Rapid Detection and Quantification of Triacylglycerol by HPLC–ELSD in \u3ci\u3eChlamydomonas reinhardtii\u3c/i\u3e and \u3ci\u3eChlorella\u3c/i\u3e Strains

Triacylglycerol (TAG) analysis and quantification are commonly performed by first obtaining a purified TAG fraction from a total neutral lipid extract using thinlayer chromatography (TLC), and then analyzing the fatty acid composition of the purified TAG fraction by gas chromatography (GC). This process is time-consuming, labor intensive and is not suitable for analysis of small sample sizes or large numbers. A rapid and efficient method for monitoring oil accumulation in algae using high performance liquid chromatography for separation of all lipid classes combined with detection by evaporative light scattering (HPLC–ELSD) was developed and compared to the conventional TLC/GC method. TAG accumulation in two Chlamydomonas reinhardtii (21 gr and CC503) and three Chlorella strains (UTEX 1230, CS01 and UTEX 2229) grown under conditions of nitrogen depletion was measured. The TAG levels were found to be 3–6 % DW (Chlamydomonas strains) and 7–12 % DW (Chlorella strains) respectively by both HPLC–ELSD and TLC/GC methods. HPLC–ELSD resolved the major lipid classes such as carotenoids, TAG, diacylglycerol (DAG), free fatty acids, phospholipids, and galactolipids in a 15-min run. Quantitation of TAG content was based on comparison to calibration curves of trihexadecanoin (16:0 TAG) and trioctadecadienoin (18:2 TAG) and showed linearity from 0.2 to 10 lg. Algal TAG levels \u3e0.5 lg/g DW were detectable by this method. Furthermore TAG content in Chlorella kessleri UTEX 2229 could be detected. TAG as well as DAG and TAG content were estimated at 1.6 % DWby HPLC–ELSD, while it was undetectable by TLC/GC method

Statin use and risk of community acquired pneumonia in older people: population based case-control study

Objective To test the hypothesis that hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) may decrease the risk of community acquired pneumonia

SMASHing the LMC: A Tidally-induced Warp in the Outer LMC and a Large-scale Reddening Map

We present a study of the three-dimensional (3D) structure of the Large Magellanic Cloud (LMC) using ~2.2 million red clump (RC) stars selected from the Survey of the MAgellanic Stellar History. To correct for line-of-sight dust extinction, the intrinsic RC color and magnitude and their radial dependence are carefully measured by using internal nearly dust-free regions. These are then used to construct an accurate 2D reddening map (165 square degrees with ~10 arcmin resolution) of the LMC disk and the 3D spatial distribution of RC stars. An inclined disk model is fit to the 2D distance map yielding a best-fit inclination angle i = 25.86(+0.73,-1.39) degrees with random errors of +\-0.19 degrees and line-of-nodes position angle theta = 149.23(+6.43,-8.35) degrees with random errors of +/-0.49 degrees. These angles vary with galactic radius, indicating that the LMC disk is warped and twisted likely due to the repeated tidal interactions with the Small Magellanic Cloud (SMC). For the first time, our data reveal a significant warp in the southwestern part of the outer disk starting at rho ~ 7 degrees that departs from the defined LMC plane up to ~4 kpc toward the SMC, suggesting that it originated from a strong interaction with the SMC. In addition, the inner disk encompassing the off-centered bar appears to be tilted up to 5-15 degrees relative to the rest of the LMC disk. These findings on the outer warp and the tilted bar are consistent with the predictions from the Besla et al. simulation of a recent direct collision with the SMC.Comment: 25 pages, 15 figures, published in Ap

Near-atomic, non-icosahedrally averaged structure of giant virus Paramecium bursaria chlorella virus 1

Giant viruses are a large group of viruses that infect many eukaryotes. Although components that do not obey the overall icosahedral symmetry of their capsids have been observed and found to play critical roles in the viral life cycles, identities and high-resolution structures of these components remain unknown. Here, by determining a near-atomic-resolution, five-fold averaged structure of Parameciumbursaria chlorella virus 1, we unexpectedly found the viral capsid possesses up to five major capsid protein variants and a penton protein variant. These variants create varied capsidmicroenvironments for the associations of fibers, a vesicle, and previously unresolved minor capsid proteins. Our structure reveals the identities and atomic models of the capsid components that do not obey the overall icosahedral symmetry and leads to a model for how these components are assembled and initiate capsid assembly, and this model might be applicable to many other giant viruses

HST/ACS Emission Line Imaging of Low Redshift 3CR Radio Galaxies I: The Data

We present 19 nearby (z<0.3) 3CR radio galaxies imaged at low- and high-excitation as part of a Cycle 15 Hubble Space Telescope snapshot survey with the Advanced Camera for Surveys. These images consist of exposures of the H-alpha (6563 \AA, plus [NII] contamination) and [OIII] 5007 \AA emission lines using narrow-band linear ramp filters adjusted according to the redshift of the target. To facilitate continuum subtraction, a single-pointing 60 s line-free exposure was taken with a medium-band filter appropriate for the target's redshift. We discuss the steps taken to reduce these images independently of the automated recalibration pipeline so as to use more recent ACS flat-field data as well as to better reject cosmic rays. We describe the method used to produce continuum-free (pure line-emission) images, and present these images along with qualitative descriptions of the narrow-line region morphologies we observe. We present H-alpha+[NII] and [OIII] line fluxes from aperture photometry, finding the values to fall expectedly on the redshift-luminosity trend from a past HST/WFPC2 emission line study of a larger, generally higher redshift subset of the 3CR. We also find expected trends between emission line luminosity and total radio power, as well as a positive correlation between the size of the emission line region and redshift. We discuss the associated interpretation of these results, and conclude with a summary of future work enabled by this dataset.Comment: 18 pages, 12 figures, accepted for publication in ApJ

The N-glycan structures of the antigenic variants of chlorovirus PBCV-1 major capsid protein help to identify the virus-encoded glycosyltransferases

The chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) is a large dsDNA virus that infects the microalga Chlorella variabilis NC64A. Unlike most other viruses, PBCV-1 encodes most, if not all, of the machinery required to glycosylate its major capsid protein (MCP). The structures of the four N-linked glycans from the PBCV-1 MCP consist of nonasaccharides, and similar glycans are not found elsewhere in the three domains of life. Here, we identified the roles of three virus-encoded glycosyltransferases (GTs) that have four distinct GT activities in glycan synthesis. Two of the three GTs were previously annotated as GTs but the third GT was identified in this study. We determined the GT functions by comparing the wild-type glycan structures from PBCV-1 with those from a set of PBCV-1 spontaneous GT genes mutants resulting in antigenic variants having truncated glycan structures. According to our working model, the virus gene a064r encodes a GT with three domains: domain 1 has a β-L-rhamnosyltransferase activity, domain 2 has an α -L-rhamnosyltransferase activity and domain 3 is a methyltransferase that decorates two positions in the terminal α -L-rhamnose (Rha) unit. The a075l gene encodes a β -xylosyltransferase that attaches the distal D-xylose (Xyl) unit to the L-fucose (Fuc) that is part of the conserved N-glycan core region. Lastly, gene a071r encodes a GT that is involved in the attachment of a semiconserved element, α-D-Rha, to the same L-Fuc in the core region. Our results uncover GT activities that assemble four of the nine residues of the PBCV-1 MCP N-glycans. Includes supplemental material