The Carboxyl-Terminal Amino Acids Render Pro-Human LC3B Migration Similar to Lipidated LC3B in SDS-PAGE

LC3 is widely used marker for macroautophagy assays. After translation pro-LC3 is processed by Atg4 to expose C-terminal glycine residue for downstream conjugation reactions to accomplish the conversion of LC3-I to LC3-II. SDS-PAGE based Western blot (Wb) is generally utilized to quantify LC3-II levels where the LC3-I band migrates slower than LC3-II. We found that pro-human LC3B migrated at similar rate as LC3B-II in SDS-PAGE. The carboxyl-terminal five amino acids, particularly Lysine122 and Leucine123 of human LC3B play a major role in the faster migration of unprocessed LC3B, rendering it indistinguishable from LC3B-II in Wb assays. The unique faster migration of unprocessed LC3B than LC3B-I is also revealed in mouse LC3B, rat LC3B and rat LC3 but not in human LC3C. Our findings for the first time define pro-LC3 migration patterns for LC3 family member from human, mouse and rat species in SDS-PAGE. These findings provide a reference for pro-LC3 band patterns when Atg4 function is inhibited. © 2013 Wang et al

A population-based case-control study on social factors and risk of testicular germ cell tumours

Objectives Incidence rates for testicular cancer have risen over the last few decades. Findings of an association between the risk of testicular cancer and social factors are controversial. The association of testicular cancer and different indicators of social factors were examined in this study.<p></p> Design Case–control study.<p></p> Setting Population-based multicentre study in four German regions (city states Bremen and Hamburg, the Saarland region and the city of Essen).<p></p> Participants The study included 797 control participants and 266 participants newly diagnosed with testicular cancer of which 167 cases were classified as seminoma and 99 as non-seminoma. The age of study participants ranged from 15 to 69 years.<p></p> Methods Social position was classified by educational attainment level, posteducational training, occupational sectors according to Erikson-Goldthorpe-Portocarrero (EGP) and the socioeconomic status (SES) on the basis of the International SocioEconomic Index of occupational status (ISEI). ORs and corresponding 95% CIs (95% CIs) were calculated for the whole study sample and for seminoma and non-seminoma separately.<p></p> Results Testicular cancer risk was modestly increased among participants with an apprenticeship (OR=1.7 (95% CI 1.0 to 2.8)) or a university degree (OR=1.6 (95% CI 0.9 to 2.8)) relative to those whose education was limited to school. Analysis of occupational sectors revealed an excess risk for farmers and farm-related occupations. No clear trend was observed for the analyses according to the ISEI-scale.<p></p> Conclusions Social factors based on occupational measures were not a risk factor for testicular cancer in this study. The elevated risk in farmers and farm-related occupations warrants further research including analysis of occupational exposures.<p></p&gt

The Gamma-Ray Imaging Spectrometer (GRIS): A new balloon-borne experiment for gamma-ray line astronomy

High resolution gamma-ray spectroscopy is a relatively new field that holds great promise for further understanding of high energy astrophysical processes. When the high resolution gamma-ray spectrometer (GRSE) was removed from the GRO payload, a balloon program was initiated to permit continued development and improvement of instrumentation in this field, as well as continued scientific observations. The Gamma-Ray Imaging Spectrometer (GRIS) is one of the experiments selected as part of this program. The instrument contains a number of new and innovative features that are expected to produce a significant improvement in source location accuracy and sensitivity over previous balloon and satellite experiments

A coded aperture imaging system optimized for hard X-ray and gamma ray astronomy

A coded aperture imaging system was designed for the Gamma-Ray imaging spectrometer (GRIS). The system is optimized for imaging 511 keV positron-annihilation photons. For a galactic center 511-keV source strength of 0.001 sq/s, the source location accuracy is expected to be + or - 0.2 deg

E1-Like Activating Enzyme Atg7 Is Preferentially Sequestered into p62 Aggregates via Its Interaction with LC3-I

p62 is constitutively degraded by autophagy via its interaction with LC3. However, the interaction of p62 with LC3 species in the context of the LC3 lipidation process is not specified. Further, the p62-mediated protein aggregation's effect on autophagy is unclear. We systemically analyzed the interactions of p62 with all known Atg proteins involved in LC3 lipidation. We find that p62 does not interact with LC3 at the stages when it is being processed by Atg4B or when it is complexed or conjugated with Atg3. p62 does interact with LC3-I and LC3-I:Atg7 complex and is preferentially recruited by LC3-II species under autophagic stimulation. Given that Atg4B, Atg3 and LC3-Atg3 are indispensable for LC3-II conversion, our study reveals a protective mechanism for Atg4B, Atg3 and LC3-Atg3 conjugate from being inappropriately sequestered into p62 aggregates. Our findings imply that p62 could potentially impair autophagy by negatively affecting LC3 lipidation and contribute to the development of protein aggregate diseases. © 2013 Gao et al

Bada Bang, Bada Boom: Dispersal of fall migrating cormorants to protect sportfish on Oneida Lake, New York

The interior population of double-crested cormorants (Phalacrocorax auritus) continues to increase. As a result, conflicts between human interests and cormorants have intensified. The impacts of nesting, roosting, and migrating cormorants include predation at aquaculture facilities, interspecific competition with rare species including common terns {Sterna hirundo), and impacts to private property. In addition, heightened public and scientific debate regarding cormorant impacts on sportfish has accelerated the need for effective, socially acceptable methods for managing local conflicts. In 1998 and 1999, USDA, APHIS, Wildlife Services; the New York State Department of Environmental Conservation; and USGS, New York Fish and Wildlife Research Cooperative collaborated on a pilot project to investigate prospective methods and strategies for reducing predation of yellow perch (Perca flavescens) and walleye (Stizostedion vitreum) by fall migrating cormorants on Oneida Lake, New York. The goal of the cooperative program was to reduce stopover time, thereby reducing predation on fish stocks by migrating cormorants. A lakewide integrated non-lethal harassment and roost management program was initiated to disperse local and migrating cormorants from September to October each year. A variety of methods were used to manage cormorants including: electronic guards, propane cannons, mylar tape, human effigies, pyrotechnics, and dispersing birds with a boat. Harassment was focused on birds loafing on the water as well as day and night roosting sites (8 day and 2 night roosts). A total of 52,840 cormorants was dispersed (in many cases individual birds were harassed multiple times) using 1,518 pyrotechnics and 649 staff hours on the lake during the 2 years of the program. Surveys documented a 61%-98% reduction of the cormorants population on Oneida Lake compared to mean counts during the same time period from 1995-1997. Estimated total annual fish consumption by cormorants was reduced by 30% in 1998. Some off-site impacts of the program were seen with increased flocks of cormorants documented on nearby lakes

Unintended spread of a biosafety level 2 recombinant retrovirus

<p>Abstract</p> <p>Background</p> <p>Contamination of vertebrate cell lines with animal retroviruses has been documented repeatedly before. Although such viral contaminants can be easily identified with high sensitivity by PCR, it is impossible to screen for all potential contaminants. Therefore, we explored two novel methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant.</p> <p>Results</p> <p>The first hint for the presence of contaminating retroviruses in one of our cell lines was obtained by electron microscopy of exosome-like vesicles released from the supernatants of transfected 293T cells. Random amplification of particle associated RNAs (PAN-PCR) from supernatant of contaminated 293T cells and sequencing of the amplicons revealed several nucleotide sequences showing highest similarity to either murine leukemia virus (MuLV) or squirrel monkey retrovirus (SMRV). Subsequent mass spectrometry analysis confirmed our findings, since we could identify several peptide sequences originating from monkey and murine retroviral proteins. Quantitative PCRs were established for both viruses to test currently cultured cell lines as well as liquid nitrogen frozen cell stocks. Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.</p> <p>Conclusion</p> <p>The high degree of nucleotide identity suggests unintended spread of a biosafety level 2 recombinant virus, which could also affect the risk assessment of gene-modified organisms released from contaminated cell cultures. The study further indicates that both mass spectrometry and PAN-PCR are powerful methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant. Both methods might be useful tools for testing cell lines before using them for critical purposes.</p

Biomarkers of the L-arginine / dimethylarginine / nitric oxide pathway in people with chronic airflow obstruction and obstructive sleep apnoea

Background: Chronic obstructive pulmonary disease (COPD) and obstructive sleep apnoea (OSA) are common chronic diseases that are associated with chronic and intermittent hypoxemia, respectively. Patients affected by the overlap of COPD and OSA have a particularly unfavourable prognosis. The L-arginine/nitric oxide (NO) pathway plays an important role in regulating pulmonary vascular function. Asymmetric (ADMA) and symmetric dimethylarginine (SDMA) interfere with NO production. Methods: We analysed the serum concentrations of ADMA, SDMA, L-arginine, L-citrulline, and L-ornithine in a large sample of the Icelandic general population together with chronic airflow obstruction (CAO), a key physiological marker of COPD that was assessed by post-bronchodilator spirometry (FEV1/FVC 0.5. SDMA was significantly higher in individuals with CAO (0.518 [0.461–0.616] vs. 0.494 [0.441–0.565] µmol/L; p = 0.005), but ADMA was not. However, ADMA was significantly associated with decreasing FEV1 percent predicted among those with CAO (p = 0.002). ADMA was 0.50 (0.44–0.56) µmol/L in MAP ≤ 0.5 versus 0.52 (0.46–0.58) µmol/L in MAP > 0.5 (p = 0.008). SDMA was 0.49 (0.44–0.56) µmol/L versus 0.51 (0.46–0.60) µmol/L, respectively (p = 0.004). The highest values for ADMA and SDMA were observed in individuals with overlap of CAO and MAP > 0.5, which was accompanied by lower L-citrulline levels. Conclusions: The plasma concentrations of ADMA and SDMA are elevated in COPD patients with concomitant intermittent hypoxaemia. This may account for impaired pulmonary NO production, enhanced pulmonary vasoconstriction, and disease progression