bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

ProblemThe 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.MethodsMSC 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.ResultsThe 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.ConclusionIntegrin‐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.SignificanceTo 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

ProblemTransforming 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.MethodsThe 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.ResultsImmunohistochemical 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.ConclusionTGF‐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.SignificanceThe 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

ProblemA 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.Methods3D 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.ResultsUsing 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.ConclusionMRI 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.SignificanceThis 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.SupportNIH‐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

ProblemA 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.MethodsIn 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.ResultsConsidering 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%.ConclusionA group of clinical variables in multiparametric combinations with molecular alterations discriminated early and late stage HNSCC. CART® improved the model's performance.SignificanceValidation 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.SupportNIH 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

ProblemAngiogenesis 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.MethodsEstablished 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.ResultsIn 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.ConclusionThe 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.SignificanceOur 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

ProblemThis 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.MethodsFive 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.ResultsAfter 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.ConclusionThe 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.SignificanceA possible alternative for primary screening of NPC.SupportNSC 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

ProblemTo determine the correlation between IL10, a Th2‐type inhibitory cytokine, clinical outcome and survival in HNSCC patients.MethodsIL10 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.ResultsBoth 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.ConclusionIL10 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.SignificanceIL10 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

ProblemQuantitate the intratumoral differentiation of HNSCC induced immunoinhibitory CD34+ in response to enteral treatment of 1,25 dihydroxyvitamin D3 (1,25D3).Methods8 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.ResultsAfter 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.ConclusionCD34+ 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.SignificanceDespite 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.SupportClinical 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

ProblemWe 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.MethodsParaffin‐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.ResultsHierarchical 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).ConclusionWe 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.SignificanceThese findings may have significant implications in patient selection for HPV‐specific therapies.SupportAAO/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

ProblemTo 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.MethodsParaffin 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.Results2 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.ConclusionLymphoscintigraphy 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.SignificanceBy 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.SupportResident 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

ProblemSquamous 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.MethodsTumor 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).ResultsFrom 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.ConclusionUsing two validated, complementary proteomics methods we identified no significant differences between HNSCCs from different head and neck subsites.SignificanceIt 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).SupportNIHR33‐CA‐5941(WSD); AAO/HNSF Resident Research Grant 26357 (PMW).