Species D human adenovirus type 9 exhibits better virus-spread ability for antitumor efficacy among alternative serotypes

Species C human adenovirus serotype 5 (HAdV-C5) is widely used as a vector for cancer gene therapy, because it efficiently transduces target cells. A variety of HAdV-C5 vectors have been developed and tested in vitro and in vivo for cancer gene therapy. While clinical trials with HAdV-C5 vectors resulted in effective responses in many cancer patients, administration of HAdV-C5 vectors to solid tumors showed responses in a limited area. A biological barrier in tumor mass is considered to hinder viral spread of HAdV-C5 vectors from infected cells. Therefore, efficient virus-spread from an infected tumor cell to surrounding tumor cells is required for successful cancer gene therapy. In this study, we compared HAdV-C5 to sixteen other HAdV serotypes selected from species A to G for virus-spread ability in vitro. HAdV-D9 showed better virus-spread ability than other serotypes, and its viral progeny were efficiently released from infected cells during viral replication. Although the HAdV-D9 fiber protein contains a binding site for coxsackie B virus and adenovirus receptor (CAR), HAdV-D9 showed expanded tropism for infection due to human CAR (hCAR)-independent attachment to target cells. HAdV-D9 infection effectively killed hCAR-negative cancer cells as well as hCAR-positive cancer cells. These results suggest that HADV-D9, with its better virus-spread ability, could have improved therapeutic efficacy in solid tumors compared to HAdV-C5

A multi targeting conditionally replicating adenovirus displays enhanced oncolysis while maintaining expression of immunotherapeutic agents

Studies have demonstrated that oncolytic adenoviruses based on a 24 base pair deletion in the viral E1A gene (D24) may be promising therapeutics for treating a number of cancer types. In order to increase the therapeutic potential of these oncolytic viruses, a novel conditionally replicating adenovirus targeting multiple receptors upregulated on tumors was generated by incorporating an Ad5/3 fiber with a carboxyl terminus RGD ligand. The virus displayed full cytopathic effect in all tumor lines assayed at low titers with improved cytotoxicity over Ad5-RGD D24, Ad5/3 D24 and an HSV oncolytic virus. The virus was then engineered to deliver immunotherapeutic agents such as GM-CSF while maintaining enhanced heterogenic oncolysis

X-ray Properties of the Weak Seyfert 1 Nucleus in NGC 4639

We obtained observations of NGC 4639 with ASCA in order to investigate its mildly active Seyfert 1 nucleus at hard X-ray energies. Koratkar et al. (1995) have previously shown that the nucleus is a pointlike source in the ROSAT soft X-ray band. We detected in the 2-10 keV band a compact central source with a luminosity of 8.3E+40 erg/s. Comparison of the ASCA data with archival data taken with the Einstein and ROSAT satellites shows that the nucleus varies on timescales of months to years. The variability could be intrinsic, or it could be caused by variable absorption. More rapid variability, on a timescale of \~10^4 s, may be present in the ASCA data. The spectrum from 0.5 to 10 keV is well described by a model consisting of a lightly absorbed (N_H = 7.3E+20 cm^-2) power law with a photon index of 1.68. We find no evidence for significant emission from a thermal plasma; if present, it can account for no more than 25% of the flux in the 0.5-2.0 keV band. The limited photon statistics of our data do not allow us to place significant limits on the presence of iron K emission. (abridged)Comment: To appear in The Astrophysical Journal. LaTex, 18 pages including embedded figures and table

Detection of an iron K Emission Line from the LINER NGC 4579

We present the results of an ASCA observation of the LINER NGC 4579. A point-like X-ray source is detected at the nucleus with a 2-10 keV luminosity of 1.5x10^41 ergs/s assuming a distance of 16.8 Mpc. The X-ray spectrum is represented by a combination of a power-law with a photon index of ~1.7 and soft thermal component with kT~0.9 keV. An iron K emission line is detected at 6.73+/-0.13 keV (rest frame) with an equivalent width of 490 +180/-190 eV and is statistically significant at more than 99.9 % confidence. The line center energy is consistent with Helium-like iron and is significantly higher than 6.4 keV which is expected from fluorescence by "cold" (or a lower ionization state of) iron. The iron line profile shows no significant red tail in contrast to Seyfert 1 galaxies although the statistics are limited. The line center energy, equivalent width, and profile are consistent with an origin in an ionized accretion disk. However the large mass accretion rate necessary to ionize the accretion disk is not consistent with the observed luminosity and normal accretion models.Comment: 15 pages, 5 figures, to appear in The Astrophysical Journa

Evaluation of adenovirus capsid labeling versus transgene expression

Adenoviral vectors have been utilized for a variety of gene therapy applications. Our group has incorporated bioluminescent, fluorographic reporters, and/or suicide genes within the adenovirus genome for analytical and/or therapeutic purposes. These molecules have also been incorporated as capsid components. Recognizing that incorporations at either locale yield potential advantages and disadvantages, our report evaluates the benefits of transgene incorporation versus capsid incorporation. To this end, we have genetically incorporated firefly luciferase within the early region 3 or at minor capsid protein IX and compared vector functionality by means of reporter readout

Lung of Fgf10-CRISPR mosaic mouse

CRISPR/Cas9-mediated gene editing often generates founder generation (F0) mice that exhibit somatic mosaicism in the targeted gene(s). It has been known that Fibroblast growth factor 10 (Fgf10)-null mice exhibit limbless and lungless phenotypes, while intermediate limb phenotypes (variable defective limbs) are observed in the Fgf10-CRISPR F0 mice. However, how the lung phenotype in the Fgf10-mosaic mutants is related to the limb phenotype and genotype has not been investigated. In this study, we examined variable lung phenotypes in the Fgf10-targeted F0 mice to determine if the lung phenotype was correlated with percentage of functional Fgf10 genotypes. Firstly, according to a previous report, Fgf10-CRISPR F0 embryos on embryonic day 16.5 (E16.5) were classified into three types: type I, no limb; type II, limb defect; and type III, normal limbs. Cartilage and bone staining showed that limb truncations were observed in the girdle, (type I), stylopodial, or zeugopodial region (type II). Deep sequencing of the Fgf10-mutant genomes revealed that the mean proportion of codons that encode putative functional FGF10 was 8.3 ± 6.2% in type I, 25.3 ± 2.7% in type II, and 54.3 ± 9.5% in type III (mean ± standard error of the mean) mutants at E16.5. Histological studies showed that almost all lung lobes were absent in type I embryos. The accessory lung lobe was often absent in type II embryos with other lobes dysplastic. All lung lobes formed in type III embryos. The number of terminal tubules was significantly lower in type I and II embryos, but unchanged in type III embryos. To identify alveolar type 2 epithelial (AECII) cells, known to be reduced in the Fgf10-heterozygous mutant, immunostaining using anti-surfactant protein C (SPC) antibody was performed: In the E18.5 lungs, the number of AECII was correlated to the percentage of functional Fgf10 genotypes. These data suggest the Fgf10 gene dose-related loss of the accessory lobe and decrease in the number of alveolar type 2 epithelial cells in mouse lung. Since dysfunction of AECII cells has been implicated in the pathogenesis of parenchymal lung diseases, the Fgf10-CRISPR F0 mouse would present an ideal experimental system to explore it

ASCA Observations of "Type 2" LINERs: Evidence for a Stellar Source of Ionization

We present ASCA observations of LINERs without broad H$\alpha$ emission in their optical spectra. The sample of "type 2" LINERs consists of NGC 404, 4111, 4192, 4457, and 4569. We have detected X-ray emission from all the objects except for NGC 404; among the detected objects are two so-called transition objects (NGC 4192 and NGC 4569), which have been postulated to be composite nuclei having both an HII region and a LINER component. The images of NGC 4111 and NGC 4569 in the soft (0.5-2 keV) and hard (2-7 keV) X-ray bands are extended on scales of several kpc. The X-ray spectra of NGC 4111, NGC 4457 and NGC 4569 are well fitted by a two-component model that consists of soft thermal emission with $kT\sim0.65$ keV and a hard component represented by a power law (photon index $\sim$ 2) or by thermal bremsstrahlung emission ($kT\sim$ several keV). The extended hard X-rays probably come from discrete sources, while the soft emission most likely originates from hot gas produced by active star formation in the host galaxy. We have found no clear evidence for the presence of active galactic nuclei (AGNs) in the sample. If an AGN component is the primary ionization source of the optical emission lines, then it must be heavily obscured with a column density significantly larger than $10^{23}$ cm$^{-2}$. Alternatively, the optical emission could be ionized by a population of exceptionally hot stars.Comment: 13 pages, 5 figures, emulateapj.sty, Accepted for publication in the Astrophysical Journa

Fgf10-CRISPR mosaic mutants demonstrate the gene dose-related loss of the accessory lobe and decrease in the number of alveolar type 2 epithelial cells in mouse lung

CRISPR/Cas9-mediated gene editing often generates founder generation (F0) mice that exhibit somatic mosaicism in the targeted gene(s). It has been known thatFibroblast growth factor 10(Fgf10)-null mice exhibit limbless and lungless phenotypes, while intermediate limb phenotypes (variable defective limbs) are observed in theFgf10-CRISPR F0 mice. However, how the lung phenotype in theFgf10-mosaic mutants is related to the limb phenotype and genotype has not been investigated. In this study, we examined variable lung phenotypes in theFgf10-targeted F0 mice to determine if the lung phenotype was correlated with percentage of functionalFgf10genotypes. Firstly, according to a previous report,Fgf10-CRISPR F0 embryos on embryonic day 16.5 (E16.5) were classified into three types: type I, no limb; type II, limb defect; and type III, normal limbs. Cartilage and bone staining showed that limb truncations were observed in the girdle, (type I), stylopodial, or zeugopodial region (type II). Deep sequencing of theFgf10-mutant genomes revealed that the mean proportion of codons that encode putative functional FGF10 was 8.3 +/- 6.2% in type I, 25.3 +/- 2.7% in type II, and 54.3 +/- 9.5% in type III (mean +/- standard error of the mean) mutants at E16.5. Histological studies showed that almost all lung lobes were absent in type I embryos. The accessory lung lobe was often absent in type II embryos with other lobes dysplastic. All lung lobes formed in type III embryos. The number of terminal tubules was significantly lower in type I and II embryos, but unchanged in type III embryos. To identify alveolar type 2 epithelial (AECII) cells, known to be reduced in theFgf10-heterozygous mutant, immunostaining using anti-surfactant protein C (SPC) antibody was performed: In the E18.5 lungs, the number of AECII was correlated to the percentage of functionalFgf10genotypes. These data suggest theFgf10gene dose-related loss of the accessory lobe and decrease in the number of alveolar type 2 epithelial cells in mouse lung. Since dysfunction of AECII cells has been implicated in the pathogenesis of parenchymal lung diseases, theFgf10-CRISPR F0 mouse would present an ideal experimental system to explore it

The Type Ic Hypernova SN 2002ap

Photometric and spectroscopic data of the energetic Type Ic supernova (SN) 2002ap are presented, and the properties of the SN are investigated through models of its spectral evolution and its light curve. The SN is spectroscopically similar to the "hypernova" SN 1997ef. However, its kinetic energy [$\sim (4-10) \times 10^{51}$ erg] and the mass ejected (2.5-5 $M_{\odot}$) are smaller, resulting in a faster-evolving light curve. The SN synthesized $\sim 0.07 M_{\odot}$ of $^{56}$Ni, and its peak luminosity was similar to that of normal SNe. Brightness alone should not be used to define a hypernova, whose defining character, namely very broad spectral features, is the result of a high kinetic energy. The likely main-sequence mass of the progenitor star was 20-25 $M_{\odot}$, which is also lower than that of both hypernovae SNe 1997ef and 1998bw. SN 2002ap appears to lie at the low-energy and low-mass end of the hypernova sequence as it is known so far. Observations of the nebular spectrum, which is expected to dominate by summer 2002, are necessary to confirm these values.Comment: 10 pages, 4 figures, accepted for publication in ApJL, 30 April 2002 (minor changes to match the accepted version, with figures being colored