Chironomus riparius (Diptera) genome sequencing reveals the impact of minisatellite transposable elements on population divergence

Active transposable elements (TEs) may result in divergent genomic insertion and abundance patterns among conspecific populations. Upon secondary contact, such divergent genetic backgrounds can theoretically give rise to classical Dobzhansky–Muller incompatibilities (DMI), thus contributing to the evolution of endogenous genetic barriers and eventually causing population divergence. We investigated differential TE abundance among conspecific populations of the nonbiting midge Chironomus riparius and evaluated their potential role in causing endogenous genetic incompatibilities between these populations. We focussed on a Chironomus‐specific TE, the minisatellite‐like Cla‐element, whose activity is associated with speciation in the genus. Using a newly generated and annotated draft genome for a genomic study with five natural C. riparius populations, we found highly population‐specific TE insertion patterns with many private insertions. A significant correlation of the pairwise F_ST estimated from genomewide single‐nucleotide polymorphisms (SNPs) and the F_ST estimated from TEs is consistent with drift as the major force driving TE population differentiation. However, the significantly higher Cla‐element F_ST level due to a high proportion of differentially fixed Cla‐element insertions also indicates selection against segregating (i.e. heterozygous) insertions. With reciprocal crossing experiments and fluorescent in situ hybridization of Cla‐elements to polytene chromosomes, we documented phenotypic effects on female fertility and chromosomal mispairings. We propose that the inferred negative selection on heterozygous Cla‐element insertions may cause endogenous genetic barriers and therefore acts as DMI among C. riparius populations. The intrinsic genomic turnover exerted by TEs may thus have a direct impact on population divergence that is operationally different from drift and local adaptation.     

An increase in intracellular Ca2+ is required for the activation of mitochondrial calpain to release AIF during cell death

Apoptosis-inducing factor (AIF), a flavoprotein with NADH oxidase activity anchored to the mitochondrial inner membrane, is known to be involved in complex I maintenance. During apoptosis, AIF can be released from mitochondria and translocate to the nucleus, where it participates in chromatin condensation and large-scale DNA fragmentation. The mechanism of AIF release is not fully understood. Here, we show that a prolonged ( approximately 10 min) increase in intracellular Ca(2+) level is a prerequisite step for AIF processing and release during cell death. In contrast, a transient ATP-induced Ca(2+) increase, followed by rapid normalization of the Ca(2+) level, was not sufficient to trigger the proteolysis of AIF. Hence, import of extracellular Ca(2+) into staurosporine-treated cells caused the activation of a calpain, located in the intermembrane space of mitochondria. The activated calpain, in turn, cleaved membrane-bound AIF, and the soluble fragment was released from the mitochondria upon outer membrane permeabilization through Bax/Bak-mediated pores or by the induction of Ca(2+)-dependent mitochondrial permeability transition. Inhibition of calpain, or chelation of Ca(2+), but not the suppression of caspase activity, prevented processing and release of AIF. Combined, these results provide novel insights into the mechanism of AIF release during cell death.     

Ubiquitin Is Covalently Attached to the p6Gag Proteins of Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus and to the p12Gag Protein of Moloney Murine Leukemia Virus

Host proteins are incorporated into retroviral virions during assembly and budding. We have examined three retroviruses, human immunodeficiency virus type 1 (HIV-1), simian immunodeficiency virus (SIV), and Moloney murine leukemia virus (Mo-MuLV), for the presence of ubiquitin inside each of these virions. After a protease treatment to remove exterior viral as well as contaminating cellular proteins, the proteins remaining inside the virion were analyzed. The results presented here show that all three virions incorporate ubiquitin molecules at approximately 10% of the level of Gag found in virions. In addition to free ubiquitin, covalent ubiquitin-Gag complexes were detected, isolated, and characterized from all three viruses. Our immunoblot and protein sequencing results on treated virions showed that approximately 2% of either HIV-1 or SIV p6^Gag was covalently attached to a single ubiquitin molecule inside the respective virions and that approximately 2 to 5% of the p12^Gag in Mo-MuLV virions was monoubiquitinated. These results show that ubiquitination of Gag is conserved among these retroviruses and occurs in the p6^Gag portion of the Gag polyprotein, a region that is likely to be involved in assembly and budding.     

Gender-based acute outcome in percutaneous coronary intervention of chronic total coronary occlusion

Background

Percutaneous coronary intervention (PCI) of total chronic coronary occlusion (CTO) still remains a major challenge. Insignificant data are reported in the literature about gender differences in CTO-PCI in the era of new drug-eluting stents. In this study we analysed the impact of gender on procedural characteristics, complications and acute results.

Methods

Between 2010–2015 we included 780 consecutive patients. They underwent PCI for at least one CTO. Antegrade and retrograde CTO techniques were applied.

Results

Patients undergoing CTO-PCI were mainly men (84%). Male patients were younger (66.9 years ±10.6 vs. 61.1 years ±10.4; p < 0.001), more often smokers, but less frequently had a history of coronary artery disease (24.4% vs. 32.7%; p = 0.085) compared with female patients. Female patients more often had diabetes mellitus (29.6% vs. 26.7%; p = 0.55) and hypertension (82.7% vs. 80.7%; p = 0.55). There were no differences with respect to the amount of contrast fluid, fluoroscopy time and examination time as well as to the length of the stent or the number of the stents. The stent diameter was slightly smaller in women, which was not surprising because the lumen calibre tends to be smaller in women than in men (3.0?mm (2.5–3) vs. 3.0?mm (3–3.5); p < 0.001). The success rates were 81.0% in women and 80.1% in men. There was no significant interaction between gender and procedural success and complication rates.

Conclusions

Our retrospective study suggests that women and men have a comparable success rate at a low complication rate after recanalisation of CTO.

     

The influence of L-ascorbic acid on the antibacterial-toxic activity of aflatoxins on adsorbent layer

AIMS: To substantiate the role of formaldehyde (HCHO) and its reaction products in the mechanism of the antibacterial-toxic effect of aflatoxins B1 (AFB1), B2, G1 and G2. MATERIALS AND RESULTS: Toxins were separated by overpressured layer chromatography, which was followed by biological evaluation directly on the adsorbent layer (BioArena system with Pseudomonas savastanoi pv. phaseolicola indicator bacteria). HCHO formed in this system was eliminated with exogenously added capturer molecule dimedone and L-ascorbic acid (AA) and measured as the adduct of dimedone and HCHO. The amount of HCHO was higher in the toxin-containing spots, particularly in the most toxic AFB1 spot, compared to a toxinless background. 0.1 mg ml(-1)AA augmented, 0.2 mg ml(-1) dimedone or 0.5 and 1 mg ml(-1) AA reduced the antibacterial effect of all four aflatoxins. CONCLUSION: The antibacterial-toxic effect of aflatoxins may be mediated by HCHO (and/or its reaction products) generated from bound HCHO forms in the bacterial cells. Basis of antibacterial-toxic activity of the four aflatoxins appears the same. SIGNIFICANCE AND IMPACT OF THE STUDY: Involvement of HCHO as a key molecule in the effect of aflatoxins indicates a totally new mechanism of action of these dangerous molecules. The BioArena system is useful to dissect the mode of action of antimicrobial compounds from different biological matrices.     

Fast cadmium inhibition of photosynthesis in cyanobacteria in vivo and in vitro studies using perturbed angular correlation of γ-rays

The effect of cadmium on the photosynthetic activity of Synechocystis PCC 6803 was monitored in this study. The oxygen evolving capacity of Synechocystis treated with 40 μM CdCl₂ was depressed to 10% of the maximum in 15 min, indicating that Cd²⁺ penetrated rapidly into the cells and blocked the photosynthetic activity. However, neither photosystem II (PSII) nor photosystem I (PSI) activity showed a significant short-term decrease which would explain this fast decrease in the whole-chain electron transport. Thermoluminescence measurements have shown that the charge separation and stabilization in PSII remains essentially unchanged during the first few hours following the Cd²⁺ treatment. The electron flow through PSI was monitored by following the redox changes of the P700 reaction centers of PSI. Alterations in the oxidation kinetics of P700 in the Cd²⁺-treated cells indicated that Cd²⁺ treatment might affect the available electron acceptor pool of P700, including the CO₂ reduction and accumulation in the cells. Perturbed angular correlation of γ-rays (PAC) using the radioactive ^111mCd isotope was used to follow the Cd²⁺ uptake at a molecular level. The most plausible interpretation of the PAC data is that Cd²⁺ is taken up by one or more Zn proteins replacing Zn²⁺ in Synechocystis PCC 6803. Using the radioactive ¹⁰⁹Cd isotope, a protein of approximately 30 kDa that binds Cd²⁺ could be observed in sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results indicate that Cd²⁺ might inactivate different metal-containing enzymes, including carbonic anhydrase, by replacing the zinc ion, which would explain the rapid and almost full inhibition of the photosynthetic activity in cyanobacteria.     

Strain-specific differences in pili formation and the interaction of <Emphasis Type="Italic">Corynebacterium diphtheriae</Emphasis> with host cells

Background  

Corynebacterium diphtheriae, the causative agent of diphtheria, is well-investigated in respect to toxin production, while little is known about C. diphtheriae factors crucial for colonization of the host. In this study, we investigated strain-specific differences in adhesion, invasion and intracellular survival and analyzed formation of pili in different isolates.     

Host plant-mediated variation in overwintering site quality: implications for the size and composition of populations of Acanthoscelides alboscutellatus (Coleoptera: Bruchidae)

This study provides an example of how variation in the quality of overwintering sites provided by the host plant of an insect seed predator can influence both the probability of overwintering survival and the size and composition of postwintering populations. Thus, the concept of host plant quality is extended to include variation in the suitability of the overwintering site of temperate region insects that overwinter within, or in habitats created by, their host plant. Adult Acanthoscelides alboscutellatus (Horn) (Coleoptera: Bruchidae) overwinter inside the fruit of Ludwigia alternifolia (L.) (Onagraceae). In early winter, however, fruits begin to dehisce, i.e., one or more of the fruit's four sides and/or top are shed. Variation in the onset and extent of dehiscence creates a range of overwintering habitats that vary in exposure to ambient conditions. In this study the frequency of possible overwintering sites in natural populations of L. alternifolia was determined by monitoring the phenology of fruit dehiscence from October through May in two populations for four years and for a third population for three years. Winter survivorship of adult A. alboscutellatus was assessed experimentally in eight environments representative of the conditions created by variation in dehiscence. These environments were produced by crossing four levels of exposure (degree of dehiscence) with two locations of the overwintering site, i.e., above or on the ground surface. The onset, phenology, and overall frequency of fruit dehiscence varied markedly among populations and years. Exposure, location, and their interaction had strong effects on survival and accounted for 80% of the observed variation in winter survival. Survivorship was higher on than above the ground, and in both locations decreased with increasing exposure. Thus, variation in fruit dehiscence among L. alternifolia populations will influence the size of postwintering A. alboscutellatus populations by dictating the quality of overwintering sites. Adult beetles that over-winter inside indehiscent fruit experience selection for small body size, associated with high mortality, when they attempt to exit the fruit at eclosion. As a consequence, the frequency of fruit dehiscence at eclosion coupled with the relative survival rates of adults within indehiscent fruit will determine the body size composition of postwintering populations and hence the response to selection for small body size in this species.