Targeted gene transfer into head and neck squamous cell carcinoma by nanosecond pulsed laser-induced stress waves.

The search for new strategies to improve patient outcome and organ preservation is of great clinical interest in head and neck squamous cell carcinoma (HNSCC) treatment, and gene therapy is expected to play a promising role. In this study, we demonstrated the value of laser-induced stress waves (LISWs) as a novel method for nonviral gene transfer for gene therapy in HNSCC. The in vitro and in vivo transfection efficiency as well as in vitro cytotoxicity for HNSCC was investigated. Green fluorescent protein (GFP) expression and cell viability were analyzed in vitro after administration of a GFP-expressing plasmid or cationically modified GFP-expressing plasmid with LISW application. Luciferase expression in xenograft tumors was also quantitatively analyzed in vivo. The GFP gene was successfully transfected into HNSCC cells in vitro by LISW application. The cationically modified plasmid demonstrated enhanced transfection efficiency. LISWs are not associated with adverse effects after application to cells in vitro. The reporter genes were also successfully transfected into HNSCC tumors in vivo by LISW application. This technique is site specific, safe, and easily applicable for practical purposes. LISW gene therapy with therapeutic factors that inhibit tumor growth therefore has the potential as a future treatment for HNSCC.

Composite hyoid bone graft interposition for the treatment of laryngotracheal stenosis.

INTRODUCTION: Chronic laryngotracheal stenosis (LTS) remains a challenging problem for otolaryngologists. A composite hyoid bone interposition graft has the potential to be an ideal graft because the head and neck surgeon can obtain the graft in the same operative field with good vascular supply from the muscle pedicle. METHODS: A composite hyoid interposition graft was used to provide structural support for the reconstructed lumen of the larynx or trachea in 2 cases of LTS. RESULTS: Two patients underwent successful decannulation with acceptable laryngeal function over a long-term observation period. CONCLUSION: This technique allows vascularized stable graft survival with minimal donor site morbidity. Furthermore, it can be performed for thyroid, cricoid, and tracheal stenosis without fear of damage to the recurrent laryngeal nerves.

[Impact of 131I SPECT/CT on the management of differentiated thyroid carcinoma outpatients with radioablation].

OBJECTIVE: The purpose of this study was to comparatively assess the diagnostic accuracy between 131I SPECT/CT and 131I whole-body scintigraphy (WBS) in differentiated thyroid carcinoma outpatients following radioablation. METHODS: a retrospective cohort study was performed in eleven patients with differentiated thyroid carcinoma (fourteen studies), who underwent both 131I WBS and 131I SPECT/CT, 7-10 days after administration of 1,110 MBq of 131I for radioablation, following a total thyroidectomy. A head and neck surgeon and two nuclear medicine specialists first interpreted the WBS images, followed by SPECT/CT images. RESULTS: SPECT/CT led to accurate revisions of the first diagnoses in 13 of 24 cervical foci of 131I uptake on WBS. Out of the 5 distant lesions, which were diagnosed as metastases by WBS alone, 5 (100%) were found out to be benign lesions by SPECT/CT. Thus, WBS was prone to be false-positive both in detecting cervical node metastases (13/28, 46.4%) and distant metastases (5/17, 29.4%). Eventually, SPECT/CT after 131I radioablation altered postoperative patients' management in 42.8% of patients with thyroid carcinoma by upstaging or down-staging their disease. CONCLUSION: SPECT/CT after 131I radioablation for differentiated thyroid cancer patients determines more accurately cervical lymph node metastasis, thyroid remnants and distant metastasis than does WBS alone. Both WBS and SPECT/CT after 131I radioablation are highly recommended for appropriate management of differentiated thyroid cancer patients who have undergone a total thyroidectomy.

Characteristic Video Laryngeal Endoscopic "Pharyngeal Rotation" in Unilateral Pharyngeal Constrictor Muscle Paresis: A Case of Herpes Zoster Pharyngitis.

Herpes zoster pharyngitis (HZP) is a rare condition that should be considered as a differential diagnosis of acute dysphagia secondary to unilateral glossopharyngeal and/or vagal nerve palsy. Although early treatment is important to avoid adverse sequelae, serological diagnosis of varicella zoster virus (VZV) takes over a few days. Therefore, it is important to actively suspect VZV infection based on physical findings. Mucocutaneous lesions, curtain signs, and laryngeal palsy are well-known characteristic physical findings. In addition to these findings, the video laryngeal endoscopic finding that the pharyngeal constrictor muscles contract on only one side during swallowing, showing an appearance of "pharyngeal rotation", is one of the characteristic findings of glossopharyngeal/vagal nerve palsy and can support the diagnosis. We report the case of an 82-year-old Asian female who presented with acute dysphagia, sore throat, left ear pain, and fever that persisted for several days. Initial video laryngeal endoscopy revealed a markedly decreased pharyngeal reflex and significant salivary retention without mucosal vesicular lesions. Repeat videoendoscopic evaluation of swallowing revealed characteristic pharyngeal rotation, which was helpful in diagnosing unilateral pharyngeal constrictor muscle paresis, thus suggesting unilateral glossopharyngeal/vagal nerve palsy. An increase in serum antibody titers (IgG and IgM) against VZV was observed. Bilateral differences and rotation of the pharynx during pharyngeal contraction can be detected endoscopically in pharyngeal constrictor muscle paresis caused by glossopharyngeal/vagal nerve palsy and should be evaluated during video laryngeal endoscopy in patients with dysphagia.

Comparison of voice quality after laser cordectomy with that after radiotherapy or chemoradiotherapy for early glottic carcinoma.

PURPOSE OF THE STUDY: The present study aimed to compare the voice quality after each type of cordectomy with that after radiotherapy (RT) or chemoradiotherapy (CRT) for early glottic carcinoma. PROCEDURES: The GRBAS perceptive scale [consisting of 5 domains: grade (G), roughness (R), breathiness (B), asthenia (A), and strain (S)], aerodynamic tests and acoustic analyses, and the Voice Handicap Index questionnaire were evaluated for 58 laser cordectomy cases and 40 RT or CRT cases. Multiple comparison tests were conducted between each type of cordectomy and RT or CRT. RESULTS: No statistical difference was found between type I/II cordectomy and RT for T1 glottic carcinoma (T1RT), whereas T1RT showed a significantly better outcome than type III cordectomy (G score: p = 0.016, maximum phonation time: p < 0.01, mean flow rate: p < 0.01). Type IV cordectomy was equivalent to RT or CRT for T2 glottic carcinoma (T2RT/CRT), while types V and VI showed a worse voice quality than T2RT/CRT (G score: p = 0.038 to type VI, B score: p = 0.025 to type V and p = 0.032 to type VI, A score: p = 0.017 to type VI). CONCLUSIONS: Voice quality after laser cordectomy differs according to the type of cordectomy. Surgeons should inform patients about the expected voice quality after each treatment modality.

A Case of Pancreatic Metastasis From Small Cell Neuroendocrine Carcinoma of the Oropharynx.

Small cell neuroendocrine carcinoma (SNEC) rarely occurs in the head and neck and usually occurs in the lungs. We report the case of a 55-year-old Asian male with SNEC in the oropharynx and jaundice due to pancreatic metastasis, which was successfully palliated by amrubicin (AMR), radiotherapy, and an endoscopic biliary stent. Although pancreatic metastases are known to occur at the end stage of small cell lung cancer, there are limited data on the treatment protocols for pancreatic metastases from SNEC. The main complication of SNEC for pancreatic lesions is obstructive jaundice. Palliative radiotherapy and biliary drainage may have life-prolonging effects in patients with extrahepatic biliary obstruction. It may also be a worthwhile risk to use anticancer drugs, such as AMR that are metabolized in the liver, if the obstructive jaundice is caused by tumor growth.

Vocal function after transoral videolaryngoscopic surgery (TOVS) for hypopharyngeal and supraglottic cancer.

CONCLUSION: The resection of the medial and lateral pyriform sinus was associated with post-operative voice impairment after TOVS. Scar contracture around the cricoarytenoid joint lead to arytenoid fixation toward lateral position, and this wound healing process caused insufficient glottis closure. Although oncological and functional outcomes of TOVS was satisfactory, surgeons should mention the risk of post-operative voice impairment in pre-operative counseling. OBJECTIVES: Transoral surgery is a minimally invasive treatment option for hypopharyngeal and supraglottic cancer. Post-operative vocal function was satisfactory in most cases, but in some cases vocal cord was fixed and occasionally voice impairment persists. METHODS: Vocal function of 55 patients who underwent transoral videolaryngoscopic surgery (TOVS) for hypopharyngeal and supraglottic cancers was evaluated by the GRBAS perceptive scale, aerodynamic tests and acoustic analyses, and the Voice Handicap Index questionnaire. The risk factors for voice impairment were identified. RESULTS: Voice impairment (G score ≧2) was found in 16 cases (29.1%). Univariate analysis revealed that the resection of medial and lateral pyriform sinus (p = .0018) and neck dissection (p = .0421) were associated with post-operative voice impairment. Multivariate analysis revealed that the resection of medial and lateral pyriform sinus (p = .0021) was associated with post-operative voice impairment.

Risk factors for dysphagia after transoral videolaryngoscopic surgery for laryngeal and pharyngeal cancer.

BACKGROUND: Transoral surgery is gaining attention as a minimally invasive treatment option for laryngeal and pharyngeal cancer. Postoperative swallowing function is satisfactory in most cases, but occasionally dysphagia persists. METHODS: Swallowing function of 86 patients who underwent transoral videolaryngoscopic surgery (TOVS) for laryngeal and pharyngeal cancers was evaluated by the Functional Outcome Swallowing Scale (FOSS) and risk factors for dysphagia were identified. RESULTS: FOSS stage was 0 to 1 in 90%, 2 in 5.8%, 3 in 3.5%, and 4 in 1.2% of the patients. Only 1 patient had indication for percutaneous endoscopic gastrostomy (PEG). Univariate analysis revealed that resection of the pyriform sinus (p = .0280) and arytenoid (p = .0139), pulmonary dysfunction (p = .0353), and large mucosal defect (p = .0223) were associated with postoperative dysphagia. CONCLUSION: Although the rate of PEG dependency is low in transoral surgery, surgeons should mention the risk of postoperative dysphagia when counseling patients preoperatively.

Serum midkine as a biomarker for malignancy, prognosis, and chemosensitivity in head and neck squamous cell carcinoma.

Improved therapies for individuals with head and neck squamous cell carcinoma (HNSCC) may be developed by identification of appropriate biomarkers. The aim of this study was to evaluate the usefulness of serum midkine measurement as a biomarker for HNSCC. Pretreatment serum midkine concentrations were measured in 103 patients with HNSCC and 116 control individuals by enzyme-linked immunosorbent assay. Midkine expression in tumor tissues from 33 patients with HNSCC who underwent definitive surgical resection without preoperative treatment was examined by immunohistochemistry. The cut-off serum midkine concentrations for predicting the presence of head and neck malignancy and chemosensitivity to induction chemotherapy, as determined using receiver operating characteristic curves, were 482 and 626 pg/mL, respectively. Spearman bivariate correlations showed positive correlations between serum midkine levels and immunohistochemistry staining score (r = 0.612, P < 0.001). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of serum midkine concentration for detection of HNSCC were 57.3, 85.3, 77.6, 69.2, and 72.1%, respectively. However, for predicting the response to induction chemotherapy, the values were 84.6, 60.9, 71.0, 77.8, and 73.5%, respectively. Serum midkine concentration was identified as an independent prognostic factor by multivariate analysis, using Cox's proportional hazards model (P = 0.027). Overexpression of serum midkine yielded a relative risk of death of 3.77, with 95% confidence limits ranging from 1.15 to 17.0. Serum midkine levels in patients with HNSCC were associated with malignancy, chemosensitivity, and prognosis. Serum midkine may be a useful, minimally invasive biomarker for early detection, therapeutic decision-making, and predicting prognosis.

Tacrolimus prevents laryngotracheal stenosis in an acute-injury rat model.

OBJECTIVES/HYPOTHESIS: Acquired laryngotracheal stenosis is a challenging problem for otolaryngologists. Several studies suggest tacrolimus may inhibit post-transplant airway stenosis that occurs with coronary drug-eluting stents. The objective of the present study was to determine whether tacrolimus modulates wound healing of the airway mucosa and prevents laryngotracheal stenosis in an acute injury animal model. STUDY DESIGN: Basic science. METHODS: The laryngotracheal mucosa of rats was scraped with a nylon brush through the tracheostoma. Tacrolimus (0.2 mg/kg or 1.0 mg/kg) was systemically administered intramuscularly for 5 days. Nine days after scraping, the pathological changes and the degree of stenosis were assessed by hematoxylin and eosin staining or by immunohistochemical staining for nuclear factor of activated T cell and interleukin 2. RESULTS: Lumen stenosis resulted from hyperplasia of the airway epithelium and a thickened submucosal layer with extensive fibrosis, angiogenesis, and collagen deposition. There was a significant preventive effect on airway stenosis at the tracheal and cricoid levels in the low-dose (0.2 mg/kg) tacrolimus-treated animals, compared to the untreated animals (P < .05). This effect was insignificant with treatment by high-dose tacrolimus (1.0 mg/kg). Immunohistochemistry showed that, after tacrolimus treatment, the expressions of nuclear factor of activated T cell and interleukin 2 were downregulated in submucosal fibroblasts, neovascular cells, and glandular cells. CONCLUSIONS: This study suggests that low-dose systemic tacrolimus has a preventive effect on laryngotracheal stenosis by inhibiting the activation of immune cells in the injured airway mucosa via the calcineurin/nuclear factor of activated T cell/interleukin 2 pathway. LEVEL OF EVIDENCE: NA.

Gene therapy of c-myc suppressor FUSE-binding protein-interacting repressor by Sendai virus delivery prevents tracheal stenosis.

Acquired tracheal stenosis remains a challenging problem for otolaryngologists. The objective of this study was to determine whether the Sendai virus (SeV)-mediated c-myc suppressor, a far upstream element (FUSE)-binding protein (FBP)-interacting repressor (FIR), modulates wound healing of the airway mucosa, and whether it prevents tracheal stenosis in an animal model of induced mucosal injury. A fusion gene-deleted, non-transmissible SeV vector encoding FIR (FIR-SeV/ΔF) was prepared. Rats with scraped airway mucosae were administered FIR-SeV/ΔF through the tracheostoma. The pathological changes in the airway mucosa and in the tracheal lumen were assessed five days after scraping. Untreated animals showed hyperplasia of the airway epithelium and a thickened submucosal layer with extensive fibrosis, angiogenesis, and collagen deposition causing lumen stenosis. By contrast, the administration of FIR-SeV/ΔF decreased the degree of tracheal stenosis (P < 0.05) and improved the survival rate (P < 0.05). Immunohistochemical staining showed that c-Myc expression was downregulated in the tracheal basal cells of the FIR-SeV/ΔF-treated animals, suggesting that c-myc was suppressed by FIR-SeV/ΔF in the regenerating airway epithelium of the injured tracheal mucosa. The airway-targeted gene therapy of the c-myc suppressor FIR, using a recombinant SeV vector, prevented tracheal stenosis in a rat model of airway mucosal injury.

Novel Indocyanine Green-Phytate Colloid Technique for Sentinel Node Detection in Head and Neck: Mouse Study.

OBJECTIVE: Sentinel node navigation surgery using real-time, near-infrared imaging with indocyanine green is becoming popular by allowing head and neck surgeons to avoid unnecessary neck dissection. The major drawback of this method is its quick migration through the lymphatics, limiting the diagnostic time window and undesirable detection of downstream nodes. We resolved this problem by mixing indocyanine green (ICG) with phytate colloid to retard its migration and demonstrated its feasibility in a nude mouse study. STUDY DESIGN: Experimental prospective animal study. SETTINGS: Animal laboratory. SUBJECTS AND METHODS: Indocyanine green at 3 concentrations was tested to determine the optimal concentration for sentinel lymph node detection in a mouse model. Effect of indocyanine green with phytate colloid mixture solutions was also analyzed. Indocyanine green or mixture solution at different mixing ratios were injected into the tongue of nude mice and near-infrared fluorescence images were captured sequentially for up to 48 hours. The brightness of fluorescence in the sentinel lymph node and lymph nodes further downstream were assessed. RESULTS: Indocyanine green concentration >50 µg/mL did not improve sentinel lymph node detection. The addition of phytate colloid to indocyanine green extended the period when sentinel lymph node was detectable. Second echelon lymph nodes were not imaged in mice injected with the mixture, while these were visualized in mice injected with indocyanine green alone. CONCLUSION: This novel technique of ICG-phytate colloid mixture allows prolonged diagnostic time window, prevention of downstream subsequent nodes detection, and improved accuracy for the detection of true sentinel lymph nodes.

"Reverse" bypass using a naturally formed "bonnet" superficial temporal artery in symptomatic common carotid artery occlusion: a case report.

We herein describe a patient with symptomatic common carotid artery occlusion who underwent a reverse superficial temporal artery (STA)-middle cerebral artery (MCA) single bypass using a naturally formed "bonnet" STA. The surgical procedure was performed without difficulty, and no further neurological deterioration was observed after surgery. In practice, this case highlights that the reverse STA-MCA bypass can be achieved safely and less invasively using a naturally formed "bonnet" STA.

Superparamagnetic iron oxide-enhanced interstitial magnetic resonance lymphography to detect a sentinel lymph node in tongue cancer patients.

CONCLUSION: This is the first report on human sentinel node (SN) detection by interstitial magnetic resonance (MR) lymphography with superparamagnetic iron oxide (SPIO) in tongue cancer patients who also underwent lymphoscintigraphy. Our results indicate that further studies are warranted, as this novel method may replace current scintigraphic techniques. OBJECTIVES: To examine the feasibility of interstitial MR lymphography using SPIO for SN detection in the head and neck region. METHODS: MR images were acquired sequentially at 10 min, 30 min, and 24 h after submucosally injecting 0.1 ml SPIO (ferucarbotran) around the tumor in three patients with tongue cancer without cervical lymph node metastasis (clinical T2N0M0). RESULTS: The SNs were clearly visualized in the 10 min interstitial MR lymphography images and were completely concordant with those visualized by (99m)Tc-radiocolloid lymphoscintigraphy and a gamma probe in all cases. Iron incorporation into the SNs was confirmed by pathological examination.

Sendai virus transgene in a novel gene therapy for laryngotracheal disease.

OBJECTIVES/HYPOTHESIS: Vocal cord scar formation and laryngotracheal stenosis (LTS) are challenging problems for otolaryngologists. Sendai virus (SeV) vectors have been shown to transduce airway epithelium efficiently, and are thus ideal for modulating airway wound-healing therapy. To assess the potential utility of SeV gene therapy for laryngotracheal diseases, we established a novel LTS model and examined the transduction efficiency of SeV vectors in normal and LTS model tissue. STUDY DESIGN: Basic science. METHODS: Fusion (F) gene-deleted, nontransmissible SeV vectors were used. First, the route dependency and transduction efficiency of SeV vectors for normal mucosa in the larynx were examined. Next, the novel LTS rat model was established and evaluated. Finally, the transduction efficiency of SeV vectors in injured mucosa of the LTS model was evaluated. RESULTS: Simple spray delivery of the SeV vector resulted in significant and persistent expression of the reporter gene in normal laryngotracheal epithelium. Transgenic SeV-mediated expression was maximal at 3 days, decreased over time, but remained detectable for 14 days after administration. No serious side effects were observed in the larynx or trachea. The model achieved an average of 60% tracheal stenosis in the cross-sectional area. Efficient SeV-mediated transgene expression was observed in the injured mucosa at the levels of the trachea, cricoid cartilage, and vocal cord. CONCLUSION: A novel animal model for LTS was established. We successfully demonstrated SeV-mediated transgene expression in normal tissue and in the injured mucosa of the LTS model. SeV might be a promising strategy for gene therapy in laryngotracheal diseases.

Comparison of animal studies between interstitial magnetic resonance lymphography and radiocolloid SPECT/CT lymphoscintigraphy in the head and neck region.

OBJECTIVE: To comparatively assess two techniques, radiocolloid SPECT/CT lymphoscintigraphy and interstitial MR lymphography using SPIO and gadoxetate disodium, in animal models. MATERIALS AND METHODS: We used twenty one 8-week-old male nude mice of strain BALB/c Slc-nu/nu, weighing 23-27 g. The 4.7-T MRI equipment was used to detect the SNs. T2*WI of gradient-echo sequences was acquired sequentially up to 24 h after administering SPIO, ferucarbotran. T1WI was acquired sequentially up to 80 min after administering gadoxetate disodium. (99m)Tc-phytate SPECT/CT lymphoscintigraphy was taken at 30 min after the injection to detect the SNs using animal-dedicated whole-body SPECT/CT hybrid scanner. The injection was submucosally performed in the right tongue margin of each mouse. Reading performances concerning SN visualization and its quality on interstitial MR lymphogram and SPECT/CT lymphoscintigram were performed by 3 radiologists. RESULTS: The SN intensities were 0.43 for the right, 0.61 for the left at 30 min after ferucarbotran injection, with gradual decrease in intensity, and 1.43 for the right, 1.33 for the left at 10 min after gadoxetate disodium injection with a fast decrease in intensity. The base value of 1.0 was at pre-examination. The mean numbers of lymph nodes visualized were 4.00 nodes for on SPECT/CT lymphoscintigram and 2.0 for interstitial MR lymphogram. There was a statistically significant difference in the mean scores between SPECT/CT lymphoscintigraphy and interstitial MR lymphography (two factor mixed design with repeated measures on one factor: p < 0.0002). CONCLUSIONS: In our comparative study using mice, the results of radiocolloid SPECT/CT lymphoscintigraphy were superior to those of interstitial MR lymphography, while both SPIO and gadoxetate disodium have a potential of being employed for sentinel node navigation surgery by interstitial MR lymphography in the head and neck region.

Relationship between globus sensation and esophageal clearance.

CONCLUSIONS: Videofluoroscopy (VF) in the prone position can diagnose a range of esophageal pathologies in all age groups. These correlate with globus sensation in younger and middle-aged patients in prone and supine positions. Abnormal esophageal clearance appears to be associated with globus sensation in young and middle-aged persons, but not in the elderly. OBJECTIVES: To assess the correlation between globus sensation and abnormal esophageal clearance by VF in relation to body position and to investigate age-related abnormal esophageal clearance, as well as to confirm the utility of VF in diagnosing globus sensation in patients. SUBJECTS AND METHODS: This was a prospective study. In 72 patients with globus sensation and 33 controls, esophageal clearance was assessed by VF in the upright, supine, and prone positions, and transfer of barium through the esophagus was classified as normal, esophageal retention, esophageal reflux, or pharyngeal reflux. The detection rate of abnormal esophageal clearance was compared among all subjects by age. RESULTS: In the upright position, VF detected no significant differences in the detection rate of abnormal esophageal clearance and reflux between the patients with globus sensation and controls; however, the detection rate of abnormal esophageal clearance was significantly higher in the prone than in the upright and supine positions (p<0.001), and in all positions was significantly higher among globus sensation patients (p<0.05). In the supine and prone positions, there were significantly higher detection rates of abnormal esophageal clearance in patients than in controls in the young group (p<0.05). Additionally in the prone position, there was a significantly higher detection rate of abnormal esophageal clearance and reflux in patients than in controls in the middle-aged group (p<0.05). In all positions, no significant differences were observed in the detection rate of abnormal esophageal clearance and reflux in the patients and controls in the elderly group.