A blocking ELISA for the detection of specific antibodies to bovine respiratory syncytial virus

A blocking enzyme-linked immunosorbent assay (ELISA) has been adapted to detect specific antibodies in bovine sera to respiratory syncytial virus using a horseradish peroxidase-labeled monclonal antibody to the fusion protein of the virus. This assay plus an indirect blocking ELISA and indirect ELISA were used to detect antibodies to the bovine respiratory syncytial virus (BRSV) in 159 field-origin bovine sera. Results of these assays were compared with serum antibody titers measured by the serum neutralization (SN) test. Over a 56-day period, the mean neutralization titers and the mean delta absorbance values for the blocking ELISA, on the same sera, showed similar declines. However, the calculated correlation coefficients between mean SN titer and mean absorbance value for the blocking ELISA of the individual sera ranged from -0.2 to -0.5 depending on the source of sera. Similar values were obtained whether using crude or purified viral antigen in the assays. Corresponding calculated correlation coefficients were generally higher for the indirect blocking ELISA or indirect ELISA than for the blocking ELISA. The blocking ELISA was between 70 and 64% as sensitive as the serum neutralization test with a specificity of 100 or 90% using the crude and purified viral antigen, respectively. The indirect blocking ELISA and indirect ELISA had similar calculated sensitivities and specificities. The blocking ELISA was faster to run than either of the other ELISA’s or the neutralization test. Further, nonspecific background absorbance was obviated because the blocking ELISA detects antibodies to 1 specific viral protein, the fusion protein. These studies suggest that the blocking ELISA should be useful as a serological test for BRSV antibodies

Soil microbial legacies influence freeze–thaw responses of soil

Warmer winters with less snowfall are increasing the frequency of soil freeze–thaw cycles across temperate regions. Soil microbial responses to freeze–thaw cycles vary and some of this variation may be explained by microbial conditioning to prior winter conditions, yet such linkages remain largely unexplored. We investigated how differences in temperature history influenced microbial community composition and activity in response to freeze–thaw cycles. We collected soil microbial communities that developed under colder (high elevation) and warmer (low elevation) temperature regimes in spruce-fir forests, then added each of these soil microbial communities to a sterile bulk-soil in a laboratory microcosm experiment. The inoculated high-elevation cold and low-elevation warm microcosms were subjected to diurnal freeze–thaw cycles or constant above-freezing temperature for 9 days. Then, all microcosms were subjected to a 7-day above-freezing recovery period. Overall, we found that the high-elevation cold community had, relative to the low-elevation warm community, a smaller reduction in microbial respiration (CO2 flux) during freeze–thaw cycles. Further, the high-elevation cold community, on average, experienced lower freeze–thaw-induced bacterial mortality than the warm community and may have partly acclimated to freeze–thaw cycles via increased lipid membrane fluidity. Respiration of both microbial communities quickly recovered following the end of the freeze–thaw treatment period and there were no changes in soil extractable carbon or nitrogen. Our results provide evidence that past soil temperature conditions may influence the responses of soil microbial communities to freeze–thaw cycles. The microbial community that developed under a colder temperature regime was more tolerant of freeze–thaw cycles than the community that developed under a warmer temperature regime, although both communities displayed some level of resilience. Taken together, our data suggest that microbial communities conditioned to less extreme winter soil temperatures may be most vulnerable to rapid changes in freeze–thaw regimes as winters warm, but they also may be able to quickly recover if mortality is low

Predictors of Citations in Neurosurgical Research: A 5-year Follow-Up

Introduction Citation rates are an important measure for the impact of publications. This study is the most comprehensive analysis of predictors for scientific neurosurgical research articles. Methods Scientific articles published in 13 neurosurgical journals in 2015 were selected. Data collected included: article subject, level of evidence (LOE), journal impact factor (IF), authorship, contributing centers, and study design. Citation counts were collected for each article in the Web of Science (WoS), Google Scholar (GS), and Scopus 2.5 and 5 years after publication. A generalized linear mixed effects model using the predictors of search engine, LOE, number of centers, number of authors, and IF was constructed to predict total citation count at 5 years. Results 2867 articles generated 39190 citations in WoS, 61682 in GS, and 43481 in Scopus. The median [interquartile range] number of citations per article was 10 [14] in WoS, 15 [20] in GS, and 11 [15] in Scopus. On average, for every 1 citation in WoS, Scopus and GS identified 1.11 and 1.58 citations, respectively. Significant predictors of citation count in all databases 5 years after publication included search engine, LOE, number of centers, number of authors, number of countries, journal IF, and the month of publication (p<0.05). The article subject (tumor, spine, etc.) did not significantly predict citation counts. Conclusions In the most thorough analysis of citation predictors in the neurosurgical literature, search engine, LOE, number of centers, number of authors, number of countries, journal impact factor, and month of publication influenced citations 5 years after publication

Zoledronate Causes a Systemic Shift of Macrophage Polarization towards M1 In Vivo

Background: Immunomodulatory properties of bisphosphonates (BP) are suggested to contribute to the development of medication-associated osteonecrosis of the jaw (MRONJ). Furthermore, bisphosphonate-derived immune modulation might contribute to the anti-metastatic effect observed in breast cancer patients. Macrophages are potential candidates for the mediation of immunomodulatory effects of bisphosphonates. The study aimed to investigate the influence of bisphosphonates alone and in combination with surgical trauma on systemic macrophage polarization (M1 vs. M2) using an in vivo rat model. Methods: A total of 120 animals were divided into four groups. Groups 2 and 4 were treated with 8 × 40 μg/kg body weight of the BP Zoledronate i.p. (week 0–7). Groups 3 and 4 were exposed to surgical trauma (week 8, tooth extraction + tibia fracture), whereas in Group 1 neither medication nor surgical trauma was applied. After 8, 10, 12 and 16 weeks, skin, lung and spleen were immunohistochemically examined for macrophage polarization via expression analysis of CD68, CD163 and iNOS using a tissue microarray (TMA). Results: A significant shift of macrophage polarization towards M1 was observed in skin, spleen and lung tissue of animals, with and without surgical trauma, treated with BP when compared to those without BP application. Surgical trauma did not cause a significant increase towards M1 polarization. Conclusions: BP application leads to a systemic pro-inflammatory situation in vivo, independent of surgical trauma, as evidenced by the shift in macrophage polarization towards M1 in various somatic tissues. This provides a possible explanation for the clinically observed anti-tumor effect of bisphosphonates and might also contribute to pathogenesis of MRONJ

Intranasal Delivery of Caspase-9 Inhibitor Reduces Caspase-6-Dependent Axon/Neuron Loss and Improves Neurological Function after Stroke

Despite extensive research to develop an effective neuroprotective strategy for the treatment of ischemic stroke, therapeutic options remain limited. Although caspase-dependent death is thought to play a prominent role in neuronal injury, direct evidence of active initiator caspases in stroke and the functional relevance of this activity have not previously been shown. Using an unbiased caspase-trapping technique in vivo, we isolated active caspase-9 from ischemic rat brain within 1 h of reperfusion. Pathogenic relevance of active caspase-9 was shown by intranasal delivery of a novel cell membrane-penetrating highly specific inhibitor for active caspase-9 at 4 h postreperfusion (hpr). Caspase-9 inhibition provided neurofunctional protection and established caspase-6 as its downstream target. The temporal and spatial pattern of expression demonstrates that neuronal caspase-9 activity induces caspase-6 activation, mediating axonal loss by 12 hpr followed by neuronal death within 24 hpr. Collectively, these results support selective inhibition of these specific caspases as an effective therapeutic strategy for stroke.C.M.T.wassupported bythe American Heart Association and National Institutes of Health (NIH)GrantsNS035933 and NS43089. G.S.S. and S.J.S. were supported by NIH Grant CA69381. E.S.C. was supported by NIH Grant NS40409.Peer reviewe

Weather and Financial Risk-Taking: Is Happiness the Channel?

Weather variables, and sunshine in particular, are found to be strongly correlated with financial variables. I consider self-reported happiness as a channel through which sunshine affects financial variables. I examine the influence of happiness on risk-taking behavior by instrumenting individual happiness with regional sunshine, and I find that happy people appear to be more risk-averse in financial decisions, and accordingly choose safer investments. Happy people take more time for making decisions and have more self-control. Happy people also expect to live longer and accordingly seem more concerned about the future than the present, and expect less inflation

Metabolic Versatility and Antibacterial Metabolite Biosynthesis Are Distinguishing Genomic Features of the Fire Blight Antagonist Pantoea vagans C9-1

Smits THM, Rezzonico F, Kamber T, et al. Metabolic Versatility and Antibacterial Metabolite Biosynthesis Are Distinguishing Genomic Features of the Fire Blight Antagonist Pantoea vagans C9-1. PLoS ONE. 2011;6(7): e22247.Background: Pantoea vagans is a commercialized biological control agent used against the pome fruit bacterial disease fire blight, caused by Erwinia amylovora. Compared to other biocontrol agents, relatively little is currently known regarding Pantoea genetics. Better understanding of antagonist mechanisms of action and ecological fitness is critical to improving efficacy. Principal Findings: Genome analysis indicated two major factors contribute to biocontrol activity: competition for limiting substrates and antibacterial metabolite production. Pathways for utilization of a broad diversity of sugars and acquisition of iron were identified. Metabolism of sorbitol by P. vagans C9-1 may be a major metabolic feature in biocontrol of fire blight. Biosynthetic genes for the antibacterial peptide pantocin A were found on a chromosomal 28-kb genomic island, and for dapdiamide E on the plasmid pPag2. There was no evidence of potential virulence factors that could enable an animal or phytopathogenic lifestyle and no indication of any genetic-based biosafety risk in the antagonist. Conclusions: Identifying key determinants contributing to disease suppression allows the development of procedures to follow their expression in planta and the genome sequence contributes to rationale risk assessment regarding the use of the biocontrol strain in agricultural systems

Rare and low-frequency coding variants alter human adult height

Height is a highly heritable, classic polygenic trait with ~700 common associated variants identified so far through genome - wide association studies . Here , we report 83 height - associated coding variants with lower minor allele frequenc ies ( range of 0.1 - 4.8% ) and effects of up to 2 16 cm /allele ( e.g. in IHH , STC2 , AR and CRISPLD2 ) , >10 times the average effect of common variants . In functional follow - up studies, rare height - increasing alleles of STC2 (+1 - 2 cm/allele) compromise d proteolytic inhibition of PAPP - A and increased cleavage of IGFBP - 4 in vitro , resulting in higher bioavailability of insulin - like growth factors . The se 83 height - associated variants overlap genes mutated in monogenic growth disorders and highlight new biological candidates ( e.g. ADAMTS3, IL11RA, NOX4 ) and pathways ( e.g . proteoglycan/ glycosaminoglycan synthesis ) involved in growth . Our results demonstrate that sufficiently large sample sizes can uncover rare and low - frequency variants of moderate to large effect associated with polygenic human phenotypes , and that these variants implicate relevant genes and pathways