Comparison of Whiskbroom and Pushbroom darkfield elastic light scattering spectroscopic imaging for head and neck cancer identification in a mouse model.

The early detection of head and neck cancer is a prolonged challenging task. It requires a precise and accurate identification of tissue alterations as well as a distinct discrimination of cancerous from healthy tissue areas. A novel approach for this purpose uses microspectroscopic techniques with special focus on hyperspectral imaging (HSI) methods. Our proof-of-principle study presents the implementation and application of darkfield elastic light scattering spectroscopy (DF ELSS) as a non-destructive, high-resolution, and fast imaging modality to distinguish lingual healthy from altered tissue regions in a mouse model. The main aspect of our study deals with the comparison of two varying HSI detection principles, which are a point-by-point and line scanning imaging, and whether one might be more appropriate in differentiating several tissue types. Statistical models are formed by deploying a principal component analysis (PCA) with the Bayesian discriminant analysis (DA) on the elastic light scattering (ELS) spectra. Overall accuracy, sensitivity, and precision values of 98% are achieved for both models whereas the overall specificity results in 99%. An additional classification of model-unknown ELS spectra is performed. The predictions are verified with histopathological evaluations of identical HE-stained tissue areas to prove the model's capability of tissue distinction. In the context of our proof-of-principle study, we assess the Pushbroom PCA-DA model to be more suitable for tissue type differentiations and thus tissue classification. In addition to the HE-examination in head and neck cancer diagnosis, the usage of HSI-based statistical models might be conceivable in a daily clinical routine.

Evaluation of digital image analysis as a supportive tool for the stratification of head and neck vascular anomalies.

BACKGROUND: The histological differentiation of individual types of vascular anomalies (VA), such as lymphatic malformations (LM), hemangioma (Hem), paraganglioma (PG), venous malformations (VeM), arteriovenous malformations (AVM), pyogenic granulomas (GP), and (not otherwise classified) vascular malformations (VM n.o.c.) is frequently difficult due to the heterogeneity of these anomalies. The aim of the study was to evaluate digital image analysis as a method for VA stratification METHODS: A total of 40 VA tissues were examined immunohistologically using a selection of five vascular endothelial-associated markers (CD31, CD34, CLDN5, PDPN, VIM). The staining results were documented microscopically followed by digital image analyses based quantification of the candidate-marker-proteins using the open source program ImageJ/Fiji. RESULTS: Differences in the expression patterns of the candidate proteins could be detected particularly when deploying the quotient of the quantified immunohistochemical signal values. Deploying signal marker quotients, LM could be fully distinguished from all other tested tissue types. GP achieved stratification from LM, Hem, VM, PG and AVM tissues, whereas Hem, PG, VM and AVM exhibited significantly different signal marker quotients compared with LM and GP tissues. CONCLUSION: Although stratification of different VA from each other was only achieved in part with the markers used, the results of this study strongly support the usefulness of digital image analysis for the stratification of VA. Against the background of upcoming new diagnostic techniques involving artificial intelligence and deep (machine) learning, our data serve as a paradigm of how digital evaluation methods can be deployed to support diagnostic decision making in the field of VAs.

Two giant retroperitoneal schwannomas mimicking adrenal malignancy - a case report.

Schwannomas are benign tumors in 95% of cases and very rarely occur in the retroperitoneum. We report the cases of a 35-year-old man with abdominal discomfort and a 50-year-old asymptomatic woman with large retroperitoneal masses. Both underwent multivisceral surgery to exclude an adrenal carcinoma, and the pathologic diagnosis showed schwannomas in both cases. Despite morphological imaging, it was not possible to get a clear diagnosis preoperatively.

CD73 expression in lymph node metastases in patients with head and neck cancer.

INTRODUCTION: CD73 is an enzyme crucial in the metabolism of immunosuppressive adenosine. In cancer, it has various functions including tumor growth and metastases. Squamous cell carcinoma of the head and neck has an overall poor prognosis, also due to early spread of metastatic cells. MATERIALS AND METHODS: Tumor and lymph node specimens of 65 patients with HNSCC were subjected to immunohistochemical and H-score analysis of CD73 expression. Demographics, diagnoses, histopathology and subsequent outcome were analyzed. RESULTS: The primary cancer was squamous cell carcinoma in all patients (male/female 55:10) with the following locations: oral cavity n:16, oropharynx n:28, hypopharynx n:11 and larynx n:10. H-score for CD73 expression in the primary lesion and metastatic lymph nodes was significantly higher in advanced compared to early stages with no significant differences among tumor locations. High CD73 expression was associated with reduced overall survival rates at a mean follow-up of 83.4 months (6-204 months). CONCLUSIONS: CD73 expression in HNSCC correlated positively with tumor stage and was associated with poor prognosis. Therefore, CD73 expression in primary lesions and regional metastases appears to predict HNSCC patients at high risk of all tumor sites. Therapeutic approaches targeting CD73 might seem promising for this patient population.

ADAM8 expression in breast cancer derived brain metastases: Functional implications on MMP-9 expression and transendothelial migration in breast cancer cells.

Metastatic breast cancer affects long-term survival and is a major cause of cancer death for women worldwide. The Metalloprotease-Disintegrin ADAM8 promotes breast cancer development and brain metastasis in a mouse breast cancer model. Here, abundant ADAM8 expression was detected in primary human breast tumors and associated brain metastases. To investigate the function of ADAM8 in metastasis, MB-231 breast cancer cells with ADAM8 knockdown (MB-231_shA8) and scramble control cells (MB-231_shCtrl) were analyzed for their capability to develop metastases. In vitro, formation of metastatic complexes in hanging drops is dependent on ADAM8 and blocked by ADAM8 inhibition. MB-231_shA8 in contrast to MB-231_shCtrl cells were impaired in transmigration through an endothelial and a reconstituted blood-brain barrier. Out of 23 MMP and 22 ADAM genes, only the MMP-9 gene was affected by ADAM8 knockdown in MB-231_shA8 cells. Following re-expression of wild-type ADAM8 in contrast to ADAM8 lacking the cytoplasmic domain in MB-231_shA8 cells caused increased levels of activated pERK1/2 and pCREB (S133) that were associated with elevated MMP-9 transcription. Application of ADAM8 and MMP-9 antibodies reduced transmigration of MB-231 cells suggesting that ADAM8 affects transmigration of breast cancer cells by MMP-9 regulation. ADAM8-dependent transmigration was confirmed in Hs578t cells overexpressing ADAM8. Moreover, transmigration of MB-231 and Hs578t cells was significantly reduced for cells treated with an antibody directed against P-selectin glycoprotein ligand (PSGL-1), a substrate of ADAM8. From these data we conclude that ADAM8 promotes early metastatic processes such as transendothelial migration by upregulation of MMP-9 and shedding of PSGL-1 from breast cancer cells.

Ectonucleotidase CD39 expression in regional metastases in head and neck cancer.

INTRODUCTION: CD39 is the rate-limiting enzyme in the generation of immunosuppressive adenosine and its expression and activity are significant in tumor progression. Squamous cell carcinoma of the head and neck (HNSCC) shows an overall poor prognosis due to high local recurrence rates and early metastatic spread. MATERIAL AND METHODS: Primary tumor specimens and lymph node specimens harvested during neck dissection of 65 patients with a diagnosis of HNSCC were subjected to immunohistochemical and H-score analysis of CD39 expression. Demographics, histopathology and subsequent outcome were analyzed. RESULTS: The primary cancer was squamous cell carcinoma in all patients (male/female 55:10). H-score for CD39 expression in the primary lesion and metastatic lymph nodes was significantly higher in advanced compared to early stages with no significant differences among different tumor locations. High intratumoral and intrametastatic CD39 expression was associated with an inferior patients' overall survival at a mean follow-up of 83.4 months (6-204 months). CONCLUSION: CD39 expression in HNSCC correlated positively with tumor stage and appears to predict poor prognosis. Therefore, CD39 expression in primary lesions and metastatic lymph nodes seems to identify patients at high risk in HNSCC of all tumor sites. Immunotherapeutic approaches targeting CD39 might be promising for this patient population.

Foxp3 expression in lymph node metastases in patients with head and neck cancer.

INTRODUCTION: The prevalence and activity of regulatory T cells in patients with cancer correlates with poor prognosis. These cells are characterized by their expression of Forkhead box protein-3 (Foxp3). Squamous cell carcinoma is the most prevalent type of cancer in the head and neck region with overall poor survival rates, also due to early spread of metastatic cells. MATERIAL AND METHODS: Primary tumor specimens as well as lymph node specimens harvested during neck dissection of 65 patients with a diagnosis of HNSCC were subjected to immunohistochemical and H-score analysis of Foxp3 expression. Demographics, diagnoses, histopathology and subsequent outcome were analyzed. RESULTS: The primary cancer was squamous cell carcinoma in all patients (male/female 55:10) with the following tumor locations: oral cavity n = 16, oropharynx n = 28, hypopharynx n = 11 and larynx n = 10 (Stage III n = 18; Stage IVA n = 45; Stage IVB n = 2). The H-score for Foxp3 expression in the primary lesion as well as metastatic lymph nodes was significantly higher in advanced stages compared to early stages with differences among tumor locations, which were not significant. High Foxp3 expression was associated with inferior overall survival rates at a mean follow-up of 83.4 months (6-204 months) Conclusions: Foxp3 expression in HNSCC varied from the anatomical site and correlated positively with tumor stage and was associated with poor prognosis. Therefore, Foxp3 expressions in primary lesions as well as lymphogenic metastases appear to predict high-risk HSNCC patients. Novel therapeutic approaches targeting Foxp3+ cells might seem promising for this patient population.

Freehand SPECT for sentinel lymph node detection in patients with head and neck cancer: first experiences.

CONCLUSION: Feasibility of intraoperative 3D imaging with freehand (fh) SPECT for sentinel lymph node (SLN) biopsy in head and neck cancer (HNC) could be demonstrated. Controlled clinical studies are needed to evaluate its accuracy and impact on patient morbidity. OBJECTIVES: The clinical N0 neck in HNC needs improvement in management to sufficiently detect occult neck disease but to spare patients from potential morbidity by elective neck dissection. The SLN concept has potential to accurately stage the neck with low morbidity. METHODS: fhSPECT is a 3D tomographic imaging modality with a gamma probe system combined with an infrared optical tracking system. Five patients with HNC and clinical N0 neck were recruited. Scanning for SLN using fhSPECT was performed before excision and selective neck dissection and specimens were analyzed histopathologically. RESULTS: Preoperatively, a total of nine SLNs were located in five patients with fhSPECT. SLNs in three patients were positive for metastatic disease; in two patients the SLNs were tumor-free. No residual radioactivity was found in the neck in any of the patients after extirpation of SLNs. fhSPECT acquisitions took 2.6 ± 0.4 min. No metastatic lymph nodes were detected in any other node harvested during subsequent selective neck dissection in any patients.

The chorioallantoic membrane assay is a promising ex vivo model system for the study of vascular anomalies.

UNLABELLED: The present feasibility study evaluated the chorioallantoic membrane (CAM) assay established in cancer and angiogenesis research as a tool for the study of vascular anomalies (VAs) in the head and neck area, since the lack of appropriate model systems poses a major obstacle in VA research. MATERIALS AND METHODS: VA tissues from three patients, two with an arteriovenous and one with a lymphatic malformation, were analyzed and evaluated in the CAM assay. RESULTS: The arteriovenous malformations induced a potent angiogenic reaction, resulting in new vessel growth and reperfusion by chicken embryo blood, which was comparable in extent with the positive vascular endothelial growth factor control. An angiogenic reaction, although less pronounced, was also observed in the single-tested lymphatic malformation. CONCLUSION: Our observations indicate the CAM assay to be a suitable model system for the study of VAs, as well as to show how treatment with pro- and antiangiogenic drugs affects VA growth patterns. The CAM assay has the potential to become a valuable tool for VA studies.

Human papilloma virus (HPV) is not implicated in the etiology of Warthin's tumor of the parotid gland.

CONCLUSION: The lack of human papilloma virus (HPV) sequences as well as potential HPV-activated cells such as cells that would be p16- and Ki-67 positive does not support a role of HPV in the pathogenesis of this lesion. OBJECTIVE: The exact etiopathogenesis of Warthin's tumor of the parotid gland is still unclear. The aim of the present study was to evaluate if HPV could play a role in the development of this parotid lesion. METHODS: Tissue samples from 40 Warthin's tumors of the parotid gland were investigated by PCR followed by in situ hybridization. The immunohistochemical expression of p16 and the dual immunostaining of p16 and Ki-67 were evaluated in all samples. RESULTS: Strong and diffuse p16 immunoreactivity was found in 7 of the 40 cases (17.5%). In situ hybridization showed a diffuse episomal signal in those samples. However, PCR could not reliably detect the presence of HPV genes. Furthermore, p16-expressing epithelial cells were mostly negative for the proliferation marker Ki-67.

Port-wine stains are more than skin-deep! Expanding the spectrum of extracutaneous manifestations of nevi flammei of the head and neck.

UNLABELLED: It is well known that port-wine stains of the upper part of the face may herald abnormalities of the brain or eye in the form of Sturge-Weber syndrome. This study focuses on other extracutaneous anomalies in patients with nevi flammei of the head and neck, giving rise to functional complications. PATIENTS AND METHODS: A retrospective study was performed on patients with port-wine stains involving the head and neck area. Records were reviewed for demographic parameters, extent of the lesion, clinical complications, diagnostic measures, previous treatments, ultimate therapeutic approach, and outcome. RESULTS: Nine patients, mean age 50.4 years, with port-wine stains and clinical symptoms due to extracutaneous involvement, were admitted and treated from 2006 to 2009. Major clinical features included macrocheilia in three cases, gingival bleeding in two, dysphonia with globus sensation, painful parotideal swelling with recurrent otitis, painful lingual swelling, recurrent epistaxis, and nasal obstruction in one case each. Cases with lower lip hypertrophy were treated by conventional surgical approaches. Recurrent epistaxis and nasal obstruction due to affected inferior turbinate were treated by Nd:YAG laser therapy, and globus sensation and dysphonia by speech therapy. Patients with gingival affection and recurrent otitis were treated by local ear care. CONCLUSION: Port-wine stains in the head and neck may develop extracutaneous manifestations causing severe problems. A multimodal and interdisciplinary approach is mandatory for an appropriate treatment.

Lack of significant estrogen and progesterone receptor expression in nasal telangiectasias in hereditary hemorrhagic telangiectasia: an immunohistochemical analysis.

CONCLUSION: This immunohistochemical study of estrogen and progesterone receptors could not confirm a significant expression in nasal telangiectasias. Thus, a specific effect of these hormones or anti-hormone therapy on malformed nasal vessels has to be questioned and only offered under strict clinical control. OBJECTIVE: The efforts to control recurrent epistaxis in hereditary hemorrhagic telangiectasia (HHT) using alternative methods are very intense. Hormone or anti-hormone therapy has frequently been postulated and the reported results are controversial. Therefore it was important to find an explanation regarding a possible impact of hormonal therapies by immunohistochemical evaluation of progesterone and estrogen receptor expression on nasal telangiectasias of affected patients. METHODS: Tissue samples of nasal mucosa with evidence of telangiectasias from 14 patients with HHT were analyzed for the expression of progesterone and estrogen receptors on the nuclei of endothelial cells of the malformed vessels using immunohistochemistry. RESULTS: Progesterone receptors were not detected in any of the cases and only two cases showed a weak expression of estrogen receptors with an immunoreactive score of 2/12.

Adult rhabdomyoma in the parapharyngeal space: report of 2 cases and review of the literature.

Rhabdomyomas are rare benign mesenchymal tumors with skeletal muscle differentiation. They are less commonly encountered than are their malignant counterparts, rhabdomyosarcomas. Rhabdomyomas fall into 2 general categories: cardiac and extracardiac types. Extracardiac rhabdomyomas are among the rarest tumors in humans and can be subclassified as fetal, juvenile, and adult types depending on the individual tumor's degree of differentiation by light microscopy. Adult extracardiac rhabdomyoma has a strong predilection for occurrence in the head and neck, mainly in the area of larynx and pharynx. In this article, 2 cases of parapharyngeal rhabdomyoma are reported together with a review of the world literature.

Intralesional endoscopy of advanced lymphatic malformations of the head and neck: a new diagnostic approach and a potential therapeutic tool.

OBJECTIVES: To evaluate the morphologic properties of advanced lymphatic malformations of the head and neck and to investigate possible therapeutic interventions by intralesional endoscopy. DESIGN: This case study analyzes the outcome of intralesional endoscopy of lymphatic malformations of the head and neck and discusses the results of this approach. SETTING: Academic medical center. PATIENTS: Endoscopy was performed on 4 patients with cervicofacial lymphatic malformations. INTERVENTIONS: Endoscopy of lymphatic malformations. RESULTS: The endoscopic inspection and assessment of the lesions revealed detailed information about morphologic aspects like the structure of intralesional septa, internal vascularization, and intercystic correspondence and channel networks. Anatomic landmarks could be followed on their intralesional courses. CONCLUSIONS: Intralesional endoscopy of lymphatic malformations provides insight into the morphologic properties of the lesion and is a useful supplementary tool during conventional surgical intervention and for sclerosing therapy. The technique allows a detailed assessment of these lesions and opens a broad spectrum of therapeutic options.

Is it necessary to dissect levels I and IIB in hypopharyngeal cancer?

CONCLUSION: The low incidence of metastases in levels I and IIB in patients with hypopharyngeal cancer in cases of clinical N0 and N+ neck and the fact that all patients with metastases in levels I and IIB received postoperative radiotherapy justifies the preservation of levels I and IIB in patients with hypopharyngeal cancer to improve functional results and reduce the operating time. OBJECTIVES: Neck dissection of levels I and IIB is technically demanding due to the complex local anatomy and can cause several comorbidities. Therefore the aim of the study was to analyze whether levels I and IIB have to be dissected in patients with hypopharyngeal cancer. METHODS: This was a retrospective analysis of all patients who underwent primary surgical treatment for hypopharyngeal cancer and neck dissection, with evaluation of the incidence of metastases in levels I and IIB in cases of cN0 and cN+ neck. RESULTS: None of the patients with cN0 neck but 2/33 patients with cN+ neck had metastases in level I. Metastases in level IIB were detected in 1/14 patients with cN0 neck and 2/36 patients with cN+ neck. All patients with metastases in levels I and IIB received postoperative radiotherapy due to the N2b and N2c status.

Relevance of oropharyngeal cancer lymph node metastases in the submandibular triangle and the posterior triangle apex.

BACKGROUND: Neck dissection of levels I and IIB is time consuming and can cause several comorbidities. The aim was to analyze whether levels I and IIB need to be dissected in patients with oropharyngeal cancer and clinical N0 or N+ neck. PATIENTS AND METHODS: A retrospective analysis of 77 patients with oropharyngeal cancer was carried out with evaluation of the incidence of neck node metastasis in levels I and IIB. RESULTS: None of the patients with cN0 neck had metastases in level I or IIB; 12.8% of the patients with cN+ neck had metastases in level I, 35.1% in level IIA and 25.6% had metastases in level IIB. CONCLUSION: Levels I and IIB should be dissected in cN+ neck in order to achieve maximal oncological safety. The preservation of levels I and IIB in cN0 neck seems to be justified in terms of improving functional results and concomitant reduction of operation time.

Laryngopharyngeal mucosal injury after prolonged use of the ProSeal LMA in a porcine model: a pilot study.

PURPOSE: Little is known about the potential for the cuff of the ProSeal laryngeal mask airway (PLMA) to cause laryngopharyngeal mucosal injury. The aim of this study was to investigate the effects of prolonged exposure of the laryngopharyngeal mucosa to the PLMA in an animal model. METHODS: The PLMA was used for positive pressure ventilation in eight German country pigs randomized to receive different durations of anesthesia lasting three to 24 hr. Two additional pigs served as controls. Biopsies from 14 predetermined areas of the laryngopharyngeal mucosa were examined histopathologically using light microscopy. The number of areas with signs of mucosal damage was evaluated, and the degree of tissue damage was analyzed using a semiquantitative scoring system. RESULTS: The number of areas with moderate to severe mucosal damage was 0-2 in animals anesthetized for less than 12 hr, 6 in animals anesthetized for 12-18 hr, and 12 in animals anesthetized for more than 18 hr. The severity of mucosal damage was also time-dependent, indicated by raising semiquantitative scores for epithelial and subepithelial mucosal damage. CONCLUSION: The histopathological findings of this study show that prolonged use of the PLMA in the pig for up to nine hours is associated with no, or only mild alterations in the laryngopharyngeal mucosa, whereas use for 12 hr or more is associated with clear signs of mucosal injury.