Gα₁₂ gep oncogene deregulation of p53-responsive microRNAs promotes epithelial–mesenchymal transition of hepatocellular carcinoma

Yang, Y M,Lee, W H,Lee, C G,An, J,Kim, E-S,Kim, S H,Lee, S-K,Lee, C H,Dhanasekaran, D N,Moon, A,Hwang, S,Lee, S J,Park, J-W,Kim, K M,Kim, S G Macmillan Publishers Limited 2015 Oncogene Vol.34 No.22

Hepatocellular carcinoma (HCC) has a poor prognosis owing to aggressive phenotype. Gα<SUB>12</SUB> gep oncogene product couples to G-protein-coupled receptors, whose ligand levels are frequently increased in tumor microenvironments. Here, we report Gα<SUB>12</SUB> overexpression in human HCC and the resultant induction of zinc-finger E-box-binding homeobox 1 (ZEB1) as mediated by microRNA deregulation. Gα<SUB>12</SUB> expression was higher in HCC than surrounding non-tumorous tissue. Transfection of Huh7 cell with an activated mutant of Gα<SUB>12</SUB> (Gα<SUB>12</SUB>QL) deregulated microRNA (miRNA or miR)-200b/a/429, -194-2/192 and -194-1/215 clusters in the miRNome. cDNA microarray analyses disclosed the targets affected by Gα<SUB>12</SUB> gene knockout. An integrative network of miRNAs and mRNA changes enabled us to predict ZEB1 as a key molecule governed by Gα<SUB>12</SUB>. Decreases of miR-200a/b, -192 and -215 by Gα<SUB>12</SUB> caused ZEB1 induction. The ability of Gα<SUB>12</SUB> to decrease p53 levels, as a result of activating protein-1 (AP-1)/c-Jun-mediated mouse double minute 2 homolog induction, contributed to transcriptional deregulation of the miRNAs. Gα<SUB>12</SUB>QL induced ZEB1 and other epithelial–mesenchymal transition markers with fibroblastoid phenotype change. Consistently, transfection with miR-200b, -192 or -215 mimic prevented the ability of Gα<SUB>12</SUB>QL to increase tumor cell migration/invasion. In xenograft studies, sustained knockdown of Gα<SUB>12</SUB> decreased the overall growth rate and average volume of tumors derived from SK-Hep1 cell (mesenchymal-typed). In HCC patients, miR-192, -215 and/or -200a were deregulated with microvascular invasion or growth advantage. In the HCC samples with higher Gα<SUB>12</SUB> level, a correlation existed in the comparison of relative changes of Gα<SUB>12</SUB> and ZEB1. In conclusion, Gα<SUB>12</SUB> overexpressed in HCC causes ZEB1 induction by deregulating p53-responsive miRNAs, which may facilitate epithelial–mesenchymal transition and growth of liver tumor. These findings highlight the significance of Gα<SUB>12</SUB> upregulation in liver tumor progression, implicating Gα<SUB>12</SUB> as an attractive therapeutic target.

A POSSIBLE BINARY SYSTEM OF A STELLAR REMNANT IN THE HIGH-MAGNIFICATION GRAVITATIONAL MICROLENSING EVENT OGLE-2007-BLG-514

Miyake, N.,Udalski, A.,Sumi, T.,Bennett, D. P.,Dong, S.,Street, R. A.,Greenhill, J.,Bond, I. A.,Gould, A.,Kubiak, M.,Szymań,ski, M. K.,Pietrzyń,ski, G.,Soszyń,ski, I.,Ulaczyk, K.,Wyrzyk IOP Publishing 2012 The Astrophysical journal Vol.752 No.2

<P>We report the extremely high-magnification (A > 1000) binary microlensing event OGLE-2007-BLG-514. We obtained good coverage around the double peak structure in the light curve via follow-up observations from different observatories. The binary lens model that includes the effects of parallax (known orbital motion of the Earth) and orbital motion of the lens yields a binary lens mass ratio of q = 0.321 +/- 0.007 and a projected separation of s = 0.072 +/- 0.001 in units of the Einstein radius. The parallax parameters allow us to determine the lens distance D-L = 3.11 +/- 0.39 kpc and total mass M-L = 1.40 +/- 0.18 M-circle dot; this leads to the primary and secondary components having masses of M-1 = 1.06 +/- 0.13 M-circle dot and M-2 = 0.34 +/- 0.04 M-circle dot, respectively. The parallax model indicates that the binary lens system is likely constructed by the main-sequence stars. On the other hand, we used a Bayesian analysis to estimate probability distributions by the model that includes the effects of xallarap (possible orbital motion of the source around a companion) and parallax (q = 0.270 +/- 0.005, s = 0.083 +/- 0.001). The primary component of the binary lens is relatively massive, with M-1 = 0.9(-0.3)(+4.6) M-circle dot and it is at a distance of D-L = 2.6(-0.9)(+3.8) kpc. Given the secure mass ratio measurement, the companion mass is therefore M-2 = 0.2(-0.1)(+1.2) M-circle dot. The xallarap model implies that the primary lens is likely a stellar remnant, such as a white dwarf, a neutron star, or a black hole.</P>

OGLE-2011-BLG-0265Lb: A JOVIAN MICROLENSING PLANET ORBITING AN M DWARF

Skowron, J.,Shin, I.-G.,Udalski, A.,Han, C.,Sumi, T.,Shvartzvald, Y.,Gould, A.,Dominis Prester, D.,Street, R. A.,Jørgensen, U. G.,Bennett, D. P.,Bozza, V.,Szymań,ski, M. K.,Kubiak, M.,Pietrzy
IOP Publishing 2015 The Astrophysical journal Vol.804 No.1

<P>We report the discovery of a Jupiter-mass planet orbiting an M-dwarf star that gave rise to the microlensing event OGLE-2011-BLG-0265. Such a system is very rare among known planetary systems and thus the discovery is important for theoretical studies of planetary formation and evolution. High-cadence temporal coverage of the planetary signal, combined with extended observations throughout the event, allows us to accurately model the observed light curve. However, the final microlensing solution remains degenerate, yielding two possible configurations of the planet and the host star. In the case of the preferred solution, the mass of the planet is M-p = 0.9 +/- 0.3 M-J, and the planet is orbiting a star with a mass M = 0.22 +/- 0.06 M-circle dot. The second possible configuration (2 sigma away) consists of a planet with M-p = 0.6 +/- 0.3M(J) and host star with M = 0.14 +/- 0.06M(circle dot). The system is located in the Galactic disk 3-4 kpc toward the Galactic bulge. In both cases, with an orbit size of 1.5-2.0 AU, the planet is a 'cold Jupiter'-located well beyond the 'snow line' of the host star. Currently available data make the secure selection of the correct solution difficult, but there are prospects for lifting the degeneracy with additional follow-up observations in the future, when the lens and source star separate.</P>

Restricted growth of U‐type infectious haematopoietic necrosis virus (IHNV) in rainbow trout cells may be linked to casein kinase II activity

Park, J W,Moon, C H,Harmache, A,Wargo, A R,Purcell, M K,Bremont, M,Kurath, G Blackwell Publishing Ltd 2011 Journal of fish diseases Vol.34 No.2

<P><B>Abstract</B></P><P>Previously, we demonstrated that a representative M genogroup type strain of infectious haematopoietic necrosis virus (IHNV) from rainbow trout grows well in rainbow trout‐derived RTG‐2 cells, but a U genogroup type strain from sockeye salmon has restricted growth, associated with reduced genome replication and mRNA transcription. Here, we analysed further the mechanisms for this growth restriction of U‐type IHNV in RTG‐2 cells, using strategies that assessed differences in viral genes, host immune regulation and phosphorylation. To determine whether the viral glycoprotein (G) or non‐virion (NV) protein was responsible for the growth restriction, four recombinant IHNV viruses were generated in which the G gene of an infectious IHNV clone was replaced by the G gene of U‐ or M‐type IHNV and the NV gene was replaced by NV of U‐ or M‐type IHNV. There was no significant difference in the growth of these recombinants in RTG‐2 cells, indicating that G and NV proteins are not major factors responsible for the differential growth of the U‐ and M‐type strains. Poly I:C pretreatment of RTG‐2 cells suppressed the growth of both U‐ and M‐type IHNV, although the M virus continued to replicate at a reduced level. Both viruses induced type 1 interferon (IFN1) and the IFN1 stimulated gene Mx1, but the expression levels in M‐infected cells were significantly higher than in U‐infected cells and an inhibitor of the IFN1‐inducible protein kinase PKR, 2‐aminopurine (2‐AP), did not affect the growth of U‐ or M‐type IHNV in RTG‐2 cells. These data did not indicate a role for the IFN1 system in the restricted growth of U‐type IHNV in RTG‐2 cells. Prediction of kinase‐specific phosphorylation sites in the viral phosphoprotein (P) using the NetPhosK program revealed differences between U‐ and M‐type P genes at five phosphorylation sites. Pretreatment of RTG‐2 cells with a PKC inhibitor or a p38MAPK inhibitor did not affect the growth of the U‐ and M‐type viruses. However, 100 μ<SMALL>m</SMALL> of the casein kinase II (CKII) inhibitor, 5,6‐dichloro‐1‐β‐<SMALL>d</SMALL>‐ribofuranosylbenzimidazole (DRB), reduced the titre of the U type 8.3‐fold at 24 h post‐infection. In contrast, 100 μ<SMALL>m</SMALL> of the CKII inhibitor reduced the titre of the M type only 1.3‐fold at 48 h post‐infection. Our data suggest that the different growth of U‐ and M‐type IHNV in RTG‐2 cells may be linked to a differential requirement for cellular protein kinases such as CKII for their growth.</P>

Indirect Method for Quantification of Cellular Biomass in a Solidscontaining Medium Used as Pre-culture for Cellulase Production

F. M. Cunha,A. L. G. Bacchin,A. C. L. Horta,T. C. Zangirolami,A. C. Badino,C. S. Farinas 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.1

A process that combines the advantages of solid state fermentation (SSF) and submerged fermentation (SmF) could increase the efficiency of cellulase production required in the cellulosic ethanol industry. Due to the difficulty of measuring cellular biomass in the presence of solids, we developed a novel methodology for indirect quantification of biomass during production of the preculture for a combined fermentation process. Cultivation of Aspergillus niger was initiated as SSF using sugar cane bagasse as a solid substrate. Experiments were conducted in the absence of bagasse to determine growth kinetic parameters. Changes in glucose and biomass concentrations were measured. and the data were used for simulation employing a simple unstructured model. Parameters were estimated by applying a combination of Simulated Annealing (SA) and Levenberg-Marquardt (LM) algorithms to search for minimization of the error between model estimates and experimental data. Growth kinetics followed the Contois model, with a maximum specific growth rate (μmax) of 0.042/h, a yield coefficient for biomass formation (Yx/s) of 0.30 g/g and a death constant (kD) of 0.005/h.These parameters were used to simulate cellular growth in the solids-containing medium. The proposed model accurately described the experimental data and succeeded in simulating the cell concentration profile. The selected pre-culture conditions (24 h as SSF followed by 48 h as SmF) were applied for cellulase production using the combined fermentation process and resulted in an endoglucanase activity (1,052 ± 34 U/L) greater than that obtained using the conventional SmF procedure (824 ± 44 U/L). Besides the standardization of pre-culture conditions, this methodology could be very useful in systems where direct measurement of cell mass is not possible.

A combined search for the standard model Higgs boson at s=1.96 TeV

DO Collaboration,Abazov, V.M.,Abbott, B.,Abolins, M.,Acharya, B.S.,Adams, M.,Adams, T.,Aguilo, E.,Ahn, S.H.,Ahsan, M.,Alexeev, G.D.,Alkhazov, G.,Alton, A.,Alverson, G.,Alves, G.A.,Anastasoaie, M.,Ancu North-Holland Pub. Co 2008 Physics letters: B Vol.663 No.1

We present new results of the search for WH->@?νbb@? production in pp@? collisions at a center-of-mass energy of s=1.96 TeV, based on a dataset with integrated luminosity of 0.44 fb<SUP>-1</SUP>. We combine these new results with previously published searches by the D0 collaboration, for WH and ZH production analyzed in the E@?<SUB>T</SUB>bb@? final state, for ZH (->@?<SUP>+</SUP>@?<SUP>-</SUP>bb@?) production, for WH (->WWW) production, and for H (->WW) direct production. No signal-like excess is observed either in the WH analysis or in the combination of all D0 Higgs boson analyses. We set 95% C.L. (expected) upper limits on σ(pp@?->WH)xB(H->bb@?) ranging from 1.6 (2.2) pb to 1.9 (3.3) pb for Higgs boson masses between 105 and 145 GeV, to be compared to the theoretical prediction of 0.13 pb for a Standard Model (SM) Higgs boson with mass m<SUB>H</SUB>=115 GeV. After combination with the other D0 Higgs boson searches, we obtain for m<SUB>H</SUB>=115 GeV an observed (expected) limit 8.5 (12.1) times higher than the SM predicted Higgs boson production cross section. For m<SUB>H</SUB>=160 GeV, the corresponding observed (expected) ratio is 10.2 (9.0).

Flow of a low concentration polyacrylamide fluid solution in a channel with a flat plate obstruction at the entry

Kabir, M.A.,Khan, M.M.K.,Rasul, M.G. The Korean Society of Rheology 2004 Korea-Australia rheology journal Vol.16 No.2

Flow in a channel with an obstruction at the entry can be reverse, stagnant or forward depending on the position of the obstruction. These flow phenomena have potential applications in the control of energy and various flows in process engineering. Parameters that affect this flow inside and around the test channel are the gap (g) between the obstruction geometry and the test channel, the Reynolds number (Re) and the length (L) of the test channel. The influence of these parameters on the flow behavior was investigated using a flat plate obstruction at the entry of the channel. A low concentration polyacrylamide solution (0.018% by weight) showing a powerlaw fluid behavior was used as the fluid in this investigation. The flow phenomena were investigated by the velocity measurement and the flow visualization and their results were compared with numerical simulation. These results of low concentration polyacrylamide solution are also compared with the results of water published elsewhere (Kabir et al., 2003). The maximum reverse flow inside the test channel observed was 20% - 30% of the outside test channel velocity at a g/w (gap to width) ratio of 1 for Reynolds numbers of 1000 to 3500. The influence of the test channel length (L) and the Reynolds number (Re) on the velocity ratio ($V_i$/$V_o$: inside velocity/outside velocity in the test channel) are also presented and discussed here.

Restriction of Metabolizable Energy in Broiler Growers and Its Impact on Grower and Breeder Performance

Sunder, G. Skyam,Kumar, Ch. Vijaya,Panda, A.K.,Raju, M.V.L.N.,Rao, S.V. Rama,Gopinath, N.C.S.,Reddy, M.R. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.8

Metabolizable energy (ME) required for basal metabolism, activity and growth was considered as the criterion for targeting specific increases in body weight (100 g/week) of broiler chicks during the grower phase (5-20 weeks) and its impact was evaluated on breeder performance. Broiler female chicks (460) from a synthetic dam line were randomly distributed to 4 test groups with 23 replicates of 5 birds each and housed in cages. The first group (ME-100) was offered a calculated amount of ME by providing a measured quantity of grower diet (160 g protein and 2,600 kcal ME/kg) which increased with age and weight gain (133-294 kcal/bird/day). The other three groups were offered 10 or 20% less ME (ME-90 and ME-80, respectively) and 10% excess ME (ME-110) over the control group (ME-100). From 21 weeks of age, a single breeder diet (170 g protein and 2,600 kcal ME/kg) was uniformly fed to all groups and the impact of grower ME restriction on breeder performance evaluated up to 58 weeks. The targeted body weight gain of 1,600 g in a 16-week period was achieved by pullets of the ME-100 group almost one week earlier by gaining 8.7 g more weight per week. However, pullets in the ME-90 group gained 1,571 g during the same period, which was closer to the targeted weight. At 20 weeks of age, the conversion efficiency of feed (5.21-5.37), ME (13.9-14.1 kcal/g weight gain) and protein (0.847-0.871 g/g weight gain), eviscerated meat yield, giblet and tibia weights were not influenced by ME restriction, but the weights of abdominal fat and liver were higher with increased ME intake. Reduction of ME by 10% in the grower period significantly delayed sexual maturity (169.3 d), but increased egg production (152.5 /bird) with better persistency. Improved conversion efficiency of feed, ME and protein per g egg content were also observed in this group up to 56 weeks. The fertility and hatchability at 58 weeks of age were higher in the ME-90 group compared to the control and 10% excess ME feeding. In conclusion, the present study revealed the possibility of achieving targeted weight gain in broiler growers by feeding measured quantities of ME during the rearing period with consequential benefits in breeder performance.

Novel anti-apoptotic mechanism of A20 through targeting ASK1 to suppress TNF-induced JNK activation

Won, M,Park, K A,Byun, H S,Sohn, K-C,Kim, Y-R,Jeon, J,Hong, J H,Park, J,Seok, J H,Kim, J M,Yoon, W-H,Jang, I-S,Shen, H M,Liu, Z G,Hur, G M Macmillan Publishers Limited 2010 CELL DEATH AND DIFFERENTIATION Vol.17 No.12

The zinc-finger protein A20 has crucial physiological functions as a dual inhibitor of nuclear factor-κB (NF-κB) activation and apoptosis in tumor necrosis factor (TNF) receptor 1 signaling pathway. Although the molecular basis for the anti-NF-κB function of A20 has been well elucidated, the anti-apoptotic function of A20 is largely unknown. Here, we report a novel mechanism underlying the anti-apoptotic function of A20: A20 blocks TNF-induced apoptosis through suppression of c-jun N-terminal kinase (JNK) by targeting apoptosis signal-regulating kinase1 (ASK1). First, the ectopic expression of A20 drastically inhibits TNF-induced JNK activation and apoptosis in multiple cell types including those deficient of NF-κB activation. Unexpectedly, the blunting effect of A20 on TNF-induced JNK activation is not mediated by affecting the TNFR1 signaling complex formation. Instead, A20 interacts with ASK1, an important MAPKK kinase in the JNK signaling cascade. More importantly, overexpression of wild-type A20, but not of mutant A20 (ZnF4; C624A, C627A), promotes degradation of the ASK1 through the ubiquitin-proteasome system. Taken together, the results from this study reveal a novel anti-apoptotic mechanism of A20 in TNF signaling pathway: A20 binds to ASK1 and mediates ASK1 degradation, leading to suppression of JNK activation and eventually blockage of apoptosis.

Search for Next-to-Minimal Supersymmetric Higgs Bosons in theh→aa→μμμμ,μμττChannels Usingpp¯Collisions ats=1.96 TeV

Abazov, V. M.,Abbott, B.,Abolins, M.,Acharya, B. S.,Adams, M.,Adams, T.,Aguilo, E.,Ahsan, M.,Alexeev, G. D.,Alkhazov, G.,Alton, A.,Alverson, G.,Alves, G. A.,Ancu, L. S.,Andeen, T.,Anzelc, M. S.,Aoki, American Physical Society 2009 Physical Review Letters Vol.103 No.6

<P>We report on a first search for production of the lightest neutral CP-even Higgs boson (h) in the next-to-minimal supersymmetric standard model, where h decays to a pair of neutral pseudoscalar Higgs bosons (a), using 4.2 fb;{-1} of data recorded with the D0 detector at Fermilab. The a bosons are required to either both decay to micro;{+}micro;{-} or one to micro;{+}micro;{-} and the other to tau;{+}tau;{-}. No significant signal is observed, and we set limits on its production as functions of M_{a} and M_{h}.</P>