Explant culture of rat colon: A model system for studying metabolism of chemical carcinogens

Summary An explant culture system has been developed for the long-term maintenance of colonic tissue from the rat. Explants of 1 cm² in size were placed in tissue-culture dishes to which was added 2 ml of CMRL-1066 medium supplemented with glucose, hydrocortisone, β-retinyl acetate, and either 2.5% bovine albumin or 5% fetal bovine serum. The dishes were placed in a controlied-atmosphere chamber which was gassed with 95% O₂ and 5% CO₂. The chamber then was placed on a rocker platform which rocked at 10 cycles per min causing the medium to flow intermittently over the epithelial surface. The explants were incubated at 30°C. The viability of the tissue was measured both by incorporation of specific precursors into cellular macromolecules and by monitoring of tissue morphology with light and electron microscopy. Cultured rat colon was able to metabolize benzo[a]pyrene, 7,12-dimethylbenz[a]anthracene, aflatoxin B₁, dimethylnitrosamine, 1,2-dimethylhydrazine, and methylazoxymethanol acetate into chemical species that bind to cellular DNA and protein.     

Superinfection exclusion by bacteriophage T7.

Only two of the early genes of bacteriophage T7 were found to play a significant role in exclusion of superinfecting bacteriophage T3 particles; genes 0.3 and 1. Protein synthesis by the preinfecting phage particle was not required for efficient exclusion. These findings are discussed with regard to the known functions of these genes during T7 development.     

Reconciling emergences: An information-theoretic approach to identify causal emergence in multivariate data

The broad concept of emergence is instrumental in various of the most challenging open scientific questions—yet, few quantitative theories of what constitutes emergent phenomena have been proposed. This article introduces a formal theory of causal emergence in multivariate systems, which studies the relationship between the dynamics of parts of a system and macroscopic features of interest. Our theory provides a quantitative definition of downward causation, and introduces a complementary modality of emergent behaviour—which we refer to as causal decoupling. Moreover, the theory allows practical criteria that can be efficiently calculated in large systems, making our framework applicable in a range of scenarios of practical interest. We illustrate our findings in a number of case studies, including Conway’s Game of Life, Reynolds’ flocking model, and neural activity as measured by electrocorticography.     

Assessing changes in subjective and objective function from pre- to post-knee arthroplasty using the Cardiff Dempster–Shafer theory classifier

The purpose of this study is to assess changes in subjective and objective function from pre- to post-knee arthroplasty (KA) using a combined classifier technique. Twenty healthy adults (50–80 years) and 31 KA patients (39–81 years) were studied (4 weeks pre- and 6 months post-KA). Questionnaire measures of subjective pain, joint stability, activity and function were collected. Objective functional assessment included goniometry, ultrasound imaging and 3-D motion analysis/inverse modelling of gait and sit–stand. An optimal set of variables were used to classify function using the Cardiff Dempster–Shafer theory (DST) method. Out of sample accuracy of the classifiers ranged between 90% and 94% for segregating healthy individuals and pre-KA patients. Post-KA subjective function improved with 74% classified as healthy. However, there was minimal improvement in objective measures (23% classified as healthy). The novel use of Cardiff DST segregated KA patients from healthy individuals and estimated changes in function from pre- to post-surgery. KA patients had improved pain and function post-operation but objective knee joint measures remained different to healthy individuals.     

Economic considerations for the eradication endgame

An infectious disease will be eradicated only if it is eliminated everywhere, including in the hardest-to-reach, most vaccine-wary communities. If eradication is successful, it promises a dividend in the form of avoided infections and vaccinations. However, success is never certain unless and until eradication is achieved, and claiming the dividend means bearing the possibly great risk of re-emergence. Economic analysis of eradication evaluates these risks and rewards relative to the alternative of ‘optimal control’, and also exposes the incentives for achieving and capitalizing on eradication. Eradication is a ‘game’, because some countries may be willing to eliminate the disease within their borders only if assured that all others will eliminate the disease within their borders. International financing is also a game, because each country would rather free ride than contribute. Finally, for diseases such as polio, capitalizing on eradication is a game, for should any country continue to vaccinate in the post-eradication era using the live-attenuated polio vaccine, the countries that stop vaccinating will be exposed to the risk of vaccine-derived polioviruses. In the framework developed in this paper, eradication is a seductive goal, its attainment fraught with peril.     

Within-litter variance in rat maternal behaviour

As a first step in determining the influence of maternal behaviour on sibling behavioural variance, we tested whether rat mothers differentially interact with neonates within the same litter. We also tested whether fading of an ink-mark on individual pups could provide an index of within-litter variance in maternal licking in laboratory rats. In Study 1, during the first postnatal week we distinguished individual Sprague-Dawley rat pups across 4 litters by placing an ink-mark on the skin and quantified variance in maternal licking frequency toward each pup and compared fading of individual pup marks to the frequency of maternal licks received and to four pup characteristics that could influence mark-fading. In Study 2, neonate mark-fading (a proxy for maternal licking) was compared to adolescent and adult offspring behaviour across 8 litters. Results indicated that: (1) there are substantial and consistent differences in how much rat mothers lick same-sex siblings within a litter, (2) differential licking rates can be documented with a non-observational method (ink-mark-fading), and (3) within-litter variance in maternal behaviour may relate to sibling behavioural variance. The findings indicate a viable research model for future experimental studies on causes and consequences of differential maternal investment within families.     

Host Reticulocytes Provide Metabolic Reservoirs That Can Be Exploited by Malaria Parasites

Human malaria parasites proliferate in different erythroid cell types during infection. Whilst Plasmodium vivax exhibits a strong preference for immature reticulocytes, the more pathogenic P. falciparum primarily infects mature erythrocytes. In order to assess if these two cell types offer different growth conditions and relate them to parasite preference, we compared the metabolomes of human and rodent reticulocytes with those of their mature erythrocyte counterparts. Reticulocytes were found to have a more complex, enriched metabolic profile than mature erythrocytes and a higher level of metabolic overlap between reticulocyte resident parasite stages and their host cell. This redundancy was assessed by generating a panel of mutants of the rodent malaria parasite P. berghei with defects in intermediary carbon metabolism (ICM) and pyrimidine biosynthesis known to be important for P. falciparum growth and survival in vitro in mature erythrocytes. P. berghei ICM mutants (pbpepc^-, phosphoenolpyruvate carboxylase and pbmdh^-, malate dehydrogenase) multiplied in reticulocytes and committed to sexual development like wild type parasites. However, P. berghei pyrimidine biosynthesis mutants (pboprt^-, orotate phosphoribosyltransferase and pbompdc^-, orotidine 5′-monophosphate decarboxylase) were restricted to growth in the youngest forms of reticulocytes and had a severe slow growth phenotype in part resulting from reduced merozoite production. The pbpepc^-, pboprt^- and pbompdc^- mutants retained virulence in mice implying that malaria parasites can partially salvage pyrimidines but failed to complete differentiation to various stages in mosquitoes. These findings suggest that species-specific differences in Plasmodium host cell tropism result in marked differences in the necessity for parasite intrinsic metabolism. These data have implications for drug design when targeting mature erythrocyte or reticulocyte resident parasites.