Chrysotile fibers in tissue adjacent to laryngeal squamous cell carcinoma in cases with a history of occupational asbestos exposure.

Asbestos describes a group of naturally occurring fibrous silicate mineral compounds that have been associated with a number of respiratory maladies, including mesothelioma and lung cancer. In addition, based primarily on epidemiologic studies, asbestos has been implicated as a risk factor for laryngeal and pharyngeal squamous cell carcinoma (SCC). The main objective of this work was to strengthen existing evidence via empirical demonstration of persistent asbestos fibers embedded in the tissue surrounding laryngeal and pharyngeal SCC, thus providing a more definitive biological link between exposure and disease. Six human papillomavirus (HPV)-negative laryngeal (n = 4) and pharyngeal (n = 2) SCC cases with a history working in an asbestos-exposed occupation were selected from a large population-based case-control study of head and neck cancer. A laryngeal SCC case with no history of occupational asbestos exposure was included as a control. Tissue cores were obtained from adjacent nonneoplastic tissue in tumor blocks from the initial primary tumor resection, and mineral fiber analysis was performed using a scanning electron microscope equipped with an energy dispersive X-ray analyzer (EDXA). Chrysotile asbestos fiber bundles were identified in 3/6 of evaluated cases with a history of occupational asbestos exposure. All three cases had tumors originating in the larynx. In addition, a wollastonite fiber of unclear significance was identified one of the HPV-negative pharyngeal SCC cases. No mineral fibers were identified in adjacent tissue of the case without occupational exposure. The presence of asbestos fibers in the epithelial tissue surrounding laryngeal SCC in cases with a history of occupational asbestos exposure adds a key line of physical evidence implicating asbestos as an etiologic factor.

[Laryngeal paraganglioma]. / Paraganglioma gortani.

The paper describes a clinical case of laryngeal paraganglioma, a rare tumor. It provides the detailed characteristics of current diagnostic techniques.

Combinatorial effects of geopropolis produced by Melipona fasciculata Smith with anticancer drugs against human laryngeal epidermoid carcinoma (HEp-2) cells.

The identification of natural products exerting a combined effect with therapeutic agents could be an alternative for cancer treatment, reducing the concentration of the drugs and side effects. Geopropolis (Geo) is produced by some stingless bees from a mixture of vegetable resins, gland secretions of the bees and soil. It has been used popularly as an antiseptic agent and to treat respiratory diseases and dermatosis. To determine whether Geo enhances the anticancer effect of carboplatin, methotrexate and doxorubicin (DOX), human laryngeal epidermoid carcinoma (HEp-2) cells were treated with Geo alone or in combination with each drug. Cell growth, cytotoxicity and apoptosis were evaluated using 3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release, and flow cytometry. Scratch assay was used to analyze cell migration and transmission electron microscopy to observe morphologic alterations. The influence of Geo on drug resistance was also investigated assessing P-glycoprotein (P-gp) action. Geo inhibited cell proliferation and migration. The combination Geo+DOX led to the highest cytotoxic activity and induced apoptosis, leading to loss of membrane integrity. Geo had no effect on P-gp-mediated efflux of DOX. Data indicate that Geo combined with DOX could be a potential clinical chemotherapeutic approach for laryngeal cancer treatment.

[The electron microscopic study of tumour-like structures on the vocal cords].

We have undertaken the electron microscopic investigation into peculiarities of six tumour-like structures on the vocal cords. The study has demonstrated changes in the number and distribution patterns of intercellular junctions, keratin and tonofilament contents in epithelial cells, basal membrane structure, and composition of the basic substance in lamina propria. All the examined tumour-like structures contained bacteria an two of them had viral particles in vacuoles of fibroblasts. Moreover, the bacteria were found on the surface of epithelium, between epithelial cells and in the basic substance in lamina propria. Cytoplasm of epithelial cells and fibroblasts not infrequently contained bacteria in the phase of division.

Comparative ultrastructure analysis of radiation-induced radioresistant laryngeal cancer hep-2 cell line.

Radioresistance is one of the main reasons for the failure of radiotherapy in laryngeal cancer. However, the mechanisms of radioresistance of tumor cells have remained elusive. This study was conducted to identify the ultrastructural changes of radiation-induced radioresistant laryngeal cancer hep-2 cell line. First, a radioresistant hep-2R cell line was generated after prolonged exposure to γ-rays for 60 Gy (6 Gy/day, 2 days/week) and was confirmed by clonogenic assay. Next, the ultrastructural differences between hep-2R cells and hep-2 cells were compared by transmission electron microscopy. Finally, the results showed that hep-2R cells showed significant resistance to radiation compared with parental hep-2 cells. Increased cell nucleus atypia, more rough endoplasmic reticulum and less mitochondria were observed in hep-2R cells. The amount of microvilli of hep-2R was similar to hep-2 cell. In summary, these ultrastructural differences revealed the morphological mechanism that hep-2R cells had stronger radioresistance than hep-2 cells.

Evaluation of optical coherence tomography to discriminate lesions of the upper aerodigestive tract.

BACKGROUND: This study evaluated the capability of optical coherence tomography (OCT) to differentiate premalignant and early malignant lesions of the upper aerodigestive tract (UADT). METHODS: An OCT screening was performed in 52 healthy volunteers. Epithelial thicknesses on 38 OCT images and histopathological slides were correlated. One hundred primary lesions were rated via OCT concerning invasiveness by an "unblinded" investigator, then biopsied, and the results correlated. All OCT images were evaluated by 3 "blinded" investigators. Forty-eight images underwent retrospective image analysis. RESULTS: Screening showed large differences concerning epithelial thicknesses, but good correlation (κ = 0.63) between OCT and histopathological slides. In the unblinded evaluation, noninvasive and invasive lesions could be distinguished with a sensitivity of 88.9% and specificity of 89.0% whereas the blinded evaluations led to sensitivities of 100%, 66.7%, and 77.8% and specificities of 75.8%, 71.4%, and 70.3%. The difference of mean intraepithelial intensity reductions in dysplasias (38.7%) and hyperplasias (18.9%) was statistically significant. CONCLUSION: OCT complements visual inspection for differentiating UADT-lesions.

Managing chondrosarcoma of the epiglottis: a case report.

Laryngeal chondrosarcomas are a very rare malignancy with less than 150 cases reported in the literature. Of these, the epiglottis is the most unusual primary neoplastic subsite. Uncertainties arise owing to the extremely rare nature of the condition with regard to treatment and investigation for metastases in overtly low grade cases. We present the case of a 62-year-old woman with a low grade chondrosarcoma, arising from the tip of the epiglottis, presenting with dysphagia but no other symptoms.

[Effects of protein kinase CK2α on apoptosis and ultrastructure of laryngeal carcinoma cells].

OBJECTIVE: To investigate the effect of protein kinase CK2α on apoptosis and ultrastructure of human laryngeal carcinoma cells and its possible mechanism. METHODS: The siRNA expression plasmid psiRNA-hH1neo-CK2α specific to protein kinase CK2α and non-specific siRNA expression plasmid psiRNA-hH1neo-cont were transfected into Hep-2 cells respectively by lipofectamine method. Western blot was used to detect the expression of kinase CK2α protein. The apoptotic rate was measured by Annexin V-FITC/PI double-staining. The morphological changes of Hep-2 cells were observed under transmission electron microscope (TEM). The expressions of bcl-2 and Bax protein were measured by Western blot. RESULTS: The expression of protein kinase CK2α protein significantly decreased in the Hep-2 cells transfected with psiRNA-hH1neo-CK2α (P < 0.01). Compared with the untransfected cells and siRNA-hH1neo-cont transfected group, psiRNA-hH1neo-CK2α transfected group presented with classical ultrastructural features of apoptosis, such as karyopyknosis, chromatic agglutination adjacent to nuclear membrane and apoptotic body. The apoptotic rate of psiRNA-hH1neo-CK2α transfected group was obviously higher than that in untransfected cells and siRNA-hH1neo-cont transfected group (25.66% ± 0.83% vs 3.66% ± 0.43%, 5.18% ± 0.22%, both P < 0.05). Compared with two other groups, the bcl-2 protein expression of psiRNA-hH1neo-CK2α transfected group decreased (0.20 ± 0.09 vs 0.72 ± 0.16, 0.56 ± 0.11, both P < 0.01), the Bax protein expression increased (0.81 ± 0.17 vs 0.26 ± 0.12, 0.33 ± 0.17, both P < 0.01) while the ratio of bcl-2 to Bax decreased (0.25 ± 0.05 vs 2.76 ± 0.21, 1.70 ± 0.22, both P < 0.01). CONCLUSION: Protein kinase CK2a plays an important role in the apoptosis of human laryngeal carcinoma cells possibly by decreasing bcl-2/Bax. Protein kinase CK2a may provide a potential therapeutic target against human laryngeal carcinoma.

[The cytoskeleton and immigration of laryngeal squamous carcinoma cell affected by Fascin-1 expression].

This study sought to detect the effect of Fascin-1 expression on the cytoskeleton and immigration of laryngeal squamous carcinoma cell. In the experiment, Fascin-1 expression in Hep-2 cells was inhibited by small interfering RNA. The cytoskeleton of Hep-2 cells was observed with the use of laser scanning confocal fluorescence microscope. Millicell insert was applied to detect the immigration of Hep-2 cells in vitro. The results showed that the integrity of cytoskeleton in Hep-2 cells was broken with the down-regulation of Fascin-1 expression and the immigration ability was decreased significantly (P < 0.05). The inhibiting ratio of cell immigration was 44.6 +/- 6.3%. In conclusion, inhibition of Fascin-1 expression in Hep-2 cells could break the integrity of cytoskeleton and decrease the ability of cellular immigration.

Laryngeal papillomatosis: morphological study by light and electron microscopy of the HPV-6.

UNLABELLED: Laryngeal papillomatosis is the most frequent benign neoplasia in children. It is caused by HPV 6 and 11. The lesions are exophytic and highly recurrent, compromising the airway mucosa, mainly the larynx. Study design--clinical prospective. AIMS: To show morphologic alterations of the epithelium (light and electron microscopy) in the HPV-6 lesions. METHODS: Specimens of laryngeal lesions obtained during surgery of four children (1 male, 3 female) were submitted to HPV typing (PCR), light microscopy and electron microscopy. RESULTS: In all specimens, HPV type 6 was found. Epithelial projections were found by electron microscopy with superficial cells in desquamation. Light microscopy showed exophytic projections of the keratinized stratified squamous epithelium overlying a fibrovascular core. Koilocytes (vacuolated cells), suggesting the viral infection by HPV, were identified. No alterations were seen in the basement membrane and corion. Ultraestrutural analysis showed vacuolated cells with clear cytoplasmic inclusions, intercellular injuries and widening intercellular spaces. CONCLUSIONS: morphologic alterations of the epithelium in the HPV-6 lesions are superficial, and additional studies including the others HPV types are needed to show the more aggressive and extensive aspect of the disease.

Papilomatose laríngea: análise morfológica pela microscopia de luz e eletrônica do HPV-6 / Laryngeal papillomatosis: morphological study by light and electron microscopy of the HPV-6

Papilomatose laríngea é neoplasia benigna mais freqüente nas crianças, causada pelo HPV, principalmente subtipos 6 e 11 e caracteriza-se pela presença de lesões proliferativas exofíticas e recidivantes sobre a mucosa das vias aérea, em especial na laringe. Forma de Estudo: Clínico prospectivo. OBJETIVOS: Demonstrar alterações epiteliais morfológicas (pela microscopia de luz e eletrônica) em lesões papilíferas casadas pelo HPV-6. MATERIAL E MÉTODOS: Fragmentos de lesões de papilomatose laríngea, colhidos durante procedimento cirúrgico de quatro crianças (1 masculino, 3 femininas), foram submetidos à tipagem do HPV (por método de PCR), análise pela microscopia de luz e microscopia eletrônica (varredura e transmissão). RESULTADOS: Na tipagem, todos os papilomas eram do subtipo 6. A microscopia de varredura identificou projeções epiteliais de vários tamanhos, com células superficiais em descamação. A microscopia de luz demonstrou lesões exofíticas, revestidas por epitélio hiperplásico com coilócitos e binucleações, característicos do HPV. A membrana basal e o córion adjacente estavam íntegros. À microscopia eletrônica de transmissão identificou-se vacuolização perinuclear e alargamento das junções intercelulares. CONCLUSÕES: As alterações morfológicas apresentadas pelo HPV-6 demonstram o caráter não-invasivo da lesão, sendo necessário estudos morfológicos adicionais relacionando os outros tipos de HPV, considerados mais agressivos, com os achados ultra-estruturais.

Laryngeal papillomatosis is the most frequent benign neoplasia in children. It is caused by HPV 6 and 11. The lesions are exophytic and highly recurrent, compromising the airway mucosa, mainly the larynx. Study design - clinical prospective. AIMS: to show morphologic alterations of the epithelium (light and electron microscopy) in the HPV-6 lesions. METHODS: specimens of laryngeal lesions obtained during surgery of four children (1 male, 3 female) were submitted to HPV typing (PCR), light microscopy and electron microscopy. RESULTS: in all specimens, HPV type 6 was found. Epithelial projections were found by electron microscopy with superficial cells in desquamation. Light microscopy showed exophytic projections of the keratinized stratified squamous epithelium overlying a fibrovascular core. Koilocytes (vacuolated cells), suggesting the viral infection by HPV, were identified. No alterations were seen in the basement membrane and corion. Ultraestrutural analysis showed vacuolated cells with clear cytoplasmic inclusions, intercellular injuries and widening intercellular spaces. CONCLUSIONS: morphologic alterations of the epithelium in the HPV-6 lesions are superficial, and additional studies including the others HPV types are needed to show the more aggressive and extensive aspect of the disease.

Alveolar soft part sarcoma of the larynx: a case report of an unusual location with immunohistochemical and ultrastructural analyses.

BACKGROUND: Alveolar soft part sarcoma (ASPS) is a rare mesenchymal neoplasm of uncertain origin. In this article, we report a case of ASPS occurring in the larynx, an extremely rare location for this rather unusual tumor. METHODS AND RESULTS: The patient was a 34-year-old Japanese woman who requested an examination for hoarseness. The tumor showed a proliferation of large polygonal cells with periodic-acid-Schiff-positive diastase-resistant intracytoplasmic granules, arranged in an alveolar growth pattern. The cytoplasm of the tumor cells was eosinophilic. Tumor cells were positive for vimentin and titin. Nuclear immunoreactivity for TFE3 was observed, and the Ki-67 labeling index was 14.7%. Ultrastructurally, electron-dense rod-shaped crystals were infrequently observed in the cytoplasm. This case was finally diagnosed as ASPS of the larynx. CONCLUSION: We discuss the histogenesis and differential diagnosis of ASPS with immunohistochemical and ultrastructural findings. TFE3 immunohistochemistry was found to be a very useful marker for the diagnosis of ASPS.

[Laryngectomy as a salvage procedure of the patients with massive postradiation injury of the larynx]. / Laryngektomia jako postepowanie ratujace u chorych z nasilonym odczynem popromiennym po radioterapii raka krtani.

INTRODUCTION: Author discusses problems and treatment principles of patients with massive postradiation injury, who had laryngectomy procedure as a result of insufficience of the farmacological treatment. MATERIAL AND METHODS: There were 12 patients who were performed laryngectomy as a treatment of massive postradiation injury of the larynx in the period 1975-2005. We suspected presence of persistent neoplasm with postradiation changes. Seven laryngectomies were performed after confirmation of the neoplasm in 1-3 biopsies. Three patients were treated operatively without this confirmation in spite of two biopsies which were negatively, and two patients were treated in this way without biopsies. RESULTS: Two patients had tomour free postlaryngectomy specimens in the histopathological examinations, and among 10 others the reccurence of the tumour after radiotherapy was present during the post-laryngectomy histopathological examinations. In 7 cases this reccurence was proved with massive postradiation injury in endoscopic biopsies before laryngectomy. DISCUSSION: Author presents his own problems and presents methods of treatment of the patients with massive postradiation injury of the larynx described in literature.

Ultrastructural evaluation of 585-nm pulsed-dye laser-treated glottal dysplasia.

OBJECTIVE/HYPOTHESIS: To describe the ultrastructural changes occurring within pulsed-dye laser (PDL)-treated glottal tissues. STUDY DESIGN: Prospective. METHODS: Nine patients presenting with glottal dysplasia requiring biopsy to rule out microinvasive carcinoma were enrolled in this prospective study. At least two samples were obtained in each case: one from a PDL-treated area and another from a non-PDL-treated area (obtained from a nonphonatory region as an internal control). In some cases, a third sample was obtained from the junction between PDL- and non-PDL-treated areas. All samples were examined with light microscopy (H and E stain) and transmission electron microscopy. Observations were made of morphological changes within the epithelium, epithelial/ superficial lamina propria (SLP) junction, and the lamina propria of tissues treated with the PDL. Eight of nine patients were followed for a period of 9-25 months (mean, 18 months) with two recurrences that were retreated with awake-PDL and followed for an additional 8.3 and 9.5 months without recurrence. Vocal fold appearance returned to normal within 3-4 weeks posttreatment. RESULTS: Intraepithelial desmosome junctions were preferentially destroyed, and regional blood vessels were coagulated. The PDL consistently caused a separation of epithelial cells away from the basement membrane. CONCLUSIONS: The PDL allowed for both a surgical and a nonsurgical multimodality method for treatment of precancerous lesions with minimal effects on the SLP.

Large cell neuroendocrine carcinoma of the larynx: a case report and a review of the classification of this neoplasm.

This report describes a case of large cell neuroendocrine carcinoma (LCNEC) of the larynx. A 74 year old man who presented with otalgia underwent direct laryngoscopy with biopsy, which revealed an invasive poorly differentiated carcinoma. Laryngectomy with bilateral neck dissections revealed invasion of the pre-epiglottic space by the tumour, with metastases to bilateral lymph nodes (AJCC T3N2c). The tumour was characterised by large cells with vesicular chromatin and prominent nucleoli. The cells were arranged in organoid and trabecular patterns with a background of extensive necrosis and numerous mitotic figures. Immunohistochemical and ultrastructural analyses confirmed the neuroendocrine nature of the tumour. Metastatic disease was present in the liver, and the patient died within weeks of surgery. LCNEC carcinoma is a rare tumour of the larynx. Recognition at this site is essential so that proper patient management can be initiated.

Leiomyosarcoma of the larynx: critical analysis of the diagnostic role played by immunohistochemistry.

Laryngeal leiomyosarcoma (LMS) is definitely considered a rare occurrence: our exhaustive literature review disclosed to date only 45 cases (including the new case we have reported). The morphological diagnosis of laryngeal LMS may be problematic on conventional light microscopy especially in small laryngeal specimens. In the past, LMSs of the larynx were frequently confused with other spindle-cell malignant tumors. Nowadays, immunohistochemical investigations are considered necessary to distinguishing LMS from other spindle-cell tumors. On immunostaining, LMS is usually positive for muscle-specific actin and negative for cytokeratins and epithelial membrane antigen. In most cases, immunohistochemical study allows reliable diagnosis of LMS, but it can produce ambiguous or inconclusive results when the tumor cells lack specific immunohistochemical reactivity. In these controversial cases, LMS diagnosis has to be substantiated by electron microscopy. Distinguishing laryngeal LMS from other spindle-cell malignancies plays a role not only in academic interest. Confusion in the diagnosis of a spindle-cell laryngeal malignancy could result in inappropriate clinical management and inadequate treatment.

The nuclear localization of NFkappaB and p53 is positively correlated with HPV16 E7 level in laryngeal squamous cell carcinoma.

The interaction between the HPV (human papilloma virus) 16 E7 and other cell growth factors, such as p53 and NFkappaB in laryngeal cancer is not clearly understood. The aim of this study was to examine the expression of these three proteins in tumor and non-tumor laryngeal tissues from patients with laryngeal squamous cell carcinoma. These three proteins were dominantly expressed in the nucleus and their levels were higher in the tumor tissue than in the non-tumor tissue, although the comparison between the tumor and non-tumor tissues of p53 staining did not reach significance. The intensity of the nuclear stain of E7 and p53 was stronger than that of p65, a subunit of NFkappaB. Correlation analysis revealed that there was a positive relationship between the level of HPV16 E7 and the expression of p65. The correlation between E7 and p53 was also significant, although to a lesser degree. The finding of nuclear localization of p65 suggests that NFkappaB is constantly activated in the laryngeal cancer cells, whereas the sequestration of p53 in the nucleus may represent a mutated form of p53, which is probably inactivated by HPV16 oncoproteins. In conclusion, this study suggests that the nuclear localization of NFkappaB and p53 may play a role in the development of human laryngeal squamous cell carcinoma infected with HPV16.

[Evaluation of the ultrastructure of the vocal folds mucosa in patients with presbyphonia]. / Ocena ultrastruktury blony sluzowej faldów glosowych u pacjentów z presbyphonia.

Ultrastructure of the vocal folds mucosa was evaluated in 50 elderly patients. Study material included larynx specimens obtained from autopsy and postoperative material after the total laryngectomy due to the cancer of recessus piriformis with unchanged vocal folds. The ultrastructure of tunica mucosa was evaluated by means of the transmission electron microscopy with the use of Opton 900-PC microscope. In the control group the multilayer flat epithelium was found with the folded basal membrane, a large number of pericytes, scarce collagenous fibers in the stroma. The voice disturbances which occur during presbyphonia are conditioned by morphological changes in the epithelium, the basal membrane and the stroma of the vocal folds mucosa. Destruction of the epithelium cells with the enhanced vacuolar degeneration and enlarged intercellular spaces indicated oedemic character of presbyphonia. An increased number of collagenous fibers, vacuolar degeneration of fibroblasts with enlarged granular endoplasmic reticulum and an increased number of blood vessels in the stroma suggested an atrophic form of presbyphonia.