[Dilatational Tracheotomy in Head and Neck Surgery]. / Der Einsatz der perkutanen Dilatationstracheotomie im Rahmen der Kopf-Hals-Tumorchirurgie.

OBJECTIVE: The pre- and postoperative airway management during surgery of head and neck cancer is a clinically relevant and challenging task. Usually an epithelialised tracheostomy is used. The significance of percutaneous dilatational tracheostomy (PDT) in combination with tumour surgery of the head neck area has not yet been fully considered. MATERIAL AND METHODS: Within a surgical therapy of head neck tumours, 58 patients were treated with a PDT by Ciaglia or Fantoni at the Department of Otorhinolaryngology, Head and Neck Surgery, Martin-Luther-University Halle during the period from December 2002 to October 2007. The patient data were retrospective analyzed in a pseudonymous way. In a follow-up examination the resulting conditions of the PDT were reviewed. RESULTS: The median of the length of time till decannulation was 12 days. There were no documented disorders or complications after PDT at 25 patients (43%). As serious complications pneumothorax in 2 patients (3,4%), 9 bleedings (15,5%) and 1 infection (1,7%) were recorded. After occurrence of tracheostomy metastases in 2 patients with tracheostomy by Fantoni, this method was no longer used. For the subsequent follow-up examination 33 patients were included. No tracheal stenosis or serious long term complications were determined. CONCLUSIONS: Under consideration of the indications and contraindications, the percutaneous dilatational tracheostomy by Ciaglia with associated bronchoscopy is a safe method with low complication rate for the airway management of patients with head neck cancer. Long term complications and disorders after PDT are not to be expected. Tracheotomy according to Fantoni cannot be recommended for this treatment.

[Atypical carcinoid of the larynx. Case report and review of literature]. / Atypisches Karzinoid des Larynx. Fallbericht und Literaturübersicht.

BACKGROUND: With an incidence of over 90%, squamous carcinomas are the most frequent laryngeal malignancies. Laryngeal neuroendocrine tumors appear only in approximately 1%. These are aggressive tumours with a high rate of metastases. To date, no diagnosis or treatment guidelines have been established for this entity. CASE REPORT: We describe the case of a 63-year-old man complaining of dysphagia and intermittent hoarseness. Initial laryngoscopy showed a tumour at the laryngeal epiglottic surface. Staging demonstrated an atypical carcinoid tumour with local lymph-node metastasis. At 24 months following revision surgery with safety distance, elective bilateral neck dissection and adjuvant radiotherapy the patient is free of complaints and recurrence. CONCLUSION: In atypical carcinoid tumours total resection as well as bilateral neck dissection should be performed, even in cN0 cases. In the presence of cervical lymph node metastases adjuvant radiotherapy is indicated.

[Therapeutic management of dislocated laryngeal fractures]. / Therapeutisches Vorgehen bei dislozierten Larynxfrakturen.

BACKGROUND: Acute trauma with laryngeal fracture is a rare injury affecting all functions of the larynx. Resuscitation follows the ABC principles for acute trauma life support. The priority has always been the establishment of an adequate and secure airway. To achieve optimal therapeutic long-term results, permanent stabilisation of the airway and good functional restoration of phonation and swallowing are necessary. We describe the therapy and long-term follow-up of two patients who suffered laryngeal fracture. CASE REPORTS: The first case concerns a 31-year-old woman who suffered polytrauma with laryngotracheal separation in a car accident. The fracture was reduced and fixed utilizing miniplates. After decannulation the patient had a patent airway. She regained normal voice and was able to swallow without difficulty. The other case concerns a 16-year-old boy who suffered a laryngeal fracture in a bicycle accident. He also underwent subsequent operative therapy using adaptation plates. This patient also regained an adequate airway and a good postoperative voice. There were no problems with swallowing. CONCLUSION: Chondrosynthesis with adaptation plates provides an enduring and exact fixation of laryngeal fractures. In the long term, this leads to a stable recovery of function concerning airway, voice, and swallowing.

[Large endolymphatic duct and sac syndrome : part 2: clinical manifestations]. / Der erweiterte Ductus und Saccus endolymphaticus : Teil 2: Klinische Manifestationen.

OBJECTIVE: The aim of this study was to clarify the clinical course of large endolymphatic duct and sac syndrome (LEDS). Although LEDS is the most common form of radiologically detectable inner ear malformation associated with sensorineural hearing loss (SNHL), it is relatively unknown in the German-speaking countries. We hoped to derive useful pointers for clinical care. METHODS: We evaluated the clinical audiometric records and imaging findings of 169 patients with clinically suspected inner ear malformation seen between 1994 and 2003. Following identification of all LEDS cases, we serially investigated the anamnestic and functional data with regard to severity and course. RESULTS: Among 169 patients, 17 (median age 12 years; 12 females) showed LEDS. A total of 28 ears were affected. Clinically, most cases were of prelingual or perilingual onset and displayed steady or fluctuatingly progressive severe SNHL with emphasis on the high frequencies or deafness. Episodes of sudden hearing loss were relatively frequent. No correlation could be demonstrated between the severity of morphological changes and the degree of SNHL. CONCLUSION: Progression of SNHL and episodes of sudden hearing loss were seen mainly in late childhood or adolescence. This should be taken into account when informing and advising patients and planning therapy.

[Large endolymphatic duct and sac syndrome (LEDS) : part I: analysis of imaging findings]. / Der erweiterte Ductus und Saccus endolymphaticus : Teil 1: Analyse bildgebender Befunde.

OBJECTIVE: Large endolymphatic duct and sac (LEDS) syndrome is the most common radiologically detectable form of inner ear malformation (IEM) associated with sensorineural hearing loss (SNHL). Nevertheless, it is relatively unknown in the German-speaking countries. METHODS: Among all patients referred from ear, nose, and throat specialists to the radiology department with clinically suspected IEM between 1994 and 2003, we identified the cases of LEDS. The retrospective study included clinical records, high-resolution computed tomography, and magnetic resonance imaging. RESULTS: Of 169 patients, 17 (median age 12 years; 12 females) showed LEDS. A total of 28 ears were affected. Ten patients (6%; 15 ears) had isolated LEDS, while seven patients showed additional IEM (4%; 13 ears). The most frequent of these was dysplastic vestibule (13/13 ears), followed by Mondini deformity (10/13 ears) and dilated semicircular canals (7/13 ears). Three of 13 ears revealed severe IEM of the cochlea, vestibule, and semicircular canals. No correlation could be demonstrated between the severity of morphological changes and the degree of SNHL. CONCLUSION: LEDS may be an underestimated cause of SNHL. Imaging is necessary for a confident diagnosis.

Expression and distribution of cytokeratins and vimentin in rat larynx and trachea following irradiation.

BACKGROUND: External irradiation (IRR) of advanced head and neck tumors often includes tissues of the larynx and trachea unaffected by cancer. In these normal tissues, both single-cell damage (necrosis, apoptosis, functional cell death) and interstitial damage (edema, fibrosis, vascular alterations, cellular infiltrations) resulting in tissue remodeling can occur, depending on various IRR parameters. However, reports on radiogenic intermediate filament protein alterations in laryngeal-tracheal tissues are very rare. In this study, we investigated the phenotypic characterization of the normal integrity-supporting cytokeratins (CK) and vimentin following a clinically relevant IRR protocol in laryngeal-tracheal tissues. MATERIALS AND METHODS: In 61 laryngo-tracheal specimens from Wistar rats the expression profile and distribution pattern of CK (CK13, CK17/19, CK18) and vimentin were investigated according to IRR dose (fractionated IRR, 2 Gy per day, total dose of 20, 40 or 60 Gy), time from IRR (6 months vs. 12 months) and animal age (1 year vs. 1.5 years) using immunohistochemical methods, semiquantitative assessment and multivariate analysis. RESULTS: In irradiated specimens, expression of both CK and vimentin showed slight to moderate dose-dependent alterations. The expression differed in frequency and level among the various tissue structures and showed remarkable heterogeneity, with increases, decreases and fluctuations in staining. In the glottic mucosal layer (non-keratinizing squamous epithelium), CK13 expression decreased with increasing dose. The CK17/19 expression of supra- and subglottic respiratory epithelia following 20 and 60 Gy exposure was significantly lower than in controls. The respiratory epithelia and, in part, the cuboidal epithelia of the indifferent type at the inner side of the aryepiglottic fold revealed increasing CK17/19 immunoreactions up to 40 Gy IRR, but a distinct decrease in expression at 60 Gy. In subglottic gland structures, CK18 was detected at significantly higher levels than in controls. There was increasing expression with increasing dose. CK18 reactions of supra- and subglottic respiratory mucosal layer, supraglottic gland structures and thyrocytes tended towards increasing expression with increasing dose and in older animals. Tracheal mucosal epithelia, tracheal glands, and respiratory epithelia of the inner side of the aryepiglottic fold tended towards decreasing expression of CK18 with increasing dose and in older animals. In part, these tissues showed dose-dependent fluctuations. Furthermore, the vimentin reactions showed dose-dependent, heterogeneous patterns, with increases, decreases, and fluctuations in staining. Moreover, there were differences in frequency and intensity of expression among the various tissue structures. Age and time from IRR had no significant effect on immunoreaction. CONCLUSION: The staining of CK and vimentin predominantly showed a notable dose-dependent heterogeneity, with increases, decreases and fluctuations in expression. The expression pattern persisted for up to 1 year after the completion of irradiation. Thus, these findings must reflect late radiation effects. The altered expression of CK and vimentin may play at least a partial role in structural (e.g. edema) and functional (e.g. voice disorders) changes associated with irradiation of the head and neck.

Rat tumors following fractionated irradiation.

BACKGROUND: Rats are susceptible to irradiation and can develop benign and malignant tumors either spontaneously or in the field of irradiation. In the head and neck region, there are no reports available on the type of tumor after fractionated irradiation using a human therapy protocol. MATERIAL AND METHODS: We analyzed 19 tumors, in 19 rats, which developed after external X-irradiation of the left neck area in Wistar rats (2 Gy/day, monofractions, 5 days/week, total dosage 60 Gy) and compared the findings with tumors in untreated rats of the same strain. RESULTS: Tumors in the irradiation field proved to be squamous cell carcinoma or adenoid cystic carcinoma (ACC), not sarcoma. These entities were sporadically found in non-irradiated rats at a higher age. CONCLUSIONS: ACC has rarely been reported in the literature on laboratory rats. The development of this highly aggressive malignant tumor can be expected 3 months to 1 year after completion of irradiation.

Expression of cytokeratins and additional markers in undifferentiated lymph node metastases of the neck.

OBJECTIVE: A variety of carcinomas (CA) can metastasize to the lymph nodes of the neck. Differentiation of CA in the lymph nodes according to their resemblance to the structure of origin is the basis of histopathological diagnosis. However, during the course of the disease, e.g. tumor recurrence after ablative surgery, these tissues can completely lose the ability to imitate typical structures of the organ that gave rise to malignant transformation. This can result in the inability of the pathologist to identify the origin of the metastases. The identification of the large group of cytokeratins (CK) as a member of the family of intermediate filaments has improved the diagnosis of epithelial tissues. An attempt was made to use CK antibodies to identify the organ of origin of poorly differentiated and anaplastic lymph node metastases of the neck. METHODS: We investigated 34 routinely fixed (formalin/paraffin) lymph node metastases of the neck or specimens of these metastases. The tumors differed in terms of suspected primary tumor site and differentiation. Depending on the case history, diagnosis was performed by hematoxylin-eosin staining and by immunohistochemical staining of sections using antibodies against CK (CK nos. 1-10-11, 5-6, 6-8, 7, 8, 8-18, 13-15-16, 19 and 20) and against additional markers [vimentin, leukocyte common antigen (LCA), S-100, gross cystic disease fluid protein (GCDFP) and Epstein-Barr virus-induced latent membrane antigen (EBV-LMP)]. RESULTS: The histopathological diagnosis was lympho-epithelial CA (8 cases), thyroid gland CA (2 cases), mammary gland CA (5 cases), bronchial CA (4 cases), basaloid CA (3 cases), clear cell CA (2 cases), sebaceous CA (1 case) and pharyngeal CA (9 cases). Some metastases were anaplastic in differentiation (G3-4). The marker expression in the immunohistological sections supported the histopathological findings. In some cases diagnosis succeeded especially in evaluating the marker expression. For example, in lympho-epithelial CA the epithelial tumor cell formations were positive for CK 5-6 antibodies, while expression in the lymphatic cells was lacking. In mammary gland CA the tumor cells were clearly identified by their positivity for CK 7 and GCDFP antibodies, in contrast to the negative infiltrating cells surrounding the tumor cells. CONCLUSIONS: In light of the case history details and the histopathological findings, immunohistological expression of various markers, especially the identification of CK subtypes, supported the differential diagnosis. In some cases the diagnosis may be established only by using CK profile and additional markers.

Distribution of collagens in carcinomas of salivary and mammary gland origin in irradiated rats.

OBJECTIVE: While radiation-induced sarcomas or carcinomas following chemical carcinogens are well-documented in rats, radiation-induced carcinomas, especially adenoid-cystic carcinomas (ACC) and adenocarcinomas, originating from the head and neck region, including the major salivary glands (SG) are rarely reported. Because in human ACC of the SG structural changes of the basement membrane (BM) with positive correlation of tumor differentiation loss and BM thinning have been described, this study set out to analyze collagen distribution in malignant rat tumors (TM) developing inside the radiation field (RF) and spontaneously. The TM arose in the course of studies on other questions regarding radiation effects following fractionated radiation (2 Gy/day, 5 times a week; total dose 60 Gy). METHODS: We investigated 22 TM (14 malignant, 8 benign) of 22 female Wistar rats. The RF comprised the left head and neck area. Besides assessment of hematoxylin-eosin (HE)-stained sections collagens (C; types III and IV) were investigated using immunohistochemical methods. RESULTS: Nine malignant TM originated from the SG, a further three from the milk line and two from the maxilla. Two ACC, two cystadenocarcinomas, one microcystic adenocarcinoma and four squamous cell carcinomas (SCC) arising from the SG (one SCC was observed in the maxilla) developed in the RF. One microcystic adenocarcinoma, one ACC and one adenocarcinoma with sebaceous differentiation arising from the milk line and one SCC arising from the maxilla were found in non-irradiated animals. As typical results, in the ACC (glandular subtype), C III was detected in the interstitium with sometimes stronger staining surrounding myoepithelial cells (MC) and excretory duct structures (ECD). Weak C IV staining in a string-like fashion was found in ECD and MC. In larger pseudocysts the lumen contained substances reacting with C IV antibodies. In the cystadenocarcinomas and the microcystic adenocarcinomas reactions at variable levels after anti-C III incubation were found close to modified MC and ECD with transition to the interstitium. C IV was more intensely stained in these entities, in part continuously and with broadening around MC and ECD. However, especially in more anaplastic parts of the tumor, fragments, interruptions or loss of the BM were noted. Focal interstitial immunoreactivity, e.g. conglomerates, was also identified. CONCLUSION: In rat carcinomas collagen detection was partially of a continuous layer, even with BM thickening and more extended deposition. In contrast, BM fragmentation or loss was displayed more often in anaplastic parts of the tumor. Also, the interstitium showed conglomerated collagen formations. Therefore, the increasing loss of BM in rat SG tumors is similar to that in humans and in both species is a sign of dedifferentiation.

Cytokeratin expression in carcinomas of irradiated rats.

OBJECTIVE: Laboratory rats can develop benign or malignant tumors (TM) spontaneously or following various carcinogenic processes, e.g. irradiation. The effects of irradiation vary according to the irradiation field (RF), the dosage and the strain of rat. Radiation-induced malignant TM in rats are predominantly sarcomas. Carcinomas, especially adenoid-cystic carcinomas (ACC) and adenocarcinomas of the head and neck region, are rarely reported in rats. The aim of this study was to add to the knowledge on ACC and adenocarcinomas in rats developing inside the RF and spontaneously. The TM arose in the course of studies on other questions of radiation effects following fractionated irradiation (2 Gy/day, 5 times a week up to a total dose of 60 Gy). METHODS: We investigated 22 TM (14 malignant, 8 benign) of 22 female Wistar rats. Ten malignant TM developed in the RF and 4 outside of the left head and neck area. The RF comprised the left neck, extending from left auricle to left clavicle and included the midline organs of the neck. Besides assessment of hematoxylin-eosin (HE)-stained sections, epithelial differentiation was investigated using cytokeratin (CK) antibodies against CK 5/6, CK 7, CK 8/18, CK 13/15/16, CK 17 and CK 20 and the LSAB-2 detection system. RESULTS: Nine malignant TM originated from the major salivary glands (SG), a further three from the milk line and two from the maxilla. Using HE staining the pattern of rat malignant TM differed from that found in humans and was difficult to interpret. Two ACC, two cystadenocarcinomas, one microcystic adenocarcinoma and four squamous cell carcinomas (SCC) arising from the SG (one SCC was observed in the maxilla) developed in the RF. One microcystic adenocarcinoma, one ACC and one adenocarcinoma with sebaceous differentiation arising from the milk line and one SCC arising from the maxilla were found in non-irradiated animals. As typical results, in the ACC CK 17 was distinctly immunoreactive in excretory duct structures (ECD). CK 5/6 and CK 13/15/16 were marked at variable levels in myoepithelial cells (MC) and in basal cells of ECD. In the cystadenocarcinomas the ECD were clearly identified with CK 17 and CK 8/18 antibodies. MC and basal cells of ECD were positive for CK 5/6 and CK 13/15/16 antibodies. CONCLUSION: The CK expression profile of these rare and aggressive TM in rats differed according to the entity and SG structure. The differentiation markers were predominantly found in ECD and in modified MC. Concerning the growth pattern of the TM, the variation in size of the cysts and pseudocysts was remarkable. The unusual tumor features reduced the comparability with humans. The differentiation pattern did not differ noticeably between TM originating inside or outside the RF. Identification of CK subtypes in rat tumors facilitates their differential diagnosis.

Dose-response relationships and the effect of age and latency period on the expression profile of laminin in irradiated rat mandibular glands.

OBJECTIVE: The extent of radiogenic salivary tissue damage (direct cell damage, reactive cell infiltration, increasing fibrosis) depends on the radiation dose (RD), the fractionation (FN) and the localisation of the salivary gland (SG) in the radiation field (RF). Because the histological damage is characterized by cellular and structural alterations and because extracellular matrix proteins, for example laminin (LA), are among the main components of the basement membrane (BM), which is considered to possess cell- and tissue structure-regulating functions, we set out to describe radiogenic BM changes. MATERIALS AND METHODS: In 124 rat mandibular SG we investigated HE-stained sections and the LA staining profile (LASP) dependent on age [1 year (y) vs. 1 1/2 y], on FN [2 Gy/day up to a total dosage of 20/40/60 Gy (X-rays)], on RF [(left head and neck area); inside (left-sided SG) vs. outside (right-sided SG) RF] and on the latency of irradiation (1/2 y vs. 1 y) using immunohistochemical methods and multivariate analysis. RESULTS: The immunoreactivity of the polyclonal antibody in the rat did not differ from the LASP in humans. LA was detected at variable levels in the glandular tissue, in vascular walls (without staining in the adventitia) and in nerve tissue (NT). No reaction was found in the capsule and the trabecula. In irradiated SG most tissue structures, including the vascular wall and NT, showed significantly stronger staining (both increased intensity and more extended expression). With increasing RD from 20 Gy to 60 Gy we found an increasing staining reaction. In addition, conglomerated LA depots were noted, especially in ares of disturbed or former acini of glands exposed to 60 Gy. The LASP up to 20 Gy was non-uniform and did not differ significantly from that in controls. A remarkable finding was the similar dose-dependent increase in staining reaction on both sides in several tissues of irradiated animals. Predominantly, however, we found considerably greater increases in staining in directly exposed glands than in out-of-field glands. Age and time since irradiation had no significant effect on staining. CONCLUSION: The BM constituent LA showed dose-dependent increases in expression--analogous to positively dose-dependent increasing SG tissue damage--which could contribute to disturbed cell-matrix interaction and SG radiation damage. Several parenchymal structures may be more sensitive to possible scattered radiation.

Immunohistochemical detection of cytokeratins in the irradiated rat mandibular gland.

UNLABELLED: BACKGROUND/MATERIAL AND METHODS: The present study is a prerequisite for the exploration of the cytokeratin (CK) profile in rat salivary gland tumors. In 59 rat mandibular glands we investigated the distribution of CK by immunohistochemical analysis. The animals differed in age and pretreatment status (irradiation versus no irradiation). RESULTS: The monoclonal antibodies (CKE3, Ks13.1, NCL5D3, K8.12; against CK 17, CK 13, CK 8 and CK 13/15/16, respectively) identified different epithelial structures in rat salivary gland tissue, including intercalated duct cells (ICD), striated duct cells (SD), granular convoluted tubules (GTC), excretory duct cells (ECD) and myoepithelial cells (MC). As typical results, CKE3 usually stained the ICD, SD and ECD moderately to strongly and stained the GTC slightly. K8.12 staining was restricted to ECD and MC. Differences in immunoreactivity were seen between irradiated and non-irradiated glands, predominantly with stronger staining in the irradiated group. CONCLUSIONS: Previous radiation has to be considered when interpreting immunohistochemical stainings of salivary gland tissue, especially in tumor differentiation studies following irradiation.

Immunohistochemical detection of extracellular matrix proteins in the irradiated rat mandibular gland.

UNLABELLED: BACKGROUND/MATERIAL AND METHODS: In 59 rat mandibular glands we investigated the distribution of extracellular matrix proteins (EMP) in rat salivary gland tumors. The animals differed in age and pretreatment status (irradiation versus no irradiation). For immunohistochemical analysis we used polyclonal EMP antibodies [anti-laminin (AL), anti-fibronectin (AF), anti-collagen-III (AC-III), anti-collagen-IV (AC-IV)]. RESULTS: The antibodies identified the different structures of the extracellular matrix: components of basement membranes [(BM); laminin, collagen IV] and components of BM-associated structures (fibronectin, collagen III). Immunoreactivity was found in glandular tissue, in vascular walls, in nerve tissue and the interstitial connective tissue. Various EMP showed different patterns. Significant differences were seen between irradiated and non-irradiated glands, often with generally stronger and more extended staining in the irradiated group. Thus, both nerve tissue and excretory ducts (ECD) were in part strongly stained for AL, while ECD were intensely positive for AF. CONCLUSIONS: Preceding radiation has to be considered when interpreting salivary gland tissue, especially in tumor differentiation studies following irradiation.

The expression pattern of collagen I in irradiated mandibular salivary glands of rats.

UNLABELLED: Irradiation damage to salivary glands leads to loss of function and fibrosis. Immunohistochemical analysis of extracellular matrix proteins might give a more precise insight into the irradiation damage of glands. Collagen I (C-I) is a major component of the extracellular space. The aim of this study was to analyse the effects of irradiation on the distribution pattern of C-I in the salivary glands of rats. MATERIALS AND METHODS: Sixty female Wistar rats were fractionated irradiated up to 60 Gy (left side of the neck; 2 Gy/d, 5d/week; total dosage either 20, 40 or 60 Gy). The glands were explanted after 6 or 12 months following supravital anesthesia, the shielded right gland serving as internal control. C-I was detected immunohistochemically. RESULTS: In non-irradiated animals the immunoreaction was mainly homogeneous and slight around the ductal epithelia, in the area of the capsule and septae and the peri- and epineurium of nerves. A statistically significant difference was identified in the irradiated rats vs control animals and comparing in-the-radiation field (left side) vs outside-the-radiation field (right side) situated glands. Multivariate analysis revealed a statistically significant increase in staining of irradiated rats concerning the excretory ducts, the area of the capsule and septae, the nerves and striated ducts and adventitia of vessels [p = 0.0001]. The increase of immunoreaction in irradiated glands started above 20 Gy total dosage and was at its maximum after 60 Gy. However, the expression profile was inhomogenous following 20 Gy exposure and did not differ statistically from glands of control animals. Neither the age of the animals nor the latency period following exposure to the radiation source yielded a statistically significant effect on the immunoreaction. CONCLUSION: Studies on irradiation damage to the salivary glands require a more detailed description of the proteins accumulating in the extracellular space, thereby forming the so-called "fibrosis". These accumulations of proteins, e.g. C-I, may both support apoptosis and support a hypoxic environment giving rise to transformed cells.

Dose-response relationships on the expression profile of cytokeratins and vimentin in rat submandibular glands following fractionated irradiation.

OBJECTIVE: The extent of radiogenic damage in salivary gland (SG) tissue depends on the radiation dose (RD), the fractionation (FN) and the localization of SG in the radiation field (RF). While the functional restriction and the radiogenic SG tissue damage are well documented using histomorphological, electron-microscopic and enzyme-histochemical methods, immunohistochemical analysis (IH) of cytokeratins (CK), epithelial differentiation markers, and vimentin, a marker of mesenchymal cells, are rare. Previous studies have shown stronger immunoreactivities of CK in irradiated glands exposed to 60 Gy total dosage. This study was performed to examine dose dependence and alterations related to age, RF, and latency of irradiation. METHODS: In 124 rat mandibular SG we investigated the vimentin and CK staining profile dependent on age [1 year (y) vs. 1 1/2 y], on FN [2 Gy/day up to a total dosage of 20/40/60 Gy (x-rays)], on RF (inside vs. outside RF) and on the time since irradiation (1/2 y vs. 1 y) using IH. RESULTS: The mouse monoclonal anti-CK antibodies [(AB) D5/16B4, Ks 13.1, E 3, K8.12, Ks 18.04, against CK 5-6, CK 13, CK 17, CK 13-15-16, CK 18) and the polyclonal anti-vimentin AB GP53 identified different epithelia and mesenchymal structures in rat SG tissue, including excretory duct cells (ECD), striated duct cells (SD), granular convoluted tubules (GTC), intercalated duct cells (ICD) and myoepithelial cells (MC). MC and mesenchymal cells were positive for vimentin AB. The different CK were detected in cell type-specific patterns and at variable levels in non-irradiated SG. In irradiated SG most cell types showed significantly stronger staining for various CKs. With increasing RD from 20 Gy to 60 Gy we found an increasing staining reaction. The CK staining profile up to 20 Gy was non-uniform and did not differ significantly from controls. Age and time since irradiation played a minor role or had no significant effect on staining. CONCLUSIONS: The CK and vimentin immunoreactivity showed dose-dependent increasing expressions, which could contribute to radiogenic cell and tissue damage. In some tissue structures a possible scattered irradiation effect should be mentioned. Age and time since irradiation (chosen in the study) had a minor or insignificant effect on staining profiles.

Epstein-Barr virus DNA, intermediate filaments and epithelial membrane antigen in nasopharyngeal carcinoma.

The aim of this study was to investigate the expression of intermediate filaments (cytokeratin, vimentin), epithelial membrane antigen (EMA) and the presence of Epstein-Barr virus (EBV) DNA in undifferentiated nasopharyngeal carcinoma (NPC). A high incidence of nuclear signals in NPC was found in primaries and regional lymph node metastases (70%), using 35S-labelled probes of EBV plasmids for in-situ hybridization. Keratinizing squamous cell carcinomas were EBV-negative. All carcinomas were immuno-reactive for cytokeratin (KL-1). 45% of the carcinomas were positive for vimentin. The expression of epithelial membrane antigen was restricted to epithelial cells and reduced in NPC as compared to the distribution pattern of cytokeratin. Both EBV DNA and vimentin in NPC were present in 9 cases. However, in 5 cases NPC were harboring EBV but were not immunoreactive for anti-vimentin antibodies. In no case was a vimentin-positive NPC also EBV-negative. The identification of cytokeratin subtypes revealed no specific cytokeratin pattern in NPC. The expression of vimentin in NPC is not specific for EBV, but seems to reflect the loss of inter-epithelial contact in anaplastic carcinomas.

P53 auto-antibodies in the sera of patients with oral squamous cell carcinoma.

AIM: To investigate the sera of patients with oral squamous cell carcinoma (OSCC) for the existence of p53 auto-antibodies. MATERIAL AND METHODS: The sera of thirty-nine OSCC-patients were investigated. All samples were from untreated patients with no history of another neoplastic disease. The sera of nine healthy subjects served as controls. RESULTS: P53 auto-antibodies were not detected both in all healthy subjects and in 20.6% OSCC-patients with their disease at an advanced stage. We prove that the extinction rates for p53 auto-antibodies in OSCC are very weak in the majority of investigated sera (50%) and are not related to tumour stage. CONCLUSION: It is unlikely that p53 auto-antibodies can serve as a prognostic marker of OSCC.

TNM staging, histopathological grading, and tumor-associated antigens in patients with a history of mucoepidermoid carcinoma of the salivary glands.

BACKGROUND: The aim of this study was to evaluate the TNM categories and histopathological grading as prognostic factors in mucoepidermoid carcinoma (MEC). In addition, the study was designed to provide baseline data on levels of tumor-associated antigens (TAA) in sera of MEC patients. MATERIALS AND METHODS: Fifty-nine patients with MEC of the salivary glands were evaluated. RESULTS: Reclassified TNM stage, at the time of initial diagnosis, varied considerably. In disease-free patients, none of the tested sera were TAA elevated above the cut-off levels. Our patients who died of tumor metastasis had all been classified as stage III or IV at the time of initial diagnosis. Distant metastases are rarely found even decades after surgical therapy. CONCLUSIONS: Long-term follow-up is recommended for patients with MEC of the salivary glands. TAA are not elevated in disease-free patients. The value of TAA in the monitoring of MEC patients remains to be evaluated in further studies.

Expression and distribution of basement membrane proteins in rat larynx and trachea following irradiation.

OBJECTIVE: Basement membranes-(BM) influence polarization, differentiation, migration and proliferation of cell and play an important role in maintaining structural and functional tissue integrity. While BM alterations have been reported in various lesions (e.g. inflammation, tumors) of laryngeal-tracheal tissues, reports on radiogenic BM alterations are rare. External irradiation (IRR) of advanced head and neck tumors often includes "normal tissues" (tissues without cancer) of the larynx. In these normal tissues both single-cell damage (necrosis, apoptosis, functional cell death) and interstitial damage (edema, fibrosis, vascular alterations, cellular infiltrations) resulting in tissue remodeling can occur, depending on various IRR parameters. In this study, we set out to add to our knowledge on the phenotypic characterization of the radiogenic BM expression pattern in laryngo-tracheal tissues. MATERIALS AND METHODS: In 63 laryngo-tracheal specimens from Wistar rats, we investigated the laminin (LA) and collagen IV (CIV) expression profile and distribution pattern depending on the IRR dose (fractionated IRR, 2 Gy/day, up to a total dose of 20, 40, or 60 Gy), the time since IRR (6 months vs 12 months) and animal age (1 year vs 1.5 years) using immunohistochemical methods, semiquantitative assessment, and multivariate analysis. RESULTS: In specimens irradiated with more than 20 Gy, both BM constituents predominantly showed dose-dependent increases and sometimes fluctuations in staining at slight to moderate levels. The expression differed in frequency and level among the various tissue structures. In some structures there was decreased expression. In the vocalis muscle, laryngeal and esophageal nerve endings, recurrent laryngeal nerve and laryngeal and tracheal muscles, LA was detected at levels significantly stronger than in controls. BM surrounding gland structures, nerve endings of the piriform sinus and esophageal muscles showed a marked tendency towards increased LA expression. However, the BM underlying the mucosal layer of the supra- and subglottic region revealed decreasing LA immunoreaction up to 40 Gy IRR, but a distinct increase in expression at 60 Gy. In the esophageal and tracheal muscles, tracheal perichondrium, recurrent laryngeal nerve and capillaries, CIV was detected at significantly stronger levels than in the controls. The vocal ligament exhibited positive CIV immunoreactions adjacent to interstitial and infiltrate cells and CIV-positive BM condensations, resulting in increased staining of these structures. CIV reactions of laryngeal and hypopharyngeal nerve endings tended towards increased expression. In contrast, BM staining surrounding vocal muscle cells revealed significantly decreased expression. In addition, there was a tendency towards decreased expression for supraglottic, subglottic and hypopharyngeal muscle cells. Age and time since irradiation had no significant effect on staining. CONCLUSION: The BM constituents laminin and collagen IV showed prominent dose-dependent increases and sometimes fluctuations in expression. This expression pattern persisted up to one year after completion of the irradiation. Thus, these findings must be related to late radiation effects. The altered BM expression may play a role, at least in part, in structural (e.g. laryngeal edema) and functional (voice disorders) changes associated with irradiation of the head and neck area.