PCR detection of human papillomavirus of the mucosa: comparison between MY09/11 and GP5+/6+ primer sets.

BACKGROUND AND OBJECTIVES: Human papillomaviruses (HPV) are the causal agent for the development of carcinomas in the cervix uteri and further pathological changes of the skin including mucosa, particularly warts, condylomas and dysplasias. Therefore, we investigated the efficacy of different consensus primers pairs for HPV detection by PCR using brushed samples from the oral cavity in comparison with samples from the cervix uteri. STUDY DESIGN: In the present study, we used two well-established sets of PCR primers in different combinations for the detection of HPV DNA in 106 non-invasive brush biopsy samples of the oral mucosa and 56 samples from the cervix uteri. Direct sequencing of PCR products in all cases determined HPV genotype and specificity. RESULTS: Overall, HPV was detected in 69 of 106 oral mucosa samples. HPV specific amplicons were obtained in 35.8% (N = 38) when using GP5+/6+ primers. The positivity rate was increased to 65.1% in a GP5+/6+ auto-nested PCR approach. In contrast, MY9/11 PCR and nested PCR with MY9/11 outer followed by GP5+/6+ inner primers yielded 2.2% and 16.1%, respectively. In gynaecological samples, PCR results were similar independent of the primer combination used. Thus, DNA quality and DNA content could be additional factors influencing the rate of positivity. CONCLUSION: For oral mucosa samples, auto-nested GP5+/6+ PCR is in our hands the most suitable approach for epidemiological studies because of its high sensitivity, high reliability and reproducibility as well as its relatively simple laboratory procedure.

[Diagnostic verification of Group III and III D cytological tests with special reference to follow-up]. / Die diagnostische Klärung von Zytotesten der Gruppen III und III D unter besonderer Berücksichtigung von Verlaufskontrollen.

The diagnostic verification of Pap III and Pap III D-smears from mass screening is described with special regard to the importance of colposcopical and cytological long-term controls. 233 (59.8%) out of 390 patients were verified within one year (1.1.-31.12. 1977). A high percentage (27.5%) of the first control smears was negative or the cytological picture had become normal (19.5%) after vaginal treatment by antibiotics and application of estrogens. 50 women were cured by conization or primary hysterectomy as the cytological diagnosis yielded suspicion of severe dysplasia or carcinoma in situ. Depending on cytological differential diagnosis, individual situations and the developmental tendency of the atypical epithelium 157 patients (40.2%) were observed for several years. The great diagnostical-therapeutical importance of colposcopy for the early recognition of cervical intraepithelial neoplasia (CIN) is demonstrated. A direct microbiopsy controlled by colposcopy is a valuable completing method if there are differences between colposcopical and cytological findings during a long-term control of the cases. A slight or moderate dysplasia can be observed for about 2 or 3 years, as most regressions are seen in this period. In problematic cases (young nullipare, patients treated because of infertility or sterility, pregnant women, old women) with severe dysplasia/carcinoma in situ it is possible and justifiable to control the patient temporarily in short distances. All cases which are suspicious of an early stromal invasion call for a definite diagnosis and therapy without any delay. Primary hysterectomy is preferred in patients aged 35 years or more.

[Low temperature biopsy of biological tissues (author's transl)]. / Tieftemperaturbiopsie biologischer Gewebe.

A method and an equipment are described to perform a low temperature biopsy. The advantages of this method are the immediate conservation and the excellent topical conjugation of tissue. Using the effect of adherence it is also possible to get a biopsy from tissues of a high rupture strength. We used this cryobiopsy method for histological diagnosis of colposcopically suspicious areas of the ectocervix uteri, where the representation of relations between epithelium and stroma is of practical importance.

[Conisation and pregnancy (author's transl)]. / Konisation und Schwangerschaft.

Reference is made to experience obtained from specialised outpatient practice in an account of diagnostic and therapeutic approach to pregnant women with pathological cervix smear. Six out of 93 pregnant women were conisised during pregnancy and 41 after delivery. Postpartum primary vaginal hysterectomy was performed on three women with additional gynaecological indication. Strict follow-up checks were made on 43 patients, among them three with residual dysplasia who had undergone conisation prior to conception. Differentiated approach, which is of great practical importance, in particular to high-risk pregnancy, has proved to be possible by specialised diagnosis, with colposcopy, differential cytological testing, and microbiopsy being included. Postponement of conisation to a date after delivery or after achievement of the desired family size can be justified under the condition of regular follow-up checks for women with intra-epithelial cervical neoplasia, on account of the latter's long latency. However, immediate therapeutic action, even within pregnancy, must be the response to any indication to invasive growth.

[Central neurotoxic side effects following cisplatin therapy]. / Zentrale neurotoxische Nebenwirkungen nach Cis-Platin-Therapie.

A 48 years old patient underwent a surgical treatment because of ovarian carcinoma developed different central neurotoxic events after chemotherapy with longer time known drugs as well as after Cis-platin-application.

[Passive electrical tissue parameters of normal and pathologically altered cervical epithelium]. / Passiv elektrische Gewebeparameter des normalen und pathologisch veränderten Portioepithels.

Normal and pathologically changed tissues can be distinguished on the basis of passive electric measurements, if the concentrations of the membranes, the resistance of the membranes, the membrane capacity, or the conductivity in and outside of the cells are different for both tissues. The electrical parameters are immediately measurable without anesthesia under in vivo conditions by change of electrical pulses. 2043 measurements on 300 patients were carried out. A suitable combination of electrical parameters of the tissues revealed that there is a difference of 64.5 +/- 0.5% between normal and pathologically changed epithelium (cancer).

[Pulse cytophotometric and cytologic repeated examinations of cervical secretions as compared with histologic findings (author's transl)]. / Impulszytophotometrische und zytologische Mehrfachuntersuchungen am Zervixsekret mit histologischen Vergleichen

1625 cervical smears from 397 women were investigated in special cytological consultations. On the same material 693 impulse cytophotometric (ICP) measurements were made, stained with ethidium bromide after pepsination. In 207 patients the diagnosis was certain by histology. The mean height of the 4c-peak (in 0/00 of the 2c-peak) raises with the Papanicolaou grading from 38,2 in Pap. I up to 128,8 in Pap. V. In histological confirmed cases in dysplasia the mean 4c-height was 79,5 and in carcinoma 102.2. If only one ICP investigation was turned to account there are some disagreements between the ICP value and the diagnosis finally resulting from the clinical and cytological course. The differences are considerably diminished by repeated ICP investigations. Therefore ICP measurements are a valuable aid in interpreting cytodiagnostic problems. They often facilitate the choice of further diagnostic and therapeutic procedures.

[Colposcopy and early changes of cervical carcinoma (author's transl)]. / Kolposkopie und Frühveränderungen des Zervixkarzinoms.

Cytology and colposcopy are well recognized methods in the early diagnosis of pre-malignant and early invasive changes of the uterine cervix. Since 1971 the University Department for Women in Leipzig has a cytology clinic. The results of repeat smears before biopsies and the follow-up examinations on 202 cone biopsies are reviewed. The incidence of negative findings decreases with increasing atypia of the squamous epithelium. The highest incidence of false negative smears was 73.3% in mild and moderate dysplasias. The more pronounced the atypia the smaller is the number of inadequate pap smears. In cases with negative cytology and colposcopic atypia the colposcopic findings must be clarified. Only the combination of colposcopy and cytology leads to optimal early diagnosis of cervical intra-epitheal neoplasia and invasive carcinoma of the cervix. The false positive cytosmears in the screening are discussed and a treatment with estrogens prior to repeat examination is recommended. Diagnostic problems arise from a discrepancy in the colposcopic and cytologic findings. Discrepancies depend on the colposcopic experience of the examiner. Improvement of the colposcopic diagnosis is obtained by directed colposcopic biopsy.

[Therapy outcome in stage III (FIGO) ovarian carcinomas at the Leipzig University Gynecologic Clinic 1976-1982 and 1985-1991]. / Therapieergebnisse bei Ovarialkarzinomen im Stadium III (FIGO) an der Universitäts-Frauenklinik Leipzig von 1976-1982 und 1985-1991.

This retrospective study compares treatment results (median survival time, 5-year survival rate) of 153 patients diagnosed with FIGO stage III ovarian cancer treated between 1976 to 1982 (n = 73) and 1985 to 1991 (n = 80) respectively. Between 1976 to 1982, the median survival time was 12.3 months, while it increased up to 17.0 months during the period of 1985-1991. The 5-year survival rate increased from 14.0% to 28.0%. This improvement may be due to a more radical surgical approach and the use of platinum-based first-line chemotherapy. However, the difference between the two groups was not statistically significant.

[Results of a psycho-oncological project at the University Women's Hospital Leipzig]. / Ergebnisse eines psychoonkologischen Projektes an der Universitäts-Frauenklinik Leipzig.

This paper deals with a synoptic description of experiences of an inter-disciplinary oncologic project by means of its three main components--theoretical research, further education and qualification as well as psychological intervention program. It was shown that team work between physicians, nurses and psychologists led to an new insight and higher working effectivity with minimal subjective stress. This team work led also to the creation of favourable conditions for an appropriate management of diseases through an intensive and integrated patient care. The application of this model project in the routine oncological follow-up would be recommended.

[Impulse cytophotometric studies in cytologically suspicious cervical smears]. / Impulszytophotometrische Untersuchungen bei zytologisch suspekten Zervixsekreten

187 women with cytologically suspicious cervical smears (Papanicolaou III to V) were investigated by 387 pulse cytophotometric (ICP) measurements. Within the Papanicolaou III group a significantly higher 4c-peak was seen in cases with only one ICP-investigation if we had histologically pathological findings (dysplasia, carcinoma) as contrastes with clinically and cytologically regressive cases. In repeated ICP-investigations we found in group Papanicolaou III (histol.: dysplasia, carcinoma) as well as in group Papanicolaou IV and V a significantly higher mean 4c-height than in only once investigated cases. Within a ICP-series the maximal 4c-height is most representative. The security of diagnosis is raising with repeating ICP-investigations. Here it is a valuable diagnostic aid especially in suspicious cases. All abnormal ICP-findings must be controlled for a long time, even with normal cytological smears. At last, the circumstances causing false negative ICP-results are discussed.

[Biological validity, cytodiagnosis, and therapy of cervix dysplasia]. / Zur biologischen Wertigkeit, Zytodiagnostik und Therapie der Zervixdysplasien

Certain forms of dysplasia are biologically equal to carcinoma in situ and should be treated alike. The persistance of the state during the permanent cytologic control is to be regarded as criterion for the treatment of middle and even the slight dysplasia. Slides of cells indicating a dysplasia are therefore placed in a cytologic risk-group III D. If the restoration of the cytologic state cannot be observed within 12 months, the conisation as carcinoma prophylaxis of slight and middle forms of dysplasia should be performed.

Predictive value of DNA cytometry in CIN 1 and 2. Image analysis of 193 cases.

OBJECTIVE: To investigate DNA image cytometry for predicting the prognosis of cervical intraepithelial neoplasia (CIN). STUDY DESIGN: Smears from 151 women affected by CIN 1 or 2 on cytology with minimal follow-up of three years were included. Sixty-seven showed progression, with histologically confirmed carcinoma in situ or invasive cancer. Eighty-four cases showed regression of the disease, which was cytologically, histologically and colposcopically confirmed. Papanicolaou-stained smears were destained, and the Feulgen reaction was performed with consecutive image DNA cytometry of suspicious cells using an image analysis system (Cires, Zeiss, Germany). The DNA index of the greatest stemline and the number of single aneuploid cells, using 9c exceeding events, were computed. RESULTS: In the group with progression, an aneuploid DNA stemline was found in 25 smears (26.9%). In 64 cases (66.7%) more than one aneuploid event was detected. The total number of aneuploid cases in this group was 76 (81%). In the group without progression, the number of aneuploid stemlines was 2 (2%). Single aneuploid cells could be found in five cases (5%). The overall number of aneuploid cases in that group was five. The sensitivity was 74.3%, positive predictive value 85.2% and negative predictive value 77%. CONCLUSION: Aneuploidy is a marker for prospective malignancy in cervical Papanicolaou smears. DNA image cytometry, as an additional method, can be used to predict outcome in patients with CIN 1 and 2 of the cervix. DNA cytometry is not a screening method but can add further information for a treatment decision in doubtful cases.

[Histopathologic correlates of false-positive cytologic findings in the uterine cervix]. / Histopathologisches Korrelat falsch-positiver Zytologiebefunde der Cervix uteri.

Between 1984 and 1992, we reinvestigated 170 cases of cervical smears obtained in conization or hysterectomy which had produced false positive cytologic findings. We looked for morphological changes that could explain the cytologic findings. Such changes were found in half of the cases. These cases included 54 (63.6%) specimens with marked cellular polymorphism as a result of inflammation, tissue regeneration, or atrophy. In 51.9% of the cases showed histologic signs of possible HPV infection. The initial findings required reclassification in 12.9% of the cases, and in fifteen cases (17.6%) the biopsy matched the cytologic findings, without being validated by the final findings. We observed histologic phenomena corresponding to the cytologic findings in 30.8% of the cases in group IIID (n = 26) using Papanicolaou's classification system. In groups IV a (n = 72), IV b (n = 44), and V (n = 28), we found a histologic explanation in 54.2%, 47.7%, and 60.7% of the cases, respectively. In most cases we observed polymorphic cellular changes primarily associated with HPV.

False positive cervical smears: a cytometric and histological study.

The evidence of exfoliative cytology of the cervix uteri depends on various factors. The rate of false positive cervical smears is on average 8.8%. We have reviewed 170 false positive routinely Papanicolaou-stained cervical smears and the corresponding histological specimens. The aim was to look for non-neoplasic changes explaining the cytological over-interpretation. In addition, nuclear Feulgen-DNA measurements of the smears were made to look for changes in DNA pattern. In 50% of all cases, a cause was found for the false positive cytological diagnosis by means of histological reexamination: non-neoplastic nuclear and cellular polymorphia, severe purulent-erosive inflammation, a higher reclassification or a positive presurgical biopsy. In 51.9% of these cases, there were signs of an infection with human papilloma virus (HPV). In cases without histological explanation of the cytological findings, DNA measurements were made on the suspicious cells. By means of single cell algorithm (5c/9c exceeding events), a general rate of 62.7% of aneuploid specimens was detected. The number of aneuploid single cells ranged between 5 and 63 per slide. Of these cases, 23.1% showed signs of HPV infection. As a result of the study, the number of real 'false positive' cytological diagnoses decreased from 170 to 97 as a result of histological reexamination and DNA measurement. In doubtful cases, DNA measurement of suspicious smears should be made.

[Cytodiagnostic studies of the laryngeal mucosa in chronic laryngitis]. / Zytodiagnostische Untersuchungen an der Kehlkopfschleimhaut bei chronischer Laryngitis.

Cytological diagnostic aids in chronic inflammation of the larynx have been rarely employed so far. A study was conducted to prove whether there is any possibility of larnygeal cytological diagnosis and whether there are any premalignant changes. In view of the results obtained by gynaecological exfoliative cytology, attention was focussed on the metaplastic cells. The frequent incidence of metaplasias was highly significant, whereas dysplasias occurred only in chronic laryngitis besides in tumours. One trend occupying higher cytological diagnostic groups in the Papanicolaou pattern was seen in chronic laryngitis. In our opinion, cytological diagnosis in laryngeal investigation is an aid in the followup control in chronic laryngitis, which is a repeatable procedure without unduly stressing the tissue. Recording of cellular changes in chronic laryngitis can enable a prospective appreciation of morphological changes, which can be useful as diagnostic tool for examining suspicious findings.

[Early diagnosis of relapses of cervical and endometrial carcinomas (author's transl)]. / Frühdiagnostik von Rezidiven des Zervix- und Endometriumkarzinoms.

A systematic screening-programme for tumor recurrence diagnosis was carried out in 773 patients with cervical or endometrial carcinomas. In comparison with two other patient groups in this patients an earlier diagnosis of tumor recurrences was possible and more relapses were detected in the subclinically symptomless phase. Considering the primary therapy a practicable scheme for diagnosis is proposed.

Transplantation of stem cells of embryonic liver in a patient with severe combined immunodeficiency.

A patient had severe combined immunodeficiency syndrome and X-chromosomal recessive heredity. Since the parents and siblings were not suitable as HLA-compatible bone marrow donors, stem cells from embryonic liver were transplanted intravenously in 3 stages (6 X 10(6); 3.5 X 10(6), and 9 X 10(7]. Transplantation was tolerated well; there were no signs of a graft-versus-host reaction. Examination of the immunological condition after transplantation showed evidence of T-cell reconstitution, immunohistochemistry revealed beginning immune globulin production. The child died at the age of 5 months due to respiratory failure.

[Results of cytological and histological studies on the uterine cervix after long-term intake of hormonal contraceptive agents]. / Ergebnisse zytologischer und histologischer Untersuchungen der Cervix uteri nach längerer Einnahme hormonaler Kontrazeptiva

Cytologic examinations with 880 women having constantly used oral contraceptives from one to eight years show no agglomeration of epithelial atypias if the primary status was clearly in the negative. Among 669 histologically sure atypias of cervix uteri the part of women using oral contraceptives for at least one year is significantly lower in carcinoma in situ as well as in early invasive carcinoma. The differences in dysplasia are not significant. The results are also discussed with regard to the efficiency of permanent control examinations before and during the use of oral contraceptions. The demand of a further intensification of cytodiagnostic is derived from these examinations.

PIP: Studies of the cervical cytological effects of oral contraceptives are reported. Precontraception cytology (all clearly Group 1 or 2) in 880 women was compared with cytological findings after 1-8 year (average 2.4 years) of continuous use of Ovosiston or Non-Ovlon, and with those of a control group of 479 women of similar age. In addition, 669 women with cervical pathology (dysplasia, preinvastive and invasive carcinoma) were questioned on previous use of hormonal contraception for at least 1 year. No evidence could be found that hormonal contraception fostered cytological atypias if initial findings were negative. The proportion of women with cervical pathology who had taken hormonal contraceptives was significantly smaller than their proportion in the ambulatory patient population. These results are discussed in relation to the need for prospective studies of the effects of oral contraceptives.