Which is the most prominent feature of the disease. Some candidates for these unknown ‘preeclampsia factors’ include pro-inflammatory cytokines, microparticles and anti-angiogenic factors. An imbalance between pro-angiogenic factors such as vascular endothelial growth factor and placental growth factor, and anti-angiogenic factors such as soluble fms-like tyrosine kinase 1 and soluble endoglin, has been observed before the onset of PE and after the clinical diagnosis. Endoglin, or CD105, is a homodimeric transmembrane glycoprotein localized on cell surfaces that functions as a coreceptor for transforming growth factor -b1 and TGF-b3 isoforms. Soluble endoglin might play an antiangiogenic effect in PE through binding to circulating TGF-b1, thus preventing its interaction with cell membrane, and consequently the pro-angiogenic and vasodilators effects of TGF-b1 in the normal endothelium. Previous studies demonstrated the involvement of TGF-b1 in the pathophysiology of PE but the results are conflicting. Some studies have found higher levels of TGF-b1 in PE, while in others there was no difference between PE and normotensive pregnancies. It is well established that an excessive maternal systemic inflammatory response to pregnancy is also involved in the pathogenesis of PE. Our research group has previously reported higher levels of pro-inflammatory cytokines, and decreased levels of the anti-inflammatory cytokine IL-10 in PE. Other studies have also reported that maternal circulating concentration of TNF soluble receptors sTNF-R1 and sTNF-R2 are significantly increased in preeclamptic and normotensive pregnant women under risk to develop this disease. The aim of this study was to evaluate the sEng, TGF-b1, sTNFR1 and sTNFR-2 levels in different forms of PE and to compare these variables to normotensive pregnant and healthy nonpregnant women, aiming to better understand the role of inflammation and endothelial dysfunction in PE pathogenesis. Our findings suggest that pregnancy is a condition associated with higher levels of anti-angiogenic and pro-inflammatory factors than the non-pregnant state and that PE is associated with an imbalance of these factors in the maternal circulation. In this study we compared the sEng, TGF-b1, sTNF-R1 and sTNFR-2 levels in women with different forms of PE, normotensive pregnant and healthy non-pregnant women. The past decades of research have brought a greater understanding of potential angiogenic imbalance that is likely involved in the PE pathophysiology. Eng is a pro-angiogenic agent that prevents apoptosis in hypoxic endothelial cells and is an essential component of the endothelial nitric oxide synthase activation complex, regulating nitric oxide-dependent regulation of vascular tone. In contrast, soluble Eng is an anti-angiogenic protein thought to impair TGF-b1 binding to its receptors and downstream signaling including effects on activation of eNOS and vasodilatation.

Many Gram-negative pathogens use N-acylhomoserine lactones as signaling molecules, whereas some Gram-positive bacteria use species-specific oligopeptides. A third cell-to-cell signal molecule is autoinducer-2, produced by a variety of Gram-negative and Gram-positive bacteria. AI-2 is therefore often called an interspecies signaling molecule. A few well-known pathogens, including Vibrio spp. and Salmonella, use AI-2 as a cue to sense population density. The key enzyme for AI-2 production is LuxS, which is an essential part of the activated methyl cycle, involved in recycling S-adenosylhomocysteine. More specifically, LuxS catalyzes the cleavage of S-ribosyl-homocysteine to homocysteine and 4,5-dihydroxy-2,3-pentanedione, which subsequently leads to the production of AI-2. A wide range of bacterial species produce AI-2, but evidence for the presence of signal reception and signal transduction pathways in organisms besides Escherichia coli, Vibrio spp. and Salmonella is lacking. While this lack of evidence is sometimes used to question the role of AI-2 in interspecies signaling, an alternative explanation is that other types of receptors and signal transduction pathways are yet to be discovered. Although the interspecies signaling molecule AI-2 is commonly linked to virulence and pathogenicity, it has recently been shown that the probiotic strain Lactobacillus acidophilus NCFM harbors a functional luxS gene and produces AI-2. Whether this signaling molecule plays a role in eliciting the beneficial traits of probiotic bacteria remains to be determined. Indeed, it was suggested that the ability to produce AI-2 affects attachment of L. acidophilus to intestinal epithelial cells, as a mutation in luxS was shown to result in decreased adherence to Caco-2 cells. Additionally, luxS has been attributed a central metabolic role in Lactobacillus reuteri 100–23 and Lactobacillus rhamnosus GG, and has been shown to influence adherence, biofilm formation and exopolysaccharide production in the latter. In a recent study AI-2 production has been demonstrated for three strains of Bifidobacteria and overexpression of luxS enhanced biofilm formation by Bifidobacterium longum NCC2705. In the present study we show that a functional luxS gene is widespread in the genus Bifidobacterium and that this gene in Bifidobacterium breve UCC2003 is involved in providing protection of Caenorhabditis elegans against Salmonella infection, a property which is linked to iron acquisition. Our data furthermore demonstrate that a functional luxS gene is required for murine gastrointestinal colonization by B. breve UCC2003. Members of the genus Bifidobacterium are recognized as being numerically dominant representatives of the microbiota of healthy breast-fed infants. Colonization of the newborn infant gut commences during and after birth by microbes from the mother and the environment.

Genome instability, and downstream activation of Notch and p53 pathways. Trace amounts of progerin have also been observed in several normal human tissues, although its biological significance and role in normal ageing remain to be determined. Antisense oligonucleotides can be designed to anneal to RNA by Watson-Crick hybridisation, and depending upon the base modifications and backbone chemistry, may exert their effects on gene expression through different mechanisms. An early application of AOs was to suppress expression of target gene and this was commonly achieved by recruitment of RNase H to degrade mRNA of a RNA: DNA oligonucleotide hybrid. AOs can also be used to redirect pre-mRNA processing. Since at least 74% of gene transcripts are alternatively spliced, splice-switching strategies could be broadly applicable to many different conditions. Furthermore, it is estimated that 10-15% of pathogenic mutations affect gene splicing, although this number is now considered to be an underestimate. AO induced exon skipping, exon retention and abrogation of the usage of alternative splice sites have been reported to by-pass or suppress pathogenic mutations in Duchenne muscular dystrophy, spinal muscular atrophy and thalassemia, respectively. Splice-switching AOs were able to mask abnormal splice sites in b-globin introns and force the aberrant splicing to default back to the normal pattern in b-thalassemia. Employing the same principle, abnormal LMNA splicing was suppressed by a phosphorodiamidate morpholino oligonucleotide annealed to the aberrant cryptic splice site in the LMNA exon 11 pre-mRNA in HGPS cells. Although splice-switching AOs can be used for therapeutic purposes by correcting defective gene transcripts, the same strategy can also be used to disrupt normal gene expression and induce pathological models of disease. Fong and colleagues have demonstrated the activation of the cryptic splice site activated in HGPS in normal human fibroblasts by targeting 29-O-methoxyethyl AOs to motifs near the cryptic splice site in exon 11. Ageing in skeletal muscle is associated with loss of muscle bulk and strength, eventually resulting in significant functional disabilities. The processes responsible for muscle senescence are incompletely understood, but it is known that multiple factors play a role, including the accumulation of lifelong exposure to extrinsic detrimental factors like exercise damage, accumulative mitochondrial DNA mutations, increased free radicals and decreased oxidative response, reduced protein turnover capacity, low-grade systemic inflammation, and impaired neuromuscular junction function. Nevertheless, there are few models that specifically address muscle ageing. In another study, we observed low-level accumulation of progerin in normal human skeletal muscle, but it is unclear if the levels detected are sufficient to play a role in the ageing process.

We first investigated the ESE potential of the CAAACAA sequence found in pig Smn1 exon 7 to evaluate the feasibility of constructing a pig Smn2-like model by introducing a single +6C.T mutation in the endogenous porcine Smn1 gene. We introduced the ESE region into the pSXN13 splicing reporter minigene and observed minimal inclusion of the alternative exon in the wild type construct and complete loss of inclusion when we introduced a C.T mutation analogous to the + 6T in SMN2. This demonstrates splicing enhancer activity of the altered pig ESE and further demonstrates that a C.T mutation can abrogate this activity despite the lack of an AG dinucleotide within the ESE which has previously been proposed to form an hnRNP A1 ESS in SMN2 exon 7. Therefore, in this context the pig ESE supports the ESE-loss model. When we disrupted the upstream ESS motif by an A.C mutation, the splicing pattern of the wild type pig ESE and mutant pig ESE was very similar to that of SMN1 and SMN2. However, the inclusion level of the wild type pig Smn1-like ESE construct seemed slightly lower than that of the wild type SMN1 construct, while the inclusion level of the mutant pig Smn2-like ESE construct seemed slightly higher than that of the SMN2 construct. These results can be explained by the ESE-loss/ ESS-gain model. The G.A change in the pig ESE seems to have decreased the ESE activity relative to the human SMN1 ESE sequence, but in the context of the +6C.T mutation, which completely abolishes the human SMN1 ESE activity, the activity of the gained ESS has also been decreased. Overall, the regulatory element is more neutral in the pig Smn1 gene than in human SMN1 and SMN2, although it retains some ESE activity. When we introduced pig Smn1 exon 7 with flanking regions in our SMN model minigene, we observed only a modest decrease in exon inclusion between the construct with the wild type sequence and the +6C.T mutant construct, indicating that in the native pig Smn1 gene a functional ESE is not crucial for exon 7 inclusion. We then examined pig intron 7 and found that in the region of the previously reported IVS7-ISS, there were differences between the human and pig sequence which could potentially alter binding affinity of one or both of the hnRNP A1 sites contained within the ISS motif. In fact, the motif score was decreased for the proximal hnRNP A1 site and increased for the distal site through an A.C mutation which has previously been shown to increase skipping of human SMN2 exon 7. These findings then lead us to investigate the ISS activity of the downstream region in pig Smn1 intron 7 and we found that when we inserted the human ISS into the pig context, a splicing pattern more similar to SMN2 splicing pattern was observed when we introduced the +6C.T mutation into the pig ESE sequence. When we introduced mutations previously reported to remove ISS activity, we observed exon inclusion similar to the wild type pig construct.

Among multiple potential source populations, and their maximum assignment probability is more likely to fall below the threshold. Thus, the contributions of reporting groups that are less distinct can be under estimated in the mixture. Whether this bias affects estimates of population-specific traits depends on whether the trait is negatively or positively correlated with genetic similarity among reporting groups. In the positive case, trait estimates from IA are expected to be unbiased. Even though we might underestimate the true number of fish from genetically similar reporting groups, we are unlikely to end up with a biased set of individuals relative to the traits they express. Precision might suffer from diminished sample size, but trait estimates should not be biased in the same way that population mixture composition can be. Although bias was not necessarily expected in comparing IA and BMM, our results serve to underscore a classical problem in biological sampling – the need to balance accuracy of IA with sample size and therefore statistical power. On one hand, IA with a high MAP-rule threshold will better assure that individuals included in the analysis are accurately classified. On the other hand, if mixture samples are small to begin with, as they often are in ecological studies, then the number of individuals with assignment probabilities above the MAP threshold can be quite small. If one lowers the threshold to include more individuals, assignment error and therefore trait estimation bias can increase. This is not such a problem if the trait distribution is correlated with the genetic structure of reporting groups, but if genetically similar reporting groups have very different trait values, then clearly misassignments will bias estimates of population-specific traits. We saw signs of such bias in our simulation studies where we created trait distributions having both positive and negative correlation with genetic distance. The genetic data we used were based on a regional subset of the GAPS-Chinook microsatellite baseline. The 13 GAPS loci are highly variable, with nearly 500 alleles coast wide and highly significant allele frequency differences among populations and among regions. This diversity provided significant power to accurately assign individual Chinook salmon to their putative population of origin. The difference in performance between IA and BMM would be greater in applications with less powerful baselines, which could be caused by fewer, less variable loci, low differentiation among populations, or because mixture samples are degraded and provide quality genotypes for only a subset of loci. In many ecological genetic studies of the type we consider here, power is often limited by small mixture samples. Genetic reporting groups that have small contributions to a mixture may provide very few individuals from which to estimate trait parameters.