Pathogen-induced vascular gels: Ethylene as a host intermediate

A cell free culture filtrate from 6-day cultures of Fusarium oxysporum f. sp. cubense was processed to give: (1) a heterogeneous enzyme mixture, (2) purified polygalacturonase (PG), (3) partially-purified polygalacturonate lyase and (4) β-1,4-xylanase. When introduced into explanted castor bean leaves each of these preparations was able to promote the formation of vascular system-obstructing gels. Exposure of castor bean leaves to ethylene (3 ppm) also triggered gel formation. Explanted leaves produced ethylene in response to the enzyme mixture and PG. Vascular gel formation did not occur when ethylene production in response to enzymes was prevented.     

Characterization of a conserved T cell epitope in HIV-1 gp41 recognized by vaccine-induced human cytolytic T cells.

A human CTL epitope located in a region of the HIV-1 envelope protein gp41 that is highly conserved among various HIV-1 strains was identified. This epitope was recognized by CD4+ CTL clones that were induced in seronegative humans by immunization with recombinant gp160. Fusion proteins carrying portions of the HIV-1 env gene and synthetic peptides were used to localize this epitope to amino acids 584-595 of the HIV-1 BRU env sequence. Only two positions within this epitope showed variation among North American HIV-1 isolates, and the substitutions were conservative in nature. The Lys to Arg substitution at position 593 abolished recognition, probably by interfering with the peptide-MHC interactions. This epitope was recognized in association with at least one subtype of the widely distributed human class II MHC specificity DPw4, namely DPw4.2. The relatively high frequency of this allele (27.2% among Caucasians) makes it likely that a larger fraction of the population would generate a response directed at this epitope than would be the case for epitopes recognized in the context of gene products of most other class II and class I loci. Interestingly, the closely related DP beta-chain allele types 4.1 and 2.1, which differ from 4.2 by 3 and 1 amino acids, respectively, were unable to present this gp41 peptide to DPw4.2-restricted clones. Comparison of the structure of this epitope with that of other peptides recognized in the context of DPw4.2 led to the identification of a consensus sequence for DPw4.2 binding peptides. Because the gp41 CTL epitope 584-595 identified here is highly conserved and is recognized in the context of a common DP allele, it may represent an important target region for vaccine development. Our results indicate that vaccines containing this epitope may induce in a significant fraction of those immunized CTL active against at least half of all HIV-1 strains.     

Schistosoma mansoni, S. haematobium, and S. japonicum: identification of genus- and species-specific antigenic egg glycoproteins

Immunoreactive egg glycoproteins of Schistosoma mansoni, S. haematobium, and S. japonicum which are genus- and species-specific, or react with sera of patients infected with other parasites, have been identified. Egg proteins were labeled with Iodine-125, and the concanavalin A-binding glycoproteins were immunoprecipitated with sera of patients infected with one of four species of Schistosoma or Trichinella spiralis, Taenia solium, Echinococcus granulosus, Entamoeba histolytica, or Wuchereria bancrofti. These immunoprecipitates were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Despite the strikingly different patterns of glycoproteins of the African species, the antibody immune responses of patients infected with S. mansoni and S. haematobium were found to be so similar that differentiation could not be established. In contrast, sera of patients infected with S. japonicum, S. mekongi, or parasites not of the genus Schistosoma, immunoprecipitated fewer of the major S. mansoni or S. haematobium glycoproteins. Likewise, antibody immune responses of patients infected with the Oriental schistosomes (S. japonicum and S. mekongi) could not be differentiated. Only a few quantitative differences were noted between our S. mansoni egg glycoprotein extract and a standardized soluble egg antigen extract. This study provides an explanation for the extensive cross-reactivity observed in diagnostic assays which utilize various fractions of schistosomal egg extracts as the antigen.     

A modified matrix perfusion-microcarrier bead cell culture system. I. Adaptation of the matrix perfusion system for growth of human foreskin fibroblasts

Two strains of human foreskin fibroblast cells were incapable of sustained growth in a matrix perfusion culture system, possibly because of their inability to attach to the fiber surfaces. Addition of microcarrier beads to the extracapillary space allowed attaining high cell densities in excess of 10(7) cells per culture unit. Microcarrier beads were tested in hollow fiber culture devices containing membranes of 10(4) or 10(5) D nominal porosities. Best results were obtained when initial cell densities of at least (2-3) x 10(6) cells were used in units with 10(5) D pore size membranes and DEAE-Sephadex or polyacryl-amide microcarrier beads in the extracapillary space. This extension of the matrix perfusion system should be useful for growing other anchorage dependent cells while retaining the advantages of perfusion culture.     

Neonatal neuromuscular parameters vary in susceptibility to postnatal ACTH/MSH 4-10 administration

Neonatal Sprague-Dawley rats administered the fragment of adrenocorticotropic hormone ACTH/MSH 4-10 (10 micrograms/kg/daily, SC) postnatally, show marked differences in the plasticity of the functional and morphological parameters of their neuromuscular system. Initial contraction durations of the immature fast muscle, extensor digitorum longus (EDL), are shorter than saline-treated controls indicating accelerated development. Qualitative studies of the developing EDL neuromuscular junctions as viewed by the scanning electron microscope and quantitative analysis permitted by light microscopy confirms that ACTH/MSH 4-10 affects the maturation of the endplate region. Motor behavior of rat pups demonstrates an age-related difference in the susceptibility to this peptide fragment; one week old neonates showing no response to ACTH/MSH 4-10, two week old pups showing an increase in motor activity. The results indicate that while the developing neuromuscular system is sensitive to the input of ACTH/MSH peptide treatment, this susceptibility is age-related.     

A synthetic peptide with sequence identity to the transmembrane protein GP41 of HIV-1 inhibits distinct lymphocyte activation pathways dependent on protein kinase C and intracellular calcium influx.

A synthetic peptide containing env amino acid (aa) sequence 581 to 597 of the transmembrane protein gp41 of human immunodeficiency virus type 1 (HIV-1) was tested for its effect on protein kinase C (PKC) and cytoplasmic free Ca2+ [( Ca2+]i) influx-dependent immune functions. We have previously shown that this peptide inhibits PKC-mediated phosphorylation and T-cell receptor-mediated [Ca2+]i influx as well as lymphoproliferation. In this study we demonstrate that the HIV-1 gp41 peptide aa581-597 inhibits lymphoproliferation stimulated via the distinct T-cell-activation molecules CD3, CD2, and CD28, as well as direct stimulation mediated by phorbol ester combined with ionomycin. Further, aa581-597 inhibits both PKC-dependent interleukin 2 (IL 2) production and the [Ca2+]i influx-dependent but PKC-independent induction of IL 2 receptor expression. The HIV-1 gp41 peptide also induces dramatic morphologic changes in lymphocytes, characterized by cytoplasmic ballooning and the acquisition of adherence to plastic, and these changes are dependent on both the length and the temperature of exposure. The results of this study suggest that the HIV-1 gp41 sequence aa581-597 acts at multiple sites to inhibit both PKC activity and [Ca2+]i influx, resulting in the abrogation of several distinct immune functions that are critical for an intact immune response and are defective in HIV-1-infected individuals.     

Implementing behaviour in individual-based models using neural networks and genetic algorithms

Even though individual-based models (IBMs) have become very popular in ecology during the last decade, there have been few attempts to implement behavioural aspects in IBMs. This is partly due to lack of appropriate techniques. Behavioural and life history aspects can be implemented in IBMs through adaptive models based on genetic algorithms and neural networks (individual-based-neural network-genetic algorithm, ING). To investigate the precision of the adaptation process, we present three cases where solutions can be found by optimisation. These cases include a state-dependent patch selection problem, a simple game between predators and prey, and a more complex vertical migration scenario for a planktivorous fish. In all cases, the optimal solution is calculated and compared with the solution achieved using ING. The results show that the ING method finds optimal or close to optimal solutions for the problems presented. In addition it has a wider range of potential application areas than conventional techniques in behavioural modelling. Especially the method is well suited for complex problems where other methods fail to provide answers. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inference of biogeographic history by formally integrating distinct lines of evidence: genetic,environmental niche and fossil

A primary focus of historical biogeography is to understand changes in species ranges, abundance and genetic connectivity, and changes in community composition. Traditionally, biogeographic inference has relied on distinct lines of evidence, including DNA sequences, fossils and hindcasted ecological niche models. In this review we propose that the development of integrative modeling approaches that leverage multiple distinct data types from diverse disciplines has the potential to revolutionize the field of biogeography. Although each data type contains information on a distinct aspect of species’ biogeographic histories, few studies formally integrate multiple types in analysis. For example, post hoc congruence among analyses based on different data types (e.g. fossils and genetics) is commonly assumed to indicate likely biogeographic histories. Unfortunately, analyses of different data often reach discordant conclusions. Thus, fundamental and unresolved debates continue regarding speed and timing of postglacial migration, location and size of glacial refugia, and degree of long distance dispersal. Formal statistical integration can help address these issues. More specifically, formal integration can leverage all available evidence, account for inherent biases associated with different data types, and quantify data and process uncertainty. Novel, quantitative integration of data and models across fields is now possible due to recent advances in cyberinfrastructure, spatial modeling, online and aggregated ecological databases, data processing and quantitative methods. Our purpose is to make the case for and give examples of rigorous integration of genetic, fossil and environmental/occurrence data for inferring biogeographic history. In particular, we 1) review the need for such a framework; 2) explain common data types and approaches used to infer biogeographic history (and the challenges with each); 3) review state‐of‐the‐art examples of data integration in biogeography; 4) lay out a series of novel, suggested improvements on current methods; and 5) provide an outlook on technical feasibility and future opportunities.