Hyaluronan synthases and hyaluronidases in nasal polyps.

Nasal polyps (NPs) are benign lesions of nasal and paranasal sinuses mucosa affecting 1-4 % of all adults. Nasal polyposis affects the quality of patient's life as it causes nasal obstruction, postnasal drainage, purulent nasal discharge, hyposmia or anosmia, chronic sinusitis, facial pain and snoring. Without treatment, the disease can alter the craniofacial skeleton in cases of extended growth of polyps. The development of NPs is caused by the hyperplasia of nasal or paranasal sinuses mucosa, and edema of extracellular matrix. This is usually the result of high concentration of high molecular mass hyaluronan (HA) which is either overproduced or accumulated from blood supply. The size of HA presents high diversity and, especially in pathologic conditions, chains of low molecular mass can be observed. In NPs, chains of about 200 kDa have been identified and considered to be responsible for the inflammation. The purpose of the present study was the investigation, in NPs and normal nasal mucosa (NM), of the expression of the wild-type and alternatively spliced forms of hyaluronidases, their immunolocalization, and the expression of HA synthases to examine the isoform(s) responsible for the increased amounts of HA in NPs. Hyaluronidases' presence was examined on mRNA (RT-PCR analysis) and protein (immunohistochemistry) levels. Hyaluronan synthases' presence was examined on mRNA levels. Hyaluronidases were localized in the cytoplasm of epithelial and inflammatory cells, as well as in the matrix. On mRNA level, it was found that hyal-1-wt was decreased in NPs compared to NM and hyal-1-v3, -v4 and -v5 were substantially increased. Moreover, HAS2 and HAS3 were the only hyaluronan synthases detected, the expression of which was almost similar in NPs and NM. Overall, the results of the present study support that hyaluronidases are the main enzymes responsible for the decreased size of hyaluronan observed in NPs; thus they behave as inflammatory agents. Therefore, they could be a potential target for the design of a more advanced treatment for nasal polyposis.

Determination of matrix metalloproteinase 9 (MMP-9) protein expression in laryngeal squamous cell carcinomas based on digital image analysis.

PURPOSE: Matrix metalloproteinases (MMPs) is a superfamily of proteins involved in angiogenesis and metastatic tissue invasion in many cancers. Overexpression of MMP- 9 has been detected in significant proportions of laryngeal squamous cell carcinomas (LSCCs), but its prognostic impact remains unclear. In this study we performed a digital image analysis for analyzing MMP-9 protein expression in a series of LSCCs correlating them with clinicopathological factors. METHODS: MMP-9 protein expression level was determined immunohistochemically in 30 tissue sections surgically derived from patients (21 male and 9 female) with LSCC. Using digital image analysis, we measured their corresponding protein expression levels (staining intensity/S.I. range values 0-255). RESULTS: Moderate and high MMP-9 protein expression levels (grouping as 2+/3+ overexpression) were detected in 19/30 (63.3%) cases. Statistical significance was observed correlating stage with SI (p=0.02), whereas a borderline association with differentiation grade of the examined tumors was also registered (p=0.05). Interestingly, high levels of MMP-9 expression were observed in cases that demonstrated a significant level of inflammatory (predominantly lymphocytic) infiltration. CONCLUSION: MMP-9 protein overactivation is a frequent and significant genetic event in LSCC, correlating with its biological behavior (increased TNM stage). MMP-9 seems to mediate an epithelial-stromal intra-reaction correlating also with induction of specific inflammation pathways.

MIF attenuates the suppressive effect of dexamethasone on IL-6 production by nasal polyp.

BACKGROUND: Nasal polyposis (NP) is a chronic inflammatory disease of the upper airways, that characterized by inflammatory cells infiltration, extracellular matrix accumulation and oedema. Interleukin-6 (IL-6) is a multifunctional cytokine, implicated in various inflammatory conditions, including NP pathogenesis. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory mediator able to antagonize the inhibitory effects of glucocorticoids on the expression of various cytokines and growth factors. AIM: To investigate the presence of MIF in nasal polyp tissues and the influence of a MIF activity inhibitor on dexamethasone effects on IL-6 production. PATIENTS AND METHODS: Nasal polyps were resected by functional endoscopic sinus surgery for treatment of chronic sinusitis with polyposis and healthy nasal mucosa was taken during nasal septoplasty-chochoplasty. MIF and IL-6 levels were determined by ELISA. The expression of MIF and IL-6 at the mRNA level was ascertained by RT-PCR. RESULTS: MIF was detected in all polyp tissue extracts and tissue cultures conditioned media. MIF and IL-6 expression were significantly higher in polyp tissues as compared to normal nasal mucosa tissues. Dexamethasone at concentration 1-100 microM caused a statistically significant dose-dependent suppression of IL-6 production by polyp tissue cultures. Inhibition of MIF by (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1), an inhibitor of MIF tautomerase activity, significantly enhanced the dexamethasone suppressive effect on IL-6 production. CONCLUSIONS: MIF, presence in polyp tissue, attenuates the suppressive effect of dexamethasone on the production of IL-6 by this tissue, since the simultaneous use of its inhibitor ISO-1 leads to an enhancement of dexamethasone activity. Therefore, it is reasonable to propose that the utilization of MIF inhibitors together with glucocorticoids in clinical practice may be beneficial in the treatment of NP.

Presence of hyaluronidase isoforms in nasal polyps.

BACKGROUND: Nasal polyps are benign lesions originating from the nasal mucosa or paranasal sinuses. The most important etiological factor seems to be increased hydration of epithelium and hyperplasia of the extracellular matrix, which may involve hyaluronan, a high molecular mass extracellular glycosaminoglycan. Degradation of hyaluronan proceeds through the action of specific hyaluronidases. OBJECTIVE: The aim of the present study was to investigate the hydrodynamic size of hyaluronan and the presence of the various hyaluronidase isoforms in nasal polyps. METHODS: Samples of polypoid mucosal tissue and normal nasal mucosa were obtained from twenty patients suffering from nasal polyposis. Zymographic analysis and western blotting were used to detect hyaluronidase activity. RESULTS: The results indicated the presence of hyaluronan of small molecular mass in all samples examined. About one third of it has a mean molecular mass of 240 kDa, exactly that required for the expression of inflammatory response. Laboratory analysis suggested that degradation of hyaluronan occurred through the action of three hyaluronidase isoforms: Hyal-1, Hyal-2 and PH-20. CONCLUSIONS: Since hyaluronan fragments of 200-250 kDa induce the expression of inflammatory cytokines, a specific role of hyaluronidases in the development or progression of nasal polyps may be concluded. Therefore, new treatment protocols may be proposed.

Expression and distribution of aggrecanases in human larynx: ADAMTS-5/aggrecanase-2 is the main aggrecanase in laryngeal carcinoma.

Members of the ADAMTS family of proteases degrade proteoglycans and thereby have the potential to alter tissue architecture and regulate cellular functions. Aggrecanases are the main enzymes responsible for aggrecan degradation, due to their specific cleavage pattern. In this study, the expression status, the macromolecular organization and localization of ADAMTS-1, ADAMTS-4/aggrecanase-1 and ADAMTS-5/aggrecanase-2 in human normal larynx and laryngeal squamous cell carcinoma (LSCC) were investigated. On mRNA level, the results showed that ADAMTS-4 was the highest expressed enzyme in normal larynx, whereas ADAMTS-5 was the main aggrecanase in LSCC presenting a stage-related increase up to stage III (8-fold higher expression compared to normal), and thereafter decreased in stage IV. Accordingly, immunohistochemical analysis showed that ADAMTS-5, but not ADAMTS-4, was highly expressed by carcinoma cells. Sequential extraction revealed an altered distribution and organization of multiple molecular forms (latent, activated and fragmented forms) of the enzymes within the cancerous and their corresponding macroscopically normal laryngeal tissues, compared to the normal ones. Importantly, these analyses indicated that critical macromolecular changes occurred from the earliest LSCC stages not only in malignant parts of the tissue but also in areas that were not in proximity to carcinoma cells and appeared otherwise normal. Overall, the results of the present study show that ADAMTS-5/aggrecanase-2 is the main aggrecanase present in laryngeal carcinoma suggesting a critical role for the enzyme in aggrecan degradation and laryngeal tissue destruction during tumor progression.

Factors influencing quality of life after total laryngectomy: a study of 92 patients.

BACKGROUND AND OBJECTIVES: Total laryngectomy is a radical surgery for advanced laryngeal cancer. In the present study we evaluate the influencing factors for long term quality of life of 92 laryngectomees. PATIENTS AND METHODS: 92 patients who underwent laryngectomy for laryngeal cancer in the University Hospital of Patras, Greece were evaluated using a modified version of the EORTC-QOL-C30 questionnaire. The patients were distributed into 6 groups based on time elapsed between surgical treatment and completion of the questionnaire, ranging from 2 years for group 1 to 9-10 years for group 6. RESULTS: The mean value of psychological status (p=0.01), smelling disorders (p=0.032), mood change (p=0.003) were statistically significant in regard to the patient's overall view of their life postsurgically. Coexisting illness (defined as diabetes, hypertension or prostate disease which required medication on daily basis or hospitalization more than two times a year) was also significantly affecting (p=0.021) the patient's oral communication skills. CONCLUSION: Quality of life is affected by functional disabilities and the psychological state of the patients. No statistically significant improvement over time was noted in our series.

Necrotizing fasciitis of the head and neck: report of 5 cases and review of the literature.

BACKGROUND: Necrotizing fasciitis (NF) is a severe, rapidly spreading soft tissue infection of polymicrobial origin. This condition most frequently affects thorax, abdominal wall, extremities, perineum and groin, but according to recent literature the head and neck area is also involved with increasing frequency. PATIENTS AND METHODS: Five cases of head and neck NF were detected among patients who were admitted at the Department of Otorhinolaryngology-Head and Neck Surgery of the University Hospital of Patras, Patras, Greece, over a 5-years period. Various parameters including patients' health status, co-morbidity, etiology, microbiology, affected area, antibiotic therapy, hospital stay, surgical treatment and complications were considered. CONCLUSIONS: The management of NF should comprise of hemodynamic and respiratory evaluation and monitoring, broad-spectrum i.v. antimicrobial therapy, surgical debridement and nutritional support. Close postoperative management of NF patients remains of paramount importance.

Sinonasal sarcoidosis. A case report.

INTRODUCTION: Sarcoidosis is a granulomatous disease of unknown etiology, which in the majority of cases affects the lower respiratory tract. Although neck mass, parotid swelling and facial nerve palsy are considered the primary complaints in cases of head and neck sarcoidosis, sinonasal localization of the disease is also uncommonly reported. CASE: A 46 year old woman presented in our clinic with complaints of nasal congestion over a six month period. The patient reported increasing symptom severity in the last month with occasional purulent discharge. No other symptoms were reported. The patient's medical record was significant only for sarcoidosis localized to the lower respiratory tract. On anterior rhinoscopy, the nasal mucosa presented marked hypertrophy, purplish discoloration and granulomatous appearance of the inferior turbinates on the left side. The diagnosis of sinonasal sarcoidosis was made based on histopathologic examination of biopsy specimens. CONCLUSION: Sinonasal sarcoidosis presents a challenging issue for the clinicial as its mimicking features may be misleading and delay diagnosis. In the present paper we present a case of sinonasal sarcoidosis presenting as chronic rhinosinusitis.

Voice restoration after total laryngectomy using provox 2 (generation II) prosthesis.

BACKGROUND AND OBJECTIVES: Voice restoration after total laryngectomy is of outmost importance for patient and a therapeutic challenge for the surgeon and speech pathologist. Among various rehabilitation methods prosthetic voice yields nowadays the best results. PATIENTS AND METHODS: A total of 12 patients underwent laryngectomy at the Department of Otorhinolaryngology, University Hospital, Patras, Greece, between February 2006 and May 2007. All patients had advanced laryngeal squamous cell carcinoma. Eight patients had primary voice prosthesis inserted and four patients had a tracheo-esophageal puncture (TEP) carried out as a secondary procedure. RESULTS: The majority of the patients (80%) from both the primary insertion or the secondary insertion group, developed good and understood speech using the prosthesis. No significant difference in quality of speech was found between the two subgroups. Four patients required replacement of the prosthesis at intervals of 8 and 10 months after insertion, because of salivary leakage and granulation formation around the fistula, while 3 patients developed a mild mucositis and tracheitis due to postoperative radiotherapy. DISCUSSION: TEP puncture and prosthesis insertion is a relatively simple, safe and effective surgical procedure for voice restoration after laryngectomy. Provox 2 (generation II) voice prosthesis is a new and useful modified device that has overcome the previous prosthesis drawbacks, is well tolerated by the patients and can be easily changed via the front-loading technique.

Stage-related decorin and versican expression in human laryngeal cancer.

BACKGROUND: The major proteoglycan of normal human larynx is aggrecan. In laryngeal carcinoma, aggrecan is depleted, with versican and decorin appearing in higher amounts. MATERIALS AND METHODS: Proteoglycans in laryngeal carcinoma samples were characterized immunohistochemically and using Western blotting; their expression was examined by RT-PCR. RESULTS: Aggrecan was totally removed in advanced cancer and its RT-PCR product was not identified. Both versican and decorin were overexpressed in cancer, versican much more than decorin. Decorin expression was higher than that of versican in the normal larynx; therefore, their disproportionate overexpression during cancer resulted in about equimolar expression. Both proteoglycans' expression correlated with their stage-related accumulation within the tissue. CONCLUSION: These data add to our previous findings and support the view that the levels of expression and the extent of accumulation and localization in the tumor stroma of structurally modified versican and decorin could be associated with the degree of aggressiveness of laryngeal carcinoma.

Concurrent tonsillitis and pneumonia due to S. pyogenes.

Tonsillitis is a common disease especially of children and young adults. The diagnosis of tonsillitis generally requires the consideration of Group A beta-hemolytic streptococci infection. Pneumonia is an inflammation of the alveoli and terminal airspaces in response to invasion by an infectious agent that is introduced into the lungs through hematogenous spread or inhalation. We present a case of a concurrent acute tonsillitis and pneumonia due to S. pyogenes. To our knowledge, a similar case has not been previously described in the literature.

Palatine tonsillar metastasis from lung adenocarcinoma.

Tonsillar metastasis from lung cancer is an extremely rare event and is usually related to metastatic spread of the neoplastic disease to other organs with poor prognosis. We present the unusual case of a 71-year-old man who developed a huge metastasis to the left palatine tonsil from a lung adenocarcinoma. The tumor was exophytic with necrotic and hemorrhagic areas, occupying a large portion of the oropharynx and producing airway obstruction, thus necessitating tracheostomy. Radiation therapy was delivered in an attempt to reduce the size of the tumor. Although a mild decrease of the tonsillar mass was noted, the designed therapy had to be discontinued because of a rapid deterioration of the patient's general condition. The patient finally died of disseminated disease. This case highlights the need for a thorough evaluation of a suspicious tonsillar enlargement, as this may be a sign of a primary malignancy elsewhere in the body.

Expression of metalloproteinases and their tissue inhibitors in squamous cell laryngeal carcinoma.

The hallmark of cancer invasion is the degradation of extracellular matrix components. Matrix metalloproteinases are the major enzymes participating in this event and their activity is regulated extracellularly by their presence as proenzymes and the concomitant presence of the specific tissue inhibitors. The present study describes the immunohistochemical localization of gelatinases, matrix metalloproteinase (MMP)-2 and -9 and tissue inhibitor of metalloproteinase (TIMP)-1 and -2 in human laryngeal carcinoma and their expression with respect to tumor classification and compared with the respective healthy subjects. MMP-2 was immunolocalized in the cytoplasm of the epithelial cells and in the loose connective tissue, whereas MMP-9 was also observed in basement membrane and chondrocytes. Both were also found in tumor cells, but staining was decreased with increasing stage of cancer. TIMP-1 was present exclusively in stroma and totally absent from tumor cells and it was overexpressed in normal cells surrounding the tumor. TIMP-2 was identified in the cytoplasm of epithelial cells, in stroma and sometimes in chondrocytes. In addition, it was present in tumor cells of only stage IV samples. The expression level of both gelatinases and TIMPs increased as the stage of cancer increased, suggesting the possible post-transcriptional removal of their mRNA. These observations, performed in a given head and neck site, suggest that the behavior of head and neck tumors seems to depend on the site and additional studies should be performed to obtain a general understanding of the disease and ascertain the role of the constituents examined.

Hyaluronidase and CD44 hyaluronan receptor expression in squamous cell laryngeal carcinoma.

Squamous cell laryngeal carcinoma undergoes significant structural-related modifications of the extracellular matrix components (ECM), the most characteristics being the presence of degraded collagen, aggrecan and hyaluronan. We examined the presence of hyaluronidase and of the cellular hyaluronan receptor CD44 during the various stages of cancer. ECM components were extracted by using PBS, 4 M GdnHCl and 4 M GdnHCl-0.1% Triton-X 100 sequentially and hyaluronidase and CD44 analyzed by zymography and immunochemistry techniques. Total RNA was also extracted and the mRNA of the various hyaluronidases and of CD44 was analyzed after amplification with RT-PCR. Hyaluronidase was detected as a double band of 45 and 55 kDa molecular mass, only in cancer samples. The analysis of mRNA indicated an aberrant expression of PH-20, the testicular-type hyaluronidase, at late stages of cancer and an overexpression of HYAL1 only at stage IV. In addition, CD44 was identified in two protein bands of 80 and 64 kDa in cancer samples. The analysis of mRNA showed that hyaluronan receptor was expressed in a stage-related order. Thus, it could be suggested that in laryngeal squamous cell carcinoma, cancer cells migrated and proliferated under the influence of small molecular mass hyaluronan, by expressing increased amounts of its receptor.

The management of squamous cell carcinoma of the neck. The Birmingham UK experience.

AIMS: To review our treatment strategy and outcomes for metastatic squamous cell carcinoma of the neck. METHODS: One hundred and six consecutive patients treated between 1992 and 1998 were analysed retrospectively. The following data were obtained. Demographic details, tumour site, clinical and pathological TMN staging, tumour grade and presence of extracapsular spread, treatment modality (surgery, radiotherapy and chemotherapy), type of neck dissection and complications, 2-year loco-regional control and 5-year overall survival. RESULTS: Ninety-two patients had advanced disease (stages 3 and 4) and of these, 57% had palpable neck metastases. One hundred and six patients underwent a total of 132 neck dissections. Seventy-three patients had post-operative radiotherapy to both sides of the neck and a total of 31 patients took part in the UKHAN 1 trial. Seventy percent of patients achieved 2-year loco-regional control and 63% survived 5-years. CONCLUSION: Metastatic squamous cell carcinoma of the neck can successfully be treated with an aggressive surgical approach and post-operative radiotherapy when indicated. Excellent 2-year loco-regional control and 5-year survival rates are possible.

HPLC identification of oversulphated chondroitin as the major glycosaminoglycan of human tonsils.

The glycosaminoglycans of human nasopharyngeal and palatine tonsils, obtained after surgical dissection due to tonsillitis, were isolated and characterized by means of enzyme susceptibility and HPLC. Chondroitin/dermatan sulphate were the major glycosaminoglycans identified. A large proportion of this glycosaminoglycan was made up of oversulphated structures, namely DeltaDi-di(4,6)S, which were found mainly in invertebrate tissues and in mast cells.

Diagnostic and classification value of metalloproteinases in squamous human laryngeal carcinoma.

Metalloproteinases (MMPs) are a class of enzymes largely involved in tumour progression and metastasis. At least twenty different enzymes are recognized that are also present under normal state of tissues. Their activity is regulated by their presence as proenzymes and by the concomitant presence of the respective tissue inhibitors (TIMPs). The present study describes the alterations of MMPs observed in human laryngeal carcinoma with respect to tumour classification and compares their activity in normal and cancerous tissues and biopsy specimens. Samples from five patients who underwent laryngectomy, from five biopsies and three from autopsies were used. The MMPs of normal and malignant human laryngeal cartilage and of biopsy specimens were identified immunochemically and by zymography using gelatin or casein as substrates. Healthy cartilage from autopsies was found to contain almost exclusively MMP-1, proMMP-2 and proMMP-9. Normal parts from laryngectomies contained, in addition, significant amounts of active MMP-2. The respective malignant parts contained both MMP-2 and -9 in increased amounts in their latent and active forms. Similar profile of MMPs was also identified in tissues surrounding affected cartilage. These alterations were found to be in good accordance with tumour stage and were also observed in biopsy samples. Thus, analysis of MMPs in biopsies can be used together with the clinicopathological parameters for the classification or early diagnosis of laryngeal tumours.

Alterations in the content and composition of glycosaminoglycans in human laryngeal carcinoma.

Glycosaminoglycans in normal and cancerous human laryngeal cartilage were isolated and characterized by means of enzyme susceptibility and high performance liquid chromatography. The known mammalian glycosaminoglycans were identified in all samples but their content and composition varied between normal and malignant samples. Chondroitin/dermatan sulphate was the major glycosaminoglycan in all cases, but its relative proportion was decreased in malignant samples. Its sulphation pattern showed that in normal samples it was sulphated mainly at the C6 position of galactosamine, whereas in malignant samples it was sulphated mainly at C4. Dermatan sulphate, expressed as a result of the different digestion of samples with chondroitinases, was present in very small amounts in normal samples (2.7% of total sulphated glycosaminoglycans) but increased in proportion up to 27.7% in malignant samples. The content of oversulphated chondroitin/dermatan was increased twofold in malignant samples. The content of heparan sulphate was increased almost fivefold in malignant samples as compared to normal ones. The content of hyaluronan was increased in malignant samples 3.5-fold, amounting to up to 11.4% of total glycosaminoglycans. These dramatic changes in the content and composition of glycosaminoglycans seemed to be characteristic of the tumour and independent of its status.

The role of nasogastric tube after elective abdominal surgery.

PURPOSE: Nasogastric tube (NGT) is routine employed after abdominal surgery still in our country and abroad too, but the patients' discomfort is severe and several serious complications were referred as related to its use. The aim of this study was to evaluate the pros and cons of routine use of NGT in elective uncomplicated abdominal surgery. PATIENTS AND METHODS: In order to evaluate the routine use of NGT we performed a prospective randomized trial on 100 patients who had elective uncomplicated abdominal surgery: 50 subjects had the early removal of NGT and in 50 patients it was maintained until passage of flatus/feces. RESULTS: The relevant differences between the two groups were the earlier passage of flatus and feces and the lower incidence of postoperative gastrectasy in the group where NGT was early removed. No statistical difference was recorded concerning the occurrence of postoperative nausea, vomiting, abdominal distension, complications and day of clinical release. CONCLUSIONS: The routine application of NGT in elective abdominal surgery could be omitted if the surgeon and nursing team are willing to renounce a useful tool in providing informations about the resolution of postoperative intestinal atony to the patient's benefit: in fact a severe discomfort due to the NGT was recorded in the 70% of our series.