A new mode of treatment of human papilloma virus associated anogenital lesions using a nonsteroid estrogen analogue.

Human papilloma virus (HPV)-induced condylomata acuminata in both sexes, as well as cervical dysplasias were treated locally with an ointment or a cream containing 25 mg clomiphene citrate per gram for a fortnight. The DNA of HPV 6, 11, HPV 16, 18, and HPV 31, 35, 51 was monitored pre- and post-therapeutically using an in situ hybridization assay. The therapeutic response to the drug was demonstrated by the disappearance of condylomata acuminata (100%), HPV DNA in cervical dysplasia (up to 80%), and penile genital warts (75%). Under this therapeutic regimen 78% of all cases treated became negative for the probed HPV. The pharmacological action of the drug was diminished in HPV 16, 18 positive lesions. Clomiphene is shown to be an effective anti-HPV drug.

Effects of hypolipidaemics cetaben and clofibrate on mitochondrial and peroxisomal enzymes of rat liver.

Clofibrate or cetaben was administered to male rats for 10 days. Peroxisomal and mitochondrial enzymes were assayed in liver subcellular fractions. Clofibrate affected the specific activities of both mitochondrial enzymes (glycerol-3-phosphate dehydrogenase and nicotinamide-linked isocitrate dehydrogenase) and peroxisomal enzymes (fatty acyl-CoA oxidase, glycerone phosphate acyltransferase, urate oxidase, and D-amino-acid oxidase). In contrast, cetaben raised only the peroxisomal enzymes, acyl-CoA oxidase, glycerone-phosphate acyltransferase, D-amino-acid oxidase, catalase, and urate oxidase. Thus, the hypolipidaemic activity of these drugs may be exclusively related to stimulated peroxisomal functioning, while mitochondria play only a minor role.

[Synergistic cytotoxic effects of chemotherapy in colon tumor cells by simultaneous inhibition of de novo and salvage energy metabolism pathways]. / Synergistische zytotoxische Effekte einer Chemotherapie an Kolontumorzellen durch gleichzeitige Hemmung von de novo und salvage Stoffwechselwegen.

The success of chemotherapy of colon tumours is currently limited. We have therefore used the human colon tumour cell line HT-29 to evaluate the cytotoxic effects of various drug combinations. Trimidox (3,4,5-trihydroxybenzamidoxime), a recently patented inhibitor of ribonucleotide reductase was combined with cytosinearabinoside (Ara-C) or 2',2'-difluorodeoxycytidine (DFDC) in order to inhibit both pyrimidine de novo and salvage pathways. Synergistic cytotoxic effects were observed. When HT-29 cells were sequentially treated with trimidox (20 microM for 24 h) and Ara-C (2 microM for 2 h), colony numbers decreased to 71% of the value calculated for additive cytotoxicity. When cells were simultaneously treated with trimidox (10 microM and 15 microM) and DFDC (0.2 nM), synergistic inhibition of colony formation was likewise noted (colony numbers decreased to values as low as 73% or 71% of the values calculated for additive cytotoxicity). On the other hand, we combined tiazofurin, an inhibitor of the guanylate de novo pathway, with allopurinol, which inhibits the guanylate salvage pathway by increasing intracellular hypoxanthine concentrations, leading to inhibition of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT). Synergistic cytotoxic effects were observed under these conditions too. When cells were treated with 10 microM tiazofurin and 400 microM or 800 microM allopurinol the number of colonies decreased to 69% and 27%, respectively, of the values calculated for additive effects. Our data suggest these drug combinations to be promising options in the treatment of human colon cancer.

Clofibrate elevates enzyme activities of the tricarboxylic acid cycle in rat liver.

Activities of the tricarboxylic acid cycle enzymes were measured in subcellular fractions of liver from rats that had been fed clofibrate for 3 weeks. Large changes in these activities per gram tissue were found in the large particle fraction, which also showed an increase in total protein concentration of 76% under clofibrate treatment. The three regulatory enzymes of the cycle, namely citrate synthase, NAD(+)-linked isocitrate dehydrogenase, and 2-oxoglutarate dehydrogenase were significantly enhanced by 24% (P < 0.02), 54% (P < 0.02), and 153% (P < 0.005), respectively. Fumarase and malate dehydrogenase rose by 71% (P < 0.005) and 95% (P < 0.02), whereas succinate dehydrogenase remained unchanged. Enhancement of the citrate synthase, NAD-isocitrate dehydrogenase, and 2-oxoglutarate dehydrogenase may play a role in decreasing intracellular availability of acetyl-CoA for lipid metabolism.

Beta-endorphin, steroids, and prolactin. Immunoassay in breast cysts and blood.

Circulating levels and cyst fluid concentrations of beta-endorphin (beta-EP), estradiol, progesterone, and prolactin were measured by radioimmunoassays in 10 premenopausal women with gross breast cysts. In addition, aspirates and frozen tissue sections from cystic lesions were investigated immunocytochemically for a possible beta-EP production. Epithelia of dilated ducts, smaller cysts, and hyperplastic lesions without atypia showed a strong positive reaction in the cytoplasm. In apocrine metaplasia, this staining was concentrated in the apical region. The staining intensity in atypical hyperplasia was diminished. Occasionally, normal duct and lobular epithelia exhibited positive beta-EP immunostaining. Levels of beta-EP, estradiol, and progesterone in the cyst fluid were significantly higher than in blood, but no significant differences were observed for prolactin. The ratios of progesterone to estradiol, estradiol to prolactin, and progesterone to prolactin in the cyst fluid were considerably higher than in blood. This suggests that beta-EP and steroid hormones are secreted from the lining epithelia into the breast cysts.

Endovaginal Doppler flow measurements of the ovarian artery in patients with a normal menstrual cycle and with polycystic ovary syndrome during in vitro fertilization.

Women with a normal menstrual cycle (n = 21, controls), polycystic ovary syndrome (n = 10) and hypogonadotropic amenorrhea (n = 3) were stimulated with clomiphen-citrate (4th day to 8th day of the cycle) and with human menopausal gonadotropin (8th day to 11th day). The vascular impedance of the ovary carrying the dominant follicles was monitored by endovaginal pulsed Doppler flow measurement. Simultaneously, serum levels of LH, E2 and 17-OHP were assayed. Contrary to controls, women with polycystic ovary syndrome or hypogonadotropic amenorrhea showed decreased hormone levels and no lowering of the vascular impedance. In controls, the lower pulsatility index is caused by neovascularization around the dominant follicle and by E2-induced vasodilatation in the ovarian artery.

[Diagnosis of cephalothoracopagus--a case report]. / Diagnose eines Kephalothorakopagus--Eine Kasuistik.

Cephalothoracopagus twins were diagnosed at 22 weeks' gestation, hence this case was diagnosed early enough antenatally to allow vaginal pregnancy termination. This case underlines the need for exact ultrasound screening during early pregnancy.

Screening for Epstein-Barr and human cytomegalovirus in normal and abnormal cervical smears by fluorescent in situ cytohybridization.

Using the routine Papanicolaou test for grouping, 35 normal and 148 abnormal cervical smears were screened by fluorescent in situ cytohybridization for Epstein-Barr (EBV) and human cytomegalovirus (HCMV). Viral presence was detected in 55% of all hybridized smears while routine Papanicolaou tests exposed no morphological evidence for HCMV or EBV. Thus in situ cytohybridization is necessary and suitable for direct identification of EBV and HCMV. The prevalence of viral infection for women with normal cervical smears was 29% for HCMV and 34% for EBV. Even smears of three of the four virgins were positive for HCMV and two had a mixed infection with EBV. The infection rate with at least one of these two herpesviruses was 51% in the control group, 56% in smears with reactive and reparative cell changes, 54% in mild (slight) dysplasias and 56% in moderate or severe dysplasias. Thus infection with HCMV and EBV did not differ significantly in controls and prevention groups. Both viruses were most frequently found in the age group 20 to 29 years. These data suggest that HCMV and EBV are not involved in the etiology of intraepithelial neoplasias and carcinomas of the uterine cervix.

Enhancement of NAD-linked isocitrate dehydrogenase activity in rat liver by clofibrate feeding.

After feeding rats for 3 weeks with clofibrate specific activities of the liver mitochondrial enzymes glycerol-3-phosphate dehydrogenase (G3PDH) and NAD-linked isocitrate dehydrogenase (NAD-ICDH) were found to be increased in a large particle fraction 6-fold and 1.6-fold, respectively, whereas the activity of NADP-linked isocitrate dehydrogenase (NADP-ICDH) remained unchanged. Possibly these effects contribute to the hypolipidemic action of clofibrate.

Breast cyst fluid concentrations of beta-endorphin, steroids and gonadotrophins in premenopausal women with gross cystic disease.

Breast cyst fluid (BCF) and plasma levels of beta-endorphin (beta-EP), oestradiol (E2), progesterone (P), luteinizing hormone (LH), follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), prolactin (PRL) and cortisol were assayed radiochemically in a group of 10 premenopausal women aged 45-50 years suffering from gross cystic breast disease. The concentration of beta-EP (fmol/ml) in BCF (17.6 +/- 4.6 S.E.) was over four times higher than that in plasma (4.2 +/- 0.5 S.E.). The level of E2 was 41 times higher (1738.2 +/- 350.5 S.E. pg/ml) and that of P was 47 times higher (65.47 +/- 8.25 S.E. ng/ml) in BCF than in plasma. The significantly increased values of beta-EP, E2 and P in BCF and the identification of beta-EP in the cyst-lining epithelium indicated that local synthesis occurs. Growth factor-like properties of beta-EP and E2 are responsible for the propagation of cystic changes. The autonomous formation and activity of beta-EP, E2 and P in cystic formations were not correlated with LH, FSH, TSH and cortisol levels, which were significantly higher in plasma than in BCF. The concentration of PRL in BCF was not significantly higher than that in plasma. Moreover, the plasma concentrations of testosterone, androstenedione, thyroxine, triiodothyronine, thyroxine-binding globulin and sex-hormone-binding globulin were within the normal ranges. In this study we demonstrated synergism between beta-EP and steroid hormones which encourages the development of gross cystic disease in premenopausal women.

Response to thyroxine of lamellar bodies, peroxisomes and peroxisomal enzymes in the adult rat lung.

Adult male rats were fed a standard diet containing 25 mg/kg L-thyroxine for 2 weeks. The hyperthyreotic condition of the animals was checked by monitoring the metabolic rates and liver glycerol-3-phosphate dehydrogenase. In the postnuclear fraction of the lung the activity of fatty acyl-CoA oxidase, the enzyme responsible for the rate limiting first step of peroxisomal fatty acid beta-oxidation, showed a twofold increase. Catalase, the marker enzyme of peroxisomes, showed a similar increase. Electron microscopic examination of alveolar type II cells did not reveal changes in the number and distribution frequency of peroxisomes and lamellar bodies. Similarly the content of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoyl phosphatidylcholine), the main constituent of alveolar surfactant, was not altered significantly by thyroxine feeding. On the other hand the volume density of the peroxisomal compartment was found to be doubled according to the measured increase of catalase and acyl-CoA oxidase. Our data suggest that the induction of peroxisomal matrix enzymes, such as catalase and fatty acyl-CoA oxidase, does not influence the surfactant content.

Papanicolaou test and enzyme-linked in-situ hybridization. A combined diagnostic system for papilloma virus infections with high prognostic value.

A combined diagnostic system for human papilloma virus (HPV) infections comprising the Papanicolaou test and in-situ hybridization assay was evaluated. Cervical smears from 259 women obtained with a "Cytobrush" were screened. Human papilloma virus genotypes 6/11, 16/18, 31/35/51 were detected by biotin in-situ hybridization in conjunction with a streptavidin-alkaline phosphatase detection complex. The diagnostic sensitivity of this assay was tested by human papilloma virus-DNA-positive human cervical carcinoma cell lines. According to the cytological (Bethesda system) and colposcopical criteria a random control group (n = 80) and prevention (n = 179) were chosen. Compared with Papanicolaou tests the frequency of human papilloma virus-DNA-positive cervices rose with the severity of cell abnormalities. The detection rate of human papilloma viruses-16/18 and human papilloma viruses-31/35/51 and of concomitant infections with human papilloma viruses-6/11 and human papilloma viruses-16/18 and/or human papilloma viruses-31/35/51 increased with the severity of cell dysplasia, whereas the rate of human papilloma virus-6/11 DNAs decreased. The incidence of oncogenic human papilloma virus types 16/18 and 31/35/51 rose with the age of the patients. A follow-up study by Papanicolaou tests of patients with mild (slight) and moderate dysplasias six months after human papilloma virus-DNA-hybridization indicates that human papilloma virus-16/18 DNA-positive lesions are more likely to persist or to progress than human papilloma virus-6/11 DNA-positive cell changes. Human papilloma virus-31/35/51 DNA-positive cell smears exhibited persistent behaviour. Our findings demonstrate that the Papanicolaou test combined with in-situ hybridization is suitable for early diagnosis and prevention of intraepithelial neoplasias and carcinomas of the uterine cervix.

Reliability of in-situ hybridization of smears and biopsies for papilloma virus genotyping of the uterine cervix.

Two sampling methods, biopsy and cell smear, were investigated for their reliability in the biotin in-situ hybridization test for human papilloma virus deoxyribonucleic acid (HPV-DNA)-6/11 and 16/18. Cervical smears and biopsies were obtained simultaneously from 81 women with cervical lesions. The sensitivity of the in-situ hybridization was tested on human cervical carcinoma cell lines. As a reference method, HPV-DNA was probed in biopsies using a Southern blot with 32P-labelled DNA probes. This reference method detected HPV-DNA in 45% of the reactive and reparative cell changes, in 75.9% of intraepithelial neoplasias, and in 83.3% of squamous cell carcinomas of the cervix. Of the examined cervices 61.7% were HPV-DNA-positive. As tested by in-situ hybridization, 56.8% of the biopsies were HPV-DNA-positive. Three biopsies were HPV-DNA-negative by the in-situ hybridization but positive by the Southern blot. One biopsy was HPV-DNA-6/11 positive by the in-situ hybridization but negative in the Southern blot. As tested by in-situ hybridization, 55.6% of the cervical smears were HPV-DNA-positive. Five smears were HPV-DNA-positive by the in-situ hybridization but negative by the reference method, thus demonstrating the dependence of the HPV positive rate on the sampling method. Four cell smears were negative by the in-situ hybridization but positive by the reference method, which shows that the biotin in-situ hybridization is less sensitive. The reference method confirmed the HPV-DNA-positivity of in-situ hybridization for 97.8% of biopsies, and for 91.8% of cell smears.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunochemical and ultrastructural studies of an ovarian strumal carcinoid.

An ovarian strumal carcinoid which synthesized peptide hormones, but did not induce the carcinoid syndrome, was analysed histochemically, immunohistochemically and ultrastructurally. Dot-immunobinding assays were performed in order to determine the endocrine gene expression. The amylase resistant colloid was found to be PAS-positive in the follicular portions of the tumour. Carcinoid cells showed Grimelius positive argyophilic granules in the subnuclear position. The Fontana-Masson argentaffin reaction was negative. Immunohistochemistry for adrenocorticotropic hormone (ACTH) revealed strong reactivity in the follicular areas of the carcinoid. The immunoreactivity for somatotropic release inhibiting factor (SRIF) was found positive in the trabecular portion of the carcinoid tumour, thyroglobulin in the follicles. Neuron-specific enolase, protein S-100 A/B, synaptophysin and chromogranin A evoked weak cytoplasmic immunostaining of the tumor cells. Dot-immunobinding assays substantiated these immunohistochemical results, except for the thermolabile protein S-100 A/B. Electron microscopy of tumor cells showed numerous electron-dense cytoplasmic granules, 250 to 350 nm in diameter, both in follicular and trabecular areas of the tumor. Plasma levels of tumor-associated ACTH, SRIF and thyroglobulin were measured by radioimmunoassay and were found to be within the normal range.

[The value of vaginal ultrasonography in early pregnancy]. / Die Bedeutung der Vaginosonographie in der Frühschwangerschaft.

Heart action was detected in our first group of 150 pregnant women during the early period of pregnancy (5th to 8th week post menstruationem) in 25% at the end of the 5th week of pregnancy, in 85% at the end of the 6th and, in 100% in the 7th week. In our second group of 59 women with tubal pregnancy, confirmed by laparoscopy, it was possible to detect the extrauterine pregnancy transabdominally in 25% and transvaginally in 94.7% via sonography (p = 0.03). Extrauterine pregnancy was easier detectable by vaginosonography due to the proximity of the organ and the better power of resolution, as a direct sign of extrauterine ring formation surrounded by a circular wall-like structure.

Biotin and phosphorus-isotopic labelled DNA/RNA probes for the detection of human papilloma virus sequences.

In this study, the diagnostic accuracy and practicability of different hybridization techniques for the detection of human papilloma virus (HPV) DNA were tested. Cervical cell scrapes (n = 67) were analysed for HPV-DNAs 6/11 and 16, in order to compare a commercially available in situ DNA hybridization-assay with the conventional Southern-blot analysis. The in situ DNA hybridization-assay gave a sensitivity of 81.5%, a specificity of 97.5% and a diagnostic efficiency of 91.0% for HPV-DNAs 6/11. Using the same assay, we observed a sensitivity of 100%, a specificity of 96.3% and a diagnostic efficiency of 97.0% for HPV-DNA 16. The practicability of dot-blot DNA hybridization technique was tested on 176 cervical cell scrapes, in order to determine the prevalence rate of HPV-genotypes 6/11, 16/18 and 31/33/35. In the random control group (n = 106), 1.9% of the cases were HPV-DNA positive. In the cancer prevention group (n = 70), patients with reactive and reparative cell changes showed a HPV-DNA positivity of 55.0%, with mild (slight) dysplasia/CIN 1 of 73.7%, and with moderate to severe dysplasia/CIN 2 to CIN 3, including the carcinoma in situ/CIN 3 of 80.0%. Patients with squamous cell carcinoma of the cervix uteri gave HPV-DNA positive results in 96.2% of the cases. The suitability of in situ DNA hybridization for morphological studies was tested on tissue biopsies (n = 68). The HPV-DNAs 6/11 were found predominantly to 72.7% of the examined condylomas. The HPV-DNA positive cervices increased with the severity of the cytological dysplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Early detection of ectopic pregnancy by transvaginal ultrasound.

Transvaginal (TVU; 19 patients) and transabdominal ultrasound (TAU; 24 patients) were compared in their ability to visualize an ectopic pregnancy. The direct visualization of ectopic pregnancy (EUP) was 25% with TAU and 94.7% with TVU. All women had a proven EUP in laparoscopy. TAU approach showed a significant inferiority (P less than 0.02) and TVU a superiority (P less than 0.08) in comparison to clinical examination. 83% of 43 women had spotting. Nonspecific signs for EUP like fluid in the pouch of Douglas was seen in 20.9%. Thin endometrium was seen in 6.9%, in 37.2% the thickness of endometrium was under 10 mm, in 55.8% more than 10 mm. An intrauterine pseudogestation sac was detected in 6.9%, corpus luteum formation in 37.2%.

[The significance of human papillomavirus infection in cervix cancer and its preliminary stages (including other HPV-induced genital lesions)]. / Die Bedeutung humaner Papillomavirus-Infektionen beim Zervixkarzinom und seiner Vorstufen (einschliesslich anderer HPV-bedingter genitaler Läsionen).

Southern blot analysis, DNA dot blot technique and in situ hybridization were used to examine cells and tissues from lesions of the female genital tracts for infections of human papillomaviruses (HPV) of the types 6/11, 16/18 as well as 31/33 and 35. As the degree of dysplasia increased the proportion of HPV-positive results increased significantly, especially for the HPV types 16/18; there was a decrease in the HPV DNA 6/11-positive cases. In the first follow-up examinations of cervical smears of the Pap. IIID type after 6 months there was a persistence of 84.2%. Of these, 57.8% were HPV 16/18-positive, and 18.2% could be classified as belonging to Pap. group IV. The importance of HPV typing lies in the definitions of the pathogens, risk groups, therapeutic measures and their controls.