Composition of the nucleotides pool in a morphogenetic compartment in eggs of Nassarius reticulatus (mollusca) analysed by capillary isotachophoresis

The spectrum of low molecular weight compounds, in particular of ribonucleotides, within first cleavage stage embryos of the polar lobe-forming mollusc Nassarius reticulatus and the distribution of the compounds within the embryo at the trefoil stage of first cleavage are analysed by means of capillary isotachophoresis after 0.5 M PCA extraction. The compounds which are found in the whole trefoil embryo (T), the lobeless part (LL), and the polar lobe (PL) respectively, and the mean quantities (nmol. microliter-1; n = 6) are: UTP (11.5, 4.8, 5.6), ITP (8.5, 3.6, 5.0), GTP (10.3, 3.0, 9.0), ATP (29.8, 13.4, 18.8), UDP (11.8, 3.4, 8.7), CTP (8.0, 3.1, 4.5), GDP (5.3, 2.6, 3.4), ADP (16.5, 6.1, 11.6), CDP (4.0, 1.4, 2.6), GMP (4.7, 2.7, 4.3), glucose-6-phosphate (G6P) (53.5, 38.8, 13.0). These compounds appear to be localized in the non-yolk cytoplasmic pool. As the volume ratio of PL/LL for total volume and for non-yolk cytoplasmic volume is about 0.74 and 0.60 respectively, the concentration of all nucleotides in PL as compared to LL is significantly higher (HO, p less than 0.001), both relative to the total volume and to the non-yolk cytoplasmic volume. The G6P concentration is considerably higher in the lobeless part. The morphogenetic role of the vegetal pole compartment of the egg apparently is correlated with a relatively high level of its nucleotide contents.     

Until death do us part? In-depth insights into Dutch consumers’ considerations about product lifetimes and lifetime extension

Long-lasting electronic products contribute to a sustainable society; however, both expected and actual lifetimes are in decline. This research provides in-depth insights into consumers’ considerations about product lifetimes, barriers to extending lifetimes, and responses to a product lifetime label. Results of interviews (n = 22) with Dutch consumers suggest a positive view on long-lasting products. Nevertheless, their products’ value depreciated during their lifetimes. Consumers consider themselves unable to estimate how long products should last, which can be detrimental as low expectations tend to negatively influence actual lifetimes. Also, use intensity and consumers’ care(less) behavior influence the lifetime. To extend product lifetimes, consumers often disregard the option of repairing malfunctioning products. They have limited knowledge and ability, and believe repair provides poor value for money. Lifetime extension can also be hindered by market-related factors, such as convenient replacement services, new technological developments, and (attractive) deals. We suggest a product lifetime label should contain relevant and reliable information; furthermore, we recommend including (extended) warranty information. When information about repairability is included, potential negative responses should be considered. Finally, raising awareness about the environmental impact of short-lived products via a label may have a positive effect but requires more research attention.     

Vasopressin levels in peripheral blood and in cerebrospinal fluid during passive and active avoidance behavior in rats.

Vasopressin (AVP) levels were measured in plasma and cerebrospinal fluid (CSF) of rats during acquisition and retention of a passive avoidance response. Only 5 min after the onset of the retention session a significantly higher level of AVP was found in plasma of animals which displayed a long latency, as compared with the levels of animals which showed a weak passive avoidance response (short latencies), or no passive avoidance behavior at all (controls). Moreover no changes in plasma AVP levels were found in plasma of rats submitted to acquisition or extinction of an active avoidance response. It is suggested that, although an elevated plasma AVP level is associated with strong retention of a passive avoidance response the peripheral circulation as well as the CSF are of minor importance for the transport of this neuropeptide to its site of behavioral action.     

Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine

Crosslink repair depends on the Fanconi anemia pathway and translesion synthesis polymerases that replicate over unhooked crosslinks. Translesion synthesis is regulated via ubiquitination of PCNA, and independently via translesion synthesis polymerase REV1. The division of labor between PCNA-ubiquitination and REV1 in interstrand crosslink repair is unclear. Inhibition of either of these pathways has been proposed as a strategy to increase cytotoxicity of platinating agents in cancer treatment. Here, we defined the importance of PCNA-ubiquitination and REV1 for DNA in mammalian ICL repair. In mice, loss of PCNA-ubiquitination, but not REV1, resulted in germ cell defects and hypersensitivity to cisplatin. Loss of PCNA-ubiquitination, but not REV1 sensitized mammalian cancer cell lines to cisplatin. We identify polymerase Kappa as essential in tolerating DNA damage-induced lesions, in particular cisplatin lesions. Polk-deficient tumors were controlled by cisplatin treatment and it significantly delayed tumor outgrowth and increased overall survival of tumor bearing mice. Our results indicate that PCNA-ubiquitination and REV1 play distinct roles in DNA damage tolerance. Moreover, our results highlight POLK as a critical TLS polymerase in tolerating multiple genotoxic lesions, including cisplatin lesions. The relative frequent loss of Polk in cancers indicates an exploitable vulnerability for precision cancer medicine.     

Diversity of an ectomycorrhizal fungal community studied by a root tip and total soil DNA approach

Molecular methods based on soil DNA extracts are increasingly being used to study the fungal diversity of ectomycorrhizal (EM) fungal communities in soil. Contrary to EM root tip identification, the use of molecular methods enables identification of extramatrical mycelia in soil. To compare fungal diversity as determined by root tip identification and mycelial identification, six soil samples were analysed. Root tips were extracted from the six samples and after amplification, the basidiomycete diversity on the root tips was analysed by denaturing gradient gel electrophoresis (DGGE). The soil from the six samples was sieved, total soil DNA was extracted and after amplification, the basidiomycete diversity in the soil fractions was analysed by DGGE. Fourteen different bands were excised from the DGGE gel and sequenced; fungal taxon names could be assigned to eight bands. Out of a total of 14 fungal taxa detected in soil, 11 fungal taxa were found on root tips, of which seven were EM fungal taxa. To examine whether the sieving treatment would affect EM species diversity, two different sieve mesh sizes were used and in addition, the organic soil fraction was analysed separately. DGGE analysis showed no differences in banding pattern for the different soil fractions. The organic fraction gave the highest DGGE band intensities. This work demonstrates that there is a high correspondence between basidiomycete diversity detected by molecular analysis of root tips and soil samples, irrespective of the soil fraction being analysed.     

Plant community composition steers grassland vegetation via soil legacy effects

Soil legacy effects are commonly highlighted as drivers of plant community dynamics and species co‐existence. However, experimental evidence for soil legacy effects of conditioning plant communities on responding plant communities under natural conditions is lacking. We conditioned 192 grassland plots using six different plant communities with different ratios of grasses and forbs and for different durations. Soil microbial legacies were evident for soil fungi, but not for soil bacteria, while soil abiotic parameters did not significantly change in response to conditioning. The soil legacies affected the composition of the succeeding vegetation. Plant communities with different ratios of grasses and forbs left soil legacies that negatively affected succeeding plants of the same functional type. We conclude that fungal‐mediated soil legacy effects play a significant role in vegetation assembly of natural plant communities.     

An Evaluation of Plotless Sampling Using Vegetation Simulations and Field Data from a Mangrove Forest

In vegetation science and forest management, tree density is often used as a variable. To determine the value of this variable, reliable field methods are necessary. When vegetation is sparse or not easily accessible, the use of sample plots is not feasible in the field. Therefore, plotless methods, like the Point Centred Quarter Method, are often used as an alternative. In this study we investigate the accuracy of different plotless sampling methods. To this end, tree densities of a mangrove forest were determined and compared with estimates provided by several plotless methods. None of these methods proved accurate across all field sites with mean underestimations up to 97% and mean overestimations up to 53% in the field. Applying the methods to different vegetation patterns shows that when random spatial distributions were used the true density was included within the 95% confidence limits of all the plotless methods tested. It was also found that, besides aggregation and regularity, density trends often found in mangroves contribute to the unreliability. This outcome raises questions about the use of plotless sampling in forest monitoring and management, as well as for estimates of density-based carbon sequestration. We give recommendations to minimize errors in vegetation surveys and recommendations for further in-depth research.     

Ex vivo Live Imaging of Lung Metastasis and Their Microenvironment

Metastasis is a major cause for cancer-related morbidity and mortality. Metastasis is a multistep process and due to its complexity, the exact cellular and molecular processes that govern metastatic dissemination and growth are still elusive. Live imaging allows visualization of the dynamic and spatial interactions of cells and their microenvironment. Solid tumors commonly metastasize to the lungs. However, the anatomical location of the lungs poses a challenge to intravital imaging. This protocol provides a relatively simple and quick method for ex vivo live imaging of the dynamic interactions between tumor cells and their surrounding stroma within lung metastasis. Using this method, the motility of cancer cells as well as interactions between cancer cells and stromal cells in their microenvironment can be visualized in real time for several hours. By using transgenic fluorescent reporter mice, a fluorescent cell line, injectable fluorescently labeled molecules and/or antibodies, multiple components of the lung microenvironment can be visualized, such as blood vessels and immune cells. To image the different cell types, a spinning disk confocal microscope that allows long-term continuous imaging with rapid, four-color image acquisition has been used. Time-lapse movies compiled from images collected over multiple positions and focal planes show interactions between live metastatic and immune cells for at least 4 hr. This technique can be further used to test chemotherapy or targeted therapy. Moreover, this method could be adapted for the study of other lung-related pathologies that may affect the lung microenvironment.