Detection of HPV DNA in oral carcinoma using polymerase chain reaction together with in situ hybridization.

This study determined the prevalence of human papillomavirus 16/18 DNA in deparaffinized oral carcinoma specimens on slides with the use of the different sensitivities of in situ hybridization and a technique that combines polymerase chain reaction and in situ hybridization. Human papillomavirus DNA was not detected in the 30 biopsy specimens analyzed by in situ hybridization alone using biotinylated DNA probes specific for human papillomavirus 16/18. Twenty of 30 specimens (66.7%) were found to have human papillomavirus DNA (p < 0.001) with the use of the polymerase chain reaction-in situ hybridization technique. Human papillomavirus 16 was detected in 18 of 26 specimens (69.2%), and 7 of 25 carcinomas (28%) were found to contain human papillomavirus 18. Dual infections were present in 5 of 21 (23.8%) specimens. Human papillomavirus DNA was more prevalent in men (75%) than women (57.1%). However, there was no difference in the mean age of patients with oral carcinoma (men, 67.8 years; women, 67.5 years) who had human papillomavirus and those who did not (67.2 years). Carcinomas associated with dual infections occurred at a lower mean age (59.4 years) than those associated with a single human papillomavirus type (p < 0.005). We conclude that the polymerase chain reaction-in situ hybridization technique enhances our ability to demonstrate human papillomavirus types highly associated with oral squamous cell carcinoma.

In situ detection of HPV DNA in oral mucosal lesions. A comparison of two hybridization kits.

The purpose of this study was to compare the sensitivity of ViraType in situ hybridization kit (Life Technologies, Inc. [LT] and PathoGene (Enzo Diagnostics, Inc. [ED]) in situ hybridization kit for human papillomavirus (HPV) DNA detection in oral tissue. Forty benign oral lesions histologically suspicious for HPV infection were analyzed. Specimens were hybridized with DNA probes specific for HPV types 6/11, 16/18, and 31/33/35 [LT] and HPV types 6/11, 16/18, and 31/33/51 [ED]. Positive hybridization reactions were seen for HPV DNA type 6/11 only. Hybridization occurred significantly more often (p less than 0.01, McNemar Exact Test) in LT probed specimens (20/40) than ED assayed sections (12/40). HPV DNA sequences were found in 100% condyloma acuminata (13/13), 100% verruca vulgaris (4/4), and 13% squamous papilloma (3/23) using the LT system. The ED probes yielded positive signals in 77% condyloma acuminata (10/13), 25% verruca vulgaris (1/4), and 4.4% squamous papilloma (1/23). A more intense hybridization signal was exhibited using the LT system. The results indicate that the LT probes and detection reagents are more sensitive for detecting HPV DNA in oral mucosal specimens.

In situ hybridization analysis of human papillomavirus DNA in oral mucosal lesions.

Commercial biotinylated DNA probes specific for human papillomavirus (HPV) types 6 and 11; 16 and 18; and 31, 33, and 35 were used for in situ hybridization analysis of 105 oral mucosal specimens from 5 cases of verruca vulgaris, 15 cases of condyloma acuminatum, 30 cases of squamous papilloma, 20 cases of hyperkeratosis/acanthosis, 15 cases of epithelial dysplasia, 5 cases of carcinoma in situ, and 15 cases of squamous cell carcinoma. Positive hybridization signals were found in 26 specimens (24.8%). Only HPV-6/11 was detected. HPV DNA occurred significantly more often (p less than 0.005, chi-square analysis) in condyloma acuminatum (100%) and verruca vulgaris (100%) than squamous papilloma (13.3%), hyperkeratotic/acanthotic lesions (10%), and malignant and premalignant lesions (0%). The tongue (19.1%) and labial epithelium (17.1%) were infected most frequently. Nuclear reaction products indicating HPV infection were associated primarily with koilocytes. These results demonstrate the usefulness of commercial biotinylated probes for HPV DNA analysis in routine paraffin-embedded lesion specimens. They confirm HPV involvement in benign lesions of the oral mucosa but fail to associate HPV infection with oral cancer and precancer.

Acute effect of calcium channel blockers on adriamycin exposed isolated adult cardiocytes.

When tested with isolated, calcium-resistant resting rat cardiocytes in an in vitro assay system, adriamycin exerted a dose-dependent cytotoxic effect which could easily be assessed by the ATP depletion of the heart cells and the loss of vitality as monitored by morphological changes (blebbing, spherical contraction). Apart from extremely high non pharmacological concentrations of verapamil and diltiazem, both calcium antagonists left the cardiocytes intact and without loss of internal ATP when given alone to the medium. Coincubation of adriamycin and verapamil or diltiazem did not increase adriamycin toxicity to the cardiocytes; instead a remarkable ATP preservation by verapamil could be demonstrated when both drugs (adriamycin and verapamil) were incubated simultaneously with the heart cells. This acute protective effect was limited in time and could no longer be detected after 9 hours. Diltiazem in coincubation experiments exerted neither a toxic nor an acute protective effect on adriamycin-exposed heart cells.