Members of the lysyl oxidase family (LOX) are copper and lysyl‐tyrosine quinone cofactor‐containing amine oxidases that are important for the assembly and maintenance of components of the extracellular matrix. Our previous results demonstrated that a novel member, LOXL4, is overexpressed in head and neck squamous cell carcinoma (HNSCC) compared to normal squamous epithelium. Results of the current study showed overexpression of the LOXL4 transcript in 74% (46 of 62) of invasive HNSCC tumours and 90% of both primary and metastatic HNSCC cell lines. Significant correlation was found between LOXL4 expression and local lymph node metastases versus primary tumour types (p < 0.01) and higher tumour stages (p < 0.01). Immunocytochemistry demonstrated cellular overexpression of the LOXL4 protein that correlated with the increased mRNA transcription in HNSCC cells. HNSCC cell lines displayed in significant subset of nuclei increased copies of the LOX4 gene locus on chromosome 10q24, demonstrated by fluorescence in situ hybridization (FISH). Extensive metaphase cytogenetic analysis was performed on UTSCC19A cells, identifying an isochromosome i(10)(q10). Taken together, these results highlight LOXL4 expression as a distinctive trait and suggest a functional role for LOXL4 in the molecular pathogenesis of invasive head and neck carcinomas. Copyright © 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Die vorliegende Kasuistik beschreibt den Fall eines 67-jährigen Patienten mit plötzlicher Atemwegsobstruktion durch spontane Weichteileinblutungen in die Zunge, Mundhöhle und den Hypopharynx ohne anamnestisch bekannte hämorrhagische Diathese. Bei Erstkonsultation bestanden zusätzlich zu einem Globusgefühl mit Dysphagie eine kloßige Sprache und eingeschränkte Mundöffnung. Die Computertomographie zeigte weichteildichte Raumforderungen von Mundboden, Zungengrund und rechter Pharynxseitenwand. Am folgenden Tag musste der Patient wegen plötzlicher respiratorischer Insuffizienz aufgrund einer kompletten Verlegung der Atemwege bei nicht durchführbarer Intubation koniotomiert werden. Die Gerinnungsdiagnostik zeigte eine herabgesetzte Faktor-VIII-Aktivität durch den Nachweis von gegen Faktor VIII gerichteten Autoantikörpern. Anhand dieser Kasuistik und einer vergleichenden Übersicht über die Literatur stellen wir die Diagnostik und Therapie der erworbenen Hämophilie A dar.

OBJECTIVE:To describe the outcome of cochlear implantation in children and to discuss the cause and management of cochlear reimplantation.STUDY DESIGN:Retrospective chart review.METHODS:The medical records of 110 patients younger than 18 years of age, who underwent cochlear implantation at the Department of ORL, Head and Neck Surgery, of the University of Kiel, Germany, were reviewed for demographics, complications, and history of revision surgery. The patients had previously had implantation with either Nucleus (including the Contour) devices or MED-EL devices.RESULTS:Length of use before cochlear explanation ranged from 4 days to 3.9 years. Reimplantation was caused by traumatic device failure, wrong electrode insertion and infection of implanted area. Results indicated a reimplantation rate of 5.4% in children compared to 0.8% in adults, mostly resulting from the greater risk of children receiving an impact to the head. Postoperative performance data showed no decrease in scores taken before failure.CONCLUSIONS:Though young children who are developing their motor skills are probably at greater risk of a cochlear reimplantation resulting from device failure following head trauma, surgical revision with reimplantation can be performed safely and without decrement to performance.

We report the case of a 67 year old patient suffering from acute airway obstruction caused by hemorrhage localized to the tongue, mouth cavity and hypopharynx, with no evidence of bleeding in his history. The patient presented initially with a globus feeling of the neck, dysphagia and a sore throat. CT scan revealed a swelling of the lingual and sublingual areas and the pharyngeal wall. Next day, there was an immediate life-threatening event caused by progressive bleeding with airway obstruction and an inability to intubate requiring coniotomy. The etiology of the hemorrhage was confirmed by finding a depletion of factor VIII and the presence of auto-antibody directed against this factor. Based on this case report and a review of the literature, we discuss the diagnosis and treatment of acquired hemophilia.

Für eine Immuntherapie von Tumorerkrankungen scheinen DC-basierte Vakzinierungsansätze vielversprechend, da DCs endogen und exogen aufgenommene Antigene den körpereigenen T-Lymphozyten in Assoziation mit MHC-Klasse I und II Molekülen präsentieren können, über verschiedene kostimulatorische Signale für die T-Zellaktivierung (B7-1, B72) verfügen und wichtige Adhäsionsmoleküle exprimieren. Die Gewinnung von DCs aus menschlichen Blutmonozyten und die Präparation nach diesem hier vorgestellten Protokoll ermöglicht die Verwendung der DC als Vakzine für Immuntherapien von Kopfund Halskarzinomen (Abb. 1). Herstellung dendritischer Zellen (DC) aus menschlichen peripheren Blutzellen zur Immuntherapie von Patienten mit Kopfund Halskarzinomen

We report the case of a 67 year old patient suffering from acute airway obstruction caused by hemorrhage localized to the tongue, mouth cavity and hypopharynx, with no evidence of bleeding in his history. The patient presented initially with a globus feeling of the neck, clysphagia and a sore throat. CT scan revealed a swelling of the lingual and sublingual areas and the pharyngeal wall. Next day, there was an immediate life-threatening event caused by progressive bleeding with airway obstruction and an inability to intubate requiring coniotomy. The etiology of the hemorrhage was confirmed by finding a depletion of factor VIII and the presence of autoantibody directed against this factor. Based on this case report and a review of the literature, we discuss the diagnosis and treatment of acquired hemophilia.

Candida albicans (CA) is a frequent opportunistic pathogen in cancer patients. Usually, human surfaces are protected, apart from physical barriers, by the production of human beta-defensins (hBD). hBD-2 shows a potent antimicrobial activity against CA. We therefore investigated whether CA induces hBD-2 expression in primary oral cells and if immunosuppressive betamethasone alters hBD-2 expression. Additionally, we studied, whether a lack of hBD-2 expression could explain opportunistic infection of tonsillar cancer. Primary oral epithelial cells and fibroblasts were stimulated with Candida albicans in a time- and dose-dependent manner with or without betamethasone preincubation. Total RNA from oral cells and specimens was isolated and hBD-2 expression was analyzed by semiquantitative RT-PCR. Our data demonstrate that opportunistic CA induced hBD-2 expression in a time- and dose-dependent manner, suggesting hBD-2 to be a fast antifungal, epithelia-derived immune response. Treatment with glucocorticoid could lead to diminished innate immunity based on suppression of inducible AP. Malignant transformation induces alteration of hBD-2 expression and leads to a reduced hBD-2 expression and subsequentially to Candida colonization on oral SCCs.

Die Vakzinierung von Krebspatienten mit autologen tumorantigen-präsentierenden dendritischen Zellen (DC) kann eine Immunantwort gegen das Tumorgewebe auslösen und eine anhaltende Immunität erzeugen, wie verschiedene Studien gezeigt haben. Wir stellen ein Konzept zur Immuntherapie vor, das Antigen in Form von RNA nutzt und mit transfizierten DC eine T-Zellantwort auslösen soll. Zur Präparation von reifen DC wurden periphere Blutleukozyten durch Leukapherese gewonnen und mit verschiedenen Zytokinen für 7 Tage kultiviert. Bei Plattenepithelkarzinomen des Kopf-Halses (HNSCC) ist das Gen LOXL-4 zu 60% überexprimiert und für ein Vakzinierungskonzept geeignet. Die gesamte kodierende Region von LOXL-4 wurde zunächst in einem Klonierungsvektor (pSP73/sph/A64) eingebaut, in kompetente Zellen (modifizierte E. coli) integriert und vermehrt. Anschließend wurde das Plasmid wieder extrahiert und durch Restriktionsanalyse die Insert-DNA kontrolliert. Nach Sequenzierung des geklonten DNA-Fragmentes und reverser Transkription wurden reife DC mit LOXL-4-mRNA durch Elektorporation transfiziert, die Transfektionsrate anschließend mit anti-LOXL-4-Ak bestimmt. Die Ergebnisse zeigen, dass die Herstellung einer Vakzine auf Basis von RNA transfizierten DC zur adjuvanten Immuntherapie von Malignomen des Kopf und Halses möglich ist. Die Expression des Gens auf den DC erlaubt nun weitere Untersuchungen zum Nachweis der immunologischen Fähigkeit der Vakzine, u.a. der Induktion von zytotoxischen T-Lymphozyten.

BACKGROUND:Hybrid cells generated from dendritic cells (DC) and tumour cells provide tumour-associated antigens (TAA) in a polyvalent mode. The present study was designed to investigate the hybrid cell generation by dendritic cells and different tumour cell lines to establish an electrofusion protocol with an optimal fusion setting.MATERIALS AND METHODS:Hybrid cells from mature DC and tumour cells were generated by electrofusion. Fusion efficiency was determined by flow cytometry, as well as by light and fluorescence microscopy analyses.RESULTS:The gradual electrofusion process constituted different human dendritic cell tumour cell hybrids of high diversity depending on electrical and non-electrical parameters. Factors influencing fusion frequency were determined by specific cell staining with mAbs, FACS analysis and trypan blue dye exclusion.CONCLUSION:Increased fusion efficiency was associated with reduced viability. The protocol presented in this work might be helpful for future fusion studies as a prerequisite for comparable in vitro and human vaccination trials.

The palatine tonsils have an undoubted role in the immune defence system. After antigen contact an effective adaptive immune response by B- and T-cell lymphocytes will be released. In addition the palatine tonsils seem to exert influence to the defence by the innate immune system. Therefore, we studied the ability of palatine tonsils to express different alpha and beta defensins and to find out any distinctions in chronic inflamed tonsils. Total RNA of 49 specimens of hyperplastic tonsils and chronic tonsillitis with pathological provided evidence of Actinomyces israelii was isolated using TRIzol protocol, reverse transcribed and the HNP-1, HNP-4, HBD-1 and HBD-2 gene expression densitometric determined, standardised in relation to glycerinaldehyd-3-phosphatdehydrogenase gene expression, after a semiquantitative polymerase chain reaction was performed. mRNA of HNP-1, HNP-4, HBD-1 and HBD-2 was detected in tissue samples, but their amount differed within the two defensin families and tissue of origins. HBD-1 was detected in all 49 tissues of hyperplastic tonsils and chronic tonsillitis. Only in chronic inflamed tonsils the amount of HBD-2 mRNA expression was significant increased. In these specimens also mean relative expression rate of all defensins was observed to be manifestly increased. Palatine tonsils express mRNA for different alpha and beta defensins and this expression suggest a newly supposed function in immune defence: the participation in the innate, non-adaptive immune system. Thus, palatine tonsils have a potentially influence in the growth and control of the physiological mouth bacteria by their bactericidal activity.

Chemokines in biological sample are frequently found at very low level. Further, concentrations of chemokines which were measured in the same tissue detected with different methods often differ in literature. Therefore, RANTES concentrations were quantified by application of sandwich enzyme-linked immunosorbent assay (ELISA) and Western blot analyses. The sensitivity of ELISA was found to be much higher than that of Western blot. Additionally, the detection time differed considerably as well. For biological and biochemical characterization of chemokines it is essential to implement the optimal purification and detection techniques. With an understanding of the technical procedures and some pitfalls, chemokine detection can be applied more reliably.