Urinary bisphenol a concentration and angiography-defined coronary artery stenosis

Background

Bisphenol A is widely used in food and drinks packaging. There is evidence of associations between raised urinary bisphenol A (uBPA) and increased incidence of reported cardiovascular diagnoses.

Methodology/Principal Findings

To estimate associations between BPA exposure and angiographically graded coronary atherosclerosis. 591 patients participating in The Metabonomics and Genomics in Coronary Artery Disease (MaGiCAD) study in Cambridgeshire UK, comparing urinary BPA (uBPA) with grades of severity of coronary artery disease (CAD) on angiography. Linear models were adjusted for BMI, occupational social class and diabetes status. Severe (one to three vessel) CAD was present in 385 patients, 86 had intermediate disease (n = 86) and 120 had normal coronary arteries. The (unadjusted) median uBPA concentration was 1.28 ng/mL with normal coronary arteries, and 1.53 ng/mL with severe CAD. Compared to those with normal coronary arteries, uBPA concentration was significantly higher in those with severe CAD (OR per uBPA SD = 5.96 ng/ml OR = 1.43, CI 1.03 to 1.98, p = 0.033), and near significant for intermediate disease (OR = 1.69, CI 0.98 to 2.94, p = 0.061). There was no significant uBPA difference between patients with severe CAD (needing surgery) and the remaining groups combined.

Conclusions/Significance

BPA exposure was higher in those with severe coronary artery stenoses compared to those with no vessel disease. Larger studies are needed to estimate true dose response relationships. The mechanisms underlying the association remain to be established.     

Homologous recombination properties of OsRad51, a recombinase from rice

cDNA corresponding to OsRad51 protein was isolated from cDNA library of rice flowers (Oryza sativa, Indica cultivar group) and cloned in to pET28a expression vector. The protein was over expressed in E. coli BL21 (DE3) and purified. Purified OsRad51 could bind single and double stranded DNA, however it showed higher affinity for single stranded DNA. Transmission Electron Microscopy (TEM) studies of OsRad51-DNA complexes showed that this protein formed ring like structures and bound DNA forming filaments. OsRad51 protein promoted renaturation of complementary single strands in to duplex DNA molecules and also showed ATPase activity, which was stimulated by single strand DNA. Fluorescence resonance energy transfer (FRET) assays revealed that OsRad51 promoted homology dependent renaturation as well as strand exchange reactions. Renaturation activity was ATP dependent; however strand exchange activity was ATP independent. This is the first report on in vitro characterization of Rad51 protein from crop plants.     

Choline transporter‐like1 (CHER1) is crucial for plasmodesmata maturation in Arabidopsis thaliana

Plasmodesmata (PD) are microscopic pores connecting plant cells and enable cell‐to‐cell transport. Currently, little information is known about the molecular mechanisms regulating PD formation and development. To uncover components of PD development we made use of the 17 kDa movement protein (MP17) encoded by the Potato leafroll virus (PLRV). The protein is required for cell‐to‐cell movement of the virus and localises to complex PD. Forward genetic screening for Arabidopsis mutants with altered PD binding of MP17 revealed several mutant lines, while molecular genetics, biochemical and microscopic studies allowed further characterisation. Map‐based cloning of one mutant revealed a point mutation in the choline transporter‐like 1 (CHER1) protein, changing glycine²⁴⁷ into glutamate. Mutation in CHER1 resulted in a starch excess phenotype and stunted growth. Ultrastructure analysis of shoot apical meristems, developing and fully developed leaves showed reduced PD numbers and the absence of complex PD in fully developed leaves. This indicates that cher1 mutants are impaired in PD formation and development. Global lipid profiling revealed only slight modifications in the overall lipid composition, however, altered composition of PD‐associated lipids cannot be ruled out. Thus, cher1 is devoid of complex PD in developed leaves and provides insights into the formation of complex PD at the molecular level.     

The Triangulation WIthin a STudy (TWIST) framework for causal inference within pharmacogenetic research

In this paper we review the methodological underpinnings of the general pharmacogenetic approach for uncovering genetically-driven treatment effect heterogeneity. This typically utilises only individuals who are treated and relies on fairly strong baseline assumptions to estimate what we term the ‘genetically moderated treatment effect’ (GMTE). When these assumptions are seriously violated, we show that a robust but less efficient estimate of the GMTE that incorporates information on the population of untreated individuals can instead be used. In cases of partial violation, we clarify when Mendelian randomization and a modified confounder adjustment method can also yield consistent estimates for the GMTE. A decision framework is then described to decide when a particular estimation strategy is most appropriate and how specific estimators can be combined to further improve efficiency. Triangulation of evidence from different data sources, each with their inherent biases and limitations, is becoming a well established principle for strengthening causal analysis. We call our framework ‘Triangulation WIthin a STudy’ (TWIST)’ in order to emphasise that an analysis in this spirit is also possible within a single data set, using causal estimates that are approximately uncorrelated, but reliant on different sets of assumptions. We illustrate these approaches by re-analysing primary-care-linked UK Biobank data relating to CYP2C19 genetic variants, Clopidogrel use and stroke risk, and data relating to APOE genetic variants, statin use and Coronary Artery Disease.     

Development of the sensory system in larvae and pupae of Chaoborus crystallinus (DeGeer, 1776; Diptera, Chaoboridae): sensory cells, nerves and ganglia of the tail region

Using various microscopical techniques, we have studied changes in the sensory equipment and architecture of the peripheral nervous system (PNS) around the first metamorphic molt from larva to pupa in the phantom midge Chaoborus. The transparent larvae and pupae of this dipteran with ancestral features allow us to investigate sensilla and their central projections from whole-mount preparations of complete groups of segments. Each sensillum on the posterior larval and pupal segments was identified using its external shape and position, and the morphology of the abdominal ganglia and segmental nerves was investigated. In addition, retrograde fills with the carbocyanine dye DiI were used to trace the axonal paths of most of the extero- and proprioreceptors. These findings were combined to produce maps of the sensory elements of larval and pupal abdomens that were analyzed at three levels: seriality (homonomy), ontogenetic changes of individual sensilla, and homology of the PNS between different species. Comparison of different segments shows for both stages that primarily there is a homonomous basic design of the PNS, but segment-specific modifications are evident in segments 8-10. Comparison of corresponding larval and pupal segments shows that many sensilla retain their internal structure and axonal projections. However, their external cuticular parts are changed in relation to the different life habits of larvae and pupae. Furthermore, some sensilla are completely reduced during the pupal molt, especially those of the tenth segment which appears as a distinct larval structure (caenogenesis). Comparison between species indicates that despite the varying types of sensilla their basic segmental arrangement and their axonal trajectories are conserved.     

Simple sequence repeat (SSR) markers survey of the cassava (Manihot esculenta Crantz) genome: towards an SSR-based molecular genetic map of cassava

The development of PCR-based, easily automated molecular genetic markers, such as SSR markers, are required for realistic cost-effective marker-assisted selection schemes. This paper describes the development and characterization of 172 new SSR markers for the cassava genome. The placement of 36 of these markers on the existing RFLP framework map of cassava is also reported. Two similar enrichment methods were employed. The first method yielded 35 SSR loci, for which primers could be designed, out of 148 putative DNA clones. A total of 137 primer pairs could be designed from 544 putative clones sequenced for the second enrichment. Most of the SSRs (95%) were di-nucleotide repeats, and 21% were compound repeats. A major drawback of these methods of SSR discovery is the redundancy – 20% duplication; in addition, primers could not be designed for many SSR loci that were too close to the cloning site – 45% of the total. All 172 SSRs amplified the corresponding loci in the parents of the mapping progeny, with 66% of them revealing a unique allele in at least one of the parents, and 26% having unique alleles in both of the parents. Of the 36 SSRs that have been mapped, at least 1 was placed on 16 out of the 18 linkage groups of the framework map, indicating a broad coverage of the cassava genome. This preliminary mapping of the 36 markers has led to the joining of a few small groups and the creation of one new group. The abundance of allelic bridges as shown by these markers will lead to the development of a consensus map of the male- and female-derived linkage groups. In addition, the relatively higher number of these allelic bridges, 30% as against 10% for RFLPs in cassava, underscores SSR as the marker of choice for cassava. The 100% primer amplification obtained for this set of primers also confirms the appropriateness of SSR markers for use in cassava genome analysis and the transferability of the technology as a low-cost approach to increasing the efficiency of cassava breeding. Current efforts are geared towards the generation of more SSR markers to attain a goal of 200 SSR markers, or 1 SSR marker every 10 cM. Received: 15 November 1999 / Accepted: 14 April 2000     

The Trophic Index of Macrophytes (TIM) – a New Tool for Indicating the Trophic State of Running Waters

In running waters, apart from structural degradation, nutrient input becomes increasingly important. To investigate the indicator values of as many species of submerged macrophytes as possible numerous samples of the sediment within macrophyte stands and the overlying water were taken in running waters throughout Bavaria, Germany. To develop the Trophic Index of Macrophytes (TIM), the concentrations of soluble reactive phosphorus of both the water body and the sediment pore water were used. Based on a weighted sum of the SRP‐concentrations of the water body and the sediment pore water, indicator values were determined for a total of 49 species of submerged macrophytes. A detailed method is described on how and depending on which preconditions the trophic state of running waters can be determined by the TIM. An example of the TIM in the stream Rotbach is given. It shows that the TIM is a useful means to detect differences in the phosphorus loading of running waters.     

The effect of plantar flexor muscle fatigue on postural control

Objective

Previous studies have demonstrated that ankle muscle fatigue alters postural sway. Our aim was to better understand postural control mechanisms during upright stance following plantar flexor fatigue.

Method

Ten healthy young volunteers, 25.7 ± 2.2 years old, were recruited. Foot center-of-pressure (CoP) displacement data were collected during narrow base upright stance and eyes closed (i.e. blindfolded) conditions. Subjects were instructed to stand upright and as still as possible on a force platform under five test conditions: (1) non-fatigue standing on firm surface; (2) non-fatigue standing on foam; (3) ankle plantar flexor fatigue, standing on firm surface; (4) ankle plantar flexor fatigue, standing on foam; and (5) upper limb fatigue, standing on firm surface. An average of the ten 30-s trials in each of five test conditions was calculated to assess the mean differences between the trials. Traditional measures of postural stability and stabilogram-diffusion analysis (SDA) parameters were analyzed.

Results

Traditional center of pressure parameters were affected by plantar flexor fatigue, especially in the AP direction. For the SDA parameters, plantar flexor fatigue caused significantly higher short-term diffusion coefficients, and critical displacement in both mediolateral (ML) and anteroposterior (AP) directions. Long-term postural sway was different only in the AP direction.

Conclusions

Localized plantar flexor fatigue caused impairment to postural control mainly in the Sagittal plane. The findings indicate that postural corrections, on average, occurred at a higher threshold of sway during plantar flexor fatigue compared to non-fatigue conditions.     

MUTATIONS DO NOT ACCUMULATE IN ASEXUAL ISOLATES CAPABLE OF GROWTH AND EXTINCTION–MULLER'S RATCHET RE-EXAMINED

The rarity of exclusively asexual species is often attributed to Muller's Ratchet. This supposes that because asexual populations cannot recreate individuals with fewer mutations than the currently least-loaded line, mutations will accumulate in such isolates. However, because the computer models that corroborate this theory have assumed isolate immortality, it is possible that mutations will accumulate only if there is “soft” selection acting on relative, rather than absolute, fitness. Here we, therefore, describe several models in which 200 asexual organisms randomly selected from an infinite population in genetic equilibrium under “hard” selection (acting through absolute fitness), were followed for 100 generations. When there were no limits to the fluctuations in population size, the deterministic distribution of mutations per individual was maintained for 100 (as well as for 200) generations. If population growth was limited by a proportional decrease in fertility of the whole isolate, then the isolates tended to become extinct. The rate of extinction was inversely related to maximum isolate size. When resource limitation at maximum population size had an extra deleterious effect on mutants, then isolates shed the mutant classes. Mutations accumulated (ad inifinitum) in immortal isolates whose population numbers were kept constant by proportionately increasing or decreasing each class's size whenever isolate size ≠ 200. Muller's Ratchet, therefore, operates only when mutations affect the outcome of intraspecific contests, but not the organisms' intrinsic ability to survive in the ecosystem.