Multi-Color Coronagraph Experiment in a Vacuum Testbed with a Binary Shaped Pupil Mask

We conducted a number of multi-band coronagraph experiments using a vacuum chamber and a binary-shaped pupil mask which in principle should work at all wavelengths, in the context of the research and development on a coronagraph to observe extra-solar planets (exoplanets) directly. The aim of this work is to demonstrate that subtraction of Point Spread Function (PSF) and multi-band experiments using a binary-shaped pupil mask coronagraph would help improve the contrast in the observation of exoplanets. A checkerboard mask, a kind of binary-shaped pupil mask, was used. We improved the temperature stability by installing the coronagraph optics in a vacuum chamber, controlling the temperature of the optical bench, and covering the vacuum chamber with thermal insulation layers. We evaluated how much the PSF subtraction contributes to the high contrast observation by subtracting the images obtained through the coronagraph. We also carried out multi- band experiments in order to demonstrate a more realistic observation using Super luminescent Light Emitting Diodes (SLEDs) with center wavelengths of 650nm, 750nm, 800nm and 850nm. A contrast of 2.3x10-7 was obtained for the raw coronagraphic image and a contrast of 1.3x10-9 was achieved after PSF subtraction with a He-Ne laser at 632.8nm wavelength. Thus, the contrast was improved by around two orders of magnitude from the raw contrast by subtracting the PSF. We achieved contrasts of 3.1x10-7, 1.1x10-6, 1.6x10-6 and 2.5x10-6 at the bands of 650nm, 750nm, 800nm and 850nm, respectively, in the multi-band experiments. The results show that contrast within each of the wavelength bands was significantly improved compared with non-coronagraphic optics. We demonstrated PSF subtraction is potentially beneficial for improving contrast of the coronagraph, and this coronagraph produces a significant improvement in contrast with multi-band light sources.Comment: 14 pages, 7 figures, accepted for publication in PAS

An Endogenous Murine Leukemia Viral Genome Contaminant in a Commercial RT-PCR Kit is Amplified Using Standard Primers for XMRV

During pilot studies to investigate the presence of viral RNA of xenotropic murine leukemia virus (MLV)-related virus (XMRV) infection in sera from chronic fatigue syndrome (CFS) patients in Japan, a positive band was frequently detected at the expected product size in negative control samples when detecting a partial gag region of XMRV using a one-step RT-PCR kit. We suspected that the kit itself might have been contaminated with small traces of endogenous MLV genome or XMRV and attempted to evaluate the quality of the kit in two independent laboratories. We purchased four one-step RT-PCR kits from Invitrogen, TaKaRa, Promega and QIAGEN in Japan. To amplify the partial gag gene of XMRV or other MLV-related viruses, primer sets (419F and 1154R, and GAG-I-F and GAG-I-R) which have been widely used in XMRV studies were employed. The nucleotide sequences of the amplicons were determined and compared with deposited sequences of a polytropic endogenous MLV (PmERV), XMRV and endogenous MLV-related viruses derived from CFS patients. We found that the enzyme mixtures of the one-step RT-PCR kit from Invitrogen were contaminated with RNA derived from PmERV. The nucleotide sequence of a partial gag region of the contaminant amplified by RT-PCR was nearly identical (99.4% identity) to a PmERV on chromosome 7 and highly similar (96.9 to 97.6%) to recently identified MLV-like viruses derived from CFS patients. We also determined the nucleotide sequence of a partial env region of the contaminant and found that it was almost identical (99.6%) to the PmERV. In the investigation of XMRV infection in patients of CFS and prostate cancer, researchers should prudently evaluate the test kits for the presence of endogenous MLV as well as XMRV genomes prior to PCR and RT-PCR tests

Integrity testing of Planova™ BioEX virus removal filters used in the manufacture of biological products

AbstractConfirmation of virus filter integrity is crucial for ensuring the safety of biological products. Two main types of virus filter defects may produce inconsistent and undesirable performance in virus removal: improper pore-size distribution across the membrane; and specific damage, such as tears, broken fibers, or pinholes. Two integrity tests are performed on each individual filter manufactured by Asahi Kasei Medical to ensure the absence of these defects prior to shipment. In this study, we verified that typical usage of Planova™ BioEX filters would not improperly shift the pore-size distribution. Damage occurring during shipment and use (e.g., broken fibers or pinholes) can be detected by end-users with sufficient sensitivity using air–water diffusion based leakage tests. We prepared and tested filters with model pinhole defects of various sizes to develop standard acceptance criteria for the leakage test relative to porcine parvovirus infectivity logarithmic reduction values (LRVs). Our results demonstrate that pinhole defects at or below a certain size for each effective filter surface area have no significant impact on the virus LRV. In conclusion the leakage test is sufficiently sensitive to serve as the sole end-user integrity test for Planova™ BioEX filters, facilitating their use in biopharmaceuticals manufacturing