A novel piggybac transposon inducible expression system identifies a role for akt signalling in primordial germ cell migration

In this work, we describe a single piggyBac transposon system containing both a tet-activator and a doxycycline-inducible expression cassette. We demonstrate that a gene product can be conditionally expressed from the integrated transposon and a second gene can be simultaneously targeted by a short hairpin RNA contained within the transposon, both in vivo and in mammalian and avian cell lines. We applied this system to stably modify chicken primordial germ cell (PGC) lines in vitro and induce a reporter gene at specific developmental stages after injection of the transposon-modified germ cells into chicken embryos. We used this vector to express a constitutively-active AKT molecule during PGC migration to the forming gonad. We found that PGC migration was retarded and cells could not colonise the forming gonad. Correct levels of AKT activation are thus essential for germ cell migration during early embryonic development

Number of siblings and the risk of solid tumours: a nation-wide study

We analysed the effects of number of siblings on the risk of solid tumours using the Swedish Family-Cancer Database, including population-based information on over 11 million individuals and more than 178 000 cancer patients diagnosed between 1958 and 2004. Incidence rate ratios (RRs), estimated by Poisson regression models, were adjusted for age, sex, birth cohort, area of residence and socioeconomic status. Having eight or more siblings vs none increased the risk of stomach cancer (RR=1.83, 95% confidence interval (CI), 1.44–2.34). Anal cancer diagnosed before age 40 showed the strongest association with the total siblings (RR=3.27, 95% CI, 2.04–5.26 for five or more siblings vs none). Endometrial (RR=0.76, 95% CI, 0.70–0.82), testicular (RR=0.71, 95% CI, 0.62–0.82), skin cancer (RR=0.82, 95% CI, 0.69–0.97) and melanoma (RR=0.72, 95% CI, 0.65–0.79) showed strong decreased risks for five or more siblings vs none. Prostate cancer risk for those with five or more older siblings vs none was 1.38 (95% CI, 1.23–1.55). Having five or more younger siblings was most strongly associated with stomach cancer (RR=1.59, 95% CI, 1.29–1.95) and melanoma (RR=0.68, 95% CI, 0.59–0.79). We conclude that sibship characteristics are strong correlates of cancer risk at several sites; plausible interpretations include socioeconomic status

Helicobacter pylori versus the Host: Remodeling of the Bacterial Outer Membrane Is Required for Survival in the Gastric Mucosa

Modification of bacterial surface structures, such as the lipid A portion of lipopolysaccharide (LPS), is used by many pathogenic bacteria to help evade the host innate immune response. Helicobacter pylori, a gram-negative bacterium capable of chronic colonization of the human stomach, modifies its lipid A by removal of phosphate groups from the 1- and 4′-positions of the lipid A backbone. In this study, we identify the enzyme responsible for dephosphorylation of the lipid A 4′-phosphate group in H. pylori, Jhp1487 (LpxF). To ascertain the role these modifications play in the pathogenesis of H. pylori, we created mutants in lpxE (1-phosphatase), lpxF (4′-phosphatase) and a double lpxE/F mutant. Analysis of lipid A isolated from lpxE and lpxF mutants revealed lipid A species with a 1 or 4′-phosphate group, respectively while the double lpxE/F mutant revealed a bis-phosphorylated lipid A. Mutants lacking lpxE, lpxF, or lpxE/F show a 16, 360 and 1020 fold increase in sensitivity to the cationic antimicrobial peptide polymyxin B, respectively. Moreover, a similar loss of resistance is seen against a variety of CAMPs found in the human body including LL37, β-defensin 2, and P-113. Using a fluorescent derivative of polymyxin we demonstrate that, unlike wild type bacteria, polymyxin readily associates with the lpxE/F mutant. Presumably, the increase in the negative charge of H. pylori LPS allows for binding of the peptide to the bacterial surface. Interestingly, the action of LpxE and LpxF was shown to decrease recognition of Helicobacter LPS by the innate immune receptor, Toll-like Receptor 4. Furthermore, lpxE/F mutants were unable to colonize the gastric mucosa of C57BL/6J and C57BL/6J tlr4 -/- mice when compared to wild type H. pylori. Our results demonstrate that dephosphorylation of the lipid A domain of H. pylori LPS by LpxE and LpxF is key to its ability to colonize a mammalian host

Host Responses in Life-History Traits and Tolerance to Virus Infection in Arabidopsis thaliana

Knowing how hosts respond to parasite infection is paramount in understanding the effects of parasites on host populations and hence host–parasite co-evolution. Modification of life-history traits in response to parasitism has received less attention than other defence strategies. Life-history theory predicts that parasitised hosts will increase reproductive effort and accelerate reproduction. However, empirical analyses of these predictions are few and mostly limited to animal-parasite systems. We have analysed life-history trait responses in 18 accessions of Arabidopsis thaliana infected at two different developmental stages with three strains of Cucumber mosaic virus (CMV). Accessions were divided into two groups according to allometric relationships; these groups differed also in their tolerance to CMV infection. Life-history trait modification upon virus infection depended on the host genotype and the stage at infection. While all accessions delayed flowering, only the more tolerant allometric group modified resource allocation to increase the production of reproductive structures and progeny, and reduced the length of reproductive period. Our results are in agreement with modifications of life-history traits reported for parasitised animals and with predictions from life-history theory. Thus, we provide empirical support for the general validity of theoretical predictions. In addition, this experimental approach allowed us to quantitatively estimate the genetic determinism of life-history trait plasticity and to evaluate the role of life-history trait modification in defence against parasites, two largely unexplored issues

Sex Determination:Why So Many Ways of Doing It?

Sexual reproduction is an ancient feature of life on earth, and the familiar X and Y chromosomes in humans and other model species have led to the impression that sex determination mechanisms are old and conserved. In fact, males and females are determined by diverse mechanisms that evolve rapidly in many taxa. Yet this diversity in primary sex-determining signals is coupled with conserved molecular pathways that trigger male or female development. Conflicting selection on different parts of the genome and on the two sexes may drive many of these transitions, but few systems with rapid turnover of sex determination mechanisms have been rigorously studied. Here we survey our current understanding of how and why sex determination evolves in animals and plants and identify important gaps in our knowledge that present exciting research opportunities to characterize the evolutionary forces and molecular pathways underlying the evolution of sex determination

Reinforcement effect of polyethylene fibre in root-filled teeth: comparison of two restoration techniques

Erdemir, Ali/0000-0002-6489-9620; yildirim, cihan/0000-0001-7218-9832; Erdemir, Ali/0000-0003-1140-3887WOS: 000235099100007PubMed: 16454794To compare in vitro two different fibre placement techniques on reinforcement of root-filled molar teeth with mesio-occluso-distal (MOD) cavities. Fifty sound extracted human mandibular molars were used (n = 10). Group 1 served as control. From group 2-5, endodontic access and standard MOD cavities were prepared. Following root canal treatment, group 2 was kept unrestored. In groups 3 and 4, the teeth were restored with composite resin (AP-X; Kuraray, Japan). A groove was prepared on occlusal surface of the finished restorations in group 4 from buccal to lingual direction. Polyethylene woven fibre (Ribbond; Seattle, WA, USA) was inserted in the groove in combination with flowable resin that was cured for 20 s and covered with composite resin. Cavity surfaces were covered with flowable resin in group 5, and polyethylene fibre was placed into the bed of resin in a buccal to lingual direction before the composite restoration was placed. All specimens were stored in 100% humidity at 37 degrees C for 24 h. Compressive loading of the teeth was performed by a universal testing machine at a crosshead speed of 0.5 mm min(-1) until failure. The data were recorded in Newton and submitted to ANOVA and Tukey post hoc test. The mean load necessary to fracture the samples in each group was: G1: 1671.57 +/- 131.54(a); G2: 375.21 +/- 34.30(b); G3: 749.47 +/- 124.54(c); G4: 1224.36 +/- 132.17(d); G5: 926.88 +/- 118.28(e). Different superscript letters demonstrate significant difference between groups. Polyethylene fibre use over or under MOD composite restorations significantly increased fracture strength. However, when the fibre was placed on the occlusal surface of the restoration in buccal to lingual direction, significantly higher fracture resistance was observed