Generation of a purely clonal defective interfering influenza virus

Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus‐based antivirals.     

Within-tree spatial variation in the leaf monoterpenes of Sequoia sempervirens

Location within a tree was analyzed as a source of variation in Sequoia sempervirens leaf monoterpenes. No differences were found for quantitative composition or total yield/dry wt among lower, middle and upper canopy positions. The awlshaped, spirally arranged leaves of vigorous upper shoots showed small quantitative compositional differences, but not differences in total yield. The intermediate leaf form of young sprouts had the most different monoterpene quantitative composition and about three times the total yield of the above two leaf forms. Analysis of a clonal ring of 17 adult trees resulted in coefficients of variation similar to those for samples collected from different canopy levels of the same shoot. Results revealed the sources and magnitudes of experimental error in comparative studies of this species' leaf monoterpenes, and did not support the concept that somatic mutation provides an important source of variation in a large, long-lived organism such as coast redwood.     

The Amount of BCL6 in B Cells Shortly after Antigen Engagement Determines Their Representation in Subsequent Germinal Centers

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UDP-Gal: <Emphasis Type="Italic">N</Emphasis>-acetylglucosamine β 1–4 galactosyltransferase expressing live attenuated parasites as vaccine for visceral leishmaniasis

As compared to cutaneous leishmaniasis, vaccination against visceral leishmaniasis (VL) has received limited attention. In this study, we demonstrate for the first time that an UDP-Galactose: N-acetylglucosamine β 1–4 galactosyltransferase (GenBank Accession No. EF159943) expressing attenuated LD clonal population (A-LD) is able to confer protection against the experimental challenge with the virulent LD AG83 parasite. A-LD was also effective in established leishmania infection. The vaccinated animals showed both cell mediated (in vitro T-cell proliferation, and DTH response) and humoral responses (Th1 type). These results demonstrate the potential of the attenuated clones as an immunotherapeutic and immunoprophylactic agent against visceral leishmaniasis.     

Dihydroquercetin accumulation in the medium of some wild carrot culture subclones

Approximately half of the subclones examined from one clone of the wild carrot cell culture WC63-1-9-1 accumulated dihydroquercetin in the culture medium. The amount of dihydroquercetin accumulated in the medium varied with the subclone used, the size of the inoculum, the medium used and the time of sampling.Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday     

Relation of light and nitrogen source to growth, nitrate reductase and glutamine synthetase activity of jack pine seedlings

Two-month-old jack pine ( Pinus banksiana Lamb.) seedlings were placed in a greenhouse where both nitrogen source and light level were varied. After 4 months, whole seedling biomass, leaf biomass and relative growth rate were greatest in seedlings grown with NH⁺₄/NO/NO⁻₃-N and full light (FL) and least in seedlings grown with NO ⁻₃-N and low light (LL). NO ⁻₃-seedlings grown under full light and NH⁺₄/NO⁻₃-seedlings grown under low light were approximately equal. This indicates that the extra carbon costs of assimilating only NO⁻₃-N were similar to the reduction of carbon fixation resulting from a 50% decrease in photon flux density. Percentage and total nitrogen content of needles were greater in seedlings grown under low light independent of nitrogen fertilization. Percentage and total nitrogen content of roots were higher under low light and lower when fertilized with NO⁻₃.
Nitrate reductase (NR) activity was higher in roots than in needles, while glutamine synthetase (GS) activity was higher in needles than in roots. Low light resulted in decreased NR activity (mg N)⁻¹ in needles, but not in roots. However, no nitrate was detected in the needles in any treatment. GS activity, on the other hand, was greater under low light in both needles and roots. GS activity in needles is most likely involved with the reassimilation rather than the initial assimilation of ammonium. Some implications of these shifts in enzymatic activity for ecological phenomena in forests are discussed.     

Purification and characterization of lymphocytic clonal growth factor (LCGF) derived from human-human hybridoma SH-76 cells

Human-human hybridoma SH-76 cells were found to produce a factor that supported the growth of lymphocytic cells at low densities. The factor was purified from serum-free conditioned medium of the hybridoma cells by a successive application of ammonium sulfate precipitation, DEAE-Toyopearl, TSK G3000 SW and DEAE-5PW column chromatograph. The purified factor was a 72K single protein. The factor showed marked growth stimulating effect on lymphocytic cell lines, but had no effect on the growth of human adhesive cancer cell lines. Thus, the factor is a lymphocytic clonal growth factor (LCGF), as found previously in human plasma (Miyata, 1988). The LCGF of SH-76 cells could be produced in growth factor-free RPMI medium and purified easily from the conditioned medium. The factor is inactivated by heating at over 80°C, but is much more stable than the LCGF in human plasma.     

MORPHOLOGICAL VARIABILITY IN POPULATIONS OF CHRYSOPHYCEAN RESTING CYSTS. I. GENETIC (INTERCLONAL) AND ENCYSTMENT TEMPERATURE EFFECTS ON MORPHOLOGY1

Many chrysophycean species produce resting cysts (statospores) with purportedly species-specific morphology. I investigated variation in the cyst morphology of a single species that may result from genetic differences among the vegetative clones involved and from variation in the temperature of the environment during cyst development. Populations of Dinobryon cylindricum Imhof cysts were produced under defined conditions in vitro and then sampled for morphological analysis based on SEM micrographs. Morphological data is presented and then used in a multivariate discriminant analysis to determine the utility of each morphological character in distinguishing the six populations studied. Results suggest that some features of cyst morphology (i.e. cyst diameter) are invariant among the populations, while other features show distinctive variation. The density of spines covering the cyst body as well as the morphology of those spines appear correlated to the specific clones involved, and thus may represent useful phenotypic genetic markers. The length and definition of both the spines and the cyst collar, on the other hand, are markedly influenced by encystment temperature. The implications of these findings for paleoecological studies is discussed.     

Growth response of Bolboschoenus maritimus ssp. maritimus and B. maritimus ssp. compactus to different trophic conditions

Bolboschoenus maritimus (L.) Palla (=Scirpus maritimus L.) forms extensive stands in the littoral zone of small fishponds and as a weed in rice and maize fields. Within the species, two subspecies are distinguished: Bolboschoenus maritimus subsp. maritimus, B. maritimus subsp. compactus. They differ in ecology, especially in their relationships with trophic conditions and salinity of habitats. To determine growth response of these two types to different nutrient levels, we compared their seasonal development under experimental cultivation at four controlled nutrient levels. Some differences between the subspecies were found to be stable, regardless of nutrient level, namely greater amount of smaller underground tubers and more extensive rhizome system in subsp. compactus compared to less numerous larger tubers and simpler rhizome system in subsp. maritimus. In response to trophic conditions,the plants of subsp. compactus were more resistant to the conditions of the highest trophic level than those of subsp. maritimus, which were stressed. This demonstrates better adaptability and spreading ability of B. maritimus subsp. compactus at high trophic levels.