With Non-STelevation ACS, also low increases in troponin levels detected by highly sensitive assays were reported to be associated with a higher risk of cardiovascular death and myocardial infarction at 30 days and at 1 year. It is well known, that hs-cTnT is superior to MPO for rapid and accurate diagnosis of acute myocardial infarction among patients presenting with chest pain at the emergency department. Interestingly, MPO was not predictive for CE in patients with clinical TIMI risk score #3, whereas it was predictive in patients with higher risk scores. On the contrary, with increasing clinical TIMI risk scores, c-cTnT and hs-cTnT showed a gradual decline of the AUC for the prediction of CE within 30 days. Hence, risk prediction of biomarkers such as hs-cTnT and MPO clearly depends on the pretest probability. Further studies are needed to understand the different risk prediction profiles of hs-cTnT and MPO in low- and high-risk patients with suspected ACS. Interestingly, no improvement in risk prediction was observed with the combination of the clinical TIMI risk score and hs-cTnT with the pre-specified cut-off value of 13 pg/mL. Hence, in line with previous data, the recommended and arbitrary defined hs-cTnT decision limit seems to be less important for CE risk prediction than continuous hs-cTnT levels including also low-level increases. Furthermore, cut-off values of hs-cTnT may differ in various patient populations as has been suggested for other biomarkers such as NT-proBNP which shows dependency on age, gender, and body mass index. Limitations of this study are the single-centre design and the fact that risk assessment was only performed at time of presentation to the emergency department. However, this approach is in accordance with the original design of the TIMI risk score for prognostication at time of presentation. The rather high rate of patients presenting with ST-elevation myocardial infarction observed might at least in part be due to the fact that the study was performed at a tertiary referral center. However, the high rate of coronary angiographies associated with this constellation allowed for confirming or ruling out the diagnosis of coronary artery disease based on current gold standard. Nevertheless, the fact that decision making relied on c-cTnT measurements might have led to an underestimation of true ACS needing validation. As binary data whether events occurred or not were recorded in the study, time-to-event analyses could not be included. In conclusion, the combination of the patients’ clinical condition as represented in the clinical TIMI risk score, and a biomarker approach involving levels of continuous hs-cTnT, best predicted 30-day CE rate in Non-ST-elevation patients; thus, in this heterogeneous patient population the traditional but nevertheless sustainable clinical assessment remains fundamental for risk stratification.

Although we did not measure apoptosis or necrosis, these outcomes may influence the responses seen in these cell lines. There is clear evidence of a bystander effect in response to high doses of photons when we used the same serum and cell lines as for the neutron experiments. This result indicates that the methods used in our study are capable of detecting a bystander effect if such an effect exists. To the best of our knowledge there is no other factor that could have prevented neutrons from inducing a bystander effect in these cells, assuming a bystander effect even exists. Our findings are in agreement with previous studies that have reported the lack of a bystander effect on neutron exposure using clonogenic cell survival assay in a human skin cell line and zebrafish. However, other studies have reported contrasting results. Watson et al. found that transplantation of a mixture of neutron irradiated and unirradiated bone marrow cells into mice induced instability in the descendants of unirradiated cells as confirmed by measuring chromosomal aberrations, indicating that neutrons induce a bystander effect. However, since the gamma component in neutrons was 25%, it is possible that the observed bystander effect was due to the contaminating photons, which the authors did not rule out. Kinashi et al. studied a neutron-induced bystander effect in boron neutron capture therapy with a cell survival assay as well as cloning and sequencing methods. They reported an increase in the frequency of mutations in the hypoxanthine-guanine phosphoribosyltransferase locus in cells located near the irradiated cells. These results suggest that a neutron bystander effect may be comprised of gene mutations. The inability of fast neutrons to induce a cytogenetic bystander effect as shown here may be due to different types of damage induced at the molecular level compared to photons. Cellular recognition of DNA damage and the subsequent repair processes may differ between neutrons and photons. Furthermore, due to the lower levels of oxidative damage and free radical production by neutrons compared to photons, some of the critical bystander signaling pathways may not be activated. There is also the possibility that a neutron-induced bystander effect, if any, might depend on cell type, the endpoint being evaluated, and the energy of the neutrons. Neutrons, depending on their energy, might be more effective in controlling certain tumor types where conventional photon therapy is ineffective because the oxygen enhancement ratio, i.e. the differential radiosensitivity between poorly oxygenated and well-oxygenated cells, is reduced with neutrons. Unlike low-LET radiation, for high-LET radiation there is also a reduction in the differential radiosensitivity of cells related to their position in cell cycle. Recently, radiation-induced bystander cells were shown to rescue irradiated cells through intercellular feedback.

Genes encoding reverse transcriptase, calcium calmodulin-dependent protein kinase, replication protein and pre-mRNA-splicing factor were mapped to the Chr2 linkage group. These genes are important for essential cellular functions. The genes mapped to the Chr3 linkage group included those encoding guanine nucleotide exchange factor and cell division cycle-associated protein. Six different EST-SSRs associated with the gene encoding methionine-R-sulfoxide reductase were identified in the Chr4 linkage group. While some of these SSRs were located in close proximity to one-another, others were more widely separated. This may reflect the presence of large introns in this gene. A gene encoding anacetylcholinesterase was also mapped to this chromosome. Markers for the acetylcholinesterase gene and genes encoding zinc finger proteins were mapped to the Chr5 linkage group. Linkage group Chr6 was associated with genes encoding microtubule-associated protein and pyridoxine pyridoxamine 5phosphate oxidase. The brown planthopper genes encoding neuropeptide GPCR A5 and cytochrome P450 CYP6ER1 were mapped to linkage group Chr7, along with markers associated with EBNA2 binding protein p100 and ribosomal proteins. Linkage group Chr8 featured markers associated with the mucinlike protein gene, which may be important for the feeding behavior of brown planthoppers. Markers associated with histone RNA hairpin-binding protein, silencing protein and cysteine proteinase inhibitor precursor were mapped to the Chr9 linkage group. The Chr10 linkage group contained two markers for cytochrome P450 CYP6ER1, while the Chr11 linkage group contained 15 gene-specific annotated SSRs corresponding to the actin and chitin deacetylase genes. The EST-SSRs of the Chr12 linkage group were associated with chromodomain-helicase-DNAbinding protein and angiotensin converting protein. The Chr13 linkage group contained three EST-SSRs corresponding to the tyrosine-protein phosphatase gene. Only 3 gene-specific SSRs were anchored in the linkage group Chr14, corresponding to the gene for ESF1-like protein. Finally, 10 EST-SSRs corresponding to the vitellogenin gene were identified in the ChrX linkage group, along with markers for the transposase-like protein and deathassociated protein genes. We have constructed the most comprehensive linkage map for N. lugens that is currently available and demonstrated that brown planthopper virulence towards rice plants is controlled by a small number of genetic loci. To the best of our knowledge, this is the first study to successfully locate virulence factors in the genome of this important agricultural insect by marker-based genetic mapping. Building on the previous results, we constructed a high density molecular linkage map for the brown planthopper genome in order to enable the mapping of specific genes. The loci governing host preference and growth rate were delimited to specific regions flanked by molecular markers.

GLP-1 and GIP are generated within islets and exert possible unsuspected roles in the functional regulation of beta cells and other islet cell types. Their involvement in the regulation of beta cell mass is possibly more intriguing given the paucity of agents with such effects and the loss of beta cells in both type 1 and type 2 diabetes. Pregnancy is one of the very few situations associated with physiological and reversible expansion of beta cell mass not only in animals, which show remarkable plasticity of insulin secreting cells, but also in humans. Given the positive actions of the two incretins on beta cell mass, resulting from reciprocal effects on beta cell proliferation and death, we examined the role GLP-1 and GIP in islet adaptation to pregnancy using incretin receptor knockout mice. The results reveal an important role of GLP-1 in pregnancy-induced increases in beta cell mass, mediated largely by local GLP-1 production in alpha cells. In contrast, GIPR KO mice demonstrated intact mechanisms of islet adaptation to pregnancy, suggesting that islet or K-cell derived GIP is not essential for pregnancy-associated expansion of beta cell mass. Pregnancy increases metabolic needs and hence alters maternal metabolism by enhancing nutrient absorption and pancreatic beta cell function to match increased demand. Earlier reports on pregnancy-induced beta cell compensation revealed that lactogenic hormones stimulate beta cell proliferation and suppress apoptosis, thereby increasing beta cell mass. This is supported by observations in prolactin receptor knockout mice which displayed an inability to increase beta cell mass during pregnancy. Several reports also claim that lactogenic hormones increase serotonin biosynthesis in beta cells which in turn exerts proliferative effects on beta cells. In contrast, the involvement of gut hormones which normally play a key role in the regulation of beta cell function and survival has been largely overlooked. Interestingly, Sugiyama et al. observed that beta cell proliferation was not affected in pregnant glucagon-GFP knock-in mice lacking global proglucagon derived peptides, including glucagon, GLP-1, GLP-2 and oxyntomodulin. However, recent studies have shown that these mice display important compensatory changes even in the non-pregnant state, including markedly increased circulating GIP with substantial ectopic expression of GIP in islet beta cells. Thus, a positive role of GIP on beta cell mass may be particularly important in these animals. Accordingly, further studies are required using more specific receptor knockout models without functional GLP-1 or GIP receptors to assess the true role of incretin hormones in islet cell adaptations to pregnancy. Our study using GLP-1R and GIPR knockout animal models revealed that receptor knockout did not affect islet area and beta cell area but increased alpha cell area without affecting pancreatic glucagon content in GIP.

Indeed our observation that hydraulic permeability and cell size are correlated in untreated cells supports the notion that cell swelling increases hydraulic permeability, possibly by unfolding the cell membrane. In the presence of inflammatory stimulation, we theorize that inflammatory mediators act as triggers for disruption of the actin cytoskeleton, resulting in loss of cellular pre-stress and increased cell size. Increases in cell size due to inflammation may subsequently induce some unfolding of the cell membrane, resulting in increased hydraulic permeability without significant increases in Aqp-1 required. The cytoskeletal disruption findings and the loss of linear correlation between Lp and radius in inflammatory stimulated cells at both time points further confirm that these changes are irreversible in the current model system. Further studies are needed to determine if cell cytoskeletal changes are causative of changes in osmotic biophysical properties. The potential role of the cytoskeleton in the degenerative cascade associated with disc matrix catabolism, and the relationship between Aqp-1 and F-actin structure remain to be elucidated. A potential limitation of the current study is that the use of 2D culture may alter the NP cell phenotype, and may differentially regulate the inflammatory response. However, NPs are known to respond to cytokines both in vivo and in vitro, and our findings on biophysical responses are assessed on cells in 3D-like rounded morphology. Another limitation of the current study is that the NP cells were examined in an isolated, free-swelling environment. In vivo, these cells are surrounded by pericellular and extracellular matrices that confine the cells and contribute to cell volume regulation. Furthermore, these cells actively regulate their volume in vivo by increasing or decreasing the intracellular concentration of osmolytes. It has also been suggested that this process is dependent on the F-actin cytoskeleton, which we found to be disrupted by the inflammatory stimulation in the current study. The downstream consequences of altered cell mechano-sensitivity, including cell viability, metabolism, and ECM turnover, remain to be elucidated. Future studies will characterize the biophysical responses of NP cells to inflammatory stimulation in a 3D microenvironment that accounts for cell-matrix interactions. In summary, we found that the inflammatory stimuli LPS and TNF-a induced significant changes in the volume-response of NP cells to osmotic loading, including an increase in cell hydraulic permeability and cell size. These changes are associated with alterations in cytoskeletal structure in inflammatory-treated cells. Treated NP cells were unable to recover their biophysical properties to that of untreated cells after removal of inflammatory stimulant and return to basal media for 1 week.