Thermal and mechanical effects of different excitation modes based on low frequency laser modulation in optical hyperthermia

The low frequency modulation of the laser source (menor que30KHz) allows the generation of a pulsed signal that intermittently excites the gold nanorods. The temperature curves obtained for different frequencies and duty cycles of modulation but with equal average power and identical laser parameters, show that the thermal behavior in continuous wave and modulation modes is the same. However, the cell death experiments suggest that the percentage of death is higher in the cases of modulation. This observation allows us to conclude that there are other effects in addition to temperature that contribute to the cellular death. The mechanical effects like sound or pressure waves are expected to be generated from thermal expansion of gold nanorods. In order to study the behavior and magnitude of these processes we have developed a measure device based on ultrasound piezoelectric receivers (25KHz) and a lock-in amplifier that is able to detect the sound waves generated in samples of gold nanorods during laser irradiation providing us a voltage result proportional to the pressure signal. The first results show that the pressure measurements are directly proportional to the concentration of gold nanorods and the laser power, therefore, our present work is focused on determine the real influence of these effects in the cell death process.

Nucleotide excision repair in rat male germ cells: low level of repair in intact cells contrasts with high dual incision activity in vitro

The acquisition of genotoxin-induced mutations in the mammalian germline is detrimental to the stable transfer of genomic information. In somatic cells, nucleotide excision repair (NER) is a major pathway to counteract the mutagenic effects of DNA damage. Two NER subpathways have been identified, global genome repair (GGR) and transcription-coupled repair (TCR). In contrast to somatic cells, little is known regarding the expression of these pathways in germ cells. To address this basic question, we have studied NER in rat spermatogenic cells in crude cell suspension, in enriched cell stages and within seminiferous tubules after exposure to UV or N-acetoxy-2-acetylaminofluorene. Surprisingly, repair in spermatogenic cells was inefficient in the genome overall and in transcriptionally active genes indicating non-functional GGR and TCR. In contrast, extracts from early/mid pachytene cells displayed dual incision activity in vitro as high as extracts from somatic cells, demonstrating that the proteins involved in incision are present and functional in premeiotic cells. However, incision activities of extracts from diplotene cells and round spermatids were low, indicating a stage-dependent expression of incision activity. We hypothesize that sequestering of NER proteins by mispaired regions in DNA involved in synapsis and recombination may underlie the lack of NER activity in premeiotic cells.