bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Flap Tension in Midface Surgery
Authors: Herrero Nicholas A , Ethan Sherman , Adelson Robert T.
Published: August 2008

Problem

Despite a large volume of literature addressing rejuvenation of the midface, there is an absence of objective data addressing the various approaches to midface lifting. This study compares flap tensions generated by midface lifts though transblepharoplasty (TB) and transtemporal (TT) surgical approaches, with and without the additional intervention of a transoral periosteal release.

Methods

12 fresh cadaver heads underwent standard TB and TT approaches for midface lifting with Endotine (Coapt Systems, Inc, Palo Alto, CA) fixation devices. A machined apparatus, designed for this experiment, pulled the midface flap one centimeter along a posterior‐superior vector while a precision digital load cell recorded the generated tension (grams). A transoral periosteal release was performed and flap tensions were measured again after this intervention. Green‐house‐Geisser test allowed analysis of the three‐way interaction between surgical approach, periosteal release, and the number of times the test was repeated.

Results

Flap tension decreased significantly during testing with each sequential repeated pull on the midface (p<0.0001). Average flap tension for TB approach did not differ significantly from that of TT approach, whether or not a periosteal release was performed. However, while no particular approach was found to have a significant reduction in flap tension, a transoral periosteal release produced a significant reduction in flap tension (12%) when compared to trials without periosteal release (p=0.0191).

Conclusion

A significant reduction in flap tension as a result of transoral periosteal release is demonstrated in both transtemporal and transblepharoplasty approaches for midface lifting. This objective reduction in flap tension may allow the surgeon greater mobility of the midface tissues and perhaps less tension opposing flap fixation.

Significance

The objective reduction in flap tension by employing a transoral subperiosteal release may allow the surgeon greater mobility of the midface tissues and perhaps less tension opposing flap fixation.

Support

Coapt Systems, Inc, (Palo Alto, CA) provided the endotine devices utilized in the experiment.
Electrical Pacing of Paralyzed Orbicularis Oculi Muscle
Authors: Sanderson Jeremy Drew , Otto Randal A
Published: August 2008

Problem

In this study, the authors investigated the feasibility of restoring function by direct electrical stimulation of the paralyzed orbicularis oculi muscle in the dog model.

Methods

Seven mongrel dogs underwent bilateral surgical resection of a 1cm portion of their proximal facial nerves followed by bilateral placement of an electrical pacing device with electrodes placed directly into the orbicularis oculi muscles. One eye was stimulated and the other used as a control. The eye was stimulated for 24 hours daily for 90 days. The function of the electrical stimulation in reproducing a normal protective blink was then graded on a scale of 0 to 3. The worst grade of 0 was assigned if no movement or only the slightest twitch could be visualized in the eyelid, a grade of 1 with slight closure, 2 for a protective blink and 3 for overclosure.

Results

A total of 22 eyelids were stimulated and graded in the 7 dogs. The overall average blink grade in the stimulated eyes was 1.7 and the range of the averages was from 0.06 to 2.8. An average blink grade in all the unstimulated eyes was 0. Thirty‐two percent of the stimulated eyelids obtained a blink considered to be protective or obtained over closure corresponding to a blink grade of 2–3.

Conclusion

Although current spread leading to a poor functional result was a common problem, the results when this complication could be overcome were very encouraging. To our knowledge this represents the first report of functional electrical stimulation of the paralyzed orbicularis oculi muscle on a continuous basis for 90 consecutive days.

Significance

The findings of this study support the feasibility of employing direct electrical stimulation to restore the function of the orbicularis oculi muscle.

Support

This study was funded through a grant from Medtronics, Inc (Minneapolis, MN)
Expression of VEGF and VEGF Receptors in Port‐Wine Stains
Authors: Ramakrishnan Jeevan B , Fan Chun‐Yang , Cetin Neslihan , Buckmiller Lisa M , Vural Emre A.
Published: August 2008

Problem

Port‐wine stains (PWS) are vascular anomalies that may cause significant functional and psycho‐social ramifications worsening by age. Traditional management of port‐wine stains involves pulsed dye laser treatments, however only 10% of these lesions show complete response and 35% of these recur even after complete response. Therefore, there is still a need for an effective treatment for PWS. Vascular endothelial growth factor (VEGF) ‐a potent angiogenic molecule‐has been shown to be associated with various vascular anomalies such as hemangiomas and arterio‐venous malformations. However, the possible pathophysiological relationship of VEGF and PWS has not been investigated previously. The purpose of this study is to investigate if VEGF and VEGF‐receptors are overexpressed in PWS as a key step in developing a potential targeted treatment approach for this disfiguring disease.

Methods

12 surgically resected PWS specimens and 12 normal skin specimens were immunohistochemically stained by using anti‐VEGF and anti‐VEGF receptor 2 antibodies.

Results

PWS specimens revealed significantly higher levels of expression of both VEGF molecule and VEGF‐receptor 2 than the control specimens.

Conclusion

VEGF and VEGF‐receptor 2 may play a role in pathophysiology of PWS. Overexpression of these molecules may suggest that PWS could have proliferating potential in addition to their hypertrophic growth pattern. Therefore, targeted approaches against VEGF and its receptors may have a potential role in the treatment of these disfiguring lesions.

Significance

1. Expression of VEGF and VEGF receptors in PWS has not been investigated previously. 2. The results of this study may suggest that PWS could have proliferating potential, in contrast to the common belief that PWS progress by hypertrophy rather than hyperplasia. 3. Targeted approaches against VEGF and its receptors may have a potential role in the treatment of these disfiguring lesions.

Support

This study was supported by Medical Research Endowment Award at university of Arkansas for Medical Sciences.
Stem Cells from New Sources for Cartilage Tissue Engineering
Authors: Goessler Ulrich R. , Stern‐Straeter Jens , Bran Gregor , Sadick Haneen , Hoermann Karl , Riedel Frank
Published: August 2008

Problem

The use of adult mesenchymal stem cells (MSC) – especially from new sources including adipose tissue ‐ offers new perspectives in the generation of transplants for reconstructive surgery. The extracellular matrix (ECM) plays a key role in modulating function and phenotype of the embedded cells and contains the integrins as adhesion receptors mediating cell‐cell‐ and cell‐matrix‐interactions. In our study, characteristic changes in integrin expression during the course of chondrogenic differentiation of MSC from bone marrow and adipose tissue were compared.

Methods

MSC were isolated from bone marrow biopsies and adipose tissue. During cell culture, chondrogenic differentiation was performed. The expression of Integrins and their signaling components were analyzed with microarray and immunohistochemistry in freshly isolated MSC and after chondrogenic differentiation.

Results

The Fibronectin‐Receptor (Integrin a5b1) was expressed by undifferentiated MSC, expression rose during chondrogenic differentiation in both types of MSC. The components of the Vitronectin/Osteopontin‐Receptors (avb5) were not expressed by freshly isolated MSC. Expression rose with ongoing differentiation. Receptors for Collagens (a1b1, a2b1, a3b1) were weakly expressed by undifferentiated MSC and were activated during differentiation. As intracellular signaling components integrin linked kinase (ILK) and CD47 showed increasing expression with ongoing differentiation. For all integrins, no significant differences could be found in the 2 types of MSC.

Conclusion

Integrin‐mediated signaling seems to play an important role in the generation and maintenance of the chondrocytic phenotype during chondrogenic differentiation. The receptors for Fibronectin, Vitronectin, Osteopontin and Collagens in particular might be involved in the generation of the ECM. Intracellularly, their signals might be transduced by ILK and CD47.

Significance

To fully harness the potential of these cells, future studies should be directed to ascertain their cellular and molecular characteristics for optimal identification, isolation, and expansion.
TGF‐beta1 Targeting on MMP—Expression in Keloid Fibroblasts
Authors: Sadick Haneen , Goessler Ulrich R. , Stern‐Straeter Jens , Hoermann Karl , Riedel Frank
Published: August 2008

Problem

Transforming growth factor‐beta1 (TGF‐beta1) has been identified as an important regulator of wound healing. Recent developments in molecular therapy offer exciting prospects for the modulation of wound healing, specifically those targeting TGF‐beta1. The purpose of this study was to analyze the effect of TGF‐beta1 targeting on the expression of matrix metallo‐proteinases (MMP‐2 and MMP‐9) in fibroblasts cultured from earlobe keloids.

Methods

The expression of MMP‐2 and MMP‐9 in tissue samples from keloids was investigated by immunohistochemistry. The effect of TGF‐beta1 targeting using antisense oligonucleotides on the expression of MMPs in keloid‐derived fibroblasts was analyzed by ELISA and multiplex RT‐PCR.

Results

Immunohistochemical studies demonstrated an increased expression of MMP‐2 and MMP‐9 proteins in tissue samples from keloids compared to normal human skin. The treatment of human keloid‐derived fibroblasts with TGF‐beta1 antisense oligonucleotides (ONDs) in vitro efficiently down‐regulated MMP‐9 expression but not the MMP‐2 expression. In the supernatans of keloid‐derived fibroblasts, the TGF‐beta1 antisense therapy showed a decreased secretion level of both MMP‐9 and MMP‐2. So other than in MMP‐9, the synthesis of MMP‐2 does not seem to be influenced by TGF‐beta1 anti‐sense ONDs whereas the secretion of MMP‐2, similar to that of MMP‐9, is significantly decreased in the supernatans of keloid‐derived fibroblasts.

Conclusion

TGF‐beta1 antisense OND technology may be a potential therapeutic option for the inhibition of proteolytic tissue destruction in keloids. The results should be regarded as a further implication for the possible treatment of keloids.

Significance

The results should be regarded as a further implication for the possible treatment of keloids.
A Dynamic 3D Model of Hyoid Muscle Groups
Authors: Zu Yihe , Perlman Adrienne L , Sutton Bradley P , Sinn‐Hanlon Janet Marie , Yang Zhenyu
Published: August 2008

Problem

A 3D dynamic model of supra and infrahyoid muscle groups was developed to provide information relative to the extent of muscle contraction. This model can provide valuable information relative to the physiology and pathophysiology of speech production, swallowing, and effects of surgical reconstruction. The rotation of the 3D model permits the viewer to appreciate the internal anatomy from several vantage points. Transparency can be used to study underlying anatomy without change the surroundings. In this study, a 3D dynamic model of suprahyoid and infrahyoid muscle groups during swallowing was created based on still 3D and dynamic MRI. This model will be further developed to examine the effects of muscle forces on functional changes after head and neck surgery.

Methods

3D MRI of head and neck area were used to reconstruct 3D polygonal meshes of hyoid muscle groups, hyoid bone, mandible, and larynx. Measurement was performed using Amira 4.1. The polygonal meshes were imported in to Alias Maya 8.5. By mapping the dynamic MRI of the swallow, the muscle meshes were reshaped to generate a 3D movie of swallowing.

Results

Using MR images, an accurate and realistic computer reconstruction of supra and infra hyoid muscles, in situ, was constructed. A 3D movie was created to demonstrate the anatomy, larynx, and hyoid displacement in all directions.

Conclusion

MRI provides high quality soft tissue information. 3D modeling using MRI provide muscle information in situ. The dynamic movie created the opportunity for researchers and physicians to observe anatomical changes in all coordinate planes. These offer a promising future toward increased understanding of head and neck physiology and pathophysiology.

Significance

This is the first 3D model to provide dynamic information on the hyoid muscle groups. It will eventually include forces that will animate functional change after surgical reconstruction.

Support

NIH‐NIDCD, Award No. PHS 1 R01 Dc005603‐01A2.
A Multiparametric Gene‐Based Diagnosis Model for HNSCC
Authors: Worsham Maria J , Lu Mei , Sethi Seema , Kapke Alissa , Chen Kang‐Mei , Havard Shaleta E , Benninger Michael S
Published: August 2008

Problem

A current shortcoming in the diagnosis, prognosis and treatment of HNSCC is a lack of methods that adequately addresses the complexity and diversity of the disease. Diagnostic and prognostic marker systems based on single parameters have generally proven inadequate. Multiparametric methods, which rely on many pieces of information, are ideally suited to the grouping of tumor subtypes, identification of specific patterns of disease progression, and in predicting clinical outcomes.

Methods

In a retrospective multi‐ethnic primary HNSCC cohort drawn from a primary healthcare setting, and constructed through re‐review of the primary biopsy, gene alterations (104 genes) and clinical variables (9 histopathology and 3 demographic variables) were evaluated as predictors of stage (TNM, early versus late). Statistical analysis compared logistic regression and Classification and Regression Tree (CART®) analyses to derive the most predictive model, assessed using receiver operating characteristic (ROC) curve analysis.

Results

Considering all clinical and gene variables for the 360 primary HNSCC study cohort, the multivariate logistic regression model retained only tumor grade, sample type (radical dissection) and their interaction with ROC as 64%. CART® generated a multivariable model with 12 variables: clinical variables of age, pattern of invasion, and tumor grade and gene variables of TP53, F3, TFF1, CDKN2A, KIAA0170, HS222808, TANK, MYC, and UTY1, with an ROC of 0.82%.

Conclusion

A group of clinical variables in multiparametric combinations with molecular alterations discriminated early and late stage HNSCC. CART® improved the model's performance.

Significance

Validation of this initial multiparametric strategy for predicting late stage HNSCC comprising several genes and clinical factors, currently underway, should yield a multiparametric, comprehensive genome‐wide molecular algorithm integrated with clinical risk factors in order to refine HNSCC diagnosis and prognosis associated with clinical and pathological staging to aid in the clinical management of patients at the earliest stages.

Support

NIH R01 DE15990.
Abrogation of PDGFR by STI571: Effect on PDGF/VEGF in HNSCC
Authors: Bran Gregor , Stern‐Straeter Jens , Goessler Ulrich R. , Bran Birgit , Hoermann Karl , Riedel Frank
Published: August 2008

Problem

Angiogenesis is increased in head and neck squamous cell carcinoma (HNSCC), and correlates with tumor progression and metastasis. Vascular endothelial growth factor (VEGF) has been shown to be a key regulator of angiogenesis. Microvascular density is known to correlate with metastasis and aggressiveness. PDGF and PDGFR are expressed by many tumor types, including HNSCC. The PDGF/PDGFR‐System is believed to play a pivotal role in tumorigenesis. The compound STI571 (Imatinib mesylate, Gleevec) inhibits PDGFR tyrosine kinases. In this study, we seek to determine whether targeting PDGFR signalling in HNSCC cell lines may affect expression and secretion of PDGF as well as VEGF in vitro.

Methods

Established human HNSCC cell lines were incubated with STI571 of different concentrations for 24h and 72h. PDGF and VEGF concentrations were determined by an ELISA technique, and gene expression was analyzed by RTPCR.

Results

In vitro treatment of HNSCC cell lines with STI571 resulted in a significant reduction of expression and secretion of PDGF and VEGF. We also observed a dose related effect for VEGF synthesis.

Conclusion

The results in this study suggest, that inhibition of PDGFR using STI571 might have an antitumor effect mediated in part by inhibition of VEGF‐induced angiogenesis in HNSCC. Further studies need to be conducted to improve understanding of the molecular mechanisms.

Significance

Our study indicates that STI571 might play a role in establishing new therapeutic strategies against HNSCC.
Complementary Test of Antibodies to EBNA‐1 and EA for NPC
Authors: Chang Kai‐Ping , Hao Sheng‐Po
Published: August 2008

Problem

This hospital‐based cohort study evaluated the efficacy of three Epstein‐Barr virus (EBV) ‐ associated assays for nasopharyngeal carcinoma (NPC) primary screening and monitoring treatment outcome.

Methods

Five hundred seventeen consecutive subjects, including 156 NPC patients, 264 healthy volunteers and ninety‐seven patients with head‐and‐neck squamous cell carcinoma (HNSCC) were enrolled. The sensitivity and specificity of EBV IgAs to viral capsid antigen (VCA), complementary EBV IgAs to early antigen and nuclear antigen‐1 (EA+EBNA‐1), and EBV DNA load were examined by immunofluorecent assays, enzyme‐linked immunosorbent assays, and quantitative real‐time PCR, respectively.

Results

After constructing the receiver operating characteristics to demonstrate screening efficacy, EBV EA+EBNA‐1 IgA (AUC: 0.952; 95% CI, 0.930–0.974) was proved superior to EBV VCA IgA (AUC: 0.888; 95% CI, 0.854–0.922) or EBV DNA load (AUC: 0.893; 95% CI, 0.854–0.932) in differentiating NPC patients from controls. Comparison of screening efficacy between NPC patients and HNSCC patients revealed EBV EA+EBNA‐1 IgA (AUC: 0.964; 95% CI, 0.943–0.985) still outperformed EBV VCA IgA (AUC: 0.884; 95% CI, 0.845–0.923). In subjects with higher serum titer or level equal to or above 1:80 and 6 EU/ml for EBV VCA IgA and EA+EBNA‐1 IgA, the specificity reached as high as 99.2% and 95.1%, respectively, in the control groups. However, correlation of these three assays with clinicopathological manifestations of NPC, revealed only EBV DNA load significantly associated with N stage and overall stage in NPC patients. Additionally, EBV DNA load could be used to further raise the specificity of EBV EA+EBNA‐1 IgA assays and was also the only assay to be consistently predictive of tumor relapse in post‐treatment patients according to serial test results by time frame.

Conclusion

The EBV EA+EBNA‐1 IgA‐based protocol is recommended for mass screening, but EBV DNA load should be used solely for post‐treatment monitoring for NPC in endemic areas.

Significance

A possible alternative for primary screening of NPC.

Support

NSC of Taiwan, ROC. (NSC92‐2314‐B‐182A‐207) and CGU (CMRPG32090 and CMRPG360211).
IL10, An Independent Predictor of Outcome in HNSCC Patients
Authors: Alhamarneh Osama , Stafford Nicholas D. , Greenman John
Published: August 2008

Problem

To determine the correlation between IL10, a Th2‐type inhibitory cytokine, clinical outcome and survival in HNSCC patients.

Methods

IL10 levels in the serum of newly‐presenting, untreated, patients with HNSCC were measured pre‐treatment (n=107) and 4–6 weeks after treatment (n=43), and were compared with a cohort of healthy controls (n=40) of similar age and sex. A commercial IL10 ELISA (Biosource) with a minimum detectable level of 0.2 pg/ml was used. Statistical analysis of associations between the levels and detectability of IL10 and clinical outcome and survival were done.

Results

Both IL10 detectability and levels were significantly higher in patients than in controls (p=0.001). Post treatment, IL10 levels dropped significantly (p=0.02). Pretreatment, IL10 levels were significantly elevated in the advanced stage of the disease (III/IV vs. I/II), in node positive patients and in patients with bulkier tumor load (T3/T4 vs. T1/T2); p=0.005, 0.037 and 0.001 respectively. The larynx (n=36), oropharynx (n=25) and pharynx (n=16) showed significantly higher levels and increased detectability of IL10 in the pre‐treatment group when compared to the post treatment group, however, oral cavity tumors (n=21) showed the opposite. Finally, the detectability of IL10 significantly correlated with poorer survival (Kaplan‐Meier, p=0.026) after a mean follow up of 15 (range 1–36) months.

Conclusion

IL10 levels drop significantly once the tumor mass is removed suggesting that this is the most important source of the circulating cytokine. IL10, as well as the tumor bulk, the nodal status and the overall tumor stage, were shown to be independent factors in predicting a poorer clinical outcome and worse survival in tumors originating in the larynx, pharynx and oropharynx, but not oral cavity, suggesting distinct inter‐tumour differences.

Significance

IL10 could play a potential role as a prognostic marker in HNSCC, in addition to the possible manipulation of IL10 in future immunotherapeutic agents.
Immunomodulation of Vitamin D in Head and Neck Cancer
Authors: Kulbersh Jonathan S , Day Terry A , Gillespie Marion B. , Young M Rita
Published: August 2008

Problem

Quantitate the intratumoral differentiation of HNSCC induced immunoinhibitory CD34+ in response to enteral treatment of 1,25 dihydroxyvitamin D3 (1,25D3).

Methods

8 patients with HNSCC were treated orally with 3 weeks of 1,25D3 before surgical excision. Tumor tissue was collected and analyzed using immunohistochemistry to test the hypothesis that treatment would drive differentiation of immune inhibitory CD34+ cells into immune stimulatory dendritic cells. Patients’ clinical data for high risk pathological features were also analyzed.

Results

After 1,25D3 treatment, intratumoral levels of CD34+ cells significantly dropped. Levels of DC‐lamp, a cellular marker for mature dendritic cells, significantly increased while there was not a change in intratumoral levels immature dendritic cell marker DC‐sign. Levels of CD4+ cells were unchanged while levels of CD8+ cells were significantly decreased. There were no associated statistically significant correlations between CD34+ levels and histological features, although there was a trend of increasing cervical metastasis in patients with increased intratumoral CD34+ cells.

Conclusion

CD34+ cells are hematopoietically immature cells that have been shown to differentiate to dendritic cells when grown in culture. This is the first study to analyze the effects of enteral 1,25D3 on intratumoral immune infiltrate, specifically CD34+ cells. 1,25D3 stimulated differentiation of CD34+ immunosuppressive cells to differentiate into immunostimulatory dendritic cells.

Significance

Despite recent advances in treatment, patients with HNSCC have a five year mortality rate of less than 50%. One aspect contributing to the poor prognosis is the profound immunoinhibitory effects induced by HNSCC. It has been shown that CD34+ suppressor cells are a component of this immunosuppresion induced by the carcinoma. Adding 1,25D3 in the treatment of HNSCC decreased levels of intratumoral CD34+ cell population and diminish the profound immunoinhibitory effects of the cancer. These studies support the use of 1,25D3 as a neo‐adjunctive therapy to overcome HNSCC profound immunosuppression thereby potentiating other promising immunotherapies.

Support

Clinical Sciences Research Service, Department of Veterans Affairs, Washington, D.C. Vitamin D therapy to enhance intratumoral anti‐cancer immune reactivity October 2004 ‐ September 2009 $747,000; University Research Committee Resident Research Grant, Medical University of South Carolina January 2008 $2,000.
Molecular Classification of HPV‐Associated Head Neck Cancer
Authors: Weinberger Paul M. , Yu Ziwei , Kountourakis Panteleimon , Sasaki Clarence T , Kowalski Diane , Rimm David , Camp Robert L , Amanda Psyrri
Published: August 2008

Problem

We previously proposed that head and neck squamous cell carcinoma (HNSCC) presents in three histopathologically similar but molecularly distinct types where p16 expression combined with high‐risk Human Papillomavirus (HPV) presence defines three distinct subgroups: Class I (HPV‐), Class II (HPV16+/p16‐), Class III (HPV16+/p16+). In this study we sought to further explore and validate this classification schema.

Methods

Paraffin‐embedded specimens from 77 patients with OSCC classified into the 3‐class model based on p16 expression and HPV DNA presence were previously queried for expression of 14 target proteins with known involvement in tumor progression using AQUA(tm). We calculated pairwise associations by Spearman correlation and performed hierarchical clustering analysis to assess global expression patterns of class I, II and III tumors.

Results

Hierarchical clustering analysis demonstrated class I and class II tumors displayed similar clustering patterns. Class III tumors (HPV+/p16+) displayed unique clustering patterns distinct from both class I and class II tumors. In class I/II tumors ERK2, p27, p14, ki67 and PCNA were tightly associated. This contrasted with class III tumors which displayed two separate clusters (EGFR, RB, pAKT and CND1), and (ERK2, p16 and p14).

Conclusion

We demonstrated that HNSCC classified by a combination of HPV DNA presence and p16 expression status have distinct molecular phenotypes. Class III tumors, distinct from HPV+ but p16‐negative class II tumors, have a unique molecular clustering profile, lending support to the proposed 3‐class model.

Significance

These findings may have significant implications in patient selection for HPV‐specific therapies.

Support

AAO/HNSF Resident Research Grant 26357 (PMW).
Molecular Markers of Micrometastasis in OCSCCA
Authors: Germani Ross M , Civantos Francisco J , Franzmann Elizabeth J
Published: August 2008

Problem

To report our experience in analysis of immunohistochemical markers for detection of micrometastasis in patients with oral cavity squamous cell carcinoma (OCSCCA)who have undergone lymphoscintigraphy and sentinel lymph node biopsy(SLNB). Staining results and analysis for CD‐44, MT1‐MMP, FAK and E‐cadherin will be presented.

Methods

Paraffin embedded specimens (N=30) were retrospectively studied by immunohistochemical analysis. Applied monoclonal antibodies were anti‐CD‐44(Bender Med‐Systems Inc., Burlingame, CA), anti‐MT1‐MMP(Calbiochem, San Diego, CA), anti‐FAK (Upstate Biotechnology Inc, Lake Placid, NY) and anti‐E‐cadherin (Transduction Laboratories, Lexington, Ky). Staining patterns were analyzed by 2 independent pathologists. Differences between groups for each staining pattern were analyzed by the Fisher exact method.

Results

2 groups were identified based on results of SLNB: patients with negative sentinel lymph node biopsies who represented true N0 necks (N = 19; group I) and those with evidence of occult lymph node metastasis (N =11; group II). There was no significant difference in staining patterns between groups for CD‐44, although three distinct staining patterns were identified. For MT1‐MMP there was a striking difference in staining between groups I (20%) and II (67%) that did not reach statistical significance (p=0.13). Specimens are currently undergoing staining for FAK and E‐cadherin as described. Staining results for these markers will be analyzed and compared to those for CD‐44 and MT1‐MMP.

Conclusion

Lymphoscintigraphy followed by SLNB has been established as a sensitive tool to detect early tumor metastasis to proximal nodal basins. By comparing sentinel node status with molecular events at the primary tumor site we hope to identify a molecular marker for metastasis. We will report results of immunohistochemical analysis for CD‐44, MT1‐MMP, FAK and E‐cadherin in this manuscript.

Significance

By studying this unique subset of patients we hope to identify a reliable molecular marker for metastasis, which may ultimately change the way we manage the neck for early oral cavity cancer.

Support

Resident research stipend.
Molecular Similarity of HNSCC from Different Subsites
Authors: Weinberger Paul M , Merkley Mark A , Jeffrey Lee , Gourin Christine G , Podolsky Robert , Sasaki Clarence T , Amanda Psyrri , Dynan William
Published: August 2008

Problem

Squamous cell carcinomas arising from various subsites within the head and neck (HNSCC), while histologically identical, have substantial differences in survival and recurrence rates. Controversy exists as to whether this reflects physical differences or fundamental molecular heterogeneity between subsites. In this study, we used two proteomics approaches to evaluate HNSCCs for differences in protein expression between subsites including oral cavity, oropharynx, larynx and hypopharynx.

Methods

Tumor tissue from a total of 85 patients with HNSCC was queried by two complementary proteomics techniques. Frozen tissue from 14 patients with histologically confirmed HNSCC was enriched for tumor and normal tissue by laser capture microdissection. Total protein was extracted, separated by 2D‐difference gel electrophoresis (2D‐DIGE), and evaluated for differential protein expression between subsites. A tissue microarray (TMA) was constructed consisting of 71 patients with HNSCC. This TMA was queried for expression of 4 cell‐cycle and regulatory proteins chosen a priori for their known roles in cancer, using Automated Quantitative Analysis of protein expression (AQUA).

Results

From the first cohort, proteins from tumor and matched normal tissue were successfully isolated and 732 spots identified as matching across >90% of gels. After supervised statistical analysis, we identified no significant differences in protein expression between subsites. AQUA analysis of the second, larger cohort likewise revealed no difference between subsite for cyclin D1, p53, Rb, or p14 expression.

Conclusion

Using two validated, complementary proteomics methods we identified no significant differences between HNSCCs from different head and neck subsites.

Significance

It is possible that heterogeneity among HNSCCs may be based not on anatomic subsite, but rather on fundamentally different carcinogenic pathways – i.e. virally induced versus carcinogenic insult (tobacco, alcohol).

Support

NIHR33‐CA‐5941(WSD); AAO/HNSF Resident Research Grant 26357 (PMW).
Reirradiation After Surgery and Microvascular Reconstruction
Authors: Suh Jeffrey D. , Kim Brian Paul , Abemayor Elliot , Sercarz Joel A , Nabili Vishad , Liu Jerome H , Juillard Guy Jf , Blackwell Keith E
Published: August 2008

Problem

To evaluate the outcome and complications of reirradiation of recurrent head and neck cancer after salvage surgery and microvascular reconstruction.

Methods

Retrospective Study. Twelve patients underwent salvage surgery with microvascular reconstruction for recurrent or new primary head and neck cancer in a previously irradiated field. Median prior RT dose was 63.0 Gy (range 30.0–72.8). Patients then underwent postoperative reirradiation, receiving a median total cumulative radiation dose of 115.0 Gy.

Results

Three patients (25%) experienced acute complications (<3 months) during reirradiation that resolved with conservative care. Four patients (33%) developed grade 3 or 4 late reirradiation complications (>3 months). There were no incidences of free flap failure. No patients suffered brain necrosis, spinal cord injury, or carotid rupture. The incidence of soft tissue necrosis and osteoradionecrosis was 8%. There were no treatment‐related mortalities. Six patients (50%) are alive without evidence of recurrent disease a median of 40 months after reirradiation (range 4–64 months).

Conclusion

Free flap reconstruction followed by reirradiation is not associated with an increased risk of perioperative, acute, or late complications. Microvascular free flaps allow for maximal resection and reliable reconstruction of previously irradiated cancers before high dose reirradiation, and may reduce the incidence of severe late complications and treatment related mortality.

Significance

Reirradiation for recurrent head and neck squamous cell carcinoma remains controversial. However, increasing evidence has demonstrated improved survival and locoregional control with reirradiation at the cost of potentially severe or sometimes fatal radiation toxicity. We hypothesize that using well‐vascularized tissue and bone at the time of salvage surgery can reduce the incidence of reirradiation complications. This would allow patients at high risk for recurrence to more safely receive a second course of radiation therapy. To our knowledge this is the first report of the effects of microvascular reconstruction on complications and outcomes of patients undergoing salvage surgery and external beam reirradiation.
Targeted Liposomal Paclitaxel Raises Tumor Vessel Leakiness
Authors: Strieth Sebastian , Eichhorn Martin E , Alexander Werner , Sauer Birgitta , Teifel Michael , Michaelis Uwe , Berghaus Alexander , Dellian Marc
Published: August 2008

Problem

Paclitaxel encapsulated in cationic liposomes (EndoTAG‐1) is a vascular targeting formulation for the treatment of solid tumors. It triggers intratumoral microthrombosis causing significant inhibition of tumor perfusion and tumor growth associated with endothelial cell apoptosis. Here, we quantified the effects of repeated EndoTAG‐1 therapy on tumor microvascular leakiness with respect to leukocyte‐endothelial cell interactions, the targeting property of cationic liposomes and the therapeutic combination with conventional cisplatin chemotherapy.

Methods

Using dorsal skinfold chamber preparations in Syrian Golden hamsters in vivo fluorescence microscopy experiments were performed after repeated EndoTAG‐1 treatment of A‐Mel‐3 tumors. Controls received glucose, paclitaxel alone or cationic liposomes devoid of paclitaxel. Extravasation of rhodamine‐labelled albumin was measured to calculate microvessel permeability and intratumoral leukocyte‐endothelial cell interactions were quantified. Subcutaneous tumor growth was evaluated after combination therapy followed by histological analysis.

Results

Microvascular permeability was significantly increased only after treatment with EndoTAG‐1, while intratumoral leukocyte‐endothelial cell interactions were not affected by any treatment. In separate skinfold chamber experiments fluorescently‐labeled cationic liposomes kept their targeting property for tumor endothelial cells after repeated EndoTAG‐1 treatment and no signs of extravasation were observed. Subcutaneous A‐Mel‐3 tumor growth was significantly inhibited by the combination of cisplatin and EndoTAG‐1.

Conclusion

These data show that vascular targeting with EndoTAG‐1 increases tumor microvessel leakiness probably due to vascular damage. This mechanism is not mediated by inflammatory leukocyte‐endothelial cell interactions. Manipulating the blood‐tumor barrier by repeated tumor microvessel targeting using EndoTAG‐1 can effectively be combined with tumor cell‐directed conventional cisplatin chemotherapy.

Significance

Antivascular therapy using liposomal paclitaxel (EndoTAG‐1) is currently undergoing Phase II clinical studies in human cancer patients. Referring to the present experimental data showing increased permeability of intratumoral microvessels combination therapy with conventional therapy appears to be a very promising strategy.

Support

This study was supported by grants of Munich Biotech AG, Neuried, Germany and by grants of the Novartis Foundation for Therapeutic Research, Nürnberg, Germany.
Understanding EMT in HNSCC
Authors: John Maie St
Published: August 2008

Problem

Down regulation of E‐cadherin is the hallmark of epithelial mesenchymal transition (EMT). The loss of E‐cadherin confers a more invasive phenotype and is one of the steps leading to metastasis. Our recent investigations indicate that inflammatory mediators are potent regulators of EMT in HNSCC.

Methods

Immunohistochemical analysis of HNSCC tumor tissue specimens, in vitro analysis of E‐cadherin regulation in HNSCC cells, and in vivo analysis of tumor growth and metastasis ina murine model.

Results

An inverse relationship between COX‐2 and E‐cadherin was demonstrated in situ by double immunohistochemical staining of human HNSCC tissue sections. Treatment of HNSCC cells with exogenous IL‐1B, also caused the down regulation of E‐cadherin expression and up regulation of Cox‐2 expression in HNSCC cell lines in a concentration‐ and time‐dependent manner. IL‐1B ‐treated HNSCC cell lines demonstrated a significant decrease in E‐cadherin mRNA and an increase in the mRNA of the transcriptional repressors SNAIL and ZEB1. IL‐1B exposure led to enhanced ZEB1 and Snail binding at the chromatin level as determined by chromatin immunoprecipitation assays (ChIP). An inverse relationship between E‐cadherin and ZEB1 and a direct relationship between COX‐2 and ZEB1 were demonstrated by immunohistochemical staining of human HNSCC tissue sections. When HNSCC cell lines that over express Snail were injected into SCID mice, the primary tumor and metastatic burdens were significantly greater than HNSCC vector controls.

Conclusion

These findings indicate that 1L‐1B modulates transcriptional repressors of E‐cadherin and thereby regulates COX‐2‐dependent E‐cadherin expression in HNSCC. This is the first report indicating the role of E‐cadherin transcriptional repressors in the inflammation‐induced promotion of EMT in HNSCC.

Significance

This newly defined pathway for transcriptional regulation of E‐cadherin in HNSCC has important implications for targeted chemoprevention and therapy.

Support

AA0–HNS Surgeon Scientist CDA.
Effect of Allergens and Tobacco Smoke on Laryngeal Mucosa
Authors: Belafsky Peter C , Mouadeb Debbie A. , Pinkerton Kent , Birchall Martin A , Konia Thomas
Published: August 2008

Problem

Environmental factors such as second hand smoke and allergens are known to play an important part in the development of lower airway disease, specifically asthma. The effect of these common exposures on the larynx has been neglected. The purpose of this investigation was to investigate the effects of environmental tobacco smoke (ETS) and allergens on laryngeal mucosa.

Methods

Seventeen guinea pigs were exposed by inhalation to: 1) air and saline, 2) air and house dust mite allergen (HDMA), 3) ETS and saline 4) HDMA and ETS. Exposures began at 3 weeks of age and lasted 10 weeks. Histology was evaluated in different subsets of the larynx for eosinophils and mucin, markers of inflammation.

Results

Eosinophils in the supraglottis were elevated in the air/HDMA and ETS/HDMA groups (p<0.05). Animals exposed to HDMA were 4 times more likely to have > 10 eosinophils per HPF in the supraglottis than control animals (95% CI = 0.74, 20). Trends towards mucin hypersecretion in the subglottis of the ETS/air and ETS/HDMA groups were noted. Animals exposed to ETS and HDMA were 3.8 times more likely to have 25% of the subglottic epithelium stain PAS positive for mucin (95% CI =. 59, 23.66).

Conclusion

The data suggest that environmental tobacco smoke and inhaled allergens contribute to the pathogenesis of laryngeal inflammation.

Significance

Environmental tobacco smoke and allergen exposure may play a role in the development of chronic laryngitis.
Localization of Phonation Integration Center in Rabbits
Authors: French Lesley C. , Ohno Tsunehisa , Omid Majdani , Zealear David L , Labadie Robert F , Rousseau Bernard
Published: August 2008

Problem

The midbrain periaqueductal gray (PAG) serves as a phonation integration center, coordinating respiratory, laryngeal, and orofacial activity during vocalization. The objective of this pilot study was to explore electrical excitation of the midbrain PAG for eliciting vocalization using an in‐vivo rabbit preparation and determine the feasibility of using image guidance for improved localization of target midbrain structures.

Methods

Pilot experiments were conducted using four New Zealand White (NZW) breeder rabbits. In the first experiment, a stereotaxic frame was used to guide stimulation of the midbrain PAG without the use of image guidance. Decisions regarding track placements were directed by a stereotaxic atlas of the NZW rabbit brain. Histologic stains of axial brain sections were then obtained to determine electrode track positioning. In a subsequent experiment, magnetic resonance imaging (MRI) and computed tomography (CT) scans were obtained and used to localize midbrain structures for advanced planning capabilities.

Results

Without the use of image guidance, phonation was feasible, measuring 80–100 Hz using electrical excitation of the midbrain PAG (0.5 to 1.5 mA, 200 Hz, 2 second train duration). However, precise localization of the midbrain PAG proved technically challenging. Using CT and MRI data, scans could be loaded onto an image guided surgical system and combined using an image fusion tool for advanced planning.

Conclusion

The authors demonstrate the feasibility of electrical excitation of the midbrain PAG to produce vocalization in rabbits using a stereotaxic brain atlas and offer an alternative approach using image guidance for advanced planning and more precise localization of target midbrain structures.

Significance

The biomechanical forces of phonation and dynamic nature of vocal fold remodeling have created a need for chronic animal preparations to investigate these reparative processes in‐vivo. Image guided techniques may improve the feasibility of brain stimulation techniques for eliciting and studying time dependent changes in vocal fold remodeling.

Support

NIH‐NIDCD R03 DC008400.
Nerve‐Induced Rabbit Phonation: Temporal Phonation Stability
Authors: Rousseau Bernard , Ge Pingjiang , French Lesley C , Ohno Tsunehisa , Zealear David L , Ossoff Robert H
Published: August 2008

Problem

In‐vivo phonation using animals with layered structure similarities and phonation characteristics akin to humans are needed for the study of vocal fold reparative processes. The authors’ previous studies have demonstrated the feasibility of eliciting vocalization in a flow‐controlled in‐vivo rabbit preparation. In the current study, the authors provide information relative to temporal stability of nerve elicited rabbit phonation.

Methods

Eight New Zealand White breeder rabbits received pulsatile current to the laryngeal nerves with airflow delivered at a constant rate. Measurements of vocal intensity and fundamental frequency were recorded every 5, 30, and 90 minutes after the onset of phonation. Pearson correlations were used to investigate relationships among vocal intensity and fundamental frequency.

Results

Phonation was elicited with fundamental frequency and intensity ranging from 300 to 1000 Hz and 68 to 80 dB SPL, respectively at 5 minutes, 55 to 840 Hz and 70 to 79 dB SPL at 30 minutes, and 51 to 438 Hz and 68 to 76 dB SPL measured at 90 minutes. Correlation coefficients revealed predictable relationships among the dependent variables over time.

Conclusion

Continuous, audible phonation within the range of human phonation was produced over a 90 minute duration using an in‐vivo rabbit preparation. Expected relationships between vocal intensity and fundamental frequency were observed over time.

Significance

Considering the vocal fold microarchitectural properties and phonation characteristics of the rabbit, the in‐vivo preparation described may provide a useful small animal option for investigations of phonation dependent tissue remodeling, maintenance of the vocal fold lamina propria, and development of pathology.

Support

NIH‐NIDCD R03 DC008400.
Cochlear Nerve Stimulation With Optical Radiation
Authors: Richter Claus‐Peter , Fishman Andrew J , Izzo Agnella D
Published: August 2008

Problem

Neural prosthetic devices are artificial extensions to the body that restore or supplement nervous system function that was lost during disease or injury. The devices stimulate remaining neural tissue with electric current, providing some input to the nervous system. Hereby, the challenge for neural prostheses is to stimulate remaining neurons selectively. However, electrical current spread does not easily allow stimulation of small neuron populations. In neural prostheses developments, particular success has been realized in the cochlear prostheses development. The devices bypass damaged hair cells in the auditory system by direct electrical stimulation of the auditory nerve. Stimulating discrete spiral ganglion cell populations in cochlear implant users’ ears is similar to the encoding of small acoustic frequency bands in a normal‐hearing person's ear. In contemporary cochlear implants, however, the injected electric current is spread widely along the scala tympani and across turns. Consequently, stimulation of spatially discrete spiral ganglion cell populations is difficult.

Methods

Spiral ganglion cells in guinea pigs were stimulated with laser pulses from an Aculight Capella infrared laser.

Results

With our experiments we demonstrate that extreme spatially selective stimulation is possible using light.

Conclusion

Our long‐term goal is to develop and build an optical cochlear implant prosthesis to stimulate small populations of spiral ganglion cells.

Significance

Our long‐term goal is to develop and build an optical cochlear implant prosthesis to stimulate small populations of spiral ganglion cells.

Support

This project has been funded with federal funds from the National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Department of Health and Human Services, under Contract No. HHSN260‐2006‐00006‐C / NIH No. N01‐DC‐6‐0.
Cortical Processing of Pitch: Excitation or Inhibition?
Authors: Harris Robert L , Nagina Subrati , Jones Steve John
Published: August 2008

Problem

Although tonotopicity provides frequency representation, it does not provide an explicit representation of pitch for complex sounds. For conventional stimuli, it is not possible to vary pitch without varying the spectrum of the sound. It is possible using a regular interval noise(RIN). RIN isolates the neural response associated with the perception of pitch. Several recent studies have identified a specific region in the auditory cortex that contains a cortical representation of pitch.

Methods

RIN auditory evoked potentials to: 1) determine over what range of temporal periods of RIN evoked a pitch onset response (POR); 2) determine how many cycles of RIN are required for its detection; 3) investigate inter‐hemispheric predominance of the POR; 4) investigate if different periodicities are represented in different cortical regions; 5) determine if the degree of refractoriness of the POR is similar when the same pitches occur at regular, short intervals, as when infrequent occurrences of a particular pitch are interspersed with other pitches at the same short intervals.

Results

1) a pitch onset response is evoked over the whole range of periodicities giving rise to a sense of musical pitch; 2) at most periodicities less than 2 cycles of RIN are required in order to evoke a response; 3) no marked interhemispheric differences were revealed; 4) evidence for a “periodotopic” distribution of responses was inconclusive; 5) the refractory properties of the response mainly suggested a common neuronal population, but with small degree of pitch specificity.

Conclusion

There is a striking apparent disparity between human studies, which suggest a large increase in both metabolic and electrophysiological activity when random noise becomes periodic, at whatever pitch, and the animal literature which suggests only a few pitch‐selective neurons are excited. We propose a model that explains the observed differences: that the large neuronal population response represents widespread inhibition.

Significance

The investigation of patients with auditory processing disorders.