NanoSIMS Analysis of Intravascular Lipolysis and Lipid Movement across Capillaries and into Cardiomyocytes

1. Download high-res image (306KB)
2. Download full-size image

     

Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens

Background

Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection.

Methods

RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006–2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems.

Results

rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%.

Conclusions

Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries.

     

Remapping of the stripe rust resistance gene <Emphasis Type="Italic">Yr10</Emphasis> in common wheat

Key message

Yr10 is an important gene to control wheat stripe rust, and the search for Yr10 needs to be continued.

Abstract

Wheat stripe rust or yellow rust is a devastating fungal disease caused by Puccinia striiformis f. sp. tritici (Pst). Host disease resistance offers a primary source for controlling wheat stripe rust. The stripe rust resistance gene Yr10 confers the race-specific resistance to most tested Pst races in China including CYR29. Early studies proposed that Yr10 was a nucleotide-binding site, leucine-rich repeat gene archived as GenBank accession AF149112 (hereafter designated the Yr10 candidate gene or Yr10 _CG ). In this study, we revealed that 15 Chinese wheat cultivars positive for Yr10 _CG are susceptible to CYR29. We then expressed the Yr10 _CG cDNA in the common wheat ‘Bobwhite’. The Yr10 _CG -cDNA positive transgenic plants were also susceptible to CYR29. Thus, it is highly unlikely that Yr10 _CG corresponds to the Yr10 resistance gene. Using the Yr10 donor ‘Moro’ and the Pst-susceptible wheat ‘Huixianhong’, we generated two F₃ populations that displayed a single Mendelian segregation on the Yr10 gene, and used them to remap the Yr10 gene. Six markers were placed in the Yr10 region, with the Yr10 _CG gene now mapping about 1.2-cM proximal to the Yr10 locus and the Xsdauw79 marker is completely linked to the Yr10 locus. Apparently, the Yr10 gene has not yet been identified. Fine mapping and positional cloning of Yr10 is important for gene pyramiding for stripe rust resistance in wheat.

     

The population status of breeding Twite Linaria flavirostris in the UK in 2013

Capsule: The second national survey of Twite Linaria flavirostris estimated a UK breeding population of 7831 pairs (95% confidence limits: 5829–10?137) in 2013.

Aims: To estimate the breeding population size of Twite in the UK and constituent countries and to calculate change since the 1999 survey.

Methods: Counts of Twite were made on three visits between May and July across a stratified random sample of 1-km squares in England, Scotland and Wales. In Northern Ireland, a complete census was made of the known range and adjacent 1-km squares with suitable habitat. Field surveys involved walking line transects 200 m apart and, in suitable nesting habitat, making 5-minute stops at 100 m intervals to scan and listen for Twite.

Results: The UK population of Twite was estimated at 7831 pairs (95% CL: 5829–10?137). This was 21% lower but not significantly different from the 1999 survey estimate. Scotland held 98% of the UK population (7640, 95% CL: 5629–9954). There were an estimated 164 pairs (95% CL: 76–297) in England, a significant decline of 72% from 1999. Estimated totals for Wales and Northern Ireland were 16 (95% CL: 10–24) and 18 pairs respectively.

Conclusion: The second national survey suggests a moderate decline in the UK Twite population since 1999 but with considerable variation between countries. Further work is required to understand the drivers of population change across breeding populations.     

Proteomic identification of altered protein O-GlcNAcylation in a triple transgenic mouse model of Alzheimer's disease

PET scan analysis demonstrated the early reduction of cerebral glucose metabolism in Alzheimer disease (AD) patients that can make neurons vulnerable to damage via the alteration of the hexosamine biosynthetic pathway (HBP). Defective HBP leads to flawed protein O-GlcNAcylation coupled, by a mutual inverse relationship, with increased protein phosphorylation on Ser/Thr residues. Altered O-GlcNAcylation of Tau and APP have been reported in AD and is closely related with pathology onset and progression. In addition, type 2 diabetes patients show an altered O-GlcNAcylation/phosphorylation that might represent a link between metabolic defects and AD progression. Our study aimed to decipher the specific protein targets of altered O-GlcNAcylation in brain of 12-month-old 3×Tg-AD mice compared with age-matched non-Tg mice. Hence, we analysed the global O-GlcNAc levels, the levels and activity of OGT and OGA, the enzymes controlling its cycling and protein specific O-GlcNAc levels using a bi-dimensional electrophoresis (2DE) approach. Our data demonstrate the alteration of OGT and OGA activation coupled with the decrease of total O-GlcNAcylation levels. Data from proteomics analysis led to the identification of several proteins with reduced O-GlcNAcylation levels, which belong to key pathways involved in the progression of AD such as neuronal structure, protein degradation and glucose metabolism. In parallel, we analysed the O-GlcNAcylation/phosphorylation ratio of IRS1 and AKT, whose alterations may contribute to insulin resistance and reduced glucose uptake. Our findings may contribute to better understand the role of altered protein O-GlcNAcylation profile in AD, by possibly identifying novel mechanisms of disease progression related to glucose hypometabolism.     

Identification of Genes Involved in Flavonoid Biosynthesis of Chinese Narcissus (Narcissus tazetta L. var. chinensis)

Plant Molecular Biology Reporter - Chinese narcissus (Narcissus tazetta L. var. chinensis Roem.) is a popular flower in Asia. However, flower colors are limited with all cultivars having a white...     

Breeding bird species diversity across gradients of land use from forest to agriculture in Europe

Loss, fragmentation and decreasing quality of habitats have been proposed as major threats to biodiversity world‐wide, but relatively little is known about biodiversity responses to multiple pressures, particularly at very large spatial scales. We evaluated the relative contributions of four landscape variables (habitat cover, diversity, fragmentation and productivity) in determining different components of avian diversity across Europe. We sampled breeding birds in multiple 1‐km² landscapes, from high forest cover to intensive agricultural land, in eight countries during 2001?2002. We predicted that the total diversity would peak at intermediate levels of forest cover and fragmentation, and respond positively to increasing habitat diversity and productivity; forest and open‐habitat specialists would show threshold conditions along gradients of forest cover and fragmentation, and respond positively to increasing habitat diversity and productivity; resident species would be more strongly impacted by forest cover and fragmentation than migratory species; and generalists and urban species would show weak responses. Measures of total diversity did not peak at intermediate levels of forest cover or fragmentation. Rarefaction‐standardized species richness decreased marginally and linearly with increasing forest cover and increased non‐linearly with productivity, whereas all measures increased linearly with increasing fragmentation and landscape diversity. Forest and open‐habitat specialists responded approximately linearly to forest cover and also weakly to habitat diversity, fragmentation and productivity. Generalists and urban species responded weakly to the landscape variables, but some groups responded non‐linearly to productivity and marginally to habitat diversity. Resident species were not consistently more sensitive than migratory species to any of the landscape variables. These findings are relevant to landscapes with relatively long histories of human land‐use, and they highlight that habitat loss, fragmentation and habitat‐type diversity must all be considered in land‐use planning and landscape modeling of avian communities.     

Discovery of 2,6-disubstituted pyrazine derivatives as inhibitors of CK2 and PIM kinases

The design and synthesis of a novel series of 2,6-disubstituted pyrazine derivatives as CK2 kinase inhibitors is described. Structure-guided optimization of a 5-substituted-3-thiophene carboxylic acid screening hit (3a) led to the development of a lead compound (12b), which shows inhibition in both enzymatic and cellular assays. Subsequent design and hybridization efforts also led to the unexpected identification of analogs with potent PIM kinase activity (14f).