Is the carbohydrate sialosyl-Tn a marker for altered,non-malignant activity in squamous epithelium in the head and neck region?

Cell surface carbohydrates are involved in many cell functions such as cellular differentiation, adhesion, and invasion. A carbohydrate, sialosyl-Tn (STn), is expressed in many human carcinomas but generally not in normal epithelia. In the oral mucosa, however, STn had recently been observed on basal cell in some lesions with epithelial hyperplasia and dysplasia. The aim of this study was to carry out a systematic investigation of STn expression on epithelial basal cells in hyperplastic, ‘borderline’ malignant, and malignant head and neck lesions, to see if the expression of STn is associated specifically with hyperplastic conditions. Using the primary monoclonal antibody TKH2, normal controls did not reveal STn. STn was detected on probably post-mitotic basal cells in hyperplastic head and neck lesions and on basal cells adjacent to cancers, but not within the carcinomas. A Ki67 antibody reacted with basal cells in other locations. The most highly differentiated lesions, such as focal epithelial hyperplasia and verrucous hyperplasia, revealed a high percentage (86 per cent in both cases) of STn reactivity. The least-differentiated verrucous carcinomas (VCs) and keratoacanthomas (KAs) did not express STn, in contrast to the highly differentiated VCs and KAs. These findings indicate that STn-negative cases may have a greater malignant potential that the STn-positive cases. In conclusion, STn expressed on basal cells is possibly a marker for non-malignant conditions with altered basal cell activity and for highly differentiated verrucous carcinomas.     

Identification of Histoplasma capsulatum from culture extracts by real-time PCR

We designed and tested a real-time LightCycler PCR assay for Histoplasma capsulatum that correctly identified the 34 H. capsulatum isolates in a battery of 107 fungal isolates tested and also detected H. capsulatum in clinical specimens from three patients that were culture positive for this organism.     

Prognostic value of histobiological factors (malignancy grading and AgNOR content) assessed at the invasive tumour front of oral squamous cell carcinomas.

Tumour cells at the invasive front of carcinomas have been found to differ substantially from the rest of tumour cells in a variety of human cancers. The present multivariate survival analysis of 94 oral squamous cell carcinomas (OSCCs) revealed that both the argyrophilic nucleolar organizer regions-associated protein (AgNOR) content of invading tumour cells and a multiparametric histopathological tumour front grade were significantly and independently associated with tumour-related death, irrespective of conventional Broders'' grade and clinical stage of the tumours. High tumour front scores and AgNOR content at the invasive OSCC front thus seem to reflect increased malignant potential. Proliferative activity, assessed by standardized AgNOR analysis, most probably represents one of the biological features underlying the usefulness of evaluating the invasive tumour front.     

Detection of malignant epithelial cells in effusions using flow cytometric immunophenotyping: an analysis of 92 cases

We compared the efficiency of immunophenotyping using flow cytometry (FCM) and a combination of morphologic and immunocytochemical studies for detecting malignant cells in 92 effusions. Cytologic results were as follows: carcinoma cells, 43 specimens; benign, 42 specimens; suggestive of nonepithelial malignancy, 7 specimens. After immunocytochemical analysis, 5 benign specimens were reclassified as malignant and 4 malignant epithelial specimens as benign. With FCM, cells positive for Ber-EP4, B 72.3, AH6, and HB-TN were detected in 28 to 36 (64%-82%) of 44 carcinomas but only 2 to 12 (5%-29%) of 41 benign specimens. Significant association was seen for coexpression. Ber-EP4 and AH6 were the most sensitive; Ber-EP4 was the most specific. The presence of cells positive for 3 of 4 markers strongly suggested malignancy (34/44 carcinoma specimens [77%]; 3/41 reactive specimens [7%]). The presence of cells positive for all 4 markers was diagnostic of malignancy (17/44 malignant specimens [39%]; 0/41 reactive effusions [0%]). FCM and immunocytochemical resultsfor Ber-EP4 expression showed excellent association. FCM is a powerful tool for diagnosing difficult effusions and can quantify coexpression of various markers in fresh specimens. By using established cellular markers coupled with biological markers, FCM also has great promise for experimental purposes.     

Expression of cell cycle proteins in ovarian carcinoma cells in serous effusions-biological and prognostic implications

OBJECTIVE: The aim of this study was to investigate the expression of cell cycle proteins in ovarian carcinoma cells in serous effusions and respective solid tumors. METHODS: Fifty-five malignant effusions and 38 tumors (20 primary, 18 metastatic) were immunohistochemically stained for cyclin A, p27(kip1), and Ki-67. Staining extent (0-100% cells) and intensity (0-3 scale) were scored. Cyclin A and p27(kip1) expression was additionally studied in 29 malignant effusions using immunoblotting. Immunohistochemistry results in effusions were evaluated for possible association with clinicopathologic parameters. RESULTS: Nuclear immunoreactivity for all markers was detected on carcinoma cells in the majority of effusions using immunohistochemistry. Similarly, immunoblotting showed the presence of cyclin A and p27(kip1) in 29/29 and 25/29 specimens, respectively. Intense (3) immunoreactivity for Ki-67 was detected more often in peritoneal effusions, compared with those of pleural location (P = 0.036). Staining in primary and metastatic lesions was generally comparable to that of tumor cells in effusions. Staining for p27(kip1) was more diffuse in effusion specimens obtained prior to the institution of chemotherapy (P = 0.042). In an analysis of all effusions, an association was observed between the number of cells that were immunoreactive for Ki-67, cyclin A, and p27(kip1) (cyclin A-Ki-67: P = 0.008; p27(kip1)-Ki-67: P = 0.019; cyclin A-p27(kip1): P = 0.032). In survival analysis, the presence of more diffuse (P = 0.042) and intense (P = 0.019) staining for cyclin A correlated with prolonged overall survival. CONCLUSIONS: The expression of the studied cell cycle markers does not differ markedly between ovarian carcinoma cells in the pleural and peritoneal cavity, supporting our previous studies of several metastasis-associated molecules. The presence of cyclin-A-positive cell populations is associated with a more favorable disease outcome, possibly due to the targeting of proliferating cells by chemotherapeutic agents. However, the decline in the fraction of p27(kip1)-positive cells in posttreatment specimens may point to additional mechanisms involved in this selection.     

E-Cadherin complex protein expression and survival in ovarian carcinoma

OBJECTIVE:The aim of this study was to analyze the correlation between expression of E-cadherin complex proteins, epidermal growth factor receptor (EGFR), and c-erbB-2 and disease outcome in advanced-stage ovarian carcinomas. METHODS: Sections from 75 primary ovarian carcinomas (=37) and metastatic lesions (=38) from 45 patients diagnosed with advanced-stage ovarian carcinoma (FIGO stage III-IV) were immunostained and evaluated for staining pattern, extent, and intensity. Patients were divided in two groups based on disease outcome. Long-term survivors (21 patients) and short-term survivors (24 patients) were defined using a double cutoff of 36 months for disease-free survival (DFS) and 60 months for overall survival (OS). Mean follow-up period was 70 months. The mean values for DFS and OS were 109 and 125 months for long-term survivors, as compared to 3 and 21 months for short-term survivors, respectively. RESULTS: Comparison of all primary and metastatic lesions showed upregulation of gamma-catenin protein expression in the latter (P = 0.05). When segregated according to disease outcome, the expression of all studied proteins, with the exception of EGFR, was more diffuse in tumors of short-term survivors. The presence of cytoplasmic staining for c-erbB-2 was associated with poor survival in the entire cohort (P = 0.007), as well as in primary tumors alone (P = 0.003), in survival analysis. Similar results were seen in the evaluation of primary tumors for gamma-catenin (P = 0.002). CONCLUSIONS: gamma-Catenin, and possibly c-erbB-2, are valid markers of poor survival in advanced-stage ovarian carcinoma.     

Methods for animal satellite cell culture under a variety of conditions

Primary and clonal culture systems have been devised and refined for animal-derived satellite cells. Satellite cell (SC) culture development includes efficient cell isolation techniques, establishment of effective plating and growth conditions, formulation of media requirements and thorough evaluation of experimental limitations. As the field of muscle cell culture has expanded, the number of animal species from which satellite cells have been isolated has increased. The focus of this paper is to compare and contrast SC culture conditions presently used by a variety of researchers and to introduce a new source of SC from wapiti (elk).     

A study of phosphorylated ERK1/2 and COX‐2 in early stage (T1–T2) oral squamous cell carcinomas

Background: Histomorphological grading at the invasive front of oral squamous cell carcinomas (OSCCs) may provide useful prognostic information. In the present study, we investigated the presence and prognostic value of activated phosphorylated extracellular signal‐regulated kinases 1 and 2 (p‐ERK1/2) and cyclo‐oxygenase‐2 (COX‐2) both at the invasive front and in central/superficial parts of OSCCs. Methods: Using immunohistochemistry, we assessed the presence of p‐ERK1/2 and COX‐2 in 53 early stage OSCCs. Clinical data were recorded prospectively. The end point was disease‐free survival. Results: p‐ERK1/2 staining was present in almost all tumours. The staining was mostly nuclear in the cells of the invasive front and either nuclear or nuclear/cytoplasmic in central/superficial tumour parts. COX‐2 was observed in almost all tumours (98%) and the staining was often restricted to focal areas. Most tumours were COX‐2 negative at the invasive front. The lowest P‐value in survival analyses was P = 0.06 for p‐ERK1/2 at the invasive front. COX‐2, the histomorphological grading systems and TNM stage were of no prognostic value. Conclusion: p‐ERK1/2 was present in almost all tumours and p‐ERK1/2 may be a prognostic marker at the invasive front of OSCCs. In early stage OSCCs, most tumours did not express COX‐2 at the invasive front.     

Prognostic value of Rhesus blood groups in oral squamous cell carcinomas

In the current study of the prognosis of all patients (N equals 70) with squamous cell carcinomas (SCC) of floor of mouth in Norway during the period 1963 to 1972, the authors found that patients with Rhesus (Rh) (D)-negative blood group had significantly poorer prognosis (mean 5-year survival, 8%) than patients with Rh (D)-positive blood group (5-year survival, 30%) (P equals 0.04). This extends the authors' previous observations in another group of oral cancer patients. The authors do not know the explanation for this association. However, the Rh gene locus is located on the short arm of chromosome 1 which reportedly has shown rearrangements in some head and neck SCC and other human neoplasms. The authors therefore speculate that the Rh gene locus may be linked with chromosome 1 changes of importance for the progression of oral SCC.