Deciphering the Link between Doubly Uniparental Inheritance of mtDNA and Sex Determination in Bivalves: Clues from Comparative Transcriptomics

Bivalves exhibit an astonishing diversity of sexual systems and sex-determining mechanisms. They can be gonochoric, hermaphroditic or androgenetic, with both genetic and environmental factors known to determine or influence sex. One unique sex-determining system involving the mitochondrial genome has also been hypothesized to exist in bivalves with doubly uniparental inheritance (DUI) of mtDNA. However, the link between DUI and sex determination remains obscure. In this study, we performed a comparative gonad transcriptomics analysis for two DUI-possessing freshwater mussel species to better understand the mechanisms underlying sex determination and DUI in these bivalves. We used a BLAST reciprocal analysis to identify orthologs between Venustaconcha ellipsiformis and Utterbackia peninsularis and compared our results with previously published sex-specific bivalve transcriptomes to identify conserved sex-determining genes. We also compared our data with other DUI species to identify candidate genes possibly involved in the regulation of DUI. A total of 3c12,000 orthologous relationships were found, with 2,583 genes differentially expressed in both species. Among these genes, key sex-determining factors previously reported in vertebrates and in bivalves (e.g., Sry, Dmrt1, Foxl2) were identified, suggesting that some steps of the sex-determination pathway may be deeply conserved in metazoans. Our results also support the hypothesis that a modified ubiquitination mechanism could be responsible for the retention of the paternal mtDNA in male bivalves, and revealed that DNA methylation could also be involved in the regulation of DUI. Globally, our results suggest that sets of genes associated with sex determination and DUI are similar in distantly-related DUI species

The absence of clinical disease in cattle in communal grazing areas where farmers are changing from an intensive dipping programme to one of endemic stability to tick-borne diseases

A two-year field study was conducted in four communal grazing areas in South Africa. Sera were collected from young cattle (6-18 months old) in these areas during the winters of 1991 to 1993. The sera were tested for antibodies to Babesia bovis, Babesia bigemina, Anaplasma marginale and Cowdria ruminantium. In two of the four areas, treatment with acaricide was erratic and dependent on the discretion of individual owners. In these areas the drought of 1992 had a major impact on tick burdens and there were changes in the seroprevalence to tick-borne diseases. In the other two areas there was a reduction in the intensity of acaricide application and this was associated with an increase in seropositivity to the tick-borne diseases. Increases in the prevalence of seropositivity and the presence of endemic instability, as calculated from inoculation rates, were not accompanied by outbreaks of clinical disease. Possible reasons for this are discussed.The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat X Pro was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.The Medical University of Southern Africa (MEDUNSA). Foundation for Research and Development (University Development Programme).mn201

Efficient Parallel Statistical Model Checking of Biochemical Networks

We consider the problem of verifying stochastic models of biochemical networks against behavioral properties expressed in temporal logic terms. Exact probabilistic verification approaches such as, for example, CSL/PCTL model checking, are undermined by a huge computational demand which rule them out for most real case studies. Less demanding approaches, such as statistical model checking, estimate the likelihood that a property is satisfied by sampling executions out of the stochastic model. We propose a methodology for efficiently estimating the likelihood that a LTL property P holds of a stochastic model of a biochemical network. As with other statistical verification techniques, the methodology we propose uses a stochastic simulation algorithm for generating execution samples, however there are three key aspects that improve the efficiency: first, the sample generation is driven by on-the-fly verification of P which results in optimal overall simulation time. Second, the confidence interval estimation for the probability of P to hold is based on an efficient variant of the Wilson method which ensures a faster convergence. Third, the whole methodology is designed according to a parallel fashion and a prototype software tool has been implemented that performs the sampling/verification process in parallel over an HPC architecture

Cell-free protein synthesis of membrane (1,3)-beta-D-glucan (curdlan) synthase: Co-translational insertion in liposomes and reconstitution in nanodiscs

A membrane-embedded curdlan synthase (CrdS) from Agrobacterium is believed to catalyse a repetitive addition of glucosyl residues from UDP-glucose to produce the (1,3)-β-d-glucan (curdlan) polymer. We report wheat germ cell-free protein synthesis (WG-CFPS) of full-length CrdS containing a 6xHis affinity tag and either Factor Xa or Tobacco Etch Virus proteolytic sites, using a variety of hydrophobic membrane-mimicking environments. Full-length CrdS was synthesised with no variations in primary structure, following analysis of tryptic fragments by MALDI-TOF/TOF Mass Spectrometry. Preparative scale WG-CFPS in dialysis mode with Brij-58 yielded CrdS in mg/ml quantities. Analysis of structural and functional properties of CrdS during protein synthesis showed that CrdS was co-translationally inserted in DMPC liposomes during WG-CFPS, and these liposomes could be purified in a single step by density gradient floatation. Incorporated CrdS exhibited a random orientation topology. Following affinity purification of CrdS, the protein was reconstituted in nanodiscs with Escherichia coli lipids or POPC and a membrane scaffold protein MSP1E3D1. CrdS nanodiscs were characterised by small-angle X-ray scattering using synchrotron radiation and the data obtained were consistent with insertion of CrdS into bilayers. We found CrdS synthesised in the presence of the Ac-AAAAAAD surfactant peptide or co-translationally inserted in liposomes made from E. coli lipids to be catalytically competent. Conversely, CrdS synthesised with only Brij-58 was inactive. Our findings pave the way for future structural studies of this industrially important catalytic membrane protein.Agalya Periasamy, Nadim Shadiac, Amritha Amalraj, Soňa Garajová, Yagnesh Nagarajan, Shane Waters, Haydyn D.T. Mertens, Maria Hrmov

Targeting the Ataxia Telangiectasia Mutated-null Phenotype in Chronic Lymphocytic Leukemia with Pro-oxidants

Inactivation of the Ataxia Telangiectasia Mutated gene in chronic lymphocytic leukemia results in resistance to p53-dependent apoptosis and inferior responses to treatment with DNA damaging agents. Hence, p53-independent strategies are required to target Ataxia Telangiectasia Mutated-deficient chronic lymphocytic leukemia. As Ataxia Telangiectasia Mutated has been implicated in redox homeostasis, we investigated the effect of the Ataxia Telangiectasia Mutated-null chronic lymphocytic leukemia genotype on cellular responses to oxidative stress with a view to therapeutic targeting. We found that in comparison to Ataxia Telangiectasia Mutated-wild type chronic lymphocytic leukemia, pro-oxidant treatment of Ataxia Telangiectasia Mutated-null cells led to reduced binding of NF-E2 p45-related factor-2 to antioxidant response elements and thus decreased expression of target genes. Furthermore, Ataxia Telangiectasia Mutated-null chronic lymphocytic leukemia cells contained lower levels of antioxidants and elevated mitochondrial reactive oxygen species. Consequently, Ataxia Telangiectasia Mutated-null chronic lymphocytic leukemia, but not tumours with 11q deletion or TP53 mutations, exhibited differentially increased sensitivity to pro-oxidants both in vitro and in vivo. We found that cell death was mediated by a p53- and caspase-independent mechanism associated with apoptosis inducing factor activity. Together, these data suggest that defective redox-homeostasis represents an attractive therapeutic target for Ataxia Telangiectasia Mutated-null chronic lymphocytic leukemia

Risk Owners & Risk Managers: Dealing with the complexity of feeding children with neurodevelopmental disability

This paper illustrates negotiations around risk between lay people and clinicians in relation to gastrostomy interventions for disabled children. These negotiations centre on differing interpretations of what constitutes risk in relation to the safety of oral feeding and a child's need for a feeding tube between parents, carers and clinical specialties. Drawing on Heyman's distinction between risk managers and risk owners, we show that not only do clinicians act as risk managers and parents and carers as risk owners, but that these distinctions often become blurred either because of the shifting dynamics of relations of care or because of the specificity of clinical practice. Parents become risk managers in relation to carers' roles, while clinicians become risk owners in relation to particular procedures which define their practice. This has implications for lay and expert interactions as well as professional accountability for those caring for children with complex medical conditions. Although not an empirical article, we draw on empirical work in the UK. We analyse both parental and professional constructions of risk based on observations of co-ordinating a clinical trial designed to evaluate the effectiveness of gastrostomy surgery. We also examine the diverse value systems used by different groups of professionals and lay carers which inform judgements about risk and feeding. We conclude by arguing that issues of risk in contemporary health care are not just examples of ‘manufactured uncertainty’ or of ‘negotiated power’ but constitute a dialectical relationship which breaks down the essentialist dualism of lay and professional constructions of risk

Reply to editorial and commentaries on Steele, Al-Mufti, Augustyn, Chandrajith, Coghlan, Coulson et al. (2018) "Cause of Cambrian explosion - Terrestrial or cosmic?"

No abstract availabl

Reply to commentary by R Duggleby (2019)

Duggleby (2018) has made a numerical analysis of some aspects of the wide range of phenomena we reviewed in Steele et al. (2018) and asserted " .that panspermia as proposed by Steele et al. (2018) is extremely implausible.” It seems to us that Duggleby has based his viewpoint on a quite narrow and specific model of Panspermia which he supposes to be active in the cosmos. Here we address both his conclusions and his numerical analysis. Our response therefore will be at two levels, his specific analysis and his general conclusions. In the specific section below we show that while Duggleby's numerical analysis appears in part correct it is, in the final analysis, quite irrelevant to Cosmic Panspermia. In the general response which follows we address his unsupported conclusion throughout his critique, namely that … " none of the examples mentioned by Steele et al. (2018) is decisive enough to allow no other explanation.

Thermodynamic Measurements in a Strongly Interacting Fermi Gas

We conduct a series of measurements on the thermodynamic properties of an optically-trapped strongly interacting Fermi gas, including the energy $E$, entropy $S$, and sound velocity $c$. Our model-independent measurements of $E$ and $S$ enable a precision study of the finite temperature thermodynamics. The $E(S)$ data are directly compared to several recent predictions. The temperature in both the superfluid and normal fluid regime is obtained from the fundamental thermodynamic relation $T=\partial E/\partial S$ by parameterizing the $E(S)$ data. Our $E(S)$ data are also used to experimentally calibrate the endpoint temperatures obtained for adiabatic sweeps of the magnetic field between the ideal and strongly interacting regimes. This enables the first experimental calibration of the temperature scale used in experiments on fermionic pair condensation. Our calibration shows that the ideal gas temperature measured for the onset of pair condensation corresponds closely to the critical temperature estimated in the strongly interacting regime from the fits to our $E(S)$ data. The results are in very good agreement with recent predictions. Finally, using universal thermodynamic relations, we estimate the chemical potential and heat capacity of the trapped gas from the $E(S)$ data.Comment: 29 pages, 12 figures. To appear in JLTP online, and in the January, 2009 volum