CHD4 acts as a prognostic factor and drives radioresistance in HPV negative HNSCC.

Despite great efforts in improving existing therapies, the outcome of patients with advanced radioresistant HPV-negative head and neck squamous cell carcinoma (HNSCC) remains poor. The chromatin remodeler Chromodomain helicase DNA binding protein 4 (CHD4) is involved in different DNA-repair mechanisms, but the role and potential in HNSCC has not been explored yet. In the present study, we evaluated the prognostic significance of CHD4 expression using in silico analysis of the pan-cancer dataset. Furthermore, we established a monoclonal HNSCC CHD4 knockdown cell clone utilizing the CRISPR/Cas9 system. Effects of lower CHD4 expression on radiosensitivity after increasing doses of ionizing radiation were characterized using clonogenic assays and cell numbers. The in silico analysis revealed that high CHD4 expression is associated with significant poorer overall survival of HPV-negative HNSCC patients. Additionally, the knockdown of CHD4 significantly increased the radiosensitivity of HNSCC cells. Therefore, CHD4 might be involved in promoting radioresistance in hard-to-treat HPV-negative HNSCC entities. We conclude that CHD4 could serve as a prognostic factor in HPV-negative HNSCC tumors and is a potential target protein overcoming radioresistance in HNSCC. Our results and the newly established cell clone laid the foundation to further characterize the underlying mechanisms and ultimately use CHD4 in HNSCC therapies.

Comparison of a high-definition three-dimensional digital camera system with a conventional state-of-the-art operation microscope for microsurgical anastomoses.

Since its clinical implementation, microvascular surgery has depended on the continuous improvement of magnification tools. One of the more recent developments is a high-definition three-dimensional (3D) digital system (exoscope), which provides an alternative to the state-of-the-art operating microscopes. This study aimed to evaluate the advantages and disadvantages of this technology and compare it with its predecessor. The study included 14 surgeons with varying levels of experience, none of which had used a 3D optical system previously. Six of these surgeons performed five arterial and five venous anastomoses in the chicken thigh model with both the VITOM 3D exoscope-guided system and the Pentero operating microscope. These anastomoses were then evaluated for their quality and anastomosis time. The participants and the other eight surgeons, who had used the digital 3D camera system for microsurgical training exercises and vascular sutures, answered a questionnaire. The anastomosis time and number of complications were lower with the conventional microscope. Participants rated the image quality with the conventional microscope as higher, whereas the field of view and ergonomics were favorable in the digital 3D camera system. Exoscopes are optics suitable for performing simple microvascular procedures and are superior to classical microscopes ergonomically. Thus far, they are inferior to classical microscopes in terms of image quality and 3D imaging.

Protocol for generating monoclonal CRISPR-Cas9-mediated knockout cell lines using RNPs and lipofection in HNSCC cells.

CRISPR-Cas9 is a powerful technology for accurate and optimizable genome editing. Here, we present a protocol for generating monoclonal knockout (KO) cell lines using CRISPR-Cas9, ribonucleoprotein complexes (RNPs), and lipofection in adherent HNSCC cells from start to finish. We describe steps for choosing the suitable guide and primer design, preparation of guide-RNA (gRNA), lipofection of RNP complexes in HN cells, and single-cell cloning with limiting dilution. We then detail PCR and DNA purification and the selection and verification of monoclonal KO cell lines.

Comparing TIMP-1 and Hsp70 in Blood and Saliva as Potential Prognostic Markers in HNSCC.

(1) Background: Currently, there is no clinically used liquid biomarker in head and neck squamous cell carcinoma (HNSCC) patients. One reason could be the limited shedding of tumor material in early disease stages. Molecular diagnostics assessing both blood and especially saliva could potentially improve the accuracy of biomarkers. In this prospective study, two markers, tissue inhibitor of metalloprotease-1 (TIMP-1) and heat shock protein 70 (Hsp70), were analyzed in HNSCC patients. The purpose of the study was to evaluate differences between saliva and serum as sample material. Further, their prognostic and predictive value and usefulness for early detection was assessed. (2) Methods: A total of 73 HNSCC patients were prospectively monitored by collecting blood and saliva before, during, and after therapy, as well as in the follow-up period between 2018 and 2021. In total, 212 serum and 194 saliva samples were collected. A control group consisting of 40 subjects (15 patients with local infections in the head and neck area and 25 without infections) were examined as well. The collected samples were evaluated for the two proteins by using an enzyme-linked immunosorbent assay (ELISA). (3) RESULTS: The TIMP-1 concentration correlated significantly in blood and saliva, whereas the Hsp70 concentration did not. Saliva TIMP-1 was significantly higher in tumor patients compared to the control group (p = 0.013). High pretreatment TIMP-1 saliva levels were associated with significantly poorer disease-free survival (DFS) (p = 0.02). A high saliva TIMP-1/Hsp70 ratio was significantly associated with poorer DFS (HR: 1.4; 95% CI: 1.04-1.88; p = 0.026) and a high TIMP-1 serum concentration was significantly associated with poorer PFS (HR: 1.9; 95% CI: 1.2, 2.8; p = 0.003) and poorer overall survival (OS) (HR: 2.9; 95% CI: 1.4, 5.9; p = 0.003) in the Cox proportional hazards model. The saliva TIMP-1 to Hsp70 ratio was significantly higher at the time of recurrence (p = 0.015). Conclusion: TIMP-1 in serum is a promising prognostic marker for HNSCC. Saliva TIMP-1 and the saliva TIMP-1 to Hsp70 ratio provides additional information on the disease-free survival.

E-scooter-related craniomaxillofacial injuries compared with bicycle-related injuries - A retrospective study.

The aim of the study was to compare incidences and types of injury incurred between e-scooter crashes and bicycle crashes. This retrospective study included all victims of e-scooter crashes who were treated in the department of oral and maxillofacial surgery during a 12-months interval.A comparison was made with a cohort of patients who underwent bicycle crashes. Study parameters included type of fracture, soft tissue and dental affection, necessity of inpatient or outpatient treatment, the use of helmets, and the time of admission. In total, 400 patients were included. Of these 40 had suffered a crash on an e-scooter and 360 on a bicycle. Descriptive statistics showed a low helmet-wearing rate among cyclists (16.1% of recorded cases), with no helmet wearing recorded among e-scooter users. E-scooter-related crashes showed a higher rate of facial soft-tissue injuries (77.5%, p = 0.049) than among cyclists (61.7%), as well as a higher rate of dental injuries (27.5%, p = 0.017) compared with the bicycle cohort (13.3%). Facial fractures were also more common in the e-scooter cohort (45% vs 25.8%, p = 0.010). Admission was typically at the weekend - in the afternoon for the bicycle cohort and in the evening and at night for the e-scooter cohort. As a consequence of the fact that e-scooter riders seem to have an increased risk of facial injuries, it seems that an awareness campaign might be necessary to encourage helmet usage and to persuade intoxicated persons to use public transportation instead.

Temporal Tendinitis in Craniomandibular Dysfunction (CMD) - Does it Really Exist? A Temporomandibular MRI Investigation.

OBJECTIVES: The aim of the study was to analyze the role of temporal muscle and particularly tendon pathology in patients suffering from craniomandibular dysfunction (CMD) using magnetic resonance imaging. MATERIALS AND METHODS: Retrospective analysis of MRI examinations was carried out with regard to temporal muscle fibrosis and fatty degeneration and particularly temporal tendon rupture, thickening, and degenerative signal alterations. Descriptive statistics and the Mann-Whitney U-test were used for statistical evaluation. RESULTS: Structural lesions of temporal muscle parenchyma were the absolute exception. PD hyperintensity, pronounced contrast enhancement, or peritendinous fluid collections along the temporal tendon were found only to a small extent, and a (partial) rupture occurred in only one case. The tendon diameter showed only slight variability. The Mann-Whitney U-test provided no results indicating a causal connection between degenerative joint or disc disease and temporal tendon pathology. CONCLUSION: A large sample of 128 magnetic resonance imaging examinations provided no evidence of a major role of temporal tendinitis in clinical CMD syndrome. KEY POINTS: · Retrospective analysis of temporal tendon in CMD patients.. · Abnormal structural findings along the tendon seen only rarely.. · Obviously no crucial role of temporal tendon lesions in CMD syndrome.. CITATION FORMAT: · Stimmer H, Grill F, Waschulzik B et al. Temporal Tendinitis in Craniomandibular Dysfunction (CMD) - Does it Really Exist? A Temporomandibular MRI Investigation. Fortschr Röntgenstr 2022; 194: 1242 - 1249.

In-House, Open-Source 3D-Software-Based, CAD/CAM-Planned Mandibular Reconstructions in 20 Consecutive Free Fibula Flap Cases: An Explorative Cross-Sectional Study With Three-Dimensional Performance Analysis.

BACKGROUND: Mandibular reconstruction is conventionally performed freehand, CAD/CAM-assisted, or by using partially adjustable resection aids. CAD/CAM-assisted reconstructions are usually done in cooperation with osteosynthesis manufacturers, which entails additional costs and longer lead time. The purpose of this study is to analyze an in-house, open-source software-based solution for virtual planning. METHODS AND MATERIALS: All consecutive cases between January 2019 and April 2021 that underwent in-house, software-based (Blender) mandibular reconstruction with a free fibula flap (FFF) were included in this cross-sectional study. The pre- and postoperative Digital Imaging and Com munications in Medicine (DICOM) data were converted to standard tessellation language (STL) files. In addition to documenting general information (sex, age, indication for surgery, extent of resection, number of segments, duration of surgery, and ischemia time), conventional measurements and three-dimensional analysis methods (root mean square error [RMSE], mean surface distance [MSD], and Hausdorff distance [HD]) were used. RESULTS: Twenty consecutive cases were enrolled. Three-dimensional analysis of preoperative and virtually planned neomandibula models was associated with a median RMSE of 1.4 (0.4-7.2), MSD of 0.3 (-0.1-2.9), and HD of 0.7 (0.1-3.1). Three-dimensional comparison of preoperative and postoperative models showed a median RMSE of 2.2 (1.5-11.1), MSD of 0.5 (-0.6-6.1), and HD of 1.5 (1.1-6.5) and the differences were significantly different for RMSE (p < 0.001) and HD (p < 0.001). The difference was not significantly different for MSD (p = 0.554). Three-dimensional analysis of virtual and postoperative models had a median RMSE of 2.3 (1.3-10.7), MSD of -0.1 (-1.0-5.6), and HD of 1.7 (0.1-5.9). CONCLUSIONS: Open-source software-based in-house planning is a feasible, inexpensive, and fast method that enables accurate reconstructions. Additionally, it is excellent for teaching purposes.

The impact of intraoperative frozen section analysis on final resection margin status, recurrence, and patient outcome with oral squamous cell carcinoma.

BACKGROUND: The objective of this study was to evaluate the diagnostic value of intraoperative frozen section analysis (IFSA) of tumor bed margins in patients with oral squamous cell carcinoma (OSCC). METHODS: This retrospective study includes 194 primary OSCC cases. The impact of intraoperative information by IFSA on final margin status, local recurrence, and disease-specific survival were analyzed. RESULTS: IFSA revealed a 50% sensitivity and a 100% specificity, with a positive and negative predictive value of 100% and 89.1%, respectively. In 19 cases, margins were rated positive by IFSA and remained positive in eight cases (42.1%), despite immediate re-resection. This constellation led to higher recurrence and lower survival rates than in cases with consecutive R0 status (each p = 0.046). Positive margins in IFSA were associated with closer final margins (p = 0.022) and early recurrences (p = 0.050). CONCLUSIONS: Achieving instant R0 status has a crucial impact on disease recurrence and patient survival. IFSA falls short to ensure secure definite surgical margins. Thus, improved intraoperative diagnostic information on the location and extent of OSCC could support patient treatment. CLINICAL RELEVANCE: Considering that patient survival has not improved despite progress in surgical and adjuvant therapy, the process and outcome of IFSA was scrutinized as one part of the treatment concept.

The organometallic ferrocene exhibits amplified anti-tumor activity by targeted delivery via highly selective ligands to αvß3, αvß6, or α5ß1 integrins.

High levels of reactive oxygen species (ROS) in tumors have been shown to exert anti-tumor activity, leading to the concept of ROS induction as therapeutic strategy. The organometallic compound ferrocene (Fc) generates ROS through a reversible one-electron oxidation. Incorporation of Fc into a tumor-targeting, bioactive molecule can enhance its therapeutic activity and enable tumor specific delivery. Therefore, we conjugated Fc to five synthetic, Arg-Gly-Asp (RGD)-based integrin binding ligands to enable targeting of the cell adhesion and signaling receptor integrin subtypes αvß3, α5ß1, or αvß6, which are overexpressed in various, distinct tumors. We designed and synthesized a library of integrin-ligand-ferrocene (ILF) derivatives and showed that ILF conjugates maintained the high integrin affinity and selectivity of their parent ligands. A thorough biological characterization allowed us to identify the two most promising ligands, an αvß3 (L2b) and an αvß6 (L3b) targeting ILF, which displayed selective integrin-dependent cell uptake and pronounced ferrocene-mediated anti-tumor effects in vitro, along with increased ROS production and DNA damage. Hence, ILFs are promising candidates for the selective, tumor-targeted delivery of ferrocene to maximize its anti-cancer efficacy and minimize systemic toxicity, thereby improving the therapeutic window of ferrocene compared to currently used non-selective anti-cancer drugs.

Defining secure surgical bone margins in head and neck squamous cell carcinomas: The diagnostic impact of intraoperative cytological assessment of bone resection margins compared with preoperative imaging.

BACKGROUND: Imaging provides crucial staging information for treatment planning of head and neck squamous cell carcinomas (HNSCCs). Despite technical progress in imaging techniques, defining the extent of bone involvement preoperatively remains challenging and requires intraoperative information to control for adequate resection. The intraoperative cytological assessment of the bone resection margins (ICAB) provides information whether bone is infiltrated by carcinoma. The aim of this study was to assess the diagnostic value of preoperative imaging compared with ICAB in order to achieve carcinoma-free bone margins. MATERIALS AND METHODS: 108 HNSCC patients underwent preoperative computed tomography (CT), magnetic resonance imaging (MRI) and orthopantomogram (OPG) for staging and surgical planning. Curative resection was planned based on imaging. Intraoperatively, the resection margins were controlled by ICAB. The diagnostic value of preoperative imaging and ICAB was assessed with reference to the histological findings. RESULTS: CT showed a sensitivity of 89.7%, specificity of 63.0%, positive predictive value (PPV) of 85.9%, and negative predictive value (NPV) of 70.8%. MRI revealed a sensitivity of 45.5%, specificity of 66.7%, PPV of 71.4% and NPV of 40.0%. OPG-imaging had a sensitivity of 64.7%, specificity of 76.2%, PPV of 81.5%, NPV 57.1%. In comparison, ICAB provided a sensitivity of 78.6%, specificity of 95.7%, PPV 73.3%, and NPV 96.7%. The accuracy was 82.1%, 52.9%, 69.0%, and 93.5% for CT, MRI, OPG, and ICAB, respectively. CONCLUSION: Preoperative imaging lacks accuracy in defining adequate bone resection margins, compared with ICAB. ICAB supports preoperative imaging and intraoperative frozen sections to improve bone margin control.

Multi-Parametric Standardization of Fluorescence Imaging Systems Based on a Composite Phantom.

OBJECTIVE: Fluorescence molecular imaging (FMI) has emerged as a promising tool for surgical guidance in oncology, with one of the few remaining challenges being the ability to offer quality control and data referencing. This paper investigates the use of a novel composite phantom to correct and benchmark FMI systems. METHODS: This paper extends on previous work by describing a phantom design that can provide a more complete assessment of FMI systems through quantification of dynamic range and determination of spatial illumination patterns for both reflectance and fluorescence imaging. Various performance metrics are combined into a robust and descriptive "system benchmarking score," enabling not only the comprehensive comparison of different systems, but also for the first time, correction of the acquired data. RESULTS: We show that systems developed for targeted fluorescence imaging can achieve benchmarking scores of up to 70%, while clinically available systems optimized for indocyanine green are limited to 50%, mostly due to greater leakage of ambient and excitation illumination and lower resolution. The image uniformity can also be approximated and employed for image flat-fielding, an important milestone toward data referencing. In addition, we demonstrate composite phantom use in assessing the performance of a surgical microscope and of a raster-scan imaging system. CONCLUSION: Our results suggest that the new phantom has the potential to support high-fidelity FMI through benchmarking and image correction. SIGNIFICANCE: Standardization of the FMI is a necessary process for establishing good imaging practices in clinical environments and for enabling high-fidelity imaging across patients and multi-center imaging studies.

Selective Targeting of Integrin αvß8 by a Highly Active Cyclic Peptide.

Integrins play important roles in physiological and pathophysiological processes. Among the RGD-recognizing integrin subtypes, the αvß8 receptor is emerging as an attractive target because of its involvement in various illnesses, such as autoimmune diseases, viral infections, and cancer. However, its functions have, so far, not been investigated in living subjects mainly because of the lack of a selective αvß8 ligand. Here, we report the design and potential medical applications of a cyclic octapeptide as the first highly selective small-molecule ligand for αvß8. Remarkably, this compound displays low nanomolar αvß8 binding affinity and a strong discriminating power of at least 2 orders of magnitude versus other RGD-recognizing integrins. Peptide functionalization with fluorescent or radioactive labels enables the selective imaging of αvß8-positive cells and tissues. This new probe will pave the way for detailed characterization of the distinct (patho)physiological role of this relatively unexplored integrin, providing a basis to fully exploit the potential of αvß8 as a target for molecular diagnostics and personalized therapy regimens.

Exploring the Role of RGD-Recognizing Integrins in Cancer.

Integrins are key regulators of communication between cells and with their microenvironment. Eight members of the integrin superfamily recognize the tripeptide motif Arg-Gly-Asp (RGD) within extracelluar matrix (ECM) proteins. These integrins constitute an important subfamily and play a major role in cancer progression and metastasis via their tumor biological functions. Such transmembrane adhesion and signaling receptors are thus recognized as promising and well accessible targets for novel diagnostic and therapeutic applications for directly attacking cancer cells and their fatal microenvironment. Recently, specific small peptidic and peptidomimetic ligands as well as antibodies binding to distinct integrin subtypes have been developed and synthesized as new drug candidates for cancer treatment. Understanding the distinct functions and interplay of integrin subtypes is a prerequisite for selective intervention in integrin-mediated diseases. Integrin subtype-specific ligands labelled with radioisotopes or fluorescent molecules allows the characterization of the integrin patterns in vivo and later the medical intervention via subtype specific drugs. The coating of nanoparticles, larger proteins, or encapsulating agents by integrin ligands are being explored to guide cytotoxic reagents directly to the cancer cell surface. These ligands are currently under investigation in clinical studies for their efficacy in interference with tumor cell adhesion, migration/invasion, proliferation, signaling, and survival, opening new treatment approaches in personalized medicine.

Effect of an evidence-based guideline on the treatment of maxillofacial cancer: A prospective analysis.

BACKGROUND: In 2012, a guideline for the diagnosis and treatment of oral cavity cancer based on the best available evidence was implemented at certified German cancer centres for head and neck carcinomas. The present analysis was performed to determine whether the implementation of the guideline via certification improved the level of care, leading to a benefit for the patients. METHODS: A prospective observational study was performed based on the annual operating figures at 31 certified head and neck cancer centres. From 76 statements and recommendations, 9 indicators were derived defining important steps during treatment. The annual shift of the figures was documented for each indicator and was used to measure the impact of the guideline. This was achieved by determining the number of patients having received the recommended treatment related to the total number in each centre over a period of 3 years. RESULTS: In 2014, 1570 primary cases with an oral cavity carcinoma were treated at our centres, 31.2% representing stage IVA. Except for two, all indicators showed increasing numbers of achievement from 2012 to 2014, reaching median values between 91% and 100% in 2014. In particular, median values for imaging and interdisciplinary treatment to evaluate the presence of second primaries and metastases increased by 20% and 30%, respectively. Median values decreased by 14% for recommended adjuvant radiation, because of non-acceptance by the patients. Moreover, elective neck dissection was performed less frequently in cN0 categories. CONCLUSIONS: Implementation of the national cancer guideline by means of certification evidently had a positive impact on patients suffering from oral squamous cell carcinoma and led to the improved achievement of most evidence-based treatment recommendations over time. Further research involving high-level clinical studies is needed to cover all aspects of this specific tumour entity.

Clinical Impact of Intraoperative Cytological Assessment of Bone Resection Margins in Patients with Head and Neck Carcinoma.

BACKGROUND: Surgical treatment of head and neck squamous cell carcinoma (HNSCC) patients often results in complex defects, affecting functional structures. Frozen sections are valuable to guide resections and control for adequate margins; however, intraoperative assessment of bone remains challenging. OBJECTIVE: The objective of this study was to evaluate the clinical impact of an intraoperative cytological assessment of bone margins (ICAB) on resection status and patient outcome. METHODS: ICAB analysis (n = 267) was implemented in 102 patients during resection of HNSCC for a guided resection of affected bone. The cytological findings were compared with the final histological results of the corresponding bone margins, and the surgical consequences, R1 rates, and patient outcome of the ICAB intervention group were compared with an equal control group of 100 patients. RESULTS: ICAB revealed a sensitivity of 94.4 % [95 % confidence interval (CI) 81.3-99.3], specificity of 97.4 % (95 % CI 94.4-99.0), positive predictive value of 85.0 % (95 % CI 70.2-94.3), and negative predictive value of 99.1 % (95 % CI 96.9-99.9). Osseous R1 resections were reduced from 8 to 2.9 % (∆R1 = 5.1 %; p = 0.113), rendering a relative risk reduction (RRR) of 63.2 % with a number needed to treat (NNT) of 19.57. ICAB influenced final resection status, with a reduction of R1 resections from 17 to 7.8 % (∆R1 = 9.2 %; p = 0.026), with an RRR of 59.65 % and an NNT of 9.66. The ICAB intervention group revealed a higher disease-free survival [p(log-rank) = 0.045] and overall survival [p(log-rank) = 0.014] according to multivariable analysis. CONCLUSION: ICAB, applied as a routine diagnostic tool to supplement frozen sections, can help to reduce R1 resections in order to improve patient outcome.

Stable Peptides Instead of Stapled Peptides: Highly Potent αvß6-Selective Integrin Ligands.

The αvß6 integrin binds the RGD-containing peptide of the foot and mouth disease virus with high selectivity. In this study, the long binding helix of this ligand was downsized to an enzymatically stable cyclic peptide endowed with sub-nanomolar binding affinity toward the αvß6 receptor and remarkable selectivity against other integrins. Computational studies were performed to disclose the molecular bases underlying the high binding affinity and receptor subtype selectivity of this peptide. Finally, the utility of the ligand for use in biomedical studies was also demonstrated here.

Oral squamous cell carcinoma: the impact of stage-dependent therapy regimes on postoperative disease recurrence.

OBJECTIVE: To investigate disease recurrence among patients with oral squamous cell carcinoma (OSCC) and positive lymph node lesions receiving different therapy for different stages of the disease. STUDY DESIGN: One hundred sixty-one patients with biopsy-proven pN1 and pN2+ carcinomas were reviewed retrospectively. We investigated postoperative disease recurrence when using surgery alone (SA), adjuvant radiotherapy (RT), or adjuvant radiochemotherapy (RCT) and compared our results with the 2012 German guidelines. RESULTS: Among the study population, 60 (37%) experienced postoperative disease recurrence, 38% with SA, 42% with adjuvant RT, and 39% with adjuvant RCT (P = .420). In the case of pN1, the recurrence rate was 14% with SA, 32% with RT, and 33% with RCT (P = .156); and in pN2+, 60%, 49%, and 29%, respectively (P = .012). CONCLUSIONS: Surgery plus adjuvant therapy has benefits with respect to postoperative disease recurrence in pN2+ OSCC. In pN1, surgery alone might be as effective as adjuvant RT or RCT.

Immunolocalization of antimicrobial and cytoskeletal components in the serous glands of human sinonasal mucosa.

Secretory cells in the seromucous glands of paranasal sinuses secrete antibacterial proteins for innate immune mucosal integrity. We studied the localization of antimicrobial and cytoskeletal components of the human seromucous glands and respiratory epithelium of the maxillary sinus and the ethmoidal cells by immunohistochemical methods. The presence of a variety of defense proteins such as lysozyme, lactoferrin, cathelicidin, and defensin-1, -2, -3 point to a crucial role in the immune defense for the respiratory tract. Cytoskeletal proteins such as actin, myosin 2, cytokeratin 7 and 19, α- and ß-tubulin, investigated for the first time in glands of paranasal sinuses, showed a stronger expression at the apical and lateral cell membrane. The localization of the cytoskeletal proteins might point to their participation in exocrine secretory processes and stabilizing effects.

Evaluation of intraoperative cytological assessment of bone resection margins in patients with oral squamous cell carcinoma.

BACKGROUND: Frozen sections are routinely applied to control for adequate resection margins. In cases in which carcinoma infiltrates bone, the intraoperative microscopic assessment of bone margins remains challenging due to technical difficulties to section native bone. The objective of the current study was to evaluate an intraoperative cytological approach to control bone resection margins in patients with bone-infiltrating oral squamous cell carcinomas. METHODS: A total of 174 cytological preparations obtained from bone margins of bone-infiltrating oral squamous cell carcinomas (28 patients) were assessed intraoperatively and compared with the corresponding histological findings. In a validation cohort (45 patients) the intraoperative cytological assessment of bone resection margins (ICAB) (104 margins) was evaluated as a diagnostic tool for routine clinical application. RESULTS: In the first patient cohort, the ICAB revealed 95.3% sensitivity and 96% specificity. The results provided an accuracy of 95.7% with a significant correlation noted between cytological and histological results (κ, 0.91; P < .001), and a positive predictive value and negative predictive value of 93.8% and 96.9%, respectively. In the validation cohort, ICAB revealed 80% sensitivity and 98.9% specificity with 98% accuracy. There was a significant correlation found between cytological and histological results (κ, 0.91; P < .001), providing a positive predictive value and negative predictive value of 80% and 98%, respectively. ICAB could predict final resection status at bone margins with 80% sensitivity and 97.5% specificity. A significant correlation was found between the cytological and histological resection status at bone margins (κ, 0,75; P < .001). CONCLUSIONS: ICAB could supplement intraoperative frozen sections of soft tissue margins as a standard procedure to control for adequate resection at bone margins.

Human ß-defensins and psoriasin/S100A7 expression in salivary glands: anti-oncogenic molecules for potential therapeutic approaches.

BACKGROUND: Host defence peptides (HDPs), including human ß-defensins (hBDs) and psoriasin/S100A7, exert antimicrobial and immunoregulatory functions of the innate defense system. In addition to these functions, the search for cancer biomarkers has identified HDPs as playing a potential role in both tumor suppression and oncogenesis. Although HDPs are highly expressed in salivary glands, their role as molecules for potential diagnostic and therapeutic approaches has not yet been analyzed. OBJECTIVE: The aim of the present study was to investigate whether expression levels of putative pro- or anti-oncogenic hBDs, including hBD-1, -2, -3, and psoriasin/S100A7, are altered in salivary gland tumor tissue as potential targets for molecular-based therapeutic approaches. METHODS: We analyzed the expression levels of hBD-1, -2, -3, and psoriasin/S100A7 by quantitative real-time polymerase chain reaction (qrt-PCR) and immunohistochemistry in a case control study by comparing salivary gland tumor samples relative to healthy control specimens from 58 patients. Expression level analysis of hBD-1, -2, -3, and psoriasin/S100A7 by qrt-PCR was normalized to the endogenous 18S rRNA expression levels. RESULTS: The results demonstrate the significant downregulation of hBD-1 (p < 0.001), hBD-2 (p = 0.003), hBD-3 (p = 0.002), and psoriasin/S100A7 (p = 0.003) mRNA in human salivary gland tumors compared with healthy control specimens. Protein expression levels of hBD-1, -2, -3, and psoriasin/S100A7 in salivary gland tumor tissue were strongly reduced compared with healthy control specimens. CONCLUSION: The data indicates a putative role of the innate defense system in salivary gland tumor formation. The identification of immunoregulatory molecules as diagnostic biomarkers or therapeutic targets could provide new approaches for molecular-based diagnostic and therapeutic support to treat salivary gland tumors as well as other malignancies. We suggest that HDPs should be taken into consideration for use in molecular-based therapeutic approaches.