Ultraviolet B-induced melanoma in Monodelphis domestica occurs in the absence of alterations in the structure or expression of the p53 gene.

Monodelphis domestica, a South American opossum, has been established as a mammalian model for sporadic ultraviolet radiation (UVR)-induced melanoma. Using this model system, we investigated the role of changes in the p53 gene in the development of cutaneous melanocyte-derived lesions. Cutaneous melanocytic hyperplasias, benign melanomas and metastatic primary melanomas, plus affected lymph nodes and visceral organs, were screened for mutations in the Monodelphis p53 gene by single-strand conformation polymorphism analysis and direct sequencing. With the exception of a silent point mutation found in a single benign melanocytic hyperplasia sample, no p53 mutations were detected. Furthermore, a relative quantitative reverse transcriptase-polymerase chain reaction approach was used to analyse p53 gene expression at different stages of primary melanoma progression and revealed no substantial changes in p53 mRNA levels. These results suggest that, as in humans, UVR-induced melanoma in the Monodelphis model is initiated and progresses on the basis of predominantly p53-independent molecular pathways.

Characterization of the CDKN2A and ARF genes in UV-induced melanocytic hyperplasias and melanomas of an opossum (Monodelphis domestica).

We examined the involvement of the cyclin-dependent kinase inhibitor 2A (CDKN2A) locus in the pathogenesis of ultraviolet (UV) radiation-induced melanomas in an opossum (Monodelphis domestica) melanoma model in which suckling young were exposed to UVB to produce melanocytic lesions. Monodelphis CDKN2A and alternated reading frame (ARF) cDNAs were cloned and sequenced, and the expression patterns of these genes were determined by reverse transcription-polymerase chain reaction in normal tissues, 39 primary melanocytic skin lesions, and two tumor-derived cell lines, one nonmetastatic and one metastatic. Primary melanocytic lesions, including hyperplasias, benign melanomas, melanomas metastatic to lymph nodes, and melanomas metastatic to nodes and additional visceral organs, were categorized accordingly as types I-IV. Levels of CDKN2A transcripts were most abundant in type III tumor samples and the metastatic cell line but absent in the nonmetastatic cell line. ARF transcripts were expressed in all tumors and cell lines. A UV-signature mutation was detected with the wild-type allele at the CDKN2A locus in type II and III primary tumor samples and in the nonmetastatic cell line. Interestingly, in the metastatic cell line, only the mutant allele was present and expressed. These data suggest dynamic changes in the expression and/or structure of the CDKN2A and ARF genes represent one molecular defect associated with the etiology of melanoma formation and progression in the Monodelphis model system.

Bubble chamber as a trace chemical detector.

A novel concept for trace chemical analysis in liquids has been demonstrated. The technique utilizes light absorption in a superheated liquid. Although a superheated liquid is thermodynamically unstable, a high degree of superheating can be dynamically achieved for a short period of time. During this time the superheated liquid is extremely sensitive to boiling at nucleation sites produced by energy deposition. Observation of bubbles in the superheated liquid in some sense provides amplification of the initial energy deposition. Bubble chambers containing superheated liquids have been used to detect energetic particles; now a bubble chamber is used to detect a trace chemical in superheated liquid propane by observing bubble formation initiated by optical absorption. Crystal violet is used as a test case and can be detected at the subpart-per-10(12) level by using a Nd:YAG laser. The mechanism for bubble formation and ideas for further improvement are discussed.