Lett Appl Microbiol. 2012 Feb 11;
Amdiouni H, Faouzi A, Fariat N, Hassar M, Soukri A, Nourlil J

Aims:  Reclaimed wastewater is a considerable water resource in Morocco. Its agricultural reuse requires an assessment of viral contamination. The aim of this study was to detect both infectious and non infectious human adenoviruses (HAdV) and enteroviruses (EV) in raw wastewater and treated effluent from wastewater treatment plants (WWTPs) and domestic sewage in Morocco. Methods and Results:  A total of 22 samples were analysed. A Polyethylene glycol (PEG) precipitation method was used, followed by intergrated cell culture -PCR (ICC-PCR) using two cell lines: human rhabdomyosarcoma tumor tissue and laryngeal carcinoma cells (RD and Hep2 cells). Furthermore, viral genome amplification was confirmed by sequencing. HAdV were detected in 10 (45.5%) out of the 22 samples involving two species: HAdV-B and HAdV-D. EV was detected in 5 (23%) samples belonging to Coxsackie B5 virus and Poliovirus vaccine strain (Sabin2). Conclusions:  HAdV and EV were detected in the analysed samples from two WWTPs and HAdV in domestic sewage. Significance and Impact of the Study:  This work is the first study in Morocco using cell culture, PCR and sequencing of enteric viruses from wastewater. The presence of infectious HAdV and EV in treated effluent emphasizes the need of wastewater treatment surveillance. © 2012 The Authors Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

Tumori. 2011 Nov-Dec; 97(6): 781-6
Li R, Wang R, Zhai R, Dong Z

Aim and objective. Laryngeal carcinoma is one of the most aggressive cancers of the head and neck region. The survival rate of patients with laryngeal carcinoma is low due to its late metastases and resistance to chemotherapy and radiotherapy. It was reported that mTOR was involved in the growth and apoptosis of various cancer cells. The aim of this study was to detect the effects of mTOR inhibition by mTOR shRNA on the proliferation, apoptosis and invasive ability of Hep-2 human laryngeal carcinoma cells in vitro. Methods and study design. mTOR shRNA was designed and transfected into Hep-2 human laryngeal carcinoma cells. Untreated cells and cells treated with control vector (non-targeted shRNA) were used as control. The proliferation and apoptosis of Hep-2 cells were detected by MTT and flow cytometry. A transwell assay was used to measure the invasive ability of Hep-2. The inhibition effects on the mTOR signaling pathway by mTOR shRNA were studied using RT-PCR and Western blot.Results. Our results showed that the mRNA and protein expression of mTOR and Akt were high in laryngeal carcinoma cells and could be inhibited by mTOR shRNA. At the same time, low expression of PTEN mRNA and protein was observed in Hep-2 cells. The expression increased when the cells were transfected with mTOR shRNA. This showed that mTOR shRNA could inhibit the proliferation and invasive ability of Hep-2 cells. It also could induce the apoptosis of Hep-2 cells in vitro. Conclusions. The mTOR signaling pathway plays an important role in the development of laryngeal carcinoma. The mTOR shRNA we designed in this experiment effectively inhibited the mTOR signaling pathway. It inhibited the proliferation and invasive ability of the studied laryngeal carcinoma cells and induced their apoptosis in vitro. mTOR might therefore be a useful target in the therapy of laryngeal carcinoma.

Oral Oncol. 2012 Feb 6;
Wang J, Xu Y, Li J, Sun X, Wang LP, Ji WY

Over-expression of DNA methyltransferase 1 (DNMT1) correlates with hypermethylation of tumor suppressor genes (TSGs) in tobacco-induced cancers. The tobacco component nitrosamine 4-(methylnitro-samino)-1-(3-pyridyl)-1-butanone (NNK) increases protein levels of the DNMT1 in human lung cancer. However, the role of DNMT1 expression induced by NNK is not clear during laryngeal carcinogenesis. We investigated DNMT1 expression levels in 101 cases of human laryngeal carcinoma specimens and 54 cases clear surgical margin specimens by reverse transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry (IHC) detection. Then, we analyzed the relationship between the DNMT1 expression and the smoking status of the patients with laryngeal carcinoma. Moreover, we investigated the effects of tobacco carcinogen NNK on DNMT1 expression in Hep-2 cells. We found that DNMT1 mRNA and protein expressions were up-regulated in laryngeal cancer tissues (p<0.05 and p<0.01, respectively). Among the 101 cases, DNMT1 protein from patients with heavier smoking habit had a significant trend of an increase with IHC scores (p<0.01). The overall survival rates of patients DNMT1-positive were significantly lower than those of patients DNMT1-negative (p<0.05). We observed that NNK increased DNMT1 protein levels, not mRNA levels, in cultured Hep-2 cells significantly in both dose- and time-dependent manner (p<0.05). These results supported the idea that NNK-induced DNMT1 expression may result from protein stabilization. Increased DNMT1 protein expression may play a critical role in the malignant progression of larynx.

Tumour Biol. 2012 Feb 7;
Zeng YC, Xue M, Chi F, Xu ZG, Fan GL, Wu R, Fan YC, Zhong WZ, Wang SL, Zhang XY, Wu LN, Chen XD, Jin XY, Duan QY, Xu R, Chen W, Qian HC, Xiao YP

The aim of this study was to investigate the effect of C-reactive protein (CRP) level on the prognosis of patients with locoregionally advanced laryngeal carcinoma treated with chemoradiotherapy. Fifty-seven patients with locoregionally advanced laryngeal carcinoma (cT3-4, N0-3, M0) treated with chemoradiotherapy were reviewed retrospectively. Chemoradiotherapy comprised external beam radiotherapy to the larynx (70 Gy) with three cycles of cisplatin at 3-week intervals. Elevated CRP was defined as >8 mg/L. The survival rate was calculated using the Kaplan-Meier method, and a multivariate analysis was used to identify significant factors associated with prognosis, using a Cox proportional hazards model. During the median (range) follow-up of 5 years (1.3-5), 29 patients died from laryngeal cancer; the 5-year cancer-specific survival (CSS) rate was 49.12%. Fifteen patients had a high CRP level before chemoradiotherapy (>8 mg/L), and their CSS rate was significantly worse than that in the remaining patients (P = 0.003). Multivariate analysis showed that CRP and tumor site were independent prognostic indicators for CSS, with a hazard ratio of 2.66 (95% confidence interval (CI), 1.22-5.82; P = 0.014) and a hazard ratio of 1.67 (95% CI, 1.01-2.77; P = 0.045), respectively. Of those with elevated CRP, the CRP levels of ten patients became normal after chemoradiotherapy, of whom four were alive with no evidence of recurrence or metastasis during the follow-up. By contrast, all six with no CRP normalization after chemoradiotherapy died within 3.8 years. The elevation of CRP before treatment predicts a poor prognosis in patients with locoregionally advanced laryngeal carcinoma treated with chemoradiotherapy.

Eur Arch Otorhinolaryngol. 2012 Feb 5;
Hassumi-Fukasawa MK, Miranda-Camargo FA, Guimarães MC, Simões RT, Donadi EA, Soares CP, Soares EG

Laryngeal squamous cell carcinoma is one of the most common malignant neoplasms of the head and neck. In Brazil, laryngeal tumors represent 2% of all cancers and are associated with approximately 3,000 deaths annually. Human papillomavirus (HPV) has been reported to play an important role in the etiology of laryngeal cancer. The aim of the present study was to evaluate the expression of p53, p27, and Mdm2 in laryngeal carcinomas. Sixty-three larynx biopsies were selected for the study, including 9 in situ laryngeal carcinomas, 27 laryngeal carcinomas without metastasis and 27 laryngeal carcinomas with metastasis. Twenty-seven cervical lymph nodes from patients with metastatic lesions were also evaluated. The expression levels of p53, p27, and Mdm2 were assessed by immunohistochemistry using a computer-assisted system. HPV detection and typing were performed using PCR, and the HPV types that were evaluated included HPV 6, 11, 16, 18, 31 and 33. Out of 63 patients, 53 (84.1%) were positive for β-globin (internal control), and 10 (15.9%) were β-globin negative and therefore excluded from the evaluation. Thus, 7 (13.2%) out of 53 patients were HPV positive, and 46 (86.8%) out of 53 patients were HPV negative. Statistically significant differences (p < 0.05) in Mdm2 expression levels were observed in the in situ laryngeal carcinoma samples compared with the laryngeal carcinoma samples with metastasis. No statistically significant differences (p > 0.05) in either p53 or p27 expression levels were detected. These findings suggest that Mdm2 may be associated with the invasiveness and aggressiveness of laryngeal carcinomas.

FEBS Lett. 2012 Feb 1;
Bian K, Fan J, Zhang X, Yang XW, Zhu HY, Wang L, Sun JY, Meng YL, Cui PC, Cheng SY, Zhang J, Zhao J, Yang AG, Zhang R

Previous studies have shown that miR-203 acts as a tumor-suppressive microRNA in various cancers, but its roles in laryngeal carcinoma are still contradicted. Here, we found that miR-203 inhibited the growth of laryngeal cancer cells and survivin was a direct target of miR-203. Moreover, silencing of survivin recapitulated the effect of miR-203 on cell cycle progression, whereas overexpression of survivin reversed this effect. Additionally, qRT-PCR showed the reciprocal relationship between miR-203 and survivin in laryngeal cancer tissues. These findings indicate that miR-203 inhibits the proliferation of laryngeal carcinoma cells by directly targeting survivin, suggesting its application in anti-cancer therapeutics.

Southeast Asian J Trop Med Public Health. 2011 Sep; 42(5): 1072-6
Larbcharoensub N, Cheewaruangroj W, Nitiyanant P

Laryngeal sarcocystosis is an uncommon zoonotic coccidian protozoal infestation of human beings. The authors reviewed the pathology of 1,063 laryngeal biopsies over the past 10 years (2000 to 2009). Only one case of laryngeal sarcocystosis accompanying laryngeal squamous cell carcinoma was identified. The overall prevalence of laryngeal sarcocystosis was 0.094%. The case was a 66-year-old man who presented with voice hoarseness for six months. Physical examination and computed tomography revealed an ulcerative exophytic mass on the right true vocal cord, suggestive of laryngeal carcinoma. He underwent a right frontolateral partial laryngectomy. Histopathology showed a nonkeratinizing squamous cell carcinoma with Sarcocystis spp in the vocalis muscle. He was followed up and enrolled in speech therapy. The authors briefly review the clinicopathologic features and pathogenesis of muscular sarcocystosis and concurrent laryngeal sarcocystosis and squamous cell carcinoma.

Arch Med Sci. 2011 Jun; 7(3): 517-22
Klatka J, Remer M, Dobrowolski R, Pietruszewska W, Trojanowska A, Siwiec H, Charytanowicz M

The aim of the study was to determine the content of cadmium (Cd), cobalt (Co) and nickel (Ni) in the samples from laryngeal carcinoma in comparison with the level of these elements in the samples of healthy mucous membrane from the same larynx.The study was conducted on 43 patients with laryngeal carcinoma. The levels of Cd, Co, and Ni in carcinoma and healthy control tissues was determined by inductively coupled plasma optical emission spectrometry (ICP - OES) using sequential spectrometer.No significant differences were found between the levels of the Cd, Co, and Ni in laryngeal carcinoma vs tissues without carcinoma. However, it was noted that the concentration of Cd in tumors of patients with metastases to cervical lymph nodes was significantly higher than in tumors without metastases. The content of Co was significantly higher in more advanced laryngeal tumors: in stage-T4 than in stage T3. It is of interest that the levels of Cd, Co and Ni were significantly higher in tumors in patients from rural than urban areas.The imbalance in the level of nickel, cadmium and cobalt in laryngeal cancer may be due to a changed cellular metabolism in the cancer process. However, the results of our study reveal the significant differences in the concentration of these metals between patients from urban and rural areas which suggests that this fact may be related to environmental or occupational factors and therefore it requires further study.

Bull Acad Natl Med. 2011 Mar; 195(3): 741-53
Laccourreye O, Garcia D, Maldent JB, Werner A

Based on a comparison of two cohorts of patients with laryngeal cancer managed by laryngectomy one century apart, and an analysis of original textbooks from the end of the nineteenth century, we examine the development of laryngectomy, the difficulties encountered in this first therapeutic approach to laryngeal carcinoma, and the conflicts it generated. The overall death rate changed little over the course of a century, although the 5-year actuarial survival estimate improved from 22.6% in 1888 to 75.1% in 1988 (p < .0001). The causes of death also changed (p < .0001), with fewer deaths related to post-operative complications and/or local failure (82.7% in 1888, 9% in 1988). The rate of suicide deaths fell from 1.8% to 0.04%. In contrast, the percentage of deaths due to metachronous second primary tumors and/or intercurrent disease increased from 9% in 1888 to 76.7% in 1998. Analysis of actuarial survival rates demonstrated that 77.4% of patients died before the 60th post-operative month in 1888, while this percentage was only reached 275 months post-operatively a century later. Our research also underscores the importance of patient information and medical ethics.

Oral Oncol. 2012 Jan 27;
Marioni G, Staffieri A, Lora L, Fermo S, Giacomelli L, La Torre FB, Favaretto N, Valentini E, Manzato E, Blandamura S

Cancer is common in the elderly, who may also be frail, which can complicate the choice of the best therapeutic approach. We sought to examine whether the serine-threonine kinase mTOR, a "master switch" in cancer cells that modulates metabolism, the cell cycle, and apoptosis, might help in clinical decision-making. The aim of the present study was thus to assess the potential prognostic role of mTOR in elderly patients with laryngeal carcinoma (LSCC). mTOR expression was determined immunohistochemically in 54 consecutive elderly (⩾65years old) patients with LSCC. On univariate analysis, nodal involvement and pathological stage correlated strongly with the elderly LSCC patients' prognosis in terms of disease recurrence rate and disease-free survival (DFS). Patients whose mTOR expression was >35.3% had a significantly higher recurrence rate (p=0.003) and shorter DFS (p=0.013). In the multivariate model, N status (p=0.001) and mTOR expression (p=0.026) maintained an independent prognostic significance in relation to DFS. mTOR probably influences the aggressive LSCC phenotype in elderly patients and its expression in elderly LSCC cases can be considered a prognostic marker potentially useful for identifying patients at higher risk of disease recurrence, and N0 patients at higher risk of recurrence who may benefit from more aggressive treatment. Since rapalogs (as mTOR inhibitors) might have an effect on LSCC, further investigations are needed to ascertain these agents' role in therapeutic strategies for advanced LSCC in elderly patients.

HNO. 2012 Jan; 60(1): 41-3
Schroeder U, Gebhard MP, Wollenberg B

Although the glottis is amenable to chemotherapy, currently most lesions from stage I laryngeal dysplasia up to carcinoma in situ are excised. This literature review presents selected molecular biological aspects especially in relation to dysplasia of the larynx and its therapy, as well as currently preferred biomarkers for chemotherapeutic prevention of laryngeal dysplasia.

HNO. 2012 Jan; 60(1): 32-40
Pantel M, Guntinas-Lichius O

Laryngeal cancer is one of the most frequent types of head and neck cancer. The incidence is decreasing for men but still increasing for women. The incidence in Germany is about 5-7/100,000 persons/year for men and 0.6-0.8/100,000 persons/year for women. Due to the increased life expectancy, the average age of patients diagnosed with laryngeal cancer is increasing. Nevertheless, adequately prepared older patients treated by standard protocols can have the same survival and complication rates as younger patients. Tobacco and alcohol are still the primary risk factors responsible for disease in at least 80% of the patients. Despite the many new diagnostic tools, still more than half of the patients are diagnosed at an advanced tumor stage. Survival rates have not improved significantly in the last 10 years in Germany, and the average 5-year overall survival rate is about 60%. However, a decrease in the survival rate, as observed in the USA, cannot be confirmed for Germany.