDL locally enhances fibrin deposition (present work); fibrin deposits enhance endothelial permeability; this effect allows subendothelial accumulation of lipid and foam cells.

Introduction : The epidemiology and clinical picture of laryngeal and hypopharyngeal carcinomas have changed significantly in the past 50 years. The aim of this study was to analyze selected epidemiological and clinical characteristics of laryngeal and hypopharyngeal carcinoma.Methods : A complete chart review of all patients records was conducted. All the patients who were diagnosed as having laryngeal or hypopharyngeal cancer from January 1, 2004 through December 31, 2009 were included in the study. The demographics of the patient population, the disease profile were analyzed.Results : 138 patients with laryngeal or hypopharyngeal disease were treated. 76 patients presented a laryngeal cancer. Disease characteristics indicated that most cases of supraglottic cancer were in a locally advanced stage (84,4%), whereas most patients with glottis cancer were diagnosed with early stage (63,3%). A hypopharyngeal cancer was diagnosed in 62 cases. A significant increasing trend in hypopharyngeal cancer has been seen in males. The majority of the patients was alcohol consumers and had a histology showing squamous cell carcinoma. There were 33 females and 105 males whose ages ranged at presentation from 47 to 86 years.Of the 138 patients treated, 24 and 47 patients were respectively T3 and T4 and 37 patients were N1, 37 patients N2 and 10 patients N3 (Table I). Most patients had stage IV disease (65/138). Majority of cases presented with local advanced stage. Of the 138 patients treated, 24 and 47 patients were respectively T3 and T4. The highest rate of local advanced stage was observed in patients with pyriform sinus carcinomas (81%); the lowest rate was observed for glottis tumors (41,8%). Regional lymph node metastases were diagnosed in 61% of the analyzed cases.37 patients were N1, 37 patients N2 and 10 patients N3. The highest rate (82,2%) of regional lymph node metastases were observed in cases of pyriform sinus carcinomas, and the lowest (31,7%) in glottis carcinomas. Most patients had stage IV disease (65/138).49 patients received radiotherapy; 48 patients were treated by surgery followed by(chemo)radiotherapy. 41 patients were treated initially by concomitant chemoradiotherapy.Conclusion : A tendency for increasingly younger patients to develop larynx and hypopharynx carcinomas was observed. Most patients had stage IV disease but no trend for a percentage increase in locally advanced tumors was observed. A significant increasing trend in hypopharyngeal cancer has been seen in males.

OBJECTIVES:Many studies have evaluated the ways in which sleep disturbances may influence inflammation and the possible links of this effect to cardiovascular risk. Our objective was to investigate the effects of chronic sleep restriction and recovery on several blood cardiovascular biomarkers.METHODS AND RESULTS:Nine healthy male non-smokers, aged 22-29 years, were admitted to the Sleep Laboratory for 11 days and nights under continuous electroencephalogram polysomnography. The study consisted of three baseline nights of 8 hours sleep (from 11 pm to 7 am), five sleep-restricted nights, during which sleep was allowed only between 1 am and 6 am, and three recovery nights of 8 hours sleep (11 pm to 7 am). Myeloperoxidase-modified low-density lipoprotein levels increased during the sleep-restricted period indicating an oxidative stress. A significant increase in the quantity of slow-wave sleep was measured during the first recovery night. After this first recovery night, insulin-like growth factor-1 levels increased and myeloperoxidase concentration peaked.CONCLUSIONS:We observed for the first time that sleep restriction and the recovery process are associated with differential changes in blood biomarkers of cardiovascular disease.

Raloxifene (RLX), a selective oestrogen receptor modulator, has oestrogen-agonist effects on bone, lipoproteins, and homocysteine and oestrogen-antagonist activity in the breast and uterus, positioning it as a potential drug for long-term prevention of coronary heart disease in postmenopausal women. To further evaluate its influence on cardiovascular risk factors, we studied the effects of 60 mg/day RLX on serum lipid levels, inflammatory (high-sensitivity C-reactive protein, and coagulation (fibrinogen) markers, monocytes, and fibrinolysis in 15 healthy postmenopausal women. Markers were measured at baseline, after 1 month without treatment, and after 3 months of treatment. Fibrinolysis was evaluated using the euglobulin clot lysis time (ECLT) determined with a new semiautomatic optical method. Monocyte phenotype was determined by measurement of the expression of the antigens CD14, HLA-DR, and CD62-L using flow cytometry. After 3 months of RLX treatment, we observed a decrease in total cholesterol (p = 0.002), in low-density lipoprotein cholesterol (p <0.001), and in lipoprotein A (p = 0.01). Fibrinogen (p = 0.002) decreased significantly, and high-sensitivity C-reactive protein had a tendency to decrease, but this did not reach statistical significance (p = 0.06). RLX treatment had no effect on ECLT (p = 0.223) or on white blood cell, lymphocyte, and total monocyte counts (p = 0.313). Monocyte expression of HLA-DR, CD14, and CD62-L was not modified by the treatment. In conclusion, we confirm that RLX has beneficial short-term effects on levels of lipids and inflammatory markers, with no effect on fibrinolysis or monocyte phenotype.

Due to imprecise clinical staging, the finding of extravesical and node-positive disease at the time of radical cystectomy (RC) for patients with clinically localized bladder cancer is not uncommon. Circulating tumor cells (CTCs) have been shown to be present in the peripheral blood of patients with metastatic urothelial carcinoma. The object of this study was to evaluate the ability of CTCs to predict extravesical disease in bladder cancer patients prior to RC.Peripheral blood samples from 43 patients with bladder cancer were evaluated using the CellSearch (Veridex, LLC, Raritan, NJ) CTC assay prior to RC. The sensitivity, specificity, and positive predictive value (PPV) of CTC status in predicting extravesical disease was calculated. Receiver operating characteristic (ROC) curves were generated to quantify the ability of CTCs to predict extravesical and node-positive disease.CTCs were detected in 9 (21%) patients prior to RC. The sensitivity, specificity, and PPV of CTC status in predicting extravesical disease were 27%, 88% and 78%, respectively. The accuracy of CTC status in predicting extravesical (≥pT3 or node-positive) disease for the entire cohort was 0.576. In a model incorporating preoperative hydronephrosis, CTC status did not improve the predictive accuracy for extravesical disease (0.576 vs. 0.585, P = 0.915).CTCs were detected in low numbers in a small percentage (21%) of patients prior to undergoing RC at our institution. CTC status was not a robust predictor of extravesical or node-positive disease in this cohort. CTC status is not likely to be a clinically useful parameter for directing therapeutic decisions in patients with ≤cT2 bladder cancer.

This review focuses on the immunohistochemical expression of members of the MUC-type mucin family in salivary gland tumors and head and neck squamous cell carcinomas (HNSCC). Information is available on changes in the expression levels and distribution profiles of MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC6 and MUC7 in tumors of the salivary glands; and of MUC1, MUC2 and MUC4 in HNSCC. In salivary gland tumors the expression patterns of MUC2, MUC3, MUC5AC and MUC6 appear to be very closely correlated with the histopathological tumor type indicating their potential use to improve diagnostic accuracy in salivary gland neoplasia. Some MUC-type mucins have emerged as valuable prognostic indicators in pleomorphic adenoma, mucoepidermoid carcinoma and HNSCC. Nine antibodies directed against different MUC1 antigens have thus far been examined in HNSCC of which monoclonal antibodies DF3, HMFG-1 and Ma695 have shown significant correlations with disease outcome. The importance of taking the specific anti-MUC antibody into consideration when comparing the results of different studies on MUC expression in salivary gland tumors and HNSCC is also highlighted in this review.

Objective: To investigate the influence of neuraminidase, an enzyme that cleaves sialic acid from the red blood cell (RBC) membrane, on RBC shape and biochemistry in critically ill patients. Design: Prospective, observational study and in vitro laboratory study. Setting: A 31-bed medico-surgical department of intensive care and a university-affiliated cell biology laboratory. Subjects: Acutely ill patients with and without sepsis and healthy volunteers. Interventions: Blood sampling in volunteers. Measurements and Main Results: Neuraminidase activity was measured using a fluorescent assay. RBC shape was assessed by the second coefficient of dissymmetry of Pearson using a flow cytometry technique at 25°C. Intraerythrocytic 2,3-diphosphoglycerate and lactate contents were also measured. Neuraminidase activity was significantly higher in septic patients compared with nonseptic patients and healthy volunteers (5.42 [4.85–6.00] vs. 4.53 [4.23–5.23] and 1.26 [0.83–1.83] mU/mL; all p < 0.05). Neuraminidase treatment modified the RBC shape in vitro in a dose–response fashion, and most of these alterations were present after 10 hours of incubation. Incubation of RBCs with phosphatidylinositol phospholipase C modified RBC shape and increased sialic acid concentrations in the supernatant, suggesting a leakage of neuraminidase from the RBC membrane. Alterations in shape were associated with increased 2,3-diphosphoglycerate (0.46 ± 0.25 vs. 0.19 ± 0.05 &mgr;mol/mL; p = 0.006) and lactate content (0.81 ± 0.07 vs. 0.66 ± 0.05 mmoL/L; p = 0.002). Conclusions: In sepsis, desialylation under the influence of increased neuraminidase activity may contribute to the alterations in RBC rheology. Inhibition of neuraminidase may represent a new therapeutic option to ameliorate RBC rheology and perhaps oxygen delivery to the cells.

BACKGROUND:Endothelial cell dysfunction, by promoting fibrin deposition, has been implicated in the development of multiple organ failure. Altered fibrinolysis during inflammation may participate in microvascular alterations. We sought to determine whether plasma fibrinolysis was related to the severity of organ dysfunction and/or to the levels of von Willebrand factor (vWF antigen), as a marker of endothelium dysfunction, in critically ill patients.METHODS:Forty-nine consecutive patients admitted to an adult medico-surgical intensive care unit (ICU) with (18) or without sepsis (31) were included. C-reactive protein and vWF levels were measured on ICU admission and plasma fibrinolysis was assessed by the Euglobulin Clot Lysis Time (ECLT). The sequential organ failure assessment (SOFA) score and the simplified acute physiology score (SAPS) II were calculated on admission.RESULTS:ECLT was significantly longer in septic than in non-septic patients [1033 min (871-1372) versus 665 min (551-862), p = 0.001]. There were significant correlations between ECLT and C-reactive protein (CRP) concentrations (r = 0.78, p < 0.001) and the Sequential Organ Failure Assessment (SOFA) score (r = 0.39, p = 0.006). The level of vWF was not correlated with the ECLT (r = -0.06, p = 0.65) or the SOFA score (r = -0.02, p = 0.88).CONCLUSION:ECLT measurement at admission could be a marker of organ dysfunction and a prognostic indicator in critically ill patients.

L-asparaginase is commonly used in the chemotherapy regimens for acute lymphoblastic leukaemia. Its use is associated with thrombotic complications in 1 to 14 % of the cases. The pathogenesis of this complication is still unclear. However, the decrease of antithrombin seems to play an important role. We report a case of a 17-year old man with a acute lymphoblastic leukaemia, who developed a cerebral sinovenous thrombosis due to an acquired deficiency of antithrombin and protein C and S following L-asparaginase chemotherapy. We discuss the use of prophylactic supplements of antithrombin and the value of screening of thrombophilia based on the recent medical literature.

OBJECTIVES:This study examines the effects of sleep restricted to four hours for three consecutive nights on blood parameters, known to be associated with cardiovascular risk, in young healthy men. MATERIAL AND METHODS:Eight young healthy men (age 24.5 +/- 3.3 years) were studied in the sleep restricted group. Nine young healthy men (age 24 +/- 2 years) were included in the control group and spent the days and nights in the sleep lab, while sleeping eight hours/night. One baseline night was followed by three nights of sleep restriction to four hours and by one recovery night of eight hours. Blood samplings were performed after the baseline night and after the third night of sleep restriction or without restriction for the control group. RESULTS:A significant increase in white blood cells (WBC) (5.79 +/- 1.05 vs. 6.89 +/- 1.31 10(3) cell/microl, p = 0.03), and neutrophils (3.17 +/- 0.69 vs 4.24 +/- 0.97 10(3) cell/microl, p = 0.01) was observed after the third night of sleep restriction. Other blood parameters were not affected. No significant variation was observed in the control group. CONCLUSION:Sleep restriction affected WBC count, mainly neutrophils, considered as risk factor for cardiovascular disease. Stress induced by the short term sleep restriction could be involved in this observation.