Expression profiles of miR-29c, miR-200b and miR-375 in tumour and tumour-adjacent tissues of head and neck cancers.

Altered expression of microRNAs (miRNAs) has been shown in many types of malignancies including the head and neck squamous cell carcinoma (HNSCC). Although there are many new and innovative approaches in the treatment of HNSCC, a clear marker of this disease is still missing. Three candidate miRNAs (miR-29c-3p, miR-200b-5p and miR-375-3p) were studied in connection with HNSCC using quantitative real-time PCR expression levels in 42 tissue samples of HNSCC patients and histologically normal tumour-adjacent tissue samples of these patients. Primary HNSCC carcinoma tissues can be distinguished from histologically normal-matched noncancerous tumour-adjacent tissues based on hsa-miR-375-3p expression (sensitivity 87.5 %, specificity 65 %). Additionally, a significant decrease of hsa-miR-200b-5p expression was revealed in tumour-adjacent tissue samples of patients with node positivity. Lower expression of hsa-miR-200b-5p and hsa-miR-29c-3p in HNSCC tumour tissue was associated with higher tumour grade. Consequently, survival analysis was performed. Lower expression of hsa-miR-29c-3p in tumour-adjacent tissue was associated with worse overall and disease-specific survivals. Lower expression of miR-29c-3p in tumourous tissue was associated with worse relapse-free survival. hsa-miR-375-3p seems to be a relatively promising diagnostic marker in HNSCC but is not suitable for prognosis of patients. Furthermore, this study highlighted the importance of histologically normal tumour-adjacent tissue in HNSCC progress (significant decrease of hsa-miR-200b-5p expression in tumour-adjacent tissue of patients with node positivity and low expression of hsa-miR-29c-3p in HNSCC tumour-adjacent tissue associated with worse prognosis).

Effect of HPV on tumor expression levels of the most commonly used markers in HNSCC.

Approximately 90 % of head and neck cancers are squamous cell carcinomas (HNSCC), and the overall 5-year survival rate is not higher than 50 %. There is much evidence that human papillomavirus (HPV) infection may influence the expression of commonly studied HNSCC markers. Our study was focused on the possible HPV-specificity of molecular markers that could be key players in important steps of cancerogenesis (MKI67, EGF, EGFR, BCL-2, BAX, FOS, JUN, TP53, MT1A, MT2A, VEGFA, FLT1, MMP2, MMP9, and POU5F). qRT-PCR analysis of these selected genes was performed on 74 biopsy samples of tumors from patients with histologically verified HNSCC (22 HPV-, 52 HPV+). Kaplan-Meier analysis was done to determine the relevance of these selected markers for HNSCC prognosis. In conclusion, our study confirms the impact of HPV infection on commonly studied HNSCC markers MT2A, MMP9, FLT1, VEGFA, and POU5F that were more highly expressed in HPV-negative HNSCC patients and also shows the relevance of studied markers in HPV-positive and HPV-negative HNSCC patients.

Prognostic significance of the tumour-adjacent tissue in head and neck cancers.

Even with significant advances in operative skills and adjuvant therapies, the overall survival of patients suffering with head and neck squamous cancers (HNSCC) is unsatisfactory. Accordingly, no clinically useful prognostic biomarkers have been found yet for HNSCC. Many studies analysed the expression of potential markers in tumour tissues compared to adjacent tissues. Nevertheless, due to the sharing of the same microenvironment, adjacent tissues show molecular similarity to tumour tissues. Thus, gene expression patterns of 94 HNSCC tumorous tissues were compared with 31 adjacent tissues and with 10 tonsillectomy specimens of non-cancer individuals. The genes analysed at RNA level using quantitative RT-PCR and correlated with clinico-pathological conditions were as follows: EGF, EGFR, MKI67, BCL2, BAX, FOS, JUN, TP53, VEGF, FLT1, MMP2, MMP9, MT1A and MT2A. The elevated MT2A, BAX, EGF and JUN expression was associated with the influence of tumour cells on the rearrangement of healthy tissues, as well as a significant shift in the BAX/BCL2 ratio. Our investigation also indicated that adjacent tissues play an important role in cancerogenesis by releasing several tumour-supporting factors such as EGF. A gradual increase in the metallothionein expression, from the lowest one in tonsillectomy samples to the highest ones in tumour samples, suggests that MT expression might be tissue reaction to the presence of tumour cells. The results of this study confirmed the significance of metallothionein in tumori-genesis and gave evidences for its use as a potential HNSCC biomarker. Furthermore, this study highlighted the importance of histologically normal tumour-adjacent tissue in prediction of HNSCC progress.

[Importance of taste in maintaining homeostasis and pathological impact of orosensory reflexes distraction in relation to sweet taste after non-caloric sweeteners consumption]. / Význam chuti v udrzení homeostázy organizmu a patologický dopad vyrusení orocefalických reflexu ve vazbe na sladkou chut v následku konzumace nekalorických sladidel.

Taste signals and their reflexes have important signalling function in nature. They protect organism against toxic substances in food with help of taste aversion, they help to cope nutrition deficiencies through taste preferences, on the other hand, they act in many postprandial reflexes to maintain energy homeostasis. It is well-known that sweet taste is important oro-sensory stimulus for mammals. It acts as predictor of caloric food intake even before its entry into stomach and circulation. Taste and other oro-sensory signals from oral cavity affect not only the intake regulation, but also influence hormonal, neural and metabolic pathways to maintain homeostasis. The aim is to utilize effectively food energy and prevent energy instability of organism. Oro-sensory reflexes mediated by taste cells develop naturally from the first contact with sweet breast milk in infancy. It has been proven that the attenuation of reflexes due to the use of artificial sweeteners that don´t bring any caloric value to human body leads to hormonal and energetic dysregulation of organism and may contribute to metabolic syndrome.

High perioperative level of oxidative stress as a prognostic tool for identifying patients with a high risk of recurrence of head and neck squamous cell carcinoma.

BACKGROUND: The aim of the study was to investigate the relationships among (a) glutathione peroxidase (GPx) and malondialdehyde (MDA); (b) oncological characteristics (i.e., TNM classification, tumor grade), and; (c) prognosis of head and neck squamous cell carcinoma. METHODS: In a prospective cohort study, we followed 88 patients for 67.4 months (median 40.3) after surgery for head and neck squamous cell carcinoma. Activity of GPx was determined by ELISA and plasma MDA concentration by liquid chromatography. RESULTS: Lower GPx activity was observed in the T3/4 patients than in the T1/2 group. Tumor grade was significantly correlated with both GPx (P = 0.001) and MDA (P = 0.05, both Spearman). The perioperative level of MDA was higher in patients who later recurred during the follow-up period (n = 15) than in the complete remission group (P = 0.01, Mann-Whitney). Median disease-free interval and overall survival in the group with MDA > median were 29.5 and 32.0 months, respectively, and 38.4 and 40.3 months in the patient group with MDA ≤ median (P = 0.10 and P = 0.08, respectively; Kaplan-Meier). Patients with MDA levels higher than the median had a more than twofold greater risk of recurrence than patients with MDA levels smaller than the median (31.3 vs. 15.2%, P = 0.06, logrank). CONCLUSION: Our results suggest that an increased MDA level at the time of initial surgery is found in patients with a high risk of recurrence, which suggests that each patient can be categorized according to risk of recurrence based on their MDA level at the time of initial surgery.

One-step detection of human papilloma viral infection using quantum dot-nucleotide interaction specificity.

Due to the close relationship between carcinogenesis and human papillomavirus (HPV), and since they are transmitted via huge number of asymptomatic carriers, the detection of HPV is really needed to reduce the risk of developing cancer. According to the best of our knowledge, our study provides the very first method for one-step detection of viral infection and if it has initiated the subsequent cancer proliferation. The proposed novel nanosystem consists of magnetic glass particles (MGPs), which were attached with DNA probe on their surface to hybridize with target DNAs. The MGP-probe-DNA hybrid was finally conjugated with CdTe/ZnSe core/shell quantum dots (QDs). The proposed detection system is based on a novel mechanism in which the MGPs separate out the target DNAs from different biological samples using external magnetic field for better and clear detection and the QDs give different fluorescent maxima for different target DNAs due to their ability to interact differently with different nucleotides. Firstly, the method was optimized using HPV genes cloned into synthetic plasmids. Then it was applied directly on the samples from normal and cancerous cells. After that, the real hospital samples of head and neck squamous cell carcinoma (HNSCC) with or without the infection of HPV were also analyzed. Our novel nano-system is proved successful in detecting and distinguishing between the patients suffering by HPV infection with or without subsequent cancer having detection limit estimated as 1.0 x 109 (GEq/mL). The proposed methodology is faster and cost-effective, which can be applied at the clinical level to help the doctors to decide the strategy of medication that may save the life of the patients with an early treatment.

Prognostic Significance of Serum Free Amino Acids in Head and Neck Cancers.

Despite distinctive advances in the field of head and neck squamous cell cancer (HNSCC) biomarker discovery, the spectrum of clinically useful prognostic serum biomarkers is limited. As metabolic activities in highly proliferative transformed cells are fundamentally different from those in non-transformed cells, specific shifts in concentration of different metabolites may serve as diagnostic or prognostic markers. Blood amino acids have been identified as promising biomarkers in different cancers before, but little is known about this field in HNSCC. Blood amino acid profiles of 140 HNSCC patients were examined using high-performance liquid chromatography. Cox proportional hazards regression model was used to assess the prognostic value of amino acid concentrations in serum. Colony forming assay was used to identify the effect of amino acids that were significant in Cox proportional hazards regression models on colony forming ability of FaDu and Detroit 562 cell lines. In the multivariable Cox regression model for overall survival (OS), palliative treatment was associated with an unfavourable prognosis while high serum levels of methionine have had a positive prognostic impact. In the relapse-free survival (RFS) multivariable model, methionine was similarly identified as a positive prognostic factor, along with tumor localization in the oropharynx. Oral cavity localization and primary radio(chemo)therapy treatment strategy have been linked to poorer RFS. 1mM serine was shown to support the forming of colonies in both tested HNSCC cell lines. Effect of methionine was exactly the opposite.

Oral garenoxacin in the treatment of acute bacterial maxillary sinusitis: a Phase II, multicenter, noncomparative, open-label study in adult patients undergoing sinus aspiration.

BACKGROUND: Garenoxacin is a des-F(6)-quinolone with in vitro activity against key respiratory pathogens, including Streptococcus pneumoniae, Hemophilus influenzae, Staphylococcus aureus, and Moraxella catarrhalis. Limited data are available regarding the effect of garenoxacin in the treatment of acute bacterial sinusitis. OBJECTIVE: The aim of this study was to assess the efficacy and tolerability of garenoxacin in adults with acute bacterial maxillary sinusitis undergoing a pre-treatment diagnostic sinus aspirate. METHODS: This Phase II, multicenter, noncomparative, open-label study was conducted at 30 centers in the United States, Mexico, Argentina, and Europe. Male and female patients aged 18 to 80 years with clinical signs and symptoms lasting >or=5 but or=5 mm because it was believed that improvement in mucosal thickening might not be reliably measurable at the 5-day time point. All patients received garenoxacin 400 mg QD for 5 or 10 days. Maxillary sinus needle aspiration for Gram stain, routine culture, and susceptibility testing were performed before treatment, and, if clinically indicated, during and after treatment. Bacteriologic eradication (negative culture on repeat sinus aspiration) and cure rates (complete resolution of all signs and symptoms) were assessed at a test-of-cure visit 5 to 18 days after the end of treatment. The occurrence of adverse events was recorded by the investigators up to 30 days after the last administration of garenoxacin by questioning patients. RESULTS: A total of 546 patients were enrolled and 543 were randomized (5-day cohort: mean age, 40 years; mean weight, 76 kg; 56% women; 10-day cohort: mean age, 41 years; mean weight, 77 kg; 58% women). Clinically evaluable patients included 253 in the 5-day cohort and 266 in the 10-day cohort. Cure rates were 93% (236/253; 95% CI, 89%-96%) and 91% (243/266; 95% CI, 87%-94%) for evaluable patients in the 5- and 10-day cohorts, respectively. Bacteriologic eradication rates in microbiologically evaluable patients were 94% in both cohorts (5 days, 204/217; 10 days, 182/193). Eradication rates in the 5- and 10-day cohorts were as follows: S pneumoniae, 94% (62/66) and 93% (39/42); H influenzae, 100% (30/30) and 93% (26/28); S aureus, 96% (23/24) and 91% (31/34); and M catarrhalis, 89% (8/9) and 86% (12/14). Of the 9 patients with acute bacterial sinusitis due to multidrug-resistant S pneumoniae, 8 achieved clinical cure with garenoxacin treatment. Adverse events (AEs) most frequently reported were diarrhea (

Establishment of oral squamous cell carcinoma cell line and magnetic bead-based isolation and characterization of its CD90/CD44 subpopulations.

In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44+/CD90-, CD44-/CD90-, CD44+/CD90+, CD44-/CD90-). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90+) and epithelial (CD90-) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44+ cells exhibited stronger migratory capacity than CD44- subpopulations. CD44+ subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44- subpopulations. Furthermore, co-cultivation experiments revealed that CD44-/CD90+ cells supported growth of epithelial tumor cells (CD44+/CD90-). On the contrary, factors released by CD44+/CD90+ type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44-/CD90+, but this subpopulation had a low migratory capacity.

Our experience with benign paroxysmal positional vertigo.

Benign paroxysmal positional vertigo has been considered a separate nosological entity. This status is explained by the theories of cupulolithiasis and canalolithiasis. The disorder is treated with training; success was achieved in 88% of our cases, which correlates with the literary data. Trauma and inflammation in the head and neck region may be regarded as possible etiological factors.

Evaluation of EGFR as a prognostic and diagnostic marker for head and neck squamous cell carcinoma patients.

Approximately 90% of all head and neck tumors are squamous cell carcinomas. The overall survival of patients with head and neck squamous cell carcinoma (HNSCC) is low (≤50%). A non-invasive marker of disease progression is sorely required. The present study focused on the plasmatic levels of epidermal growth factor receptor (EGFR) in HNSCC patients (N=92) compared with healthy (N=29) and diabetic [type 2 diabetes mellitus (T2DM); N=26] controls. Enzyme-linked immunosorbent assay using antibodies against the extracellular region of EGFR (L25-S645) was performed. No significant changes were observed between diabetic and healthy controls. However, there were significantly higher EGFR plasma levels in HNSCC patients compared with both control groups (P=0.001 and 0.005, respectively). Receiver operating characteristic curve analysis identified a sensitivity of 76.09%, a specificity of 67.27% and an area under curve of 0.727 for this comparison. No significant association was observed between EGFR plasma levels and tumor stage, tumor grade, lymph node or distant metastasis occurrence, smoking habit or hypertension. However, the presence of human papillomavirus infection and T2DM in HNSCC patients had borderline effect on the plasma EGFR levels. Survival analysis revealed no significant influence of plasmatic EGFR levels on the overall and disease-specific survival of HNSCC patients. In conclusion, EGFR plasma levels appear to be a relatively promising diagnostic, but poor prognostic, HNSCC marker.

Fixed combination of cinnarizine and dimenhydrinate versus betahistine dimesylate in the treatment of Ménière's disease: a randomized, double-blind, parallel group clinical study.

In a randomized, double-blind clinical study, we evaluated the efficacy and tolerability of the fixed combination of cinnarizine, 20 mg, and dimenhydrinate, 40 mg (Arlevert [ARL]) in comparison to betahistine dimesylate (12 mg) in 82 patients suffering from Ménière's disease for at least 3 months and showing the characteristic triad of symptoms (paroxysmal vertigo attacks, cochlear hearing loss, and tinnitus). The treatment (one tablet three times daily) extended to 12 weeks, with control visits at 1, 3, 6, and 12 weeks after drug intake. The study demonstrated for both the fixed-combination ARL and for betahistine a highly efficient reduction of vertigo symptoms in the course of the 12 weeks of treatment; however, no statistically significant difference between the two treatment groups could be established. Similar results were found for tinnitus (approximately 60% reduction) and for the associated vegetative symptoms (almost complete disappearance). Vestibulospinal reactions, recorded by means of craniocorpography, also improved distinctly, with a statistically significant superiority of ARL versus betahistine (p < .042) for the parameter of lateral sway (Unterberger's test). The caloric tests (electronystagmography) showed only minor changes for both treatment groups in the course of the study. A statistically significant improvement of hearing function of the affected ear (p = .042) was found for the combination preparation after 12 weeks of treatment. The tolerability was judged by the vast majority of patients (97.5%) in both groups to be very good. Only one patient (betahistine group) reported a nonserious adverse event, and two betahistine patients did not complete the study. In conclusion, the combination preparation proved to be a highly efficient and safe treatment option for Ménière's disease and may be used both in the management of acute episodes and in long-term treatment. Efficacy and safety were found to be similar to the widely used standard therapy with betahistine.

Efficacy and safety of AM-111 in the treatment of acute sensorineural hearing loss: a double-blind, randomized, placebo-controlled phase II study.

OBJECTIVE: To evaluate the efficacy and safety of AM-111, a c-Jun N-terminal Kinase (JNK) ligand, in patients with acute sensorineural hearing loss (ASNHL). STUDY DESIGN: Prospective, double-blind, randomized, placebo-controlled study with follow-up visits on Days 3, 7, 30, and 90. SETTING: Twenty-five European sites (academic tertiary referral centers, private ENT practices). PATIENTS: Approximately 210 patients aged 18 to 61 years presenting within 48 hours after acute acoustic trauma or idiopathic sudden sensorineural hearing loss with mean hearing loss of 30 dB or greater at the 3 most affected contiguous test frequencies. INTERVENTIONS: Single-dose intratympanic injection of AM-111 (0.4 or 2.0 mg/ml) or placebo; optionally, oral prednisolone if hearing improvement was less than 10 dB at Day 7. MAIN OUTCOME MEASURES: Efficacy was assessed by absolute hearing improvement (primary end point, Day 7), percentage hearing improvement, complete hearing recovery, speech discrimination improvement, and complete tinnitus remission. Safety was evaluated by the frequency of clinically relevant hearing deterioration and adverse events. RESULTS: The study failed to demonstrate a treatment benefit for the entire study population because mild-to-moderate ASNHL cases showed unexpectedly strong spontaneous recovery. In severe-to-profound ASNHL patients (threshold ≥60 dB), AM-111 0.4 mg/ml showed statistically significant, clinically relevant, and persistent improvements in hearing and speech discrimination and higher tinnitus remission compared with placebo. The study drug and the intratympanic injections were well tolerated. CONCLUSION: The study established proof of concept for AM-111 in the treatment of severe-to-profound ASNHL. Control for spontaneous hearing recovery is essential for ASNHL studies.

Elevated malondialdehyde correlates with the extent of primary tumor and predicts poor prognosis of oropharyngeal cancer.

The aim of the study was to investigate the relationship between plasma levels of malondialdehyde (MDA), a routinely used marker of oxidative stress, and squamous cell carcinoma of the oral cavity and oropharynx (OSCC). The prospective cohort study comprised a total of 67 patients who underwent surgery for OSCC. MDA was assessed using high performance liquid chromatography. The MDA levels in the pooled T1-2 patients were lower than in the patients with T3-4 tumors. A negative correlation of MDA and tumor grade was shown. Seventeen patients who manifested recurrence during the 49.6 months follow-up had significantly increased MDA compared to those staying in complete remission. Kaplan-Meier analysis revealed that the median disease-free interval and overall survival in the group with MDA > median was 19.3 and 22.5 months respectively, in contrast to 31.5 and 31.6 months respectively, in patients with MDA < or = median. The prognostic value and low cost of MDA measurement could make it a versatile and useful prognostic tool for the identification of OSCC patients with a high risk of recurrence.