Food For Thought: Short-Term Fasting Upregulates Glucose Transporters in Neurons and Endothelial Cells,But Not in Astrocytes

Neurochemical Research - Our group previously reported that 6-h fasting increased both insulin II mRNA expression and insulin level in rat hypothalamus. Given that insulin effects on central...     

Host choice and human blood index of Anopheles pseudopunctipennis in a village of the Andean valleys of Bolivia

Background

Knockdown resistance (kdr) is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para -type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques.

Methods

Fluorescence-based assays based on 1) TaqMan probes and 2) high resolution melt (HRM) analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR), Heated Oligonucleotide Ligation Assay (HOLA), Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA) and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost), and safety (requirement for hazardous chemicals).

Results

The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions) and the most specific (with the lowest number of incorrect scores). Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS-PCR, SSOP-ELISA, PCR Dot Blot and HOLA was fairly similar with a small number of failures and incorrect scores.

Conclusion

The results of blind genotyping trials of each assay indicate that where maximum sensitivity and specificity are required the TaqMan real-time assay is the preferred method. However, the cost of this assay, particularly in terms of initial capital outlay, is higher than that of some of the other methods. TaqMan assays using a PCR machine and fluorimeter are nearly as sensitive as real-time assays and provide a cost saving in capital expenditure. If price is a primary factor in assay choice then the AS-PCR, SSOP-ELISA, and HOLA are all reasonable alternatives with the SSOP-ELISA approach having the highest throughput.     

Adhesion of the genome-sequenced <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp. <Emphasis Type="Italic">cremoris</Emphasis> IBB477 strain is mediated by specific molecular determinants

Understanding the nature of mucus-microbe interactions will provide important information that can help to elucidate the mechanisms underlying probiotic adhesion. This study focused on the adhesive properties of the Lactococcus lactis subsp. cremoris IBB477 strain, previously shown to persist in the gastrointestinal tract of germ-free rats. The shear flow-induced detachment of L. lactis cells was investigated under laminar flow conditions. Such a dynamic approach demonstrated increased adhesion to bare and mucin-coated polystyrene for IBB477, compared to that observed for the MG1820 control strain. To identify potential genetic determinants giving adhesive properties to IBB477, the improved high-quality draft genome sequence comprising chromosome and five plasmids was obtained and analysed. The number of putative adhesion proteins was determined on the basis of surface/extracellular localisation and/or the presence of adhesion domains. To identify proteins essential for the IBB477 specific adhesion property, nine deletion mutants in chromosomal genes have been constructed and analysed using adhesion tests on bare polystyrene as well as mucin-, fibronectin- or collagen IV-coated polystyrene plates in comparison to the wild-type strain. These experiments demonstrated that gene AJ89_07570 encoding a protein containing DUF285, MucBP and four Big_3 domains is involved in adhesion to bare and mucin-coated polystyrene. To summarise, in the present work, we characterised the adhesion of IBB477 under laminar flow conditions; identified the putative adherence factors present in IBB477, which is the first L. lactis strain exhibiting adhesive and mucoadhesive properties to be sequenced and demonstrated that one of the proteins containing adhesion domains contributes to adhesion.     

Environmental Productivity and Biodiversity Effects on Invertebrate Community Invasibility

Productivity influences the availability of resources for colonizing species. Biodiversity may also influence invasibility of communities because of more complete use of resource types with increasing species richness. We hypothesized that communities with higher environmental productivity and lower species richness should be more invasible by a competitor than those where productivity is low or where richness is high. We experimentally examined the invasion resistance of herbivorous meiofauna of Jamaican rock pools by a competitor crustacean (Ostracoda: Potamocypris sp. (Brady)) by contrasting three levels of nutrient input and four levels of species richness. Although relative abundance (dominance) of the invasive was largely unaffected by resource availability, increasing resources did increase the success rate of establishment. Effects of species richness on dominance were more pronounced with a trend towards the lowest species richness treatment of 2 resident species being more invasible than those with 4, 6, or 7 species. These results can be attributed to a ‘sampling effect associated with the introduction of Alona davidii (Richard) into the higher biodiversity treatments. Alona dominated the communities where it established and precluded dominance by the introduced ostracod. Our experimental study supports the idea that niche availability and community interactions define community invasibility and does not support the application of a neutral community model for local food web management where predictions of exotic species impacts are needed.     

Are our actions aligned With our evidence? The skinny on changing the landscape of obesity

Recent debate about the role of food deserts in the United States (i.e., places that lack access to healthy foods) has prompted discussion on policies being enacted, including efforts that encourage the placement of full‐service supermarkets into food deserts. Other initiatives to address obesogenic neighborhood features include land use zoning and parks renovations. Yet, there is little evidence to demonstrate that such policies effect change. While we suspect most researchers and policymakers would agree that effective neighborhood change could be a powerful tool in combating obesity, we desperately need strong and sound evidence to guide decisions about where and how to invest.     

Assessment of cues potentially mediating host selection of Leptoglossus occidentalis on Pinus contorta

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Rainforest conversion to smallholder cash crops leads to varying declines of beetles (Coleoptera) on Sumatra

Southeast Asian arthropod biodiversity is in rapid decline, but the variability of responses within taxa has received little attention. Using canopy fogging, we collected ~50,000 beetles (Coleoptera) in (1) lowland rainforest, (2) jungle rubber (rubber agroforest), and smallholder monoculture plantations of (3) rubber and (4) oil palm in Sumatra, across two landscapes and seasons. On average, beetle abundance was more than 50%, and biomass over 75%, lower in rubber and oil palm plantations than in rainforest and jungle rubber. This pattern was influenced by landscape and season. Abundance and biomass declines were similar in Chrysomelidae, Elateridae, and Staphylinidae, but differed in Curculionidae, which were most abundant in oil palm due to the introduced oil palm pollinator Elaeidobius kamerunicus. Across beetle families, species richness in monocultures was reduced by at least 70% compared to rainforest, with beetle richness in jungle rubber being similar to rainforest. Community composition in oil palm plantations differed markedly from the other land-use systems for Chrysomelidae and Curculionidae, but less for Elateridae and Staphylinidae. Turnover contributed more to overall beta diversity than nestedness for all families and land-use systems. Likely undersampling of the beetle community in rainforest suggests that declines of beetle density and diversity are much more severe than reported here, especially for beetle families with many concealed species, such as Staphylinidae. This study provides first evidence that negative responses of beetles to tropical land-use change vary among families, and is the first report of its kind from heavily understudied Sumatra.     

A taxonomy of transcriptomic cell types across the isocortex and hippocampal formation

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Human Monoclonal Antibody HCV1 Effectively Prevents and Treats HCV Infection in Chimpanzees

Hepatitis C virus (HCV) infection is a leading cause of liver transplantation and there is an urgent need to develop therapies to reduce rates of HCV infection of transplanted livers. Approved therapeutics for HCV are poorly tolerated and are of limited efficacy in this patient population. Human monoclonal antibody HCV1 recognizes a highly-conserved linear epitope of the HCV E2 envelope glycoprotein (amino acids 412–423) and neutralizes a broad range of HCV genotypes. In a chimpanzee model, a single dose of 250 mg/kg HCV1 delivered 30 minutes prior to infusion with genotype 1a H77 HCV provided complete protection from HCV infection, whereas a dose of 50 mg/kg HCV1 did not protect. In addition, an acutely-infected chimpanzee given 250 mg/kg HCV1 42 days following exposure to virus had a rapid reduction in viral load to below the limit of detection before rebounding 14 days later. The emergent virus displayed an E2 mutation (N415K/D) conferring resistance to HCV1 neutralization. Finally, three chronically HCV-infected chimpanzees were treated with a single dose of 40 mg/kg HCV1 and viral load was reduced to below the limit of detection for 21 days in one chimpanzee with rebounding virus displaying a resistance mutation (N417S). The other two chimpanzees had 0.5–1.0 log₁₀ reductions in viral load without evidence of viral resistance to HCV1. In vitro testing using HCV pseudovirus (HCVpp) demonstrated that the sera from the poorly-responding chimpanzees inhibited the ability of HCV1 to neutralize HCVpp. Measurement of antibody responses in the chronically-infected chimpanzees implicated endogenous antibody to E2 and interference with HCV1 neutralization although other factors may also be responsible. These data suggest that human monoclonal antibody HCV1 may be an effective therapeutic for the prevention of graft infection in HCV-infected patients undergoing liver transplantation.