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The report describes atypical multinucleated giant cells adjacent to proliferated nerve fascicles in a circumscribed subepithelial area of the cervix uteri of a 44-year-old multipara. Ultrastructural examination revealed cytoplasmatic processes, basal lamina, intracytoplasmic microfibrils, bizarre nuclear shapes with pseudoinclusions and nuclear fragments connected by small chromatin bridges (nucleotesimals). Immunohistochemical examination showed positive staining for vimentin and S-100 protein. Quantitative topography exhibited an isotropic distribution of the giant cells in an anisotropic architecture of mononuclear cells. A Schwann cell origin of the atypical giant cells is postulated. Aetiopathogenetically the lesion is regarded to be due to a trauma during delivery followed by regenerative proliferation of nerve fascicles and degenerative alterations of proliferating Schwann cells. The knowledge of this lesion is considered important, because the atypical cells could be confounded with malignant neoplastic cells.

Culture of cervical tissue with radiolabelled precursors showed that, in pregnant ewes, net hyaluronate synthesis increased between days 120-140 but that the marked polydispersity of hyaluronate remained unchanged. In the less sulphated of two proteoglycan populations, the proportion of heparan sulphate increased and that of dermatan sulphate decreased, while the less polydisperse more highly sulphated species showed a significant increase in molecular weight with pregnancy.

The authors describe the detection of human papillomavirus (HPV) 16 DNA in paraffin-embedded, formalin-fixed tissues of cervical squamous intraepithelial lesions (SILs) by in situ hybridization after amplification by the polymerase chain reaction (PCR). Using conventional in situ hybridization and a biotin-labeled probe, variable numbers of superficial cells and none of the basal cells in the SILs showed detectable HPV 16 DNA. When the in situ assay was done after amplification, increased numbers of superficial cells had detectable HPV DNA, and the hybridization signal was much more intense. HPV DNA was also detected in basal and parabasal cells at the site of the lesion whereas not detectable in directly adjacent, normal squamous epithelium. Amplified HPV DNA was demonstrated in formalin-fixed SiHa cells using a biotin-labeled probe, demonstrating the ability to detect one copy of HPV 16 DNA. This technique should allow for direct visualization in cells of other DNA sequences of low copy number from achival specimens otherwise undetectable by conventional in situ hybridization analysis.

Adult T-cell leukemia-derived factor (ADF) is an autocrine interleukin-2 receptor-inducing factor produced by human T-lymphotropic virus-1 (HTLV-1)-transformed lymphocytes, which has a high structural homology with an endogenous dithiol reducing coenzyme, thioredoxin. Its localization was investigated immunohistochemically in the cervix, using normal tissue (27 samples) and squamous neoplastic tissue (three condylomas, 42 cervical intraepithelial neoplasia [CIN] samples, 34 invasive squamous cell carcinoma samples). The expression of human papillomavirus (HPV) DNA was also studied in serial sections of the same subjects. Normal squamous cells and glandular cells of the cervix were negative for ADF. However, intracytoplasmic and/or intranuclear ADF-positive cells were usually found in the intermediate and superficial layers of the neoplastic squamous epithelium of condylomas (three of three cases) and CIN (35/42 cases). HPV DNA was detected in all condylomas and in 27 of 42 CIN specimens. HPV DNA-positive cells were usually localized in the intermediate and superficial layers of the neoplastic squamous epithelium. These HPV DNA-positive cells were also positive for ADF. Invasive squamous cell carcinoma was also positive for ADF (24/34 cases) and HPV DNA (11/34 cases). The coexpression of HPV DNA and ADF was observed in all HPV DNA-positive cases. Coexistence of HPV DNA and ADF immunopositivity in neoplastic squamous cells of the cervix suggests that ADF expression closely reflects the intracellular event on HPV DNA replication.

In an open study the concentrations of azithromycin in plasma, urine, peritoneal fluid and gynaecological tissue in 20 patients undergoing elective gynaecological surgery were compared. Patients were allocated to one of four groups and all patients received a single 500 mg oral dose of azithromycin prior to surgery. In Group I, the dose was administered 24 h before surgery. In Groups II, III and IV it was administered 48, 72 and 96 h, respectively, prior to surgery. A total of 19 patients completed the study; one patient had peri-operative complications and did not proceed to surgery. High concentrations of azithromycin were found in gynaecological tissue up to 96 h after administration. The mean maximum observed concentration 24 h after administration was 1.44 +/- 0.22 micrograms/g. Using all data, the depletion rate constant was 0.0104 h-1, equivalent to a half-life of approximately 67 h. The mean concentration of drug in peritoneal fluid was approximately 9% of the mean concentration in gynaecological tissue. Tissue and peritoneal fluid azithromycin concentrations were much higher than plasma levels at the time of surgery. Detectable plasma levels were only found in four patients from Groups I and II. Six percent of the total dose was excreted in the urine during the seven-day period after drug administration. The single dose of azithromycin was well tolerated by all the patients in this study and no treatment-related side effects or laboratory test abnormalities were seen. It is concluded that a single 500 mg oral dose of azithromycin produces high and sustained levels in gynaecological tissue up to 96 h after administration.

To investigate the relationship between the sex steroid receptor (estrogen receptor [ER] and progesterone receptor [PR]) status and the cell proliferation kinetics during the menstrual cycle in normal and neoplastic epithelium of the uterine cervix, immunohistochemical localization of ER, PR, and cell proliferation-associated antigen, Ki-67, was investigated in 35 normal cervical specimens, 3 condylomas, 26 cervical intraepithelial neoplasia (CIN) samples, and 22 invasive squamous carcinoma samples. The presence of human papillomavirus (HPV) DNA was also studied. In the normal cervix, basal cells were usually ER positive, PR negative, and Ki-67 negative throughout the menstrual cycle. Parabasal cells were ER positive and PR negative in the follicular phase, but ER negative and PR positive, and Ki-67 positive in the luteal phase, and Ki-67-positive cells increased in number in the luteal phase. In contrast, PR positivity was observed in the cells of condyloma (2 of 2 cases), CIN (19 of 26 cases), and invasive squamous carcinoma (13 of 22 cases) irrespective of the menstrual phase. Moreover, most neoplastic cells containing HPV DNA type 16/18 were ER negative, whereas several lesions containing HPV DNA type 31/33/35 were weakly ER positive. Many Ki-67-labeled cells were observed in the neoplastic lesions. These results suggest that reduced ER expression and increased PR expression are associated with the proliferation of normal cervical squamous epithelium, and this proliferation-related receptor status, which is probably induced by HPV infection, is usually expressed in neoplastic cervical squamous cells.

Background: Preterm delivery is the leading cause of neonatal mortality in the United States, but efforts to address the problem are hampered by the inability to predict accurately which pregnancies are at risk. We postulated that damage to the fetal membranes may release fetal fibronectin into the cervix and vagina, giving rise to a biochemical marker for preterm delivery., Methods: We measured fetal-fibronectin concentrations in cervical and vaginal secretions, amniotic fluid, and maternal plasma with a sensitive immunoassay using the monoclonal antibody FDC-6. Immunohistochemical studies were used to determine the distribution of fetal fibronectin in the placenta and amniochorionic membranes and to ascertain its cell of origin., Results: Women with uncomplicated pregnancies (n = 163) who delivered at term rarely had cervicovaginal fetal-fibronectin concentrations above 0.05 micrograms per milliliter between 21 and 37 weeks of gestation (11 of 267 cervical samples [4 percent] and 9 of 267 vaginal samples [3 percent]. High levels of fetal fibronectin were detected in amniotic fluid and in the cervical or vaginal secretions of 93.8 percent of the women with preterm rupture of membranes (n = 65). Cervical or vaginal fetal fibronectin was also present in 50.4 percent of the women with preterm uterine contractions and intact membranes (n = 117), and its presence identified the women who delivered before term (n = 60) with a sensitivity of 81.7 percent and a specificity of 82.5 percent. In the placenta and membranes, fetal fibronectin was found at points of contact with the uterine wall., Conclusions: The presence of cervicovaginal fetal fibronectin in the second and third trimesters of pregnancy identifies a subgroup of women who are at high risk for preterm delivery. This phenomenon may reflect the separation of the chorion from the decidual layer of the uterus, with the release of intact or degraded chorionic components of the extracellular matrix into the cervical and vaginal secretions.

Cervical scrapes from 43 patients (12 with chronic cervicitis, 13 with dysplasia, 11 with carcinoma in situ and 7 with invasive carcinoma) were examined by the polymerase chain reaction (PCR), to detect the amplified E7 gene of the HPV types 16 and 18 DNA sequence. HPV types 16 and/or 18 DNA were detected in 25 of 43 cases by the PCR (type 16 in 16, type 18 in 8, and both types 16 and 18 in 1). In comparison with the results for histological grades, the positive rate significantly increased as the grade of cervical dysplasia became higher. All cases were also examined with a HPV detection kit, the "Vira Pap" which contains 32P-labeled mixed RNA probes complimentary to HPV types 6, 11, 16, 18, 31, 33, 35 DNA. Six of 18 cases in which HPV DNA were not detected by the Vira Pap were positive for HPV types 16 and/or 18, so that the PCR was a highly sensitive method compared to the Vira Pap. The PCR has some advantages: 1. it requires only a small amount of specimens and 2. paraffin-embedded sections can also be used. We suggest that the PCR is a useful method for the screening and for retrospective investigation of HPV infection.

Paraffin-embedded tissues from a variety of sources and treated with different fixatives were tested for beta-globin and HPV amplification by polymerase chain reaction (PCR). In tests of tissues collected in the previous 2 to 3 yr, excellent rates (87% to 93%) for beta-globin amplification were obtained for specimens fixed in buffered formalin, Bouin's, and Hartmann's solutions. In contrast, the rate of beta-globin amplification was low for tissues fixed in Hollande's solution (7%) and in Hartmann's solution with eosin (33%). The results of beta-globin amplification from archival tissues stored for variable time periods showed no decrease in the amplification rate with longer periods of storage. Human papillomaviruses (HPVs) were identified in 17% of globin negative and in 43% of globin positive tissues. HPV-16 amplification was more efficient when the targeted DNA sequence was small. Variability in amplification depends not only on the type of fixative used, but also on other ill-defined factors. Therefore, conditions for optimal amplification should be determined before undertaking studies of archival material.

50 colposcopic biopsies of cervical epithelium were studied, using a silver colloid technique. These comprised 28 cases of human papillomavirus infection of the cervix, 8 cases of cervical intraepithelial neoplasia (CIN) I, 8 cases of CIN II, 6 cases of CIN III. The AgNOR mean number of the basal and parabasal cells of the cervical epithelium was significantly different in virus infected cells and in CIN. Different patterns of AgNOR distribution were observed: they were single and compact in virus infection without dysplasia whereas they appeared small and often loosely arranged in dysplastic lesions. Our data suggest that this simple technique is diagnostically useful in the evaluation of borderline lesions.

Using a modified Feulgen hydrolysis procedure and integrating microdensitometry, the acid-labile nuclear DNA in exfoliated cervical epithelial cells was quantified in a range of histologically confirmed cervical intraepithelial neoplasia (CIN), invasive cancer, and normal controls. The mean relative optical densities obtained for each sample group showed an increase from normal epithelium, through CIN grades, to invasive cancer. Although there was some overlap between groups, the difference in the overall mean values between the adjacent groups was statistically significant. The sensitivity of the test was 87.1% with a specificity of 99.2% and a predictive value of 99.5%, with no false negatives in the severe dysplasia and cancer groups. Quantitative data allows the threshold value to be altered to vary the sensitivity and specificity according to prevailing requirements. This suggests the possibility of using quantitative acid-labile DNA measurements to improve existing screening for cervical precancer.

We assessed the maximal muscular contractile ability, the passive biomechanical properties, and the hydroxyproline concentration in uterine cervical tissue samples from 28 nonpregnant women. Circular cervical tissue strips were mounted in organ baths and isometric tension was recorded. The mean (+/- SEM) maximal mechanical responses induced at the length of optimal mechanical performance by K+ (124 mmol/L) equaled 0.16 +/- 0.05 mN/mm2 in the distal cervix and 0.84 +/- 0.47 mN/mm2 in the proximal cervix, as compared with 4.85 +/- 1.0 mN/mm2 in tissues from the isthmus and 6.50 +/- 1.4 mN/mm2 in the fundus. The passive biomechanical properties were analyzed by a materials-testing machine. No significant differences were found between tissues from the distal and proximal cervix or between circular and longitudinal preparations. Tensile strength equaled 1.5-1.7 N/mm2, ie, 10(4)-fold the maximal muscular contractile ability. The extensibility was 0.63-0.76 and the stress-relaxation was 41-48%. The hydroxyproline concentration was 22.5 micrograms/mg wet weight in the distal cervix and 21.6 micrograms/mg in the proximal cervix, as compared with 16.6 micrograms/mg in the isthmus and 12.6 micrograms/mg in the fundus. A method for analysis of the biomechanical properties of the human cervix was designed, and from the measurements obtained it may be concluded that the passive biomechanical strength of the cervix markedly exceeds the active muscular contractile ability. This may be explained by a high collagen concentration and a low content of smooth muscle in the cervical tissue.

Human papillomaviruses (HPVs) are associated with benign and malignant neoplasms of the cervix. One of the criteria for their etiologic role requires an assessment of whether virtually all or only a small fraction of lesions contain viral genomes. DNA preparations from colposcopically directed punch biopsies of cervical lesions were analyzed by Southern blot hybridization and the polymerase chain reaction (PCR) for the presence of HPV DNA. The biopsy specimens represented different pathologic entities (koilocytosis, condyloma, cervical intraepithelial neoplasia, and invasive carcinoma). In Southern blot hybridization with radioactive probes for HPV 11, 16, 18, 31, and 33, HPV DNA was detected in 74% of the biopsy specimens (42 of 57 cases), with the predominant types being HPV 16 and HPV 18. In contrast, after PCR amplification with primers yielding fragments of characteristic size for HPV 11, 16, and 18, the analysis of the same 57 biopsy specimens revealed that all samples were positive for at least one HPV type. To exclude false-positive PCR results, controls without HPV DNA were interspersed at regular intervals, and results were evaluated only if these controls remained HPV negative. To exclude false-negative results due to failure of the reaction, a target sequence within the c-Ha-ras-1 gene was used as an internal control. All HPV typing results obtained by Southern blot hybridization were in agreement with HPV typing by PCR. The higher number of positive samples in the latter analysis stems from the increased sensitivity of PCR, which was which was effective in identifying as few as 10-100 HPV DNA molecules; in contrast, the sensitivity of Southern blot hybridization was 1 pg, or approximately 10(5) molecules of HPV DNA. The authors conclude that, with sufficiently sensitive diagnostic methods, HPV DNA can be detected in most, if not all, neoplastic cervical lesions.

CA 19-9, a side branch of the Lewis blood group system, is a sialylated Lewis A antigen that is highly expressed by many adenocarcinomas of the digestive tract. The müllerian duct-derived mucosa of the uterus and fallopian tubes also synthesizes Lewis blood group antigens. To test whether the expression of CA 19-9 is enhanced in carcinomas of müllerian duct origin, we performed immunohistochemical staining for CA 19-9 in normal tissues from 33 women and in adenocarcinomas from 88 patients. In the normal uterine cervix, CA 19-9 was expressed in the cytoplasm of scattered glandular cells in 26 of 29 specimens. It was observed in the apical regions of mucosal cells in six of 26 normal endometrial samples and two of 13 normal fallopian tube specimens. These results are consistent with the presence of antigen CA 19-9 on a secretory product of the nondiseased mucosa of the müllerian duct. In adenocarcinomas of the endocervix, endometrium, and fallopian tubes, CA 19-9 was found in seven of 11, 57 of 71, and five of six samples, respectively. Progressive loss of differentiation was accompanied by disruption of subcellular localization of CA 19-9 and its secretion toward the glandular lumina. In well-differentiated regions of tumors, the antigen was detected mainly at the luminal surface of cancerous glands, whereas the staining was mostly cytoplasmic or vacuolar in less differentiated areas. The degree of CA 19-9 expression was inversely related to tumor differentiation (P less than .001).

A chemiluminescent-labeled DNA probe (Gen-Probe PACE 2 System) for detection of Chlamydia trachomatis in endocervical specimens was evaluated. For each specimen, tissue cell culture, direct immunofluorescent staining (DFA), and DNA probe assay were performed. Thirty of 318 (9.4%) specimens were positive for C. trachomatis. The sensitivities, specificities, and positive and negative predictive values of the DNA probe compared with cell culture were 93%, 98%, 85%, and 99%, respectively, and for DFA these same values were 81%, 99%, 83%, and 99%, respectively. The Gen-Probe PACE 2 System is a reliable method for the rapid detection of endocervical chlamydial infection.

One hundred forty-five nonsmokers found to have normal cytologic diagnoses on routine Pap smears were interviewed regarding environmental exposure to tobacco smoke, and a 3 ml saline lavage of the cervix was collected for measurement of cervical nicotine levels by gas chromatography-mass spectroscopy. Nicotine levels tended to be highest among women exposed to tobacco smoke in the home, intermediate in women exposed only outside the home, and lowest in women recalling no exposure (p = 0.001).

The polymerase chain reaction (PCR) DNA amplification method is a powerful new tool for the retrospective analysis of paraffin-embedded tissue (PET). The technique has afforded the sensitive and specific detection of nucleic acid sequences associated with genetic and infectious diseases. However, PET processing conditions vary in their suitability for amplification. The authors have examined the effects of 11 fixatives at three fixation times. The effect of fixation was measured by the ability of the DNA in a treated tissue to serve as a template for the amplification of DNA fragments that ranged from 110 to 1,327 base pairs in length. Specimens fixed in acetone or 10% buffered neutral formalin were found to be best suited for subsequent analysis by PCR. A second group of fixatives, including Zamboni's, Clarke's, paraformaldehyde, formalin-alcohol-acetic acid, and methacarn, compromised amplification efficiency. Tissues treated with Carnoy's, Zenker's, or Bouin's, respectively, were even less desirable for amplification analysis.

The uterine cervical tissue was homogenized in STM buffer (0.25 M sucrose, 10 mM MgCl2, 10 mM Tris, pH 7.5). A soluble fraction with cytoplasmic proteins was obtained in the supernatant by centrifugation at 100,000 g for 40 minutes. According to the Davis method, there was almost no difference between the most abundant native cytoplasmic proteins (0.8/0.4/0.1/0.3) and the control serum proteins (0.8/0.1/0.4/0.3). Otherwise, according to the Laemmli method, the third and the fourth number of the "four-part numerical code" were strongly different for cytoplasmic proteins (67, 41, 186, 294) toward serum proteins (67, 41, 112, 235). Those data confirm Irwin's suggestion about the specificity of the "tissue codes".

Until now, efforts to automate cervical smear diagnosis have focused on analyzing features of individual cells. In a complex specimen such as that obtained from a cervical scrape, diagnostically significant cells may not be adequately represented or may elude detection by the automated technology. An approach is needed that extracts additional quantitative information from cervical smears beyond what the cell-by-cell approach can provide. A new methodology, contextual analysis, was developed to extract global quantitative information about cells, cell clusters, and background debris. This pilot study was designed to compare the efficacy of contextual analysis with high-resolution, single cell analysis and the analysis of intermediate cell markers. Thirty-four samples prepared as monolayers and stained with the Feulgen-Thionin/Congo Red stain were measured. Contextual analysis alone was able to classify 91% of the smears correctly; single cell analysis classified 94% of the cells correctly; and the intermediate cell analysis correctly identified the smear diagnosis for 84% of the cells. When all three analysis methods were combined into a simple smear level classifier, the overall smear classification accuracy was improved over those obtained using the three methodologies alone.

Cytomorphology, mucin stains, and immunocytochemistry were used to determine the characteristics of intracytoplasmic vacuoles in cervical smears and their relationship to chlamydial infections. Three study groups were examined: A) cervical smears containing intracytoplasmic vacuoles suggesting chlamydial infection: B) cervical smears from patients known to be either culture positive or culture negative for chlamydia; and C) control smears of chlamydia-infected McCoy cell cultures. Forty-nine smears containing vacuoles suspected of being chlamydial inclusions were studied. In 21 of 31 smears the vacuoles were shown by histochemistry to contain mucin. Immunoperoxidase staining performed on the remaining 18 smears revealed only two vacuoles to be chlamydial inclusions. Of 38 smears from chlamydia-culture-positive patients, only 4 showed chlamydial inclusions by immunoperoxidase methods. Of 30 smears from chlamydia-culture-negative patients, none were positive by immunoperoxidase staining. The "nebular body" of Shiina appears to be the most specific cytomorphologic evidence of chlamydial infection in cervical smears, but distinction from mucin vacuoles and other nonspecific vacuoles is difficult. The immunoperoxidase technique is unlikely to be useful in diagnosis in cervical smears unless samples are specifically obtained from the endocervical canal by endocervical brushing.

Epithelium transformation zone in the uterine cervix was studied by means of monoclonal antibodies (MAB) against cytokeratin (CK) peptides, epithelial membrane antigen (EMA) and skin granular layer proteins. It is shown that in the region of epithelial junction metaplastic epithelium is, as a rule, observed which possesses a unique spectrum of CK distinct from that in the ecto--and endocervix. The features of the expression of CK N8, N17, EMA in metaplasia are described. MAB EE 2I-06 against CK N1, 2, 9-11 and MAB G 36-19 are suggested as markers of the metaplastic epithelium maturity in the uterine cervix.

Progesterone receptor (PR) expression was studied in human cervix uteri from 21 cases. The study was performed on fresh frozen material with application of mouse anti-rabbit PR antibody and the immunoperoxidase method. Most cervical PR-s were present in the glandular epithelium and in smooth muscle cells. The results suggest that PR expression in the cervical glands is hormone-dependent; it was strongest in the presence of estrogen-stimulated endometrium, weaker with progesterone stimulation and absent in atrophy. The ectocervical epithelium contained relatively few progesterone receptors.

Relaxin was determined in healthy and pathological tissue samples from mammary gland, ovary, myometrium and cervix uteri of non-pregnant women. Relaxin could be detected in all groups of tissues examined. The highest values were found in healthy ovary, the lowest concentrations in the cervix, which is known as a target organ for relaxin activity during pregnancy. Mammary parenchyma showed the highest specific activity of all healthy tissues examined. The relaxin concentration in myoma tissue is significantly higher than in healthy myometrium.

To investigate the estrogen receptor (ER) status of cells during carcinogenesis of the uterine cervix, the immunohistochemical reactivity for a monoclonal anti-ER antibody (H 222) was studied in 26 normal cervical specimens, 21 cases of cervical intraepithelial neoplasia (CIN), and 21 cases of invasive cervical carcinoma. In addition, the presence of human papillomavirus (HPV) DNA (types 6/11, 16/18, or 31/33/35) was analyzed by in situ hybridization. In the normal cervix, basal cells of the squamous epithelium, metaplastic cells, and endocervical glandular cells were ER positive. In contrast, neoplastic cells of CIN (17 of 21 cases) and invasive carcinoma (19 of 21 cases) were ER negative. The remaining four cases of CIN and two cases of invasive carcinoma were focally ER positive. The HPV DNA analysis revealed that HPV DNA in ER-negative cases was either types 16/18 or undetectable, but all ER-positive neoplasms contained HPV DNA types 31/33/35. These results suggest that most neoplastic cells in CIN and invasive cervical carcinoma lose their ER expression and that this may be related to the HPV DNA types which they possess.

Although interferon-gamma has been associated with control of chlamydial infections in mice, no direct evidence links it to human chlamydial infections. Therefore, interferon-gamma was assayed by ELISA in endocervical secretions and plasma of women cultured for Chlamydia trachomatis. Women with positive endocervical chlamydial cultures had increased levels of interferon-gamma in endocervical secretions (6.7 +/- 2.8, mean +/- SE, n = 47) compared with uninfected women (1.4 +/- 0.4, n = 52) (P = .002). Interferon was also present in secretions of women with gonorrhea. Higher levels were seen in secretions from older women with positive chlamydial cultures. Interferon levels in secretions did not correlate with simultaneous plasma levels, the number of organisms recovered in tissue culture, or clinical correlates of inflammation. These data suggest that interferon-gamma is present at the site of chlamydial infection; however, further experiments are needed to determine whether interferon is specifically involved in protection or is a nonspecific indicator of inflammation.

The presence of chlamydial deoxyribonucleic acid (DNA) was evaluated by DNA hybridization in endocervical cells of infertile and normal fertile women. Chlamydial DNA was detected in 49 of 186 (26.3%) infertile patients, which is significantly more common than in fertile control individuals (12.5%, or 8 of 64 individuals). Among infertile patients, 49.3% (33 of 67) of those with tubal factors as cause of infertility and 13.4% (16 of 119) of those with nontubal factors were found to contain chlamydial DNA in their endocervical cells. The results show that chlamydial DNA could be found significantly more frequently in endocervical cells of infertile patients with tubal factor than those without tubal factors or in normal controls.

Quantitative micromethods have been used for measuring reactive protein thiols (PSHr), total reactive protein sulfur (TRPS), total protein thiols (PSHt), and protein disulfides (PDS) in fixed frozen sections of human uterine cervix. PSHr and TRPS were stained using 2,2'-dihydroxy-6,6'-dinaphthyl disulfide; PSHt and PDS were stained using mercurochrome methods. Microspectrophotometric measurements were made on the stained sections using a microdensitometer with associated data processing; the results obtained for areas of epithelium and stroma were converted to absorbance values per micron 2. Samples of uterine cervix that were diagnosed as containing cervical intraepithelial neoplasia (CIN) I-III or carcinoma were examined and compared with samples of normal uterine cervix. Measurements were made not only on identified lesions but also on apparently normal tissue obtained from the same cervix. Epithelial/stroma ratios (E/S) were calculated for PSHr, TRPS, PSHt and PSHt + PDS; in addition, the double ratios of PSHr/TRPS and PSHt/PSHt + PDS were also calculated for E/S. The mean E/S values for PSHr and PSHt were significantly different for all types of lesion compared with control samples. The E/S ratios for apparently normal tissue obtained from cervices with CIN or carcinoma were also significantly different compared with corresponding control values, indicating a field effect. There was a considerable degree of overlap between individual values in the control groups versus those obtained with each type of lesion. The corresponding mean E/S values for TRPS and for PSHt + PDS in the samples containing lesions were not significantly different from control means except for the group containing CaCx. However, the mean values for the double ratios (PSHr/TRPS and PSHt/PSHt + PDS) were significantly different in the groups containing lesions compared with the controls. Moreover, apparently normal tissue obtained from cervices containing CIN or carcinoma had different mean values compared with the controls, confirming the existence of a field effect. The degree of overlap of individual values in the lesion groups compared with the control values was much less with double ratio values than previously noted for single ratio values. In consequence, the double ratio measurements clearly discriminated CIN I + II and CIN-III from controls. Our data show that CIN is associated with marked changes in tissue protein thiols and disulfides and that these differences extend to neighboring apparently normal tissue indicative of a field effect.

In order to assess the burden of environmental pollutants in the genital tract, 12 different chlorinated hydrocarbons were determined in 152 samples of follicular, seminal and cervical fluids from sterility patients in the in vitro fertilization programme at the Centre for Obstetrics and Gynaecology of the University of Bonn. The lowest concentrations were found in the follicular fluids of patients who eventually became pregnant by in vitro fertilization. Higher concentrations were found in patients with sterility of unknown origin. Concentrations in the cervical secretion were up to twenty times higher than those in the seminal or follicular fluids. The results show that considerable concentrations of chlorinated hydrocarbons may be present in parts of the reproductive system, and that these compounds accumulate in the reception zone for spermatozoa. Certain pollutants therefore probably contribute to infertility.

Normal tongue and cervical mucosa, premalignant cervical and vulvar lesions, primaries and metastases of squamous cell carcinomas from the oral, laryngeal, cervical and vulvar mucosa were analyzed for c-erbB2 and c-myc transcription with northern-blots using 32P single-stranded RNA probes. Transcription of c-erbB2 and c-myc could be detected for almost all tissues including normal samples. A slightly enhanced transcription level was found in three cervical intraepithelial neoplasias of Grade III (CIN III) but in none of the malignant lesions. Increased transcription of c-myc was observed in premalignancies and malignancies. It was more frequent in oral and laryngeal squamous cell carcinomas (SCC) (8 of 9 cases) than in genital SCC (3 of 11 cases) or premalignancies (3 CIS of 14 CIN/VIN). No relationships of c-myc enhanced transcription level with tumor grading and staging were noticed. Thus, mere oncogene expression is a widespread phenomenon in tissues of different histogenesis and quantitative analysis is necessary prior to ascribe any diagnostic or prognostic relevance. Moreover, the frequency of tumors with enhanced transcription may vary for phenotypically closely related tumors of different organs.

This report emphasizes the ability to quantify ascorbic acid (AA) and reduced glutathione (GSH) levels in exfoliated cervicovaginal epithelial cells obtained by a lavage technique. Sixty-two women with abnormal Papanicolaou smears underwent colposcopic examinations. Colposcopic lesions were biopsied and histopathologically graded. Marked variations in the number of cells and in the levels of AA and GSH were observed. In cigarette smokers, the number of exfoliated cells retrieved was significantly higher (p less than 0.05, by Student's t test). The simultaneous investigation of biochemical and virologic parameters in exfoliated cervicovaginal epithelial cells, in conjunction with the known cytopathologic and epidemiologic risk variables, provides a novel approach to elucidate factor(s) that may inhibit or promote cervical carcinogenesis in designed prospective studies.

The cervical cytologic screening is responsible for the overall decline in the incidence of cervical cancer. Despite this apparent success, the cytologic assessment by light microscopy of cervical smear has recently been shown to have deficiencies particularly in cervical intraepithelial neoplasia (CIN). Now, this pathology of the uterine cervix is increasingly becoming recognized as a major health problem in young women because of its high frequency and its association with the development of cervical carcinoma. Flow cytometry (FCM) is widely used as a rapid method to analyse DNA content of a great number of cells on a cell by cell basis. The authors have applied FCM to CIN in order to reinforce the cytologic screening of cervical smears. Three abnormal DNA patterns have been observed: increased and heterogeneous DNA pattern; increased S and G2+M phases; abnormal DNA content. The statistical calculation has emphasized the significant relation linking these DNA patterns with the cytologic diagnosis and the severity of CIN. Therefore, FCM is a powerful complementary tool in cervical pathology.