Patterns of chromosomal alterations in metastasizing and nonmetastasizing primary head and neck carcinomas.

In an attempt to define chromosomal alterations that are associated with the metastatic phenotype, we investigated a total of 29 metastasizing (pN+) and 19 non-metastasizing (pN0) head and neck squamous cell carcinomas by comparative genomic hybridization (CGH). The analysis indicated that the pN0 tumors carried preferentially overrepresentations of chromosomes 5p, 6p, and 7p and that the pN+ tumors were frequently characterized by deletions on chromosomes 7q, 10q, 11p, 11q, 15q, and 20p and overrepresentations of the chromosomes 19q and 20q. In particular, the use of difference histograms and statistical analysis indicated that the deletions on chromosomes 10q25-q26 and 11p13-p14 were highly significant for metastasizing carcinomas. The findings on chromosome 10q were supported by loss of heterozygosity analysis in the primary tumors and eight synchronous lymph node metastases using four microsatellite polymorphisms. The data suggest that distinct patterns of genetic lesions are responsible for the metastatic phenotype of head and neck squamous cell carcinomas.

Polymorphism within the cyclin D1 gene is associated with prognosis in patients with squamous cell carcinoma of the head and neck.

We have examined the correlation of a frequent A/G polymorphism within exon 4 of the cyclin D1 gene (CCND1) with genetic susceptibility and clinical outcome in 384 patients with squamous cell carcinoma (SCC) of the head and neck. CCND1 genotype frequencies were similar in the cases and 191 controls. Furthermore, the CCND1 genotype was not associated with susceptibility to SCC of the larynx, pharynx, or oral cavity. The influence of the CCND1 genotype on clinical outcome was also assessed. We found no correlation between genotype and tumor size (T1-T4), the involvement of nodes at presentation, or patient age and gender. However, the distribution of CCND1 genotypes in cases with poorly differentiated tumors was significantly different to that in patients with well-/moderately differentiated tumors (P = 0.016; chi2(2) = 8.71). Homozygosity for CCND1*G (GG genotype) was associated with poorly differentiated tumors (G3). We used Cox's proportional hazards model to investigate the influence of the CCND1 genotype on disease-free interval. CCND1 GG was associated with reduced disease-free interval [P = 0.001; hazard ratio (HR) = 2.95; 95% confidence interval (CI) = 1.54-5.63]. This remained significant after correction for tumor differentiation (P = 0.013; HR = 2.34; 95% CI = 1.2-4.6) and tumor stage (P = 0.005; HR = 2.64; 95% CI = 1.34-5.19). Analysis of the data from patients with tumors at different sites showed that the CCND1 GG genotype was associated with reduced disease-free interval in laryngeal (P = 0.004; HR = 3.63; 95% CI = 1.44-8.83) and pharyngeal (P = 0.006; HR = 3.48; 95% CI = 1.43-8.46) tumors. This is the first report of an association between CCND1 polymorphism and prognosis in SCC of the head and neck. These data show that the CCND1 GG genotype is an independent prognostic indicator of disease-free interval and supports initial observations in non-small cell lung cancer, that polymorphism within CCND1 influences tumor behavior.

Association of arylamine N-acetyltransferases NAT1 and NAT2 genotypes to laryngeal cancer risk.

Genetically polymorphic xenobiotic metabolizing enzymes are supposed to be host factors for an individual's cancer susceptibility. A total of 255 laryngeal cancer patients was genotyped for NAT1 and NAT2 and compared with 510 reference individuals, matched by age and gender. NAT1 genotypes (NAT1*3, *4, *10, and *11 ) were found equally distributed between cases and control individuals. However, there was a significant overrepresentation of 20 (7.8%) homozygous NAT2 genotypes coding for rapid acetylation (NAT2*4/*4 and NAT2*4/*12A) amongst laryngeal cancer patients versus 19 (3.7%) such individuals in the control group (odds ratio 2.18, 95% confidence limits 1.13, 4.22; P = 0.018). Furthermore, an increasing NAT2*4/*4 frequency in cases with strong cigarette consumption was observed, but also in non-smokers. Heterozygous genotypes of NAT2*4/slow were not overrepresented. These results correspond with earlier findings in lung cancer. Analysis of NAT1 and NAT2 combinations revealed a linkage disequilibrium between NAT1*10 and NAT2*4; NAT1*10 frequency was twofold higher in NAT2*4/*4 carriers than in slow NAT2 coding genotypes. In conclusion, the distinct genotype NAT2*4/*4 proved to be a rare, but powerful host risk factor for larynx carcinoma. These data support the notion that an individual's specific NAT2 genotype may be decisive for the organ of his smoking-initiated cancer.

The glutathione S-transferase GSTP1 polymorphism: effects on susceptibility to oral/pharyngeal and laryngeal carcinomas.

We have examined the hypothesis that the polymorphic, glutathione S-transferase GSTP1 gene is a susceptibility candidate for squamous cell cancer of the oral/pharynx and larynx. We describe GSTP1 genotype frequencies in 380 cases and 180 controls. We found a lower frequency of GSTP1 AA in the oral/pharyngeal cases compared with controls (p = 0.003, odds ratio = 0.47) after correction for age and gender. We used an immunohistochemical approach to show widespread expression of the GSTP1 subunit throughout the pharynx and larynx. In uninfiltrated tissue, strong positivity was found throughout the squamous cell epithelium with the exception of the basal cell layer. The cilia of the respiratory epithelium of the larynx also showed positivity for GSTP1. In tumour tissue, expression of GSTP1 was similar in pharyngeal and laryngeal samples. These data are the first to show that polymorphism at GSTP1 mediates susceptibility to squamous cell cancer of the upper aerodigestive tract. No significant interactions were identified between GSTP1 and GSTM1, GSTM3, GSTT1 and the cytochrome P450 CYP1A1, CYP2D6 and CYP1A1 genotypes.

Polymorphism in cytochrome P450 CYP2D6, CYP1A1, CYP2E1 and glutathione S-transferase, GSTM1, GSTM3, GSTT1 and susceptibility to tobacco-related cancers: studies in upper aerodigestive tract cancers.

Glutathione S-transferase GSTM1, GSTM3 and GSTT1 and cytochrome P450 CYP2D6, CYP1A1 and CYP2E1 loci are susceptibility candidates for cancers of the upper aerodigestive tract because putatively protective and risk genotypes have been identified from studies in other diseases associated with alcohol and tobacco consumption. We describe genotype frequencies in 398 oral, pharyngeal and laryngeal squamous cell carcinoma patients and 219 control individuals. Of the genotypes presumed to be protective, only GSTM1 A/B influenced susceptibility; the GSTM1 A/B frequency was lower in the patients than the control individuals both before [odds ratio = 0.3, 95% confidence interval (CI) 0.1-0.7] and after correction for imbalances in age, sex, smoking and alcohol consumption (odds ratio = 0.2, 95% CI 0.1-0.5). Of the putatively risk genotypes, GSTM3 AA, previously associated with susceptibility to skin cancer, was higher in the cases (odds ratio = 1.6, 95% CI 1.1-2.4). Dividing cases into oral/pharyngeal and laryngeal squamous cell carcinoma showed the GSTM3 AA frequency was higher in laryngeal squamous cell carcinoma than control individuals (odds ratio = 1.6, 95% CI 1.1-2.5) and the difference between control individuals and oral/pharyngeal squamous cell carcinoma approached significance (odds ratio = 1.7, 95% CI 1.0-2.8). The putatively protective GSTM3 BB genotype was lower in patients with glottic (1.0%) than supraglottic (3.0%) squamous cell carcinoma. We identified no differences between patients and control individuals in the frequencies of presumed risk genotypes (e.g. CYP2D6 EM, CYP1A1 m1/m1, CYP1A1 Ile/Ile, CYP2E1 DD, CYP2E1 c1c1, GSTT1 null) or, interactions between genotypes and smoking or alcohol consumption. We conclude, first, that mu class glutathione S-transferase influence risk of upper aerodigestive tract cancers thereby complementing studies in skin cancer patients showing GSTM1 A/B is protective, while GSTM3 AA moderately increases risk. The influence of GSTM1 A/B, but not GSTM1 A or GSTM1 B (mostly heterozygotes with GSTM1*0) suggests that two expressed alleles may attenuate risk. While we found immunohistochemical evidence of GSTM3 expression in the cilia lining the larynx, the biochemical consequences of the polymorphism are unclear. Indeed, the influence of the gene may reflect linkage disequilibrium with another gene. However, we did not find an association with GSTM1 genotypes. Second, we conclude that the CYP2D6, CYP2E1, CYP1A1 and GSTT1 alleles studied, although putatively good candidates, either do not determine the effectiveness of detoxification of tobacco-derived carcinogens in the upper aerodigestive tract or, that chronic consumption of tobacco and alcohol overwhelms enzyme defences, irrespective of genotype.

Cyclin D1, glutathione S-transferase, and cytochrome P450 genotypes and outcome in patients with upper aerodigestive tract cancers: assessment of the importance of individual genes using multivariate analysis.

GST, CYP, and CCND1 genotypes have been associated with outcome in several cancers. Accordingly, we have examined, in patients with one squamous cell carcinoma (SCC) of the head and neck, associations between GSTM1, GSTT1, GSTM3, GSTP1, CYP2D6, CYP1A1, CYP2E1, and CCND1 genotypes and the outcome parameters, tumor extension, histological grade, and presence of nodes. We used logistic regression to study, first, each gene individually and, second, in a step-wise model that included all of the genes. Different genes were associated with each outcome parameter. Thus, GSTT1 null was associated with T3/T4 lesions in the oral cavity/pharyngeal (P = 0.029), but not laryngeal, SCC cases. GSTT1 null was also associated with histological differentiation (G3) in the oral cavity/pharyngeal, but not laryngeal, SCC cases, although this association only approached significance (P = 0.069). CCND1 GG was associated with G3 tumors in the oral cavity/pharyngeal (P = 0.011), but not laryngeal, SCC cases. The combination of GSTT1 null/CCND1 GG was also associated with G3 tumors. CYP2D6 PM and HET were associated with lymph node involvement in the laryngeal, but not oral/pharynx, SCC cases. Genes that were individually associated with outcome were also associated with the parameter in the step-wise routine. The GSTT1 null frequency was greater in 39 patients with second primary tumors than in those with one lesion (P = 0.014). The data demonstrate site-dependent associations between GSTT1 null, CCND1 GG, and CYP2D6 PM and tumor extension, differentiation, and nodes.

Local hyperthermia of N2/N3 cervical lymph node metastases: correlationof technical/thermal parameters and response.

PURPOSE: Patients with advanced head and neck carcinomas, primarily nonresectable as well as recurrent cases, were treated in multimodality regimens with radiotherapy, chemotherapy, and local hyperthermia. Commercially available microwave and radiowave applicators were used in 50 patients with N2/N3 cervical lymph node metastases during more than 250 heat treatments. To assess technical suitability, the achieved power densities and thermal parameters were tested for correlation with anatomical and geometrical factors. To assess effectiveness, the response was compared with derived parameters of the achieved temperature distributions. METHODS AND MATERIALS: The temperature measurement points (in thermometry catheters) documented by computerized tomography are labeled according to tissue depth, shielding by osseous structures, and location in relation to the external applicators. Relative and absolute specific absorption rates (SAR) and cooling coefficients are extracted from the temperature-time curves. Time-averaged temperature-position curves are evaluated to obtain index temperatures (T90, T50, T20), minimum/maximum tumor temperatures, cumulative minutes T90 > or = 43 degrees C, and 43 degrees C-equivalent min T90. Radiation dose, treatment time, and chemotherapy regiment are also considered. A response parameter is defined using the pre- and posttherapeutic tumor volumes. A multivariate variance analysis is performed for the dependent variables power density, thermal parameters, and response. RESULTS: A significant correlation exists between power density and absorption, presence of a fat layer, and applicator illumination. The maximum depth is 5 cm, where SAR of >= 10 mW/g are registered. Achieved temperatures at individual measurement points are dependent on the SAR, and to a lesser extent, the perfusion-dependent cooling coefficients, but the index temperature T90 is only significantly related to intratumorally achieved SAR. The thermal gradient (T20-T50) and temperature peak (T20) are significantly influenced by the tumor volume. The response is directly related to the index temperature T90, equivalent minute T90 43 degrees C, and cumulative minutes T90 > or = 40.5 degrees C, and inversely related to the tumor volume. CONCLUSIONS: Local hyperthermia using microwave and radiowave applicators in the head and neck region is a tolerable and clinically practical supplementary therapy used as part of multimodal regimens, and has already been proven to be effective. However, the analyses also demonstrated the limits of currently available technology, and confirm the need for continued methodical research.

Macroencapsulation of human cartilage implants: pilot study with polyelectrolyte complex membrane encapsulation.

Autogenous cartilage transplantation is a generally accepted method in reconstructive surgery. A promising alternative to this established method could be represented by in vitro engineering of cartilage tissue. In both methods of autogenous transplantation, host response induces reduction of transplant size and transplant instability to an unforeseeable extent. To investigate if polyelectrolyte complex (PEC) membranes were able to avoid host-induced effects on implanted tissues without neglecting the tissue metabolism, human septal cartilage was encapsulated with polyelectrolyte complex membranes and subcutaneously implanted on the back of nude mice. Septal cartilage implants, without encapsulation served as control group. Histochemical and electron microscopic investigations were performed 1, 4, 8 and 16 weeks after implantation. In the case of an intact PEC-membrane no interactions between the host and the implant could be observed. In some implants, the capsule was torn in several areas and signs of chronic inflammation with the cartilage having been affected mildly could be observed. Implanted cartilage protected with PEC-encapsulation showed no signs of degeneration and significantly lower level of after effects of chronic inflammation than implanted cartilage without PEC-encapsulation. Therefore, it could be expected, that PEC membrane encapsulation offers a novel approach to protect cartilage implants from host response after autogenous transplantation.

Glutathione S-transferase and cytochrome-P-450 polymorphism as risk factors for squamous cell carcinoma of the larynx.

PURPOSE: While cigarette smoking and alcohol consumption are recognized covariates for laryngeal carcinoma, the role of genetic factors in determining individual susceptibility is unknown. We describe the influence of polymorphism in carcinogen-metabolizing enzymes on susceptibility to squamous cell carcinoma of the larynx. MATERIAL AND METHODS: We investigated 269 patients with T1-T4 laryngeal carcinoma and 216 controls. Enzyme genotypes at the glutathione-S-transferase GSTM1 (A, B, A/B, null), GSTM3 (A, B), GSTT1 (null and expressors), and cytochrome P-450, CYP2D6 (intron 3/exon 4 boundary mutation and exon 5 deletion), CYP1A1 (3'-mutation and exon 7 mutation), and CYP2E1 (mutation at the 5' flanking region) loci were determined using polymerase chain reaction and restriction-based approaches. RESULTS: While the frequencies of the heterozygote GSTM1 A/B and homocygote GSTM3 B/ B were statistically significantly lower in cases than controls, the frequency of the GSTT1 null genotype was higher in the cases than controls. Genotype frequencies of 123 patients suffering squamous cell carcinomas located at different sites within the upper aerodigestive tract showed no differences between cases and controls. CONCLUSIONS: The data provide evidence that susceptibility to laryngeal carcinoma, but not pharyngeal carcinoma, is mediated by allelism at a number of loci encoding enzymes involved in the detoxification of electrophils derived from environmental pollution including cigarette smoke.

[MR-guided laser-induced thermotherapy in tumors of the head and neck region: initial clinical results]. / MR-gesteuerte laserinduzierte Thermotherapie bei Tumoren in der Kopf-Hals-Region: Erste klinische Ergebnisse.

PURPOSE: To analyse the value of MR-guided laser-induced thermotherapy (LITT) for palliative treatment of recurrent tumours of the head and neck region. MATERIAL AND METHODS: 8 patients with recurrent tumours of the head and neck region (squamous cell carcinomas n = 6, pleomorphic adenomas n = 2) underwent MR-controlled LITT. A 7 French laser applicator was inserted under local anaesthesia into the centre of the recurrent tumour. A Nd:YAG laser with a wavelength of 1064 nm was used. Therapy was monitored on-line using special MR thermosequences. RESULTS: Preinterventional contrast-enhanced MRI revealed a recurrent tumour of the head and neck region for all eight patients. All patients tolerated the procedures well under local anaesthesia, with no clinically relevant side effects. The MR thermosequences depicted up to 15 mm diameter areas of less signal near the laser tip. Postinterventional contrast-enhanced MRI revealed hypovascularized areas due to the resulting coagulative necrosis. Coagulative necrosis of 4 cc to 28 cc occurred in all patients, and a reduction of clinical symptoms was achieved in five. CONCLUSION: The results obtained indicate that minimal invasive LITT can be a therapeutic alternative for palliative treatment of head and neck tumours.

Influence of tumour necrosis factor microsatellite polymorphisms on susceptibility to head and neck cancer.

While cigarette smoking and alcohol consumption are recognized covariates for head and neck squamous cell carcinoma (SCC), the role of genetic factors in determining individual susceptibility is unknown. The human tumour necrosis factor (TNF) region on chromosome 6p21 within the major histocompatibility complex (MHC) includes a number of immunologically important genes. Recently, five microsatellite markers have been described in the TNF locus. TNF levels vary with different TNF microsatellite alleles, and associations of these microsatellite markers with autoimmune diseases and different types of cancer have been shown. Therefore, the TNF locus represents candidate susceptibility genes for head and neck cancer. This study describes the influence of TNF a-d microsatellite polymorphisms on susceptibility to head and neck cancer by comparing the allele frequencies of 269 patients suffering from laryngeal cancer and 123 patients suffering from oral cavity pharyngeal cancer and 113 German controls. DNA was extracted from peripheral blood samples, amplified by polymerase chain reaction with fluorescently labelled primers for TNF microsatellite (a-d) and electrophoresed on polyacrylamide gels using an automated DNA sequencer. The data showed no differences in allele frequencies between controls and pharyngeal cancer patients. By contrast, the TNF b3 allele was associated with altered risk for laryngeal cancer (p = 0.0006, odds ratio 2.2). Homozygosity for TNF b3/b3 resulted in an increased risk of developing laryngeal cancer (p = 0.004, odds ratio 5.3). Susceptibility to supraglottic SCC and multiple primary tumours was mediated by the absence of the a11 allele. The data provide the first evidence that allelism at the TNF microsatellite markers alter the risk of developing SCC of the larynx.

Electron microscopy in rhinology.

In rhinology, electron microscopy has been a useful research tool for the past 15 years, but provided only a few direct clinical applications. In this review, the author's work on the human nasal mucosa and the studies of other investigators are discussed, with the emphasis on allergic reactions and disturbances of the autonomous nervous system as well as the immotile cilia syndrome.

Surgery for squamous cell carcinoma of the tongue and floor of the mouth.

Surgery for cancer of the tongue and floor of the mouth has become more varied and generally more conservative, influenced by advances in oncology and modern reconstructive methods. Combined therapy is favored, with postoperative irradiation and sometimes adjunctive chemotherapy, using cis-platinum. T1 carcinomas of the tongue and floor of the mouth can be treated with either wide local excision or irradiation alone, but surgery is the preferred method. T2-T4 tumors treated by resection combined with radiation therapy promise the best results. The indications and principles of the most important operative procedures are discussed: local excision; partial and total glossectomy; excision of the floor of the mouth with marginal mandibular resection; composite resection. Mandible sparing operations such as a modification of the "pull through" technique described by Stell or temporary splitting of the mandible are oncologically safe in many cases. A radical neck dissection is indicated in each carcinoma of the tongue or floor of the mouth with palpable lymph nodes. If no nodes are palpable, an elective neck dissection appears justified in view of the high frequency of clinically occult lymph node metastases. Reconstructive measures following radical tongue and floor of the mouth operations are required for regaining mobility of the remaining tongue, for reconstruction of the floor of the mouth and for replacement of the mandible. For immediate reconstruction, the most frequently used technique is the pectoralis major myocutaneous flap which has largely replaced the previously employed local and regional flaps. A significant problem remains with mandibular reconstruction.

[Technique of microvascular jejunum transfer for replacement of the cervical esophagus]. / Technik des mikrovasculären Jejunumtransfers zum Ersatz des cervicalen Oesophagus.

Technique and results of the first 10 free jejunal grafts after pharyngolaryngectomy are presented. End-to-side venous anastomosis and end-to-side pharyngojejunostomy instead of end-to-end techniques are proposed. Hospital mortality was 0 and as minor complications 1 uncomplicated fistula of the proximal and 1 stenosis of the distal anastomosis along with 1 wound infection were seen. There were no vascular complications and no graft loss. All patients were able to swallow and to eat thus an excellent quality of life was achieved.

[Diagnostic approach to dysphagia from the ENT viewpoint]. / Diagnostisches Vorgehen bei Dysphagie aus HNO-ärztlicher Sicht.

The diagnostic procedure for the symptom oropharyngeal dysphagia is described from an otorhinolaryngologic point of view, with emphasis on an interdisciplinary team approach. The various methods used in modern otorhinolaryngology both by the practitioner and the clinician are discussed with regard to a step-like diagnostic approach. The diagnosis should be made on the basis of aetiology, anatomy and function. First all organic changes causing oropharyngeal dysphagia must be recognized, both "typical ENT diseases" and malignant tumors of the oropharynx and hypopharynx which are usually diagnosed only in an advanced stage. If there is no evidence of an organic cause in the field of otorhinolaryngology, as a differential diagnosis a dysfunction of the upper esophageal sphincter and a "globus pharyngus" have to be ruled out.