Platelet-activating factor mediated effects on human neutrophil function are inhibited by pertussis toxin

Pretreatment of human neutrophils with pertussis toxin inhibits platelet-activating factor-mediated chemotaxis, superoxide generation, aggregation, and release of lysozyme. By contrast, superoxide generation observed in the presence of phorbol-12-myristate-13 acetate is unaffected. Our results suggest that a target protein for pertussis toxin, probably a GTP binding protein termed "Ni", is involved in the actions of platelet-activating factor on human neutrophils.     

A gene encoding a protein with seven zinc finger domains acts on the sexual differentiation pathways of Schizosaccharomyces pombe.

Byr3 was selected as a multicopy suppressor of the sporulation defects of diploid Schizosaccharomyces pombe cells that lack ras1. Like cells mutant at byr1 and byr2, two genes that encode putative protein kinases and that in multiple copies are also suppressors of the sporulation defects of ras1 null diploid cells, cells mutant at byr3 are viable but defective in conjugation. Nucleic acid sequence indicates byr3 has the capacity to encode a protein with seven zinc finger binding domains, similar in structure to the cellular nucleic acid binding protein (CNBP), a human protein that was identified on the basis of its ability to bind DNA. Expression of CNBP in yeast can partially suppress conjugation defects of cells lacking byr3.     

Characterisation of <Emphasis Type="Italic">Thinopyrum bessarabicum</Emphasis> chromosomes through genome-wide introgressions into wheat

Key message

Genome-wide introgressions of Thinopyrum bessarabicum into wheat resulted in 12 recombinant lines. Cytological and molecular techniques allowed mapping of 1150 SNP markers across all seven chromosomes of the J genome.

Abstract

Thinopyrum bessarabicum (2n = 2x = 14, JJ) is an important source for new genetic variation for wheat improvement due to its salinity tolerance and disease resistance. Its practical utilisation in wheat improvement can be facilitated through development of genome-wide introgressions leading to a variety of different wheat–Th . bessarabicum translocation lines. In this study, we report the generation of 12 such wheat–Th . bessarabicum recombinant lines, through two different crossing strategies, which were characterized using sequential single colour and multi-colour genomic in situ hybridization (sc-GISH and mc-GISH), multi-colour fluorescent in situ hybridization (mc-FISH) and single nucleotide polymorphic (SNP) DNA markers. We also detected 13 lines containing different Th. bessarabicum chromosome aberrations through sc-GISH. Through a combination of molecular and cytological analysis of all the 25 lines containing Th. bessarabicum recombinants and chromosome aberrations we were able to physically map 1150 SNP markers onto seven Th. bessarabicum J chromosomes which were divided into 36 segmental blocks. Comparative analysis of the physical map of Th. bessarabicum and the wheat genome showed that synteny between the two species is highly conserved at the macro-level and confirmed that Th. bessarabicum contains the 4/5 translocation also present in the A genome of wheat. These wheat–Th . bessarabicum recombinant lines and SNP markers provide a useful genetic resource for wheat improvement with the latter having a wider impact as a tool for detection of introgressions from other Thinopyrum species containing the J or a closely-related genome such as Thinopyrum intermedium (J^rJ^rJ^vsJ^vsStSt) and Thinopyrum elongatum (E^eE^e), respectively.

     

Transient subversion of CD40 ligand function diminishes immune responses to adenovirus vectors in mouse liver and lung tissues.

First-generation adenovirus vectors will have limited application in gene therapy for chronic diseases because of destructive host immune responses. Important immune effectors include CD8+ T cells, which mediate target cell destruction and ablate transgene expression, and B cells, which produce neutralizing antibodies that block effective readministration of vector. Previous studies indicated that activation of CD4+ T cells by virus capsid proteins is necessary for full realization of effector function of CD8+ T cells and B cells. In this paper, we present a strategy for preventing CD4+ T-cell activation by an adenovirus vector delivered to mouse liver and lung tissues which is based on interfering with T-cell priming via CD40 ligand-CD40 interactions. Adenovirus transgene expression was stabilized in mice genetically deficient in CD40 ligand (CD40L), and neutralizing antibody to adenovirus did not develop, allowing efficient readministration of vector. A transient blockade of T-cell activation with an antibody to CD40L infused into the animal at the time of adenovirus vector-mediated gene transfer led to stabilization of transgene expression and diminished production of neutralizing antibody, allowing readministration of vector. In vitro T-cell assays suggested that a block in the primary activation of CD4+ T cells was responsible for the lack of B-cell- and cytotoxic-T-cell-dependent responses. This suggests a strategy for improving the potential of adenovirus vectors based on administration of an antibody to CD40L at the time of vector administration.     

Endophytes of fescue grasses enhance susceptibility ofPopillia japonica larvae to an entomopathogenic nematode

We evaluated tritrophic level interactions among fungal endophytes (Acremonium spp.) of fescue grasses (Festuca spp.), the root-feeding Japanese beetlePopillia japonica Newman larvae, and the entomopathogenic nematodeHeterorhabditis bacteriophora Poinar. Third-instarP. japonica larvae were introduced into pots containing endophyteinfected or endophyte-free plants of tall fescueFestuca arundinacea Schreber (cultivars Kentucky 31 and Georgia Jesup Improved) and the Chewings fescueFestuca rubra commutata Guad. (cultivars F-93 and Jamestown II). After two weeks, the surviving larvae were recovered, and their susceptibility to nematodes was evaluated in sand columns. Endophytes enhanced the rate of nematode-induced mortality in all cultivars except Georgia Jesup Improved, and increased the proportion of dead larvae with nematodes in all cultivars except Jamestown II. Endophytes in the cultivar Kentucky 31 were associated with improved nematode establishment in the larvae. No effect on nematode reproduction was found. Since endophytes produce biologically active alkaloids, we tested the effects of an ergot alkaloid, ergotamine tartrate, on the feeding behavior and weight ofP. japonica larvae in agar medium. The alkaloid caused feeding deterrence, and reduced the consumption of medium by the larvae, resulting in weight loss. These larvae were more susceptible toH. bacteriophora than the untreated larvae. Unfed ‘starved’ larvae were more susceptible to nematodes than those fed on untreated agar. Our results support the hypothesis that endophyte-induced starvation ofP. japonica would reduce larval vigor, and render them more susceptible to entomopathogenic nematodes.     

Physiological methods to study biofilm disinfection

This report reviews the development of a rapidin situ approach to study the physiological responses of bacteria within biofilms to disinfectants. One method utilized direct viable counts (DVC) to assess the disinfection efficacy when thin biofilms were exposed to chlorine or monochloramine. Results obtained using the DVC method were one log higher than plate count (PC) estimates of the surviving population after disinfection. Other methods incorporated the use of fluorogenic stains, a cryotomy technique to yield thin (5-m) sections of biofilm communities and examination by fluorescence microscopy. The fluorogenic stains used in this approach included 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), which indicates cellular electron transport activity and Rhodamine 123, which responds specifically to proton motive force. The use of these stains allowed the microscopic discrimination of physiologically active bacteria as well as heterogeneities of active cells within thicker biofilms. The results of experiments using these techniques with pure culture and binary population biofilms on stainless steel coupons indicated biocidal activity of chlorine-based disinfectants occurred initially at the bulk-fluid interface of the communities and progressed toward the substratum. This approach provided a unique opportunity to describe the spatial response of bacteria within biofilms to antimicrobial agents and address mechanisms explaining their comparative resistance to disinfection in a way that has not been possible using traditional approaches. Results obtained using this alternative approach were also consistently higher than PC data following disinfection. These observations suggest that traditional methods involving biofilm removal and bacterial enumeration by colony formation overestimate biocide efficacy. Hence the alternative approach described here more accurately indicates the ability of bacteria surviving disinfection to recover and grow as well as demonstrate spatial heterogeneities in cellular physiological activities within biofilms.     

Influence of subsoil zinc on dry matter production,seed yield and distribution of zinc in oilseed rape genotypes differing in zinc efficiency

The effects of Zn supply (+Zn: 1 mg kg^–1 soil, -Zn: no Zn added) in subsoil were examined in three genotypes of Brassica napus (Zhongyou 821, Xinza 2, Narendra) and one genotype of Brassica juncea (CSIRO-1) in a glasshouse experiment in pots (100 cm long, 10.5 cm diameter). The topsoil (upper 20 cm soil in pots) was supplied with Zn in all treatments whereas Zn was either supplied or omitted from the subsoil. Supplying Zn to subsoil significantly increased the root growth in the lower zone, markedly decreased the number of aborted and unfilled pods plant^–1 and significantly increased the number of developed pods plant^–1, number of seeds pod^–1, individual seed weight and overall seed yield. Subsoil Zn also significantly increased the Zn concentration and Zn content of seed and improved the ratio of Zn uptake by seed to total Zn uptake by seed and shoots. These effects of subsoil Zn were more pronounced in Zhongyou 821, Xinza 2 and Narendra compared with CSIRO-1. CSIRO-I had 92% Zn efficiency (ratio of -Zn subsoil seed yield to +Zn subsoil seed yield expressed in percentage) compared with 63% for Zhongyou 821. Among the four genotypes, CSIRO-1 had the lowest Zn concentration in roots and shoots but highest Zn concentration and content in seed, suggesting it has a superior Zn transport mechanism from source (roots) to sink (seed). CSIRO-1 also significantly decreased the rhizosphere pH in lower rooting zone (20-93 cm) in -Zn subsoil treatment compared with +Zn treatment.     

Seed zinc content influences early vegetative growth and zinc uptake in oilseed rape (Brassica napus and Brassica juncea) genotypes on zinc-deficient soil

Low-Zn seed (around 80 ng Zn per seed) and high-Zn seed (around 160 ng Zn per seed) of Zhongyou 821 (a traditional Brassica napus genotype from China found to be Zn-inefficient in our previous experiments), Narendra (Zn-efficient B. napus genotype from Australia) and CSIRO-1 (a Zn-efficient B. juncea genotype from Australia) oilseed rape genotypes were sown in pots containing Zn-deficient siliceous sand fertilized with low Zn supply (0.05 mg Zn kg^–1 soil) or high Zn supply (2.0 mg Zn kg^–1 soil) in a controlled environment. After six weeks, plants derived from the high-Zn seed had better seedling vigour, increased root and shoot growth, more leaf area and chlorophyll concentration in fresh leaf, and higher Zn uptake in shoot compared to those from low-Zn seed at low Zn supply; the impact of high-Zn seed was more marked in Zhongyou 821 compared with CSIRO-1 and Narendra. The influence of high-Zn seed was dissipated at high Zn supply. CSIRO-1 was superior in terms of shoot dry matter production and Zn uptake in shoots at low Zn supply. The results demonstrate that although oilseed rape has very small seeds (about 3 mg per seed weight) compared with wheat (30 mg per seed weight), Zn reserves present in this very small seed still have a strong impact on early vegetative growth as well as on Zn uptake of plants in Zn-deficient soils. The results suggest that sowing high-Zn seed coupled with growing Zn-efficient genotypes may help in sustaining the production of oilseed rape in Zn-deficient soils, and this has implications for improved seed technology.