Expression of DOG1 (Using SP31) in Poorly Differentiated Carcinoma of the Head and Neck.

BACKGROUND: The calcium-activated chloride channel protein discovered on gastrointestinal stromal tumour 1 (DOG1) is expressed in a variety of normal and neoplastic tissues. DOG1 is a specific marker for gastrointestinal stromal tumour. In the head and neck region, DOG1 is a sensitive discriminator for acinar cell carcinoma. Only a few publications have presented data concerning the expression of DOG1 in head and neck squamous cell carcinoma (HNSCC). The expression of DOG1 in HNSCC appears to be associated with a poor prognosis. The aim of this study was to analyze the expression pattern of DOG1 in poorly differentiated carcinoma of the upper aerodigestive tract. MATERIALS AND METHODS: A total of 84 specimens from 31 patients with carcinomas of the upper aerodigestive tract were immunohistochemically investigated for DOG1 expression. Inclusion criterion was poorly to undifferentiated carcinoma of the head and neck, but samples of the same resection site that exhibited moderate or well-differentiated squamous cell carcinoma were also enrolled. Immunoreactivity in carcinomas was estimated using a visual score (0: negative; 1: basally positive, 2: parabasally positive, 3: completely positive, 4: basally and parabasally positive). RESULTS: Fifteen out of 84 specimens were immunoreactive to antibody to DOG1 (17.8%). DOG1 immunoreactivity was restricted to eight patients (25.8%). However, DOG1 expression was considerably heterogeneous in tumours, with three (9.6%) cases showing a positive reaction in all samples. Basal and parabasal staining patterns (five specimens each) dominated. DISCUSSION: This study demonstrated expression of DOG1 to be restricted to some poorly differentiated carcinomas of the upper aerodigestive tract. Although the proportion of DOG1-positive carcinomas was moderate compared to results of previous studies on head and neck cancer tissues, DOG1 expression possibly indicates a subset of HNSCC. Further studies are necessary to investigate the heterogeneity and clinical relevance of DOG1 expression in HNSCC.

Congenital Auricular Malformations: Description of Anomalies and Syndromes.

Half of the malformations in the ear, nose, and throat region affect the ear. Malformations of the external ear (pinna or auricle with external auditory canal [EAC]) are collectively termed microtia. Microtia is a congenital anomaly that ranges in severity from mild structural abnormalities to complete absence of the external ear (anotia). Microtia occurs more frequently in males (∼2 or 3:1), is predominantly unilateral (∼70-90%), and more often involves the right ear (∼60%). The reported prevalence varies geographically from 0.83 to 17.4 per 10,000 births. Microtia may be genetic (with family history, spontaneous mutations) or acquired. Malformations of the external ear can also involve the middle ear and/or inner ear. Microtia may be an isolated birth defect, but associated anomalies or syndromes are described in 20 to 60% of cases, depending on study design. These generally fit within the oculo-auriculo-vertebral spectrum; defects are located most frequently in the facial skeleton, facial soft tissues, heart, and vertebral column, or comprise a syndrome (e.g., Treacher Collins syndrome). Diagnostic investigation of microtia includes clinical examination, audiologic testing, genetic analysis and, especially in higher grade malformations with EAC deformities, computed tomography (CT) or cone-beam CT for the planning of surgery and rehabilitation procedures, including implantation of hearing aids.

Expression of podoplanin in primary and metastatic poorly differentiated and undifferentiated carcinomas of the head and neck.

UNLABELLED: The invasive growth properties of head and neck carcinoma can be determined by proteins associated with cellular motility. Podoplanin is a recently identified protein associated with motility of cells. MATERIALS AND METHODS: Sixty-five tumour specimens of 51 patients with poorly and undifferentiated carcinomas were investigated for podoplanin expression. All tissues were fixed in buffered formalin and embedded in paraffin. RESULTS: Podoplanin expression showed a tendency towards a more intense staining in carcinomas with a squamous epithelia differentiation compared to tumours without any remnants of cellular layers. Podoplanin expression was very rarely seen in lymph node metastases and the staining was weak in these specimens. The differences of podoplanin expression between primary and metastatic carcinoma were highly significant (p<0.005). CONCLUSION: Podoplanin is expressed in primary undifferentiated carcinoma, both inside and outside of Waldeyer's ring. This study shows that podoplanin can be used as a marker of tumour invasion in poorly or undifferentiated head and neck carcinoma.

Immunohistochemical study on an epithelial-myoepithelial intercalated duct carcinoma transplanted to the nude mouse.

Parotid gland carcinomas are rare. Among them, the epithelial-myoepithelial carcinoma (EMC) is extremely rarely diagnosed. This case report was based on a female with a history of 17 years of recurrent EMC of the parotid and evidence of distant metastasis over a period of 2 years. During debulking procedures of her extensive facial tumour, small tumour samples were transplanted to nude mice. The tumours grew well on the mice and were characterized morphologically and immunohistochemically after explantation. Cellularity per mm(2) ranged between 3,470 and 7,410. The tumours were characterized by the typical bipolar pattern of tumour cells and broad stromal septae. All but one of 7 transplanted tumours were positive for pan-cytokeratin marker KL-1. The proliferation index in terms of MIB-1-stained nuclei increased from 2% to 20% and was correlated positively to the expression of EGFR. IGF-1R-, VEGF- and FLK1-stained cells were found in all cases. The increase in EGFR- and MIB-1-positive cells correlated with the clinical course of the patient, who showed shorter periods of tumour recurrence prior to her death. These findings in EMC transplanted to the nude mouse demonstrate the feasibility of growing EMC in vivo.

Expression of neuron-specific enolase in irradiated salivary glands of the rat: a pilot study.

External irradiation of advanced head and neck cancer often includes the major salivary glands, despite them not being affected by the disease. The aim of this study was to determine neuron-specific enolase (NSE) as a marker of inflammation in irradiated and non-irradiated salivary glands of the rat. This study shows that NSE expression in irradiated salivary glands is dependent on the topography of the organ related to the irradiation source. NSE immunostaining was increased in irradiated rats, both in glands directly exposed to the irradiation source and in shielded glands, and compared to glands of non-irradiated animals. Staining of NSE in salivary glands appears to indicate the inflammatory response of the organ to X-ray exposure. The increased immunoreactivity for NSE in shielded glands might be useful as a marker of irradiation in salivary glands, even for scattered radiation effects in major salivary glands.

Morphometrics of the facial nerve following fractionated external irradiation: an experimental study in rats.

UNLABELLED: This study provides morphometric data on radiation-induced alterations of the facial nerve in female Wistar rats. The facial nerves were explanted 3 to 4 or 7 to 9 months after completion of a fractionated external irradiation of the left side of the neck (5 days per week, 6 weeks, total: 60 Gray). Both facial nerves were investigated in order to identify possible effects of scattered irradiation on them. A total of 54 animals were investigated and 11,193 measurements were obtained. The computer-assisted image analysing system CUE-3 Color Image Analyzer (Olympus, Japan) was used. Facial nerves from untreated animals served as controls. RESULTS: Three to four months after irradiation, the ratio of axon area and total area of the cross-sectioned nerve was shifted towards the axon area. Seven to 9 months after completion of irradiation, both the myelin sheath and the axon were reduced in diameter. The loss of substance affected predominantly the myelin sheath. Peripheral nerves are radiosensitive. Seven to 9 months after irradiation there was an increase of the axon area compared to the total area, both in absolute values and in relative terms. Radiosensitivity of peripheral nerves can be measured morphometrically in suitable animal models. The measurements disclosed temporal and spatial patterns of radiation response of the nerves. These results may partly explain the increased radiosensitivity of the peripheral nervous system observed in the long-term follow-up after completion of radiotherapy of head and neck in humans.

Insulin-like growth factor-1 receptor (IGF-1R) in primary and metastatic undifferentiated carcinoma of the head and neck: a possible target of immunotherapy.

BACKGROUND: Insulin-like growth-factor 1 receptor (IGF-1R) plays a key role in the development of cancer and is important for the regulation of tissue growth, predominantly via the growth hormone (GH). Targeting IGF-1R is a current topic of cancer research. The aim of this study was to identify IGF-1R in undifferentiated carcinomas of the head and neck. PATIENTS AND METHODS: Twenty-one formalin-fixed and paraffin-embedded specimens from 20 patients with undifferentiated carcinomas were investigated. IGF-1R was detected immunohistochemically. In some cases, the infection status for Einstein Barr virus (EBV, DNA) or HPV (RNA) was known. RESULTS: IGF-1R was detected in all tumours, regardless of differentiation or proof of EBV or HPV integration into the genome. DISCUSSION: This study revealed a broad expression of IGF-1R in undifferentiated carcinomas of the oropharyngeal and nasopharyngeal region, both in primaries and regional lymph nodes. These results suggest that IGF-1R expression in these tumours is capable of transmitting mitogenic signals to the neoplastic cells. The presence of IGF-1R in undifferentiated carcinomas suggests that these tumours can respond to IGF. Targeting this receptor may be a promising tool for the treatment of advanced stage NPC.

Vascular endothelial growth factor (VEGF) in sera of oral and oropharyngeal squamous cell carcinoma patients.

This study was performed to investigate the serum levels of vascular endothelial growth factor (VEGF) in patients with oral and oropharyngeal cancer. The study population included 86 patients with untreated cancer of this region. VEGF serum levels varied considerably and showed no correlation to tumour size or locoregional spread. The wide variety of VEGF serum levels make this marker difficult to handle for initial diagnostics or monitoring of therapy.

Development of fMRI tests for children with central auditory processing disorders.

The term 'central auditory processing disorder' (CAPD; in German-speaking countries: 'auditive Verarbeitungs- und Wahrnehmungsstörung', AVWS) embraces various complex disorders of central auditory function. The purpose of this preliminary study was to develop a functional magnetic resonance imaging (fMRI) test set in order to extend the diagnostic work-up to include imaging, and thus to improve both diagnostic performance and the monitoring of treatment outcome. fMRI was performed in 11 healthy children ranging in age from 7 to 10 years and in 11 healthy adults (age range: 23 to 31 years). Three tests, used for diagnosis of CAPD, served as the basis for fMRI paradigms: the Hannover phoneme discrimination test (HPDT); the auditory memory span test (MST) and the dichotic listening test (DLT). In the HPDT, activations in the dorsal part of the superior temporal gyrus (STG) were found on both sides, in the Broca area and in the left middle temporal gyrus, typical for phonemic tasks. The MST showed bilateral activations of the STG, typical for processing of pseudo-words. In addition, bilateral activation of the hippocampus was found. However, there was no clear activity in the left supramarginal gyrus, where the phonological store is thought to be located. The DLT showed activations of the bilateral STG and of the left inferior frontal gyrus. Predominantly, the paradigms showed typical activation patterns of the examined central auditory functions. These results can serve as a reference for future examinations in children with AVWS/CAPD.

Rapidly progressive and metastatic mucoepidermoid carcinoma: application of serological tumor markers.

Mucoepidermoid carcinoma (MEC) of the salivary gland is a rare entity. A distinction of 2 variants has been proposed: the low-grade tumor with a favourable prognosis and the high-grade tumor with a poor prognosis. Indeed, MEC is a cancer with a relative favourable outcome and more than 90% of patients survive for more than 5 years after diagnosis, reduced to about 70% after 10 years. This excellent prognosis might contribute to the unacceptable retention of the term "mucoepidermoid tumor" in the medical terminology, even in current medical textbooks. However, the distinction of MEC by grading is a guideline only and it is not appropriate to use this histological term as a prediction for individual cases. We describe the rapid fatal outcome of a patient with MEC in order to emphasize the malignant characteristics of this tumor and the possible application of tumor markers for the diagnosis of metastasizing MEC.

Classification and diagnosis of ear malformations.

In the ENT region 50% of the malformations affect the ear. Malformations of the outer and middle ear are predominantly unilateral (ca. 70-90%) and mostly involve the right ear. Inner ear malformations can be unilateral or bilateral. The incidence of ear malformations is approximately 1 in 3800 newborns. Ear malformations may be genetic (associated with syndromes or not, with family history, spontaneous mutations) or acquired in nature. Malformations can affect the outer ear (pinna and external auditory canal, EAC), middle ear and inner ear, not infrequently in combination. Formal classification is advisable in order to be able to predict the prognosis and compare treatment schedules. Various classifications have been proposed: pinna and EAC malformations according to Weerda [1], middle ear malformations according to Kösling [2], and inner ear malformations according to Jackler [3], [4], to Marangos [5] and to Sennaroglu [6]. Additionally, we describe Altmann's classification of atresia auris congenita [7] and the Siegert-Mayer-Weerda score [8] for EAC and middle ear malformations, systems of great practicability that are in widespread clinical use. The diagnostic steps include clinical examination, audiological testing, genetic analysis and, especially, CT and MRI. These imaging methods are most usefully employed in combination. Precise description of the malformations by means of CT and MRI is indispensable for the planning and successful outcome of operative ear reconstruction and rehabilitation procedures, including cochlear implantation.

Duplication of Xq26.2-q27.1, including SOX3, in a mother and daughter with short stature and dyslalia.

Duplications of the distal long arm of the X chromosome are rare and carrier females are usually phenotypically normal. We report on a 14-year-old short statured (height and weight <3rd centile) girl with dup(X)(q26.2q27.1) inherited from a short mother. The proband has minor dysmorphic features, lordosis, lack of menarche, late signs of puberty, low prepuberal levels of gonadotrophins and steroids, but borderline low IGF-1 and normal IGF-Bp3 serum levels. Both the proposita and her mother have severe speech problems with stuttering and dyslalia. The 44-year-old mother with a strikingly aged face and a prominent nose, had menarche at 15 years. Both maternal sisters and the grandmother of the proposita are also short. Karyotyping revealed an additional band at Xq26 in all metaphases from the proband, her mother, and two maternal aunts. Molecular cytogenetic investigations revealed an Xq26.2-q27.1 direct duplication of approximately 7.5 Mb that encompasses or disrupts the SOX3 gene, which maps at the distal border of the duplicated segment. A similar chromosomal duplication was reported recently in five families and in each was associated with an abnormal phenotype in males with short stature [Hol et al., 2000; Solomon et al., 2002, 2004]. Using an androgen-receptor (HUMARA) gene methylation assay and FISH, we show that despite preferential inactivation of the dup(Xq) chromosome a significant proportion of lymphocytes in both mother and daughter carry an active duplicated X chromosome. Our findings further suggest that a dosage effect of SOX3 may to be responsible for a speech disorder in addition to short stature secondary to hypopituitarism.

Epstein-Barr virus DNA and epithelial markers in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) belongs to the most common malignant tumours in certain parts of the world, e.g. South-East Asia. The undifferentiated type of NPC is associated with genomic Epstein-Barr virus (EBV) DNA. In normal epithelia of the nasopharynx cytokeratins (CK) 4, 5, 6, 13, 14, 15 and 19 are expressed. The aim of this study was to analyse the expression pattern of cytokeratins in NPC in the presence of EBV infection. Twenty primary or metastatic tumours from 13 patients suffering from a NPC were evaluated (formalin-fixed, paraffin-embedded). (35)S-labelled probes were used to detect EBV DNA in the tissue sections. Fourteen specimens (70%) were EBV positive. All positive specimens were undifferentiated NPC. All NPC were identified with broad-spectrum anti-CK antibody. Using a panel of anti-CK antibodies, there was no specific CK-expression pattern in NPC. In summary, undifferentiated NPC are strongly associated with EBV. The cytoskeleton of undifferentiated NPC reveals no specific pattern of CK expression.

The effect of external fractionated irradiation on the distribution pattern of extracellular matrix proteins in submandibular salivary glands of the rat.

INTRODUCTION: The aim of this study was to analyse the distribution pattern of extracellular matrix proteins in the irradiated and non-irradiated rat submandibular salivary gland in order to provide a more detailed profile of the radiation injury following radiotherapy of the head and neck. MATERIAL AND METHODS: External X-ray exposure, restricted to the left skull base and neck region, was performed in 60 female Wistar rats, fractionated to daily applications of 2 Gy, up to total dosages of 20, 40 or 60 Gy. Both submandibular glands were excised after supravital anaesthesia 6 months or 1 year after completion of the irradiation. Spatial and temporal patterns of extracellular matrix proteins were investigated histologically and immunohistochemically. RESULTS: The polyclonal anti-human antisera used, identified the same antigens in rat tissue as in human tissues. The alterations in staining patterns and staining intensities between irradiated and non-irradiated salivary glands showed statistically significant differences. Different structures in irradiated glands reacted with different intensities, e.g. nerve tissue and the basement membranes of excretory ducts were intensely laminin-positive, fibronectin was predominantly found around the excretory ducts with transition to the interstitial tissues. CONCLUSION: Irradiation leads to statistically significant differences in the amount and composition of the extracellular matrix in salivary glands. The amount of extracellular matrix proteins in irradiated glands is dose-dependent. The higher the dosage the more extracellular matrix can be expected. Consecutively, total dosage is associated with greater loss of acini. Scatter effects of irradiation have also to be recognized. Immunohistochemical studies on salivary glands have to consider the pretreatment status, in particular those studies that investigate degenerative changes.