[Immunomodulation by cryosurgery in malignant melanoma]. / Immunmodulation durch Kryochirurgie beim malignen Melanom.

Cryosurgery is a well-known, established method for the local destruction of tumor tissue by freezing. The assumption that, in addition to a physical and blood vascular phase, an immunological phase exists, has been discussed by many authors and tested using animal models. These results can only be transferred to humans in a limited sense. During the last year, we initiated a randomized study "Cryosurgery versus Conventional Surgery", whereby the peripheral blood and the normal skin from the areas surrounding the resection were compared. We were able to demonstrate in the peripheral blood of 8 cryosurgery patients a postoperative increase in the total and helper T-cells, HLA-DR-positive cells, and the ratio helper/suppressor T-cells in comparison to preoperative values. In the 8 patients treated with conventional surgery, these parameters decreased slightly or remained the same. The differences were highly significant (p = 0.001) to significant (p = 0.01). The results from the first 16 are patients studied presented and discussed here.

Topographical analysis of proliferation ([3H]thymidine labelling index and mitotic index) as compared with tumour growth and tumour weight in xenotransplanted melanoma. Changes due to local and systemic application of azelaic acid.

Xenotransplanted human melanoma was investigated by measuring the increase in tumour volume and in final tumour weight (macroscopical parameters) and histomorphological parameters of cell proliferation: Mitotic index (MI) and autoradiographic [3H]thymidine labelling index (LI). A total of 87 tumours, derived from a human melanoma metastasis and a primary nodular melanoma respectively, were analysed by these methods in two series. Topical treatment of the tumours with azelaic acid cream resulted in a statistically significant reduction in the increase in tumour volume and, in the first series, in a clear decrease in final tumour weight and in the MI, as compared with controls. The LI was decreased only in the superficial region of the tumours, i.e. at the site of treatment. Subtumoral injection of azelaic acid (disodium salt solution) was the second route of local therapy. It was followed by a significant reduction in the increase in tumour volume, of final tumour weight (first series) and in the MI. The average LI was clearly smaller than in the controls, especially at the tumour base, which was the site of injection (local effect). Systemic (intravenous) injection of azelaic acid (same concentration of the disodium salt solution) had no negative effect on the increase in tumour volume or final tumour weight, but was followed by a clear reduction of the MI. The average LI of this group was significantly smaller than in the controls as well. This effect was most impressive in the perivascular regions of large and small vessels, which fact can be interpreted as a sort of local effect via the blood stream after systemic application of azelaic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Melanoma in situ (MIS) adjacent to an invasive nodular melanoma ("SSM/NM") and its metastases--DNA-cytophotometry, mitotic index, and anisokaryosis.

Malignant melanomas of the superficial spreading type usually have an intraepidermal tumour component in their periphery which frequently displays the morphological features of a melanoma in situ (adjacent MIS). It is thus comparable to exclusively epidermal melanomas; melanoma in situ (MIS). Taking 10 superficial melanomas with a nodular component ("SSM/NM") 31 adjacent MIS regions and 36 nodular melanoma components were analysed in serial tissue slides. Planimetric estimation of the nuclear areas was employed as a measure of anisokaryosis. DNA-Feulgen-cytophotometry was applied to obtain an objective variable in judging malignancy in the DNA-histographs (paraffin material). Furthermore we investigated 8 metastases of one of the malignant melanomas applying the methods described. A comparison of the epidermal with the invasive tumour components revealed an increase in the nuclear area which, however, decrease from the superior to the inferior nodular regions and which are further reduced in melanoma metastases. Anisokaryosis is evidently less in metastases compared with all primary melanomas. The nuclear DNA-content increases from the epidermal to the invasive tumour compartments and is lower in the inferior nodular regions when compared with the superior ones. No further significant differences are, however, established in the metastases. The coefficients of variability of the DNA-contents, being a potential indicator of DNA-heterogeneity reflect higher values in the epidermal tumour components compared with the nodular regions, decreasing from the superior to the inferior nodular parts of the tumour. All metastases have smaller values than the respective primary melanoma. In the DNA-histographs 75% of the intraepidermal tumour components have obvious signs of malignancy including tumour cell stem lines in 19% of the cases. 85% of the nodular regions investigated have clear signs of malignancy, 33% of which also have aneuploid stem lines. All metastases have obvious signs of malignancy and tumour cell stem lines in 50% of the cases observed. The following conclusions can be drawn from our findings: DNA-Feulgen-cytophotometry and nuclear planimetry are additional feasible methods for judging the epidermal component of a melanocytic lesion as malignant (adjacent melanoma in situ) on paraffin material. Furthermore these methods give different results in invasive nodular versus epidermal (in situ) melanoma components. Both the DNA-histographs and our immunohistochemical investigations (monoclonal antibody P 3.58) indicate the malignant potential of adjacent MIS.(ABSTRACT TRUNCATED AT 400 WORDS)

[Skin changes in advanced age--biochemical findings corresponding to morphology?]. / Hautveränderungen im hohen Lebensalter--korrespondieren biochemische Befunde zur Morphologie?

The clinically relevant morphological changes of the skin during aging can be summarized by the term "senile atrophy". The changes are a diminished thickness of epidermis with a reduced mitosis rate of epidermal basal cells, shortened and attenuated rete ridges, reduction of epidermal appendages, and a decreased number of fibroblasts and capillaries in the dermis. Corresponding to these morphological findings regarding the cell number in the senile skin (cutis) we found a slight decrease in the DNA concentration of human and rat cutis. The specific DNA activity (3H-thymidine incorporation rate related to DNA concentration) decreased in presenile versus adult animals. The mesenchymal changes in the dermis have been morphologically described by the term "senile elastosis" or "elastoid collagen degeneration", but in fact they correspond to a progressive collagen denaturation with aging. The total collagen concentration, here determined as the hydroxyproline concentration in the human cutis, shows almost constant values from the 3rd until the 9th decade of life in both sexes. This is also true for the skin of two different rat strains. The insoluble collagen fraction shows a relative increase to the disadvantage of the soluble collagen fractions, which can be interpreted as an indicator of a decelerated collagen turnover. In spite of the decelerated turnover, i.e. a prolonged half-life of the collagen metabolism in the skin, the indicators of the collagen neosynthesis (14C-proline incorporation rate, specific hydroxyproline activity, prolyl-hydroxylase activity) are significantly elevated in the cutis of presenile versus adult rats. Any connection of these findings with a possible change in the distribution of collagen types in the senile skin (e.g. pericapillar fibrosis with increase of collagen type I as well as changes in the distribution of type I, III, IV and V) can only be discussed at present. The glycosaminoglycans in the cutis show a minimal increase of the total content of hexosamines and uronic acids with a significant shift in the ratio of the glycosaminoglycan components in favour of dermatan sulfate and keratan sulfate and to the disadvantage of hyaluronic acid and partly also of chondroitin-4-sulfate and -6-sulfate. The neosynthesis of sulfated glycosaminoglycans (indicator method: 35S-sulfate incorporation rate) is only slightly increased whereas the enzyme activities being specific for the glycosaminoglycan catabolism (beta-glucuronidase, beta-N-acetyl-glucosaminidase) are significantly decreased with aging of the skin.(ABSTRACT TRUNCATED AT 400 WORDS)

[Age-related changes in cell content (DNA) and glycosaminoglycan-degrading enzymes (beta-glucuronidase, beta-N-acetylglucosaminidase of connective tissue and parenchymal organs of the rat caused by 6-methylprednisolone (enzyme induction, adaptation, acceleration of maturation and possible modification of aging]. / Altersabhängige Veränderungen des Zellgehaltes (DNA) und der Glykosaminoglykan-abbauenden Enzyme (beta-Glucuronidase, beta-N-Acetylglucosaminidase) bindegewebiger und parenchymatöser Organe der Ratte durch 6-Methylprednisolon (Enzyminduktion, Adaptation, Reifungsbeschleunigung und mögliche Alternsbeeinflussung).

This paper is based on previous investigations, which had shown an evident acceleration of maturation and enzyme induction in several organs, not only in the lung, due to a pre- and postnatal application of prednisolone. Applying the same dosage we now investigated whether there is a similar effect of a short-term application of prednisolone in mesenchymal and parenchymal organs of young adult and presenile rats of the same strain (Chbb: THOM/SPF) analyzing the physiological cell regeneration (DNA concentration) as well as functional parameters of the glycosaminoglycan metabolism (e.g. the lysosomal enzymes beta-glucuronidase and beta-N-acetylglucosaminidase). The results show a significant age-dependent decrease of the DNA concentration (lung, spleen, skin, and rib cartilage), a significant age-dependent decrease of the total activity of the beta-glucuronidase (kidney, rib cartilage, and skin) or a significant age-dependent increase of this enzyme activity (spleen and liver) respectively as well as a significant decrease of the beta-N-acetylglucosaminidase activity (skin and rib cartilage) or a significant increase of this enzyme activity (spleen and lung). After application of prednisolone the rats showed a significant reduction of the DNA concentration only in the skin of young adult rats, but no changes in the other organs of the young adult or presenile animals compared to untreated controls. Similar to our findings after postnatal prednisolone application, we found the greatest increases or decreases respectively of the activities of these lysosomal enzymes due to 2- to 3-fold or 4- to 5-fold prednisolone application. Again similar to our previous findings, we found the phenomena of adaptation and rebound effects including the so-called over-compensation in the young adult and especially in the presenile rats but these effects were delayed and weaker in most of the older animals compared to the young adult rats.