Epizootic Hemorrhagic Disease Virus Induces and Benefits from Cell Stress,Autophagy, and Apoptosis

The mode and timing of virally induced cell death hold the potential of regulating viral yield, viral transmission, and the severity of virally induced disease. Orbiviruses such as the epizootic hemorrhagic disease virus (EHDV) are nonenveloped and cytolytic. To date, the death of cells infected with EHDV, the signal transduction pathways involved in this process, and the consequence of their inhibition have yet to be characterized. Here, we report that the Ibaraki strain of EHDV2 (EHDV2-IBA) induces apoptosis, autophagy, a decrease in cellular protein synthesis, the activation of c-Jun N-terminal kinase (JNK), and the phosphorylation of the JNK substrate c-Jun. The production of infectious virions decreased upon inhibition of apoptosis with the pan-caspase inhibitor Q-VD-OPH (quinolyl-valyl-O-methylaspartyl-[-2,6-difluorophenoxy]-methyl ketone), upon inhibition of autophagy with 3-methyladenine or via the knockout of the autophagy regulator Atg5, or upon treatment of infected cells with the JNK inhibitor SP600125 or the cyclin-dependent kinase (CDK) inhibitor roscovitine, which also inhibited c-Jun phosphorylation. Moreover, Q-VD-OPH, SP600125, and roscovitine partially reduced EHDV2-IBA-induced cell death, and roscovitine diminished the induction of autophagy by EHDV2-IBA. Taken together, our results imply that EHDV induces and benefits from the activation of signaling pathways involved in cell stress and death.     

Susceptibility and physiological responses of Jatropha curcas accessions to broad mite infestation

The broad mite Polyphagotarsonemus latus is a key pest of physic nut (Jatropha curcas L.). The purpose of this study was to identify physic nut accessions that are less susceptible to P. latus, in support of the breeding program of J. curcas. We first evaluated population growth rate and injury symptoms of P. latus on different J. curcas accessions and then carried out physiological analyses on P. latus-infested and uninfested accessions. From the germplasm bank of the Federal University of Viçosa, 15 physic nut accessions with high seed oil content, with different genetic background, were tested. The following traits were evaluated: instantaneous population growth rate of P. latus (r _i ), injury symptoms, relative leaf water content, specific leaf area, gas exchange, photosynthetic pigments, nitrogen and biomass of the aerial part. Significant differences were observed for P. latus population growth rate and injury symptoms among accessions. A positive correlation between P. latus growth rate and injury was found. The UFVJC72 accession stood out as the more resistant, considering P. latus growth rate and injury symptoms, compared with most accessions. Physiological responses did not vary among accessions, but did between infested and uninfested plants. In P. latus-infested plants, net photosynthesis was on average 50.5 % lower than in uninfested plants, whereas stomatal conductance and transpiration decreased by 46.2 and 51.6 %, respectively.     

Comparison between mechanical stress and bone mineral density in the femur after total hip arthroplasty by using subject-specific finite element analyses

The mechanism underling bone mineral density (BMD) loss that occurs in the femur after total hip arthroplasty (THA) remains unknown. We compared the equivalent stress and strain energy density (SED) to BMD in the femur after THA using subject-specific finite element analyses. Twenty-four patients who had undergone primary cementless THA were analysed. BMD was measured using dual-energy X-ray absorptiometry (DEXA) at 1 week and 3, 6 and 12 months after THA. Seven regions of interest (ROIs) were defined in accordance with Gruen's system (ROIs 1–7). Computed tomography images of the femurs were acquired pre- and postoperatively, and the images were converted into three-dimensional finite element (FE) models. Equivalent stress and SED were analysed and compared with DEXA data. BMD was maintained 1 year after THA in ROIs 3, 4, 5 and 6, whereas BMD decreased in ROIs 1, 2 and 7. FE analysis revealed that equivalent stress in ROIs 3, 4, 5 and 6 was much higher than that in ROIs 1, 2 and 7. A significant correlation was observed between the rate of changes in BMD and equivalent stress. Reduction of equivalent stress may contribute to decrease in BMD in the femur after THA.     

Raymond Gosling: the man who crystallized genes

On April 25th 1953, three publications in Nature forever changed the face of the life sciences in reporting the structure of DNA. Sixty years later, Raymond Gosling shares his memories of the race to the double helix.It has not escaped our attention that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material.James D Watson and Francis HC Crick [1]By including this statement in their April 25th 1953 Nature article describing a model for the structure of DNA, Watson and Crick made one of the great understatements in history. In that moment, the seeds for the double helix''s infamy - alongside the names ''Watson'' and Crick'' - were sown. Lesser known outside scientific circles is that this article did not include one iota of experimental data: Watson and Crick, who were based at the University of Cambridge''s Cavendish laboratory, contributed deductive reasoning alone to the double helix model, albeit reasoning of an undoubtedly Nobel Prize-worthy standard. Instead, as has now been described many times, the model relied on X-ray diffraction data obtained by others, at King''s College, and these data did not reach Watson and Crick by entirely wholesome means [2]. To add to the insult, Watson and Crick''s report of the double helix did not fully credit the work of King''s as being essential to the construction of their model, although the King''s team did enjoy co-publication of their data alongside the double helix article, in the form of two articles in the same issue of Nature [3,4]. One of these articles described the X-ray diffraction work performed by senior researcher Rosalind E Franklin, together with PhD student Raymond G Gosling, and contained the highest quality diffraction patterns yet achieved for DNA [3]. It was these data that had proved invaluable in Watson and Crick''s quest for the double helix.Earlier still, before Franklin arrived at King''s, Gosling had achieved a major breakthrough in the search for DNA''s structure when he became the first person to crystallize genes, under the guidance of Maurice Wilkins, who was the lead author of the other King''s article to accompany Watson and Crick''s model [4].Watson published his controversial memoir of the discovery, aptly named ''The Double Helix'', in the 1960s [5], and in doing so propelled the story to worldwide fame, establishing DNA''s structure as an icon of science in the popular imagination. However, events were relayed in Watson''s book very much from his own point of view and at times, it has been argued, even verged on the fictitious.Aside from Watson, Ray Gosling is the only surviving member of the select group of seven scientists to feature as an author on one of the three Nature articles. Gosling and his wife, Mary, were kind enough to welcome Genome Biology into their home, where he shared with us his perspective of the events of 60 years ago.Elsewhere, Genome Biology has marked the anniversary by canvassing our Editorial Board for their opinions on the most important advances in the field since 1953 [6].     

Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin

Sperm cryopreservation is useful for the effective storage of genomic resources derived from genetically engineered mice. However, freezing the sperm of C57BL/6 mice, the most commonly used genetic background for genetically engineered mice, considerably reduces its fertility. We previously reported that methyl-beta-cyclodextrin dramatically improved the fertility of frozen/thawed C57BL/6 mouse sperm. Recently, it was reported that exposing sperm to reduced glutathione may alleviate oxidative stress in frozen/thawed mouse sperm, thereby enhancing in vitro fertilization (IVF); however, the mechanism underlying this effect is poorly understood. In the present study, we examined the combined effects of methyl-beta-cyclodextrin and reduced glutathione on the fertilization rate of IVF with frozen/thawed C57BL/6 mouse sperm and the characteristic changes in the zona pellucida induced by reduced glutathione. Adding reduced glutathione to the fertilization medium increased the fertilization rate. Methyl-beta-cyclodextrin and reduced glutathione independently increased fertilization rates, and their combination produced the strongest effect. We found that reduced glutathione increased the amount of free thiols in the zona pellucida and promoted zona pellucida enlargement. Finally, 2-cell embryos produced by IVF with the addition of reduced glutathione developed normally and produced live offspring. In summary, we have established a novel IVF method using methyl-beta-cyclodextrin during sperm preincubation and reduced glutathione during the IVF procedure to enhance fertility of frozen/thawed C57BL/6 mouse sperm.     

Let the right one in: a microeconomic approach to partner choice in mutualisms

One of the main problems impeding the evolution of cooperation is partner choice. When information is asymmetric (the quality of a potential partner is known only to himself), it may seem that partner choice is not possible without signaling. Many mutualisms, however, exist without signaling, and the mechanisms by which hosts might select the right partners are unclear. Here we propose a general mechanism of partner choice, "screening," that is similar to the economic theory of mechanism design. Imposing the appropriate costs and rewards may induce the informed individuals to screen themselves according to their types and therefore allow a noninformed individual to establish associations with the correct partners in the absence of signaling. Several types of biological symbioses are good candidates for screening, including bobtail squid, ant-plants, gut microbiomes, and many animal and plant species that produce reactive oxygen species. We describe a series of diagnostic tests for screening. Screening games can apply to the cases where by-products, partner fidelity feedback, or host sanctions do not apply, therefore explaining the evolution of mutualism in systems where it is impossible for potential symbionts to signal their cooperativeness beforehand and where the host does not punish symbiont misbehavior.     

ZouA, a putative relaxase, is essential for dna amplification in Streptomyces kanamyceticus

Previously, we showed that a 145-kb DNA region, including the entire kanamycin biosynthetic gene cluster (with two kanamycin resistance genes), was tandemly amplified up to 36-fold in an industrial strain of Streptomyces kanamyceticus. Strain improvement had included the use of increased kanamycin resistance as an initial potential indicator of higher kanamycin productivity. We were able to recapitulate the DNA amplification by cultivating S. kanamyceticus under selection for kanamycin resistance. To identify the genes required for amplification, various chromosome deletions were constructed, and the DNA amplification was shown to depend on orf1082 (zouA), present in a putative mobile genetic element. ZouA consists of 1,481 amino acids and is homologous to the products of traA-like genes of some conjugative plasmids. These genes encode relaxases that initiate DNA transfer during conjugation by single-strand nicking at oriT. As in the original high-producing strain, DNA amplification occurred between 16-nucleotide (nt) sites (RsA and RsB) containing 14 identical nucleotides. Interestingly, RsA lies just 80 bp upstream of the initiation codon of zouA and is partially contained in an inverted repeat structure similar to those found in plasmid oriT sequences, suggesting that it might function in a manner similar to that of oriT. We therefore propose that DNA amplification in S. kanamyceticus is initiated by relaxase-mediated recombination between oriT-related sequences.