Phloem-based resistance to green peach aphid is controlled by Arabidopsis PHYTOALEXIN DEFICIENT4 without its signaling partner ENHANCED DISEASE SUSCEPTIBILITY1

Green peach aphid (GPA) Myzus persicae (Sülzer) is a phloem-feeding insect with an exceptionally wide host range. Previously, it has been shown that Arabidopsis thaliana PHYTOALEXIN DEFICIENT4 (PAD4), which is expressed at elevated levels in response to GPA infestation, is required for resistance to GPA in the Arabidopsis accession Columbia. We demonstrate here that the role of PAD4 in the response to GPA is conserved in Arabidopsis accessions Wassilewskija and Landsberg erecta. Electrical monitoring of aphid feeding behavior revealed that PAD4 modulates a phloem-based defense mechanism against GPA. GPA spends more time actively feeding from the sieve elements of pad4 mutants than from wild-type plants, and less time feeding on transgenic plants in which PAD4 is ectopically expressed. The activity of PAD4 in limiting phloem sap uptake serves as a deterrent in host-plant choice, and restricts aphid population size. In Arabidopsis defense against pathogens, all known PAD4 functions require its signaling and stabilizing partner EDS1 (ENHANCED DISEASE SUSCEPTIBILITY1). Bioassays with eds1 mutants alone or in combination with pad4 and with plants conditionally expressing PAD4 under the control of a dexamethasone-inducible promoter reveal that PAD4-modulated defense against GPA does not involve EDS1. Thus, a PAD4 mode of action that is uncoupled from EDS1 determines the extent of aphid feeding in the phloem.     

Reactive Impurities in Excipients: Profiling,Identification and Mitigation of Drug–Excipient Incompatibility

Reactive impurities in pharmaceutical excipients could cause drug product instability, leading to decreased product performance, loss in potency, and/or formation of potentially toxic degradants. The levels of reactive impurities in excipients may vary between lots and vendors. Screening of excipients for these impurities and a thorough understanding of their potential interaction with drug candidates during early formulation development ensure robust drug product development. In this review paper, excipient impurities are categorized into six major classes, including reducing sugars, aldehydes, peroxides, metals, nitrate/nitrite, and organic acids. The sources of generation, the analytical method for detection, the stability of impurities upon storage and processing, and the potential reactions with drug candidates of these impurities are reviewed. Specific examples of drug–excipient impurity interaction from internal research and literature are provided. Mitigation strategies and corrective measures are also discussed.     

Premature leaf senescence modulated by the Arabidopsis PHYTOALEXIN DEFICIENT4 gene is associated with defense against the phloem-feeding green peach aphid

Aphids, which are phloem-feeding insects, cause extensive loss of plant productivity and are vectors of plant viruses. Aphid feeding causes changes in resource allocation in the host, resulting in an increase in flow of nutrients to the insect-infested tissue. We hypothesized that leaf senescence, which is involved in the programmed degradation of cellular components and the export of nutrients out of the senescing leaf, could be utilized by plants to limit aphid growth. Using Arabidopsis (Arabidopsis thaliana) and green peach aphid (GPA; Myzus persicae Sulzer), we found that GPA feeding induced premature chlorosis and cell death, and increased the expression of SENESCENCE ASSOCIATED GENES (SAGs), all hallmarks of leaf senescence. Hypersenescence was accompanied by enhanced resistance against GPA in the Arabidopsis constitutive expresser of PR genes5 and suppressor of SA insensitivity2 mutant plants. In contrast, resistance against GPA was compromised in the phytoalexin deficient4 (pad4) mutant plant. The PAD4 gene, which is expressed at elevated level in response to GPA feeding, modulates the GPA feeding-induced leaf senescence. In comparison to the wild-type plant, GPA feeding-induced chlorophyll loss, cell death, and SAG expression were delayed in the pad4 mutant. Although PAD4 is associated with camalexin synthesis and salicylic acid (SA) signaling, camalexin and SA signaling are not important for restricting GPA growth; growth of GPA on the camalexin-biosynthesis mutant, pad3, and the SA deficient2 and NahG plants and the SA-signaling mutant, nonexpresser of PR genes1, were comparable to that on the wild-type plant. Our results suggest that PAD4 modulates the activation of senescence in the aphid-infested leaves, which contributes to basal resistance to GPA.     

Catheter lock technique: in vitro efficacy of ethanol for eradication of methicillin-resistant staphylococcal biofilm compared with other agents

Biofilm formation in central venous catheters (CVC) is a prerequisite for catheter-related bloodstream infection (CRBSI). The catheter lock technique has been used to treat biofilm infection, but the ideal agent, concentration and the minimum exposure time necessary to eradicate the biofilms are not clearly known. In this study, biofilm-producing strains of staphylococci were used to find out the minimum biofilm eradication concentration of ethanol compared with three other conventional antibacterial agents. Eight representative methicillin-resistant staphylococci, from colonized CVCs, were studied. The biofilms were exposed to 1, 5 and 10?mg?mL(-1) of gentamicin, ciprofloxacin and vancomycin. The ethanol concentrations used were 20%, 40% and 80%. Biofilms were examined for the presence of live organisms after exposure to these agents from 30?min to 24?h. The three antibiotics were unable to eradicate the biofilms even after 24?h, while ethanol at 40% concentration could do so for all the isolates in 1?h. Our study highlights the efficacy and rationale of using 40% ethanol for a short period as catheter lock solution to eradicate biofilms and thus to prevent CRBSI, instead of using high concentrations of antibiotics for extended periods.