Lack of type-specific concordance between human papillomavirus (HPV) serology and HPV DNA detection in the uterine cervix and oral mucosa.

There is limited knowledge about longitudinal genotype-specific concordance between human papillomavirus (HPV) serology and co-existent presence of HPV DNA in the uterine cervix. The role of oral HPV infections in inducing serological response is unclear, as is the effect of HPV antibodies on the outcome of oral HPV infections. The present study is part of the Finnish Family HPV Study designed to evaluate dynamics of HPV infections within families. Here, we correlated the point prevalence of HPV6, 11, 16, 18 and 45 antibodies and concomitant genotype-specific HPV DNA detection in cervical and oral samples of 323 mothers during their 3 year (mean 37.5 months) follow-up. The mean age of these pregnant mothers at enrolment (third trimester) was 25.5 years. HPV antibodies were analysed with multiplex HPV serology and HPV genotyping was performed using a Multimetrix kit (Progen Biotechnik). There was no concordance between cervical DNA detection and co-existent seropositivity, and the same was true even in samples taken 12 months apart. Women who cleared their cervical HPV16 infection had the highest HPV16 antibody levels, whereas those who acquired incident HPV16 infections had the lowest antibody levels. Neither the presence nor the dynamics of oral HPV DNA had any correlation with HPV serology.

Fas/CD95 promoter polymorphism gene and its relationship with cervical carcinoma.

OBJECTIVE: Apoptosis is an important fail-safe control in human papillomavirus (HPV)-associated carcinogenesis. We tested the hypothesis that the A/G polymorphism at -670 of Fas promoter is associated with an increased risk for cervical cancer, using a matched case-control setting. METHODS: The material in this case-control study consisted of 91 patients with cervical carcinoma and 176 population-based control subjects, recruited between 2002 and 2004; all the ethnic Brazilian women had histologically confirmed cervical carcinoma. Control subjects were age-matched; healthy women who were selected following a negative cervical cytology and normal colposcopy. Fas genotyping was performed using a PCR-RFLP technique. RESULTS: No significant difference existed in the distribution of the Fas polymorphisms (wild, heterozygous, mutant) between the cases and controls. The heterozygous (OR: 4.85, 95% CI: 1.1-22.6) genotypes among the younger (< 48 yrs) cancer patients were almost 5-fold increased, as compared with the wild type. No such increase was observed among the patients older than 48 years. CONCLUSIONS: Our data suggest that 670A/G polymorphism in the promoter region of the death receptor Fas is associated with an increased risk of cervical cancer among Brazilian women under 48 years. The mechanisms would be the inhibition of apoptosis by Fas -670G allele-mediated down-regulation of Fas transcription.

Early integration of high copy HPV16 detectable in women with normal and low grade cervical cytology and histology.

BACKGROUND: Integration of human papillomavirus (HPV) DNA has been considered a late event in cervical carcinogenesis. However, integrated forms of HPV were recently detected in cancer precursor lesions using a new real time polymerase chain reaction (PCR) to detect the deletions at the 3362-3443 region of HPV16 E2 OBJECTIVE: To study the frequency of HPV16 DNA integration in cervical lesions and compare the sensitivity of an additional upstream region of the E2 ORF (2962-3138) in detecting HPV integration. METHODS: Using the TaqMan based PCR, HPV16 positive DNA samples were analysed in 164 cervical scrapings from women participating in a multicentre screening trial. Biopsy confirmation was available in 62 cases. RESULTS: Primers targeting the 3362-3443 region detected the majority of E2 deletions. In only 23% of the samples was the E2 upstream region equal or better target than the 3362-3443 region. Mixed (episomal/integrated) pattern was the most prevalent physical state of HPV16, also present in PAP smears with normal morphology. Pure integrated form was most prevalent in HSIL and cancer lesions, but also detectable in low grade abnormalities (NSIL, ASC-US, LSIL). Women with only integrated HPV16 were almost 10 years older than those with episomal HPV16. Viral load of integrated HPV16 was related to cytological abnormality (p = 0.003) but not to histology. CONCLUSIONS: Integrated HPV16 is present in low grade cervical lesions, mostly mixed with the episomal form. Women with the pure integrated form of HPV16 are older than those with the other forms.

High levels of stromal hyaluronan predict poor disease outcome in epithelial ovarian cancer.

Several malignant tumors accumulate hyaluronan, a matrix component suggested to promote cancer cell migration and growth. To explore the potential clinical importance of this concept, we assessed the hyaluronan levels in epithelial ovarian cancer. A biotinylated affinity probe specific for hyaluronan was prepared and applied to histological sections of 309 epithelial ovarian cancers and 45 matched metastatic lesions. The staining was scored according to the percentage area of strong hyaluronan signal of total peri- and intratumoral stroma as low (<35%), moderate (35-75%), or high (>75%). Low, moderate, and high levels of stromal hyaluronan were observed in 95, 116, and 98 carcinomas, respectively. The high stromal hyaluronan level was significantly associated with poor differentiation, serous histological type, advanced stage, and large primary residual tumor, whereas it was not correlated with high CD44 expression on cancer cells. The 5-year outlook of the disease deteriorated with increasing stromal hyaluronan levels for both overall (45% versus 39% versus 26%; P = 0.002) and recurrence-free (66% versus 56% versus 40%; P = 0.008) survival. High levels of stromal hyaluronan were more frequent in metastatic lesions than in primary tumors (z = -3.9; P = 0.0001). In Cox's multivariate analyses, high level of stromal hyaluronan was an independent prognostic factor in all patients, as well as in stage-specific subgroups. These results suggest that stromal hyaluronan accumulation may be a powerful enhancer of tumor progression and, as such, provides a novel, independent prognostic marker and a potential target of therapy.

Clearance of high-risk human papillomavirus (HPV) DNA and PAP smear abnormalities in a cohort of women subjected to HPV screening in the New Independent States of the former Soviet Union (the NIS cohort study).

BACKGROUND: We analysed the temporal relationships of the clearance of human papillomavirus (HPV) DNA and cytological abnormalities in women participating in a screening study in three NIS countries. METHODS: The 274 patients included in this analysis were prospectively followed-up for 21.6 months (range: 0.5-42.9). All 274 women had abnormal PAP test (ASC-US or higher) and high-risk HPV-positive test (HCII) at baseline. Two groups were compared: 132 women who cleared both tests (Group 1), and 142 women who cleared either HPV or abnormal PAP test (Group 2). The first clearance during the follow-up, and the last visit clearance were modeled using life-table techniques, and the predictive factors were analysed using univariate (Kaplan-Meier) and multivariate (Cox) survival analysis. RESULTS: There was no difference in the mean clearance time for the abnormal PAP test (14.4 months; 0.7-40.5 and 12.6 months; 0.5-35.0) and high-risk HPV DNA (12.67 months; 0.6-33.5 and 10.8 months; 0.7-33.4) in Group 1 and Group 2 (Mann-Whitney: P = 0.107 and P = 0.082, respectively). Clearance times for HPV DNA and abnormal PAP test did not deviate from each other in either groups (Wilcoxon: P = 0.063 and P = 0.088). The monthly clearance rates for the abnormal PAP test are 1.32 and 1.38%, and those for the HPV DNA 1.62 and 1.61%, in Groups 1 and 2, respectively. Of the factors predicting the last visit clearance, the issues related to smoking are of particular interest. CONCLUSIONS: The clearance of high-risk HPV type and abnormal PAP test shows a close temporal relationship, the former preceding the latter, however, by an interval of 1.0-2.0 months.

Immunohistochemistry in assessment of molecular pathogenesis of cervical carcinogenesis.

Concerning the prediction of HPV-associated cervical disease, several importance issues are related both to the management of women with diagnosed CIN and those with cervical cancer. Oncogenic HPVs are capable of contributing to the development of malignant phenotype by several different mechanisms, most of which seem to be closely interrelated. Because of the fact that these molecular interactions are mediated by proteins, the logical strategy to dissect the complex molecular pathways is to study the functions of these proteins, utilising the capabilities of immunohistochemistry (IHC). IHC offers practically unlimited possibilities to study any target molecules, against which a monoclonal or polyclonal antibody can be raised. This review describes the IHC-based strategies used by this author to assess the molecular pathogenesis of cervical cancer and its precursors in a number of large-scale prospective cohort studies conducted during the past 25 years. In the ongoing HPV-PathogenISS study, 13 different markers are being tested to evaluate their predictive value in distinct viral events, e.g. persistence or clearance of high-risk HPV in women treated for CIN. Apart from getting new insights into the molecular pathogenesis of HPV-associated cervical carcinogenesis, we anticipate the disclosure of individual markers, a set of markers, or an expression profile of any such marker sets that would be of clinical value as predictors of disease outcome in cervical carcinogenesis.

The role of nucleolar organiser regions as prognostic factors in breast cancer.

Nucleolar organiser regions (NORs) were stained in paraffin-embedded biopsy specimens of 80 female breast carcinomas by the silver (Ag) technique. The patients were prospectively followed up for a mean of 12.4 years (range 11.5-13.3). The number of different types of Ag-NORs was correlated with the histological grade, clinical stage, DNA ploidy, S-phase fraction (SPF) and clinical outcome. Grade III tumours showed higher Ag-NOR counts than low grade tumours. The total number of Ag-NORs (P = 0.0059) and the number of dispersed Ag-NOR (P = 0.0199) were significantly related to DNA ploidy aneuploid tumours showing higher Ag-NOR counts. The number of aggregated Ag-NORs was predictive (P = 0.0413) for the development of metastatic disease during follow-up. On the other hand, crude, cancer-related or recurrence-free survival could not be predicted significantly by the Ag-NORs. The results suggest that the number of Ag-NORs is clearly related to the proliferative activity in breast cancer, but the prognostic value of Ag-NOR counting is inferior to the previously recognised prognostic factors.

Morphology of the spleen white pulp in relation to the immunological functions in patients with far advanced bronchial carcinoma.

The white pulp of the spleens collected from seventy-nine patients died in far advanced bronchial carcinoma, and the same number of spleens from an age-matched group of patients died in myocardial infarction without evidence of any malignancy was histologically assessed by using a standardized reporting system recently introduced by the author. Emphasis was placed on the evaluation of the cell populations involved in immunological reactions. Histological characteristics suggesting an active function of both the cell-mediated and humoral immune reactions (T- and B-cell areas) were found to be within normal limits in the control series, but profoundly deranged in the cancer patients. Active germinal centers were infrequently seen in both series. The significance of the histological observations made was discussed in the light of the impaired immunological reactivity associated with human malignancies, and a conclusion was drawn, although with some caution, that such an impairment of both the humoral and cell-mediated immune responses might probably exist in a considerable number of the present patients dying of far advanced bronchial carcinoma. The applicability of the reporting system now used for the first time in human pathology was emphasized.

Epidemiology of human papillomavirus (HPV) infections and their associations with genital squamous cell cancer. Review article.

Reliable assessment of the epidemiology of genital HPV infections is hamphered by a number of technical problems. Because of the lack of tissue-culture systems, methods based on morphological approaches (colposcopy, cytology and histopathology) play a central role in HPV diagnosis. Even DNA-hybridization techniques and the recently introduced DNA amplification with PCR are extremely difficult to standardize, and are thus subject to major interlaboratory variation. Further confusion in the field is created by the complex biological behaviour of HPV infections. As established by the long-term prospective follow-up study of over 500 women which has been running in Kuopio since 1981, clinical progression and regression are significantly related to the grade of the lesion at the time of diagnosis (p less than 0.00001, and p = 0.0005, respectively), as well as to the type of HPV (p = 0.0012). Most importantly, however, genital HPV infections seem to run an extremely fluctuating course, passage from manifest to subclinical or latent infection being frequently encountered in individual patients when examined at 6-month intervals over prolonged periods. This explains the significantly divergent prevalence figures reported in different series (ranging from 2% to 80%), which are completely dependent on the technique used to analyse the presence of HPV, i.e. whether a) PAP smear, b) biopsy, c) DNA hybridization, or d) PCR amplification. The first two are capable of disclosing only manifest (clinical) infections, the latter two also the latent ones. In an unselected population of 22-year-old Finnish females, the prevalence of clinical HPV infections was about 3 per cent, and the adjusted annual incidence was 8.0 per cent. According to estimates of the life-time risk, up to 79% of Finnish females will contract at least one HPV infection between the ages 20 to 79 years. When related to the long-term trends in invasive cervical cancer in Finland, it is evident that this 79% life-time risk of becoming HPV-infected or even the observed 15% clinical progression rate for HPV infections in the prospective follow-up study by no means signifies an identical risk of developing cervical cancer (i.e. 0.79 x 0.15 = 11%). It seems likely that in countries where mass-screening programmes exist (and precancer lesions are traced), the high prevalence of HPV infections is not necessarily reflected as an increased prevalence of invasive cervical carcinomas. The distinction of lesions at risk for malignant transformation from those regressing spontaneously will have major implications in therapeutic considerations of genital HPV infections.

Fine needle aspiration cytology of the thyroid osteosarcoma. Report of a case.

The fine needle aspiration cytology of a histologically verified primary osteosarcoma of the thyroid gland is reported for the first time in literature. The histogenesis and the differential diagnostic aspects of this extremely rare tumor are briefly discussed.

HPV infections in benign and malignant sinonasal lesions.

This review updates the evidence that the human papillomavirus (HPV) is involved in the development of benign and malignant sinonasal lesions. Since the early 1980s, when evidence was provided on the possible involvement of HPV in the aetiology of both benign respiratory papillomas and squamous cell carcinomas, a substantial number of studies have explored this issue. To date, 33.3% of sinonasal papillomas and 21.7% of sinonasal carcinomas analysed have been shown to be positive for HPV. Many elements of the data parallel the observations made in HPV lesions at other mucosal sites, such as malignant transformation and frequent recurrence after radical treatment; the fact that low risk HPV types 6 and 11 are usually confined to benign lesions, whereas the reverse is true for the oncogenic HPV types 16 and 18; and the presence of squamo-columnar junctions and squamous cell metaplasia in the sinonasal system. The discrepancies reported by several studies might result in part from technical reasons, but it is also possible that sinonasal lesions have a heterogeneous aetiology (HPV related and non-related) and/or that some novel (yet unidentified) HPV types exist in these lesions, which are detected by some studies but not by others.

HPV infections and oesophageal cancer.

The first reports suggesting an involvement of human papillomavirus (HPV) in the development of both benign and malignant squamous cell tumours of the oesophagus date back to 1982. Since then, a substantial amount of literature has accumulated on this subject, summarised in this review. To date, 239 oesophageal squamous cell papillomas have been analysed in 29 separate studies using different HPV detection methods, with HPV being detected in 51 (21.3%) cases. Many more squamous cell carcinomas have been analysed: of the 1485 squamous cell carcinomas analysed by in situ hybridisation, 22.9% were positive for HPV DNA, as were 15.2% of the 2020 cases tested by the polymerase chain reaction. In addition, evidence derived from large scale serological studies, animal experiments, and in vitro studies is discussed in the light of the highly variable geographical incidence rates of oesophageal carcinoma worldwide. It may be that the (multifactorial) aetiology of oesophageal cancer differs greatly between those geographical areas with a low risk and those with a high risk for this disease. Oncogenic HPV types seem to play an important causal role, particularly in high risk areas.

HPV infections and lung cancer.

Because of the major clinical impact of bronchial cancer worldwide, the possibility that human papillomavirus (HPV) contributes to its pathogenesis as a co-carcinogen is an intriguing one. Bronchial squamous cell carcinoma develops through well defined precursor lesions, often at the sites of squamous metaplasia. Benign squamous cell papillomas are rare but HPV DNA has been found in almost half of those studied, implicating a causal association. In invasive bronchial cancer, morphological changes seen in HPV lesions elsewhere are often seen. HPV DNA has been detected in 21.7% of the 2,468 bronchial carcinomas analysed to date and the same high risk types implicated in other squamous cell cancers have been identified. Clearly, more effort should be focused on assessing the role of HPV in bronchial carcinogenesis, by analysing the synergistic effects of carcinogenic agents (cigarette smoke, radiation, asbestos, etc) and HPV in different experimental settings.

Human papillomavirus lesions in association with cervical dysplasias and neoplasias.

A series of 620 cervical biopsy specimens (precancerous and malignant) was assessed morphologically with special reference to the concomitant appearance of human papillomavirus lesions. Tissue samples from 346 of these biopsy specimens were stained for human papillomavirus antigens using the immunoperoxidase-peroxidase-anti-peroxidase (PAP) technique. Papillomavirus lesions were found in 55.6% of the biopsy specimens associated with all degrees of epithelial atypia. The mean age of the women with papillomavirus (condylomatous) changes was significantly lower (P less than .0001) than that of women without these lesions, ie, those who had dysplasia/neoplasia without concomitant papillomavirus changes. Flat and inverted condylomas were most frequent between the ages 20 and 39 and were accompanied by more severe dysplasias than the papillomatous condylomas. In immunoperoxidase-PAP staining, 56% of the papillomavirus lesions were positive, the positivity being inversely related to the degree of epithelial atypia, and bearing some correlation with the condyloma type (papillary 100%, inverted 70%, and flat 52%). Although the results show a clear-cut association of human papillomavirus lesions with premalignant, and to a lesser extent with malignant squamous cell lesions of young sexually active women, thus suggesting a relationship between the virus and cancer, a careful follow-up study is needed to fully elucidate this relationship.

Expression of insulin-like growth factor II in female breast cancer as related to established prognostic factors and long-term prognosis.

The expression of insulin-like growth factor II (IGF-II) was analysed immunohistochemically in a series of 211 breast cancers with special reference to standard prognostic factors and patient survival. IGF-II was expressed both in the cancer cells and in stromal cells, in 84% and 50% of cases, respectively. IGF-II expression in cancer cells was related to a non-metastatic disease at diagnosis (p = 0.03), low tumour grade (p = 0.02), DNA diploidy (p = 0.02) and S-phase fraction under 7% (p = 0.001). IGF-II negativity was positively correlated to morphometric SD of nuclear area (p = 0.0003), nuclear perimetry (p = 0.002), SD of nuclear perimetry (p = 0.02), minimum nuclear diameter (p = 0.005) and maximum nuclear diameter (p = 0.003). The expression of IGF-II in stromal cells was related to low histological grade (p = 0.02), mitotic index under 10/mm2 (p = 0.01), mild nuclear pleomorphism (p = 0.03), DNA diploidy (p = 0.08), SD of nuclear area (p = 0.006), mean nuclear perimeter (p = 0.05), minimum nuclear diameter (p = 0.005) and maximum nuclear diameter (p = 0.007) in that the nuclear factor values were higher in tumours without stromal IGF-II expression. In univariate and multivariate survival analysis, immunohisto-chemically detected expression of IGF-II had no independent prognostic value over standard prognostic factors. Despite the fact that expression of IGF-II was inversely related to several histopathological features of malignancy, the clinical behaviour of breast cancer seemed to be independent of IGF-II expression.

Expression of p53 protein related to the presence of human papillomavirus (HPV) DNA in genital carcinomas and precancer lesions.

The E6 protein of the high-risk human papillomavirus (HPV) types 16 and 18 is capable of complexing with the wild-type p53 tumor suppressor gene product, leading to loss of the normal p53 function as an anti-oncogene, whereas the low-risk HPV types 6 and 11 lack this binding property. The malignant potential of HPV 16 and 18 has been ascribed to this complexing of E6 with p53, which regularly leads to undetectable expression of the latter in HPV-positive lesions. To assess the role of p53 in HPV-associated genital carcinogenesis, the expression of p53 protein was studied immunohistochemically in 22 genital carcinomas and precancer lesions; 8 vulvar carcinomas, 1 VIN (vulvar intraepithelial neoplasia), 5 cervical carcinomas and 8 CIN (cervical intraepithelial neoplasia) using monoclonal antibody PAb 1801. Presence of HPV was demonstrated by PCR using HPV consensus primers, and amplified HPV-DNA was digested with the restriction enzymes giving distinct patterns for various HPV-types in gel electrophoresis. HPV-typing was confirmed by in situ hybridization with biotinylated DNA probes. Altogether, 17 of the 22 specimens (77%) showed p53 expression: 67% of the precancer lesions and 83% of carcinomas. Expression was more frequent (89%) in the vulvar than (70%) in cervical lesions. Using PCR,HPV DNA was detected in 19/22(86%) of the samples. The following HPV types were identified: HPV 6 (2 samples), HPV 11 (3 cases), HPV 16 (5 cases), HPV 33 (3 cases), and 6 contained unidentified HPV types. All HPV DNA-negative specimens showed p53 expression. Of the 19 HPV DNA-positive lesions, 5 were p53-negative, three of these being HPV 16 positive CIN lesions. The remaining two HPV 16 lesions were invasive carcinomas with a weak p53 expression. HPV 6 and 11-positive lesions showed a weak p53 expression more frequently than HPV-negative cases and HPV 33 lesions. The results indicate that p53 expression is detectable, but it is less frequent and less intense in HPV DNA-positive genital precancer lesions and carcinomas (particularly those with HPV 16 DNA) as compared with HPV DNA-negative lesions.

Immunohistochemistry, in situ hybridization and polymerase chain reaction (PCR) in detecting c-myc expression in human malignancies.

The assessment of oncogene expression at cellular level is important in understanding the role of those genes in carcinogenesis. Using in situ hybridization and immunohistochemistry, the expression of oncogenes can be visualized in topographic relation to tissue morphology. In the present study, c-myc overexpression was studied in ten carcinomas of different origin (6 mammary adenocarcinomas, 2 vulvar and 2 bronchial squamous cell carcinomas) by in situ hybridization (ISH) with 35S-labeled RNA probes and by immunohistochemistry (IHC). DNA amplification and transcription of c-myc oncogene were also studied with polymerase chain reaction (PCR) using beta-globin as an intrinsic standard for DNA amplification. The effect of formalin fixation of c-myc expression was simultaneously studied. Half of the tumours (5/10) demonstrated c-myc mRNA overexpression by ISH performed on frozen sections and two of the samples were shown to over-express c-myc protein by IHC. Only two samples fixed in formalin showed positive signals for c-myc mRNA. None of the biopsies showed DNA amplifications either with ISH or PCR. The present results suggest that ISH with RNA probes is a useful method for detecting the transcription of activated oncogenes in malignant tissues, especially when applied on frozen sections. The results also indicate that in some cases, c-myc gene may be adequately transcribed to mRNA but the latter is not translated to the appropriate oncoprotein.

In situ DNA hybridization analysis of human papillomavirus (HPV) sequences in benign oral mucosal lesions.

A series of 144 surgically treated benign oral mucosal lesions were analysed using an in situ DNA hybridization technique with 35S-labeled human papillomavirus (HPV) DNA probes to demonstrate the DNA of HPV types 6, 11, 13, and 16, in routinely processed, paraffin-embedded biopsy specimens. These lesions and an additional 62 benign oral mucosal biopsy specimens (total, 206 specimens) were also assessed by the indirect immunoperoxidase (IP-PAP) technique to detect the expression of HPV structural proteins (viral antigens). A total of 54/206 (26.2%) lesions were observed to express HPV antigens, being found in 45/92 (48.9%) of the squamous cell papillomas/condylomas, in 1/54 fibrous hyperplasias, in 1/8 true fibromas, and in 7/8 (87.5%) of the focal epithelial hyperplasia (FEH) lesions. Of the HPV DNA-positive lesions, 15/45 (33.3%) expressed HPV antigens, the expression not being related to any particular HPV type. HPV DNA sequences were found in 45/144 (31.3%) of the lesions. HPV DNA was present with the highest frequency in FEH (83.3%), followed by the papilloma/condyloma group (33.8%), papillary hyperplasia (28.6%), fibrous hyperplasia (24.4%), and true fibromas (14.3%). The most frequent HPV type was HPV 11, representing 37.8% of the DNA-positive lesions. HPV 13 DNA, previously regarded as specific to FEH, was disclosed as a single HPV type in seven cases, and as a double infection by HPV 11 and 13 in an additional three cases, including all five morphologically distinct entities. Noteworthy is the discovery of the high-risk HPV type 16 DNA in 17.8% of the DNA-positive lesions, four papilloma/condyloma lesions, three fibrous hyperplasias, and one FEH.(ABSTRACT TRUNCATED AT 250 WORDS)

Value of morphological criteria in diagnosing cervical HPV lesions confirmed by in situ hybridization and hybrid capture assay.

The present study evaluated the value of morphological criteria (binucleation, multinucleation, koilocytosis, spindle koilocytes, abnormal mitosis and dyskeratosis) in the diagnosis of cervical human papillomavirus (HPV) lesions confirmed by in situ hybridization (ISH) and hybrid capture (HC) assay. Colposcopic punch biopsies from a series of 138 women with abnormal Pap smears were examined on light microscopy and in situ hybridization (DAKO widespectrum cocktail probe) for HPV-induced morphological changes and HPV DNA, respectively. Cervical swabs were analyzed for HPV DNA of the oncogenic types using Hybrid Capture. CIN 2 and CIN 3 were found in 44 biopsies, CIN 1 in 62, and no evidence of HPV in 32 cases. HPV was detected by ISH in 51/138 (37%) cases and by HC in 66/138 (48%) lesions. With both tests, HPV DNA detection increased parallel with lesion severity, up to 70% and 59% in CIN 2/3 by HC and ISH, respectively OR 4.6 (1.7-12.1) and 10.1 (3.0-33.8). Among the histological criteria, multinucleation, binucleation and abnormal mitoses were significantly associated with HPV DNA detection. Multinucleation proved to be the strongest predictor of HPV DNA-positivity. Binucleation, abnormal mitosis, koilocytosis and spindle koilocytes were also reliable criteria of HPV lesions. Minor nuclear atypia, and "mild koilocytosis" were of no value in making this diagnosis.

An osteogenic sarcoma of the thyroid gland (report of a case and survey of the literature).

An additional case, counted to be the twenty third, of a primary osteosarcoma of the thyroid gland in a 62-year-old woman is described. The literature on the subject is surveyed, and the possible histogenesis of this unusual neoplasm is discussed.