A human breast tumor cell line (BT-474) that supports mouse mammary tumor virus replication

Summary A human breast tumor cell line BT-474 derived from an invasive ductal carcinoma was experimentally infected in vitro with a mouse mammary tumor virus from the RIII strain (RIII-MuMTV). The virus that replicated in the human cells was characterized as a mouse virus by immunofluorescence, electron microscopy and the presence of a specific RNA-directed DNA polymerase. The cells themselves were human as per the karyotype and isoenzyme migration patterns. It is concluded that human cells are susceptible to the mouse mammary tumor virus and can, eventually, support its replication. This work was supported by USPHS Grant CA-08515 from the National Cancer Institute and by NIH Contract N01-CP-81003.     

Pressure Cycling Technology Assisted Mass Spectrometric Quantification of Gingival Tissue Reveals Proteome Dynamics during the Initiation and Progression of Inflammatory Periodontal Disease

Understanding the progression of periodontal tissue destruction is at the forefront of periodontal research. The authors aimed to capture the dynamics of gingival tissue proteome during the initiation and progression of experimental (ligature‐induced) periodontitis in mice. Pressure cycling technology (PCT), a recently developed platform that uses ultra‐high pressure to disrupt tissues, is utilized to achieve efficient and reproducible protein extraction from ultra‐small amounts of gingival tissues in combination with liquid chromatography‐tandem mass spectrometry (MS). The MS data are processed using Progenesis QI and the regulated proteins are subjected to METACORE, STRING, and WebGestalt for functional enrichment analysis. A total of 1614 proteins with ≥2 peptides are quantified with an estimated protein false discovery rate of 0.06%. Unsupervised clustering analysis shows that the gingival tissue protein abundance is mainly dependent on the periodontitis progression stage. Gene ontology enrichment analysis reveals an overrepresentation in innate immune regulation (e.g., neutrophil‐mediated immunity and antimicrobial peptides), signal transduction (e.g., integrin signaling), and homeostasis processes (e.g., platelet activation and aggregation). In conclusion, a PCT‐assisted label‐free quantitative proteomics workflow that allowed cataloging the deepest gingival tissue proteome on a rapid timescale and provided novel mechanistic insights into host perturbation during periodontitis progression is applied.     

Anoxia can increase the rate of decay for cnidarian tissue: Using Actinia equina to understand the early fossil record

An experimental decay methodology is developed for a cnidarian model organism to serve as a comparison to the many previous such studies on bilaterians. This allows an examination of inherent bias against the fossilisation of cnidarian tissue and their diagnostic characters, under what conditions these occur, and in what way. The decay sequence of Actinia equina was examined under a series of controlled conditions. These experiments show that cnidarian decay begins with an initial rupturing of the epidermis, followed by rapid loss of recognisable internal morphological characters. This suggests that bacteria work quicker on the epidermis than autolysis does on the internal anatomy. The data also show that diploblastic tissue is not universally decayed more slowly under anoxic or reducing conditions than under oxic conditions. Indeed, some cnidarian characters decay more rapidly under anoxic conditions than they do under oxic conditions. This suggests the decay pathways acting may be different to those affecting soft bilaterian tissue such as soft epidermis and internal organs. What is most important in the decay of soft polyp anatomy is the microbial community, which can be dominated by oxic or anoxic bacteria. Different Lagerstätte, even of the same type, will inevitably have subtle difference in their bacterial communities, which among other factors, could be a control on soft polyp preservation leading to either an absence of compelling soft anthozoans (Burgess Shale) or an astonishing abundance (Qingjiang biota).     

Experimental models of maternal–fetal interface and their potential use for nanotechnology applications

During pregnancy, the placenta regulates the transfer of oxygen, nutrients, and residual products between the maternal and fetal bloodstreams and is a key determinant of fetal exposure to xenobiotics from the mother. To study the disposition of substances through the placenta, various experimental models are used, especially the perfused placenta, placental villi explants, and cell lineage models. In this context, nanotechnology, an area of study that is on the rise, enables the creation of particles on nanometric scales capable of releasing drugs aimed at specific tissues. An important reason for furthering the studies on transplacental transfer is to explore the potential of nanoparticles (NPs), in new delivery strategies for drugs that are specifically aimed at the mother, the placenta, or the fetus and that involve less toxicity. Due to the fact that the placental barrier is essential for the interaction between the maternal and fetal organisms as well as the possibility of NPs being used in the treatment of various pathologies, the aim of this review is to present the main experimental models used in studying the maternal–fetal interaction and the action of NPs in the placental environment.     

The microevolutionary response to male‐limited X‐chromosome evolution in Drosophila melanogaster reflects macroevolutionary patterns

Due to its hemizygous inheritance and role in sex determination, the X‐chromosome is expected to play an important role in the evolution of sexual dimorphism and to be enriched for sexually antagonistic genetic variation. By forcing the X‐chromosome to only be expressed in males over >40 generations, we changed the selection pressures on the X to become similar to those experienced by the Y. This releases the X from any constraints arising from selection in females and should lead to specialization for male fitness, which could occur either via direct effects of X‐linked loci or trans‐regulation of autosomal loci by the X. We found evidence of masculinization via up‐regulation of male‐benefit sexually antagonistic genes and down‐regulation of X‐linked female‐benefit genes. Potential artefacts of the experimental evolution protocol are discussed and cannot be wholly discounted, leading to several caveats. Interestingly, we could detect evidence of microevolutionary changes consistent with previously documented macroevolutionary patterns, such as changes in expression consistent with previously established patterns of sexual dimorphism, an increase in the expression of metabolic genes related to mito‐nuclear conflict and evidence that dosage compensation effects can be rapidly altered. These results confirm the importance of the X in the evolution of sexual dimorphism and as a source for sexually antagonistic genetic variation and demonstrate that experimental evolution can be a fruitful method for testing theories of sex chromosome evolution.     

Spatial connectedness imposes local‐ and metapopulation‐level selection on life history through feedbacks on demography

Dispersal evolution impacts the fluxes of individuals and hence, connectivity in metapopulations. Connectivity is therefore decoupled from the structural connectedness of the patches within the spatial network. Because of demographic feedbacks, local selection also drives the evolution of other life history traits. We investigated how different levels of connectedness affect trait evolution in experimental metapopulations of the two‐spotted spider mite. We separated local‐ and metapopulation‐level selection and linked trait divergence to population dynamics. With lower connectedness, an increased starvation resistance and delayed dispersal evolved. Reproductive performance evolved locally by transgenerational plasticity or epigenetic processes. Costs of dispersal, but also changes in local densities and temporal fluctuations herein are found to be putative drivers. In addition to dispersal, demographic traits are able to evolve in response to metapopulation connectedness at both the local and metapopulation level by genetic and/or non‐genetic inheritance. These trait changes impact the persistence of spatially structured populations.     

以环境育人为理念的本科教学实验室管理模式探讨

本科教学实验室是开展本科生实验教学的主要场所,如何通过实验室的管理有效提升学生实验素养值得探索。文中主要介绍了将“6S (整理 (Seiri)、整顿 (Seiton)、清扫 (Seiso)、清洁 (Seiketsu)、素养 (Shitsuke)、安全 (Safety) 六项要素) 管理”理念引入本科教学实验室管理的应用实践与效果,特别是实验室环境管理、实验室安全管理、物品定位管理、试剂和耗材管理、学生培训和考核等方面的具体做法。通过实施“6S管理”,实验室环境得以改善,学生的实验素养得到提高,本科实验教学的质量得到改善和提高,达到环境育人的目的。这为高校教学实验室的建设和有效管理提供可推广、可操作的经验。     

不同品种玉米对草地贪夜蛾生长发育及繁殖的影响

为评价新入侵害虫草地贪夜蛾对我国不同品种玉米的为害潜能,本文采用组建实验种群生命表的方法研究了草地贪夜蛾对3种甜质型玉米(穗甜1号、正甜68和华金甜1号)以及3种糯质型玉米(京科糯2000、广黑甜糯和广糯1号)的生存适合度,分析了草地贪夜蛾生物学指标与玉米叶片主要营养物质及粗纤维含量间的相关性。结果表明: 取食甜质型玉米的草地贪夜蛾的幼虫存活率、蛹重、雌成虫产卵量均显著高于取食糯质型玉米;取食甜质型玉米草地贪夜蛾的内禀增长率介于0.1566～0.1843,净生殖力介于187.97～353.35,也均高于取食糯质型玉米(内禀增长率介于0.0998～0.1465,净生殖力介于25.89～95.34),表明取食甜质型玉米的草地贪夜蛾具有更高的种群增长能力。甜质型玉米叶片中主要营养物质维生素C、淀粉、可溶性糖、蛋白质、脂肪、总氨基酸及粗纤维的含量普遍高于糯质型玉米,草地贪夜蛾的净生殖力与维生素C、淀粉、可溶性糖、蛋白质及粗纤维的含量均存在显著正相关关系。草地贪夜蛾在甜质型玉米上具有更高的生存适合度。     

Prior evolution in stochastic versus constant temperatures affects RNA virus evolvability at a thermal extreme

It is unclear how historical adaptation versus maladaptation in a prior environment affects population evolvability in a novel habitat. Prior work showed that vesicular stomatitis virus (VSV) populations evolved at constant 37°C improved in cellular infection at both 29°C and 37°C; in contrast, those evolved under random changing temperatures between 29°C and 37°C failed to improve. Here, we tested whether prior evolution affected the rate of adaptation at the thermal‐niche edge: 40°C. After 40 virus generations in the new environment, we observed that populations historically evolved at random temperatures showed greater adaptability. Deep sequencing revealed that most of the newly evolved mutations were de novo. Also, two novel evolved mutations in the VSV glycoprotein and replicase genes tended to co‐occur in the populations previously evolved at constant 37°C, whereas this parallelism was not seen in populations with prior random temperature evolution. These results suggest that prior adaptation under constant versus random temperatures constrained the mutation landscape that could improve fitness in the novel 40°C environment, perhaps owing to differing epistatic effects of new mutations entering genetic architectures that earlier diverged. We concluded that RNA viruses maladapted to their previous environment could “leapfrog” over counterparts of higher fitness, to achieve faster adaptability in a novel environment.     

Evolution and maintenance of microbe‐mediated protection under occasional pathogen infection

Every host is colonized by a variety of microbes, some of which can protect their hosts from pathogen infection. However, pathogen presence naturally varies over time in nature, such as in the case of seasonal epidemics. We experimentally coevolved populations of Caenorhabditis elegans worm hosts with bacteria possessing protective traits (Enterococcus faecalis), in treatments varying the infection frequency with pathogenic Staphylococcus aureus every host generation, alternating host generations, every fifth host generation, or never. We additionally investigated the effect of initial pathogen presence at the formation of the defensive symbiosis. Our results show that enhanced microbe‐mediated protection evolved during host‐protective microbe coevolution when faced with rare infections by a pathogen. Initial pathogen presence had no effect on the evolutionary outcome of microbe‐mediated protection. We also found that protection was only effective at preventing mortality during the time of pathogen infection. Overall, our results suggest that resident microbes can be a form of transgenerational immunity against rare pathogen infection.