Interstitial hypertension in head and neck tumors in patients: correlation with tumor size.

Elevated interstitial fluid pressure (IFP) is associated with poor blood supply and inadequate delivery of drugs to solid tumors. IFP was measured in squamous cell carcinomas of the head and neck region in humans using the wick-in-needle technique. In all lesions (n = 19), the IFP was elevated (4-33 mm Hg). Furthermore, the IFP increased with tumor size. The highest IFP was 33 mm Hg in a 24-ml tumor. In one tumor, the IFP was found to be negative (-2.6 mm Hg), which is comparable to that in human skin or subcutaneous tissue. The histopathology of this tumor was benign. If this pressure difference between malignant and benign lesions can be confirmed in a large number of tumors, then the IFP could be used to aid tumor detection during needle biopsy. The value of IFP as a predictor of response to radiotherapy, photodynamic therapy, hyperthermia, and chemotherapy should be assessed prospectively.

Phthalates demonstrate genotoxicity on human mucosa of the upper aerodigestive tract.

Various phthalate compounds are used as softeners and plasticizers in a wide range of plastic materials. There has been a growing concern regarding a possible health hazard to humans. The mucosa of the upper aerodigestive tract is the organ of first contact for the majority of xenobiotics, such as phthalates, entering the body. Still, there is a lack of information concerning possible carcinogenicity of phthalates in the upper aerodigestive tract. This motivated us to investigate their genotoxic effects on human epithelia: human mucosal cells derived from biopsies harvested during surgery of the oropharynx and the inferior nasal turbinate, respectively. The alkaline version of the microgel electrophoresis assay was used to detect single-strand breaks in the DNA following incubation with dibutylphthalate (DBP) and diisobutylphthalate (DiBP). DNA damage was induced by both DBP and DiBP in oropharyngeal and nasal mucosa, though the effect of DiBP was more pronounced than that of DBP. Nasal mucosa proved to be more sensitive than oropharyngeal epithelia. The results demonstrate genotoxic effects of phthalates on human mucosal cells of the upper aerodigestive tract, in contrast to earlier findings in animal models.

A special method of intra-arterial infusion for treatment of head and neck cancer.

Immediately after radical neck dissection the external carotid artery is lengthened by an autogenic saphenous vein graft and sutured more proximally to the common carotid artery. All branches of the external carotid artery not contributing to the blood supply in the tumor region are ligated. After the wound has healed, the vascular graft will be easily palpable and can be punctured repeatedly for highly selective intra-arterial chemotherapy of the inoperable primary tumor for a prolonged period of time. The method seems to be practicable, innocuous, and more effective than conventional methods of intra-arterial chemotherapy for head and neck cancer.

Ep-CAM--a marker for the detection of disseminated tumor cells in patients suffering from SCCHN.

Disseminated tumor cells are considered as the origin of metastases. Since the number of circulating tumor cells in the bone marrow or the peripheral blood of patients correlates well with the tumor stage, their early detection is an important feature for the identification of high risk patients. We therefore investigated the pan-carcinoma antigen Ep-CAM for its suitability to serve as a specific marker for disseminated tumor cells in patient with squamous cell carcinoma of the head and neck (SCCHN). In order to detect small numbers of tumor cells in early tumor stages, we developed and describe here a RT-PCR assay that detects a single tumor cell within 10(5) normal cells. We examined bone marrows from patients and healthy donors and demonstrate that Ep-CAM can be used as a tumor marker for the diagnosis of single tumor cells in patients with SCCHN.

Targeting head and neck cancer by GM-CSF-mediated gene therapy in vitro.

BACKGROUND: The prognosis for patients with squamous cell carcinoma of the head and neck (SCCHN) has remained poor during the last decades, emphasizing the need for new treatment modalities. Consequently, the objective of our study was to evaluate the feasibility and efficacy of cytokine-mediated gene therapy in SCCHN in vitro. MATERIALS AND METHODS/RESULTS: The SCCHN cell line PCI-1 was transduced by lipofection with a plasmid encoding the human granulocytemacrophage colony-stimulating factor (GM-CSF). Transfection of PCI-1 resulted in the production of significant amounts of GM-CSF as tested by ELISA. Enhanced proliferation of a GM-CSF sensitive cell line, TF-1, after incubation with supernatants of GM-CSF-transduced tumor cells demonstrated the release of biological active GM-CSF from these PCI-1 cells. In addition, GM-CSF-secreting PCI-1 cells enhanced antitumor cytotoxicity of allogeneic peripheral blood mononuclear cells, as tested in 24h-MTT-cytotoxicity assays. CONCLUSIONS: Our data demonstrate the feasibility and efficacy of GM-CSF-mediated stimulation of the antitumor immune response against SCCHN in vitro and may help to define new strategies in the treatment of this malignancy.

Mutagen sensitivity of nasopharyngeal cancer patients.

Primary nasopharyngeal carcinomas (NPCs) may be of various types, including squamous cell carcinomas, undifferentiated carcinomas, and lymphoepitheliomas. Tumor initiation has been linked to the Epstein-Barr virus and, in some geographical regions, to alimentary factors. Possible hereditary components for the appearance of NPCs have not yet been clearly identified. In this study, genetic sensitivity to the genotoxic effects of carcinogenic xenobiotics as an endogenous risk factor of tumor initiation was investigated. The single cell microgel electrophoresis assay was used to quantify chemically-induced DNA damage in lymphocytes of 30 NPC patients and 30 non-tumor donors. The xenobiotics investigated were N'-nitrosodiethylamine, sodium dichromate, and nickel sulphate, with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and dimethyl sulfoxide (DMSO) as positive and negative controls, respectively. The extent of DNA migration in the solvent control cultures was not significantly different between the two groups (1.2+/-0.5 mean Olive tail moment and standard deviation of 30 individuals for NPC patients; 1.1+/-0.4 for non-tumor donors). With constant exposure and electrophoretic conditions, genotoxic effects of varying degrees were induced by the different xenobiotics in tumor and non-tumor patients (nickel sulphate: 7.1+/-2.5 for NPC patients and 5.9+/-1.6 for non-tumor donors; sodium dichromate: 18.1+/-5.3 for NPC patients and 16.2+/-5.4 for non-tumor donors; MNNG: 47.8+/-13.3 for NPC patients and 52.7+/-13.6 for non-tumor donors). Only N'-nitrosodiethylamine proved to induce significantly more DNA migration in lymphocytes of tumor patients (9.8+/-3.1) as compared to non-tumor patients (8.2+/-2.3). Although for sodium dichromate the degree of DNA migration did not significantly differ, variability in migration patterns proved to be lower in the tumor group. Mutagen sensitivity of NPC patients was shown to be elevated for a selected xenobiotic, whereas a general elevation of DNA fragility was not present. Further studies on mutagen sensitivity as an endogenous risk factor influencing the susceptibility of patients at the time of first diagnosis of nasopharyngeal carcinomas are warranted.

Comparing the genotoxic sensitivities of human peripheral blood lymphocytes and mucosa cells of the upper aerodigestive tract using the Comet assay.

Carcinogenesis in the upper aerodigestive tract is influenced by multiple factors. Besides tobacco and alcohol consumption, specific pollutants such as phthalates, nitrosamines, and polycyclic aromatic carbohydrates may be important in tumor initiation. Genetic factors related to mutagen sensitivity and DNA repair capacity also play a role. The aim of this study was to investigate whether human peripheral blood lymphocytes and mucosal epithelium of the upper aerodigestive tract, the target for volatile and liquid xenobiotics, are equally sensitive to genotoxic agents. The Comet assay was used to detect for DNA damage induced by genotoxic agents in mucosal epithelial cells and peripheral blood lymphocytes of 60 volunteers. Mucosa was harvested from larynx, oropharynx, and inferior nasal turbinates. Xenobiotics investigated were dibutylphthalate (DBP), diisobutylphthalate (DiBP), N'-nitrosodiethylamine (NDELA), benzo[a]pyrene (B[a]P), and N'-methyl-N'-nitro-N-nitrosoguanidine (MNNG). DBP, DiBP, B[a]P, NDELA and MNNG induced a significant increase in DNA migration in both cell populations. Peripheral blood lymphocytes were more sensitive than mucosal cells to DBP and DiBP, but not to NDELA and B[a]P. The correlation, in terms of DNA migration, between lymphocytes and mucosal cells among volunteers was relatively poor. Based on the poor correlation in response between the two cell types, the sensitivity of peripheral blood lymphocytes to genotoxic agents appears to be a poor predictor of sensitivity in the target cells of the upper aerodigestive tract. Further attention should be focused on intra-individual mutagen sensitivities and inter-individual genetic differences as regards susceptibility to upper aerodigestive tract cancer.

A comparison of class II antigenicity of human tracheal allografts stored in cialit and in merthiolate.

This report deals with some of the immunological aspects of the transplantation and preservation of human tracheal grafts. The immunological behavior of the transplanted graft depends largely on the interaction of the preservatives with the tissue proteins. Using monoclonal antibodies and the immunoperoxidase staining technique we have investigated the effect of Merthiolate, Cialit, formaldehyde/Merthiolate, and formaldehyde/Cialit preservation techniques on monomorphic determinants of class II transplantation antigens in human tracheal allografts. Unpreserved grafts were found to express class II antigens. These antigens were totally destroyed after 7 days in formaldehyde and 42 days in Cialit and Merthiolate. Preservation in Cialit and Merthiolate showed a gradual disintegration of the histological structures. In contrast, buffered formaldehyde did not appear to alter the histological structure of the tracheal graft. Irrespective of the mode of preservation, the cartilaginous tissue appeared to persist virtually unaffected. No essential differences were observed between the immunological staining of tracheal grafts preserved in Cialit and Merthiolate.

Influence of chemical allograft preservation procedures on the human immunodeficiency virus.

Since chemically preserved allogenic transplants have an established place in reconstructive procedures, the possibility of transferring the human immunodeficiency virus (HIV) with these transplants has been intensively discussed. In this study the authors obtained brain and spleen samples from six HIV-infected cadavers and preserved them with Merthiolate, Cialit, and formaldehyde. After preservation, the tissues were examined for proviral HIV-1 DNA (gag, pol, env) using the polymerase chain reaction. Proviral sequences were clearly demonstrated after the preservation procedure. The results of this study indicate that HIV remains in tissues that have been treated with Merthiolate, formaldehyde, or Cialit. Further investigations are necessary to determine if the virus is in an inactivated or activated form. It can be concluded that, because of the possible transmission of HIV by chemically preserved homografts, serologic screening of donors should be mandatory.

Basic innervation pattern and distribution of classic autonomic neurotransmitters in human nasal mucosal vasculature.

The neural control of human nasal vasculature is still not completely understood. This study was performed to demonstrate the innervation pattern of the different vessel types and to distinguish between nor-adrenergic and cholinergic structures. General innervation was demonstrated using antibodies to neuron-specific enolase and S-100 protein. Autonomic structures were shown by using antibodies to tyrosine hydroxylase and choline acetyltransferase (ChAT). In addition, choline acetyltransferase (AChe) histochemistry was performed. Nasal vasculature is controlled by a dense innervation that increases with the thickness of the tunica media. While all larger vessels show a mixed autonomic innervation, sympathetic structures seem to predominate in veins. These findings demonstrate that classic neurotransmitters play a major role in the regulation of nasal vasculature. The stronger innervation of arteries and cushion veins underlines their central position in the control of nasal air flow.

Quantitative analysis of interleukin-1-alpha gene expression in middle ear cholesteatoma.

Regardless of its origin, cholesteatoma is characterized by the presence of a keratinizing epithelium with an hyperproliferative behavior leading to a very important bone resorption. Previous studies have demonstrated overexpression of interleukin-1 (IL-1 protein in middle ear cholesteatoma by immunohistochemistry and enzyme-linked immunosorbent assay, suggesting a significant role for IL-1-alpha. In this study, the presence of IL-1-alpha messenger ribonucleic acid (mRNA) was quantified by in situ hybridization on frozen sections (n = 10) and by computer-assisted image analysis. Human skin obtained from the external ear canal (n = 10) was used as the control. A higher percentage of cells hybridized for the antisense probes IL-1-alpha mRNA was found in cholesteatoma epithelium. Furthermore, keratinocytes of the suprabasal cell layers were also found to contain specific hybridizations. Some cells in cholesteatoma stroma also contained IL-1-alpha mRNA transcripts. The results of this study confirm the central role of IL-1-alpha in the epithelium hyperproliferation and bone resorption observed in middle ear cholesteatoma.

Immunobiological peculiarities of cholesteatoma in children: quantification of epithelial proliferation by MIB1.

Cholesteatoma in children is characterized by a more extensive and rapid growth in the middle ear and mastoid cavities. The growth characteristics of the cholesteatoma in 20 children were studied using the monoclonal antibody MIB 1, which recognizes a nuclear antigen expressed by cells in the G1, S, and G2/M phases. Specimens of normal adult auditory meatal skin (n = 15) and adult cholesteatoma (n = 15) served as controls. The tissue specimens were prepared for immunohistochemical examination using the alkaline phosphatase-antialkaline phosphatase method and an automatic image analyzer. Specimens of normal skin revealed an average MIB 1 score of 9.2 +/- 3.10%. Child and adult cholesteatomas showed higher values. The average MIB 1 score was higher in child cholesteatoma (42 +/- 9.4%) than in adult cholesteatoma (28.2 +/- 6%). This difference was statistically significant (P<.01). Our results confirm a significant increase of the proliferative rate of cholesteatoma keratinocytes in children, giving an explanation for the more aggressive clinical behavior observed in these patients.

Relapsing polychondritis: a course over 20 years with cerebral involvement.

Relapsing polychondritis is a chronic inflammatory disease associated with an autoimmune disorder in cartilaginous tissue, eyes, labyrinth, blood vessels, and central nervous system. We describe a 75-year-old woman who presented with a 20-year history of dyspnea, inspiratory stridor, and polyarthritis. She developed dysmorphism of both ears and a saddle nose approximately 10 years earlier. Subsequently, she suffered from hearing loss and a tremor. A T2-weighted magnetic resonance imaging scan of the brain revealed multiple, spotted signal intensities. Immunohistochemical analysis of a serum sample showed antibodies to cartilaginous tissue, which were further identified on immunoblotting as antibodies to type II collagen. The extremely prolonged course of disease (>20 years) until a correct diagnosis was made is remarkable. Also, cerebral involvement, which was most likely caused by cerebral angiitis, and which, to our knowledge, has never previously been reported in this form, was detected. Arch Otolaryngol Head Neck Surg. 2000;126:1495-1498

Monoclonal antibodies against human chondrocytes.

Cell-specific antigens are mainly found in cells or membrane surfaces rather than in the surrounding matrix. However, until now it was not possible to produce antibodies specific for cellular structures of chondrocytes. In 1989, Lance (Immunol. Lett. 21:63-73; 1989) first established specific monoclonal antibodies for human articular chondrocytes tested only by immunofluorescence. Studies describing the specificity of these five antibodies (HUMC 1-5) and their relevance for immunohistological analysis of cartilage tissue were not available until now. Therefore, the aim of the following study was to investigate the distribution of HUMC 1, 2, 3, 4, and 5 in mesenchymal cells in vivo and in vitro immunohistochemically. Further investigations concentrate on the localization of chondrocyte specific antigens using immunoelectron microscopy. Immunohistological studies showed positive immunostainings with all five antibodies in human chondrocytes in vivo and in vitro. A cross-reaction with human fibroblasts and osteoblasts for the antibodies HUMC 2 and HUMC 5 was observed. Furthermore, a parallel loss of immunoreactivity for HUMC 1, HUMC 3, and HUMC 4 was observed in cultured chondrocytes indicating that the specific antigens vanish during differentiation observed in vitro. Subsequent immunoblot analysis employing collagens as antigens did not show any reactivity. Using immunoelectron microscopy, gold particle labeling was observed in intracytoplasmatic vesicles of isolated chondrocytes. Our results indicate that HUMC 1, HUMC 3, and HUMC 4 are specific for cartilage cells and might be suitable for immunohistological analysis of different cartilage tissues and pathologically altered chondrocytes.

Photodynamic therapy in head and neck surgery.

The results of a clinical study of photodynamic therapy (PDT) on early stage cancer in head and neck surgery are presented in this report. 30 patients with T1 malignancies of the face and oropharynx have been treated primarily with PDT. After the longest follow up of 14 months two patients revealed recurrence of the disease or residual tumor, two patients have been retreated because of residual dysplastic cells in the control biopsy, and all other patients stay histologically proven free of disease. Hence, PDT appears to be a promising cancer treatment for different histological tumors that penetrate the host tissue definitely less than a laser beam of 630 nm, which is the appropriate wavelength for PDT. Deeper-penetrating malignomas may be accessable for PDT using insertable filters when applying interstitial laser light.

5-Fluorouracil/folinic acid/cisplatin-combination and simultaneous accelerated split-course radiotherapy in advanced head and neck cancer.

In advanced inoperable head and neck cancer radiotherapy alone is unsatisfying. Better results can be obtained by simultaneous 5-Fluorouracil/Cisplatin-chemotherapy and irradiation. The cytotoxicity of 5-Fluorouracil can be enhanced synergistically by adding Folinic Acid in excess. In a clinical phase II trial 62 previously untreated patients suffering from unresectable AJCC-stage III (4 pts.) and IV (58 pts.) squamous cell carcinoma of the head and neck were treated with a simultaneous chemoradiotherapy consisting of high-dose Folinic Acid in addition to a 5-Fluorouracil/Cisplatin combination and of accelerated split-course radiotherapy. As results, three pts. died from tumor arrosion bleeding during the treatment. Median follow up time of the surviving pts. is 27 + months (range 18-44 months). 48/62 pts. (77%) achieved complete remission, 11/62 pts. (18%) partial remission. Presently, 32 pts. (52%) are without evidence of disease. Actuarial three years overall survival rate (Kaplan-Meier method) out of 62 pts. in 53%. Actuarial disease free survival and local tumor control rates at three years are 58% and 72%. Mucositis was severe but tolerable, bone marrow depression was moderate to marked. In conclusion, this combined simultaneous modality approach is highly effective in locally advanced head and neck cancer. It seems to provide superior survival and local control rates as compared to conventional radiotherapy or sequential chemo-radiotherapy or as compared to simultaneous 5-Fluorouracil/Cisplatin and non-fractionated radiotherapy. A comparative phase III study is required.

Experimental investigations on the suitability of the carbon dioxide laser for stapedotomy.

Surgery of the footplate may cause a number of possible complications such as cochlear hearing defects or even complete loss of hearing. This has made it necessary to look for improved techniques. When a laser is used for stapedotomy, the energy transmitted to the cochlea must be reduced to the lowest possible level. We thus investigated the carbon dioxide laser to determine whether it would prove to be more advantageous than the argon laser. Animal experiments showed that CO2 laser irradiation is well tolerated. Tests performed on isolated petrous bone resulted in the development of a new surgical instrument suitable for stapedotomy using the CO2 laser. Temperature and pressure measurements were carried out on a simplified model of the human cochlea. Our findings indicated that, with both types of laser, the irradiation required to perforate the otosclerotically thickened footplate adversely affects temperature and pressure development in the cochlea.

Functional morphology of nasal blood vessels in humans.

"Secretion" and "obstruction" as predominant clinical symptoms in rhinology affect a great number of patients with disorders of the nose. According to clinical experience as well as morphological investigations, the endonasal vascular system is most likely to be involved in these functional mechanisms. In the present study, we report morphological findings on the angioarchitecture of human nasal mucosa. Meticulous investigation of the structure and ultrastructure of nasal mucosal capillaries revealed differences in the appearance of the endothelial lining. Especially the morphological feature of attenuated fenestrated endothelia in these vessels might be correlated with the functional behaviour under various physiological and pathological conditions. Inspection of the vascular wall of nasal swell bodies revealed differences in the orientation as well as the shape of muscle cells in different parts of this vascular system. The functional role of special morphological features known as muscular bolsters and intervascular muscle fibres for the swelling mechanism of the nasal mucosa is emphasized. Our results enabled us to define the muscular structures that are most probably responsible for constriction and dilatation of nasal swell bodies.

Tracheal transplantation: demonstration of HLA class II subregion gene products on human trachea.

The present morphological study was designed to investigate the expression of HLA class II subregion gene products on human trachea. Frozen sections from the tracheas of 20 cadavers not suffering from ear, nose or throat diseases were studied immunohistochemically using monoclonal antibodies which recognize monomorphic determinants of HLA-DR, -DP and -DQ molecules. Positive reactions could be detected in the airway epithelium and mixed glands. HLA-DR and -DP showed a stronger presence than HLA-DQ. Our results indicate that tracheal mucosa may be the major antigenic structure of the trachea and therefore responsible for the immunogenic action of allogenic tracheal transplants.

The lateral cyst of the neck: congenital or acquired?

An outer rim of lymphoid tissue and an inner epithelial lining of squamous composition form the lateral (branchial) cyst of the neck. According to the particular pattern of keratin polypeptides, branchial mass inner lining is shown to be homologous to upper digestive tract (UDT) squamous epithelia. Furthermore, immunostaining of tissue sections with a polyclonal antibody highly specific for the major acidic UDT keratin 13 demonstrates that the epithelium in question is composed of both basal and normally differentiated, i.e. non-keratinizing suprabasal (spinous) cells. 'Import' into a neck lymph node, and rapid growth of oropharyngeal crypt epithelial cells is suggested to initiate a sequence of events leading to an acquired lymphoepithelial mass that may actually present as a 'branchial cyst'.