One hundred and fifty-eight histologically verified mammary carcinomas with known mammographic doubling time (DT) were studied with special emphasis on a morphologic classification proposed by Linell et al. [8, 12, 14, 15]. The hypothesis that Linell classification of ductal carcinomas into comedo, tubuloductal and tubular carcinomas is easy to perform with small inter-observer variations, was not fully confirmed. The Linell classification was found to correlate well with conventional WHO malignancy grading, S-phase fraction and DNA-ploidy. The Linell classification also correlated to surgical stage, lymph node status and DT, but not at all to tumour size. Using distant disease-free survival as an endpoint, the Linell classification gave prognostic information comparable to conventional histologic grading, seeming to be a simple, cheap and reliable method well worth trying on a larger scale.

More than a decade ago an association between acute intermittent porphyria (AIP) and hepatocellular carcinoma (HCC) was reported, but still the cause of the increased prevalence is unknown. Paraffin sections of formalin-fixed HCC from 17 AIP patients were reexamined and also screened for relevant mutations using several methods. The tumor diagnosis was verified, and in several cases precirrhosis and cirrhosis were also found. The clinically founded AIP diagnosis was verified at the gene level in most cases, demonstrating the Norrland type of mutation, i.e., G(593)-to-A substitution in codon 198 of the porphobilinogen deaminase (PBGD) gene. The second allele was neither mutated nor missing, contradicting the possibility that the PBGD gene might function as a tumor suppressor gene. Subsequent sequencing showed that cases not cleaved by the restriction enzyme NheI lacked the specific Norrland mutation. In recent years, selective mutations at codons 249 and 166 of the p53 gene have been described in HCC associated with aflatoxin and hepatitis B virus. In our area, with low exposure to those agents, no mutations in codon 249 were found, and mutation in codon 166 was excluded in all tumors except one; no traces of hepatitis B DNA were observed. Nor did we find mutations in H-ras 12 or 61. Intrinsic aberrations in AIP, including reduced heme synthesis and endogenous oxidative damage to DNA, may incite carcinogenic mutations elsewhere in the genome of liver cells. The increased cell proliferation coupled to precirrhosis and cirrhosis perhaps represents promotion in the initiation-promotion sequence of hepatocarcinogenesis.

Background. The authors examined prognostic factors in 158 cases of breast carcinoma with known mammographic tumor volume doubling times (DT).Methods. The tumors were retrospectively reexamined histologically and flow cytometric analysis of DNA ploidy and S-phase fraction (SPF) was performed on archival paraffin-embedded material in each case. Life tables and Cox multivariate analyses were used for statistical evaluation of prognostic factors.Results. In univariate analysis of survival data, clinical and pathologic stage, histologic grade, the presence of axillary lymph node metastases, and SPF were significant prognostic predictors, but mammographic DT and DNA ploidy were not. SPF also contributed prognostic information in the subgroup of carcinoma cases detected by screening. In a Cox multivariate analysis, SPF, the presence of axillary lymph node metastases, and Stage II-III disease (as opposed to Stage I disease) were independent significant predictors of survival. In univariate analyses of distant disease-free survival, clinical and pathologic stage, tumor size, histologic grade, the presence of involved axillary nodes, DT, and SPF all were significant prognostic factors.Conclusions. SPF, stage, and lymph node status were important prognostic factors in this patient material with predominantly small and node-negative breast carcinomas, whereas DNA ploidy and mammographic DT provided less prognostic information. The prognosis of carcinoma detected during screening did not differ significantly from that of breast carcinoma discovered otherwise in this selected patient group.

Background. In a retrospective study, correlations among mammographic doubling times (DT), clinicopathologic prognostic factors, and cytometric predictors were examined.Methods. One hundred fifty-eight patients with the possibility to calculate mammographic tumor DT were selected and the tumors were histologically reexamined and flow cytometric analysis for ploidy and S-phase fraction (SPF) was performed.Results. The tumors were Stage I in 68%, and 45% were detected by mammographic screening. DT ranged from 0.6 months to an indefinite time (median, 9.0 months). Short DT was significantly correlated to large tumor size (P = 0.01) and advanced pathologic tumor stage (P = 0.016), but there was no correlation between DT and histologic grade. Ploidy analysis indicated that there were 57% aneuploid and 7% tetraploid tumors. There was a significant overrepresentation of euploid tumors among tumors smaller than 10 mm (P = 0.02). Ploidy was correlated to histologic grade (P < 0.001) and DT (P = 0.009). SPF was calculated in 122 cases. SPF correlated significantly with pathologic stage (P = 0.002), tumor size (P = 0.037), histologic grade (P = 0.001), the presence of axillary lymph node metastases (P = 0.046), DT (P = 0.02), and DNA ploidy (P < 0.001). Compared with interval carcinoma, screening-detected carcinoma showed favorable characteristics concerning size, stage, DT, ploidy, and SPF but not regarding histologic grade and axillary lymph node metastases.Conclusions. DT shows great variations. Factors related to tumor biology (i.e., DT, DNA ploidy, and SPF) are strongly correlated with one another, but they have no correlation with axillary lymph node metastases. Cancer detected by screening is discovered at an early stage and shows favorable characteristics concerning DT, ploidy, and SPF.

Biopsy samples from 13 Kenyan patients with squamous cell carcinoma of the cervix were analysed for the presence of type specific HPV DNA by polymerase chain reaction (PCR). HPV 16 was confirmed in 11 (85%) and HPV 18 in 9 (69%) samples. HPV 6 DNA was detectable in only 3 (23%) samples and no HPV 33 was found. Infection with either HpV16 or 18 was seen in 12 (92%) and infection with both in 8 (62%) cases. The prevalence of double infection found is higher than in previous reports. The significance of this and possible effects of parity on cervical neoplastic changes are discussed.

Human papillomavirus (HPV) type-specific sequences required for polymerase chain reaction (PCR) mediated amplification of HPV DNA sequences are presented. One primer pair within the E1 open reading frame (ORF) was shared by HPV 6, HPV 11, HPV 16, and HPV 31, whereas the other primer pair within the E1 ORF was specific for HPV 16. Eight primer pairs from the E6 and E7 ORFs specifically detected HPV 6, HPV 16, HPV 18, and HPV 33 sequences. This system has been used for detection of HPV DNA in biopsies, cytological smears and sections of formalin-fixed tissues.

The action of LH is mediated through specific plasma membrane receptors that are both up- and down-regulated in the ovary during the reproductive cycle. Using immature rats treated with PMSG and hCG as a model system, we have studied the regulation and distribution of LH receptor mRNA in different cell types during follicle development, ovulation, and luteinization by Northern blot and in situ hybridization. In untreated rats, LH receptor mRNA was below the detection level in granulosa cells, cumulus cells, and oocytes, while low levels of LH receptor mRNA were found in the thecal cells. After stimulation with PMSG, expression of LH receptor mRNA was enhanced in the thecal-interstitial cells, while a more dramatic increase in receptor mRNA abundance took place in granulosa cells of large tertiary follicles. In these follicles, the abundance of LH receptor mRNA varied among different subpopulations of granulosa cells, with mural granulosa cells close to the basement membrane exhibiting higher levels than granulosa cells located closer to the antrum, and cumulus cells and the oocyte lacking detectable hybridization signal. The uneven expression of LH receptor mRNA endows different ovarian cells with varying hormonal responsiveness. After an ovulatory dose of hCG, LH receptor mRNA levels were dramatically decreased, particularly in the granulosa cells of preovulatory follicles, to reach the lowest levels just before ovulation. During the transformation of ovulated follicles into corpora lutea, the expression of LH receptor message was again increased. Our results reveal that the previously documented regulation of the LH receptor-binding activity during ovarian development correlates with expression of the LH receptor transcripts, suggesting that the LH receptor gene is regulated in a complex manner during the periovulatory period to achieve cell-specific expression together with gonadotropin induction and suppression of receptor gene activity.

Using Southern blot or in situ hybridization, several studies have recently reported an association between HPV 18 and adenocarcinoma of the uterine cervix. In this study PCR was used to analyse the presence of HPV 16 and 18 in paraffin embedded biopsies of cervical adenocarcinoma in Northern Sweden. HPV DNA was confirmed in 11 (42%) of 26 cases. Seven (27%) were positive for HPV 18 and 4 (15%) for HPV 16. Nine of 13 premenopausal cases were HPV positive compared to only 2 of 13 postmenopausal cases (p less than 0.015). There was also a tendency towards previous negative cytology in HPV 18 positive premenopausal patients. The significance of these findings in relation to epidemiology of cervical adenocarcinoma is discussed.

Various methods presently available for the diagnosis of genital Human Papilloma Virus (HPV) were compared regarding their sensitivity in women referred for specific diagnosis and treatment because of atypical Pap smears or clinically suspected neoplasia.Colposcopic examination was performed in all cases. In addition to taking a second Pap smear, cell suspensions were made from 105 women and analysed by the Filter In Situ Hybridization (FISH) technique and tested for HPV 6+11 and HPV 16+18+31. The FISH technique was also used for the possible detection of HPV‐DNA in a reference material comprising 119 apparently healthy women with normal Pap smears.Colposcopically directed cervical biopsies in altogether 196 specimens were obtained from 155 women for histopathological examination and also for the detection of HPV‐DNA by the Southern blot hybridization technique. These specimens were tested for the presence of HPV 6, 11, 16, 18, 31 and 33.Three per cent of the women with and 34% without cytological signs of HPV in Pap smears had cervical intraepithelial neoplasia (CIN) 111 according to histopathology. CIN III was present in 35% of biopsies with and 59% of biopsies without histological signs of HPV in the biopsies.Altogether 46% of the women were HPV‐DNA positive. Of the women analysed by Southern blot, 39% were HPV‐DNA positive. Of the samples analysed by FISH, 27% with atypical cells were HPV‐DNA positive, compared with 11% of the samples from reference women with normal cytology.Of the women positive for HPV 16/18, colposcopy revealed HPV signs and or dysplasia in 95%.The presence of HPV in precancerous epithelium is not easily confirmed, either by cytology, colposcopy, or histopathology.By using Southern blot technique, it can be established that HPV‐DNA (67%) is frequently present in CIN III lesions. The FISH technique appeared to be as sensitive as Southern blot concerning the possibility of detecting HPV‐DNA in benign tissue and neoplasia of mild to moderate degree.

DNA analysis with static and flow cytometry was performed on archival smears and tissue sections in 99 patients with T2 breast cancer (Stage II). Tumour size, histologic grade and axillary node metastases were significant prognostic predictors. Static cytometry revealed 63% aneuploid tumours, and ploidy was significantly correlated to histologic grade and survival. DNA measurements obtained by static and flow cytometry were strongly correlated. According to flow cytometry 53% of the tumours were aneuploid. Flow cytometric DNA analysis correlated to histologic grade and survival and gave prognostic information among the lymph-node negative patients. Ploidy seems to be a significant, although not an independent prognostic indicator for T2 breast cancer.

Malignant transformation is reported in less than 2% of benign cystic teratomas. Although all the elements can undergo this transformation, it is most often seen in squamous epithelium. The malignancy of neural elements is probably the least common event, with only one case previously reported. A case of glioblastoma multiform in a benign cystic teratoma is presented. Its neural derivation was supported by an immunohistochemical staining specific for Glial Fibrillary Acidic Protein (GFAP). Despite very conservative surgery without adjuvant therapy, the patient remains alive and symptom free more than 3 years later.

The prognostic value of DNA analysis was studied retrospectively in 91 patients with locally advanced breast cancer (T3, T4) and a follow-up time of 3-7 years. Tumor cell DNA analysis was performed by static cytometry on aspiration biopsy specimens in 42 cases and on tissue sections in 49 cases. The tumors were classified as euploid or aneuploid. Sixty-four percent of the tumors were aneuploid. DNA pattern correlated significantly to histologic grade and axillary perinodal growth and also to survival. DNA pattern, histologic grade and axillary node metastases correlated significantly to disease-free survival (DFS). In this series of patients with locally advanced breast cancer DNA determinations gave important prognostic information.

Although treatment of cultured granulosa cells with gonadotropins increases their fibrinolytic activity, the biochemical nature of this effect is unclear. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fibrin autography techniques to characterize the fibrinolytic components secreted by granulosa cells. The fibrinolytic activity of these cells results from the production of both a tissue-type plasminogen activator (t-PA) and a urokinase-like activator (u-PA). The cells also produce an inhibitor of fibrinolysis (antiactivator). FSH and LH stimulate t-PA activity and suppress antiactivator activity, while u-PA activity is not affected by the gonadotropins. The differential regulation of these molecules by the gonadotropins may be essential for ovulation.

Isolated ovaries from untreated, sexually mature rabbits were introduced into an in vitro perfusion system and perfused with a chemically defined medium containing albumin. The ovaries were perfused for up to 15 h (mean 11.5 h) and then processed for morphological investigation. Both at the light- and electron-microscopical levels, most of the ovaries exhibited a normal structure comparable with ovaries in situ. In two cases, however, marked accumulations of bacteria were found, although not inside the follicles. Since ovulation in the rabbit normally occurs between 9.5-13 h after mating or human chorionic gonadotrophin treatment, this model seems adequate for studies of ovulation in vitro. It is, however, important to study the ovaries microscopically after the perfusion to detect artifacts, e.g., bacterial infection, that may have influence on the process of ovulation.

Both ovaries of 31 rabbits were perfused with a chemically defined medium in vitro in a recirculation system. In one series of experiments, hCG (100 IU) was injected iv 5-6 h prior to anaesthesia and surgery. Approximately 1 h later the perfusion was started. One ovary was perfused as control while the other ovary was perfused with 5 micrograms/ml indomethacin or with indomethacin and 1 micrograms/ml PGF2 alpha. In another series of experiments the rabbits received no pretreatment prior to operation. Instead, bovine LH was added to the perfusion medium of both control and experimental ovaries. The experimental side also received either indomethacin or indomethacin and PGF2 alpha. Finally, the effect of PGF2 alpha in the absence of LH was compared to the control ovary receiving only LH. After injection of hCG in vivo, ovulations occurred in 4 of 5 control ovaries. Indomethacin completely blocked ovulation in 4 of the 5 ovaries treated, while PGF2 alpha restored ovulations in all the experimental ovaries. In the group of experiments where LH was added in vitro, ovulations were induced in all ovaries treated with varying LH doses. Furthermore, indomethacin blocked ovulation in 5 out of 7 ovaries, and PGF2 alpha restored ovulation in all ovaries. Fifty per cent of the ovaries treated only with PGF2 alpha (in the absence of LH) also ovulated. The pattern of steroid release did not differ between control ovaries, indomethacin treated ovaries, and indomethacin + PGF2 alpha treated ovaries. Ovaries treated in perfusion with PGF2 alpha alone had very low steroid levels compared to the ovaries treated with LH.(ABSTRACT TRUNCATED AT 250 WORDS)

Using a method of in vitro perfusion of the rabbit ovary with a chemically defined medium in a recirculation system, normal appearing follicular ruptures occurred following exposure of the ovaries to hCG in vivo (100 IU) or following addition of LH (0.25 microgram/ml of NIH B9) to the perfusate. The addition of oestradiol-17 beta (10 micrograms/ml) to the perfusate did not inhibit these follicular ruptures, although the follicular fluid oestradiol contents were increased more than 100-fold as compared to the control side not receiving the addition of oestradiol. These data suggest that the physiological decline of follicular oestrogen, normally observed in vivo prior to ovulation in the rabbit, is not required as part of the mechanism of ovulation and that normal appearing ovulations can occur even though follicular oestrogen levels are kept artificially elevated.