NADPH:cytochrome c (P450) reductase activates tirapazamine (SR4233) to restore hypoxic and oxic cytotoxicity in an aerobic resistant derivative of the A549 lung cancer cell line

Tirapazamine (TPZ, SR4233, WIN 59075) is a bioreductive drug that is activated in regions of low oxygen tension to a cytotoxic radical intermediate. This labile metabolite shows high selective toxicity towards hypoxic cells, such as those found in solid tumours. Under aerobic conditions, redox cycling occurs with subsequent generation of superoxide radicals, which are also cytotoxic. NADPH:cytochrome c (P450) reductase (P450R) is a one-electron reducing enzyme that efficiently activates TPZ. Recently a derivative of the A549 non-small cell lung cancer cell line (A549c50) was generated that showed substantially reduced P450R activity compared to its parental line (Elwell et al (1997) Biochem Pharmacol54: 249–257). Here, it is demonstrated that the A549c50 cells are markedly more resistant to TPZ under both aerobic and hypoxic conditions. In addition, these cells have a dramatically impaired ability to metabolize TPZ to its two-electron reduction product, SR4317, under hypoxic conditions when compared to wild-type cells. P450R activity in the A549c50 cells was reintroduced to similar levels as that seen in the parental A549 cells by transfection of the full-length cDNA for human P450R. These P450R over-expressing cells exhibit restored sensitivity to TPZ under both aerobic and hypoxic conditions, comparable to that found in the original parental A549 cells. Further, the ability of the transfected cells to metabolize TPZ to SR4317 under hypoxic conditions is also shown to be restored. This provides further evidence that P450R can play an important role in the activation, metabolism and toxicity of this lead bioreductive drug. © 2000 Cancer Research Campaig

Electrically induced bacterial membrane-potential dynamics correspond to cellular proliferation capacity

Membrane-potential dynamics mediate bacterial electrical signaling at both intra- and intercellular levels. Membrane potential is also central to cellular proliferation. It is unclear whether the cellular response to external electrical stimuli is influenced by the cellular proliferative capacity. A new strategy enabling electrical stimulation of bacteria with simultaneous monitoring of single-cell membrane- potential dynamics would allow bridging this knowledge gap and further extend electrophysiological studies into the field of microbi- ology. Here we report that an identical electrical stimulus can cause opposite polarization dynamics depending on cellular proliferation capacity. This was demonstrated using two model organisms, namely Bacillus subtilis and Escherichia coli, and by developing an apparatus enabling exogenous electrical stimulation and single-cell time-lapse microscopy. Using this bespoke apparatus, we show that a 2.5-sec- ond electrical stimulation causes hyperpolarization in unperturbed cells. Measurements of intracellular K+ and the deletion of the K+ channel suggested that the hyperpolarization response is caused by the K+ efflux through the channel. When cells are preexposed to 400 ± 8 nm wavelength light, the same electrical stimulation depolarizes cells instead of causing hyperpolarization. A mathematical model extended from the FitzHugh–Nagumo neuron model suggested that the opposite response dynamics are due to the shift in resting mem- brane potential. As predicted by the model, electrical stimulation only induced depolarization when cells are treated with antibiotics, protonophore, or alcohol. Therefore, electrically induced membrane- potential dynamics offer a reliable approach for rapid detection of proliferative bacteria and determination of their sensitivity to anti- microbial agents at the single-cell level

Ecology and conservation of avian insectivores of the rainforest understory: A pantropical perspective

Avian insectivores of the tropical rainforest understory ("understory insectivores") are common, diverse, and often sensitive to disturbance of tropical forest, making them useful as sentinels of rainforest ecosystem change. At the 2013 joint American Ornithologists' Union and Cooper Ornithological Society meeting in Chicago, USA, researchers convened a symposium to address the ecology and conservation of understory insectivores. This Special Issue of Biological Conservation is the result of that symposium: a collection of articles that unites our efforts to further understand and conserve understory insectivores. In this introductory paper, we review the diversity and ecology of understory insectivores, identify threats to the guild, discuss hypotheses on drivers of population declines, and make suggestions for future research. Deforestation and forest degradation are the immediate threats to this guild, with agricultural expansion (particularly oil palm plantations), urbanization, road expansion and logging leading the list. Although vulnerabilities of this guild are most evident in the Neotropics, there are few studies from Asia and fewer still from Africa-we recommend increased geographic coverage. If we are to understand the vulnerabilities of understory insectivores from a pantropical perspective, researchers should prioritize understanding the most serious threats (e.g., edge effects, deforestation, fragmentation, etc.) and standardize efforts to gauge understory insectivores' response to these threats (e.g., via species richness, abundance, demographic metrics). A coordinated approach by researchers working in tropical rainforests across the globe can help us understand the ecology of understory insectivores and meaningfully apply conservation and management actions. © 2015

Evidence for habitual climbing in a Pleistocene hominin in South Africa

Bipedalism is a defining trait of the hominin lineage, associated with a transition from a more arboreal to a more terrestrial environment. While there is debate about when modern human-like bipedalism first appeared in hominins, all known South African hominins show morphological adaptations to bipedalism, suggesting that this was their predominant mode of locomotion. Here we present evidence that hominins preserved in the Sterkfontein Caves practiced two different locomotor repertoires. The trabecular structure of a proximal femur (StW 522) attributed to Australopithecus africanus exhibits a modern human-like bipedal locomotor pattern, while that of a geologically younger specimen (StW 311) attributed to either Homo sp. or Paranthropus robustus exhibits a pattern more similar to nonhuman apes, potentially suggesting regular bouts of both climbing and terrestrial bipedalism. Our results demonstrate distinct morphological differences, linked to behavioral differences between Australopithecus and later hominins in South Africa and contribute to the increasing evidence of locomotor diversity within the hominin clade