Determination of HER-2/neu amplification and expression in tumor tissue and cultured cells using a simple, phenol free method for nucleic acid isolation.

A rapid, simple and non-toxic procedure for the simultaneous isolation of DNA and RNA from tumor tissue and cells grown in vitro is described. Guanidinium isothiocyanate was used for homogenization of tumor tissue and for cell lysis. Separation of proteins, DNA and RNA was carried out by isopycnic centrifugation in cesium trifluoroacetate. DNA was further purified by salting out residual protein. Nucleic acids prepared by this method from 47 primary human carcinomas and 17 human cell lines were analysed for amplification and expression of the HER-2/neu proto-oncogene. 2- to 10-fold amplification of HER-2/neu was noted in 7/22 mammary carcinomas (32%) and in 4/14 ovarian carcinomas (28%). No amplification of the proto-oncogene was found in 4 laryngeal carcinomas, 1 pharyngeal carcinoma, 2 retrolingual carcinomas, 3 gastric carcinomas and 1 kidney carcinoma. HER-2/neu overexpression was observed in 6/22 of mammary carcinomas (27%) and 7/14 of ovarian carcinomas (50%). No overexpression was found in all other carcinomas studied. Concordance between amplification and overexpression was noted in 3 mammary and 4 ovarian carcinomas, respectively. 3 mammary and 3 ovarian carcinomas showed overexpression without amplification. 5 human mammary carcinoma cell lines showed both amplification and overexpression of HER-2/neu. In two mammary carcinoma cell lines (MDA MB-453 and ZR 75-1) overexpression was noted without amplification of the proto-oncogene. These data combine to suggest that mechanisms other than gene amplification may also lead to overexpression of the HER-2/neu protooncogene in cancer cells.

Incidence of cancer of the respiratory and upper digestive tract in urban and rural eastern Austria.

The incidence of head and neck, oesophagus and lung cancer between 1981 and 1985 was studied in Eastern Austria for an urban-rural division. In males, rural rates of oral cavity, oropharynx and oesophagus tumours were higher than urban rates. For lung tumours, urban rates slightly exceeded rural rates. In females, the incidence of oral cavity, oropharynx, larynx, hypopharynx, oesophagus and lung cancer showed an urban predominance, steepest for head and neck and oesophagus cancers. Cancer of the oral cavity, pharynx, larynx, oesophagus and lung had a high male preponderance.

Expression of cathepsin D in head and neck cancer.

To determine overexpression of cathepsin D in head and neck tumours we examined cytosols from 53 primary tumours, nine cytosols of lymph node metastases and 12 cytosols from adjacent normal tissue. We found a significantly lower concentration in normal tissue compared with tumour cytosol as well as with metastases, even when we compared tumours and corresponding metastases pairwise. In addition, we found a significantly higher concentration of cathepsin D in five lymph node metastases than in the corresponding tumours. We conclude that the reported role of cathepsin D is not restricted to breast cancer but could also be important in head and neck cancer.

Tobacco-related cancer in relation to prevalence of drinking and smoking in eastern Austria.

Eastern Austrian regional mortality patterns of oral cancer (oral cavity, pharynx and larynx) and oesophageal, lung and urinary bladder cancer were compared to smoker rates and to liver cirrhosis mortality by type of residence: Vienna (1.7 x 10(6) inhabitants), middle towns (50,000-100,000 and 10,000-50,000 inhabitants), small towns (2000-10,000 inhabitants) and rural areas categorized by agrarian quota less than or equal to 10%, 10%-20% and greater than 20%. The study area (Vienna, Lower Austria and Burgenland) covers 23,600 km2 with 3.23 x 10(6) inhabitants. In men, liver cirrhosis correlated negatively with smoker rates (r = 0.74, P = 0.1). Deaths from oral cancer and oesophageal cancer correlated significantly with deaths from liver cirrhosis (r = 0.81, P = 0.03; r = 0.78, P = 0.04, respectively) but not with smoker rates; lung cancer and bladder cancer correlated significantly with smoker rates (r = 0.91, P = 0.01; r0.83, P = 0.04, respectively), but not with liver cirrhosis. In women, similar urban-rural gradients for all parameters resulted in a positive correlation between liver cirrhosis and smoker rates (r = 0.59, P = 0.22) and a significant correlation of lung cancer with liver cirrhosis (r = 0.75, P = 0.05). Oral cancer correlated significantly with liver cirrhosis (r = 0.83, P = 0.02), but not with smoker rates; lung cancer correlated more significantly with smoker rates (r = 0.92, P = 0.01) than with liver cirrhosis; bladder cancer correlated positively with smoker rates (r = 0.70, P = 0.12). Geographical distribution of oral and oesophageal cancer in Eastern Austria seems thus to be highly subject to the prevalence of heavy drinking. Sociocultural influences upon the occurrence of these cancers seem to be mediated through drinking habits rather than through smoking habits alone.

Implication of membrane bound neuraminidase in developmental and adaptational processes of different vertebrate species.

Determination of brain membrane-bound sialidases by means of 4-methylumbelliferyl-neuraminic acid as exogenous, artificial substrate provides a good measure for functional implications of this enzyme in brain ontogeny and adaptation to changes in different environmental parameters. In rainbow trout and Cichlid fish a striking increase in brain sialidase-activity occurs during ontogenetical development, especially in post-hatching-larvae up to free swimming, reaching adult values of 75 and 50 nmol of sialic acid released per h and mg protein, respectively. Dark adapted Cichlid fish show a considerably lower turnover in brain (28 nmol per h and mg protein). Principally sialidase activity is in mammals about 5-fold lower than in fish and seems to be highest in cortex (compared with cerebellum and basal brain). Hibernating dwarf- and golden hamsters show a significant lower sialidase activity in summer, though these differences also occur in non-hibernating animals kept at constant ambient temperatures, thus indicating a temperature-independent, possibly genetically fixed enzyme regulation.

Aspergilloma in the frontal sinus expanding into the orbit.

A case of primary frontal sinus aspergilloma in a 79 year old non-immunocompromised woman, who presented with a right sided pyocele expanding into the orbit, is presented. The low susceptibility of the frontal sinus is probably related to the brachycephalic shape of the human skull which locates this sinus far anterosuperiorly to the nasal cavity. In human frontal sinus aspergillosis nasal symptoms are absent; the clinical manifestation of this rare disease is initiated by complications, especially orbital or intracranial invasion. Sinus opacity may raise early suspicion. Treatment consists of surgical debridement and re-aeration. Diagnosis is established by histological analysis of intraluminal contents.

A differential PCR system for the determination of CCND1 (cyclin D1) gene amplification in head and neck squamous cell carcinomas.

The gene for Cyclin D1 (CCND1) lies within chromosomal region 11q13 and codes for a cell cycle regulator essential for G1 phase progression. This G1-cyclin is a putative protooncogene whose clonal rearrangement and/or amplification and mRNA overexpression occurs in several types of human neoplasias, including head and neck squamous cell carcinomas. Data from the literature suggest that amplification and overexpression of the CCND1 gene could lead to destabilisation of cell cycle control mechanisms and uncontrolled cell proliferation. We developed a differential PCR system for the determination of CCND1 gene amplification in head and neck squamous cell carcinomas. A 115 bp CCND1 fragment and a 150 bp gamma-interferon fragment are amplified simultaneously in the same reaction tube under optimized conditions. Statistical analysis of amplification data obtained by differential PCR revealed excellent correlation with amplification data obtained by conventional Southern hybridization.

Cathepsin D and PAI-1 expression in human head and neck cancer.

In order to determine possible overexpression of cathepsin D and PAI-1 (plasminogen activator inhibitor) in head and neck tumours, cytosols from 92 primary squamous cell carcinomas (SCC), 19 lymph node metastases and 24 adjacent normal tissue samples were examined. For both cathepsin D and PAI-1, significantly elevated concentrations in SCC compared to normal tissue were found, even in pairwise comparison. Moreover, significantly higher cathepsin D and PAI-1 concentration in lymph node metastases than in corresponding normal tissue were also present. The conclusion was drawn that cathepsin D and PAI-1 may play a specific role in the biology of head and neck SCC.

Heightened embarrassability discriminates between panic disorder patients with and without agoraphobia.

Embarrassability refers to an individual's general susceptibility to becoming embarrassed and is closely linked to another personality characteristic known as fear of negative evaluation. To find out if panic disorder patients with and without agoraphobia differ in terms of embarrassability and fear of negative evaluation 100 patients with a DSM-III-R diagnosis of panic disorder with agoraphobia, 30 patients with a DSM-III-R diagnosis of uncomplicated panic disorder and 80 controls were administered the Embarrassability Scale and the 12-item version of the Fear of Negative Evaluation Scale. Depressive mood in the clinical group was assessed with the help of the Beck Depression Inventory. Comparisons between these three groups, between patients with mild, moderate, and severe phobic avoidance and between male and female subjects were carried out. Patients with agoraphobic avoidance showed significantly higher scores on both scales than patients with uncomplicated panic disorder and controls and women generally showed higher embarrassability scores than men. We conclude that heightened embarrassability is an important characteristic of patients suffering from panic disorder with agoraphobia.

Prognostic factors for swallowing rehabilitation following head and neck cancer surgery.

Thirty-two head and neck surgical patients with prolonged moderate to severe aspiration were assessed with videoendoscopic and videofluoroscopic swallowing studies to reveal all components of dysphagia and aspiration. All patients received functional swallowing therapy, and 75% of the patients regained full oral intake diet. The duration of non-oral feeding varied widely. The outcome of swallowing rehabilitation (success or failure, duration of non-oral feeding) was statistically correlated with preoperative tumour stage, patients' age, therapy onset, severity of aspiration and the results of the videofluoroscopic measurements of oral and pharyngeal transit time, pharyngeal delay time, duration of laryngeal closure and cricopharyngeal opening, hyoid and laryngeal elevation, presence or absence of a stenosis at the pharyngoesophageal segment. The following factors proved to be statistically significant for the prognostic estimate of swallowing rehabilitation: preoperative tumour stage, therapy onset, and severity of aspiration. For postoperative swallow recovery, an early therapy onset after thorough diagnostics with videoendoscopic and videofluoroscopic swallowing studies is recommended. Videofluoroscopic measurements will yield some prognostic estimate of oropharyngeal dysphagia and aspiration. Videoendoscopy, by it's availability and immediacy, proved to be useful for monitoring the course of rehabilitation.

Fine structure of the tendon of the human stapedius muscle: comparison of normals and clinically otosclerotic patients.

The tendon of the human stapedius muscle was studied in normal post mortem material and in clinical otosclerotic patients, using light and electron microscopy. Cross section profiles of collagen fibrils were measured in various regions of the tendon and the amount of elastin was estimated. The normal stapedius tendon consisted of three concentrically arranged portions: A cylindrical central part, a tube-like mid-portion, and a cortical layer. The central part was made up of collagen fibrils with only a few elastic fibers, the mid-portion contained collagen fibrils together with significantly more elastic material, while the cortical layer, again, showed a smaller amount of elastic fibers. Mean diameters of collagen fibrils in the central part of the tendon were 65.12 +/- 11.89 nm, in the intermediate layer 41.00 +/- 9.63 nm, and in the cortical layer 70.28 +/- 19.58 nm. Stapedius tendons from clinically otosclerotic patients, though showing the same construction, were characterized by significantly altered collagen fibrillar diameters (Mann-Whitney U-test). In the central part, mean diameters were reduced to 61.05 +/- 14.70 nm, in the mid-portion increased to 50.90 +/- 10.08 nm, and in the cortical layer reduced to 61.09 +/- 8.49 nm. The changes of collagen cross section profiles estimated for the entire tendon were significant as well: 59.68 +/- 18.74 nm in controls versus 57.82 +/- 12.53 nm in otosclerotic patients. Elastin content in the mid-portion of control stapedius tendons increased with age (13% at 35 years of age to 35% at 70 years of age).(ABSTRACT TRUNCATED AT 250 WORDS)

[Late functional outcome after reconstruction of the upper aerodigestive tract with free transplanted microvascular anastomosed jejunum]. / Funktionelle Spätergebnisse nach Wiederherstellung der oberen Atem- und Schluckwege mit frei transplantiertem mikrovaskulär anastomosiertem Jejunum.

Since 1983, 90 autologous jejunal transplantations for reconstruction of the upper digestive tract have been performed in 89 patients (9 females, 80 males, average age 56.3 years). 73 patients were operated primarily, in 16 patients a recurrent tumor had been treated. One patient received a second jejunal graft after necrosis. In these heterogenous patients, the primary tumor was located in the hypopharynx 48 times, in the larynx 21 times, in the oropharynx 19 times and twice in the oral cavity. There was nearly always tumor stage III or IV without distant metastases. Following locoregional tumor resection, speech restoration was achieved 35 times by a siphon-like tube, and the upper digestive tract was reconstructed using a patch 18 times and by a tube 28 times. A combination of tube or patch with a siphon-like tube was employed 9 times. In only two of 16 patients with recurrent tumor, speech reconstruction was performed, in the other 14, the upper digestive tract was reconstructed. In those patients, in whom speech reconstruction (by siphon tube or by combination with tube/patch) was intended, this was achieved in 81.5%. Successful functional reconstruction of the upper digestive tract (by patch, tube, combination) could be achieved in about 60% of these patients.

[A severe late tonsillar hemorrhage 2 months postoperatively]. / Eine schwere Tonsillenspätblutung zwei Monate postoperativ.

Tonsilloadenoidectomy performed on an 8-year old girl, in the course of which heavy bleeding from the left niche was staunched by suture ligation, was complicated from the 7th day onwards by severe haemorrhages at intervals from 5 to 25 days, causing life-threatening hypovolaemia. Haemorrhages started mostly in the early morning hours and ceased spontaneously. Revision of the left niche was undertaken shortly after the 5th bleeding two months postoperatively. After removal of the almost completely restituted mucosal covering, spreading of a haemosiderinstained canal released a massive pulsating haemorrhage which was controlled by ligation of the external carotid artery and suturing of the tonsillar bed. As the origin of the haemorrhage, an arterial lesion either by tonsil enucleation or by suture ligation is discussed. Attention is drawn to the possibility that certain branches of the external carotid artery may be positioned very close to the inferior half of the tonsil.

[The branchiogenic cyst in the oncologic context]. / Die branchiogene Zyste im onkologischen Zusammenhang.

8 patients with cystic carcinoma resembling a lateral cervical cyst by morphology and topography are reported. In 4 cases an analogous carcinoma later appeared in the ipsilateral tonsil, and in 2 tonsillectomized patients subsequent carcinoma developed in the palatopharyngeal arch or at the base of the tongue respectively. One carcinomatous cyst with consecutive involvement of the base of the tongue was preceded by a regular branchiogenic cyst on the opposite side. Removal of a recurrent branchiogenic cyst in another patient revealed unsuspected chronic myeloic leukaemia infiltrating adjacent lymph nodes. The hypotheses regarding the branchiogenic and lymphogenic pathogenesis of such carcinomatous cysts are discussed. Diagnosis should be elaborated as in any carcinomatous lump of the neck and should include serial sectioning of the tonsils and imaging of deep layers. The possibility of synchronous or metachronous tonsillar involvement must always be taken into consideration.

[Frequency dependence of the SISI test.(short increment sensitivity index). Clinical examples]. / Zur Frequenzabhängigkeit des SISI-Tests. Klinische Beispiele.

Experimental data on the influence of test frequency on the SISI-score are controversial. Comparability is limited by the diversity of experimental conditions. In a series of cases of pancochlear sudden hearing loss SISI-scores at 0.5 kHz and 3 kHz were compared. Some examples show that the sensitivity of the SISI-test at 0.5 kHz is much lower to that at 3 kHz. Low SISI-scores, of little evidence per se, have to be considered very critically in the low pitch range.

[Salivary gland tumors. Clinical aspects and therapy]. / Speicheldrüsentumoren. Klinik und Therapie.

Salivary gland tumors are rare. One-fifth are malignant. The parotid is the gland most often affected, particularly in adenomas. Adenomas and carcinomas are tumors that occur in adulthood; angiomas are the tumors that occur most frequently in childhood. Other than their localization, salivary gland tumors have few specific signs. In the clinical examination the locoregional and functional presentation is evaluated. Imaging studies refine or correct the clinical diagnosis, primarily by visualizing the deeper planes. Morphological diagnosis in benign lesions depends on resection results; in malignant lesions additional intraoperative frozen section is needed, and in questionable cases incisional biopsy. Fine-needle aspiration cytology may provide valuable knowledge when planning therapy. Limited parotid tumors are resected by lateral or subtotal parotidectomy. Carcinomas are resected as needed, with neck dissection added in case of lymph-node involvement. Small adenoid cystic carcinomas can be treated curatively by very wide resection. Aggressive and extensive tumors are irradiated postoperatively. Malignancies not operated upon are irradiated primarily. Facial nerve paralysis can be rehabilitated by different nerve repair or static surgical techniques.