Structural and molecular interactions of CCR5 inhibitors with CCR5

We have characterized the structural and molecular interactions of CC-chemokine receptor 5 (CCR5) with three CCR5 inhibitors active against R5 human immunodeficiency virus type 1 (HIV-1) including the potent in vitro and in vivo CCR5 inhibitor aplaviroc (AVC). The data obtained with saturation binding assays and structural analyses delineated the key interactions responsible for the binding of CCR5 inhibitors with CCR5 and illustrated that their binding site is located in a predominantly lipophilic pocket in the interface of extracellular loops and within the upper transmembrane (TM) domain of CCR5. Mutations in the CCR5 binding sites of AVC decreased gp120 binding to CCR5 and the susceptibility to HIV-1 infection, although mutations in TM4 and TM5 that also decreased gp120 binding and HIV-1 infectivity had less effects on the binding of CC-chemokines, suggesting that CCR5 inhibition targeting appropriate regions might render the inhibition highly HIV-1-specific while preserving the CC chemokine-CCR5 interactions. The present data delineating residue by residue interactions of CCR5 with CCR5 inhibitors should not only help design more potent and more HIV-1-specific CCR5 inhibitors, but also give new insights into the dynamics of CC-chemokine-CCR5 interactions and the mechanisms of CCR5 involvement in the process of cellular entry of HIV-1.     

The contribution of a beneficial insectary plant <Emphasis Type="Italic">Scaevola aemula</Emphasis> to survival and long-term establishment of flightless <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in greenhouses

The fairy fan flower, Scaevola aemula R. Br., is a primary candidate insectary plant for maintaining populations of generalist predators. We conducted release experiments in greenhouses of cultivated eggplants to evaluate the effects of intercropping S. aemula on the establishment of flightless Harmonia axyridis Pallas. Compared with a monoculture of eggplant, all release experiments showed that flightless H. axyridis remained in greater numbers in plots with S. aemula planted alongside eggplant. In the release experiment of flightless H. axyridis larvae, the incidence of aphids in the plot with transplanted S. aemula was suppressed compared with that in the release plot without transplanted S. aemula. In a laboratory experiment, the longevity of flightless H. axyridis adults on blossom stems of S. aemula was greater than when open flowers and buds were removed, suggesting that the insects fed on floral resources such as the pollen of S. aemula. Our findings showed that the floral resources of S. aemula can enhance aphid suppression by improving the establishment of flightless H. axyridis.     

Overexpression of an Arabidopsis thaliana galactinol synthase gene improves drought tolerance in transgenic rice and increased grain yield in the field

Drought stress has often caused significant decreases in crop production which could be associated with global warming. Enhancing drought tolerance without a grain yield penalty has been a great challenge in crop improvement. Here, we report the Arabidopsis thaliana galactinol synthase 2 gene (AtGolS2) was able to confer drought tolerance and increase grain yield in two different rice (Oryza sativa) genotypes under dry field conditions. The developed transgenic lines expressing AtGolS2 under the control of the constitutive maize ubiquitin promoter (Ubi:AtGolS2) also had higher levels of galactinol than the non‐transgenic control. The increased grain yield of the transgenic rice under drought conditions was related to a higher number of panicles, grain fertility and biomass. Extensive confined field trials using Ubi:AtGolS2 transgenic lines in Curinga, tropical japonica and NERICA4, interspecific hybrid across two different seasons and environments revealed the verified lines have the proven field drought tolerance of the Ubi:AtGolS2 transgenic rice. The amended drought tolerance was associated with higher relative water content of leaves, higher photosynthesis activity, lesser reduction in plant growth and faster recovering ability. Collectively, our results provide strong evidence that AtGolS2 is a useful biotechnological tool to reduce grain yield losses in rice beyond genetic differences under field drought stress.     

Novel Strain of Spiroplasma Found in Flower Bugs of the Genus Orius (Hemiptera: Anthocoridae): Transovarial Transmission,Coexistence with Wolbachia and Varied Population Density

Spiroplasma, a group of small, wall-less, helical, and motile bacteria belonging to the Mollicutes, contains species with diverse life histories. To date, all the Spiroplasma strains that are known to be transmitted vertically in arthropod lineages belong to either the Spiroplasma ixodetis group or the Spiroplasma poulsonii group. Here, we found that a unique strain of Spiroplasma vertically transmitted in predatory flower bugs of the genus Orius belongs to the Spiroplasma insolitum group, which is a group of bacteria phylogenetically closely related to S. insolitum derived from the tickseed sunflower, Bidens sp. (Asterales: Asteraceae). The infection frequencies in natural populations were16.0 % in Orius sauteri (n?=?75), 40.5 % in Orius nagaii (n?=?37), and 8.0 % in Orius minutus (n?=?87). Orius strigicollis was not infected with Spiroplasma (n?=?147). In the early stage of oogenesis (i.e., within the germarium), a large number of bacteria with the typical morphology of Spiroplasma existed, keeping a distance from Wolbachia bacteria. The Spiroplasma population seemed to increase during host development but Wolbachia population did not.     

Ultrasound-mediated interferon β gene transfection inhibits growth of malignant melanoma

We investigated the effects of ultrasound-mediated transfection (sonotransfection) of interferon β (IFN-β) gene on melanoma (C32) both in vitro and in vivo. C32 cells were sonotransfected with IFN-β in vitro. Subcutaneous C32 tumors in mice were sonicated weekly immediately after intra-tumor injection with IFN-β genes mixed with microbubbles. Successful sonotransfection with IFN-β gene in vitro was confirmed by ELISA, which resulted in C32 growth inhibition. In vivo, the growth ratio of tumors transfected with IFN-β gene was significantly lower than the other experimental groups. These results may lead to a new method of treatment against melanoma and other hard-to-treat cancers.     

CYP716A subfamily members are multifunctional oxidases in triterpenoid biosynthesis

Triterpenoids are a diverse group of secondary metabolites that are associated with a variety of biological activities. Oleanolic acid, ursolic acid and betulinic acid are common triterpenoids in plants with diverse biological activities, including antifungal, antibacterial, anti-human immunodeficiency virus (HIV) and/or antitumor activities. In the present study, using the gene co-expression analysis tool of Medicago truncatula, we found a strong correlation between CYP716A12 and β-amyrin synthase (bAS), which encodes the enzyme responsible for the initial cyclization of 2,3-oxidosqualene to β-amyrin (the basic structural backbone of most triterpenoid saponins). Through an in vitro assay, we identified CYP716A12 as a β-amyrin 28-oxidase able to modify β-amyrin to oleanolic acid (through erythrodiol and, possibly, oleanolic aldehyde). We also confirmed its activity in vivo, by expressing CYP716A12 in transgenic yeast that endogenously produce β-amyrin. In addition, CYP716A12 was evaluated for its potential α-amyrin- and lupeol-oxidizing activities. Interestingly, CYP716A12 was able to generate ursolic acid (through uvaol and, possibly, ursolic aldehyde) and betulinic acid (through betulin). Hence, CYP716A12 was characterized as a multifunctional enzyme with β-amyrin 28-oxidase, α-amyrin 28-oxidase and lupeol 28-oxidase activities. We also identified homologs of CYP716A12 in grape (CYP716A15 and CYP716A17) that are involved in triterpenoid biosynthesis, which indicates the highly conserved functionality of the CYP716A subfamily among plants. These findings will be useful in the heterologous production of pharmacologically and industrially important triterpenoids, including oleanolic acid, ursolic acid and betulinic acid.