bluuebunny/crossref_metadata_2025 huggingface dataset:
Inflammation and Sudden Hearing Loss
Authors: Kassner Stefan S , Stern‐Straeter Jens , Bran Gregor , Hoermann Karl , Kinscherf Ralf , Goessler Ulrich R.
Published: August 2008

Problem

Although sudden hearing loss (SHL) remains to be a frequent indication for emergency treatment in ENT departments, the pathogenesis leading to this disease is yet unknown. There is increasing evidence that vascular risk factors like hypercholesterolemia and chronic inflammation contribute to a disease‐provoking environment and, thus, enhance the risk for sudden hearing loss. We have shown in the past that patients with acute ischemic stroke display signifantly elevated plasma levels of oxLDL as well as significantly elevated gene expression of proinflammatory (CD40, TNF‐alpha and COX‐2), proapoptotic (Caspase‐3 and PARP) and proadhesive (CD38) genes. The aim of our study was to elucidate the possible role of hypercholesterolemia, inflammation and oxidative stress in sudden hearing loss and to show possible parallels with ischemic events like stroke.

Methods

Mononuclear cells from patients with SHL and healthy controls were isolated from blood and plasma. Analysis was performed using ELISA and FACS‐analysis.

Results

We measured the levels of blood lipids, oxLDL, TNF‐alpha and CD40L in the plasma of patients with sudden hearing loss in comparison to healthy controls using ELISAs. Furthermore, we isolated mononuclear cells from the blood of patients and controls and measured the protein expression of proinflammatory (CD40, TNF‐alpha and COX‐2), proapoptotic (Caspase‐3 and PARP), oxidative stress associated (Mn‐SOD) and adhesion relevant (CD38) proteins by FACS‐analysis and ELISA.

Conclusion

We found that both plasma levels of the above mentioned markers and protein expression in mononuclear cells showed distinct changes compared to healthy controls.

Significance

We therefore postulate that a proinflammatory environment and oxidative stress might trigger the onset of sudden hearing loss and influence the clinical outcome.
Ototoxicity in Guinea Pigs: NO Levels in Cochlear Fluids
Authors: Yonamine Fernando Kaoru , Mendes Jose Arruda , Okada Daniel Mochida , Onishi Ektor T , Suemitsu Higa Elisa Mieko , Mouro Margareth Gori , Neto Jose Mendes
Published: August 2008

Problem

Ototoxicity continues to be the most common side effect of gentamicin, and its physiopathology is still unclear. Currently the main accepted theory relates the ototoxicity with over production of free radicals, like nitric oxide (NO). Objective: To analyze the NO levels in the cochlear fluids of guinea pigs exposes to gentamicin and its relation with ototoxicity and propose an experimental model to study ototoxicity and drugs that may prevent it.

Methods

Six guinea pigs were used, divided in two groups: a) Study – submitted to intraperitoneal injections of gentamicin, (135mg/kg/day) for 14days; b) Control ‐ submitted to intraperitoneal injections of distilled water, daily, for 14days; the hearing loss was evaluated with Preyer's reflex. On 14 day the guinea pigs were sacrificed and the cochlear fluids were collected for dosage of the nitric oxide through the chemiluminescence method (NOA‐TM 280).

Results

The Preyer's reflex was absent at the end of the experiment in the Study group, what did not happen in the control group. The average of the nitric oxide levels of the guinea pigs of the study group was 26.2μ and the Control group 11.26μM.

Conclusion

It was possible to measure nitric oxide levels in guinea pigs cochlear fluids. The results reinforce the role of nitric oxide in ototoxicity induced by gentamicin.

Significance

Experimental study (Level 2).
Phase I and Animal Studies of the TACA Implanted Hearing Aid
Authors: Slattery William H , Richmond Keith Neu , Kunz Lawrence Leroy , Janis Abram Daved
Published: August 2008

Problem

Hearing aid technologies are lacking in terms of cosmesis and comfort, encumber activities such as showering and swimming, and can distort the user's voice due to occlusion effect. The Transcutaneous Air Conduction Aid (TACA) has been developed as an alternative for patients with mild‐to‐moderate high‐frequency sensorineural hearing loss. This new device is designed to bring amplified sound into an open ear canal in an invisible manner. This study presents our investigation into the additional challenge of a percutaneous device at risk for marsupialization and infection at the device‐skin interface.

Methods

TACA phantoms, coated with a fibrous titanium mesh biomaterial, were tested in long‐term preclinical implantation studies. Histomorphometric comparison of mesh coated and uncoated TACA phantoms was performed in 3 rabbits at 90 and 180 days. A follow‐up study in 7 rabbits was performed to temporally characterize wound healing around the mesh coated implants from 1–13 weeks. Based on promising animal results, a phase I FDA human trial was conducted lasting 6 months.

Results

Preliminary histomorphometric studies (N=18 phantoms with mesh, 6 without mesh) in 3 rabbits demonstrated that the titanium mesh biomaterial prevented marsupialization and infection at 90 and 180 days. The follow‐up study of 42 mesh‐coated phantoms in 7 rabbits demonstrated that the epidermal seal was reestablished around the percutaneous projection at 3 weeks, with supporting vascularized connective tissue ingrowth complete at 6 weeks. The human phase I data demonstrated connective tissue ingrowth and epidermal sealing with no evidence of marsupialization or infection.

Conclusion

The TACA may offer an alternative for patients with sensorineural mild‐to‐moderate sloping high frequency hearing loss. Percutaneous implant studies in rabbits and man demonstrated the safety of this novel percutaneous device.

Significance

Preclinical and clinical demonstration of feasibility and safety.

Support

This work was supported by Implanted Acoustics, Santa Clarita, California.
Jagged 1 is Necessary for Palatal Arterial Development
Authors: Humphreys J. Ryan , Goudy Steven L
Published: August 2008

Problem

Palatal development is a complex process of growth, elongation, and fusion of paired palatal shelves. We hypothesize that arterial formation is necessary for normal palatal growth and development. Alagille's syndrome, a recently categorized genetic syndrome, is linked to genetic mutations in Jagged 1, a Notch signaling ligand necessary for vascular development. The cranial neural crest (CNC) cells are a well known cell population crucial in palate formation. To test our hypothesis, we have selectively removed Jagged 1 in the CNC population.

Methods

We identified the normal pattern of Jagged 1 expression in the palate using in situ hybridization and immunohistochemistry. We selectively removed Jagged 1 from the craniofacial mesenchyme using Cre‐Lox technology, mutant Wnt1‐cre Jagged 1 f/f selective knockouts. Mutant embryos were harvested at e13, e14, and e15 (critical time points in palatal development). To visualize the vascular development, we used platelet endothelial cell adhesion molecule (PECAM) antibody staining and utilized immunoflouresence with confocal microscopy.

Results

Jagged 1 gene and protein expression was found throughout the midface and palate. The expression of Jagged 1 was concentrated in the perivascular tissue in the palatal mesenchyme. As palatal development occurs, there is a concomitant increase in Jagged 1 expression. Wnt1‐cre Jagged1 f/f mice were found viable. However, the mutant mice died in the neonatal period secondary to severe midface hypoplasia. Mutant mice had a significantly hypoplastic osseous palate and irregularly spaced palatal rugae. Examination of e15 mutant palatal shelves, using PECAM immunoflouresence and confocal microscopy, demonstrated a complete absence of mature vasculature.

Conclusion

Jagged1 is required for palatal vascular development. The absence of Jagged1 in the CNC leads to severe palatal hypoplasia and neonatal death.

Significance

Cleft lip and palate is the most common craniofacial congenital deformity occurring in 1:1000 live births. Clefting in conjunction with vascular syndromes, such as Alagille's, is a common observation.
Effectiveness of Honey on S.aureus and P.aeruginosa Biofilms
Authors: Alandejani Talal , Marsan Joseph G , Ferris Wendy , Robert Slinger , Chan Frank
Published: August 2008

Problem

Biofilms formed by Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA) have been shown to be an important factor in the pathophysiology of chronic rhinosinusitis (CRS). Growth in biofilms increases bacterial resistance to antibiotics, which may explain why CRS responds poorly to antibiotic therapy. Honey has been used as an effective topical antimicrobial agent in the treatment of antibiotic‐resistant bacterial wound infections. To our knowledge, no research has studied the effect of honey on biofilms.

Methods

We used a previously established biofilm model to assess antibacterial activity of Manuka honey from New Zealand and Sidr honey from Yemen. These were tested at a 1 in 2 dilution against 11 isolates of methicillin‐susceptible SA (MSSA), 11 methicillin‐resistant SA (MRSA), and 11 PA isolates. Honeys were tested against both planktonic (broth) and biofilm‐grown bacteria.

Results

Both honeys were effective in killing 100% of the isolates in the planktonic form. Biofilm‐grown bacteria were less susceptible to the honeys, but honey still had significant bactericidal activity. The bactericidal rate for the Sidr and Manuka honeys against MSSA MRSA and PA were 63%, 73%, 91% and 82%, 63% and 91%, respectively. These rates were significantly higher (p<0.001) than those seen with single antibiotics commonly used against MSSA and MRSA (e.g. cloxacillin and vancomycin) in a previous study done on the same bacterial isolates.

Conclusion

Honey, which is a natural, non‐toxic and inexpensive product, is effective in killing SA and PA bacterial biofilms.

Significance

This intriguing observation may have important clinical implications and could lead to a new approach for treating refractory CRS.
Electrophysiologic Features of Freshly Excised Nasal Tissue
Authors: Cho Do‐Yeon , Beate Illek , Horst Fischer , Hwang Peter H
Published: August 2008

Problem

Epithelial ion transport regulates hydration of respiratory mucosal surfaces, which promotes effective mucociliary clearance. Activators of chloride ion (Cl‐) secretion, such as ascorbic acid (Vitamin C), may enhance the rheologic properties of mucus. Altered ion transport could play a role in the pathogenesis of chronic rhinosinusitis (CRS). The purpose of this study is to assess the electrophysiologic characteristics and role of vitamin C on the nasal mucosa of CRS patients.

Methods

Nasal tissues (uncinate process, inferior turbinate, nasal septum) were obtained from five CRS patients during sinus surgery and mounted on disks with open areas of 0.03cm2 to 0.71cm2 between Ussing hemichambers. Short‐circuit current (Isc) was continuously recorded, and at 50‐s intervals transepithelial voltage was clamped from 0 to 2mV. Serosa‐to‐mucosa‐directed Cl‐ gradient was applied to increase the electrochemical driving force for Cl‐ exit across the apical membrane.

Results

Isc decreased when the epithelial Na+ channel blocker (amiloride) was added to the luminal side of the chamber indicating that the tissues were Na+ absorbing. Addition of the cAMP‐elevating agonist forskolin induced a Cl‐ secretory response and exposure of the apical airway surface to vitamin C(600uM) stimulated the transepithelial Cl‐ secretion to 60% of the forskolin‐stimulated Isc. The contribution of the Na+/K+/2Cl‐ cotransporter to the Cl‐ secretory response was verified by addition of bumetamide. Glybenclamide was used to probe for the CFTR‐Cl‐ conductance. These results were observed in all specimens.

Conclusion

Freshly excised human nasal epithelium is easily accessible and its bioelectric measurements can be applied as a functional measurement of ion transport in epithelial diseases. Vitamin C may serve as a biological regulator of CFTR‐mediated Cl‐ secretion in human nasal epithelia.

Significance

Vitamin C in human nasal epithelia may represent a potential target for the complementary treatment of thickened mucus secretions by enhancing epithelial fluid secretion in diseases, such as CRS or cystic fibrosis.
In Vitro Study of Interleukin‐5,‐13,VCAM‐1 in CRS
Authors: Sauter Alexander , Stern‐Straeter Jens , Hörmann Karl , Naim Ramin
Published: August 2008

Problem

The pathogenesis of eosinophilic chronic rhino‐sinusitis (ECRS) is still unclear. Paranasal mucosa inflammation might be related to eosinophil infiltration. Eosinophil infiltration seems to induce changes in the expression of cell adhesion molecules, such as VCAM‐1. Futhermore Mediators such as Interleukin 5 and 13 (IL‐5, IL‐13) are considered to be key factors for eosinophilic accumulation and activation. The E ‐ Cadherin – β ‐ catenin complex maintain the integrity of epithelium. Down‐regulation of β‐catenin and E‐cadherin is a pivotal factor for loose integrity and progressive cell growth. This study aimed to assess which cytokines regulate the expression of the adhesion molecule E‐cadherin and the multifunctional protein β‐catenin, playing an essential role in the cadherin‐mediated anchoring, in ECRS.

Methods

ECRS cell cultures were incubated with IL‐5, IL‐13 and VCAM‐1 and β‐catenin / E‐cadherin levels were analysed after 8–72 hours using cytokine immunoassay and immuno‐histochemistry.

Results

We could ascertain a significant increase in β‐catenin expression in eosinophilic paranasal cell culture after IL‐13 administration compared to non‐eosinophilic culture. Stimulation with IL‐5 did demonstrate an alteration of E‐cadherin expression. Incubation with VCAM‐1 could induce a significant increase in E‐cadherin expression in fibroblast cell cultures. Immuno‐staining for β‐catenin was restricted to the membrane of the cells.

Conclusion

In regard to the increased mural expression of β‐catenin, we presume that a fibrotic reaction similar to asthma and chronic obstructive pulmonary disease is taking place in patients suffering from CRS.

Significance

The tissue remodelling during chronic eosinophilic inflammation offers new insight into pathogenesis of ECRS.
Safety of Ethmoid Sinus Drug‐Eluting Catheter Insertion
Authors: Melroy Christopher T , Kuhn Frederick A
Published: August 2008

Problem

The objective is to develop an instrument which allows a drug‐eluting catheter to be safely and reproducibly inserted into the ethmoid sinuses.

Methods

A trochar‐based insertion device was designed to allow delivery of a drug‐eluting catheter into the anterior and posterior ethmoid sinuses. It was inserted into 12 cadaveric ethmoid sinuses under endoscopic and fluoroscopic guidance. CT scans were performed pre‐, intra‐, and post‐procedure. The device's position was analyzed and the proximity to the skull base, lamina papyracea, and ethmoid face was measured. The specimens were then dissected and evaluated for skull base, sphenoid face, or lamina papyracea injury.

Results

The drug eluting catheter system was successfully inserted into the ethmoid sinuses of all 12 cadaver sides without injury to either the medial orbital wall or the skull base as confirmed by post‐procedure CT scan and dissection. The final position of the distal tip of the stent averaged 8.1mm (RMS = 3.3) from the skull base, 5.6mm (RMS=3.5) from the sphenoid face, and 5.0mm (RMS=3.5) from the lamina papyracea; the proximal tip was at the face of the ethmoid bulla and 17.1mm (RMS=3.5) below the skull base.

Conclusion

This study demonstrates that a trochar‐based instrument can safely and reproducibly introduce a drug‐eluting catheter into the ethmoid sinuses without skull base or lamina papyracea injury. This device may allow safe topical drug delivery into the ethmoid sinuses and provide chronic ethmoid sinusitis patients an alternative to ethmoidectomy.

Significance

The primary surgical therapy for chronic ethmoid sinusitis is ethmoidectomy; topical therapy has been widely used in the management of chronic ethmoid sinusitis only after ethmoidectomy. This study shows a drug‐eluting catheter can be safely and reliably inserted into virgin ethmoid sinuses in order to allow the topical elution of medications into the ethmoids without ethmoidectomy.

Support

Acclarent supplied cadaveric specimens.
The Impact of Olfactory Stimulation on Dreams
Authors: Stuck Boris A , Desislava Atanasova , Grupp Kathrin Frauke , Schredl Michael
Published: August 2008

Problem

Only a limited number of trials is available regarding the impact of external and internal stimuli on human dreams. Current research focuses on the question whether these stimuli (acoustic or mildly painful stimuli e.g.) are incorporated into dreams and whether they influence their emotional coloring. In a recently published trial we were able to demonstrate that isolated olfactory stimulation does not lead to arousals. Aim of the present study was to investigate whether olfactory stimuli of different hedonic characteristics influence dreams in humans.

Methods

15 young healthy female volunteers were investigated during 30 nights of testing. Standardized awakenings were performed during REM sleep. During REM phases, subjects were exposed to non‐odorous control, a positive odor (PEA, 20%) and a negative odor (H2S, 4 ppm) for 10 seconds each in a randomized fashion using a computer olfactometer. After the awakening, subjects were advised to report the content of their dream in a standardized fashion and to assess the emotional coloration (positive and negative) on a four digit scale (0/1/2/3: no / little / moderate / strong feelings) to calculate the overall emotional coloration.

Results

In all subjects, three REM awakenings were performed. For 40 out of 45 awakenings, a dream was reported by the subjects. Mean emotional coloration after control stimulation was slightly positive (+0.5). After negative stimulation, the mean emotional coloration was shifted to negative values (‐0.4) while the mean emotional coloration was significantly more positive after positive stimulation (+1.2).

Conclusion

Standardized REM awakenings can be performed successfully even under the conditions of an olfactory laboratory with intransal stimulation. With olfactory stimulation, the emotional coloration of dreams can be significantly influenced in accordance with the hedonic aspect of the stimulant.

Significance

The study opens a potential field of therapeutic intervention with nocturnal olfactory stimulation.
R400 – Effects of Silicone Gel Sheeting on Scar Formation
Authors: Aminpour Shervin , Tollefson Travis T
Published: August 2008

Problem

The treatment of keloids and hypertrophic scars has long been a concern for facial plastic surgeons. There are a wide variety of over‐the‐counter products that claim to improve hypertrophic scarring. Post‐surgical patients often inquire as to which of these products will give the best cosmetic result. Silicone gel sheeting is reported to provide positive outcomes with respect to a reduction in scar hypertrophy and an improvement in color differences. The exact mechanism is unknown. Micropore (3M) tape, a cheaper and more readily available product, has also been reported to reduce scar hypertrophy. This study will evaluate the differences and effectiveness of silicone gel sheeting and micropore tape in the reduction of scar hypertrophy in an animal model of scarring.

Methods

Hypertrophic scar formation was induced in an established rabbit model that parallels human skin. Treatment of the animals' scars was divided into 3 study groups: silicone gel sheeting, micropore tape, or no treatment. 80 total scars were treated for 30 days after the complete reepithelialization of the created scars. Both photometric and histologic analysis was performed on the scars. Scar hypertrophy was analyzed with blinded observers using a visual analog scale. Histological analysis was performed using the Scar Elevation Index, a ratio of the scar height over normal skin.

Results

Both photographic and histological analysis showed there was a statistically significant improvement (p<0.05) in scar hypertrophy after use of either silicone gel sheeting and micropore tape when compared with controls.

Conclusion

Silicone Gel Sheeting and Micropore Tape are equally effective in the reduction of hypertrophic scarring.

Significance

Facial plastic surgeons should consider covering facial incisions with micropore tape for 30 days after surgery to minimize the risk of hypertrophic scarring.
R401 – Anti‐Aging Compounds and Epidermal Effects on Mouse Skin
Authors: Bhattacharyya Tapan K , Higgins Natalie P , Sebastian J Scott , Thomas J Regan
Published: August 2008

Problem

To compare topical effects of 5 anti‐aging agents on epidermal histology in the non‐irradiated hairless mouse.

Methods

Female retired breeders (SKH‐1 hairless mouse) were treated topically on the dorsal skin with 5 commercially available agents as a daily treatment for 2 weeks. The agents used were retinoic acid, glycolic acid, estrogen, soy, and vitamin C. Skin sections were analyzed in the light microscope to acquire morphometric data of keratinocyte proliferation (Index of Proliferating Cell Nuclear Antigen), epidermal thickness, and nuclear volume of cells from three epidermal layers. Quantitative data were analyzed with SPSS 14.0 statistical software to detect significant difference between the means of control and 5 experimental groups.

Results

Epidermal stimulation was observed with all cosmeceuticals but most pronounced effects resulted from the application of glycolic acid, and estrogen, and retinoic acid to a somewhat lesser degree. Soy and vitamin C produced significant cellular changes of lesser magnitude.

Conclusion

Reported studies of many of these agents were conducted in the irradiated mice, and their effects on the intact animal skin have been rarely documented. These data will be useful for our contemplated studies of photo aging in the same species. Agents like retinoic acid derivatives, or glycolic acid are popular rejuvenating compounds, but have been reported to produce annoying side effects. Milder compounds like Vitamin C or soy may cause reversal of detrimental aging changes without cutaneous side effects.

Significance

This study may be helpful to pinpoint therapeutic advantages of popular anti‐wrinkle compounds, and devise clinical strategies to combat photo aging.

Support

This research has been supported by the 2007 Leslie Bernstein Grant from the AAFPRS Foundation.
R402 – Profilometry and Skin Histology with Anti‐wrinkle Agents
Authors: Bhattacharyya Tapan K , Linton Jeannie , Mei Lily , Thomas J Regan
Published: August 2008

Problem

To investigate whether topical anti‐aging compounds can reduce wrinkle depth as seen by replica profilometry with comparable changes in histological parameters in murine skin.

Methods

Female hairless mice were exposed to topical application of a retinoic acid cream, a peptide lotion, and a soy cream on the dorsal skin areas for 4 weeks. Silicone negative replicas of treated and untreated skin surface were photographed using Optimus V6.5 image processing system (CuDerm Corporation, Texas), and evaluated for traditional surface roughness parameters (Rz, Ra, FSpace, and DL). Skin samples were also processed for histomorphometry, and immunohistochemistry of proliferating cell antigen (PCNA). Quantitative light microscopic data were acquired from tissue sections for estimating DNA replication in epidermal keratinocytes (PCNA index), epidermal thickness, depth of dermal collagen bundles.

Results

All quantitative data were analyzed for comparing means with one‐way analysis of variance (ANOVA) using SPSS 14.0 statistical software, and significant changes in all the parameters were noted. Augmented keratinocyte proliferation and thickening of viable epidermis was observed with all three compounds although a greater effect was seen in the retinoic acid and peptide treatment groups. A similar trend was noted in respect of widening of dermal collagen layer. Epidermal surface roughness manifested maximum smoothening effect following treatment with the peptide compound.

Conclusion

Retinoic acid effects have been reported in literature but reports of comparison with soy and peptide preparation in this laboratory model are lacking. The pronounced effects seen with all the compounds indicate topical agents other than retinoic acid may have beneficial regenerating effects on photo aged skin without irritating effects reported for retinoic acid.

Significance

These results may be helpful in defining clinical strategies for topical non‐invasive improvement of aging facial skin.

Support

This study was supported by the 2007 Leslie Bernstein Grant from the AAFPRS Foundation.
R403 – Epidermal Proliferating Cell Nuclear Antigen in Aging Rats
Authors: Bhattacharyya Tapan K , Jackson Paula , Thomas J Regan
Published: August 2008

Problem

To determine if epidermal cell proliferation in colony‐raised Fischer 344 rats changes with age and diet.

Methods

Fischer 344 rats fed ad libitum and calorie‐restricted (CR) diets were obtained from the NIA colonies, and young, young adult, and old animals from both groups were used for this study (six in each group). Tissue sections from the dorsal skin (DS) and foot plate (FP) were used for immunohistochemical staining of proliferating cell nuclear antigen (PCNA). The proliferation index (PCNA‐I) was computed from counts of stained and total number of keratinocytes. Simultaneous measurements of epidermal thickness were obtained from same sections. Data were analyzed with Excel and SPSS 14.0 software for statistics.

Results

Two‐way analysis of variance (ANOVA) was applied to the data to probe the effect of age, diet, and age‐diet interaction. A significant effect of age was noticed in the two parameters i.e., DS PCNA (F 3.96, P. 011), FP epidermal width (F 3.37, P. 021) and FP PCNA‐I (F 9.0, P. 000). A significant correlation between DS width and PCNA values was also noted (r 0.5, P. 01).

Conclusion

There is a trend of reduction of PCNA positive cells with increasing age irrespective of thickness of epidermis, and this trend is more apparent in CR rats.

Significance

This cell proliferation study has implications in relation to CR effect on age‐related disease conditions, and biogerontology.

Support

The study was partially funded by the 2007 Leslie Bernstein grant from AAFPRS foundation.
R404 – Regulatory T Cells and Clinical Outcome in HNSCC Patients
Authors: Alhamarneh Osama , Stafford Nicholas D. , Greenman John
Published: August 2008

Problem

To determine the correlation between peripheral blood CD4+CD25high regulatory T cells (Treg), a suppressor cell population that dampen the immune response, and clinical outcome and survival in HNSCC patients.

Methods

Treg cell numbers in the peripheral blood of newly‐presenting, untreated HNSCC patients (n=65) were determined pre‐operatively, 4–6 weeks after treatment (n=30) and in a cohort of healthy controls (n=35) of similar age and sex, after Treg cell isolation using magnetic microbeads (Miltenyi Biotec) by flow cytometry. The Mann‐Whitney U test was used to analyse the correlations between Treg cell levels and clinical outcome.

Results

Treg cells were significantly higher in patients pre‐operatively vs. controls (p=0.002). After treatment, patients showed a significant rise compared with their pre‐treatment levels (p=0.022). Pre‐treatment Treg cells levels did not correlate with survival or any of the other conventional clinicopathological parameters. However, higher Treg cells levels were discovered in the advanced disease stages (III/IV vs. I/II, median 6.3 vs 4.3) in the pre‐treatment group that turned into significantly higher levels in the early disease stages post treatment (I/II vs. III/IV, median 10.8 vs. 5.67 p=0.044).

Conclusion

Although peripheral blood Treg cells levels were higher in patients when compared to controls, no correlation was found between this cell population and clinical outcome or survival. In contrast with gastric, colorectal and ovarian tumors, Treg cell levels did not normalize 4–6 weeks after curative treatment in this cohort of HNSCC patients. Studies into Treg cell function are thus required to try and elucidate the apparent paradox in Treg cell levels observed in HNSCC.

Significance

The presence of Regulatory T cells in the peripheral blood of HNSCC patients may be detrimental to host defence against tumor. Further studies are needed to explore their role in the tumor microenvironment and their correlation with clinical outcome.
R405 – Laser Welded Transluminal Repair of Rabbit Esophageal Injury
Authors: Bleier Benjamin Saul , Gratton Michael Anne , Leibowitz Jason M , Palmer James N , Newman Jason G , Cohen Noam A
Published: August 2008

Problem

To test the feasibility of Laser Tissue Welding in creating an endoscopic transluminal repair of esophageal perforation.

Methods

A diode laser was used to create an endoluminal rabbit esophageal perforation repair. Burst pressures were compared to open incision, external suture, and external laser augmented suture closure. Comparisons were performed 5 times and analyzed using Kruskal‐Wallis ANOVA and a post‐hoc Dunn's Method.

Results

The burst threshold of the endoluminal weld (54.78mHg) was significantly higher than the open incision (6.5mmHg) and not significantly different than the external suture (37.18mmHg) or the laser augmented suture group (71.60mmHg).

Conclusion

Laser welding is a feasible method of creating endoluminal repairs with burst strengths comparable to external suture repair.

Significance

This is the first report of Laser Tissue Welding for endoscopic closure of iatrogenic esophageal perforation which may allow a subset of these patients to avoid traditional open approaches.
R408 – Management of the Parathyroid Carcinoma
Authors: Gérald Valette , Potard Gaël , Martins‐Carvalho Christine , Gac Marie‐Suzanne Le , Marianowski Remi
Published: August 2008

Problem

Management of parathyroid carcinoma.

Methods

2 cases of parathyroid carcinoma managed in Brest Hospital, Ear, Nose and Throat Service (France).

Results

Through a review of the literature we discuss the initial surgical treatment for this rare tumor, which has a strong potential of local recurrence or metastasis (fifty percent).

Conclusion

The treatment of the parathyroid carcinoma is surgical. It requires a total ablation of the parathyroid tumor, of the homolateral thyroid lobe, of the homolateral recurrent nodular tissues and of sticking tissues.

Significance

A large ablation is required.
R413 – KITENIN Enhances Invasion and Metastasis in FOM Cancer Model
Authors: Lee Joon Kyoo
Published: August 2008

Problem

KAI1 is a metastatic suppressor gene. The expression of KAI1 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). KITENIN is reported to promote metastasis in a mouse colon cancer model. The purpose of this study was to investigate tumor invasiveness and early lung metastasis by KITENIN in murine floor of mouth cancer model.

Methods

The cDNA of KITENIN and the vector only were transfected into the SCC VII (mouse squamous cell carcinoma cell line) cells. The transfections were performed using the FuGENE 6 transfection reagent. The suspension of 5 × 105/? viable KITENIN‐ or vector‐transfected SCC VII cells were injected into the floor of mouth of C3H/HeJ syngeneic mice deep to the mylohyoid muscle via an intra‐oral approach. From the 1st week to 6th week after injection, tumor, lung, liver, and brain tissues were obtained from six mice each group every week and were evaluated under the light microscope.

Results

On each group, the tumor invaded to superficial musculature of floor of mouth, deep musculature of floor of mouth, salivary gland, perineural tissue, bone, and skin sequentially. Lung metastases started from peripheral and localized region and moved to central and diffuse lung tissues. Earlier and more aggressive tumor invasion to deep floor of mouth musculature, salivary gland, perineural tissue, bone, and skin was observed in KITENIN‐transfected group than in vector group. Earlier and more diffuse lung metastasis was observed in KITENIN group.

Conclusion

KITENIN enhanced tumor invasion and lung metastasis in murine floor of mouth squamous carcinoma model.

Significance

An antisense KITENIN strategy could be used to inhibit distant metastasis and progression in head and neck squamous cell carcinoma.

Support

This study was supported by a grant CUHRICM‐CRI07038‐1.
R416 – Bipolar Cautery: Sunderland Third Degree Nerve Injury Model
Authors: Moradzadeh Arash , Brenner Michael Joel , Whitlock Elizabeth , Luciano Janina Paula Delpilar , Myckatyn Terence Michael , Daniel Hunter , Tong Alice , Mackinnon Susan E.
Published: August 2008

Problem

The prognosis of bipolar electrocautery nerve injury is unpredictable because few experiments compare it to characterized nerve injury models.

Methods

16 Lewis rats were randomized to sciatic crush or bipolar cautery and at 21 days evaluated with nerve morphometry and walking tack analysis. Double transgenic Thy1‐CFP/S100‐GFP mice were used to serially image axonal regeneration and Schwann cells (SC) over time following injury.

Results

In rats, bipolar cautery injury shows greater disruption of myelin and neurofilament architecture at the injury site. There is decreased total nerve fiber counts distal to the injury (p<0.05). Walking track analysis demonstrates functional recovery after crush, but not after cautery injury. Serial imaging of mice, shows axonal regeneration starting at week 1 after crush, but late, partial axonal regeneration in the cautery group; these findings were reflected in endplate reinnervation at 42 days.

Conclusion

Results suggest that a Sunderland type 3 injury, characterized by slow, variable, incomplete recovery, results from a bipolar electrocautery injury.

Significance

Bipolar cautery injuries can be observed without immediate surgical intervention.
R418 – Temsirolimus Preferentially Targets Tumor Microenvironment
Authors: Nathan Cherie Ann O , Ekshyyan Oleksandr , Dennis D'Antoni Carmichael , Clark Cheryl , Youhua Rong , Xiaohua Rong , Abreo Fleurette
Published: August 2008

Problem

The mTOR pathway, a central regulator of cell metabolism, proliferation, and survival, is activated in a 100% of HNSCC. We have previously shown the mTOR inhibitor CCI‐779 (temsirolimus; rapamycin analogue) inhibits growth of FaDu HNSCC in vitro and in vivo and significantly reduced microvessel density in HNSCC xenografts. Hence to study stromal effects of temsirolimus, FaDu cells were stably transfected with rapamycin resistant mTOR (rr‐mTOR) to elucidate anti‐tumor mechanisms of temsirolimus.

Methods

FaDu cells stably transfected with control vector or rr‐mTOR. HNSCC tumor xenografts were established in nude mice by subcutaneous injection of control vector transfected (cv‐FaDu) or rr‐mTOR transfected (rr‐FaDu) cells. Temsirolimus (5 mg/kg) was evaluated in mice treated for 3 weeks. PCR analysis of the 5′‐end of an exogenous rr‐mTOR sequence was performed on DNA isolated from explanted tumor tissues.

Results

Growth of cv‐FaDu was suppressed by rapamycin (100 ng/ml), while the rr‐FaDu clone was insensitive. PCR assay yielded the expected 246 b.p. product in rr‐FaDu, but not in cv‐FaDu xenografts indicating the presence of the exogenous rr‐mTOR coding sequence in rr‐FaDu xenograft tumor tissues. Temsirolimus inhibited growth of cv‐FaDu and rr‐FaDu xenograft tumors and was comparable in both groups suggesting the ‘anti‐tumor’ activity of the drug in vivo is predominantly a consequence of its anti‐stromal effects.

Conclusion

Results of this study suggest tumor growth inhibitory effects of temsirolimus on HNSCC xenografts are mediated mainly by the anti‐stromal activity of the drug rather than by direct anti‐tumor effect alone.

Significance

Understanding the mechanism of action of mTOR inhibition is important in the management of HNSCC as the Akt/mTOR pathway is activated in 100% of HNSCC.

Support

This work was supported by NCI grant R01CA102363 to Cherie‐Ann O. Nathan.
R419 – Surgery with Molecular Fluorescence Imaging Guidance
Authors: Nguyen Quyen To , Olson Emilia , Aguilera Todd A. , Vorasubin Nopawan , Jiang Tao , Tsien Roger Y.
Published: August 2008

Problem

Currently, surgical excision of tumors is dependent on the surgeon's ability to differentiate tumor from normal tissue using non‐quantifiable criteria such as tumor location, texture, color, relationship to surrounding structures etc. Under these circumstances, the ability to detect/treat micro‐metastases and residual tumors in real time is limited.

Methods

We have developed a more objective means of visualizing tumor markers utilizing a novel strategy for selectively delivering molecules to cancer cells using activatable cell‐penetrating peptides (ACPPs), in which the fluorescently labeled cell‐penetrating peptide (CPP) is coupled via a cleavable linker to a peptide that inhibits the uptake of the CPP. Upon exposure to proteases secreted by tumor, especially matrix‐metalloproteinases, the linker is cleaved, dissociating the inhibitory peptide and allowing the CPP to bind to and enter tumor cells.

Results

We injected Cy5‐labeled ACPP into transgenic mice with spontaneous tumors as well as mice xenografted with human cancer cell lines (including sarcoma and carcinoma derived from larynx, breast and prostate). All tumors retain the Cy5 label to a greater extent than normal tissues. Furthermore, xenografts derived from cancer cell lines that were stably transfected with GFP or RFP show colocalization of the FP marker with the Cy5‐labeled ACPPs. Finally, in the same mouse models, tumors excised using ACPP guided molecular imaging showed improved completeness of tumor removal as quantified by Alu PCR.

Conclusion

We have developed a novel method for molecular imaging of tumors in vivo using ACPPs. Using this technique, we were able to improve completeness of tumor excision in murine models.

Significance

This technique will help to improve complete resection of tumor at the time of surgery, thereby reducing patient morbidity in terms of total time under anesthesia and residual tumor as well as reducing operating room costs.

Support

K08 EB008122 from NIH‐NIBIB to Quyen Nguyen. Era of Hope Innovator Award from Department of Defense to Roger Tsien.
R420 – Radiation Induces TGF‐β and Blocks MyoD Causing Fibrosis
Authors: Peng Grace Lee , Masood Rizwan , Sinha Uttam K
Published: August 2008

Problem

One of the main treatment modalities for head and neck cancer (HNC) is post‐operative radiation therapy (RT). Patients undergoing RT develop dysphagia due not only to salivary gland damage, but also scarring of the pharyngeal and strap musculature. We have compared the levels of TGF‐β in the strap muscle samples in patients at the beginning and end of their RT and found that TGF‐β may contribute to excessive fibrosis by blocking MyoD expression, a gene necessary in muscle differentiation and repair. A second part of this study views the effects of concurrent transcutaneous neuromuscular electrical stimulation (NMES) during RT on TGF‐β and MyoD levels. Our objective is to examine not only the effects muscle repair post‐radiation, but also to view the effects of electrical stimulation during concurrent radiation therapy on the expression of TGF‐β and MyoD.

Methods

We used muscle samples from patients of various stages of HNC pre‐radiation and post‐radiation, with some receiving NMES during RT. The TGF‐β and MyoD expression were measured using western blotting and immunohistochemistry staining.

Results

TGF‐β expression was higher in all samples post RT as compared to non‐radiated controls. In addition, there is decreased fibrosis seen in patients who have undergone RT and concurrent NMES as compared to patients who underwent RT alone. We also observed that those patient specimens with higher TGF‐β showed lower expression of MyoD than patients with lower TGF‐β expression.

Conclusion

RT induced higher TGF‐β which blocked MyoD expression and muscle differentiation, inducing fibrosis. We hope to study different techniques of electrical stimulation during patients' RT in order to optimize the stimulation.

Significance

Understanding the mechanism through fibrosis develops can help us to increase the quality of life of patients who successfully undergo cancer research and eradication with radiation therapy. Practicing transcutaneous NMES can help to decrease the dysphagia seen in patients using RT.
R421 – Ubc9: A Novel Marker of Lymph Nodes Metastases in SCCHN
Authors: Ronen Ohad , Malone James P , Ran Sophia
Published: August 2008

Problem

Lymph node metastases (LNM) are the single most significant prognostic factor in squamous cell cancer of the head and neck (SCCHN). Therapy often relies on the ability to detect disease in cervical lymph nodes. We previously demonstrated that Ubiquitin‐conjugating enzyme (Ubc9), an enzyme involved in post‐translational modification, is over‐expressed in SCCHN compared to peritumoral adjacent mucosa. Furthermore, Ubc9 is not expressed in normal mucosa. The present study was designed to examine the expression of Ubc9 in SCCHN LNM and to determine whether Ubc9 may serve as a marker.

Methods

A portion of primary tumor, normal adjacent mucosa, when available, and metastatic lymph nodes were harvested from patients with a histologic diagnosis of SCCHN. Tissue specimens were immediately snap‐frozen in liquid nitrogen. The frozen tissue specimens were stained with antibodies to Ubc9 using standard Immunohistochemical techniques.

Results

Twelve patients with SCCHN LNM were available for staining. Sites of primary tumor specimens included larynx (n=7) and oropharynx (n=3). All of the lymph nodes had strong expression of Ubc9 in the metastatic deposits of SCCHN but not in the surrounding lymph node tissue.

Conclusion

Detection of Ubc9 expression in cervical lymph nodes from patients with SCCHN may serve as a distinct molecular marker for the presence of lymph node metastases. The findings also suggest that Ubc9 plays an important role in tumorigenesis and possibly tumor progression of head and neck squamous cell cancer. Further exploration of Ubc9 in head and neck cancer is warranted.

Significance

Ubc9 expression in SCCHN LNM can increase the sensitivity of diagnosis in the primary site as well as in suspected lymph node metastases. Early and accurate diagnosis of LNM can potentially improve regional control of the disease.

Support

Part of the study was supported by a grant from SIU School of Medicine.
R425 – Association of CD44 v6 and v10 Isoforms In HNSCC Progression
Authors: Wang Steven J , De Heer Anne‐Martine
Published: August 2008

Problem

The CD44 family of receptors includes multiple variant isoforms, several of which have been linked to tumor progression. The objective of this study was to investigate the role of CD44 v6‐ and v10‐containing isoforms in head and neck squamous cell carcinoma (HNSCC) progression.

Methods

A laboratory investigation utilizing a panel of 13 HNSCC cell lines and clinical tissue specimens from a series of 82 patients with HNSCC was performed.

Results

Immunohistochemical analysis of 82 primary HNSCC tumors and 24 metastatic lymph nodes revealed expression of CD44 v6 and v10 variants. A greater proportion of metastatic lymph nodes vs. primary tumors demonstrated strong expression of CD44 v6 (18/24 vs. 26/82) and CD44 v10 (14/24 vs. 16/82), while expression of standard CD44 was not significantly different in metastatic lymph nodes and primary tumors (10/24 vs 60/82). In addition, greater than expected expression of CD44 v10 was found among 14 specimens obtained from patients previously treated with chemoradiation. To further analyze the role of these CD44 variants in HNSCC progression, HNSCC cell lines were selected on the basis of their CD44 v6 and v10 expression. A HNSCC cell line without CD44 expression and cells of the same variant‐expressing cell lines transfected with short‐hairpin RNA for CD44 were used as controls. Compared to controls, HNSCC cells expressing CD44 v6‐containing isoforms demonstrated increased hyaluronan‐mediated tumor cell growth and migration. Furthermore, HNSCC cells expressing CD44 v10‐containing isoforms demonstrated increased hyaluronan‐mediated cisplatin resistance.

Conclusion

CD44 v6‐ and v10‐containing isoforms are associated with HNSCC progression.

Significance

CD44 v6‐ and v10‐containing isoforms may be useful tumor markers and targets for HNSCC therapy.

Support

This work was supported by an AAO‐HNS/AHNS Young Investigator Award.
R427 – Mechanism Producing Subglottic Air Pressure While Swallowing
Authors: Gross Roxanne D , Carrau Ricardo L , William Slivka , Smith Libby J
Published: August 2008

Problem

The importance of positive subglottic air pressure during swallowing has been recognized, yet no experiment has sought to determine the mechanism that generates this pressure. We hypothesized that transpulmonary recoil forces are responsible for this pressure. Objective: To compare direct subglottic air pressure measurements obtained while swallowing at different lung volumes with airway pressures obtained by total respiratory system recoil measurements.

Methods

Direct measurement of subglottic air pressure during swallowing was obtained via a percutaneous puncture of the cricothyroid membrane. Tidal volumes were measured via a heated pneumotachometer as the subjects breathed through a nasal mask. Next, to model swallowing, airway pressures were measured at the mouth by occluding the airflow for a duration consistent with true vocal fold closure during swallowing. Passive exhalation was used to mimic the central inhibition of respiratory muscles that occurs during the swallow.

Results

Regression analyses on the combined data showed strong linear relationships between lung volumes (X) and airway pressures (Y) with overall goodness of fit which were highly significant (p values less than 0.00001). R Square values ranged from 0.78 to 0.91.

Conclusion

The subglottic air pressure during a swallow is likely generated by transpulmonary recoil forces that dominate when respiratory muscles are inhibited. The degree of subglottic pressure produced is directly related to lung volume.

Significance

The influence of the respiratory phase, lung volume, and available transpulmonary recoil pressure may alter the effects of therapeutic interventions intended to increase subglottic pressure such as tracheostomy tube occlusion and decannulation.

Support

UPMC Competitive Medical Research Fund, Greater Pittsburgh Chapter, National Parkinson's Foundation.