Use of the flexible film isolator as a single circuit hypoxic chamber for small animals

PVC isolators are now widely used for housing animals and provide a readily available pretested air-tight chamber (Pendry, 1984; Trexler, 1984). We have adapted a flexible film isolator for use as a hypoxic chamber for small animals. The environment within the chamber can be easily and continuously monitored with indwelling probes, obviating the need for a separate circuit for gas analysis. This design has been used for long-term studies of chronic hypoxia.     

Effects of pesticides on spin down and webbing production by the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae)

Spin down in response to pesticides is a known phenomenon. This study aimed at investigating the effects of varying acaricide concentrations and acaricidal combinations on the rate of spin down and thread length, the fate of mites after spinning down and, thus, the implications of spin down in the context of mite control and the relationship between webbing and the efficacy of ovicides. In response to cyhexatin at a concentration of up to 100 mg dm-3 the rate of spin down did not change, but in response to higher concentrations it occurred earlier and ceased before the end of the experiment. The mean thread length did not vary with the concentration of acaricide and, whilst the proportion of mites spinning down increased with a cyhexatin concentration of up to 10 mg dm-3, it did not increase further with higher concentrations thereafter. It is therefore postulated that at any given time only a certain proportion of mites have a propensity to spin down. In combination with cyhexatin, only amitraz (at low concentrations) increased the proportion of mites spinning down, but amitraz (at higher concentrations), flubenzimine and permethrin all inhibited spin down, which may be a useful attribute in pesticide formulations. With regard to the fate of mites after spinning down, they were able to climb back up threads, but the proportion doing so was too small to be of practical importance. Mites which spin down and remain at the end of a thread die, but those which reached another leaf were able to spin down again and were more likely to do so if the leaf bore a cyhexatin residue. Mites reaching another leaf are also able to lay eggs, the number being greater if the leaf which they reach is untreated. Spin down is therefore likely to be of significant advantage to Tetranychus urticae in avoidance of the effects of acaricides. Finally, some eggs were observed to be laid on webbing and not to come into direct contact with clofentezine residues. The use of a fine water spray to simulate dew increased the ovitoxicity of clofentezine, probably by resuspension of the pesticide, but not of amitraz, which inhibits webbing production. The avoidance of acaricides should be added to the several known advantages of webbing production for spider mites. It is concluded that, all other factors being equal, pesticide formulations which inhibit spin down and webbing production are likely to be more effective in control programmes than those which stimulate such behaviour.     

CHOmpact: A reduced metabolic model of Chinese hamster ovary cells with enhanced interpretability

Metabolic modeling has emerged as a key tool for the characterization of biopharmaceutical cell culture processes. Metabolic models have also been instrumental in identifying genetic engineering targets and developing feeding strategies that optimize the growth and productivity of Chinese hamster ovary (CHO) cells. Despite their success, metabolic models of CHO cells still present considerable challenges. Genome-scale metabolic models (GeMs) of CHO cells are very large (>6000 reactions) and are difficult to constrain to yield physiologically consistent flux distributions. The large scale of GeMs also makes the interpretation of their outputs difficult. To address these challenges, we have developed CHOmpact, a reduced metabolic network that encompasses 101 metabolites linked through 144 reactions. Our compact reaction network allows us to deploy robust, nonlinear optimization and ensure that the computed flux distributions are physiologically consistent. Furthermore, our CHOmpact model delivers enhanced interpretability of simulation results and has allowed us to identify the mechanisms governing shifts in the anaplerotic consumption of asparagine and glutamate as well as an important mechanism of ammonia detoxification within mitochondria. CHOmpact, thus, addresses key challenges of large-scale metabolic models and will serve as a platform to develop dynamic metabolic models for the control and optimization of biopharmaceutical cell culture processes.     

GeneScape: a relational database of Escherichia coli genomic map data for Macintosh computers

We present a relational database program developed in FoxBase+/Mac for the viewing and manipulation of ordered restrictionmaps and associated features of the Escherichia coli genomeincluding scquenced genes and the Kohara miniset of bacteriophagelambda clones. Use of this program allows easy access to thewealth of information being collected in a datase ofDNA sequences,maps and genetic data known as EcoSeq, EcoMap and EcoGene respectively.     

Factors distinguishing couples at risk for nondisjunction

Micronucleus frequencies were determined on 24 young parents of trisomic infants, 21 individuals with recurrent unexplained abortions, and 42 control individuals with proven reproductive success. In addition to measurements of spontaneous micronucleus frequencies, mitomycin C induced frequencies were determined at two doses (2.5 and 5.0 ng/mL). Using the 95% confidence limits established from control data as an arbitrary upper limit, 16 of 24 parents of trisomics and 5 of 21 recurrent aborters were detected by the micronucleus assay to above this cutoff. The effects of sex, age, pregnancy status, and a variety of environmental exposures were studied by comparing the micronucleus frequencies of the exposed and unexposed populations. The data suggested that vitamins were associated with a lower micronucleus frequency and tea drinking with an increased micronucleus frequency in parents of trisomics, an effect not seen in controls. These results suggest that a biologic basis for nondisjunction may be associated with elevation in spontaneous and induced micronuclei. In addition, tea and vitamins may modulate micronucleus frequencies in parents of trisomics who appear to be more sensitive to these influences than the controls.     

Remarkable recent changes in the genetic diversity of the avirulence gene AvrStb6 in global populations of the wheat pathogen Zymoseptoria tritici

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is one of the most economically important diseases of wheat. Recently, both factors of a gene-for-gene interaction between Z. tritici and wheat, the wheat receptor-like kinase Stb6 and the Z. tritici secreted effector protein AvrStb6, have been identified. Previous analyses revealed a high diversity of AvrStb6 haplotypes present in earlier Z. tritici isolate collections, with up to c.18% of analysed isolates possessing the avirulence isoform of AvrStb6 identical to that originally identified in the reference isolate IPO323. With Stb6 present in many commercial wheat cultivars globally, we aimed to assess potential changes in AvrStb6 genetic diversity and the incidence of haplotypes allowing evasion of Stb6-mediated resistance in more recent Z. tritici populations. Here we show, using targeted resequencing of AvrStb6, that this gene is universally present in field isolates sampled from major wheat-growing regions of the world in 2013–2017. However, in contrast to the data from previous AvrStb6 population studies, we report a complete absence of the originally described avirulence isoform of AvrStb6 amongst modern Z. tritici isolates. Moreover, a remarkably small number of haplotypes, each encoding AvrStb6 protein isoforms conditioning virulence on Stb6-containing wheat, were found to predominate among modern Z. tritici isolates. A single virulence isoform of AvrStb6 was found to be particularly abundant throughout the global population. These findings indicate that, despite the ability of Z. tritici to sexually reproduce on resistant hosts, AvrStb6 avirulence haplotypes tend to be eliminated in subsequent populations.     

Erythrapheresis in patients with polycythaemia secondary to hypoxic lung disease.

Erythrapheresis was performed in 10 patients with polycythaemia secondary to hypoxic lung disease (mean PaO2, 6.8 kPa (51 mm Hg)). The mean packed cell volume decreased from 0.64 to 0.48 in men and from 0.56 to 0.42 in women, with significant decreases in blood viscosity at both high and low shear rates (p less than 0.001). Patients showed significant improvement in six-minute walking distances (p less than 0.001) and in tests of mental alertness (p less than 0.01) compared to control subjects. Visual analogue scales confirmed symptomatic improvement after erythrapheresis. Erythrapheresis significantly improved symptoms, mental function, and work performance in patients with polycythaemia secondary to hypoxic lung disease. The procedure was well tolerated by all patients and no complications occurred.     

Epigenetic regulation of glycosylation and the impact on chemo-resistance in breast and ovarian cancer

Glycosylation is one of the most fundamental posttranslational modifications in cellular biology and has been shown to be epigenetically regulated. Understanding this process is important as epigenetic therapies such as those using DNA methyltransferase inhibitors are undergoing clinical trials for the treatment of ovarian and breast cancer. Previous work has demonstrated that altered glycosylation patterns are associated with aggressive disease in women presenting with breast and ovarian cancer. Moreover, the tumor microenvironment of hypoxia results in globally altered DNA methylation and is associated with aggressive cancer phenotypes and chemo-resistance, a feature integral to many cancers. There is sparse knowledge on the impact of these therapies on glycosylation. Moreover, little is known about the efficacy of DNA methyltransferase inhibitors in hypoxic tumors. In this review, we interrogate the impact that hypoxia and epigenetic regulation has on cancer cell glycosylation in relation to resultant tumor cell aggressiveness and chemo-resistance.