H. Schell

Papillomavirus DNA in warts of immunosuppressed renal allograft recipients.

SummaryRenal allograft recipients were shown to have an increased incidence of warts and skin cancers. We examined 148 patients for evidence of wart virus infections and tested for papillomavirus types, which are known to be associated with human malignancies. Of the 148, 36 (24.3%) patients were afflicted with warts at the end of our study period, in contrast to 5 of 148 (3.3%) before transplantation. DNA from 16 different biopsies was extracted by phenol treatment for further virological studies. DNA of human papillomavirus (HPV) 2 was detected three times, DNA of HPV 4 and 10 twice, and DNA of HPV 3 and 16 once each by blot hybridization. One probe led to strong signals with HPV types previously found only in epidermodysplasia verruciformis patients. A correlation between histology and virus type exsisted in cases of HPV 2, 3, 4, and 10 infections.

Effects of epidermal growth factor, fibroblast growth factor, minoxidil and hydrocortisone on growth kinetics in human hair bulb papilla cells and root sheath fibroblasts cultured in vitro

SummaryComparative studies on growth kinetics of cultivated human hair bulb papilla cells (PCs) and hair root sheath fibroblasts (RSFs) yielded evidence of some pecularities of PCs in both proliferative behavior and morphological growth pattern. As the dermal papilla, essentially supporting the nutrition of matrix epithelium, can be considered a target tissue for agents influencing maintenance of hair growth, we studied the effects of epidermal growth factor (EGF), fibroblasts growth factor (FGF), minoxidil (Mino), and hydrocortisone (HC) on the proliferation of PCs and RSFs, both gained from dissected hair follicles of scalp biopsy specimens of two male adults and separately cultured in vitro. EGF and FGF proved to increase proliferation of both PCs and RSFs most, yet at a different intensity for each cell group. HC slowed proliferation, and Mino failed to influence growth of PCs and RSFs.

Clinical relevance of sonometry-derived tumour thickness in malignant melanoma : a statistical analysis

High‐frequency sonography has been shown to be a useful tool in planning operative strategy in the surgery of malignant melanoma (MM). The purpose of the present study was to compare sonometric and histometric data of tumour thickness in primary cutaneous MM, applying statistical methods in order to evaluate the pre‐operative relevance of sonometry. The thickness of 259 melanomas was measured preoperatively by a 20‐MHz B scan, and postoperatively by histometry. Statistical analysis was performed using Pearsons correlation coefficient and absolute and relative differences. Although the correlation between sonometry and histometry was good (r=0·88), there was a mean difference of 0·39 mm (relative difference 28%). Overall, sonometry was in agreement with the corresponding histological classes in 75% of cases. However, tumours assessed by ultrasound as between 0·55 and 0·95 mm thick were incorrectly classified according to histology in 34%, and those between 1·30 and 1·70 mm were incorrectly classified in 50% of cases.

Comparative morphological and growth kinetics studies of human hair bulb papilla cells and root sheath fibroblasts in vitro.

SummaryDermal papillae isolated from anagen hair bulbs obtained from biopsy specimens from five subjects with normal hair pattern, and fibroblasts derived from the mesenchymal root sheaths (RSF) of the same hair follicles were separately grown in culture and the cellcycle distribution pattern on different days was analysed by applying DNA flow cytometry (FCM). Papilla cells (PC) exhibited distinctive morphological features by forming cell aggregates differing from RSF with respect to cell shape and growth pattern. They also proliferated remarkably more slowly than RSF. DNA-FCM analysis showed that both PC and RSF demonstrated synchronous fluctuations in the percentage of cells in G1/0, S and G2+M phases during the period of subculture.

Collagen-type synthesis in human-hair papilla cells in culture

SummaryHair-papilla derived cells were grown in monolayer culture and revealed the typical morphology and growth pattern which was similar but not identical to control fibroblasts. Hair-papilla cells were found to produce considerable amounts of collagen type I and type III and fibronectin. Type IV collagen production could not be detected. The ratio of collagen type III and type I clearly differed from the pattern observed in normal fibroblasts, being much higher in hair-papilla cells, where type III accounted for more than 20% of total collagen synthesis. These data show that hair-papilla derived cells have biosynthetic capacities similar to those of human skin fibroblasts as well as characteristic differences, indicating that they represent a specialized fibroblast subpopulation.

13-cis-retinoic acid in conglobate Acne. A follow-up study of 14 trial centers

SummaryA follow-up was done on the patients of the German multicenter study with severe conglobate acne who had been treated with different dosages of 13-cis-retinoic acid. Eighty-seven patients were monitored from 12 to 21 months. Optimal long-term therapeutic effects were obtained with an initial dose of 1.0 mg/kg body weight, for 3 months, followed by another 3-month-treatment period with 0.2 mg/kg body weight. Six months after the termination of therapy 96% of the patients were still in remission and 81% after 12 months. Comparative figures for the administration of doses were 84% as opposed to 47% (0.5 → 0.2 mg/kg body wt.) and 74% as opposed to 37% (0.2 → 0.2 mg/kg body wt.), respectively. It is suggested from the present data that a high initial dosage of 13-cis-retinoic acid be chosen in order to obtain optimal long-term therapeutic effects. Transiently elevated lipid levels as well as other tolerable side effects return to normal within 3 months at the latest after discontinuation of treatment.

Sex hormones and antiandrogens influence in vitro growth of dermal papilla cells and outer root sheath keratinocytes of human hair follicles

Anagen hair bulb papillae, interfollicular dermal fibroblasts, and interfollicular keratinocytes isolated from fronto-parietal scalp biopsies as well as outer root sheath keratinocytes from plucked anagen hairs were separately grown in subculture for 14 d. The effect of different concentrations (2.4 nM-17.3 microM) of testosterone, dihydrotestosterone, and the antiandrogens cyproterone acetate or 17 alpha-propylmesterolone on growth behavior of the mesenchymal and epithelial cell types of the hair follicle were comparatively studied by means of growth curves, cell doubling times, and 3H-thymidine incorporation. For control, all cell lines were subcultured in hormone-free medium. Testosterone and dihydrotestosterone (345 nM) significantly reduced proliferation of papilla cells compared with dermal fibroblasts (p < 0.01) and outer root sheath keratinocytes compared with interfollicular keratinocytes (p < 0.01), as well as compared with cells cultured in control medium. Low concentrations of 17 beta-estradiol were ineffective, whereas doses of 180 nM 17 beta-estradiol increased the growth velocities of all cell types, especially of papilla cells, compared with dermal fibroblasts. Low doses of either cyproterone acetate (24 nM) or 17 alpha-propylmesterolone (29 nM) induced a growth enhancement, especially of papilla cells and outer root sheath keratinocytes, whereas high doses of cyproterone (1.20 microM) and 17 alpha-propylmesterolone (1.45 microM) had opposite effects. These changes were significant between papilla cells and dermal fibroblasts as well as between outer root sheath keratinocytes and interfollicular keratinocytes. Applying increasing doses of androgens to cyproterone acetate (24 nM)- or 17 alpha-propylmesterolone (29 nM)-containing media neutralized the growth-stimulating effect of antiandrogens, particularly in papilla cells and outer root sheath keratinocytes. However, minor differences between testosterone and dihydrotestosterone effects on cell growth were found. The data clearly demonstrate that the changes of in vitro growth of hair follicle cells depend on the concentrations of androgens and antiandrogens, as higher doses of both antiandrogens tested retarded the cell proliferation similar to testosterone or dihydrotestosterone. The papilla cells and outer root sheath keratinocytes reacted more sensitively to the hormones tested, thereby confirming the concept of a distinct androgen sensitivity of these specialized hair follicle cells.

Cell Cycle Kinetics of Human Anagen Scalp Hair Bulbs in Thyroid Disorders Determined by DNA Flow Cytometry

In both, 6 hyperthyroid and 6 hypothyroid patients as well as 10 healthy volunteers, cell cycle kinetics of dissected anagen scalp hair bulbs were determined by means of DNA flow cytometry (DNA-FCM). Compared with the healthy control group in patients with thyroid disorders striking differences of cell kinetic data were evaluated. In hyperthyroidism a significant increase (30%) and in hypothyroidism a significant decrease (15%) of S and G2+M phase cell percentages was found. The proliferation index (S+G2+M %) calculated revealed similar results. A correlation between the height of S phase percentages and plasma T3 levels was recognizable but could not be proven statistically. By means of DNA-FCM the study demonstrates for the first time the influence of thyroid hormones on in vivo cell cycle kinetics of human scalp hair bulbs.

Lokal rezidiviertes und metastasiertes malignes Melanom Langzeitergebnisse und Prognosefaktoren nach perkutaner Radiotherapie

Hintergrund: Die perkutane Radiotherapie wird oft nur als “Ultima ratio” bei Patienten mit fortgeschrittenem malignen Melanom (MM) eingesetzt. Unsere Studie wertet 20 Jahre klinische Erfahrungen bei fortgeschrittenem, rezidivierten und metastasierten malignen Melanom sowie Endpunkte und Prognosefaktoren aus. Patienten und Methodik: Von 1977 bis 1995 wurde bei 121 Patienten (56 Frauen, 65 Männern) von 2 917 Patienten des örtlichen Melanomregisters die Indikation für eine palliative Radiotherapie bei fortgeschrittenem malignen Melanom gestellt. Zu Beginn der Radiotherapie hatten elf Patienten inoperable Primär- oder Rezidivtumoren oder wiesen postoperativ Tumorreste (R2) auf (UICC IIB); 57 Patienten hatten Lymphknoten- (n = 33) oder In-transit-Metastasen (n = 24) (UICC III); 53 hatten Fernmetastasen (sieben M1a; 45 M1b) (UICC IV). Von der Erstdiagnose bis zum Beginn der Strahlentherapie vergingen im Mittel 19 (median: 18; Spanne: drei bis 186) Monate. Bei 77 Patienten wurde eine konventionell fraktionierte Radiotherapie und bei 44 Patienten eine hypofraktionierte Radiotherapie mit Einzeldosen von 2 bis 6 Gy und einer mittleren Gesamtdosis von 45 (median: 48; Spanne: 20 bis 66) Gy appliziert. Ergebnisse: Eine komplette (CR) und/oder partielle Remission (CR + PR) nach drei Monaten erzielten sieben (64%) bzw. alle Patienten im Stadium UICC IIB, 25 (44%) bzw. 44 (77%) im Stadium UICC III und neun (17%) bzw. 26 (49%) im Stadium UICC IV. Bei 25 (21%) Patienten war der Tumor progredient. Patienten, die eine CR erreichten, lebten länger (median: 40 Monate) als solche ohne CR (zehn Monate) (p < 0,01). Am Stichtag der Auswertung (31.12.1996) lebten noch 26 Patienten; davon waren bei Bestrahlungsbeginn sechs (55%) im Stadium UICC IIB, 17 (30%) im Stadium UICC III und drei (6%) im Stadium UICC IV (p < 0,01). Univariate Prognosefaktoren für CR und Überleben waren das UICC-Stadium bei Radiotherapiebeginn (p < 0,001), primäre Lokalisation im Kopf-Hals-Bereich und Gesamtdosis > 40 Gy (p < 0,05). Alter, Geschlecht und Histologie waren ohne Einfluß. In multivariater Analyse war das UICC-Stadium der einzige unabhängige günstige Prognosefaktor für CR und langfristiges Überleben (p < 0,001). Schlußfolgerung: Die perkutane Radiotherapie ist eine effektive Behandlung des malignen Melanoms selbst in fortgeschrittenen UICC-Stadien. Das UICC-Staging ist nicht nur initial, sondern auch bei Metastasen prädiktiv für das Erreichen einer initialen und langfristigen Tumorkontrolle. Prospektiv randomisierte Studien sollten den Stellenwert der adjuvanten Radiotherapie bei fortgeschrittenem und metastasierendem malignen Melanom überprüfen.Purpose: Radiotherapy (RT) is used as last resort for patients with advanced cutaneous malignant melanoma (MM). Herein our 20-year clinical experience is presented analyzing different endpoints and prognostic factors in patients with locally advanced, recurrent or metastatic MM. Patients and Methods: From 1977 to 1995, 2,917 consecutive patients were entered in the MM registry of our university hospital. RT was indicated in 121 patients (56 females, 65 males) for palliation in locally advanced recurrent and metastatic MM stages UICC IIB to IV. At the time of RT initiation, 11 patients had primary or recurrent lesions which were either not eligible for surgery or had residual disease (R2) after resection of a primary or recurrent MM lesion (UICC IIB); 57 patients had lymph node (n = 33) or in-transit metastases (n = 24) (UICC III), and 53 had distant organ metastases (7 M1a, 46 M1b) (UICC IV). The time from first diagnosis to on-study RT averaged overall 19 months (median: 18; range: 3 to 186 months). In 77 patients conventional RT and in 44 patients hypofractionted RT was applied with 2 to 6 Gy fractions up to a mean total RT dose of 45 (median: 48; range: 20 60 66) Gy. Results: At 3 months follow-up, complete response (CR) was achieved in 7 (64%), overall response (CR + PR) in all (100%) UICC IIB patients, in 25 (44%) and 44 (77%) of 57 UICC III patients, and in 9 (17%) and 26 (49%) of 53 UICC IV patients. Tumor progression during RT occurred in 25 (21%) patients. Patients with CR survived longer (median: 40 months) than those without CR (median 10 months) (p < 0.01). At the time of evaluation and last FU (December 31, 1996), 26 patients were still alive: 6 (55%) stage UICC IIB, 17 (30%) stage UICC III, and 3 (6%) stage UICC IV patients (p < 0.01). Univariate analysis revealed following prognostic factors for CR and long-term survival: UICC stage (p < 0.001), primary location in the head and neck, total RT dose > 40 Gy (all p < 0.05), while age, gender and primary histological subtype had no impact. In multivariate analysis, UICC stage was the only independent favorable prognostic factor for achievement of CR and long-term survival (p < 0.001). Conclusions: External RT provides effective palliation in advanced UICC stages. The UICC staging system is a good predictor of initial and long-term tumor response in metastatic MM. Prospective randomized trials using RT with or without adjuvant therapy for advanced MM are justified.

Autoradiographic in vitro studies on diurnal variation in human epidermal cell proliferation.

SummaryFive men with healthy skin ranging in age from 24 to 37 years were investigated for the presence of diurnal variation in epidermal cell proliferation by the in vitro3H-thymidine labeling technique. The following parameters were studied: The basal cell LI; the numbers of labeled basal and supra-basal cell nuclei as well as the total number of labeled nuclei situated above a basal membrane length of 100 µm.All autoradiographic parameters showed marked diurnal variations, the amplitudes of which differed individually. The basal3H labeling index always showed the highest diurnal variation. In all patients diurnal variations of the basal and suprabasal cell proliferation occurred simultaneously and in equal direction. However, each test person showed maxima and minima of epidermal DNA synthesis at different times. No synchronism could be found when comparing the individual variations of cell proliferation. Several possible reasons for the asynchronous in vitro behaviour of epidermal cells will be discussed.ZusammenfassungAn 5 hautgesunden Männern im Alter von 24-37 Jahren wurde durch in vitro-3H-Thymidinmarkierung geprüft, ob tageszeitliche Schwankungen der epidermalen Proliferation bestehen. Untersucht wurden folgende Parameter: der3H-Index der Basalzellkerne, die Anzahl markierter Basalzellkerne/100 µ Basalmembranlänge, die Anzahl markierter Suprabasalzellkerne/100 µ Basalmembranlänge und die Gesamtanzahl markierter Zellkerne/100 µ Basalmembranlänge.Sämtliche autoradiographische Parameter zeigten deutliche diurnale Schwankungen, deren Amplitude individuell variierte. Die ausgeprägteste tageszeitliche Variabilität wies in allen Fällen der basale3H-Index auf. Die im einzelnen beobachteten diurnalen Schwankungen der basalen und suprabasalen Proliferation traten gleichzeitig auf und verliefen gleichsinnig. Jedoch zeigte jede Versuchsperson zeitlich unterschiedliche Maxima und Minima der epidermalen DNS-Synthese, so daß keine diurnale Synchronizität der individuellen Proliferationsschwankungen nachgewiesen werden konnte. Einige mögliche Gründe für das desynchrone Verhalten der Epidermiszellen in vitro werden diskutiert.