Macrophage migration inhibitory factor stimulated by Helicobacter pylori increases proliferation of gastric epithelial cells

AIM: Helicobacter pylori (H pylori) is associated with increased gastric inflammatory and epithelial expression of macrophage migration inhibitory factor (MIF) and gastric epithelial cell proliferation. This study aimed at determining whether H pylori directly stimulates release of MIF in monocytes, whether the cag pathogenicity island (PAI) is involved for this function, and whether MIF stimulated by H pylori increases gastric epithelial cell proliferation in vitro. METHODS: A cytotoxic wild-type H pylori strain (TN2)and its three isogenic mutants (TN2△cag, TN2△cagA and TN2△cagE) were co-cultured with cells of a human monocyte cell line, THP-1, for 24 h at different organism/cell ratios. MIF in the supernatants was measured by an ELISA. Cells of a human gastric cancer cell line, MKN45, were then co-cultured with the supernatants, with and without monoclonal anti-MIF antibody for 24 h. The cells were further incubated for 12 h after addition of (3)H-thymidine, and the levels of incorporation of (3)H-thymidine were measured with a liquid scintillation counter. RESULTS: The wild-type strain and the isogenic mutants, TN2△cagA and TN2△cagE, increased MIF release at organism/cell ratios of 200/1 and 400/1, but not at the ratios of 50/1 and 100/1. However, the mutant TN2△cag did not increase the release of MIF at any of the four ratios. (3)H-thymidine readings for MKN-45 cells were significantly increased with supernatants derived from the wild-type strain and the mutants TN2△cagA and TN2△cagE, but not from the mutant TN2△cag. Moreover, in the presence of monoclonal anti-MIF antibody, the stimulatory effects of the wild-type strain on cell proliferation disappeared. CONCLUSION: H pylori stimulates MIF release in monocytes, likely through its cag PAI, but not related to cagA or cagE. H pylori-stimulated monocyte culture supernatant increases gastric cell proliferation, which is blocked by anti-MIF antibody, suggesting that MIF plays an important role in H pylori-induced gastric epithelial cell proliferation

Insulator-to-metal transition in Kondo insulators under strong magnetic field

Magnetization curve and changes of the single-particle excitation spectra by magnetic field are calculated for the periodic Anderson model at half-filling in infinite spatial dimension by using the exact diagonalization method. It is found that the field-induced insulator-to-metal transition occurs at a critical field $H_c$, which is of the order of the single ion Kondo temperature. The transition is of first order, but could be of second order in the infinite system size limit. These results are compared with the experiments on the Kondo insulator YbB$_{12}$.Comment: 11 pages, REVTEX, no figures; 7 figures available on request; To appear in Phys. Rev. B, Mar.15, 199

Massless Thirring model in canonical quantization scheme

It is shown that the exact solvability of the massless Thirring model in the canonical quantization scheme originates from the intrinsic linearizability of its Heisenberg equations in the method of dynamical mappings. The corresponding role of inequivalent representations of free massless Dirac field is elucidated.Comment: 10 page

Stellar Rotation in Young Clusters. I. Evolution of Projected Rotational Velocity Distributions

Open clusters offer us the means to study stellar properties in samples with well-defined ages and initial chemical composition. Here we present a survey of projected rotational velocities for a large sample of mainly B-type stars in young clusters to study the time evolution of the rotational properties of massive stars. The survey is based upon moderate resolution spectra made with the WIYN 3.5 m and CTIO 4 m telescopes and Hydra multi-object spectrographs, and the target stars are members of 19 young open clusters with an age range of approximately 6 to 73 Myr. We made fits of the observed lines He I 4026, 4387, 4471 and Mg II 4481 using model theoretical profiles to find projected rotational velocities for a total of 496 OB stars. We find that there are fewer slow rotators among the cluster B-type stars relative to nearby B stars in the field. We present evidence consistent with the idea that the more massive B stars (M > 9 solar masses) spin down during their main sequence phase. However, we also find that the rotational velocity distribution appears to show an increase in the numbers of rapid rotators among clusters with ages of 10 Myr and higher. These rapid rotators appear to be distributed between the zero age and terminal age main sequence locations in the Hertzsprung-Russell diagram, and thus only a minority of them can be explained as the result of a spin up at the terminal age main sequence due to core contraction. We suggest instead that some of these rapid rotators may have been spun up through mass transfer in close binary systems.Comment: 33 pages, 11 figures, accepted by Ap

Induction of Nod2 in Myelomonocytic and Intestinal Epithelial Cells via Nuclear Factor-kB Activation

Nod2, a member of the Apaf1/Nod protein family, confers responsiveness to bacterial products and activates NF-kB, a ranscription factor that plays a central role in innate immunity. Recently, genetic variation in Nod2 has been associated with susceptibility to Crohn’s disease. Here, we report that expression of Nod2 is induced upon differentiation of CD34+ hematopoietic progenitor cells into granulocyte or monocyte/macrophages. In peripheral blood cells, the highest levels of Nod2 were observed in CD14+ (monocytes), CD15+ (granulocytes), and CD40+/CD86+ (dendritic cells) cell populations. Notably, stimulation of myeloblastic and epithelial cells with bacterial lipopolysaccharide or TNF resulted in up-regulation of Nod2. A search for consensus sites within the Nod2 promoter revealed a NF-kB binding element that was required for transcriptional activity in response to TNF . Moreover, ectopic expression of p65 induced transactivation, whereas that of dominant-negative I B blocked the transcriptional activity of the Nod2 promoter. Upon stimulation with TNF or lipopolysaccharide, both p50 and p65 subunits of NF-kB were bound to the Nod2 promoter. Thus, Nod2 expression is enhanced by proinflammatory cytokines and bacterial components via NF-kB, a mechanism that may contribute to the amplification of the innate immune response and susceptibility to inflammatory disease

Inhibition of HIF-1α by the anticancer drug TAS106 enhances X-ray-induced apoptosis in vitro and in vivo

In a previous study, we showed that a novel anticancer drug, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine (TAS106, ECyd) increased the antitumour efficacy of X-irradiation. However, its effects on hypoxic cells in tumours remain unclarified. Here, we show that TAS106 enhances the induction of apoptosis in X-irradiated human gastric adenocarcinoma MKN45 and MKN28 cells under hypoxia in vitro. At the same time, the accumulation of HIF-1α observed under hypoxia was shown to be decreased to the level of normoxia in the presence of 0.1 μM TAS106. To study the function of HIF-1α protein in apoptosis of hypoxic cells, we employed an HIF-1α reductive approach using its specific antisense oligodeoxynucleotide. The reduction of HIF-1α gene expression dramatically enhanced X-ray-induced apoptosis in hypoxic cells. In in vivo experiments in which MKN45 cells were transplanted into severe combined immunodeficient (SCID) mice, TAS106 (0.5 mg kg−1) suppressed HIF-1α expression and subsequently reduced the area of the hypoxic region in the tumour and enhanced the induction of apoptosis in the hypoxic region when combined with 2 Gy of X-irradiation. These results suggest the possibility that TAS106 acts as a potent radiosensitiser through the inhibition of HIF-1α expression and can be a useful agent against radiotherapy-resistant hypoxic cells in solid tumours

Realistic modeling of leakage and intrusion flows through leak openings in pipes

The hydraulics of leakage and intrusion flows through leak openings in pipes is complicated by variations in the leak areas owing to changes in pressure. This paper argues that the pressure–area relationship can reasonably be assumed to be a linear function, and a modified orifice equation is proposed for more realistic modeling of leakage and intrusion flows. The properties of the modified orifice equation are explored for different classes of leak openings. The implications for the current practice of using a power equation to model leakage and intrusion flows are then investigated. A mathematical proof is proposed for an equation linking the parameters of the modified orifice and power equations using the concept of a dimensionless leakage number. The leakage exponent of a given leak opening is shown to generally not be constant with variations in pressure and to approach infinity when the leakage number approaches a value of minus one. Significant modeling errors may result if the power equation is extrapolated beyond its calibration pressure range or at high exponent values. It is concluded that the modified orifice equation and leakage number provide a more realistic description of leakage and intrusion flows, and it is recommended that this approach be adopted in modeling studies

Stellar Rotation in Young Clusters. II. Evolution of Stellar Rotation and Surface Helium Abundance

We derive the effective temperatures and gravities of 461 OB stars in 19 young clusters by fitting the H-gamma profile in their spectra. We use synthetic model profiles for rotating stars to develop a method to estimate the polar gravity for these stars, which we argue is a useful indicator of their evolutionary status. We combine these results with projected rotational velocity measurements obtained in a previous paper on these same open clusters. We find that the more massive B-stars experience a spin down as predicted by the theories for the evolution of rotating stars. Furthermore, we find that the members of binary stars also experience a marked spin down with advanced evolutionary state due to tidal interactions. We also derive non-LTE-corrected helium abundances for most of the sample by fitting the He I 4026, 4387, 4471 lines. A large number of helium peculiar stars are found among cooler stars with Teff < 23000 K. The analysis of the high mass stars (8.5 solar masses < M < 16 solar masses) shows that the helium enrichment process progresses through the main sequence (MS) phase and is greater among the faster rotators. This discovery supports the theoretical claim that rotationally induced internal mixing is the main cause of surface chemical anomalies that appear during the MS phase. The lower mass stars appear to have slower rotation rates among the low gravity objects, and they have a large proportion of helium peculiar stars. We suggest that both properties are due to their youth. The low gravity stars are probably pre-main sequence objects that will spin up as they contract. These young objects very likely host a remnant magnetic field from their natal cloud, and these strong fields sculpt out surface regions with unusual chemical abundances.Comment: 50 pages 18 figures, accepted by Ap

Dose Dependent Effects on Cell Cycle Checkpoints and DNA Repair by Bendamustine

Bendamustine (BDM) is an active chemotherapeutic agent approved in the U. S. for treating chronic lymphocytic leukemia and non-Hodgkin lymphoma. Its chemical structure suggests it may have alkylator and anti-metabolite activities; however the precise mechanism of action is not well understood. Here we report the concentration-dependent effects of BDM on cell cycle, DNA damage, checkpoint response and cell death in HeLa cells. Low concentrations of BDM transiently arrested cells in G2, while a 4-fold higher concentration arrested cells in S phase. DNA damage at 50, but not 200 µM, was efficiently repaired after 48 h treatment, suggesting a difference in DNA repair efficiency at the two concentrations. Indeed, perturbing base-excision repair sensitized cells to lower concentrations of BDM. Timelapse studies of the checkpoint response to BDM showed that inhibiting Chk1 caused both the S- and G2-arrested cells to prematurely enter mitosis. However, whereas the cells arrested in G2 (low dose BDM) entered mitosis, segregated their chromosomes and divided normally, the S-phase arrested cells (high dose BDM) exhibited a highly aberrant mitosis, whereby EM images showed highly fragmented chromosomes. The vast majority of these cells died without ever exiting mitosis. Inhibiting the Chk1-dependent DNA damage checkpoint accelerated the time of killing by BDM. Our studies suggest that BDM may affect different biological processes depending on drug concentration. Sensitizing cells to killing by BDM can be achieved by inhibiting base-excision repair or disrupting the DNA damage checkpoint pathway

Nod2 Downregulates TLR2/1 Mediated IL1β Gene Expression in Mouse Peritoneal Macrophages

Nod2 is a cytosolic pattern recognition receptor. It has been implicated in many inflammatory conditions. Its signaling has been suggested to modulate TLR responses in a variety of ways, yet little is known about the mechanistic details of the process. We show in this study that Nod2 knockdown mouse peritoneal macrophages secrete more IL1β than normal macrophages when stimulated with peptidoglycan (PGN). Muramyl dipeptide (MDP, a Nod2 ligand) + PGN co-stimulated macrophages have lower expression of IL1β than PGN (TLR2/1 ligand) stimulated macrophages. MDP co-stimulation have similar effects on Pam3CSK4 (synthetic TLR2/1 ligand) mediated IL1β expression suggesting that MDP mediated down regulating effects are receptor dependent and ligand independent. MDP mediated down regulation was specific for TLR2/1 signaling as MDP does not affect LPS (TLR4 ligand) or zymosan A (TLR2/6 ligand) mediated IL1β expression. Mechanistically, MDP exerts its down regulating effects by lowering PGN/Pam3CSK4 mediated nuclear cRel levels. Lower nuclear cRel level were observed to be because of enhanced transporting back rather than reduced nuclear translocation of cRel in MDP + PGN stimulated macrophages. These results demonstrate that Nod2 and TLR2/1 signaling pathways are independent and do not interact at the level of MAPK or NF-κB activation