bluuebunny/crossref_metadata_2025 huggingface dataset:
Connexins Within the Rat Larynx
Authors: Van Deusen Mark , Lyon Michael J
Published: August 2008

Problem

To determine the various types of connexins and their localization within the rat larynx.

Methods

Eight larynges from 3–4 month old Fisher‐344 rats were used. Four were immediately dissected to obtain the epiglottis, vocal folds, supraglottic and subglottic mucosa. RNA was extracted and used as a template for production of cDNA, which was used with primers for Cx26, Cx30, Cx32, Cx37, Cx40, and Cx43 and RT‐PCR performed. Four others were serially sectioned and the sections processed for immunohistochemistry.

Results

RT‐PCR revealed a high level of Cx43, Cx32, and Cx30 within epiglottis and Cx43 in the vocal folds. Immunohistochemical staining confirmed these results.

Conclusion

The rat epiglottis is rich in Cx43, Cx32, and Cx30 while the vocal folds contain Cx43. Their localizations suggest involvement in secretion for protective purposes and they may play a key role in laryngeal pathologies.

Significance

Cxs in the larynx may be disrupted by decreased pH, alcohol ingestion, and smoke inhalation. These three factors contribute significantly to the pathology that patients commonly present to the otolaryngologist. Potential pharmacologic treatments are on the horizon and further characterization of the laryngeal connexins needs to be carried out before such treatments can be utilized.
COX II Expression in Laryngeal Lesions
Authors: Brys Agata K. , Haben C Michael
Published: August 2008

Problem

Over expression of Cyclooxygenase II (COX II) is noted in a variety of inflammatory and neoplastic conditions, including laryngeal cancer. Although smoking is a known risk factor for laryngeal carcinoma, non‐smokers are occasionally diagnosed. Possible etiologic factors include up regulation of COX II expression. This study compares COX II expression in benign, dysplastic and cancerous laryngeal tissue specimens in smokers and non‐smokers.

Methods

Pathology specimens from consecutive patients who underwent excision of a laryngeal lesion for unrelated reasons from 2003 to 2006 were collected. Patients with severe laryngopharyngeal reflux were excluded. Patients’ smoking history was collected. Immunocytochemical studies were used to detect COX II expression. A blinded pathologist graded the intensity of COX II expression based on a visual analog scale. Statistical analysis was used to determine differences.

Results

49 samples from 22 smokers and 18 non‐smokers were tested. COX II expression was significantly larger in cancerous lesions of smokers vs non‐smokers and in dysplastic vs benign lesions in non‐smokers. A non‐statistically significant trend of consistent and progressive increase in COX II expression with increasing dysplasia in smokers was noted.

Conclusion

The increase in COX II expression between benign and dysplastic lesions in non‐smokers may be due to a loss of regulation of cell metabolism. This effect may be HPV mediated and further studies comparing HPV positivity with COX II expression are underway. The significantly higher COX II expression in cancerous lesions of smokers vs non‐smokers be due to the persistent and chronic mucosal irritation from smoking.

Significance

This translational research may affect treatment and prevention of laryngeal carcinoma in non‐smokers. A correlation between increased COX II expression and pathologic diagnosis would support the role for the use of COX II inhibitors in the prevention of progression of laryngeal lesions.

Support

Research grant from the Department of Otolaryngology, University of Rochester, New York.
GM‐CSF‐Mediated Amplification of Airway Fibrosis
Authors: Sethna Anita , Mattox Douglas E , Guidot David M , Mitchell Patrick
Published: August 2008

Problem

1. To determine if GM‐CSF (granulocyte macrophage‐colony stimulating factor) protects against fibrosis using an experimental animal model of tracheal injury. 2. To determine if GM‐CSF prevents cytokine‐induced expression of fibrotic markers in human bronchial epithelial cells in vitro.

Methods

Tracheae from GM‐CSF (20ng) or vehicle (PBS) treated Sprague‐Dawley donor rats were heterotopically transplanted into Fischer 344 recipients. Recipients were treated daily with either GM‐CSF or vehicle for seven days. Allografts were harvested on days 14 or 21 post‐transplantation and analyzed for luminal collagen deposition and the presence of intact epithelium. In parallel, human bronchial epithelial cells (Beas2B) were treated with pro‐fibrotic cytokines (TGFbeta1, IL‐13) with or without GM‐CSF. Treated cells were harvested and processed for RT‐PCR or immunocytochemical analyses for expression of fibrotic markers (alpha smooth muscle actin, fibroblast‐specific protein 1).

Results

Tracheal allografts treated with GM‐CSF demonstrated a significant increase in luminal collagen deposition compared to vehicle‐treated allografts. GM‐CSF‐ treatment was associated with destruction of airway epithelium, contributing to amplification of fibrosis. Beas2B cells treated with the same concentration of GM‐CSF demonstrated increased expression of markers of tissue remodeling and fibrosis as well as decreased epithelial markers.

Conclusion

We hypothesized that GM‐CSF‐mediated protection of the tracheal epithelium may prevent fibrosis of the airway. We show here that GM‐CSF increases luminal fibrosis and decreases epithelial viability in vivo. In addition, treatment of Beas2B cells with GM‐CSF in vitro increases expression of pro‐fibrotic markers. These data emphasize the critical role of the epithelium in preventing luminal fibrosis after tracheal injury.

Significance

Various mediators have been considered in the etiology of tracheal stenosis. Previous studies in our lab have demonstrated the presence of GM‐CSF receptors in tracheal epithelium. Our hypothesis was that protection of this epithelium could prevent airway fibrosis. The current study demonstrates that airway fibrosis correlates with a loss in epithelium induced by GM‐CSF.
Model for Examining Recovery of Phonation after Nerve Damage
Authors: Bhama Prabhat K. , Hillel Allen D , Merati Albert L , Perkel David
Published: August 2008

Problem

Recurrent laryngeal nerve injury remains a dominant clinical issue in laryngology. To date, no animal model of laryngeal reinnervation has offered an outcome measure that can reflect the degree of recovery based on vocal function. We present an avian model system for studying recovery of learned vocalizations following nerve injury.

Methods

Digital recordings of bird song were made from 11 adult male zebra finches; 9 underwent bilateral crushing of the nerve supplying the vocal organ, and two birds underwent sham surgery. Song from all birds was then recorded regularly and analyzed based on temporal and spectral characteristics using computer software. Indices were calculated to indicate the degree of similarity between pre‐operative and post‐operative song.

Results

Nerve crush caused audible differences in song quality and significant drops (p<0.05) in measured spectral and, to a lesser degree, temporal indices. Spectral indices recovered significantly (mean=43.0%; SD=40.7; p<0.02), and there was an insignificant trend towards recovery of temporal index (mean=28.0%; SD=41.4; p=0.0771). In 5/9(56%) birds, there was a greater than 50% recovery of spectral indices within a four week period. Two birds exhibited substantially less recovery of spectral indices, and two had a persistent decline in spectral indices. Recovery of temporal index was highly variable as well, ranging from persistent further declines of 45.1% to recovery of 87.0%. Neither sham bird exhibited significant (p>0.05) differences in song following nerve crush.

Conclusion

The songbird model system allows functional analysis of learned vocalization following surgical damage to vocal nerves.

Significance

The zebra finch provides an animal model for the study of phonatory function following neurolaryngeal injury.

Support

This project was supported by NIH grant T32DC000018.
RNAi Supress VEGF, TERT, and Bcl‐xl in HEp‐2 Cells
Authors: Tao Ze‐Zhang , Yan Wang
Published: August 2008

Problem

To develop an RNA‐interference (RNAi) approach involving the hitting of multiple targets by a recombinant plasmid and evaluate its antitumor effect on laryngeal squamous carcinoma in vitro and in vivo.

Methods

A plasmid containing 3 different short hairpin RNA (shRNA) segments termed pEGFP‐shVEGF‐shTERT‐shBcl‐xl was constructed. Plasmids containing single shRNA against each target (VEFG, TERT, BCL‐xl alone) individually were also constructed as control. Cells were treated with these plasmids. The expression of targeted genes as well as apoptosis of tumor cells were evaluated after treatment with multiple shRNA vectors or control vectors. The mRNA and protein expression were determined by RT‐PCR and western blotting. Cell viability was examined using the MTT assay. Apoptotic morphological alterations were observed by Hoechst staining and electron microscopy. The in vivo antitumor effect was characterized in a nude mice model of laryngeal squamous carcinoma.

Results

We demonstrated that a recombinant plasmid containing multiple shRNAs could effectively and simultaneously inhibit VEGF, TERT and Bcl‐xl mRNA and protein expression in the HEp‐2 cells; the plasmid containing the 3 different shRNAs exhibited a potent antitumor effect on LSCC both in vitro and in vivo, and could much more effectively induced cell apoptosis than each single shRNA. We also demonstrated that the simultaneous blockage of these 3 genes have a better inhibitory effect on human HEp‐2 cells than the blockage of each single shRNA.

Conclusion

Our study demonstrates that the application of vector‐based RNAi technology that involves hitting multiple targets will be a promising therapeutic modality in the gene therapy of human laryngeal cancers.

Significance

Our study provides experimental evidence for the clinical application of this technology in the future.

Support

This work was supported by the grants from the National Natural Science Foundation of China (No. 30471873, No.30672313 and No.30740012).
Atypical Pharyngitis In Suppurative Otitis Media Cases
Authors: Abou‐Bieh Alaa A , Salama Mona F
Published: August 2008

Problem

Unexplained persistent or recurrent bacterial pharyngitis in some patients who suffer from infected middle ear cleft.

Methods

Bacteriological swabs were obtained from both the ears and the pharynx of 37 cases with chronic otorrhea and perforation, whom complain of recurrent or persistent sore‐throat. Then isolation and identification of the micro‐organisms were done. This included examination of direct Gram stained films and cultures. Isolated Gram negative bacilli were subjected to further identification by the biochemical reactions and antibiotyping. Identical isolates from the same patient (ear and pharyngeal swabs) were subjected to further identification by genotyping using the pulsed field gel electrophoresis technique.

Results

6 cases (16%) showed identity in phenotypes and genotypes for ear and pharyngeal samples from the same patient. All pharyngeal isolates were Gram negative organisms. 4 of them were Pseudomonas aeruginosa, 1 was Proteus sp., and 1 was Escherichia coli. All of these 3 species are not known to be among the primary organisms which may cause pharyngitis.

Conclusion

Bacterial pharyngitis in patients with chronically infected middle ear cleft may be attributed to the same organism invaded the middle ear mucosa. Also this study highlights some organisms as a pharyngeal invaders although they are not among the previously documented causatives of bacterial pharyngitis. But the study do not confirm the method of spread of these organisms and whether this was directly via the eustachian tube.

Significance

The study correlates the causative organism of the middle ear infection and that infected the pharyngeal mucosa utilizing the advanced bacteriological identification and genotyping techniques.
Cochlear Soft Tissue Imaging Using In‐Line Phase Contrast
Authors: Fishman Andrew J , Christoph Rau , Lixin Fan , Richter Claus‐Peter
Published: August 2008

Problem

Most contemporary imaging methods require invasive specimen preparation and/or are time consuming, or lack sufficient spatial resolution.

Methods

Experiments were made at the Advanced Photon Source (APS), Argonne National Laboratory. The APS is a synchrotron radiation source of the third generation, for which the particular characteristic is the highly coherent X‐ray radiation. X‐rays are generated with an undulator, inserted in a straight section of the storage ring. Images taken with hard X‐rays and the grating interferometer were compared with images from the same specimen acquired with light microscopy.

Results

The results show that imaging of cochlear soft tissue structures is possible with hard X‐rays and a grating interferometer.

Conclusion

Imaging of cochlear soft tissue structures is possible with hard X‐rays and a grating interferometer.

Significance

It is important to examine cochlear morphology from many points of view, including comparative anatomy, cochlear developmental changes, malformation caused by genetic defects, changes related to diseases, sensory physiology, and cochlear modeling. Best imaging results were obtained from in situ experiments in the absence of tissue distortion. However, most contemporary imaging methods require invasive specimen preparation and/or are time consuming, or lack sufficient spatial resolution. A method that potentially overcomes these shortcomings is presented in this paper and it uses hard X‐rays and a grating interferometer. Furthermore, hard X‐rays would allow imaging cochlear soft tissue structures without opening the cochlea itself.

Support

The UNICAT facility at the Advanced Photon Source is supported by the U.S. DOE under Award No. DEFG02‐91ER45439, through the Frederick Seitz Materials Research Laboratory at the University of Illinois at Urbana‐Champaign, the ORNL (U.S. DOE contract DE‐AC.
Customized Electric Suppression of Tinnitus in CI Subjects
Authors: Fine Esther L , Qing Tang , Rothholtz Vanessa S , Lu Thomas , Djalilian Hamid R , Zeng Fan‐Gang
Published: August 2008

Problem

To determine the degree of tinnitus suppression in cochlear implant patients by systematically varying stimulation parameters as a function of rate, level, and electrode.

Methods

Five cochlear implant subjects with ipsilateral or bilateral tinnitus were included. Implants were stimulated at a basal, middle, and apical electrode, at 100pps and 5000 pps, at perceived level quieter than the tinnitus and at the most comfortable loudness level. Patients reported stimulus and tinnitus perception on a ten‐point scale every 30 seconds during and after stimulus delivery.

Results

All subjects reported near or complete suppression of tinnitus under one or more test conditions. Complete accommodation to the stimulus occurred often, thus during suppression patients perceived very little sound or total silence. Residual inhibition lasted up to twenty minutes. Two of the subjects noted suppression when low rate stimulation was delivered to an apical electrode. One patient responded only to high rate stimulation to an apical electrode. The remaining two patients reported suppression under multiple conditions. One subject had the most dramatic and consistent effects with low rate stimulation to a middle electrode; high rate stimulation caused rebound tinnitus of up to five minutes.

Conclusion

All patients experienced tinnitus suppression with electrical stimulation. Low rate stimulation overall was more effective in our subject group, and in two cases was the only effective suppressor.

Significance

Previous studies utilized only high rates of stimulation, and treatment failures occurred. Electrical stimulation is an effective, reproducible, and well‐tolerated treatment for tinnitus but stimulus customization is required on an individual basis.
Middle Ear Packing: Comparison Between Merogel and Gelfoam
Authors: Lipan Michael Julian , Ibraham Alava , Angeli Simon I , Van De Water Thomas R
Published: August 2008

Problem

Gelfoam has been used for decades in otologic surgery to support grafts and prostheses despite causing fibrosis and adhesions. More biocompatible packing materials could avoid these complications. This study compares Gelfoam with an injectable esterified hyaluronic acid, Merogel, as middle ear packing material after mucosal trauma.

Methods

A randomized, blinded, controlled study was performed in 17 juvenile guinea pigs. Middle ear surgery with mucosal trauma was performed and middle ears were packed with Merogel or Gelfoam; unpacked contralateral ears were used as controls. Auditory brainstem response (ABR) thresholds were measured in 4 frequencies pre‐operatively, and repeated at 1, 2, and 6 weeks postoperatively. Gross analysis measured inflammatory reaction in each group. Statistical analysis was performed using ANOVA followed by post‐hoc analysis for ABR thresholds and packing remaining at 6 weeks. Non‐prametric tests were used for presence of mucosal inflammation, effusion and tympanic membrane perforation.

Results

ABR threshold changes from baseline were minor and comparable between the Merogel group and the control group. Threshold change was higher in the Gelfoam group. This difference was seen in each frequency tested at each time interval (all p<0.05). Gross analysis showed 1) Mucosal inflammation higher in the Gelfoam group (p<0.05), 2) Effusions were higher in the Gelfoam group but the difference was not significant (p=0.07), 3) Tympanic membrane perforation was equally rare between groups (p>0.05), 4) Unabsorbed packing was higher in the Gelfoam group (p<0.05); little Merogel was detectable at time of sacrifice.

Conclusion

Middle ear healing after surgery occurred similarly in the control group and the Merogel group. In contrast, the Gelfoam group demonstrated greater perturbation of hearing and a greater inflammatory reaction.

Significance

These results support Merogel as an alternative to Gelfoam in middle ear packing after otologic surgery.

Support

Research grant from Medtronic ENT.
The Measurement of 2D Curvature of In‐Vivo Human Ear Canal
Authors: Yu Jen‐Fang , Chung‐chieh Fan , Chen Chin‐Kuo
Published: August 2008

Problem

To non‐invasively measure the 2D curvature of human ear canal and produce the earmold by the non‐invasive 3D ear impression.

Methods

The images of external ear were scanned by high‐resolution computed tomography (HRCT). The resolution for each slice was 512⋉512 pixels. The pixel size was 0.188⋉0.188mm and the slice thickness was 0.625mm. The boundary between tympanic membrane and external auditory meatus was enhanced by image processing. Additionally, 3D model of ear canal was reconstructed by 2D images. The length and angle of first and second bends of canal were measured based on the 3D model. 2D curvature of first and second bends of canal was then computed by sine and cosine laws.

Results

The volume of ear canal was 862.0 cubic mm. The angle and curvature of superior wall of first bend at axial view were 121.5 degrees and 0.0685; of inferior wall of first bend were 246 degrees and −0.1102; of superior wall of second bend were 227.8 degrees and −0.0332; of inferior wall of second bend were 143.1 degrees and 0.0130 respectively. 2D curvature of superior and inferior wall of first and second bends was diagrammed.

Conclusion

The 2D curvature of ear canal at first and second bends could be measured and produce the ear impression non‐invasively. The geometry of canal changed by tumors is a common syndrome in ear disease. Therefore, the geometry of ear canal can be tracked after the otoplasty.

Significance

The 3D geometry of canal can help physicians diagnose the syndrome of external canal before otoplasty. Moreover, the hearing aid earmold can be made by non‐invasive ear impression instead of invasive ear impression.
Volumetric Rendering of Human Inner Ear by MR Imaging
Authors: Yu Jen‐Fang , Chin Wei‐Chung , Wu Che‐Ming , Ng Shu‐Hang
Published: August 2008

Problem

To non‐invasively measure in‐vivo human inner ear by MRI and measure the geometry of vestibule by the reconstructed 3D model of inner ear for further diagnosis of large vestibular aqueduct syndrome (LVAS).

Methods

3‐T MR scanner, MAGNETOM Trio made by Siemens, was utilized. The TR/TE for MR imaging of 7 patients was 5.65/2.6 ms and the voxel size was 0.5 mm X 0.5 mm X 0.5 mm for single slice of 48 slices. The configuration of semicircular canals, vestibule and cochlea could be detected by threshold. The 3D geometry of inner ear was then computed based on the thickness of slice.

Results

The surface area and volume of semicircular canals for 7 normal ears were 217.85 square mm and 63.56 cubic mm; of vestibule were 105.88 square mm and 56.36 cubic mm; of cochlea were 171.84 square mm and 81.29 cubic mm respectively. The variation of volumes of vestibule and cochlea could be quantified non‐invasively. The correlation between the volume and the level of LVAS will be analyzed once the number of volunteer reaches a statistically significant level.

Conclusion

The variation for the geometry of vestibule could be measured non‐invasively. The grade of LVAS can be assessed by the obtained 3D model of semi‐circular canal, vestibule and cochlea.

Significance

According to the 3D model, the geometry of inner ear can be measured, and the syndrome can be revealed directly to help clinical diagnosis of LVAS more accurately.
Evaluation of the PEAK PlasmaBlade for ENT Surgery
Authors: Chang Kay W , Carlson Grace A , Eric Huang
Published: August 2008

Problem

Traditional electrosurgical instruments are known to have an associated zone of thermal injury to the residual tissue. The compromised tissue left behind in the body may compromise wound healing and also may cause post operative pain. The PEAK PlasmaBlade is a new tissue dissection tool that uses pulsed radiofrequency energy to generate a highly focused plasma field at the tip of the device. This creates an effective cutting edge with simultaneous hemostasis while the blade remains near body temperature. A comparative study of tissue cutting and hemostasis was conducted to evaluate the characteristics of this technology.

Methods

Porcine mucosal and lymphoid tissue was subjected to a series of surgical incisions using the PlasmaBlade, a scalpel, a traditional electrosurgery (bovie) blade style tip, and a Coblation EVac 70 wand. Both low and high settings were evaluated for PlasmaBlade, bovie, and Coblation. Blood loss following the cut was evaluated. Histology samples were harvested immediately after and evaluated for collateral tissue damage at the incision site. Thermal imaging of all the devices in an active cutting mode was also conducted.

Results

Histological evaluation of the tissue samples showed that the PlasmaBlade cuts produced minimal collateral damage compared to cuts made with the other electrosurgical instruments, which demonstrated greater thermal damage. Bleeding control (hemostasis) was equivalent for the Plasma‐Blade versus bovie and Coblation. Thermal imaging showed the PlasmaBlade operating in a range of 40 to 100 C which was similar to the Coblation operating temperatures. Traditional electrosurgery produced temperatures well in excess of 200 C.

Conclusion

The PEAK PlasmaBlade is a promising new instrument which provides atraumatic, scalpel–like, cutting and bovie‐like hemostasis, resulting in minimal bleeding and tissue injury.

Significance

There is great potential for this becoming an important tool for common ENT procedures such as tonsillectomy and adenoidectomy, specifically in reducing the pain and morbidity of recovery.
Fgf8 is Necessary for Normal Frontonasal Development
Authors: Wittkopf Maria , Goudy Steven L
Published: August 2008

Problem

Craniofacial development is a complex process. We explore the role of ectodermal Fgf8 during craniofacial development in mice. Fgf8 is a multi‐functional growth factor known to induce cell migration and differentiation. We hypothesize that Fgf8 is necessary for frontonasal development. To determine its role, we selectively removed Fgf8 signaling in cells expressing the AP2 gene.

Methods

We generated AP2 Cre Fgf8 f/f R26R mice at e13–18. The embryos are harvested and fixed. They are dehydrated or saturated in 30% sucrose, then embedded in paraffin or OCT compound, respectively, for sectioning. The paraffin sections are used for morphologic examination following H&E and PAS staining. The cryosections are used for X‐gal staining and immunofluorescence. Skeletal preparations are used to assess the craniofacial skeleton.

Results

AP2 Cre mice are examined identifying AP2 expression in the developing epithelium. Gross examination of the AP2 Cre Fgf8 f/f mice reveals severe mid‐face shortening and micrognathia. These mice are nonviable due to their severe facial abnormalities. Close inspection reveals prominent eyes that appear normally formed but lack eyelids. The skeletal preparation of the AP2 Cre Fgf8 f/f mouse demonstrates almost absent mandibular development and hypoplastic maxilla. This correlates with the hypoplastic, ill‐organized sinus cavities with lack of nasal septum development seen on histologic sections. Examination of the epithelium and oral mucosa of these mutant mice demonstrates a verrucous morphology with poor differentiation.

Conclusion

Fgf8 expression in the ectoderm appears to be necessary for frontonasal development in mice. Without epithelial Fgf8 expression, the craniofacial and epidermal development is severely affected. Future studies will further characterize the epithelial and mesenchymal phenotype of the AP2 Cre Fgf8 f/f mutant mice.

Significance

The Fibroblast Growth Factor (Fgf) family is involved in multiple steps of craniofacial formation. Further studies elucidating the specific roles of its members, including Fgf8, will help better understand and manage craniofacial malformations.
Growth Factors in Infantile Hemangiomas
Authors: Tan‐Geller Melin , Min‐Jung O Teresa , Zaida Alipio , Fink Louis , Yongming Ma , Waner Milton
Published: August 2008

Problem

Infantile hemangiomas have a well‐known distinct proliferative phase characterized by rapid proliferation of capillaries, followed by an involuting phase characterized by spontaneous and steady regression of the tumor. Histologically, proliferating hemangiomas are composed of masses of compact capillaries lined by plump endothelial cells with high mitotic rates. As the lesions involute, mitosis gradually decreases with increased apoptosis of endothelial cells, and vascular tissue is gradually replaced with fibrofatty tissue. Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) are among several markers that have been qualitatively found in proliferating and involuting phases of infantile hemangiomas, respectively. The purpose of this study is to quantitatively measure growth factors in proliferating and involuting hemangiomas.

Methods

A high‐throughput xMAP multiplex immunobead assay technology (Luminex Corp., Austin, TX) was used to test VEGF, FGF, endothelial growth factor (EGF), and granulocyte‐colony stimulating factor (G‐CSF) in 22 infantile hemangioma specimens, 8 plasma specimens from hemangioma patients, and normal skin. Results were then correlated to histologic findings at the time of surgical removal.

Results

When compared to plasma and normal skin as controls, infantile hemangiomas demonstrated increased amounts of VEGF and EGF in actively proliferating hemangiomas and increased amounts of FGF, in both proliferating and involuting hemangiomas. G‐CSF was insignificant in hemangioma specimens.

Conclusion

VEGF, EGF and FGF are upregulated in infantile hemangiomas and may represent important therapeutic targets.

Significance

This is the first study to quantitatively measure growth factors in infantile hemangiomas at different growth stages. Definitive evidence of increased growth factors in proliferating hemangiomas may lead to further consideration of angiogenic inhibitors as an alternative treatment modality to surgery, laser or steroids.

Support

This research study was supported by the Vascular Birthmarks Foundation.
The Leicester Olfactometer
Authors: Philpott Carl M , Goodenough Paul C , Allan Clark , Murty George E
Published: August 2008

Problem

The objective of this study was to develop a useful and cost‐effective olfactometer for routine clinical use. The apparatus was developed to provide a standardized threshold test for patients with olfactory disorders presenting in the ENT clinic.

Methods

A prospective study of olfactory thresholds in 48 healthy volunteers on 2 consecutive occasions, undergoing quantitative testing with an olfactometer. Further studies of 10 subjects performing 20 tests and 100 subjects performing a single test were also performed. An olfactometer was designed to deliver a semi‐automated threshold test for an odor. It was designed as a device containing 8 logarithmic dilutions of an odor along with a control valve operated by software from a laptop computer. Common potential variables for olfactory threshold testing were considered including peak inspiratory flow rate and smoking status. The odors used for the studies were phenethyl alcohol (PEA) and eucalyptol. Subjects were asked to perform 2 tests within 1 month of each other and the mean threshold score for each test was calculated to help derive a test‐retest score.

Results

Consistent olfactory thresholds for PEA were achieved with a mean concentration of 10–4. Test‐retest reliability scores for the olfactometer were rx = 0.78 (95% CI 0.67 to 0.89). Common variables thought to affect olfactory testing including peak inspiratory flow rate and smoking were not found to have a significant effect on the threshold scores.

Conclusion

The Leicester Olfactometer provides a simple and cost‐effective method of reliably assessing olfactory thresholds in the outpatient clinic.

Significance

Routine testing of olfaction can be provided with a sophisticated yet user friendly format that overcomes the expense of single use test kits and other commercially available kits.

Support

Departmental research funds used.
R406 – Treatment of Squamous Cell Carcinoma of the Hypopharynx
Authors: Chedid Helma Maria , Lehn Carlos Neutzling
Published: August 2008

Problem

Squamous cell carcinoma of the hypopharynx has been diagnosed at a late stage, including in developed countries, thus denoting worse prognosis. An association between surgery and radiotherapy is the standard treatment for advanced clinical stages. Our aim was to assess the clinical and treatment factors of tumors of the hypopharynx that were treated surgically and with postoperative radiotherapy.

Methods

This was a retrospective study on 174 patients, among whom two patients were in clinical stage (CS) I, four in CS II, 46 in CS III, and 122 in CS IV. Among these, 222 neck dissections were performed, of which 48 were bilateral. Statistic analysis: Kaplan‐Meier methods.

Results

The mean follow‐up was 31 months, and histological examinations showed that two patients were pT1, 15 pT2, 100 pT3 and 57 pT4. With regard to lymph nodes, 25 patients were pN0 and 149 patients were pN+. The disease‐free survival in CS III was 75%, versus 28% in CS IV (p=0.001). The disease‐free survival was 35% and the overall survival was 28%.

Conclusion

Surgical treatment for tumors of the hypopharynx occurred in advanced CS, with disease‐free survival of 35%.

Significance

The overall survival and disease free‐survival among patients with squamous cell carcinoma of the hypopharynx undergoing surgery.
R409 – Apigenin and Kaempferol Effects on Oral Cancer FADU Cells
Authors: Helton William B , Choi Eun‐Young , Gairola C Gary , Valentino Joseph , Hollie Swanson
Published: August 2008

Problem

Dietary flavonoids are being investigated as chemopreventative agents for many cancers. The objective of this study was to determine whether the dietary flavonoids apigenin and kaempferol inhibit growth of malignant oral keratinocytes.

Methods

FaDu cells were treated with increasing concentrations of apigenin and kaempferol. After 24 and 48 hours cell growth was determined using the WST‐1 assay. Three groups of nude mice (group 1‐kaempferol, group 2‐apigenin, group 3‐controls) were treated by gavage for one week prior to inoculation with FaDu cells (1 × 105 cells) Following inoculation, treatments were continued until sacrifice. Tumor volumes were calculated from three dimensional tumor measurements.

Results

In vitro cell growth decreased with increasing concentrations of apigenin and kaempferol (p<0.001). In vivo tumor volume was significantly higher than controls for the apigenin group (p<0.03) but was marginally higher in kaempferol group (p=0.09) with an average volume of 3024mm3, 2610 mm3, and 1858 mm3 respectively.

Conclusion

Apigenin and kaempferol inhibited tumor cell growth in culture. However, in vivo results show that these substances increased tumor burden. This is in contrast to previous in vivo and in vitro prostate cell line models showing apigenin inhibition of tumor growth. Further studies are needed to better evaluate the effect these dietary flavonoids exert on squamous cell carcinoma of the head and neck.

Significance

The role of these agents as chemopreventative agents for oral carcinoma is not supported by our data.

Support

University of Kentucky Department of Surgery Research Grant and NIH R‐01 grant.
R411 – FISH of Salivary Tumors to Enhance Diagnostic Accuracy
Authors: Kraai Tania L , Olson Garth T , Bocklage Therese J , John Hozier
Published: August 2008

Problem

Fine needle aspiration biopsy (FNAB) is used to triage patients with salivary tumors. Distinguishing benign from malignant is difficult on cytology features and immunohistochemistry alone, leading sometimes to an incorrect surgical resection. Therefore, we evaluated whether fluorescent in situ hybridization (FISH) could be performed routinely on salivary gland tumor FNAB's, potentially to improve diagnostic accuracy.

Methods

Pathology files from our institution were searched for patients with salivary gland tumors. We collected data on 74 tumors: 25 benign (18 pleomorphic adenoma and 7 Warthin's tumors) and 49 malignant (miscellaneous including adenoid cystic carcinoma, mucoepidermoid carcinoma and acinic cell carcinoma, among others). Clinical presentation, staging, and follow‐up information were obtained. Representative tumor blocks were disaggregated, converted to nuclear preps, and analyzed for aneuploidies of chromosomes 3, 7, 9, 11, and 19 using FISH. A novel technique creating a CMA (cytology microarray) enabled testing of multiple samples on one glass slide. Signal quantitation was performed using a Metasystems instrument. Shrunken centroid analysis was used to test for significant differences in the benign versus malignant group.

Results

Six tumors exhibited trisomies (8 percent); those few were exclusively limited to the malignant tumors. The tumor preps yielded adequate cellularity in 90% percent, and the CMA technique was time and cost efficient.

Conclusion

In a large number of salivary gland tumors, we found that chromosome copy number abnormalities do not occur with high enough frequency to enhance diagnostic accuracy. When trisomy occurs, it more likely is associated with malignancy.

Significance

Polyploidy seen on FISH analysis of the FNA of salivary tumors does not enhance diagnostic accuracy.
R412 – Role of KITENIN in Progression and Metastasis of SCC
Authors: Zarrella Mark , MD Joon Kyoo Lee , Lim Sang‐Chul
Published: August 2008

Problem

The expression of KAI1 (a metastatic suppressor gene) in cancer cells results in reduced cell motility and invasiveness. A cDNA clone of VANGL1, a member of the tetraspanin protein family that specifically interacts with the COOH‐terminal cytoplasm domain of KAI1, was isolated and renamed KITENIN (KAI1 COOH‐terminal interacting tetraspanin). The purpose of this study was to investigate the role of KITENIN on the progression and metastasis of transfected squamous cell carcinoma using in vivo and in vitro experiments.

Methods

Locally advanced squamous cell carcinoma tissues from five patients were obtained for investigation of KITENIN expression. Malignant tumors, normal adjacent mucosa tissues, metastatic lymph nodes, and non‐metastatic lymph nodes were collected. KITENIN or vector only (control) was transfected into SCC (squamous cell carcinoma) VII, a mouse squamous cell carcinoma cell line, using FuGENE 6. An in vitro assay (invasion, migration, and proliferation) for KITENIN and the vector‐transfected group was studied. The KITENIN or vector‐transfected SCC VII cells were injected subcutaneously into 12 C3H/HeJ syngeneic mice (6 mice for each group). The tumor size was measured daily for 4 weeks. During the fifth week after injection, the presence of metastasis in the lung and liver tissue was evaluated for each mouse with a tumor mass on the back; the tissues were assessed by gross and microscopic examination.

Results

KITENIN was highly expressed in tumors and metastatic lymph nodes from patients. KITENIN‐transfected cells showed significantly increased invasion, migration, and proliferation compared with the vector‐transfected cells. Tumor volume was more increased in the KITENIN‐transfected cells‐injected mice. Lung metastasis was found in the KITENIN‐group (3/6 mice), while no metastasis in the vector‐group.

Conclusion

KITENIN participates in the progression and metastasis of SCC.

Significance

An antisense KITENIN strategy may be a useful method to inhibit progression and metastasis in squamous cell carcinoma.

Support

This study was financially supported by Chonnam National University, 2006.
R414 – Nasopharyngeal Undifferentiated Carcinoma: Study of 46 Cases
Authors: Lehn Carlos Neutzling , Chedid Helma Maria , Porsani Alex Freitas
Published: August 2008

Problem

The nasopharyngeal carcinoma (NPC) has been a rare cancer in the world. The incidence is higher in the southern Asiatic, Groveland and Tunisia. The age range follows the bimodal distribution. It appears most frequently in males, with a ratio of 3:1. Carcinogenic factors such as tobacco and alcohol have a role of little importance in the carcinogenesis of nasopharyngeal undifferentiated carcinoma. The diagnosis has gotten in elderly phase, named noiseless phase of growing and is not common the initial presentation with metastatic lymph node in the neck. The aim of this study was the assessment of demographic, clinical and treatment factors of nasopharyngeal undifferentiated carcinoma in Department of Head and Neck Surgical and Otorrinolaryngology of Heliópolis Heliópolis‐Hosphel.

Methods

This was the retrospective study of 46 patients on January 1998 to august 2000. All patients with nasopharyngeal carcinoma had presented histopathological diagnostic of undifferentiated type. The treatment was the radiotherapy concomitant to chemotherapy to the clinical stage III and IV. Statistic analysis: Kaplan‐Meier methods.

Results

The symptom of the most importance was the lymph node of neck. The local recurrences were not retreat and three of four regional recurrences were treated with neck dissection. All patients that spread out distance metastasis had presented loco‐regional control.

Conclusion

The tobacco and alcohol were not factors related to disease presentation. The survival free of disease to three years was of 52%.

Significance

The incidence, clinical factors and treatments and survival among patients with nasopharyngeal undifferentiated carcinoma.
R415 – Recurrence of Nasopharyngeal Angiofibroma (NPF) Study
Authors: Vishnu Vardhan , Martha R.
Published: August 2008

Problem

Study of correlation of recurrence of NPF with its histopathologic appearance.

Methods

A retrospective and prospective study of 184 NPF cases identified, treated in Government Ear Nose Throat Hospital, and followed over a period of 26 years (1981–2006) and their histopathological slides are preserved, their recurrence rate is documented. The histopathology of NPF is varied ‐ composed of vascular and fibrous components and their proportion varied. In our study we will undertake the review of histopathology slides that are already preserved and would also like to study the new recurrent cases in the coming 6 months.

Results

All recurrences were observed with in one year of of treatment and the recurrence rate was 19.66%. Recurrence had no correlation with age of patient, duration of symptoms, or surgical approach but correlated with stage of tumor at presentation (p less than 0.05). Preoperative embolisation was done in 16.85% cases and did not show any statistical difference in recurrence rate between the embolised and non embolised.

Conclusion

The correlation between the histopathological appearance and the recurrence rate will help in choosing the right approach for surgery and also postoperative follow‐up to detect the recurrence at the earliest.

Significance

Histopathological detail can help in knowing the prognosis in terms of chances of recurrence and also guide for follow up at close intervals and advocate the appropriate treatment at the earliest.
R417 ‐ The Akt Pathway as a Biomarker in Carcinogenesis
Authors: Nathan Cherie Ann O , McEachern Matthew , Thornton Blakely Nelson , Rong Xiaohua , Clark Cheryl
Published: August 2008

Problem

The only definitive end point for studies of chemoprevention is the determination of the incidence of primary or secondary cancers which would require a multi‐year study with thousands of subjects and tremendous costs. The identification and validation of intermediate endpoints would be an important step in evaluating chemopreventive agents. We have previously noted that curcumin, an exciting nutraceutical with promising chemopreventive effects in HNSCC, inhibited the Akt pathway. Hence we wanted to determine if there was a difference in expression of pAkt in tumors and adjacent mucosa of HNSCC patients when compared to non cancer patients.

Methods

16 HNSCC and 16 non‐cancer (sleep apnea) patient samples were analyzed by western blot for expression of phosphorylated Akt (pAkt(Ser473)). The SCC samples include tumors and adjacent mucosa.

Results

Phosphorylated Akt was elevated 12‐fold in tumor samples compared to non‐cancer patients. No expression of pAkt was seen in mucosa of 12/16 non‐cancer patients while tumors and adjacent mucosa express pAkt in 15/16 HNSCC patients (p<0.05).

Conclusion

The pAkt/mTOR pathway may be sensitive as Akt/mTOR is activated in 99% of HNSCC and not uvulas suggesting pAkt is a good biomarker for a chemopreventive agent.

Significance

This preferential activation of the Akt pathway in HNSCC compared with the non‐cancer patients could be useful in the design of clinical trials with curcumin.

Support

This work was supported by the Feist‐Weiller Cancer Center and the American Society of Hematology.
R424 – Suppression of HNSCC with Liposomal Curcumin and Cisplatin
Authors: Suh Jeffrey D , Duarte Victor M , Salvado Amanda R , Veena Mysore Sadasivaiya , Wang Dorothy , Srivatsan Eri S , Wang Marilene B
Published: August 2008

Problem

To investigate whether curcumin, previously shown to inhibit growth of head and neck squamous cell carcinoma (HNSCC), can potentiate the anti‐tumor effects of cisplatin in vitro and in vivo.

Methods

HNSCC cells were treated with curcumin and cisplatin, individually and together, and assayed for growth using MTT, western blotting, and immunofluorescence. Five‐week old female athymic nude mice were utilized for in vivo experiments. HNSCC cells were injected to form xenograft tumors. Liposomal curcumin (50 ug/kg, maximum volume of 100ul) was administered intravenously three times per week for four weeks via tail vein injection. Mice were divided into three groups: no injection, liposomes alone, or liposomal cur‐cumin. At the end of the fourth week, a single dose of intraperitoneal cisplatin was given to all groups. At the end of five weeks, the mice were euthanized and tumors were dissected and weighed.

Results

Addition of curcumin or cisplatin alone or in combination resulted in cell death. Introduction of suboptimal levels of both agents in vitro demonstrated a synergistic suppression of HNSCC cell line growth compared to individual agents. Xenograft tumors in nude mice treated with combination curcumin and cisplatin showed superior tumor growth suppression compared to control mice and mice treated with liposome alone. Average tumor size for mice treated with combination therapy was 30% smaller than controls.

Conclusion

Curcumin potentiates the anti‐tumor activity of Cisplatin in vivo. Our data supports further investigation into the potential use of curcumin in combination with cisplatin in the treatment of HNSCC.

Significance

Current treatment modalities for HNSCC carry significant morbidity, which has led to the investigation of less toxic therapies. Previous studies by our group have demonstrated that liposome‐encapsulated curcumin can suppresses growth of HNSCC. In this study, we assess the potential of combining liposomal curcumin with cisplatin in the treatment of HNSCC.
R429 – Irradiation Changes MyHC Expression in Rat Larynx
Authors: Kim Jin‐Pyeong , Khalmuratova Roza , Jeon Sea Yuong , Park Jung Je , Hur Dong Gu , Kang Ki Mun
Published: August 2008

Problem

Laryngeal cancer treatment involves radiotherapy, laser treatment, and/or surgery. Radiotherapy does not impair voice quality as much as laser treatment or surgery but can induce muscle wasting, fibrosis, and dry mouth symptoms. To elucidate the molecular mechanisms involved in such changes, we investigated the effect of irradiation on myosin heavy chain (MyHC) expression in laryngeal muscles.

Methods

Rats were irradiated with one dose of 10, 15, 20, 25, 30, or 35 Gy and sacrificed 2 weeks later. Other rats were irradiated with 20 Gy and sacrificed 3 days or 2, 4, or 12 weeks later. The thyroarytenoid (TA), posterior cricoarytenoid (PCA), cricothyroid (CT), and digastric (DG) muscles were subjected to SYBR Green real time‐reverse transcriptase‐PCR.

Results

Radiation exceeding 20 Gy killed all rats within 7‐12 days. Two weeks after irradiation with 10, 15, or 20 Gy, MyHC type IIa expression had decreased in a dose‐dependent manner in the PCA and CT muscles and in a dose‐independent manner in the TA muscle. The expression of MyHC IIx and IIL also decreased but in a dose‐dependent manner in the TA muscle and a dose‐independent manner in the PCA and CT muscles. In the 20 Gy‐irradiated rats, MyHC IIx, IIL, and especially IIa expression declined over 12 weeks.

Conclusion

The laryngeal muscles responded differently to radiation but showed a time‐dependent and long‐lasting decrease in the expression of all MyHC isoforms. MyHC IIa expression in the PCA and CT muscles may be more sensitive to irradiation than the other MyHC isoforms.

Significance

All of these observations together support the notion that if the radiation‐induced changes in MyHC expression in the larynx can be ameliorated by an as‐yet undefined method, it may be possible to restore to normal the laryngeal muscles of larynx cancer patients who have been trated with radiation therapy, therepy presevering their voice quality.

Support

This work was supported by the Korea Research Foundation Grant funded by the Korea Govement (MOE‐HRD)” (KRF‐2006‐331‐E00235).