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Biomedical subjects

A Lerche

Publications and source records attributed to A Lerche.

4 recordsLinked to original sources

DNA in psoriatic epidermis.

An electron microscopic technique has been used to visualize crosslinks after total denaturation on DNA isolated from epidermis and dermis in patients with psoriasis treated with 8-methoxypsoralen (8-MOP) and irradiation with ultraviolet light at 360 nm (PUVA treatment). This technique facilitated accurate measurements of the number and density of DNA interstrand crosslinks. 30 biopsies from 14 patients were studied and a total of 9503 DNA molecules were scored in the electron-microscope, 6 patients were treated topically with 8-MOP and 10 were on systemic treatment. Two of the patients on topical treatment had previously been on systemic treatment. 1% of all DNA molecules contained 3 or more cross-links. The overall frequency of cross-links was almost identical in the epidermis (1.1%) and in the dermis (0.9%) and, furthermore, virtually the same in patients on topical and systemic PUVA treatment. The total number of crosslinks was of the same magnitude as that previously found in normal human skin. (V. Bohr et al., Acta Dermatovener, in press.) No significantly increased damage of the genetic material (DNA) was demonstrated in our study as a consequence of the PUVA treatment using 8-MOP. We have previously shown (V. Bohr & A. Lerche, Biochim Biophys Acta, in press) that 8-MOP induced crosslinks after irradiation at 360 nm in an in vitro system of pure DNA. In this system a correlation was established between the density of crosslinks formed and irradiation time, concentration of 8-MOP, and irradiation intensity.

DNA

DNA interstrand crosslinks visualized by electron microscopy in PUVA-treated psoriasis.

An electron microscopic method has been used to visualize crosslinks after total denaturation of DNA isolated from epidermis and dermis in patients with psoriasis treated with 8-methoxypsoralen (8-MOP) and irradiation with ultraviolet light at 360 nm (PUVA treatment). This technique enabled accurate measurement of the number and density of DNA interstrand crosslinks. 30 biopsies were studied and a total of 9503 DNA molecules were scored in the electron microscope. 6 patients were treated topically with 8-MOP and 10 were on systemic treatment and biopsies were obtained immediately after irradiation. 1% of the total DNA molecules examined contained 3 or more crosslinks. The overall frequency of crosslinks was almost identical in the epidermis (1.1%) and in the dermis (0.9%) and, furthermore, virtually the same in patients on topical and systemic PUVA treatment. The total number of crosslinks was of the same magnitude as those previously found by us in normal human skin. Thus, though the formation of interstrand DNA crosslinks can be considered as an indicator of damage of the genetic material, we were unable to demonstrate this as being a consequence of PUVA treatment using 8-MOP.

Adult

In vitro crosslinking of DNA by 8-methoxypsoralen visualized by electron microscopy.

By electron microscopic visualisation of totally denatured DNA, we have detected photochemically induced 8-methoxypsoralen crosslinks in vitro after irradiation at 360 nm. The amount of crosslinks was expressed as the percentage of DNA length which was kept in double-stranded appearance by closely situated crosslinks. This percentage correlated well with irradiation time, irradiation intensity, and the concentration of 8-methoxypsoralen. These parameters have also been correlated with the mean size and the size distribution of non-crosslinked regions of DNA, so called bubbles. For a comparison with another psoralen type, we have carried out a similar set of experiments using 4,5,8-trimethylpsoralen.

Coumarins

DNA interstrand crosslinks in normal human skin visualized by electron microscopy.

Normal human skin was investigated for DNA interstrand crosslinks by a denaturation electron microscopic method. Skin biopsy materials from the thighs of 8 healthy volunteers were separated into epidermis and dermis. DNA from each layer was isolated, purified and studied in the electron microscope. The presence of DNA interstrand crosslinks in normal human skin was demonstrated. The number of crosslinked DNA molecules and the density of crosslinking within the molecules were determined. Crosslinks were found in 0.8% of the scored molecules and were located almost exclusively in epidermal DNA. The intramolecular density of crosslinking showed a wide range of variation in each volunteer and between the volunteers.

Adult