VEGF gene silencing by cytomegalovirus promoter driven ShRNA expression vector results in vascular development defects in zebrafish

Zebrafish has been generally considered as an excellent model in case of drug screening, disease model establishment, and vertebrate embryonic development study. In this work, the ability of human cytomegalovirus immediate early promoter (CMV promoter)-driven short hairpin RNA (shRNA) expression vector to induce shRNA against VEGF gene in zebrafish was tested, and its effect on vascular development was assed, too. Using RT-qPCR, blood vessel staining, and in situ hybridization, we confirmed certain transcriptional activity and down regulation of gene expression by the vector. In situ hybridization analysis indicated selective inhibition of NRP1 expression in the VEGF gene loss of function model, which might imply in turn that VEGF could not only activate endothelial cells directly but also could contribute to stimulating angiogenesis in vivo by a mechanism that involved up-regulation of its cognate receptor expression in zebrafish. This contributed to a better understanding of molecular mechanisms of cardiovascular development. The system improved the success rate in making inducible knockdown and widened the possibilities for better therapeutic targets in zebrafish.

Angular distribution and conversion of multi-keV L-shell X-ray sources produced from nanosecond laser irradiated thick-foil targets

An experimental study on the angular distribution and conversion of multi-keV X-ray sources produced from 2 ns-duration 527nm laser irradiated thick-foil targets on Shenguang II laser facility (SG-II) is reported. The angular distributions measured in front of the targets can be fitted with the function of f(theta) = alpha+ (1- alpha)cos(beta) theta (theta is the viewing angle relative to the target normal), where alpha = 0.41 +/- 0.014, beta = 0.77 +/- 0.04 for Ti K-shell X-ray Sources (similar to 4.75 keV for Ti K-shell), and alpha = 0.085 +/- 0.06, beta = 0.59 +/- 0.07 for Ag/Pd/Mo L-shell X-ray Sources (2-2.8 keV for Mo L-shell, 2.8-3.5 keV for Pd L-shell, and 3-3.8 keV for Ag L-shell). The isotropy of the angular-distribution of L-shell emission is worse than that of the K-shell emission at larger viewing angle (>70 degrees), due to its larger optical depth (stronger self-absorption) in the cold plasma side lobe Surrounding the central emission region, and in the central hot plasma region (emission region). There is no observable difference in the angular distributions of the L-shell X-ray emission among Ag, Pd, and Mo. The conversion efficiency of Ag/Pd/Mo L-shell X-ray sources is higher than that of the Ti K-shell X-ray sources, but the gain relative to the K-shell emission is not as high as that by using short pulse lasers. The conversion efficiency of the L-shell X-ray sources decrease, with increasing atomic numbers (or X-ray photon energy), similar to the behavior of the K-shell X-ray Source.

Ion acceleration with mixed solid targets interacting with circularly polarized lasers

The interaction of a circularly polarized laser pulse with a mixed solid target containing two species of ions is studied by particle in cell simulations and analytical model. After the interaction tends to be stable, it is demonstrated that the acceleration is more efficient for the heavier ions than that in plasmas containing a single kind of heavy ion and the acceleration efficiency is higher when its proportion is lower. To obtain monoenergetic heavy-ion beams, a sandwich target with a thin mixed ion layer between two light ion layers and a microstructured target are proposed. The influences of parameters of the laser pulse and target on ion acceleration are discussed in detail. It is found that, when the target is thick enough, a cold target is more appropriate for heavy-ion acceleration than a warm target, and the velocity of the reflected heavy ions is proportional to the laser amplitude.

Vacuum heating in the interaction of ultrashort, relativistically strong laser pulses with solid targets

An analytical fluid model for vacuum heating during the oblique incidence by an ultrashort ultraintense p-polarized laser on a solid-density plasma is proposed. The steepening of an originally smooth electron density profile as the electrons are pushed inward by the laser is included self-consistently. It is shown that the electrons being pulled out and then returned to the plasma at the interface layer by the wave field can lead to a phenomenon like wave breaking since the front part of the returning electrons always move slower than the trailing part. This can lead to heating of the plasma at the expense of the wave energy. An estimate for the efficiency of laser energy absorption by the vacuum heating is given. It is also found that for the incident laser intensity parameter, a(L)> 0.5, the absorption rate peaks at an incident angle 45 degrees-52 degrees and it reaches a maximum of 30% at a(L)approximate to 1.5.

Generation of strong quasistatic magnetic fields in interactions of ultraintense and short laser pulses with overdense plasma targets

An analytical fluid model is proposed for the generation of strong quasistatic magnetic fields during normal incidence of a short ultraintense Gaussian laser pulse with a finite spot size on an overdense plasma. The steepening of the electron density profile in the originally homogeneous overdense plasma and the formation of electron cavitation as the electrons are pushed inward by the laser are included self-consistently. It is shown that the appearance of the cavitation plays an important role in the generation of quasistatic magnetic fields: the strong plasma inhomogeneities caused by the formation of the electron cavitation lead to the generation of a strong axial quasistatic magnetic field B-z. In the overdense regime, the generated quasistatic magnetic field increases with increasing laser intensity, while it decreases with increasing plasma density. It is also found that, in a moderately overdense plasma, highly intense laser pulses can generate magnetic fields similar to 100 MG and greater due to the transverse linear mode conversion process.

Blood cells as targets of snake toxins

Snake venoms are mixtures of enzymes and peptides which exert toxicological effects by targeting their substrates or receptors upon envenomation. Snake venom proteins widely affect vascular system including circulating blood cells, coagulation factors, an

RNF220, an E3 ubiquitin ligase that targets Sin3B for ubiquitination

Modification of proteins by ubiquitination plays important roles in various cellular processes. During this process, the target specificity is determined by ubiquitin ligases. Here we identify RNF220 (RING finger protein 220) as a novel ubiquitin ligase for Sin3B. As a conserved RING protein, RNF220 can bind E2 and mediate auto-ubiquitination of itself. Through a yeast two-hybrid screen, we isolated Sin3B as one of its targets, which is a scaffold protein of the Sin3/HDAC (histone deacetylase) corepressor complex. RNF220 specifically interacts with Sin3B both in vitro and in vivo. Sin3B can be regulated by the ubiquitin-proteasome system. Co-expression of RNF220 promotes the ubiquitination and proteasomal degradation of Sin3B. Taken together, these results reveal a new mechanism for regulating the Sin3/HDAC complex. (C) 2010 Elsevier Inc. All rights reserved.

Genome-wide computational identification of microRNAs and their targets in the deep-branching eukaryote Giardia lamblia

Using a combined computational program. we identified 50 potential microRNAs (miRNAs) in Giardia lamblia. one of the most primitive unicellular eukaryotes. These miRNAs are unique to G. lamblia and no homologues have been found in other organisms; miRNAs.

Androgen receptor targets NFKB and TSPI to suppress prostate tumor growth in vivo

The androgen role in the maintenance of prostate epithelium is subject to conflicting opinions. While androgen ablation drives the regression of normal and cancerous prostate, testosterone may cause both proliferation and apoptosis. Several investigators note decreased proliferation and stronger response to chemotherapy of the prostate cancer cells stably expressing androgen receptor (AR), however no mechanistic explanation was offered. In this paper we demonstrate in vivo anti-tumor effect of the AR on prostate cancer growth and identify its molecular mediators. We analyzed the effect of AR on the tumorigenicity of prostate cancer cells. Unexpectedly, the AR-expressing cells formed tumors in male mice at a much lower rate than the AR-negative controls. Moreover, the AR-expressing tumors showed decreased vascularity and massive apoptosis. AR expression lowered the angiogenic potential of cancer cells, by increasing secretion of an anti-angiogenic protein, thrombospondin-1. AR activation caused a decrease in RelA, a subunit of the pro-survival transcription factor NF kappa B, reduced its nuclear localization and transcriptional activity. This, in turn, diminished the expression of its anti-apoptotic targets, Bcl-2 and IL-6. Increased apoptosis within AR-expressing tumors was likely due to the NF kappa B suppression, since it was restricted to the cells lacking nuclear (active) NF kappa B. Thus we for the first time identified combined decrease of NF kappa B and increased TSP1 as molecular events underlying the AR anti-tumor activity in vivo. Our data indicate that intermittent androgen ablation is preferable to continuous withdrawal, a standard treatment for early-stage prostate cancer. (C) 2007 Wiley-Liss, Inc.