Transgenic Rice Plants Expressing the Antifungal AFP Protein from Aspergillus Giganteus Show Enhanced Resistance to the Rice Blast Fungus Magnaporthe Grisea

The Aspergillus giganteus antifungal protein (AFP), encoded by the afp gene, has been reported to possess in vitro antifungal activity against various economically important fungal pathogens, including the rice blast fungus Magnaporthe grisea. In this study, transgenic rice ( Oryza sativa ) constitutively expressing the afp gene was generated by Agrobacterium -mediated transformation. Two different DNA constructs containing either the afp cDNA sequence from Aspergillus or a chemically synthesized codon-optimized afp gene were introduced into rice plants. In both cases, the DNA region encoding the signal sequence from the tobacco AP24 gene was N-terminally fused to the coding sequence of the mature AFP protein. Transgenic rice plants showed stable integration and inheritance of the transgene. No effect on plant morphology was observed in the afp -expressing rice lines. The inhibitory activity of protein extracts prepared from leaves of afp plants on the in vitro growth of M. grisea indicated that the AFP protein produced by the trangenic rice plants was biologically active. Several of the T(2) homozygous afp lines were challenged with M. grisea in a detached leaf infection assay. Transformants exhibited resistance to rice blast at various levels. Altogether, the results presented here indicate that AFP can be functionally expressed in rice plants for protection against the rice blast fungus M. grisea.     

Engineering photoassimilate partitioning in tobacco plants improves growth and productivity and provides pathogen resistance

Expression of pathogenesis-related (PR) genes is part of the plant's natural defense response against pathogen attack. To study the in vivo role and function of the maize PRms protein, tobacco plants were transformed with the PRms cDNA under the control of the CaMV35S promoter. Transgenic tobacco plants grow faster and yield more leaf and seed biomass. By using immunoelectron microscopy, we found that PRms is associated with plasmodesmata in leaves of transgenic tobacco plants. Furthermore, we found that activation of sucrose efflux from photosynthetically active leaves and accumulation of higher levels of sucrose in leaf tissues are characteristic features of PRms tobacco plants. This, in turn, results in the constitutive expression of endogenous tobacco PR genes and resistance to phytopathogens. The expression of multiple plant defense genes can then be achieved by using a single transgene. These data provide a new approach for engineering disease-resistant plants while simultaneously improving plant yield and productivity through the modification of photoassimilate partitioning.     

Plant polyphenol antioxidants and oxidative stress

In recent years there has been a remarkable increment in scientific articles dealing with oxidative stress. Several reasons justify this trend: knowledge about reactive oxygen and nitrogen species metabolism; definition of markers for oxidative damage; evidence linking chronic diseases and oxidative stress; identification of flavonoids and other dietary polyphenol antioxidants present in plant foods as bioactive molecules; and data supporting the idea that health benefits associated with fruits, vegetables and red wine in the diet are probably linked to the polyphenol antioxidants they contain. In this review we examine some of the evidence linking chronic diseases and oxidative stress, the distribution and basic structure of plant polyphenol antioxidants, some biological effects of polyphenols, and data related to their bioavailability and the metabolic changes they undergo in the intestinal lumen and after absorption into the organism. Finally, we consider some of the challenges that research in this area currently faces, with particular emphasis on the contributions made at the International Symposium "Biology and Pathology of Free Radicals: Plant and Wine Polyphenol Antioxidants" held July 29-30, 1999, at the Catholic University, Santiago, Chile and collected in this special issue of Biological Research.     

Pooled spike trains of correlated presynaptic inputs as realizations of cluster point processes

The pooled spike trains of correlated presynaptic terminals acting synchronously upon a single neuron are realizations of cluster point processes: the notions of spikes synchronizing in bursts and of points bunching in clusters are conceptually identical. The primary processes constituent specifies the timing of the cluster series; subsidiary processes and poolings specify burst structure and tightness. This representation and the Poisson process representation of independent terminals complete the formal approach to pooled trains. The notions usefulness was illustrated by expressing physiological questions in terms of those constituents, each possessing a clear biological embodiment; constituents provided the control variables in simulations using leaky integrate-and-fire postsynaptic neurons excited by multiple weak terminals. Regular or irregular primary processes and bursts series determined low or high postsynaptic dispersions. When convergent set synchrony increased, its postsynaptic consequences approached those of single powerful synapses; concomitantly, output spike trains approached periodic, quasiperiodic, or aperiodic behaviors. The sequence in which terminals fired within bursts affected the predictee and predictor roles of presynaptic and postsynaptic spikes; when inhibition was added, EPSP and IPSP delays and order were influential (summation was noncommutative). Outputs to different correlations were heterogeneous; heterogeneity was accentuated by conditioning by variables such as DC biases.     

The GABAA receptor complex in the developing chick optic tectum: characterization of benzodiazepine and ionophore-linked convulsant/barbiturate recognition sites

By use of membrane preparations and incubation conditions optimized for each binding site, we have characterized the benzodiazepine and ionophore-linked-convulsant/barbiturate modulatory sites within the chick tectal GABA_A receptor complex. Using [³H]flunitrazepam (FNZ) and [³⁵S]t-butylbicyclophosphorothionate (TBPS), respectively, as specific radioligand probes for the two sites, we have found in each case one single population of high-affinity, saturable, specific binding sites. The apparent dissociation constants (K_d) show no change during tectal development (9 nM for [³H]FNZ, and 25–28 nM for [³⁵S]TBPS) while the respective densities of binding sites at saturation (B_max) experience in both cases a twofold increase between embryonic day 16 and postnatal day 10. Ligand-specific pharmacological profiles and allosteric interactions between the transmitter and modulatory sites appear to be well preserved in the chick tectal membrane preparations employed in this study.     

Effects of stimulus timing on transmitter release and postsynaptic membrane potential at crayfish neuromuscular junctions

Summary Different synaptic terminals of the single excitor axon to the opener muscle of crayfish (Procambarus clarkii) often release transmitter in a very different manner when stimulated with the same equal-interval, doublet, or triplet patterns. Compared to synapses that show little facilitation (low F_e synapses), highly facilitating (high F_e) synapses show greater percentage increases in several measures of synaptic efficacy when stimulated with any of these patterns. Low F_e synapses usually show the greater absolute changes in these measures of synaptic efficacy. Changes in the span and pattern of doublets and triplets can independently affect both pre- and postsynaptic measures of synaptic efficacy at either low F_e or high F_e synapses.Abbreviations EJP excitatory junctional potential - MJP spontaneous miniature EJP - F _e ratio of EJP at 1 Hz to EJP amplitude at 10 Hz - F ₁ zero-time facilitation - A ₂,B ₂,C ₂ doubly corrected EJP amplitude of a particular pulse - average amplitude of doubly corrected EJPs in a train of equal-interval, doublets, and triplets, respectively - A_m, B_m, C_m maximum depolarization reached by a particular EJP - time constant of decay     

Neutrophil Extracellular Traps Induce Organ Damage during Experimental and Clinical Sepsis

Organ dysfunction is a major concern in sepsis pathophysiology and contributes to its high mortality rate. Neutrophil extracellular traps (NETs) have been implicated in endothelial damage and take part in the pathogenesis of organ dysfunction in several conditions. NETs also have an important role in counteracting invading microorganisms during infection. The aim of this study was to evaluate systemic NETs formation, their participation in host bacterial clearance and their contribution to organ dysfunction in sepsis. C57Bl/6 mice were subjected to endotoxic shock or a polymicrobial sepsis model induced by cecal ligation and puncture (CLP). The involvement of cf-DNA/NETs in the physiopathology of sepsis was evaluated through NETs degradation by rhDNase. This treatment was also associated with a broad-spectrum antibiotic treatment (ertapenem) in mice after CLP. CLP or endotoxin administration induced a significant increase in the serum concentrations of NETs. The increase in CLP-induced NETs was sustained over a period of 3 to 24 h after surgery in mice and was not inhibited by the antibiotic treatment. Systemic rhDNase treatment reduced serum NETs and increased the bacterial load in non-antibiotic-treated septic mice. rhDNase plus antibiotics attenuated sepsis-induced organ damage and improved the survival rate. The correlation between the presence of NETs in peripheral blood and organ dysfunction was evaluated in 31 septic patients. Higher cf-DNA concentrations were detected in septic patients in comparison with healthy controls, and levels were correlated with sepsis severity and organ dysfunction. In conclusion, cf-DNA/NETs are formed during sepsis and are associated with sepsis severity. In the experimental setting, the degradation of NETs by rhDNase attenuates organ damage only when combined with antibiotics, confirming that NETs take part in sepsis pathogenesis. Altogether, our results suggest that NETs are important for host bacterial control and are relevant actors in the pathogenesis of sepsis.     

A Bonhoeffer-van der Pol oscillator model of locked and non-locked behaviors of living pacemaker neurons

A recent investigation of the influence of periodic inhibitory trains on a crayfish pacemaker neuron showed not only well-known locked periodic responses but also intermittent, messy, and hopping responses. This communication studies the responses of the Bonhoeffer-van der Pol (BVP) model with self-sustained oscillation when exposed to periodic pulse trains inputs. The analysis is similar to that used in crayfish and reveals interesting features, both comparable and complementary to those seen in the living preparation.Supported by Trent H. Wells Jr. Inc.