Intranasal immunization with inactivated influenza virus enhances immune responses to coadministered simian-human immunodeficiency virus-like particle antigens

Intranasal immunization with inactivated influenza virus vaccine can provide protective immunity, whereas many other antigens are less effective when used for mucosal immunization. To determine whether influenza virus could enhance immune responses to an antigen coadministered to a mucosal surface, we studied the intranasal immunization of mice with a mixture of simian-human immunodeficiency virus (SHIV) virus-like particles (VLPs) and inactivated influenza virus. Compared to mice immunized with SHIV VLPs alone, mice coimmunized with SHIV VLPs and inactivated influenza virus showed significant increases in serum immunoglobulin G (IgG) and mucosal IgA antibodies specific to the human immunodeficiency virus envelope protein, neutralizing activities, numbers of gamma interferon- and interleukin 4-secreting lymphocytes, and cytotoxic-T-lymphocyte activities. The levels of enhancement of immune response by coimmunization with inactivated influenza virus were equivalent to those induced by inclusion of immunostimulatory CpG oligodeoxynucleotides (CpG DNA). We also observed that SHIV VLPs bind to influenza virus virions, forming mixed aggregates. These results indicate that inactivated influenza virus can play a role as a mucosal adjuvant to coadministered antigens.     

iLIR: A web resource for prediction of Atg8-family interacting proteins

Macroautophagy was initially considered to be a nonselective process for bulk breakdown of cytosolic material. However, recent evidence points toward a selective mode of autophagy mediated by the so-called selective autophagy receptors (SARs). SARs act by recognizing and sorting diverse cargo substrates (e.g., proteins, organelles, pathogens) to the autophagic machinery. Known SARs are characterized by a short linear sequence motif (LIR-, LRS-, or AIM-motif) responsible for the interaction between SARs and proteins of the Atg8 family. Interestingly, many LIR-containing proteins (LIRCPs) are also involved in autophagosome formation and maturation and a few of them in regulating signaling pathways. Despite recent research efforts to experimentally identify LIRCPs, only a few dozen of this class of—often unrelated—proteins have been characterized so far using tedious cell biological, biochemical, and crystallographic approaches. The availability of an ever-increasing number of complete eukaryotic genomes provides a grand challenge for characterizing novel LIRCPs throughout the eukaryotes. Along these lines, we developed iLIR, a freely available web resource, which provides in silico tools for assisting the identification of novel LIRCPs. Given an amino acid sequence as input, iLIR searches for instances of short sequences compliant with a refined sensitive regular expression pattern of the extended LIR motif (xLIR-motif) and retrieves characterized protein domains from the SMART database for the query. Additionally, iLIR scores xLIRs against a custom position-specific scoring matrix (PSSM) and identifies potentially disordered subsequences with protein interaction potential overlapping with detected xLIR-motifs. Here we demonstrate that proteins satisfying these criteria make good LIRCP candidates for further experimental verification. Domain architecture is displayed in an informative graphic, and detailed results are also available in tabular form. We anticipate that iLIR will assist with elucidating the full complement of LIRCPs in eukaryotes.     

A note on the stability of Rappaport—Vassiliadis enrichment medium

After pre-enrichment in buffered peptone water, 376 samples from chicken carcasses, minced meat, pork sausages, faeces of healthy pigs and sewage-polluted seawater were enriched in Rappaport—Vassiliadis medium prepared either 4 d or 6–7 months before use. It was observed that the two media were equally effective in detecting Salmonella spp., (82 positive samples with each medium) and in their ability to inhibit competing organisms.     

A comparative chemotaxonomic study of eight taxa of the Dipsacaceae family

ABSTRACT

Eight taxa of the Dipsacaceae family were examined for their fatty acid and sterol composition. Separation and identification of the lipid fraction was achieved by gas chromatography and gas chromatography-mass spectrometry. The phenotypic differences among taxa were established by cluster analysis. Correlation coefficients were obtained to investigate numerical relationships among constituents of the fatty acids. The results showed many significant correlations between different constituents, as well as four clusters of taxa using two linkage types of clustering.     

Photomorphogenic reactions in geranium (Pelargonium x hortotum) plants stimulated by brief exposures of ultraviolet-C irradiation

Of all environmental factors that influence plant growth and development, light and other regions of the electromagnetic spectrum are most essential. In the present study, the effects of ultraviolet-C (UV-C) irradiation on two cultivars, Victor and Glacis, of pot grown geranium (Pelargonium x hortotum) plants were evaluated. Two-year experimental data showed that low doses of UV-C (i.e. 0.5–5.0?kJ?m^?2) irradiation led to photomorphogenic changes recorded as increases of fresh and dry weight, number of lateral stems and number of inflorescences formed. Although changes were recorded for both cultivars, responses to UV-C were population dependent with cv. Glacis appearing to be the more responsive one. The number of inflorescences formed on UV-C irradiated cv. Glacis plants was significantly (P?<?0.05) higher to that of the non-irradiated controls throughout the pot trials in 2010 and 2011. For example, in the 2010 pot trial, exposure to 2.5?kJ?m^?2 UV-C resulted in the increase of inflorescences by 75?%. Additionally, the number of lateral stems on UV-C irradiated plants of cvs. Victor and Glacis increased by 122?% and 64?%, respectively. Temperatures and PARs during the 2-year pot trials varied to a considerable level and seem to have affected floral development and growth of both cultivars. However, only in cv. Glacis the effect on floral development was significant as cv. Victor geraniums grown in 2010 and 2011 showed comparable numbers of inflorescences. Our results show that brief exposures of geranium plants to UV-C may facilitate the production of high quality final products in a cost effective and environmentally friendly way.     

Characterization of MCU-Binding Proteins MCUR1 and CCDC90B — Representatives of a Protein Family Conserved in Prokaryotes and Eukaryotic Organelles

1. Download high-res image (208KB)
2. Download full-size image

     

The use of stable isotope-labeled glycerol and oleic acid to differentiate the hepatic functions of DGAT1 and -2

Diacylglycerol acyltransferase (DGAT) catalyzes the final step in triglyceride (TG) synthesis. There are two isoforms, DGAT1 and DGAT2, with distinct protein sequences and potentially different physiological functions. To date, the ability to determine clear functional differences between DGAT1 and DGAT2, especially with respect to hepatic TG synthesis, has been elusive. To dissect the roles of these two key enzymes, we pretreated HepG2 hepatoma cells with (13)C(3)-D(5)-glycerol or (13)C(18)-oleic acid, and profiled the major isotope-labeled TG species by liquid chromatography tandem mass spectrometry. Selective DGAT1 and DGAT2 inhibitors demonstrated that (13)C(3)-D(5)-glycerol-incorporated TG synthesis was mediated by DGAT2, not DGAT1. Conversely, (13)C(18)-oleoyl-incorporated TG synthesis was predominantly mediated by DGAT1. To trace hepatic TG synthesis and VLDL triglyceride (VLDL-TG) secretion in vivo, we administered D(5)-glycerol to mice and measured plasma levels of D(5)-glycerol-incorporated TG. Treatment with an antisense oligonucleotide (ASO) to DGAT2 led to a significant reduction in D(5)-glycerol incorporation into VLDL-TG. In contrast, the DGAT2 ASO had no effect on the incorporation of exogenously administered (13)C(18)-oleic acid into VLDL-TG. Thus, our results indicate that DGAT1 and DGAT2 mediate distinct hepatic functions: DGAT2 is primarily responsible for incorporating endogenously synthesized FAs into TG, whereas DGAT1 plays a greater role in esterifying exogenous FAs to glycerol.