Cav-1 participates in the development of diabetic neuropathy pain through the TLR4 signaling pathway

This study aims to determine whether caveolin-1 (Cav-1) participates in the process of diabetic neuropathic pain by directly regulating the expression of toll-like receptor 4 (TLR4) and the subsequent phosphorylation of N-methyl-D-aspartate receptor 2B subunit (NR2B) in the spinal cord. Male Sprague-Dawley rats (120–150 g) were continuously fed with high-fat and high-sugar diet for 8 weeks, and received a single low-dose of intraperitoneal streptozocin injection in preparation for the type-II diabetes model. Then, these rats were divided into five groups according to the level of blood glucose, and the mechanical withdrawal threshold and thermal withdrawal latency values. The pain thresholds were measured at 3, 7, and 14 days after animal grouping. Then, eight rats were randomly chosen from each group and killed. Lumbar segments 4–6 of the spinal cord were removed for western blot analysis and immunofluorescence assay. Cav-1 was persistently upregulated in the spinal cord after diabetic neuropathic pain in rats. The downregulation of Cav-1 through the subcutaneous injection of Cav-1 inhibitor daidzein ameliorated the pain hypersensitivity and TLR4 expression in the spinal cord in diabetic neuropathic pain (DNP) rats. Furthermore, it was found that Cav-1 directly bound with TLR4, and the subsequent phosphorylation of NR2B in the spinal cord contributed to the modulation of DNP. These findings suggest that Cav-1 plays a vital role in DNP processing at least in part by directly regulating the expression of TLR4, and through the subsequent phosphorylation of NR2B in the spinal cord.     

Effect of production system and pruning on Aphis sambuci dynamics over time and on elderberry yield

In a 2‐year study, elder aphid (Aphis sambuci) dynamics over time and berry yield were evaluated in two production systems (integrated and organic) and in two winter pruning treatments (trees pruned to four and eight scaffolds) in two black elderberry orchards in Hungary. In the organic production system, the first aphid colony was observed 1–2 weeks earlier (late‐March) in both years and locations compared to the integrated programme. The number of aphid colonies then increased until mid‐May in both years, reaching a maximum number of aphid colonies of 11.2 on 100 scaffolds in the integrated production system and of 38.9 in the organic programme. Then, the number of colonies decreased and reached a zero value at mid‐June in the integrated production system and 2 weeks later (early July) in the organic one in both years and locations. First autumn aphid colonies were observed in early September in the integrated production system but 2 weeks earlier (late August) in the organic one in both years and locations. The number of aphid colonies between mid‐April and mid‐June indicated a larger increase on trees pruned to eight scaffolds compared to trees pruned to four scaffolds. Both the total number of aphid colonies and the area under the aphid colony curves (AUACC) were significantly lower (P < 0.001) in the integrated treatments compared with organic ones. Across all treatments, both measures were significantly lower (P < 0.05) on trees pruned to four scaffolds compared with trees pruned to eight scaffolds. However, when the effect of pruning on the number of aphid colonies was analysed separately for integrated and organic plots, pruning caused significant differences in aphid colony numbers and AUACC in the organic plots. Berry yield was significantly higher (P < 0.05) in the integrated treatments compared with the organic ones, but pruning showed no significant effect on yield. Overall, pruning to four scaffolds resulted in a lower aphid colony number in the organic production system compared to the integrated one. Thus, winter pruning may be useful as an aphid control strategy in organic elderberry orchards.     

Genome-Wide Prediction Methods in Highly Diverse and Heterozygous Species: Proof-of-Concept through Simulation in Grapevine

Nowadays, genome-wide association studies (GWAS) and genomic selection (GS) methods which use genome-wide marker data for phenotype prediction are of much potential interest in plant breeding. However, to our knowledge, no studies have been performed yet on the predictive ability of these methods for structured traits when using training populations with high levels of genetic diversity. Such an example of a highly heterozygous, perennial species is grapevine. The present study compares the accuracy of models based on GWAS or GS alone, or in combination, for predicting simple or complex traits, linked or not with population structure. In order to explore the relevance of these methods in this context, we performed simulations using approx 90,000 SNPs on a population of 3,000 individuals structured into three groups and corresponding to published diversity grapevine data. To estimate the parameters of the prediction models, we defined four training populations of 1,000 individuals, corresponding to these three groups and a core collection. Finally, to estimate the accuracy of the models, we also simulated four breeding populations of 200 individuals. Although prediction accuracy was low when breeding populations were too distant from the training populations, high accuracy levels were obtained using the sole core-collection as training population. The highest prediction accuracy was obtained (up to 0.9) using the combined GWAS-GS model. We thus recommend using the combined prediction model and a core-collection as training population for grapevine breeding or for other important economic crops with the same characteristics.     

Lambda phagemids and their transducing properties

Two recombinant lambda DNAs, lambda gt::pMB9 and lambda NM::pBR322, containing, respectively, the pMB9 and pBR322 replicon were constructed and characterized. Both constructs (phagemid DNAs) transfect Escherichia coli cells, producing mature infectious phage progenies. Alternatively, drug-resistant colonies of transductants can be selected upon infection with these phages (phagemid particles) that maintain phagemid DNA in the cell in the form of covalently closed circular plasmids. The efficiency of transduction for nonlysogenic E. coli strains with lambda gt::pMB9 phage producing lambda repressor cIts ranges from 10(-7) to 10(-2) transductant colonies per input phage, depending on the temperature and strain used, while lambda NM::pBR322 phage carrying imm21 transduces with a frequency of up to 1. This means that each lambda NM::pBR322 phagemid particle is capable of establishing itself in the cell as a nonlethal plasmid, permitting formation of a resistant bacterial colony. The maximal level of transduction with lambda gt::pMB9 was obtained when E. coli cells lysogenic for lambda were used. Thus, we believe that the efficiency of transduction is determined by the turn-on of the phage repressor in the transductant. In addition, we have found that all lambda gt::pMB9-containing transductants under certain conditions harbor precisely excised pMB9; excision of pBR322 from lambda NM::pBR322 has not been observed.