Poly(ethylene oxide)-grafted thermoplastic membranes for use as cellular hybrid bio-artificial organs in the central nervous system

Poly(acrylonitrile-co-vinyl chloride) (PAN/VC) anisotropic membranes were chemically modified with poly(ethylene oxide) (PEO) (5000 and 20,000 g/mol) by one of two aqueous reactions: (a) acid hydrolysis of the nitrile group to a carboxylic acid with which amine-terminated PEO (PEO-NH(2)) reacted or (b) base reduction of the nitrile group to an amine with which PEO-succinimide (PEO-SC) reacted. Approximately 1.3% of the bulk material was modified with PEO-NH(2) whereas 1.8 to 3.5% was modified with PEO-SC as determined by proton nuclear magnetic resonance ((1)H NMR) and attenuated total reflectance Fourier transform infrared (ATR FTIR) spectra. Approximately 50 to 75% less bovine serum albumin (BSA) adsorbed to PEO-grafted single skin fibers than to unmodified PAN/VC. Transport properties of modified and unmodified fibers were compared by passive diffusion, convective nominal molecular weight cutoff, and hydraulic permeability. Neither hydraulic permeability nor nominal molecular weight cutoff of BSA changed appreciably after surface modification with PEO indicating that pore structure was not adversely affected by the chemistry involved in grafting poly(ethylene oxide). However, in the absence of any membrane conditioning, the apparent diffusion of alpha-chymotrypsinogen (24,000 g/mol) was enhanced in PEO-grafted PAN/VC fibers possibly as a result of reduced sorption of the permeating protein. In vivo biocompatibility in the brain tissue of rats was judged by histological assessment of the host's cellular response to fibers implanted for 30 days; biocompatibility of both PAN/VC and PAN/VC-g-PEO was satisfactory but improved slightly with PEO grafting. (c) 1994 John Wiley & Sons, Inc.     

Growth and yield of lentil and wheat inoculated with three Glomus isolates from Saskatchewan soils

The vesicular-arbuscular mycorrhizal fungi (VAMF) Glomus clarum (Nicol. and Schenck) isolate NT4, G. mosseae (Nicol. and Gerd.) Gerd. and Trappe isolate NT6 and G. versiforme (Karst.) Berch isolate NT7 coexist in wheat field soils in Saskatchewan. This study assessed the response of lentil (Lens esculenta L.) and wheat (Triticum aestivum L.) to monospecific and mixed cultures of these VAMF isolates. Seedlings were inoculated with 100 spores of a VAMF isolate, or an equal mixture of spores of two isolates, and grown in a sterile soil mix in a growth chamber. Both crops responded differently to these different VAMF isolates. In the case of lentil, G. clarum NT4 was more effective than G. mosseae NT6 and G. versiforme NT7, and significantly increased (P<0.05) the shoot dry weight (43%) and grain yield (57%) compared with the uninoculated control. There was a significant positive correlation between the percentage of VAMF colonized roots and shoot dry weight (r=0.672^***) and shoot phosphorus concentration (r=0.608^***) of lentil. In the case of wheat, G. clarum NT4 had no effect on shoot dry weight, but produced significant (P<0.08) increases in grain yield (12%) and the phosphorus concentration of the shoot and grain. Although G. clarum NT4 and G. mosseae NT6 both produced similar levels of VAM colonization in wheat, the only response of wheat to isolate NT6 was an increase in plant height at harvest. The efficacy of G. clarum NT4 on both crops appeared to be related to its ability to produce more arbuscular colonization than G. mosseae NT6. Dual inoculation of seedlings with G. clarum NT4 and G. mosseae NT6 resulted in competition between these two isolates. This was evident from a comparison of plant shoot dry weight and grain yield, and VAMF spore production on the two crops inoculated either with isolate NT4 alone or in combination with NT6. G. mosseae NT6 reduced the efficacy of G. clarum NT4 by 16% when dual inoculated on lentil, but had no effect when the host was wheat. Based on spore production, it was found that G. clarum NT4 was more competitive than G. mosseae NT6 when dual inoculated on lentil or wheat. Isolate NT4 produced ca. 2000 and 500 spores/ 100 g substrate, respectively, in the lentil and wheat pots, which was approximately 2–3 times more spores than those produced by isolate NT6 with either crop. When the plants were dual inoculated, there was a 15–19% reduction in spore production by G. clarum NT4 and a 50–70% decrease in spore production by G. mosseae NT6. Our results show that G. clarum NT4 was more competitive and effective in its ability to colonize and increase the growth and yield of lentil and wheat than G. mosseae NT6 or G. versiforme NT7. The relative performance of isolate NT4 with different host plants suggests that this VAMF isolate exhibits a host preference for lentil.     

Monitoring the persistence of Laccaria bicolor as an ectomycorrhizal symbiont of nursery-grown Douglas fir by PCR of the rDNA intergenic spacer

The large-scale inoculation of selected beneficial ectomycorrhizal fungi in forest nurseries has generated renewed interest in the ecology of these symbiotic fungi. However, information on the dissemination and persistence of introduced symbionts is scarce due to the limitation of the current identification methods. To identify ectomycorrhizal fungi on single root tips, we investigated the polymorphism of the PCR-amplified ribosomal DNA intergenic spacer (IGS) from a wide range of ectomycorrhizal fungi. To investigate the reliability of this molecular approach in large-scale surveys, the dissemination and persistence on Douglas fir seedlings of the introduced Laccaria bicolor S238N were assessed in a forest nursery in the Massif Central (France). Several hundred ectomycorrhizas and fruiting bodies were sampled from plots where control and L. bicolor inoculated-Douglas fir seedlings were grown for 1.5 years. PCR typing of mycorrhizas indicated that trees inoculated with L. bicolor S238N remained exclusively colonized by that isolate (or sexually derived isolates) for the entire test period. In contrast, control seedlings were infected by indigenous isolates of Laccaria laccata and Thelephora terrestris. The molecular evidence for the persistence of the introduced mycobiont despite the competition from indigenous isolates of the same species provides further illustration of the potential of exotic species for large-scale microbial application.     

Alginate-chitosan coacervation in production of artificial seeds

Survival of secondary embryoids of winter oilseed rape (Brassica napus ssp. oleifera cv. Primor) has been used as an assay for the development of artificial seeds involving complex coacervation of alginate (polyanion) with chitosan (polycation). Germination frequency of 100% was achieved for encapsulated embryoids when alginate formed the inner matrix and chitosan the outer layer. When the matrix makeup was reversed, there was no germination of embryoids. The artificial seeds produced were hardened in dilute alkaline solutions of NaOH and Ca(OH)(2). An optimum setting time could be selected based on a quantitative measurement of resistance of hardened capsules to compression and the germination frequency of the encapsulated embryoids. (c) 1993 John Wiley & Sons, Inc.     

Protein toxicity to aphids: Anin vitro test onAcyrthosiphon pisum

Recent progress in plant transformation for insect resistance has increased the interest in the potential toxicity of proteins towards insect pests. While studies have been targeted to a large array of insect species, phloem-feeding Homoptera have not been investigated yet. The paper describes a routine test for screening toxicity and growth inhibition of purified proteins in artificial diets onAcyrthosiphon pisum (Harris). Twenty-five commercially available proteins of different classes were tested and compared to some non-protein chemicals (an insecticide, an antibiotic …).A. pisum proved to be very sensitive to all proteases tested and to some venoms with general cytolytic properties. A plant lectin, concanavalin A, displayed significant toxicity and growth inhibition, while various proteins such as a soybean proteinase inhibitor, a chitinase, and bovine serum albumin showed measurable impairments of growth only at higher dose (≥250 μg.ml⁻¹). Some proteins were without short-term effect onA. pisum physiology. The influence of these results on aphid-plant interactions are discussed.

---

Résumé L'effet de protéines alimentaires sur les insectes phloémophages, dont les pucerons, n'a jamais été étudié. Nous proposons ici un test biologique standardisé sur milieu artificiel permettant d'analyser les effets de différentes classes de protéines sur la physiologie d'A. pisum. La validité de ce test est éprouvée (protocole, reproductibilité) et les différentes données récoltées (mortalité et inhibition de croissance) permettent de définir des paramètres toxicologiques tels que concentration létale 50 ou concentration inhibitrice 50. Cette caractérisation toxicologique a été réalisée sur 25 protéines appartenant à des classes différentes, ainsi que plusieurs substances non protéiques utilisées comme témoin de toxicité (insecticide, antibiotique, inhibiteur de synthèse protéique et glucoside phénolique). Les regroupements de protéines par proximités de profils toxicologiques ont été corrélés aux activités biochimiques des différentes protéines. Les implications de ces résultats sur les interactions plante-puceron sont discutées, ainsi que le potentiel d'une stratégie de création de variétés transgéniques résistantes aux pucerons.

     

Inoculation with native soil improves seedling survival and reduces non-native reinvasion in a grassland restoration

Losses of grasslands have been largely attributed to widespread land-use changes, such as conversion to row-crop agriculture. The remaining tallgrass prairie faces further losses due to biological invasions by non-native plant species, often with resultant ecosystem degradation. Of critical concern for conservation, restoration of native grasslands has been met with little success following eradication of non-native plants. In addition to the direct and indirect effects of non-native invasive plants on beneficial soil microbes, management practices targeting invasive species may also negatively affect subsequent restoration efforts. To assess mechanisms limiting germination and survival of native species and to improve native species establishment, we established six replicate plots of each of the following four treatments: (1) inoculated with freshly collected prairie soil with native seeds; (2) inoculated with steam-pasteurized soil with native seeds; (3) noninoculated with native seeds; or (4) noninoculated/nonseeded control. Inoculation with whole soil did not improve seed germination; however, addition of whole soil significantly improved native species survival, compared to pasteurized soil or noninoculated treatments. Inoculation with whole soil significantly decreased reestablishment of non-native invasive Bothriochloa bladhii (Caucasian bluestem); at the end of the growing season, plots receiving whole soil consisted of approximately 30% B. bladhii cover, compared to approximately 80% in plots receiving no soil inoculum. Our results suggest invasion and eradication efforts negatively affect arbuscular mycorrhizal hyphal and spore abundances and soil aggregate stability, and inoculation with locally adapted soil microbial communities can improve metrics of restoration success, including plant species richness and diversity, while decreasing reinvasion by non-native species.     

Noninvasive semen collection from an adult orangutan

Preservation of the genetic diversity of the captive orangutan, especially the wild-caught founders, is critical in maintaining a long-term population in zoological parks. One solution to the problem of maintaining maximum genetic diversity would be to initiate a program of artificial insemination for genetically underrepresented individuals through the banking and interinstitutional use of cryopreserved semen. However, little is known about basic orangutan semen characteristics, and current methodology is inadequate to support such a program. In this paper, we report the results of semen collection from an adult Sumatran orangutan (Pongo pygmaeus abelli), using an artificial vagina without anesthesia or electrical stimulation. A total of 27 ejaculates were evaluated during a 1-year period. The total and liquid volumes of the ejaculates at 1 h following collection were 6.1 ± 0.6 ml and 2.6 ± 0.4 ml, respectively (mean ± SEM). The liquid portion continued to exude semen for 2 h; however, 90% of the motile sperm was exuded within the first 30 min. The total number of sperm in the ejaculate was 164 ± 10⁶ ± 16.5, and the percentage of motile cells was 60 ± 2.7%. We conclude that the artificial vagina provides a promising technique for semen collection in the orangutan, and view these results as an initial step in developing methods for in vitro sperm capacitation, sperm cryopreservation, and artificial insemination. © 1993 Wiley-Liss, Inc.     

青天葵的人工栽培技术研究

本文报道青天葵的栽培研究结果。采用较大的种苗种植可获得较大的种茎;选用大中种茎,适当密植,在60—70%的荫蔽度条件下产量较高;适当推迟收获期,不但产量高,而且形成的球茎较大,有利于青天蔡的繁殖和生长。     

Screening Capsicum germplasm for resistance to tomato spotted wilt virus (TSWV)

Resistance conferred by the Tsw locus from Capsicum chinense against Tomato spotted wilt virus (TSWV) has been widely used in breeding programmes. Nevertheless, this resistance depends on inoculation conditions, and isolates able to overcome it have already been detected. In this work 29 accessions of several Capsicum species have been mechanically inoculated with TSWV to identify new sources of resistance. Five accessions showed variable percentages of resistant plants, two of which did not show local lesions on inoculated leaves, suggesting that the response was not mediated through hypersensitivity. Two of these accessions also had a remarkable reduced viral accumulation compared to susceptible control. ECU‐973., a C. chinense accession, showed the best performance against TSWV, with 100% resistant plants. This response was confirmed after mechanical inoculation with three different TSWV isolates. The resistance was maintained when the accession was inoculated with TSWV using a high pressure of viruliferous thrips. These results open new possibilities in the development of a durable resistance to TSWV in pepper.