Newts, such as Notophthalmus viridescens, can regenerate many structures after amputation or injury and have also shown a refractory response to the formation of cancer in tissues that have regenerative capabilities. The mechanisms behind this latter ability have surprisingly not been studied. In the current study, N. viridescens were exposed to a variety of carcinogens in tissue that cannot regenerate with the intention of inducing tumour formation. After testing multiple carcinogens, multiple sites of injection, and two different modes of delivery, no tumours were generated. Consequently, in vitro assays were developed in order to better understand this ability of newt cells to evade transformation. Mouse and newt muscle cells were exposed to DNA damaging agents, such as irradiation and carcinogens, in culture and their response was monitored with respect to the DNA damage response proteins γ-H2AX, p53, and phospho-p53. These proteins are important as they help prevent mutations in the genome from being passed on to daughter cells and potentially generating cells that proliferate uncontrollably, a hallmark of cancer. Preliminary results suggest that after irradiation, γ-H2AX is present in newt cells for a considerably longer period of time in comparison to mouse cells. p53, as well as phospho-p53, appear to be present at a basal level before and after irradiation in newt cells, whereas mouse cells have a distinct increase upon damage and decrease upon repair. The carcinogen treatments also suggest that newt cells have basal levels of expression of these proteins prior to treatment. These studies suggest that newt cells may have a unique profile of these DNA damage response proteins and may be “primed” to repair any future damage. This is a good first step in understanding what is likely a very complicated explanation for newts’ refractory response to cancer formation.

Newts, such as Notophthalmus viridescens, can regenerate many structures after amputation or injury and have also shown a refractory response to the formation of cancer in tissues that have regenerative capabilities. The mechanisms behind this latter ability have surprisingly not been studied. In the current study, N. viridescens were exposed to a variety of carcinogens in tissue that cannot regenerate with the intention of inducing tumour formation. After testing multiple carcinogens, multiple sites of injection, and two different modes of delivery, no tumours were generated. Consequently, in vitro assays were developed in order to better understand this ability of newt cells to evade transformation. Mouse and newt muscle cells were exposed to DNA damaging agents, such as irradiation and carcinogens, in culture and their response was monitored with respect to the DNA damage response proteins γ-H2AX, p53, and phospho-p53. These proteins are important as they help prevent mutations in the genome from being passed on to daughter cells and potentially generating cells that proliferate uncontrollably, a hallmark of cancer. Preliminary results suggest that after irradiation, γ-H2AX is present in newt cells for a considerably longer period of time in comparison to mouse cells. p53, as well as phospho-p53, appear to be present at a basal level before and after irradiation in newt cells, whereas mouse cells have a distinct increase upon damage and decrease upon repair. The carcinogen treatments also suggest that newt cells have basal levels of expression of these proteins prior to treatment. These studies suggest that newt cells may have a unique profile of these DNA damage response proteins and may be “primed” to repair any future damage. This is a good first step in understanding what is likely a very complicated explanation for newts’ refractory response to cancer formation.

Newts, such as Notophthalmus viridescens, can regenerate many structures after amputation or injury and have also shown a refractory response to the formation of cancer in tissues that have regenerative capabilities. The mechanisms behind this latter ability have surprisingly not been studied. In the current study, N. viridescens were exposed to a variety of carcinogens in tissue that cannot regenerate with the intention of inducing tumour formation. After testing multiple carcinogens, multiple sites of injection, and two different modes of delivery, no tumours were generated. Consequently, in vitro assays were developed in order to better understand this ability of newt cells to evade transformation. Mouse and newt muscle cells were exposed to DNA damaging agents, such as irradiation and carcinogens, in culture and their response was monitored with respect to the DNA damage response proteins γ-H2AX, p53, and phospho-p53. These proteins are important as they help prevent mutations in the genome from being passed on to daughter cells and potentially generating cells that proliferate uncontrollably, a hallmark of cancer. Preliminary results suggest that after irradiation, γ-H2AX is present in newt cells for a considerably longer period of time in comparison to mouse cells. p53, as well as phospho-p53, appear to be present at a basal level before and after irradiation in newt cells, whereas mouse cells have a distinct increase upon damage and decrease upon repair. The carcinogen treatments also suggest that newt cells have basal levels of expression of these proteins prior to treatment. These studies suggest that newt cells may have a unique profile of these DNA damage response proteins and may be “primed” to repair any future damage. This is a good first step in understanding what is likely a very complicated explanation for newts’ refractory response to cancer formation.

Newts, such as Notophthalmus viridescens, can regenerate many structures after amputation or injury and have also shown a refractory response to the formation of cancer in tissues that have regenerative capabilities. The mechanisms behind this latter ability have surprisingly not been studied. In the current study, N. viridescens were exposed to a variety of carcinogens in tissue that cannot regenerate with the intention of inducing tumour formation. After testing multiple carcinogens, multiple sites of injection, and two different modes of delivery, no tumours were generated. Consequently, in vitro assays were developed in order to better understand this ability of newt cells to evade transformation. Mouse and newt muscle cells were exposed to DNA damaging agents, such as irradiation and carcinogens, in culture and their response was monitored with respect to the DNA damage response proteins γ-H2AX, p53, and phospho-p53. These proteins are important as they help prevent mutations in the genome from being passed on to daughter cells and potentially generating cells that proliferate uncontrollably, a hallmark of cancer. Preliminary results suggest that after irradiation, γ-H2AX is present in newt cells for a considerably longer period of time in comparison to mouse cells. p53, as well as phospho-p53, appear to be present at a basal level before and after irradiation in newt cells, whereas mouse cells have a distinct increase upon damage and decrease upon repair. The carcinogen treatments also suggest that newt cells have basal levels of expression of these proteins prior to treatment. These studies suggest that newt cells may have a unique profile of these DNA damage response proteins and may be “primed” to repair any future damage. This is a good first step in understanding what is likely a very complicated explanation for newts’ refractory response to cancer formation.

No description available.

No description available.

2,5-Diphenyloxazole (PPO) was tested as a potential alternate inducer for the aryl hydrocarbon hydroxylase (AHH) system. Its apparent lack. of carcinogenicity and toxicity provide a possible system for investigation of enzyme systems related to chemical carcinogenesis without exposure of the researcher to potent carcinogenic compounds. These studies found PPO to be an inducer of AHH in cultured human lymphocytes. When PPO was utilized as a substrate for the AHH assay system, the major metabolites produced were strongly fluorescent. A simple fluorometric assay was developed which employed PPO as the substrate and which measured constitutive activity more efficiently than similar assays using benzo(a)pyrene as the substrate. Quantitation of both basal and induced lymphocyte AHH metabolism of PPO may be applicable to human population studies and may provide a tool to determine possible genetic variables with respect to carcinogen metabolism related to cancer risk.

No description available.

Würzburg, University, Diss., 2007.

Recent European legislation has enforced a reduction in the use of animal models for safety assessment purposes and carcinogenicity testing. The Syrian hamster embryo cell transformation assay (SHE CTA) has been proposed as a suitable animal alternative, but its implementation into test batteries has been delayed. This is due to concerns regarding the assay’s endpoint subjectivity and, moreover, the model’s relevance to carcinogenicity remains mostly unexplored. Senescence is an essential barrier against uncontrolled cell proliferation and its evasion is necessary for clonal evolution and tumour development. Carcinogenesis can be modelled by reproducing underlying mechanisms leading to senescence bypass. In this project, the SHE CTA was performed using the known mutagen and human carcinogen, benzo(a)pyrene, and the resulting SHE colonies were analysed. It was found that morphological transformation (MT) does not guarantee senescence bypass and cell immortalisation, but increases the likelihood of MT-derived cells subsequently acquiring unlimited growth potential. A limited number (between 10 and 20 %) of MT colonies produced cell clones capable of sustained proliferation and in most cases secondary events were necessary for the evasion of senescence barriers. With regard to mechanisms, p53 point mutations were present in 30 % of immortal B(a)P-induced MT colony-derived cells and located within the protein’s DNA binding domain. No p16 mutations were identified. Expression of p16 mRNA was commonly silenced or markedly reduced by a combination of mechanisms including monoallelic deletion, promoter methylation and BMI-1 overexpression. Taking advantage of the recently available Syrian hamster genomic sequence information generated by the Broad Institute, the coding regions of the Syrian hamster CDKN2A/B locus were shown to have good homology to human nucleotide sequences and confirmed the exonic structures of SH p16, ARF and p15. The findings further implicate the importance of p16 in regulating senescence while providing a molecular evaluation of SHE CTA-derived MT clones.