Current-generation high-throughput sequencing: deepening insights into mammalian transcriptomes.

Recent papers have described the first application of high-throughput sequencing (HTS) technologies to the characterization of transcriptomes. These studies emphasize the tremendous power of this new technology, in terms of both profiling coverage and quantitative accuracy. Initial discoveries include the detection of substantial new transcript complexity, the elucidation of binding maps and regulatory properties of RNA-binding proteins, and new insights into the links between different steps in pre-mRNA processing. We review these findings, focusing on results from profiling mammalian transcriptomes. The strengths and limitations of HTS relative to microarray profiling are discussed. We also consider how future advances in HTS technology are likely to transform our understanding of integrated cellular networks operating at the RNA level.

Retinoblastoma RB94 enhances radiation treatment of head and neck squamous cell carcinoma.

PURPOSE: To assess whether adenovirus-mediated retinoblastoma 94 (Ad-RB94) transgene expression enhances efficacy of radiation therapy (XRT) of human head and neck squamous cell carcinoma (HNSCC). EXPERIMENTAL DESIGN: The HNSCC cell lines (JHU006 and JHU012) were treated in vitro and in a nude mouse xenograft model with Ad-RB94, Ad-DL312 control vector, or untreated as mock control. Cell viability and tumor growth were evaluated and combined RB94/XRT antitumor activity was analyzed by measuring DNA double-strand breaks, apoptosis-associated early DNA fragmentation, and levels of RB-regulated cell cycle progression E2F1 transcription factor. RESULTS: Ad-RB94/XRT resulted in significant HNSCC cell growth inhibition compared with XRT alone or Ad-RB94 alone in vitro and caused significant tumor regression compared with XRT alone and Ad-DL312/XRT in JHU006 and with XRT alone, Ad-DL312/XRT and Ad-RB94 alone in JHU012 in vivo. Neutral comet analysis revealed that DNA damage was significantly elevated in cells treated with Ad-RB94 alone and Ad-RB94/XRT. Tumors treated with Ad-RB94 alone showed a striking increase in early apoptosis DNA fragmentation, and DNA fragmentation was further enhanced with XRT. In addition, levels of E2F1 were up-regulated by Ad-RB94/XRT combination, whereas Ad-RB94 alone did not affect E2F1 levels and XRT alone led to down-regulation of E2F1. CONCLUSIONS: A potent antitumor effect has been observed after Ad-RB94/XRT combination treatment in HNSCC xenograft tumors. Enhanced tumor regression correlated with increased apoptosis. Ad-RB94 treatment enhances the efficacy of XRT through tumor cell sensitization by arresting the cells at the radiation-sensitive G(2)-M cell cycle and via E2F1 up-regulation.

Airway obstruction: a rare complication of benign vocal fold polyps.

OBJECTIVES: Benign laryngeal polyps usually present with hoarseness and dysphonia. There have been a few reported cases, however, of polyps that caused airway obstruction. Herein we present our series of obstructing laryngeal polyps. METHODS: A retrospective review was performed of all patients with benign laryngeal lesions treated by the senior author (A.M.S.S.) between 1997 and 2006. Patients who presented with airway obstruction were identified. Detailed information was recorded on the demographics, presenting signs and symptoms, and surgical procedures. Preoperative and postoperative laryngoscopies were reviewed. RESULTS: Ten patients were identified. There were 7 women and 3 men. The mean age was 49 years (range, 34 to 64 years). All had a history of vocal abuse and smoking, with a mean of 35.2 pack-years (range, 7.5 to 112.5 pack-years). All underwent microlaryngoscopy with excision of the lesions. Jet ventilation or a small endotracheal tube was used to secure the airway. The mean follow-up of 9 patients was 71.2 days (range, 25 to 208 days); 1 patient was lost to followup. An excellent airway and improved voice was achieved in all. Forty-four percent (4 of 9) had persistent dysphonia and a decreased mucosal wave after the operation. CONCLUSIONS: Benign laryngeal polyps may present with airway obstruction and thus should be included in the differential diagnosis of stridor. Endoscopic treatment can result in an excellent airway, but dysphonia may persist in some cases.

Congenital anomalies of the larynx.

Congenital laryngeal anomalies are relatively rare. However, they may present with life-threatening respiratory problems in the newborn period. Associated problems with phonation and swallowing may prevent a baby from thriving. Stridor is the most common presenting symptom of congenital laryngeal abnormalities. Often, it is associated with dysphagia, aspiration, and failure to thrive. Endoscopy is essential for evaluation and diagnosis in most cases. The differential diagnosis includes laryngeal cysts, atresia and stenosis, vocal fold immobility, and subglottic hemangiomas. In this article, the authors discuss in detail the evaluation and treatment for each condition.

Thyroidectomy for Graves' disease: a case-control study.

OBJECTIVES: The most common treatment for Graves' disease in the United States is radioactive iodine. Surgery is performed rarely. The surgery for Graves' disease is usually considered technically difficult. Our goal was to assess the differences in outcomes in patients with Graves' disease who underwent thyroidectomy and in patients without Graves' disease who underwent the same procedures. METHODS: A retrospective chart review was performed of patients who underwent surgery for Graves' disease between 1997 and 2005. A control group of age-matched and thyroid size-matched patients who underwent thyroidectomy for a diagnosis other than Graves' disease was identified. The groups were statistically compared with respect to length of hospital stay, operative time, and estimated blood loss. Comparison with the published literature was also performed. RESULTS: Eleven patients underwent thyroidectomy for a diagnosis of Graves' disease. The operative time, estimated blood loss, and length of hospitalization did not differ significantly from those of the control patients. Three of the 4 complications that occurred, however, were in the 3 patients with persistent hyperthyroidism despite medical therapy at the time of surgery. CONCLUSIONS: Thyroidectomy may be performed relatively safely for selected euthyroid patients with Graves' disease. In those with persistent hyperthyroidism at surgery, there were more complications.

Feasibility of endoscopic treatment of middle ear myoclonus: a cadaveric study.

Stapedius and tensor tympani tenotomy is a relatively simple surgical procedure commonly performed to control pulsatile tinnitus due to middle ear myoclonus and for several other indications. We designed a cadaveric study to assess the feasibility of an entirely endoscopic approach to stapedius and tensor tympani tenotomy. We performed this endoscopic ear surgery in 10 cadaveric temporal bones and summarized our experience. Endoscopic stapedius and tensor tympani section is a new, minimally invasive treatment option for middle ear myoclonus that should be considered as the first line surgical approach in patients who fail medical therapy. The use of an endoscopic approach allows for easier access and vastly superior visualization of the relevant anatomy, which in turn allows the surgeon to minimize tissue dissection. The entire operation, including raising the tympanomeatal flap and tendon section, can be safely completed under visualization with a rigid endoscope.

The role of aerodigestive tract endoscopy in penetrating neck trauma.

OBJECTIVE/HYPOTHESIS: To determine the role of aerodigestive tract endoscopy in the management of penetrating neck trauma. STUDY DESIGN: Retrospective case series. METHODS: A search of the hospital's trauma database was performed for patients who presented with penetrating neck trauma between July 1989 and June 2008. The mechanism and site of injury, airway status and manipulation, physical findings, diagnostic and therapeutic steps taken, and outcomes were all recorded. RESULTS: One hundred sixty-three patients were identified. There were 144 males and 19 females. The mean age was 28 years (range, 13 to 65 years). There were 105 gunshot wounds, 9 shotgun injuries, and 48 stab wounds. Seventy-three patients (45%) underwent emergent neck exploration, of which 15 had upper aerodigestive tract injuries; intraoperative endoscopy was performed on 13 and was used to guide the repair. Ninety patients (55%) did not meet the criteria for emergent neck exploration. Endoscopy in this group was performed in symptomatic patients, which revealed two cases of unilateral true vocal fold motion impairment, one mucosal laceration of the right mainstem bronchus, one questionable area of injury at the apical segment of the right upper lobe bronchus, and one mucosal laceration of the proximal esophagus. CONCLUSIONS: Aerodigestive tract endoscopy is critical in assessing and guiding surgical repair of injuries noted on surgical exploration. In stable patients with a low clinical suspicion of aerodigestive tract injury, it was of low yield. We propose a new algorithm for assessing aerodigestive tract injuries that includes multidetector computed tomography, in which only symptomatic patients who fail to meet the criteria for emergent neck exploration undergo endoscopy. LEVEL OF EVIDENCE: 4.

Balloon laryngoplasty for pediatric laryngeal stenosis: case series and systematic review.

OBJECTIVES/HYPOTHESIS: The aim of this study was to systematically review available literature on the outcomes of children treated with balloon laryngoplasty (BLP) as a primary or adjuvant treatment for subglottic or laryngeal stenosis, as well as briefly report on a new series of 60 children treated at the Medical University of South Carolina from 2007 to 2013. STUDY DESIGN: Review of published case series and retrospective chart review. METHODS: A literature search was performed in PubMed and MEDLINE to identify trials that reported clinical outcomes of BLP in human patients under the age of 18 with subglottic or laryngeal stenosis. Single case reports and series studying the dilation of tracheal or bronchial stenosis alone were excluded. Hospital billing codes were used to identify appropriate patients for retrospective chart review. A successful outcome for chart review was determined to be decannulation of previous tracheostomy or avoidance of open laryngotracheoplasty or tracheostomy. RESULTS: Seven studies published between 1991 and 2012 met inclusion criteria and reported outcomes with success defined through improvement of symptoms, decrease in Myer-Cotton level of stenosis, decannulation, or avoidance of reconstructive procedures. Including 60 children from our institution, 202 patients between 1 day and 22 years of age (average 35 months) underwent 457 dilations, with an average of 2.26 dilations per patient (2.25 in our population). The overall success rate was 64% (77% in our population). No complications were reported with subglottic or laryngeal dilations. CONCLUSIONS: BLP is a highly effective, low-risk alternative or adjunct to traditional reconstructive procedures in children with subglottic or laryngeal stenosis. LEVEL OF EVIDENCE: 4.

Cerebellopontine angle ganglionic hamartoma: case report.

Cerebellopontine angle (CPA) ganglionic hamartomas are rare. We present a case of a 49-year-old woman who presented with left-sided hearing loss of more than 4 years' duration and vertigo since childhood. Magnetic resonance imaging revealed a mass on the intracanalicular segment of cranial nerve VIII on the left side, consistent with acoustic schwannoma. She opted for observation of the tumor. The patient returned 21 months after the initial evaluation with new-onset left facial weakness. At this time, a working diagnosis of facial nerve neuroma was made. The lesion was excised, and pathology confirmed a diagnosis of left-sided CPA ganglionic hamartoma. Otolaryngologists should be familiar with this uncommon tumor and include it in the differential diagnosis of CPA lesions.

Impact of neck dissection on long-term feeding tube dependence in patients with head and neck cancer treated with primary radiation or chemoradiation.

BACKGROUND: The impact of posttreatment neck dissection on prolonged feeding tube dependence in patients with head and neck squamous cell cancer (HNSCC) treated with primary radiation or chemoradiation remains unknown. METHODS: We conducted a retrospective cohort study using propensity score adjustment to investigate the effect of neck dissection on prolonged feeding tube dependence. RESULTS: A review of 67 patients with node-positive HNSCC (T1-4N1-3), treated with primary radiation or chemoradiation, with no evidence of tumor recurrence and follow-up of at least 24 months, was performed. Following adjustment for covariates, the relative risk (RR) of feeding tube dependence at 18 months was significantly increased in patients treated with posttreatment neck dissection (RR 4.74, 95% confidence interval [CI] 2.07-10.89). At 24 months, the relative risk of feeding tube dependence in the patients having undergone neck dissection increased further (RR 7.66, 95% CI 2.07-10.89). Of patients with feeding tubes 2 years after completing treatment, 75% remained feeding tube dependent. CONCLUSION: Neck dissection may contribute to chronic oropharyngeal dysphagia in HNSCC patients treated with primary radiation or chemoradiation.

Postradiotherapy neck dissection for head and neck squamous cell carcinoma: pattern of pathologic residual carcinoma and prognosis.

BACKGROUND: For patients with head and neck cancer who were treated using primary radiotherapeutic approaches, the pattern of pathologic residual carcinoma in the neck dissection specimen and its effect on clinical outcome remains unknown. METHODS: Medical records of 65 patients who underwent 71 neck dissections a median 7 weeks after radiotherapy were reviewed. Median follow-up was 33 months. RESULTS: Residual cancer, identified in 28 patients (43%), diminished locoregional control (p = .018), recurrence-free (p = .018), and overall survival (p = .02). Thirteen patients (20%) had 2 or more pathologically involved lymph nodes. Nine (13%) involved level V. Four (6%) had pathologic involvement of nodal levels not clinically involved by cancer before treatment. In N2-3 patients with positive pathologic specimens, the presence of these factors diminished recurrence-free survival (p = .01). The outcome of patients with pathologic carcinoma but without such ominous factors approached those with negative pathology. CONCLUSIONS: For patients with residual carcinoma in the neck following radiation, the pattern of residual disease is an effective predictor of recurrence.

Comparative analysis of tropism between canarypox (ALVAC) and vaccinia viruses reveals a more restricted and preferential tropism of ALVAC for human cells of the monocytic lineage.

The poxviruses including canarypox (ALVAC) and vaccinia viruses are promising vaccine vectors in humans, but little is known about their biology in human cells. Using recombinant enhanced green fluorescence protein (EGFP)-expressing ALVAC and vaccinia viruses, we have focused here on a side-by-side comparison of ALVAC and vaccinia virus tropism for cells from human peripheral blood and bone marrow. Both ALVAC and vaccinia viruses showed a strong bias towards monocyte infection. ALVAC minimally infected CD19+ B cells and was unable to infect ex vivo NK cells and T lymphocytes, whereas vaccinia virus could infect B lymphocytes and NK cell populations. Vaccinia virus was also able to infect T lymphocytes at low, but detectable levels that could be enhanced upon their activation. The observed preferential infection of ALVAC or vaccinia virus to monocytes was the result of preferential binding to this population, rather than lineage-specific differences in the expression of viral genes. Moreover, the level of CD14 expression on monocytes correlated with their preference to be infected with ALVAC or vaccinia virus. Both ALVAC and vaccinia viruses could infect immature monocyte derived dendritic cells (MDDCs), but only ALVAC infection induced their subsequent maturation. Vaccinia virus, however, showed greater tropism for mature MDDCs compared to ALVAC. Infection in human bone marrow cultures showed that ALVAC infection was restricted to a myelomonocytoid cell-specific CD33(+) cell population, while vaccinia virus showed a strong, but not exclusive, preference for these cells. These findings have implications in terms of choosing optimal pox virus derived vectors as vaccines in terms of reducing clinical reactogenicity and inducing dendritic cell (DC) maturation.