Quantitative expression analysis of the apoptosis-related gene, BCL2L12, in head and neck squamous cell carcinoma.

BACKGROUND: BCL2L12 is a recently identified gene belonging to the BCL2 family, members of which are implicated in head and neck squamous cell carcinoma (HNSCC). We have recently shown that BCL2L12 mRNA expression is an unfavorable prognostic indicator in nasopharyngeal carcinoma (NPC) and that BCL2L12 can be regarded as a novel, useful tissue biomarker for the prediction of NPC patients' short-term relapse. The aim of this study was to analyze the mRNA expression of the novel apoptosis-related gene BCL2L12 in patients with HNSCC. METHODS: Total RNA was isolated from 53 malignant tumors originating in larynx, pharynx, tongue, buccal mucosa, parotid glands, and nasal cavity, as well as from 34 adjacent non-cancerous tissue specimens, resected from patients with HNSCC. A highly sensitive real-time PCR method for BCL2L12 mRNA quantification in head and neck tissues was developed using the SYBR(®) Green chemistry. After preparing cDNA by reverse transcription, relative quantification was performed using the comparative C(T) () method. RESULTS: BCL2L12 mRNA levels were lower in laryngeal tumors of advanced tumor, node, metastasis (TNM) stage or bigger size and in well-differentiated malignant tongue neoplasms, compared with early-stage laryngeal tumors or poorly differentiated tongue tumors. Interestingly, the BCL2L12 expression showed significant discriminatory value, distinguishing efficiently patients with tongue squamous cell carcinoma (SCC) from non-cancerous population. CONCLUSIONS: This is the first study examining the BCL2L12 mRNA expression in HNSCC. Our results suggest that BCL2L12 mRNA expression may serve as a potential prognostic biomarker in tongue and/or larynx SCC, which principally constitute the great majority of HNSCC cases worldwide.

Nasal chondrogenic tumors: Case reports of primary chondroma and recurrence of atypical cartilaginous tumor (chondrosarcoma grade I) with review of the literature.

Nasal chondromas and nasal atypical cartilaginous tumors (or chondrosarcoma grade 1) are very rare chondrogenic tumors. Approximately 150 cases of nasal chondroma and fewer than 50 cases of nasal atypical cartilaginous tumors have been described. We reported a case with nasal chondroma in the right ethmoid area and a case with a nasal atypical cartilaginous tumor of the anterior septum. No chondroma recurrence was noted for 17 years after endoscopic resection. Recurrence was noted for the nasal atypical cartilaginous tumor 36 months after wide surgical excision via midfacial degloving. Revision surgery was performed again via midfacial degloving. Recurrences of nasal chondroma and atypical cartilaginous tumors are extremely rare. The current data indicate adequate surgical excision, in some cases even radical, as the most important parameter to avoid recurrence. For inoperable recurrent cases of atypical cartilaginous tumors, observation may be followed.

L-DOPA decarboxylase mRNA expression is associated with tumor stage and size in head and neck squamous cell carcinoma: a retrospective cohort study.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) represents one of the most commonly diagnosed malignancies worldwide. The DDC gene encodes L-DOPA decarboxylase, an enzyme catalyzing the decarboxylation of L-DOPA to dopamine. We have recently shown that DDC mRNA is a significant predictor of patients' prognosis in colorectal adenocarcinoma and prostate cancer. The aim of the current study was to analyze the DDC mRNA expression in HNSCC patients. METHODS: 53 malignant tumors were resected from the larynx, pharynx, tongue, buccal mucosa, parotid glands, and nasal cavity, as well as from 34 adjacent non-cancerous tissues of HNSCC patients, and were homogenized. Total RNA was isolated and converted into first-strand cDNA. An ultrasensitive real-time PCR method based on the SYBR Green chemistry was used for DDC mRNA quantification in head and neck tissue specimens. Relative quantification was performed using the comparative Ct (2-ddCt) method. RESULTS: DDC mRNA levels were lower in squamous cell carcinomas (SCCs) of the larynx and tongue than in adjacent non-cancerous tissue specimens. Furthermore, low DDC mRNA expression was noticed in laryngeal and tongue tumors of advanced TNM stage or bigger size, compared to early-stage or smaller tumors, respectively. No statistically significant differences were observed between SCCs resected from pharynx, buccal mucosa, or nasal cavity, and their normal counterparts. CONCLUSION: This is the first study examining the DDC mRNA expression in HNSCC. According to our results, DDC mRNA expression may constitute a potential prognostic biomarker in tongue and/or larynx SCCs, which principally represent the overwhelming majority of HNSCC cases.

Nonsurgical management of silent sinus syndrome with hyaluronic acid gel.

A 44-year-old woman was referred with 3 mm of left enophthalmos and a deep superior sulcus. She was diagnosed 6 months earlier with silent sinus syndrome and had undergone a left middle-meatal antrostomy with improvement of her hypoglobus but not of her enophthalmos and superior sulcus deformity. Two milliliters of Restylane SubQ (Q-Med AB, Uppsala, Sweden) was injected as a single bolus in the intraconal and extraconal posterior orbit via a standard transcutaneous inferotemporal approach using a primed green 21-gauge needle. Immediate reduction of enophthalmos by approximately 2 mm and superior sulcus improvement were noted. This remained stable at 6 months. The symptoms and signs of silent sinus syndrome can be effectively corrected with this minimally invasive nonsurgical option. To our knowledge, the use of hyaluronic acid gel for the treatment of enophthalmos in silent sinus syndrome has not been previously reported.

Continuation of smoking after treatment of laryngeal cancer: an independent prognostic factor?.

AIM: The aim of the study is to examine if continuation of smoking after treatment is an independent factor affecting the prognosis of laryngeal cancer. MATERIALS AND METHODS: A total of 153 patients met the inclusion criteria for this prospective study, and they were followed up for 12-60 months. Smoking cessation/continuation rates were recorded and associated with disease recurrence and overall patient survival. RESULTS: The recurrence rate was 35.29%. Twenty-five percent of the patients continued smoking after treatment, 75% stopped. Of the patients who quit smoking, 28.69% died during the follow-up period, compared to 52.63% of those who continued (p = 0.0047). The respective recurrence rates were 28.7 and 55.26% (p = 0.0022). A stepwise multivariate Cox regression analysis eliminated potential confounders regarding the overall survival rate and confirmed that time between symptom onset and diagnosis, T and N stage and continuation of smoking after treatment are statistically significant factors. Among them, continuation of smoking was found to have the strongest correlation to the overall survival rate. CONCLUSION: Continuation of smoking after treatment of laryngeal cancer is an independent negative prognostic factor. From a clinical standpoint, all patients with known laryngeal cancer should be strongly encouraged to quit smoking.

Vestibular evoked myogenic potentials in patients with BPPV.

BACKGROUND: The probable cause of Benign Paroxysmal Positional Vertigo (BPPV) is a degeneration of the oto lithic organs (utricle and saccule). The aim of the study is to find possible alterations in Vestibular Evoked Myogenic Potentials (VEMP) recordings in BPPV patients, because the saccule is part of the VEMP pathway. MATERIAL/METHODS: 27 BPPV patients (24 unilateral and 3 bilateral) aged 20 to 70 years and 30 healthy age matched controls. BPPV was diagnosed by the upbeating geotropic nystagmus found in the supine position with the head overextended towards one side. The subjects were investigated with pure tone audiometry, bi-thermal caloric test with electronystagmographic (ENG) recording, and VEMP recording. RESULTS: P1 latency and N1 latency did not present any statistical difference between control ears and affected ears of the BPPV population. The percentage of abnormal VEMP in the BPPV population was statistically higher than in the control ears (p < 0.005). No significant relationship could be shown between the occurrence of Canal Paresis and abnormal VEMP. No relationship was found between the side (right or left ear) where BPPV appeared clinically and the side where abnormal VEMP was registered. CONCLUSIONS: BPPV is a clinical entity associated with increased occurrence of abnormal VEMP recordings, possibly due to degeneration of the saccular macula, which is part of the neural VEMP pathway.

Supracricoid hemilaryngopharyngectomy for selected pyriform sinus carcinoma patients--a retrospective chart review.

BACKGROUND: The aim of this study is to assess the functional and oncologic results of supracricoid hemilaryngopharyngectomy and report our experience in the technique, local control and overall survival rates. MATERIALS AND METHODS: 18 selected patients with pyriform sinus cancer treated by supracricoid hemilaryngopharyngectomy in a University Hospital setting. Retrospective chart review was used to assess functional and oncologic results of the procedure. RESULTS: The actuarial 5 year survival rate in our study was 55.56% and the actuarial neck recurrence rate was 16.67%. All patients were successfully decannulated. Aspiration pneumonia was the most common postoperative complication (22.23%) and was treated mostly conservatively. One patient required a temporary gastrostomy but no patient needed total laryngectomy in the postoperative period. CONCLUSION: Supracricoid hemilaryngopharyngectomy in experienced hands is a reliable technique for selected patients with pyriform sinus cancer.

Resection of giant ethmoid osteoma with orbital and skull base extension followed by duraplasty.

BACKGROUND: Osteomas of ethmoid sinus are rare, especially when they involve anterior skull base and orbit, and lead to ophthalmologic and neurological symptoms. CASE PRESENTATION: The present case describes a giant ethmoid osteoma. Patient symptoms and signs were exophthalmos and proptosis of the left eye, with progressive visual acuity impairment and visual fields defects. CT/MRI scanning demonstrated a huge osseous lesion of the left ethmoid sinus (6.5 cm x 5 cm x 2.2 cm), extending laterally in to the orbit and cranially up to the anterior skull base. Bilateral extensive polyposis was also found. Endoscopic and external techniques were combined to remove the lesion. Bilateral endoscopic polypectomy, anterior and posterior ethmoidectomy and middle meatus antrostomy were performed. Finally, the remaining part of the tumor was reached and dissected from the surrounding tissue via a minimally invasive Lynch incision around the left middle canthus. During surgery, CSF rhinorrhea was observed and leakage was grafted with fascia lata and coated with bio-glu. Postoperatively, symptoms disappeared. Eighteen months after surgery, the patient is still free of symptoms. CONCLUSION: Before management of ethmoid osteomas with intraorbital and skull base extension, a thorough neurological, ophthalmological and imaging evaluation is required, in order to define the bounders of the tumor, carefully survey the severity of symptoms and signs, and precisely plan the optimal treatment. The endoscopic procedure can constitute an important part of surgery undertaken for giant ethmoidal osteomas. In addition, surgeons always have to take into account a possible CSF leak and they have to be prepared to resolve it.

Juvenile nasopharyngeal angiofibroma stages I and II: a comparative study of surgical approaches.

OBJECTIVE: The aim of this study was to compare different surgical options used for removal of stages I and II juvenile nasopharyngeal angiofibromas (JNAs). Treatment morbidity was evaluated through blood loss, surgery duration, postoperative hospitalization and outcome. Moreover, an effort was made to explore the role and limits of endoscopic surgery. MATERIALS AND METHODS: 20 male patients (mean age 14.9 years) were treated for JNA using three different surgical approaches in the Department of Otolaryngology of the University of Athens between May 1998 and January 2007. 9 patients were managed using endoscopic approach, while 5 were treated through midfacial degloving. A transpalatal approach was performed in remaining 6 patients. Preoperative angiography with embolization was performed in all 9 patients who underwent endoscopic removal and in 3 patients treated by midfacial degloving technique. RESULTS: Findings demonstrated that endoscopic approach, assisted by preoperative embolization, lead to less intraoperative blood loss, shorter duration of surgical procedure, shorter length of hospital stay and no complications, compared with the conventional techniques. CONCLUSIONS: Our data suggest that with proper patient selection, endoscopic resection of stages I and II JNA, when it is performed after embolization of the feeding vessels, is remarkably bloodless and precise and may be preferable to traditional open approaches.

Acoustic stapedial reflex in normal adults: biological behavior and determination of threshold levels.

OBJECTIVE: To record acoustic reflex thresholds for admittance (Y) and its components, susceptance (B) and conductance (G) in healthy subjects and to establish normative data. STUDY DESIGN: Acoustic reflex threshold was determined in 100 healthy persons, divided into 5 age groups of 20 persons each (20-29, 30-39, 40-49, 50-59 and 60-69 years old). Detection was performed for Y, B, and G at 226-, 678- and 1,000-Hz probe tones, using 500-, 1,000-, 2,000- and 4,000-Hz stimuli. RESULTS: Statistically significant differences in acoustic reflex thresholds were revealed between different components, stimuli and tones. The lowest threshold level was recorded for Y at 226 Hz using stimulus 1,000 Hz. Four percent missing values were observed at 226 Hz. Subjects belonging to different age groups had different reflex thresholds. CONCLUSION: Inclusion of acoustic reflex detection for B and G offers more detailed information about the middle ear status compared with Y alone, and appears to be sensitive in detecting subtle diseases.

Association between squamous cell carcinoma of the head and neck and serum folate and homocysteine.

UNLABELLED: The aim of this study was to evaluate the serum levels of folate and homocysteine in patients with squamous cell carcinoma of the head and neck (SCCHN) and to correlate them with the clinical stage. An attempt was made to associate the results with the effects of smoking. MATERIALS AND METHODS: Serum levels of folate and homocysteine were measured in 150 patients with histologically-proven SCCHN before any treatment and in 150 healthy volunteers (77 smokers and 73 nonsmokers). RESULTS: The study indicated a positive correlation between hyperhomocysteinemia and hypofolatemia and the presence of SCCHN. Folate deficiency and high levels of homocysteine were noted in a large number of healthy smokers. CONCLUSION: If metabolic disorders of the methionine cycle are confirmed as risk factors for SCCHN, folate as a dietary supplement might play a role in chemoprevention and the posttreatment care of SCCHN patients.

Clinical effectiveness of tumor markers in squamous cell carcinoma of the larynx.

BACKGROUND: The aim of this study was to examine and compare the value of the serum tumor markers CYFRA 21-1, TPA-M, SCCA and CEA in squamous cell carcinoma (SCC) of the larynx. Moreover, their possible clinical applications were studied. MATERIALS AND METHODS: The serum levels of CYFRA 21-1, TPA-M, SCCA and CEA were measured in 79 patients with histologically-proven squamous cell carcinoma (SCC) of the larynx before and after treatment and in 77 healthy volunteers. The association of the results with the clinicopathological characteristics was investigated. RESULTS: The study showed that none of the markers revealed a significant sensitivity; TPA-M indicated a positive correlation with the grade of differentiation. CEA indicated a positive correlation only with distant metastasis. CONCLUSION: Among the four markers, only TPA-M may play a role in monitoring the success of therapy and follow-up in patients with SCC of the larynx.

Inverted papilloma of the sphenoid sinus: presentation of three cases.

INTRODUCTION: Inverted papilloma deriving originally from the sphenoid sinus is extremely rare. Three such cases are presented aiming to show the particularities and diagnostic clues of this clinical entity. CASE REPORTS: All three patients were male, and presented mainly with nasal symptoms. In all three cases imaging shows the sphenoid sinus was completely occupied by the tumor. Additionally the ethmoids and nasal cavities have been invaded in contiguity to the site of the initial appearance of the tumor. In particular one of the cases was of even more interest as a malignant transformation of the inverted papilloma of the sphenoid sinus was histologically observed. Endoscopic sinus surgery was the therapeutic approach chosen for all three cases, with very good results. Follow up 30 months after the operation has not revealed any signs of recurrence of the disease. CONCLUSION: Inverted papilloma of the sphenoid sinuses--and even more malignant transformation--although quite rare, may present occasionally. The ENT surgeon should be aware of this condition. Preoperative imaging with CT scan and MRI and the experience of the surgeon will dictate the appropriate surgical approach.

Corneal topography with upper eyelid platinum chain implantation using the pretarsal fixation technique.

PURPOSE: To determine the effect of upper eyelid platinum chain implantation, with the pretarsal fixation technique, on corneal astigmatism. METHODS: This is a prospective, cohort study. Fifteen eyes of 15 patients underwent upper eyelid platinum chain implantation, with the pretarsal fixation technique, for facial nerve palsy. Information recorded included patient demographics, etiology for facial palsy, weight of the implant, time from onset of paresis to upper eyelid platinum chain implantation, associated surgical procedures, and preoperative and postoperative corneal topography measurements. RESULTS: Of the 15 patients studied, 10 were male and five were female. The mean age was 55.9 ± 13.8 years (range, 33-87 years). The most common etiology for facial palsy was acoustic neuroma. The weight of the implant ranged from 0.6 to 1.6gr (median 1.2gr). The time from onset of paresis to upper eyelid platinum chain implantation varied from 1 week to 3 months (median 1 month). Four patients had an associated procedure to correct the effect of paralytic ectropion. There was no statistically significant difference in with the rule astigmatism before and after platinum chain implantation. CONCLUSIONS: Upper eyelid platinum chain implantation, with the pretarsal fixation technique, does not appear to cause significant change in corneal astigmatism. This is contrary to data for pretarsal gold weight implantation, which does induce significant with the rule astigmatism.

Oral candidiasis in head and neck cancer patients receiving radiotherapy with amifostine cytoprotection.

This controlled study assessed the incidence of oral candidiasis, a xerostomia-related complication, in head and neck cancer patients receiving radiotherapy, with amifostine cytoprotection. Thirty-eight patients received 500 mg amifostine i.v., prior to each radiotherapy fraction, while 16 patients received radiotherapy alone. Oral candidiasis was diagnosed according to the criteria described before. Subjective xerostomia scales were completed by all patients. Mucositis was evaluated using the RTOG criteria. Oral candidiasis was diagnosed in 11/38 amifostine patients and in 9/16 controls (P = 0.07). Severe xerostomia was reported by 4/38 amifostine patients and by 7/16 controls. Oral candidiasis was reduced with amifostine cytoprotection. Oral candidiasis is suggested as an objective, early, though indirect, endpoint for amifostine's radioprotective effect on salivary glands.

14-3-3 gene expression exerts isoform-dependent functions in sinonasal pathophysiology.

The expression profiles of 14-3-3ß and θ isoforms, known to exert both oncogenic and antiapoptotic effects, were assessed in different entities of nasal pathophysiology. Flow cytometry and immunohistochemistry were used on paraffin-embedded sections of 51 inverted papillomas (IP), 26 nasal polyps (NP), 9 polyps with IP (NPIP) and 10 specimens of normal epithelium (NE). 14-3-3ß expression was significantly upregulated in IP as compared with both NP (p=0.015) and NE (p=0.002). 14-3-3ß was also increased in NPIP as compared with NE (p=0.008). 14-3-3ß cytoplasmic staining was more pronounced in basal cells of the respiratory epithelium although serous glands and the vascular system were often positive as well. High 14-3-3ß immunopositivity in IP patients concurred with increased proliferative activity shown by PCNA immunostaining (p=0.04). Expression of 14-3-3θ was also found increased in IP and NPIP patients, compared to NP (p=0.005, p=0.002 respectively) and NE (p=0.004 and p=0.001 respectively). 14-3-3θ cytoplasmic immunopositivity was detected in columnar epithelium, particularly in basal and subluminal cells, whereas no immunoreactivity was observed in NP and NE. Our results demonstrate differential expression of 14-3-3ß and θ isoforms in sinonasal pathophysiology, supporting their implication, respectively, in the proliferative and inflammatory process engaged in the formation of IP.

Diagnostic and prognostic significance of human kallikrein 11 (KLK11) mRNA expression levels in patients with laryngeal cancer.

OBJECTIVES: Human kallikrein 11 gene (KLK11) encodes a secreted serine protease. In view of its diagnostic and prognostic strength in many malignancies, we investigated the mRNA expression levels of KLK11 in laryngeal tissues in order to unveil its clinical usefulness in laryngeal cancer. DESIGN AND METHODS: KLK11 expression was quantified in 163 tissue samples from 105 laryngeal cancer patients with the development of a highly sensitive real-time PCR methodology, using SYBR Green® chemistry. RESULTS: KLK11 expression in laryngeal cancer specimens of primary or recurrent nature was significantly inferior compared with their non-malignant counterparts (P<0.001 and P=0.026, respectively), a finding of immense diagnostic value as illustrated in the ROC curve analyses (P<0.001). Survival analysis showed that patients harboring KLK11-positive tumors had a significantly decreased risk of death (HR=0.26, P=0.042). CONCLUSIONS: Our data recommend KLK11 mRNA expression as a novel and independent biomarker in laryngeal cancer for diagnostic and prognostic purposes.

l-DOPA Decarboxylase (DDC) Expression Status as a Novel Molecular Tumor Marker for Diagnostic and Prognostic Purposes in Laryngeal Cancer.

l-DOPA decarboxylase (DDC) plays an essential role in the enzymatic synthesis of dopamine and alterations in its gene expression have been reported in several malignancies. Our objective was to analyze DDC messenger RNA (mRNA) and protein expression in laryngeal tissues and to evaluate the clinical implication of this molecule in laryngeal cancer. In this study, total RNA was isolated from 157 tissue samples surgically removed from 100 laryngeal cancer patients. A highly sensitive real-time polymerase chain reaction methodology based on SYBR Green I fluorescent dye was developed for the quantification of DDC mRNA levels. In addition, Western blot analysis was performed for the detection of DDC protein. DDC mRNA expression was revealed to be significantly downregulated in primary laryngeal cancer samples compared with their nonmalignant counterparts (P = .001). A significant negative association was also disclosed between DDC mRNA levels and TNM staging (P = .034). Univariate analysis showed that patients bearing DDC-positive tumors had a significantly decreased risk of death (hazard ratio = 0.23, P = .012) and local recurrence (hazard ratio = 0.32, P =.006), whereas DDC expression retained its favorable prognostic significance in the multivariate analysis. Kaplan-Meier curves further demonstrated that DDC-positive patients experienced longer overall and disease-free survival periods (P = .006 and P = .004, respectively). Moreover, DDC protein was detected in both neoplastic and noncancerous tissues. Therefore, our results suggest that DDC expression status could qualify as a promising biomarker for the future clinical management of laryngeal cancer patients.

Apoptotic and proliferative status in HPV (+) and HPV (-) inverted papilloma patients. Correlation with local recurrence and clinicopathological variables.

Inverted papilloma (IP) is a rare sinonasal benign lesion characterized by aggressive biological behavior. Our aim was to evaluate the expression of various proliferation and apoptotic markers and the presence of HPV genotypes in paraffin sections gathered from surgically treated IP patients. Immunohistochemistry for PCNA, bax, cytochrome c and caspase-8 and flow cytometry for the detection of apoptosis, necrosis and ki67 expression were performed. The identification of various HPV subtypes was achieved by nested PCR amplification. Nasal polyps (NP) and specimens from normal nasal epithelium (NE) were used as controls. PCNA was more frequently expressed in IP compared to NE (p=0.04) and caspase-8 and bax staining were less frequently observed in IP compared to NP (p=0.004 and p=0.01 respectively) and NE (p=0.003 and p=0.01, respectively). IP and NP presented significantly higher Ki67 flow cytometry values compared to NE (p<0.001 and p=0.02 respectively). Cytochrome c was more frequently expressed in IP specimens with more prominent inflammation (p=0.02). A low HPV DNA detection rate was observed. Neither HPV status nor any of the apoptotic or proliferative markers studied was associated with the patients' clinicopathological characteristics. Increased Ki67 appeared to correlate with disease recurrence (p=0.01). Increased PCNA and Ki67 and decreased bax and caspase-8 expression indicate that cell proliferation is increased while apoptosis is inhibited in IP, explaining its biological behavior.