Multi-objective Bayesian algorithm automatically discovers low-cost high-growth serum-free media for cellular agriculture application

In this work, we applied a multi-information source modeling technique to solve a multi-objective Bayesian optimization problem involving the simultaneous minimization of cost and maximization of growth for serum-free C2C12 cells using a hyper-volume improvement acquisition function. In sequential batches of custom media experiments designed using our Bayesian criteria, collected using multiple assays targeting different cellular growth dynamics, the algorithm learned to identify the trade-off relationship between long-term growth and cost. We were able to identify several media with $>100\%$ more growth of C2C12 cells than the control, as well as a medium with 23% more growth at only 62.5% of the cost of the control. These algorithmically generated media also maintained growth far past the study period, indicating the modeling approach approximates the cell growth well from an extremely limited data set.     

Soil arthropods as indicators of water stress in Antarctic terrestrial habitats?

Abiotic features of Antarctic terrestrial habitats, particularly low temperatures and limited availability of liquid water, strongly influence the ecophysiology and life histories of resident biota. However, while temperature regimes of a range of land microhabitats are reasonably well characterized, much less is known of patterns of soil water stress, as current technology does not allow measurement at the required scale. An alternative approach is to use the water status of individual organisms as a proxy for habitat water status and to sample over several years from a population to identify seasonal or long‐term patterns. This broad generalization for terrestrial invertebrates was tested on arthropods in the maritime Antarctic. We present analyses of a long‐term data set of body water content generated by monthly sampling for 8–11 years of seven species of soil arthropods (four species of Acari, two Collembola and one Diptera) on maritime Antarctic Signy Island, South Orkney Islands. In all species, there was considerable within‐ and between‐sample variability. Despite this, clear seasonal patterns were present in five species, particularly the two collembolans and a prostigmatid mite. Analyses of monthly water content trends across the entire study period identified several statistically significant trends of either increase or decrease in body water content, which we interpret in the context of regional climate change. The data further support the separation of the species into two groups as follows: firstly, the soft‐bodied Collembola and Prostigmata, with limited cuticular sclerotization, which are sensitive to changes in soil moisture and are potentially rapid sensors of microhabitat water status, secondly, more heavily sclerotized forms such as Cryptostigmata (=Oribatida) and Mesostigmata mites, which are much less sensitive and responsive to short‐term fluctuations in soil water availability. The significance of these findings is discussed and it is concluded that annual cycles of water content were driven by temperature, mediated via radiation and precipitation, and constituted reliable indicators of habitat moisture regimes. However, detailed ecophysiological studies are required on particular species before such information can be used to predict over long timescales.     

In-situ enzyme histochemistry on plastic-embedded plant material. The development of an artefact-free β-glucuronidase assay

An in-situ enzyme histochemical method is described that preserves the tissue and cell structure as well as the enzyme activities. Flower buds at different developmental stages from wild-type and transgenic Brassica napus plants, the latter containing the GUS gene under the control of a tapetum-specific promoter, were used as starting material. The method is based on the following principles: processing of the tissue on crushed ice, no fixation but a pretreatment with spermidine, partial dehydration with acetone, and a final embedding in a water-miscible glycol methacrylate resin at 5°C. This method was used to set up a sensitive p-glucuronidase histochemical assay with a high resolution. A succinate dehydrogenase assay was included as a control for tissue and cell viability as well as a standard for the relative metabolic activities between different tissues and cell types.     

Paraquat-induced DNA damage in mammalian cells

We have examined the possibility that paraquat (PQ) may exert its toxicity by inducing DNA damage. Mouse lymphoblasts in culture exhibited inhibition of colony forming ability and DNA single strand breaks following a 2 hour exposure to PQ. These phenomenon are dose dependent and increase when a rat liver S9 fraction is included in the incubation mixture. The presence of superoxide dismutase and catalase did not prevent the effects of PQ. Our data indicate that DNA should be considered as a possibile macromolecular target for the lethal effects of paraquat.     

The Effects of Lithium on a Neuronal in Vitro Orcadian Pacemaker

Previous studies have suggested a causal connection between abnormalities of the circadian system and affective disorders. The effectiveness of lithium or rubidium as a treatment for affective disorders and the ability of lithium or rubidium to influence circadian pacemakers has stimulated research into the mechanism of lithium's action on circadian systems. In this study we used a neuronal in vitro circadian pacemaker preparation, the eye of the mollusc Bulla, to examine the cellular effects of lithium and rubidium. Continuous extracellular LiCl application lengthens the period of the circadian rhythm of the Bulla pacemaker in a concentration-dependent manner. Rubidium was found to be more effective than lithium in period lengthening. Stable phase delays were generated by 2-h pulses of 395 mM LiCl applied extracellularly from zeitgeber time (ZT) 5-7 (mid subjective day). Concomitant continuous application of 16 mM LiCl and light (a depolarizing agent) generated period lengthening substantially greater than the arithmetic sum of the modest period lengthening of each treatment alone. Furthermore, LiCl pulses, applied together with depolarizing extracellular KC1 concentrations, yielded an increasing magnitude of phase delays with increasing KC1 concentration. These data suggest that LiCl acts intracellularly on the circadian pacemaker cells by entering through a voltage-dependent channel, most likely a sodium channel.