Nanoparticle-enzyme hybrid systems for nanobiotechnology

Biomolecule-nanoparticle (NP) [or quantum-dot (QD)] hybrid systems combine the recognition and biocatalytic properties of biomolecules with the unique electronic, optical, and catalytic features of NPs and yield composite materials with new functionalities. The biomolecule-NP hybrid systems allow the development of new biosensors, the synthesis of metallic nanowires, and the fabrication of nanostructured patterns of metallic or magnetic NPs on surfaces. These advances in nanobiotechnology are exemplified by the development of amperometric glucose sensors by the electrical contacting of redox enzymes by means of AuNPs, and the design of an optical glucose sensor by the biocatalytic growth of AuNPs. The biocatalytic growth of metallic NPs is used to fabricate Au and Ag nanowires on surfaces. The fluorescence properties of semiconductor QDs are used to develop competitive maltose biosensors and to probe the biocatalytic functions of proteases. Similarly, semiconductor NPs, associated with electrodes, are used to photoactivate bioelectrocatalytic cascades while generating photocurrents.     

Linking traits of foraging animals to spatial patterns of plants: social and solitary ants generate opposing patterns of surviving seeds

Foraging traits of seed predators are expected to impact the spatial structure of plant populations, community dynamics and diversity. Yet, many of the key mechanisms governing distance- or density-dependent seed predation are poorly understood. We designed an extensive set of field experiments to test how seed predation by two harvester ant species interact with seed dispersal in shaping the spatial patterns of surviving seeds. We show that the Janzen–Connell establishment pattern can be generated by central-place foragers even if their focal point is located away from the seed source. Furthermore, we found that differences in the social behaviour of seed predators influence their sensitivity to seed density gradients and yield opposing spatial patterns of surviving seeds. Our results support the predictions of a recent theoretical framework that unifies apparently opposing plant establishment patterns, and suggest that differences in foraging traits among seed predators can drive divergent pathways of plant community dynamics.     

Osmotic Survival of the Entomopathogenic Nematode Steinernema carpocapsae

The effect of different osmolytes on the viability and the effect of osmotic pressure on the induction of a dormant state similar to that caused by a slow desiccation rate were evaluated in the entomopathogenic nematode Steinernema carpocapsae ‘All’. For both experiments, a high-temperature (45°C) assay (HTA) was employed. Exposing fresh infective juveniles to the HTA resulted in a drastic reduction in viability. Using the same assay, the mortality of desiccated nematodes was gradual, showing an enhanced ability to withstand high-temperature conditions. The patterns of decline in viability in the evaporatively dehydrated and the osmotically desiccated nematodes were similar. Most of the salts tested in the screening assay caused high mortality levels among the nematodes within the first 24 h of exposure. In contrast, the nonionic solutes tested did not hamper the viability of the infective juveniles. In these nonionic solutions, all nematodes were completely shrunk after 48 h. Furthermore, 72-h exposure to these solutions resulted in an increase in heat tolerance similar to that of the evaporatively dehydrated nematodes. A substantial increase in heat tolerance was recorded in the treatments with glycerol solutions at concentrations from 2.2 to 3.8 M. A similar effect was obtained by polyethylene glycol (PEG) 300 MW at concentrations ranging from 1.2 to 1.6 M. PEG 600 MW induced enhancement of heat tolerance at a concentration of 0.8 M. A high level of viability was attained among nematodes that were stored for 72 days following a gradual increase in glycerol concentrations. Exposure of these nematodes to 45°C in the HTA resulted in 87.3 ± 4.7 and 49.2 ± 3.9% survival after 4 and 8 h, respectively. Reduction in viability was observed among nematodes that were directly exposed to the glycerol solution over a 19-day storage period. With this treatment, survival levels of 72.7 ± 3.9 and 26.5 ± 4.7% after 4 and 8 h, respectively, were recorded in the HTA. Reduction in viability among nematodes stored in distilled water was noted after 36 days of storage. Evaluation of nematode infectivity by two criteria (insect mortality and invasion rate) indicated that infectivity of nematodes desiccated by gradual osmotic pressure induced by glycerol was similar to that of fresh nematodes after 54 days in storage at 25°C. In comparison, infectivity of nematodes stored in distilled water declined significantly compared to that of fresh nematodes.     

Detection of <Emphasis Type="Italic">Actinobacillus pleuropneumoniae</Emphasis> by PCR on Field Strains from Healthy and Diseased Pigs

We investigated whether primers able to specifically amplify a 0.7-kb DNA fragment from the conserved cpx genes could be applied to analyze A. pleuropneumoniae field isolates. The specific cpx primers were tested on 120 strains of A. pleuropneumoniae and other NAD-dependent field isolates from healthy and diseased animals to analyze A. pleuropneumoniae isolates from pigs in Brazil. We found that PCR and hybridization were able to discriminate between isolates of A. pleuropneumoniae and other bacteria. The 0.7-kb cpx DNA fragments were amplified from all 63 A. pleuropneumoniae isolates from herds with clinical symptoms and were isolated from lesions of acute cases of swine pleuropneumonia, both serotypable and nonserotypable. The PCR was also applied to 57 field isolates obtained from animals of apparently healthy herds, and the amplified cpx product was present in four serotypable and only two out of eleven A. pleuropneumoniae nonserotypable isolates. All nonserotypable A. pleuropneumoniae isolates revealed the apxA amplification pattern compatible with previously known serotypes. Some nonserotypable isolates might represent a population of isolates that originally were serotypable but lost the ability to react with serotype-specific antisera or might belong to novel serotypes. The PCR method applied is highly sensitive for serotypable A. pleuropneumoniae strains and for nonserotypable strains isolated from acute cases of swine pleuropneumoniae in Brazil. Received: 13 June 2002 / Accepted: 5 August 2002     

Octreotide,a Somatostatin Analogue,Fails to Inhibit Hypoxia-induced Retinal Neovascularization in the Neonatal Rat

Objective: Octreotide, a somatostatin analogue, has been shown to prevent angiogenesis in diverse in vitro models. We evaluated its effect on retinal neovascularization in vivo, using a neonatal rat retinopathy model. Methods: We used, on alternating days, hypoxia (10% O₂) and hyperoxia (50% O₂) during the first 14 days of neonatal rats, to induce retinal neovascularization. Half of the rats were injected subcutaneously with octreotide 0.7 μg/g BW twice daily. At day 18 the eyes were evaluated for the presence of epiretinal and vitreal hemorrhage, neovascularization and epiretinal proliferation. Octreotide pharmacokinetics and its effect on serum growth hormone (GH) and insulin-like growth factor I (IGF-I) were examined in 28 rats. Results: Serum octreotide levels were 667 μg/1 two hours after injection, 26.4 μg/1 after nine hours and 3.2 μg/1 after 14 hours. GH levels were decreased by 40% (p = 0.002) two hours after injection but thereafter returned to baseline. IGF-I levels were unchanged two hours after injection and were elevated by 26% 14 hours after injection (p = 0.02). Epiretinal membranes were highly associated with epiretinal hemorrhages (p < 0.001), while retinal neovascularization was notably associated with vitreal hemorrhages (p < 0.001). Conclusions: Twice-daily injections of octreotide failed to produce sustained decrease in serum GH, but produced rebound elevation of serum IGF-I. Accordingly, no statistically significant effect of injections on retinal pathology was noted. This finding, however, does not contradict our assumption that GH suppression may decrease the severity of retinopathy.     

Studying and modelling dynamic biological processes using time-series gene expression data

Biological processes are often dynamic, thus researchers must monitor their activity at multiple time points. The most abundant source of information regarding such dynamic activity is time-series gene expression data. These data are used to identify the complete set of activated genes in a biological process, to infer their rates of change, their order and their causal effects and to model dynamic systems in the cell. In this Review we discuss the basic patterns that have been observed in time-series experiments, how these patterns are combined to form expression programs, and the computational analysis, visualization and integration of these data to infer models of dynamic biological systems.